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Hematopoiesis (blood formation) occurs throughout the lifetime of an organism. Long-term sustained hematopoiesis is enabled by hematopoietic stem cells (HSCs) and high-end progenitors acting at the foundation of a differentiation hierarchy. This HSC-driven hierarchy is known as the definitive (adult) system. Despite their critical involvement in adult hematopoiesis, HSCs are not the first blood cells to form in the embryo. An outstanding point of investigation in the field is how and when the shift from an embryonic hematopoietic system, driven by waves of HSC-independent progenitors, switches to the definitive (HSC-driven) hierarchy. In this thesis, I present my investigations into the molecular mechanisms and functional heterogeneity of HSCs and multipotent hematopoietic progenitor cells (MPPs) during fetal life. Specifically, I have: (1) Investigated the role of ZFP831 (a putative transcription factor) in the regulation of HSCs and MPPs development and differentiation. (2) Used single cell RNA sequencing, cellular barcoding, and transplantation experiments, to investigate the transcriptional character and in vivo differentiation capacity of fetal HSCs and MPPs. In this thesis, I demonstrate that functionally diverse MPP subtypes do exist within the embryo. Contrary to the expectations laid out in the literature, clonal investigation revealed that fetal MPPs are not truly multipotent but are comprised of cells with uni-potent and bi-potent differentiation potential. Most importantly, I have discovered a previously immunophenotypically hidden cohort of long-term reconstituting HSCs. These findings provide a more detailed understanding of the hierarchical organisation of HSC and progenitor cells during fetal life.

The immunological response in contact hypersensitivity is incited by small electrophilic compounds, known as haptens, that react with endogenous proteins after skin absorption. However, the identity of hapten-modified proteins seen as immunogenic remains as yet largely unknown. In a recent study, we have for the first time identified a hapten-modified protein in the local lymph nodes of mice treated topically with the model hapten tetramethylrhodamine isothiocyanate (TRITC). The TRITC modification was located on the N-terminal proline of the protein macrophage migration inhibitory factor (MIF). The focus of the current study was to investigate the presence of the same hapten-protein conjugate in blood samples from mice treated topically with TRITC. Furthermore, TRITC modifications of the two major blood proteins, namely hemoglobin (Hb) and albumin (Alb), as well as TRITC modifications of MIF other than the N-terminal proline, were examined. Following incubation with different molar ratios of TRITC, a proteomic approach was applied to characterize conjugate formation of the three aforementioned proteins, using high resolution mass spectrometry (HRMS). The targeted screening of the TRITC-treated mice blood and lymph node samples for these sites led to the identification of only the same TRITC-MIF conjugate previously detected in the lymph nodes. No Hb and Alb conjugates were detected. Quantification of both the TRITC-modified and unmodified N-terminal peptide of MIF in blood and lymph node samples gave interesting insights of MIF’s role in murine contact hypersensitivity. Incubation of MIF with four different haptens encompassing different reactivity mechanisms and potencies, showed adduct formation at different amino acid residues, suggesting that MIF can be the preferred target for a wide variety of haptens. The present study provides essential progress toward understanding of hapten-protein conjugate formation in contact hypersensitivity and identifies hapten-modif

There is increased concern that the quality, generalizability and reproducibility of biomedical research can be influenced by the sex of animals used. We studied the differences between male and female mice in response to invasive pulmonary mucormycosis including susceptibility to infection, host immune reaction and responses to antifungal therapy. We used diabetic ketoacidotic (DKA) or neutropenic mice infected with either Rhizopus delemar or Mucor circinelloides. The only difference detected was that when DKA mice were infected with M. circinelloides, female mice were more resistant to infection than male mice (median survival time of 5 vs. 2 days for female and male mice, respectively). However, a 100% lethality was detected among infected animals of both sexes. Treatment with either liposomal amphotericin B (L-AMB) or posaconazole (POSA) protected mice from infection and eliminated the difference seen between infected but untreated female and male mice. Treatment with L-AMB consistently outperformed POSA in prolonging survival and reducing tissue fungal burden of DKA and neutropenic mice infected with R. delemar or M. circinelloides, in both mouse sexes. While little difference was detected in cytokine levels among both sexes, mucormycosis infection in the DKA mouse model induced more inflammatory cytokines/chemokines involved in neutrophil (CXCL1) and macrophage (CXCL2) recruitment vs. uninfected mice. As expected, this inflammatory response was reduced in the neutropenic mouse model. Our studies show that there are few differences between female and male DKA or neutropenic mice infected with mucormycosis with no effect on the outcome of treatment or host immune response.

There is increased concern that the quality, generalizability and reproducibility of biomedical research can be influenced by the sex of animals used. We studied the differences between male and female mice in response to invasive pulmonary mucormycosis including susceptibility to infection, host immune reaction and responses to antifungal therapy. We used diabetic ketoacidotic (DKA) or neutropenic mice infected with either Rhizopus delemar or Mucor circinelloides. The only difference detected was that when DKA mice were infected with M. circinelloides, female mice were more resistant to infection than male mice (median survival time of 5 vs. 2 days for female and male mice, respectively). However, a 100% lethality was detected among infected animals of both sexes. Treatment with either liposomal amphotericin B (L-AMB) or posaconazole (POSA) protected mice from infection and eliminated the difference seen between infected but untreated female and male mice. Treatment with L-AMB consistently outperformed POSA in prolonging survival and reducing tissue fungal burden of DKA and neutropenic mice infected with R. delemar or M. circinelloides, in both mouse sexes. While little difference was detected in cytokine levels among both sexes, mucormycosis infection in the DKA mouse model induced more inflammatory cytokines/chemokines involved in neutrophil (CXCL1) and macrophage (CXCL2) recruitment vs. uninfected mice. As expected, this inflammatory response was reduced in the neutropenic mouse model. Our studies show that there are few differences between female and male DKA or neutropenic mice infected with mucormycosis with no effect on the outcome of treatment or host immune response.

Arising from mostly de novo mutations, achondroplasia (ACH) is one of the most common, non-lethal forms of chondrodysplasia. The short stature indicative of ACH stems from a gain of function mutation within the complex FGFR3 signaling pathway—mutations mitigating the too-rapid ossification of cartilage to bone. Meclizine, an FDA-approved drug long prescribed for motion sickness, halts such a conversion and allows the reconstitution of chondrodysplasia cell lines in attempts at following a normal growth pattern. Evinced by various cell line rescues as well as increased long bone growth, it can be hypothesized that maternally administered meclizine can rescue the ACH phenotype enough to attenuate the largest danger to ACH infants, foramen magnum stenosis (FMS).

The brain vasculature has several specific features, one of them being the blood-brain barrier (BBB), which supports and protects the brain by allowing for the passage of oxygen and nutrients, while at the same time preventing passage of pathogens and toxins. The BBB also prevents efficient delivery of drugs to the brain, e.g. for treatment of brain tumors. In the murine brain, perivascular fibroblasts were recently identified as a novel potential constituent of the BBB. Here we present the existence of human cells that could be the equivalent to the murine brain perivascular fibroblasts. Using RNA sequencing, we show a similar transcriptomic profile of cultured human brain cells and murine perivascular fibroblasts. These data open up a window for new hypotheses on cell types involved in human CNS diseases.

The brain vasculature has several specific features, one of them being the blood-brain barrier (BBB), which supports and protects the brain by allowing for the passage of oxygen and nutrients, while at the same time preventing passage of pathogens and toxins. The BBB also prevents efficient delivery of drugs to the brain, e.g. for treatment of brain tumors. In the murine brain, perivascular fibroblasts were recently identified as a novel potential constituent of the BBB. Here we present the existence of human cells that could be the equivalent to the murine brain perivascular fibroblasts. Using RNA sequencing, we show a similar transcriptomic profile of cultured human brain cells and murine perivascular fibroblasts. These data open up a window for new hypotheses on cell types involved in human CNS diseases.

The brain vasculature has several specific features, one of them being the blood-brain barrier (BBB), which supports and protects the brain by allowing for the passage of oxygen and nutrients, while at the same time preventing passage of pathogens and toxins. The BBB also prevents efficient delivery of drugs to the brain, e.g. for treatment of brain tumors. In the murine brain, perivascular fibroblasts were recently identified as a novel potential constituent of the BBB. Here we present the existence of human cells that could be the equivalent to the murine brain perivascular fibroblasts. Using RNA sequencing, we show a similar transcriptomic profile of cultured human brain cells and murine perivascular fibroblasts. These data open up a window for new hypotheses on cell types involved in human CNS diseases.

The brain vasculature has several specific features, one of them being the blood-brain barrier (BBB), which supports and protects the brain by allowing for the passage of oxygen and nutrients, while at the same time preventing passage of pathogens and toxins. The BBB also prevents efficient delivery of drugs to the brain, e.g. for treatment of brain tumors. In the murine brain, perivascular fibroblasts were recently identified as a novel potential constituent of the BBB. Here we present the existence of human cells that could be the equivalent to the murine brain perivascular fibroblasts. Using RNA sequencing, we show a similar transcriptomic profile of cultured human brain cells and murine perivascular fibroblasts. These data open up a window for new hypotheses on cell types involved in human CNS diseases.

Cytomegalovirus (CMV) causes clinically important diseases in immune compromised and immune immature individuals. Based largely on work in the mouse model of murine (M)CMV, there is a consensus that myeloid cells are important for disseminating CMV from the site of infection. In theory, such dissemination should expose CMV to cell-mediated immunity and thus necessitate evasion of T cells and NK cells. However, this hypothesis remains untested. We constructed a recombinant MCMV encoding target sites for the hematopoietic specific miRNA miR-142-3p in the essential viral gene IE3. This virus disseminated poorly to the salivary gland following intranasal or footpad infections but not following intraperitoneal infection in C57BL/6 mice, demonstrating that dissemination by hematopoietic cells is essential for specific routes of infection. Remarkably, depletion of NK cells or T cells restored dissemination of this virus in C57BL/6 mice after intranasal infection, while dissemination occurred normally in BALB/c mice, which lack strong NK cell control of MCMV. These data show that cell-mediated immunity is responsible for restricting MCMV to hematopoietic cell-mediated dissemination. Infected hematopoietic cells avoided cell-mediated immunity via three immune evasion genes that modulate class I MHC and NKG2D ligands (m04, m06 and m152). MCMV lacking these 3 genes spread poorly to the salivary gland unless NK cells were depleted, but also failed to replicate persistently in either the nasal mucosa or salivary gland unless CD8+ T cells were depleted. Surprisingly, CD8+ T cells primed after intranasal infection required CD4+ T cell help to expand and become functional. Together, our data suggest that MCMV can use both hematopoietic cell-dependent and -independent means of dissemination after intranasal infection and that cell mediated immune responses restrict dissemination to infected hematopoietic cells, which are protected from NK cells during dissemination by viral immune e

The brain vasculature has several specific features, one of them being the blood-brain barrier (BBB), which supports and protects the brain by allowing for the passage of oxygen and nutrients, while at the same time preventing passage of pathogens and toxins. The BBB also prevents efficient delivery of drugs to the brain, e.g. for treatment of brain tumors. In the murine brain, perivascular fibroblasts were recently identified as a novel potential constituent of the BBB. Here we present the existence of human cells that could be the equivalent to the murine brain perivascular fibroblasts. Using RNA sequencing, we show a similar transcriptomic profile of cultured human brain cells and murine perivascular fibroblasts. These data open up a window for new hypotheses on cell types involved in human CNS diseases.

Aging is characterized by several physiological changes. During the lifespan, the biological systems from the body of humans and other animals remain dynamic. Throughout the early stages of life, the microbiome develops into a complex ecosystem with thousands of species. Variations related to diet, environmental changes, medications affect the diversity and composition of the microbiota through the lifespan. Some old individuals with higher incidence of chronic diseases have a loss of the stability of the microbiome and an imbalance occurs between the different colonizers of the gut, also named dysbiosis. One of the most distinctive changes occurring with age is the prevalent low grade inflammation, which is named inflamm-aging. This not only changes the microbial composition of the GIT but also affects the permeability. Murine models are well established and help us to understand the complex dynamics between the host and the microbial communities inhabiting the gastrointestinal tract. These models allow us to analyze microbial communities from tissue and mucosa, from all sections of the gut, which is limited in humans. Methods standardization is an important topic in microbiome research. In chapter 2 it was compared the efficiency of two sample methods, cotton swab and tissue biopsy, in characterizing the mouth microbiota. In recent years, the mouth microbiome is being seen as a diagnostic tool for not only oral diseases but also systemic diseases. As physiological changes occur with aging, the microbiome from the mouth is affected and there is an increase of pathogens present in the oral surfaces. In murine models, cotton swab is a common tool used for sampling the microbiome of the oral cavity. In our study, we observed similar microbial community structure using both methodologies. However, the species Streptococcus danieliae, Moraxella osloensis, and some unclassified members of Streptococcus were affected by the different sampling procedures. In this trial, we, Das Altern ist durch verschiedene physiologische Veränderungen gekennzeichnet. Während der Lebensdauer von Menschen und anderen Tieren bleiben die biologischen Systeme des Körpers dynamisch. In den frühen Lebensphasen entwickelt sich das Mikrobiom zu einem komplexen Ökosystem mit Tausenden von Arten. Variationen in Bezug auf Ernährung, Umweltveränderungen und Medikamenten beeinflussen die Vielfalt und Zusammensetzung der Mikrobiota während des gesamten Lebens. Bei einigen alten Personen mit einer höheren Inzidenz chronischer Krankheiten ist ein Verlust der Stabilität des Mikrobioms zu beobachten und es tritt ein Ungleichgewicht zwischen den verschiedenen Darmbesiedlern auf, was auch als Dysbiose bezeichnet wird. Eine der auffälligsten Veränderungen ist eine im Alter auftretende niedriggradige Entzündung, die als inflamm-aging bezeichnet wird. Diese verändert nicht nur die mikrobielle Zusammensetzung des GIT, sondern beeinflusst auch dessen Permeabilität. Mausmodelle sind gut etabliert und helfen uns, die komplexe Dynamik zwischen dem Wirt und den im Magen-Darm-Trakt lebenden mikrobiellen Gemeinschaften zu verstehen. Diese Modelle erlauben es uns, die mikrobiellen Gemeinschaften in Gewebe und Mucosa in allen Abschnitten des Darms zu analysieren, was im Menschen limitiert ist. Ein wichtiges Thema in der Mikrobiomforschung ist die Standardisierung von Methoden. In Kapitel 2 wurde die Effizienz von den zwei Probenmethoden, Wattestäbchen und Gewebebiopsie, bei der Charakterisierung der Mundmikrobiota verglichen. In den letzten Jahren wurde das Mundmikrobiom als diagnostisches Instrument nicht nur für orale Erkrankungen, sondern auch für systemische Erkrankungen eingesetzt. Mit zunehmendem Alter treten physiologische Veränderungen auf, welche das Mikrobiom im Mund beeinflusst und eine Zunahme von Krankheitserregern auf den Mundoberflächen beobachten lässt. In Mausmodellen sind Wattestäbchen ein übliches Werkzeug zur Probeentnahme des Mikrobioms der Mundhöhle. In unserer S

Aging is characterized by several physiological changes. During the lifespan, the biological systems from the body of humans and other animals remain dynamic. Throughout the early stages of life, the microbiome develops into a complex ecosystem with thousands of species. Variations related to diet, environmental changes, medications affect the diversity and composition of the microbiota through the lifespan. Some old individuals with higher incidence of chronic diseases have a loss of the stability of the microbiome and an imbalance occurs between the different colonizers of the gut, also named dysbiosis. One of the most distinctive changes occurring with age is the prevalent low grade inflammation, which is named inflamm-aging. This not only changes the microbial composition of the GIT but also affects the permeability. Murine models are well established and help us to understand the complex dynamics between the host and the microbial communities inhabiting the gastrointestinal tract. These models allow us to analyze microbial communities from tissue and mucosa, from all sections of the gut, which is limited in humans. Methods standardization is an important topic in microbiome research. In chapter 2 it was compared the efficiency of two sample methods, cotton swab and tissue biopsy, in characterizing the mouth microbiota. In recent years, the mouth microbiome is being seen as a diagnostic tool for not only oral diseases but also systemic diseases. As physiological changes occur with aging, the microbiome from the mouth is affected and there is an increase of pathogens present in the oral surfaces. In murine models, cotton swab is a common tool used for sampling the microbiome of the oral cavity. In our study, we observed similar microbial community structure using both methodologies. However, the species Streptococcus danieliae, Moraxella osloensis, and some unclassified members of Streptococcus were affected by the different sampling procedures. In this trial, we, Das Altern ist durch verschiedene physiologische Veränderungen gekennzeichnet. Während der Lebensdauer von Menschen und anderen Tieren bleiben die biologischen Systeme des Körpers dynamisch. In den frühen Lebensphasen entwickelt sich das Mikrobiom zu einem komplexen Ökosystem mit Tausenden von Arten. Variationen in Bezug auf Ernährung, Umweltveränderungen und Medikamenten beeinflussen die Vielfalt und Zusammensetzung der Mikrobiota während des gesamten Lebens. Bei einigen alten Personen mit einer höheren Inzidenz chronischer Krankheiten ist ein Verlust der Stabilität des Mikrobioms zu beobachten und es tritt ein Ungleichgewicht zwischen den verschiedenen Darmbesiedlern auf, was auch als Dysbiose bezeichnet wird. Eine der auffälligsten Veränderungen ist eine im Alter auftretende niedriggradige Entzündung, die als inflamm-aging bezeichnet wird. Diese verändert nicht nur die mikrobielle Zusammensetzung des GIT, sondern beeinflusst auch dessen Permeabilität. Mausmodelle sind gut etabliert und helfen uns, die komplexe Dynamik zwischen dem Wirt und den im Magen-Darm-Trakt lebenden mikrobiellen Gemeinschaften zu verstehen. Diese Modelle erlauben es uns, die mikrobiellen Gemeinschaften in Gewebe und Mucosa in allen Abschnitten des Darms zu analysieren, was im Menschen limitiert ist. Ein wichtiges Thema in der Mikrobiomforschung ist die Standardisierung von Methoden. In Kapitel 2 wurde die Effizienz von den zwei Probenmethoden, Wattestäbchen und Gewebebiopsie, bei der Charakterisierung der Mundmikrobiota verglichen. In den letzten Jahren wurde das Mundmikrobiom als diagnostisches Instrument nicht nur für orale Erkrankungen, sondern auch für systemische Erkrankungen eingesetzt. Mit zunehmendem Alter treten physiologische Veränderungen auf, welche das Mikrobiom im Mund beeinflusst und eine Zunahme von Krankheitserregern auf den Mundoberflächen beobachten lässt. In Mausmodellen sind Wattestäbchen ein übliches Werkzeug zur Probeentnahme des Mikrobioms der Mundhöhle. In unserer S

The development of a murine model of osmotic demyelinating syndrome (ODS) allowed to study changes incurred in extrapontine zones of the CNS and featured neuron and glial cell changes in the relay thalamic ventral posterolateral (VPL) and ventral posteromedial (VPM) nuclei before, during and after ODS induction, and characterized without immune response. There, the neuron Wallerian-type deteriorations were verified with fine structure modifications of the neuron cell body, including some nucleus topology and its nucleolus changes. Morphologic analyses showed a transient stoppage of transcriptional activities while myelinated axons in the surrounding neuropil incurred diverse damages, previously reported. Even though the regional thalamus myelin deterioration was clearly recognized with light microscopy 248 h after osmotic recovery of ODS, ultrastructure analyses demonstrated that, at that time, the same damaged parenchyma regions contained nerve cell bodies that have already reactivated nucleus transcriptions and neuroplasm translations because peculiar accumulations of fibro-granular materials, similar to those detected in restored ODS astrocytes, were revealed in these restructuring nerve cell bodies. Their aspects suggested to be accumulations of ribonucleoproteins. The findings suggested that progressive neural function's recovery in the murine model could imitate some aspects of human ODS recovery cases., SCOPUS: ar.j, info:eu-repo/semantics/published

Aging is characterized by several physiological changes. During the lifespan, the biological systems from the body of humans and other animals remain dynamic. Throughout the early stages of life, the microbiome develops into a complex ecosystem with thousands of species. Variations related to diet, environmental changes, medications affect the diversity and composition of the microbiota through the lifespan. Some old individuals with higher incidence of chronic diseases have a loss of the stability of the microbiome and an imbalance occurs between the different colonizers of the gut, also named dysbiosis. One of the most distinctive changes occurring with age is the prevalent low grade inflammation, which is named inflamm-aging. This not only changes the microbial composition of the GIT but also affects the permeability. Murine models are well established and help us to understand the complex dynamics between the host and the microbial communities inhabiting the gastrointestinal tract. These models allow us to analyze microbial communities from tissue and mucosa, from all sections of the gut, which is limited in humans. Methods standardization is an important topic in microbiome research. In chapter 2 it was compared the efficiency of two sample methods, cotton swab and tissue biopsy, in characterizing the mouth microbiota. In recent years, the mouth microbiome is being seen as a diagnostic tool for not only oral diseases but also systemic diseases. As physiological changes occur with aging, the microbiome from the mouth is affected and there is an increase of pathogens present in the oral surfaces. In murine models, cotton swab is a common tool used for sampling the microbiome of the oral cavity. In our study, we observed similar microbial community structure using both methodologies. However, the species Streptococcus danieliae, Moraxella osloensis, and some unclassified members of Streptococcus were affected by the different sampling procedures. In this trial, we, Das Altern ist durch verschiedene physiologische Veränderungen gekennzeichnet. Während der Lebensdauer von Menschen und anderen Tieren bleiben die biologischen Systeme des Körpers dynamisch. In den frühen Lebensphasen entwickelt sich das Mikrobiom zu einem komplexen Ökosystem mit Tausenden von Arten. Variationen in Bezug auf Ernährung, Umweltveränderungen und Medikamenten beeinflussen die Vielfalt und Zusammensetzung der Mikrobiota während des gesamten Lebens. Bei einigen alten Personen mit einer höheren Inzidenz chronischer Krankheiten ist ein Verlust der Stabilität des Mikrobioms zu beobachten und es tritt ein Ungleichgewicht zwischen den verschiedenen Darmbesiedlern auf, was auch als Dysbiose bezeichnet wird. Eine der auffälligsten Veränderungen ist eine im Alter auftretende niedriggradige Entzündung, die als inflamm-aging bezeichnet wird. Diese verändert nicht nur die mikrobielle Zusammensetzung des GIT, sondern beeinflusst auch dessen Permeabilität. Mausmodelle sind gut etabliert und helfen uns, die komplexe Dynamik zwischen dem Wirt und den im Magen-Darm-Trakt lebenden mikrobiellen Gemeinschaften zu verstehen. Diese Modelle erlauben es uns, die mikrobiellen Gemeinschaften in Gewebe und Mucosa in allen Abschnitten des Darms zu analysieren, was im Menschen limitiert ist. Ein wichtiges Thema in der Mikrobiomforschung ist die Standardisierung von Methoden. In Kapitel 2 wurde die Effizienz von den zwei Probenmethoden, Wattestäbchen und Gewebebiopsie, bei der Charakterisierung der Mundmikrobiota verglichen. In den letzten Jahren wurde das Mundmikrobiom als diagnostisches Instrument nicht nur für orale Erkrankungen, sondern auch für systemische Erkrankungen eingesetzt. Mit zunehmendem Alter treten physiologische Veränderungen auf, welche das Mikrobiom im Mund beeinflusst und eine Zunahme von Krankheitserregern auf den Mundoberflächen beobachten lässt. In Mausmodellen sind Wattestäbchen ein übliches Werkzeug zur Probeentnahme des Mikrobioms der Mundhöhle. In unserer S

T-cell acute lymphoblastic leukemia (T-ALL) and T-cell acute lymphoblastic lymphoma (T-LBL) are aggressive hematological malignancies that are currently treated with high-dose chemotherapy. Over the last several years, the search toward novel and less-toxic therapeutic strategies for T-ALL/T-LBL patients has largely focused on the identification of cell-intrinsic properties of the tumor cell. However, non-cell-autonomous activation of specific oncogenic pathways might also offer opportunities that could be exploited at the therapeutic level. In line with this, we here show that endogenous interleukin 7 (IL7) can increase the expression of the oncogenic kinase proviral integration site for Moloney-murine leukemia 1 (PIM1) in CD127+ T-ALL/T-LBL, thereby rendering these tumor cells sensitive to in vivo PIM inhibition. In addition, using different CD127+ T-ALL/T-LBL xenograft models, we also reveal that residual tumor cells, which remain present after short-term in vivo chemotherapy, display consistent upregulation of PIM1 as compared with bulk nontreated tumor cells. Notably, this effect was transient as increased PIM1 levels were not observed in reestablished disease after abrogation of the initial chemotherapy. Furthermore, we uncover that this phenomenon is, at least in part, mediated by the ability of glucocorticoids to cause transcriptional upregulation of IL7RA in T-ALL/T-LBL patient-derived xenograft (PDX) cells, ultimately resulting in non-cell-autonomous PIM1 upregulation by endogenous IL7. Finally, we confirm in vivo that chemotherapy in combination with a pan-PIM inhibitor can improve leukemia survival in a PDX model of CD127+ T-ALL. Altogether, our work reveals that IL7 and glucocorticoids coordinately drive aberrant activation of PIM1 and suggests that IL7-responsive CD127+ T-ALL and T-LBL patients could benefit from PIM inhibition during induction chemotherapy.

Galactomannan (GM) detection in biological samples has been shown to predict therapeutic response by azoles and polyenes. In a murine invasive pulmonary aspergillosis model, fosmanogepix or posaconazole treatment resulted in an ∼6- to 7-log reduction in conidial equivalents (CE)/g lung tissue after 96 h versus placebo. Changes in GM levels in BAL fluid and serum mirrored reductions in lung CE, with significant decreases seen after 96 h or 72 h for fosmanogepix or posaconazole, respectively (P < 0.02).

Heart failure continues to be a common and deadly sequela of myocardial infarction (MI). Despite strong evidence suggesting the importance of myocardial mechanics in cardiac remodelling, many MI studies still rely on two-dimensional analyses to estimate global left ventricular (LV) function. Here, we integrated four-dimensional ultrasound with three-dimensional strain mapping to longitudinally characterize LV mechanics within and around infarcts in order to study the post-MI remodelling process. To induce infarcts with varying severities, we separated 15 mice into three equal-sized groups: (i) sham, (ii) 30 min ischaemia-reperfusion, and (iii) permanent ligation of the left coronary artery. Four-dimensional ultrasound from a high-frequency small animal system was used to monitor changes in LV geometry, function and strain over 28 days. We reconstructed three-dimensional myocardial strain maps and showed that strain profiles at the infarct border followed a sigmoidal behaviour. We also identified that mice with mild remodelling had significantly higher strains in the infarcted myocardium than those with severe injury. Finally, we developed a new approach to non-invasively estimate infarct size from strain maps, which correlated well with histological results. Taken together, the presented work provides a thorough approach to quantify regional strain, an important component when assessing post-MI remodelling.

Retroviral replicating vectors (RRVs) have achieved efficient tumor transduction and enhanced therapeutic benefit in a wide variety of cancer models. Here, we evaluated two different RRVs derived from amphotropic murine leukemia virus (AMLV) and gibbon ape leukemia virus (GALV), which utilize different cellular receptors (PiT-2 and PiT-1, respectively) for viral entry, in human osteosarcoma cells. Quantitative RT-PCR showed that low levels of expression of both receptors were observed in normal and non-malignant cells. However, high PiT-2 (for AMLV) and low PiT-1 (for GALV) expression was observed in most osteosarcoma cell lines. Accordingly, AMLV expressing the green fluorescent protein gene infected and replicated more efficiently than GALV in most osteosarcoma cell lines. Furthermore, RRVs expressing the cytosine deaminase prodrug activator gene showed differential cytotoxicity that correlated with the results of viral spread. AMLV-RRV-mediated prodrug activator gene therapy achieved significant inhibition of subcutaneous MG-63 tumor growth over GALV in nude mice. These data indicate that AMLV vectors predominate over GALV in human osteosarcoma cells. Moreover, our findings support the potential utility of the two RRVs in personalized cancer virotherapy on the basis of receptor expression.

Galactomannan (GM) detection in biological samples has been shown to predict therapeutic response by azoles and polyenes. In a murine invasive pulmonary aspergillosis model, fosmanogepix or posaconazole treatment resulted in an ∼6- to 7-log reduction in conidial equivalents (CE)/g lung tissue after 96 h versus placebo. Changes in GM levels in BAL fluid and serum mirrored reductions in lung CE, with significant decreases seen after 96 h or 72 h for fosmanogepix or posaconazole, respectively (P < 0.02).

Heart failure continues to be a common and deadly sequela of myocardial infarction (MI). Despite strong evidence suggesting the importance of myocardial mechanics in cardiac remodelling, many MI studies still rely on two-dimensional analyses to estimate global left ventricular (LV) function. Here, we integrated four-dimensional ultrasound with three-dimensional strain mapping to longitudinally characterize LV mechanics within and around infarcts in order to study the post-MI remodelling process. To induce infarcts with varying severities, we separated 15 mice into three equal-sized groups: (i) sham, (ii) 30 min ischaemia-reperfusion, and (iii) permanent ligation of the left coronary artery. Four-dimensional ultrasound from a high-frequency small animal system was used to monitor changes in LV geometry, function and strain over 28 days. We reconstructed three-dimensional myocardial strain maps and showed that strain profiles at the infarct border followed a sigmoidal behaviour. We also identified that mice with mild remodelling had significantly higher strains in the infarcted myocardium than those with severe injury. Finally, we developed a new approach to non-invasively estimate infarct size from strain maps, which correlated well with histological results. Taken together, the presented work provides a thorough approach to quantify regional strain, an important component when assessing post-MI remodelling.

Retroviral replicating vectors (RRVs) have achieved efficient tumor transduction and enhanced therapeutic benefit in a wide variety of cancer models. Here, we evaluated two different RRVs derived from amphotropic murine leukemia virus (AMLV) and gibbon ape leukemia virus (GALV), which utilize different cellular receptors (PiT-2 and PiT-1, respectively) for viral entry, in human osteosarcoma cells. Quantitative RT-PCR showed that low levels of expression of both receptors were observed in normal and non-malignant cells. However, high PiT-2 (for AMLV) and low PiT-1 (for GALV) expression was observed in most osteosarcoma cell lines. Accordingly, AMLV expressing the green fluorescent protein gene infected and replicated more efficiently than GALV in most osteosarcoma cell lines. Furthermore, RRVs expressing the cytosine deaminase prodrug activator gene showed differential cytotoxicity that correlated with the results of viral spread. AMLV-RRV-mediated prodrug activator gene therapy achieved significant inhibition of subcutaneous MG-63 tumor growth over GALV in nude mice. These data indicate that AMLV vectors predominate over GALV in human osteosarcoma cells. Moreover, our findings support the potential utility of the two RRVs in personalized cancer virotherapy on the basis of receptor expression.

Multiple research groups have shown that diet impacts the gut microbiome; however, variability in experimental design and quantitative assessment have made it challenging to assess the degree to which similar diets have reproducible effects across studies. Through an unbiased subject-level meta-analysis framework, we re-analyzed 27 dietary studies including 1,101 samples from rodents and humans. We demonstrate that a high-fat diet (HFD) reproducibly changes gut microbial community structure. Finer taxonomic analysis revealed that the most reproducible signals of a HFD are Lactococcus species, which we experimentally demonstrate to be common dietary contaminants. Additionally, a machine-learning approach defined a signature that predicts the dietary intake of mice and demonstrated that phylogenetic and gene-centric transformations of this model can be translated to humans. Together, these results demonstrate the utility of microbiome meta-analyses in identifying robust and reproducible features for mechanistic studies in preclinical models.

Heart failure continues to be a common and deadly sequela of myocardial infarction (MI). Despite strong evidence suggesting the importance of myocardial mechanics in cardiac remodelling, many MI studies still rely on two-dimensional analyses to estimate global left ventricular (LV) function. Here, we integrated four-dimensional ultrasound with three-dimensional strain mapping to longitudinally characterize LV mechanics within and around infarcts in order to study the post-MI remodelling process. To induce infarcts with varying severities, we separated 15 mice into three equal-sized groups: (i) sham, (ii) 30 min ischaemia-reperfusion, and (iii) permanent ligation of the left coronary artery. Four-dimensional ultrasound from a high-frequency small animal system was used to monitor changes in LV geometry, function and strain over 28 days. We reconstructed three-dimensional myocardial strain maps and showed that strain profiles at the infarct border followed a sigmoidal behaviour. We also identified that mice with mild remodelling had significantly higher strains in the infarcted myocardium than those with severe injury. Finally, we developed a new approach to non-invasively estimate infarct size from strain maps, which correlated well with histological results. Taken together, the presented work provides a thorough approach to quantify regional strain, an important component when assessing post-MI remodelling.

Liang Bua, the type locality of Homo floresiensis, is a limestone cave located in the western part of the Indonesian island of Flores. The relatively continuous stratigraphic sequence of the site spans the past ∼190 kyr and contains ∼275,000 taxonomically identifiable vertebrate skeletal elements, ∼80% of which belong to murine rodent taxa (i.e., rats). Six described genera are present at Liang Bua (Papagomys, Spelaeomys, Hooijeromys, Komodomys, Paulamys, and Rattus), one of which, Hooijeromys, is newly recorded in the site deposits, being previously known only from Early to Middle Pleistocene sites in central Flores. Measurements of the proximal femur (n = 10,212) and distal humerus (n = 1186) indicate five murine body size classes ranging from small (mouse-sized) to giant (common rabbit-sized) are present. The proportions of these five classes across successive stratigraphic units reveal two major changes in murine body size distribution due to significant shifts in the abundances of more open habitat-adapted medium-sized murines versus more closed habitat-adapted smaller-sized ones. One of these changes suggests a modest increase in available open habitats occurred ∼3 ka, likely the result of anthropogenic changes to the landscape related to farming by modern human populations. The other and more significant change occurred ∼60 ka suggesting a rapid shift from more open habitats to more closed conditions at this time. The abrupt reduction of medium-sized murines, along with the disappearance of H. floresiensis, Stegodon florensis insularis (an extinct proboscidean), Varanus komodoensis (Komodo dragon), Leptoptilos robustus (giant marabou stork), and Trigonoceps sp. (vulture) at Liang Bua ∼60-50 ka, is likely the consequence of these animals preferring and tracking more open habitats to elsewhere on the island. If correct, then the precise timing and nature of the extinction of H. floresiensis and its contemporaries must await new discoveries at Liang Bua or other

Liang Bua, the type locality of Homo floresiensis, is a limestone cave located in the western part of the Indonesian island of Flores. The relatively continuous stratigraphic sequence of the site spans the past ∼190 kyr and contains ∼275,000 taxonomically identifiable vertebrate skeletal elements, ∼80% of which belong to murine rodent taxa (i.e., rats). Six described genera are present at Liang Bua (Papagomys, Spelaeomys, Hooijeromys, Komodomys, Paulamys, and Rattus), one of which, Hooijeromys, is newly recorded in the site deposits, being previously known only from Early to Middle Pleistocene sites in central Flores. Measurements of the proximal femur (n = 10,212) and distal humerus (n = 1186) indicate five murine body size classes ranging from small (mouse-sized) to giant (common rabbit-sized) are present. The proportions of these five classes across successive stratigraphic units reveal two major changes in murine body size distribution due to significant shifts in the abundances of more open habitat-adapted medium-sized murines versus more closed habitat-adapted smaller-sized ones. One of these changes suggests a modest increase in available open habitats occurred ∼3 ka, likely the result of anthropogenic changes to the landscape related to farming by modern human populations. The other and more significant change occurred ∼60 ka suggesting a rapid shift from more open habitats to more closed conditions at this time. The abrupt reduction of medium-sized murines, along with the disappearance of H. floresiensis, Stegodon florensis insularis (an extinct proboscidean), Varanus komodoensis (Komodo dragon), Leptoptilos robustus (giant marabou stork), and Trigonoceps sp. (vulture) at Liang Bua ∼60-50 ka, is likely the consequence of these animals preferring and tracking more open habitats to elsewhere on the island. If correct, then the precise timing and nature of the extinction of H. floresiensis and its contemporaries must await new discoveries at Liang Bua or other

Giardia lamblia, a protozoan intestinal parasite is able to evade or suppress defense mechanisms, such as innate response. Antigen presenting cells (APC) like dendritic cells may orchestrate an immune response and support a more effective adaptive defense. Conjugated linoleic acid (CLA), a dietary lipid, has shown biological activity on APC. The aim was to evaluate the effect of CLA in intestinal innate response measuring the frequency of mucosal APC populations in Giardia lamblia murine infection. Method The giardiasis infection model was established in C3H/HeN mice (n=32), and parasite load was followed at 0, 2, 6 and 8 days post infection (dpi). APC obtained from small intestine by enzymatic digestion were assessed by flow cytometry. Oral supplementation with CLA (50:50 of cis-9, trans-11-CLA and trans-10, cis-12-CLA isomers) and placebo control begun three days before infection and continued during first week post infection. Results Infection kinetics showed a peak of trophozoites at 6 dpi in acute phase. Parasite load was lower in CLA group in comparison with control infected group (p(0.05). Conjugated linoleic acid stimulates innate immune response, percentage of intestinal CD11chiMHC-IIhi increases after 2 dpi, meanwhile CD11chiMHC-IIhiCD103+ APC was higher after three days of CLA supplementation than control (p(0.05). Also, the CD11c+ F4/80+ population shown a percentage increases after 6 and 8 dpi by the effect of treatment and time of infection (p<0.05). Conclusion In conclusion CLA increased percentages of small intestine APC phenotypes CD11chiMHCIIhi, CD11chiMHCIIhiCD103+ and CD11c+ F4/80+, and reduced G. lamblia parasitic load. Further studies are needed to elucidate potential mechanisms involved in CLA immunomodulatory effect and its contribution in adaptive immune response., Giardia lamblia es un parásito intestinal de prevalencia mundial. Es capaz de evadir los mecanismos de defensa del hospedero. Las células presentadoras de antígeno (CPA), como las células dendríticas y macrófagos, orquestan la respuesta inmune y favorecen el desarrollo de una respuesta adaptativa más robusta y eficiente, y tienen un papel clave en la respuesta inmune de mucosas en intestino. Algunos componentes de la dieta pueden modular la capacidad de respuesta del sistema inmune, como ejemplo el ácido linoleico conjugado, un lípido de origen dietario que presenta actividad biológica sobre células del linaje monocito fagocito. El objetivo del estudio fue evaluar la actividad del CLA sobre la respuesta inmune innata intestinal en un modelo murino de infección con Giardia lamblia. Método El modelo de infección se estableció en los ratones C3H/HeN (n=32). Se diseñó un bioensayo experimental con dos grupos de animales uno suplementado con CLA (mezcla de isómeros 50:50 de 9-cis, 11-trans y 10-trans, 12-cis) y otro control con PBS. La suplementación oral se dio por 11 días, 3 días previo a la infección, y continuó por 8 días más después de la infección. Se sacrificaron cuatro animales de cada grupo a los 0, 2, 6 y 8 días post infección (dpi) para conteo de trofozoítos y obtención de las células presentadoras de antígeno, a partir de digestión enzimática del intestino delgado. La fenotipificación de poblaciones se realizó por citometría de flujo. Resultados La cinética de infección mostró un pico en el número de trofozoítos a los 6 dpi en la fase aguda. La carga parasitaria fue menor en el grupo con CLA comparado con el grupo control (p(0.05). Los porcentajes de las poblaciones de APC intestinales CD11chiMHC-IIhi se incrementaron a los 2 dpi y CD11chiMHC-IIhiCD103+ de forma temprana después de 3 días de suplementación con CLA, comparado con el grupo control (p(0.05). La población de células CD11c+ F4/80+, también mostró un incremento por efecto del tratamiento y del tiem

Giardia lamblia, a protozoan intestinal parasite is able to evade or suppress defense mechanisms, such as innate response. Antigen presenting cells (APC) like dendritic cells may orchestrate an immune response and support a more effective adaptive defense. Conjugated linoleic acid (CLA), a dietary lipid, has shown biological activity on APC. The aim was to evaluate the effect of CLA in intestinal innate response measuring the frequency of mucosal APC populations in Giardia lamblia murine infection. Method The giardiasis infection model was established in C3H/HeN mice (n=32), and parasite load was followed at 0, 2, 6 and 8 days post infection (dpi). APC obtained from small intestine by enzymatic digestion were assessed by flow cytometry. Oral supplementation with CLA (50:50 of cis-9, trans-11-CLA and trans-10, cis-12-CLA isomers) and placebo control begun three days before infection and continued during first week post infection. Results Infection kinetics showed a peak of trophozoites at 6 dpi in acute phase. Parasite load was lower in CLA group in comparison with control infected group (p(0.05). Conjugated linoleic acid stimulates innate immune response, percentage of intestinal CD11chiMHC-IIhi increases after 2 dpi, meanwhile CD11chiMHC-IIhiCD103+ APC was higher after three days of CLA supplementation than control (p(0.05). Also, the CD11c+ F4/80+ population shown a percentage increases after 6 and 8 dpi by the effect of treatment and time of infection (p<0.05). Conclusion In conclusion CLA increased percentages of small intestine APC phenotypes CD11chiMHCIIhi, CD11chiMHCIIhiCD103+ and CD11c+ F4/80+, and reduced G. lamblia parasitic load. Further studies are needed to elucidate potential mechanisms involved in CLA immunomodulatory effect and its contribution in adaptive immune response., Giardia lamblia es un parásito intestinal de prevalencia mundial. Es capaz de evadir los mecanismos de defensa del hospedero. Las células presentadoras de antígeno (CPA), como las células dendríticas y macrófagos, orquestan la respuesta inmune y favorecen el desarrollo de una respuesta adaptativa más robusta y eficiente, y tienen un papel clave en la respuesta inmune de mucosas en intestino. Algunos componentes de la dieta pueden modular la capacidad de respuesta del sistema inmune, como ejemplo el ácido linoleico conjugado, un lípido de origen dietario que presenta actividad biológica sobre células del linaje monocito fagocito. El objetivo del estudio fue evaluar la actividad del CLA sobre la respuesta inmune innata intestinal en un modelo murino de infección con Giardia lamblia. Método El modelo de infección se estableció en los ratones C3H/HeN (n=32). Se diseñó un bioensayo experimental con dos grupos de animales uno suplementado con CLA (mezcla de isómeros 50:50 de 9-cis, 11-trans y 10-trans, 12-cis) y otro control con PBS. La suplementación oral se dio por 11 días, 3 días previo a la infección, y continuó por 8 días más después de la infección. Se sacrificaron cuatro animales de cada grupo a los 0, 2, 6 y 8 días post infección (dpi) para conteo de trofozoítos y obtención de las células presentadoras de antígeno, a partir de digestión enzimática del intestino delgado. La fenotipificación de poblaciones se realizó por citometría de flujo. Resultados La cinética de infección mostró un pico en el número de trofozoítos a los 6 dpi en la fase aguda. La carga parasitaria fue menor en el grupo con CLA comparado con el grupo control (p(0.05). Los porcentajes de las poblaciones de APC intestinales CD11chiMHC-IIhi se incrementaron a los 2 dpi y CD11chiMHC-IIhiCD103+ de forma temprana después de 3 días de suplementación con CLA, comparado con el grupo control (p(0.05). La población de células CD11c+ F4/80+, también mostró un incremento por efecto del tratamiento y del tiem

Giardia lamblia, a protozoan intestinal parasite is able to evade or suppress defense mechanisms, such as innate response. Antigen presenting cells (APC) like dendritic cells may orchestrate an immune response and support a more effective adaptive defense. Conjugated linoleic acid (CLA), a dietary lipid, has shown biological activity on APC. The aim was to evaluate the effect of CLA in intestinal innate response measuring the frequency of mucosal APC populations in Giardia lamblia murine infection. Method The giardiasis infection model was established in C3H/HeN mice (n=32), and parasite load was followed at 0, 2, 6 and 8 days post infection (dpi). APC obtained from small intestine by enzymatic digestion were assessed by flow cytometry. Oral supplementation with CLA (50:50 of cis-9, trans-11-CLA and trans-10, cis-12-CLA isomers) and placebo control begun three days before infection and continued during first week post infection. Results Infection kinetics showed a peak of trophozoites at 6 dpi in acute phase. Parasite load was lower in CLA group in comparison with control infected group (p(0.05). Conjugated linoleic acid stimulates innate immune response, percentage of intestinal CD11chiMHC-IIhi increases after 2 dpi, meanwhile CD11chiMHC-IIhiCD103+ APC was higher after three days of CLA supplementation than control (p(0.05). Also, the CD11c+ F4/80+ population shown a percentage increases after 6 and 8 dpi by the effect of treatment and time of infection (p<0.05). Conclusion In conclusion CLA increased percentages of small intestine APC phenotypes CD11chiMHCIIhi, CD11chiMHCIIhiCD103+ and CD11c+ F4/80+, and reduced G. lamblia parasitic load. Further studies are needed to elucidate potential mechanisms involved in CLA immunomodulatory effect and its contribution in adaptive immune response., Giardia lamblia es un parásito intestinal de prevalencia mundial. Es capaz de evadir los mecanismos de defensa del hospedero. Las células presentadoras de antígeno (CPA), como las células dendríticas y macrófagos, orquestan la respuesta inmune y favorecen el desarrollo de una respuesta adaptativa más robusta y eficiente, y tienen un papel clave en la respuesta inmune de mucosas en intestino. Algunos componentes de la dieta pueden modular la capacidad de respuesta del sistema inmune, como ejemplo el ácido linoleico conjugado, un lípido de origen dietario que presenta actividad biológica sobre células del linaje monocito fagocito. El objetivo del estudio fue evaluar la actividad del CLA sobre la respuesta inmune innata intestinal en un modelo murino de infección con Giardia lamblia. Método El modelo de infección se estableció en los ratones C3H/HeN (n=32). Se diseñó un bioensayo experimental con dos grupos de animales uno suplementado con CLA (mezcla de isómeros 50:50 de 9-cis, 11-trans y 10-trans, 12-cis) y otro control con PBS. La suplementación oral se dio por 11 días, 3 días previo a la infección, y continuó por 8 días más después de la infección. Se sacrificaron cuatro animales de cada grupo a los 0, 2, 6 y 8 días post infección (dpi) para conteo de trofozoítos y obtención de las células presentadoras de antígeno, a partir de digestión enzimática del intestino delgado. La fenotipificación de poblaciones se realizó por citometría de flujo. Resultados La cinética de infección mostró un pico en el número de trofozoítos a los 6 dpi en la fase aguda. La carga parasitaria fue menor en el grupo con CLA comparado con el grupo control (p(0.05). Los porcentajes de las poblaciones de APC intestinales CD11chiMHC-IIhi se incrementaron a los 2 dpi y CD11chiMHC-IIhiCD103+ de forma temprana después de 3 días de suplementación con CLA, comparado con el grupo control (p(0.05). La población de células CD11c+ F4/80+, también mostró un incremento por efecto del tratamiento y del tiem

Giardia lamblia, a protozoan intestinal parasite is able to evade or suppress defense mechanisms, such as innate response. Antigen presenting cells (APC) like dendritic cells may orchestrate an immune response and support a more effective adaptive defense. Conjugated linoleic acid (CLA), a dietary lipid, has shown biological activity on APC. The aim was to evaluate the effect of CLA in intestinal innate response measuring the frequency of mucosal APC populations in Giardia lamblia murine infection. Method The giardiasis infection model was established in C3H/HeN mice (n=32), and parasite load was followed at 0, 2, 6 and 8 days post infection (dpi). APC obtained from small intestine by enzymatic digestion were assessed by flow cytometry. Oral supplementation with CLA (50:50 of cis-9, trans-11-CLA and trans-10, cis-12-CLA isomers) and placebo control begun three days before infection and continued during first week post infection. Results Infection kinetics showed a peak of trophozoites at 6 dpi in acute phase. Parasite load was lower in CLA group in comparison with control infected group (p(0.05). Conjugated linoleic acid stimulates innate immune response, percentage of intestinal CD11chiMHC-IIhi increases after 2 dpi, meanwhile CD11chiMHC-IIhiCD103+ APC was higher after three days of CLA supplementation than control (p(0.05). Also, the CD11c+ F4/80+ population shown a percentage increases after 6 and 8 dpi by the effect of treatment and time of infection (p<0.05). Conclusion In conclusion CLA increased percentages of small intestine APC phenotypes CD11chiMHCIIhi, CD11chiMHCIIhiCD103+ and CD11c+ F4/80+, and reduced G. lamblia parasitic load. Further studies are needed to elucidate potential mechanisms involved in CLA immunomodulatory effect and its contribution in adaptive immune response., Giardia lamblia es un parásito intestinal de prevalencia mundial. Es capaz de evadir los mecanismos de defensa del hospedero. Las células presentadoras de antígeno (CPA), como las células dendríticas y macrófagos, orquestan la respuesta inmune y favorecen el desarrollo de una respuesta adaptativa más robusta y eficiente, y tienen un papel clave en la respuesta inmune de mucosas en intestino. Algunos componentes de la dieta pueden modular la capacidad de respuesta del sistema inmune, como ejemplo el ácido linoleico conjugado, un lípido de origen dietario que presenta actividad biológica sobre células del linaje monocito fagocito. El objetivo del estudio fue evaluar la actividad del CLA sobre la respuesta inmune innata intestinal en un modelo murino de infección con Giardia lamblia. Método El modelo de infección se estableció en los ratones C3H/HeN (n=32). Se diseñó un bioensayo experimental con dos grupos de animales uno suplementado con CLA (mezcla de isómeros 50:50 de 9-cis, 11-trans y 10-trans, 12-cis) y otro control con PBS. La suplementación oral se dio por 11 días, 3 días previo a la infección, y continuó por 8 días más después de la infección. Se sacrificaron cuatro animales de cada grupo a los 0, 2, 6 y 8 días post infección (dpi) para conteo de trofozoítos y obtención de las células presentadoras de antígeno, a partir de digestión enzimática del intestino delgado. La fenotipificación de poblaciones se realizó por citometría de flujo. Resultados La cinética de infección mostró un pico en el número de trofozoítos a los 6 dpi en la fase aguda. La carga parasitaria fue menor en el grupo con CLA comparado con el grupo control (p(0.05). Los porcentajes de las poblaciones de APC intestinales CD11chiMHC-IIhi se incrementaron a los 2 dpi y CD11chiMHC-IIhiCD103+ de forma temprana después de 3 días de suplementación con CLA, comparado con el grupo control (p(0.05). La población de células CD11c+ F4/80+, también mostró un incremento por efecto del tratamiento y del tiem

Giardia lamblia, a protozoan intestinal parasite is able to evade or suppress defense mechanisms, such as innate response. Antigen presenting cells (APC) like dendritic cells may orchestrate an immune response and support a more effective adaptive defense. Conjugated linoleic acid (CLA), a dietary lipid, has shown biological activity on APC. The aim was to evaluate the effect of CLA in intestinal innate response measuring the frequency of mucosal APC populations in Giardia lamblia murine infection. Method The giardiasis infection model was established in C3H/HeN mice (n=32), and parasite load was followed at 0, 2, 6 and 8 days post infection (dpi). APC obtained from small intestine by enzymatic digestion were assessed by flow cytometry. Oral supplementation with CLA (50:50 of cis-9, trans-11-CLA and trans-10, cis-12-CLA isomers) and placebo control begun three days before infection and continued during first week post infection. Results Infection kinetics showed a peak of trophozoites at 6 dpi in acute phase. Parasite load was lower in CLA group in comparison with control infected group (p(0.05). Conjugated linoleic acid stimulates innate immune response, percentage of intestinal CD11chiMHC-IIhi increases after 2 dpi, meanwhile CD11chiMHC-IIhiCD103+ APC was higher after three days of CLA supplementation than control (p(0.05). Also, the CD11c+ F4/80+ population shown a percentage increases after 6 and 8 dpi by the effect of treatment and time of infection (p<0.05). Conclusion In conclusion CLA increased percentages of small intestine APC phenotypes CD11chiMHCIIhi, CD11chiMHCIIhiCD103+ and CD11c+ F4/80+, and reduced G. lamblia parasitic load. Further studies are needed to elucidate potential mechanisms involved in CLA immunomodulatory effect and its contribution in adaptive immune response., Giardia lamblia es un parásito intestinal de prevalencia mundial. Es capaz de evadir los mecanismos de defensa del hospedero. Las células presentadoras de antígeno (CPA), como las células dendríticas y macrófagos, orquestan la respuesta inmune y favorecen el desarrollo de una respuesta adaptativa más robusta y eficiente, y tienen un papel clave en la respuesta inmune de mucosas en intestino. Algunos componentes de la dieta pueden modular la capacidad de respuesta del sistema inmune, como ejemplo el ácido linoleico conjugado, un lípido de origen dietario que presenta actividad biológica sobre células del linaje monocito fagocito. El objetivo del estudio fue evaluar la actividad del CLA sobre la respuesta inmune innata intestinal en un modelo murino de infección con Giardia lamblia. Método El modelo de infección se estableció en los ratones C3H/HeN (n=32). Se diseñó un bioensayo experimental con dos grupos de animales uno suplementado con CLA (mezcla de isómeros 50:50 de 9-cis, 11-trans y 10-trans, 12-cis) y otro control con PBS. La suplementación oral se dio por 11 días, 3 días previo a la infección, y continuó por 8 días más después de la infección. Se sacrificaron cuatro animales de cada grupo a los 0, 2, 6 y 8 días post infección (dpi) para conteo de trofozoítos y obtención de las células presentadoras de antígeno, a partir de digestión enzimática del intestino delgado. La fenotipificación de poblaciones se realizó por citometría de flujo. Resultados La cinética de infección mostró un pico en el número de trofozoítos a los 6 dpi en la fase aguda. La carga parasitaria fue menor en el grupo con CLA comparado con el grupo control (p(0.05). Los porcentajes de las poblaciones de APC intestinales CD11chiMHC-IIhi se incrementaron a los 2 dpi y CD11chiMHC-IIhiCD103+ de forma temprana después de 3 días de suplementación con CLA, comparado con el grupo control (p(0.05). La población de células CD11c+ F4/80+, también mostró un incremento por efecto del tratamiento y del tiem

Giardia lamblia, a protozoan intestinal parasite is able to evade or suppress defense mechanisms, such as innate response. Antigen presenting cells (APC) like dendritic cells may orchestrate an immune response and support a more effective adaptive defense. Conjugated linoleic acid (CLA), a dietary lipid, has shown biological activity on APC. The aim was to evaluate the effect of CLA in intestinal innate response measuring the frequency of mucosal APC populations in Giardia lamblia murine infection. Method The giardiasis infection model was established in C3H/HeN mice (n=32), and parasite load was followed at 0, 2, 6 and 8 days post infection (dpi). APC obtained from small intestine by enzymatic digestion were assessed by flow cytometry. Oral supplementation with CLA (50:50 of cis-9, trans-11-CLA and trans-10, cis-12-CLA isomers) and placebo control begun three days before infection and continued during first week post infection. Results Infection kinetics showed a peak of trophozoites at 6 dpi in acute phase. Parasite load was lower in CLA group in comparison with control infected group (p(0.05). Conjugated linoleic acid stimulates innate immune response, percentage of intestinal CD11chiMHC-IIhi increases after 2 dpi, meanwhile CD11chiMHC-IIhiCD103+ APC was higher after three days of CLA supplementation than control (p(0.05). Also, the CD11c+ F4/80+ population shown a percentage increases after 6 and 8 dpi by the effect of treatment and time of infection (p<0.05). Conclusion In conclusion CLA increased percentages of small intestine APC phenotypes CD11chiMHCIIhi, CD11chiMHCIIhiCD103+ and CD11c+ F4/80+, and reduced G. lamblia parasitic load. Further studies are needed to elucidate potential mechanisms involved in CLA immunomodulatory effect and its contribution in adaptive immune response., Giardia lamblia es un parásito intestinal de prevalencia mundial. Es capaz de evadir los mecanismos de defensa del hospedero. Las células presentadoras de antígeno (CPA), como las células dendríticas y macrófagos, orquestan la respuesta inmune y favorecen el desarrollo de una respuesta adaptativa más robusta y eficiente, y tienen un papel clave en la respuesta inmune de mucosas en intestino. Algunos componentes de la dieta pueden modular la capacidad de respuesta del sistema inmune, como ejemplo el ácido linoleico conjugado, un lípido de origen dietario que presenta actividad biológica sobre células del linaje monocito fagocito. El objetivo del estudio fue evaluar la actividad del CLA sobre la respuesta inmune innata intestinal en un modelo murino de infección con Giardia lamblia. Método El modelo de infección se estableció en los ratones C3H/HeN (n=32). Se diseñó un bioensayo experimental con dos grupos de animales uno suplementado con CLA (mezcla de isómeros 50:50 de 9-cis, 11-trans y 10-trans, 12-cis) y otro control con PBS. La suplementación oral se dio por 11 días, 3 días previo a la infección, y continuó por 8 días más después de la infección. Se sacrificaron cuatro animales de cada grupo a los 0, 2, 6 y 8 días post infección (dpi) para conteo de trofozoítos y obtención de las células presentadoras de antígeno, a partir de digestión enzimática del intestino delgado. La fenotipificación de poblaciones se realizó por citometría de flujo. Resultados La cinética de infección mostró un pico en el número de trofozoítos a los 6 dpi en la fase aguda. La carga parasitaria fue menor en el grupo con CLA comparado con el grupo control (p(0.05). Los porcentajes de las poblaciones de APC intestinales CD11chiMHC-IIhi se incrementaron a los 2 dpi y CD11chiMHC-IIhiCD103+ de forma temprana después de 3 días de suplementación con CLA, comparado con el grupo control (p(0.05). La población de células CD11c+ F4/80+, también mostró un incremento por efecto del tratamiento y del tiem

We compared prophylactic or continuous therapy with the investigational drug VT-1161 to that with posaconazole in treating murine mucormycosis due to Rhizopus arrhizus var. arrhizus In the prophylaxis studies, only VT-1161 resulted in improved survival and lowered tissue fungal burden of immunosuppressed infected mice. In the continuous therapy, VT-1161 outperformed posaconazole in prolonging mouse survival time despite its comparable effect in lowering tissue fungal burden. These results support the further development of VT-1161 against mucormycosis.

BACKGROUND: Pregnancy induces adaptations in maternal metabolism to meet the increased need for nutrients by the placenta and fetus. Creatine is an important intracellular metabolite obtained from the diet and also synthesised endogenously. Experimental evidence suggests that the fetus relies on a maternal supply of creatine for much of gestation. However, the impact of pregnancy on maternal creatine homeostasis is unclear. We hypothesise that alteration of maternal creatine homeostasis occurs during pregnancy to ensure adequate levels of this essential substrate are available for maternal tissues, the placenta and fetus. This study aimed to describe maternal creatine homeostasis from mid to late gestation in the precocial spiny mouse. METHODS: Plasma creatine concentration and urinary excretion were measured from mid to late gestation in pregnant (n = 8) and age-matched virgin female spiny mice (n = 6). At term, body composition and organ weights were assessed and tissue total creatine content determined. mRNA expression of the creatine synthesising enzymes arginine:glycine amidinotransferase (AGAT) and guanidinoacetate methyltransferase (GAMT), and the creatine transporter (CrT1) were assessed by RT-qPCR. Protein expression of AGAT and GAMT was also assessed by western blot analysis. RESULTS: Plasma creatine and renal creatine excretion decreased significantly from mid to late gestation (P < 0.001, P < 0.05, respectively). Pregnancy resulted in increased lean tissue (P < 0.01), kidney (P < 0.01), liver (P < 0.01) and heart (P < 0.05) mass at term. CrT1 expression was increased in the heart (P < 0.05) and skeletal muscle (P < 0.05) at term compared to non-pregnant tissues, and creatine content of the heart (P < 0.05) and kidney (P < 0.001) were also increased at this time. CrT1 mRNA expression was down-regulated in the liver (<0.01) and brain (<0.01) of pregnant spiny mice at term. Renal AGAT mRNA (P < 0.01) and protein (P < 0.05) expression were both significantly

Pigmentation plays a protective role against damage caused by ultraviolet (UV) irradiation. Humans with fair skin and light hair have a higher susceptibility to UV-induced DNA damage that can lead to the development of skin cancers. The melanocytes found in the skin and hair follicles depend on different signaling molecules for their proper development and pigment production. α-Melanocyte Stimulating Hormone (α-msh) binds to the Melanocortin 1 receptor (Mc1r) to regulate pigment production and the switch between eumelanin and pheomelanin. Lethal yellow mice (Ay) overexpress the agouti signaling protein, which inhibits the binding of α-msh, resulting in a yellow coat color phenotype. Endothelin 3 (Edn3) encodes for a ligand involved in melanocyte development by regulating the differentiation, proliferation and migration of melanocyte precursors. A tetracycline inducible transgenic mouse in which Edn3 was placed under the keratin 5 promoter (K5-tTA;TRE-Edn3-lacZ) displays a hyperpigmentation phenotype due to the accumulation of melanocytes in the skin and an increase in hair pigment. Comparative analysis of dorsal hairs from Ay and Ay; K5-tTA;TRE-Edn3-lacZ mice using high performance liquid chromatography showed that transgenic Edn3 expression significantly increased both eumelanin and pheomelanin. No significant difference in the number of follicular melanocytes between Edn3 transgenic and non-transgenic mice was evidenced by immunofluorescence using an antibody against Tyrosinase related protein 1. Gene expression analysis of hair follicles showed that Edn3 upregulates the expression of melanogenic genes. Deactivation of transgenic Edn3 is possible with doxycycline (dox) treatment. To test if transgenic Edn3 expression is required to rescue and maintain a dark pigmentation phenotype in Ay mice, dox was administered during embryonic and postnatal development to manipulate transgenic Edn3 expression. Results showed that transgenic Edn3 expression is required to mainta

BACKGROUND: Pregnancy induces adaptations in maternal metabolism to meet the increased need for nutrients by the placenta and fetus. Creatine is an important intracellular metabolite obtained from the diet and also synthesised endogenously. Experimental evidence suggests that the fetus relies on a maternal supply of creatine for much of gestation. However, the impact of pregnancy on maternal creatine homeostasis is unclear. We hypothesise that alteration of maternal creatine homeostasis occurs during pregnancy to ensure adequate levels of this essential substrate are available for maternal tissues, the placenta and fetus. This study aimed to describe maternal creatine homeostasis from mid to late gestation in the precocial spiny mouse. METHODS: Plasma creatine concentration and urinary excretion were measured from mid to late gestation in pregnant (n = 8) and age-matched virgin female spiny mice (n = 6). At term, body composition and organ weights were assessed and tissue total creatine content determined. mRNA expression of the creatine synthesising enzymes arginine:glycine amidinotransferase (AGAT) and guanidinoacetate methyltransferase (GAMT), and the creatine transporter (CrT1) were assessed by RT-qPCR. Protein expression of AGAT and GAMT was also assessed by western blot analysis. RESULTS: Plasma creatine and renal creatine excretion decreased significantly from mid to late gestation (P < 0.001, P < 0.05, respectively). Pregnancy resulted in increased lean tissue (P < 0.01), kidney (P < 0.01), liver (P < 0.01) and heart (P < 0.05) mass at term. CrT1 expression was increased in the heart (P < 0.05) and skeletal muscle (P < 0.05) at term compared to non-pregnant tissues, and creatine content of the heart (P < 0.05) and kidney (P < 0.001) were also increased at this time. CrT1 mRNA expression was down-regulated in the liver (<0.01) and brain (<0.01) of pregnant spiny mice at term. Renal AGAT mRNA (P < 0.01) and protein (P < 0.05) expression were both significantly

We compared prophylactic or continuous therapy with the investigational drug VT-1161 to that with posaconazole in treating murine mucormycosis due to Rhizopus arrhizus var. arrhizus In the prophylaxis studies, only VT-1161 resulted in improved survival and lowered tissue fungal burden of immunosuppressed infected mice. In the continuous therapy, VT-1161 outperformed posaconazole in prolonging mouse survival time despite its comparable effect in lowering tissue fungal burden. These results support the further development of VT-1161 against mucormycosis.

Active participation of endogenous retroviruses (ERVs) in disease processes has been exemplified by the finding that the HERV (human ERV)-W envelope protein is involved in the pathogenesis of multiple sclerosis, an autoimmune disease. We also demonstrated that injury-elicited stressors alter the expression of murine ERVs (MuERVs), both murine leukemia virus-type and mouse mammary tumor virus (MMTV)-type (MMTV-MuERV). In this study, to evaluate MMTV-MuERVs' responses to stress (e.g., injury, infection)-elicited systemic glucocorticoid (GC) levels, we examined the GC-stress response of 64 MMTV-MuERV promoters isolated from the genomes of 23 mouse strains. All 64 promoters responded to treatment with a synthetic GC, dexamethasone (DEX), at a wide range from a 0.6- to 85.7-fold increase in reporter activity compared to no treatment. An analysis of the 10 lowest and 10 highest DEX responders revealed specific promoter elements exclusively present in either the three lowest or the two highest responders. Each promoter had a unique profile of transcription regulatory elements and the glucocorticoid response element (GRE) was identified in all promoters with the number of GREs ranging from 2 to 7. The three lowest DEX responders were the only promoters with two GREs. The findings from this study suggest that certain MMTV-MuERVs are more responsive to stress-elicited systemic GC elevation compared to the others. The mouse strain-specific genomic MMTV-MuERV profiles and individual MMTV-MuERVs' differential responses to GC-stress might explain, at least in part, the variable inflammatory responses to injury and/or infection, often observed among different mouse strains. This article is part of a Special Issue entitled: Immune and Metabolic Alterations in Trauma and Sepsis edited by Dr. Raghavan Raju.

In recent years, many spore-forming commensal Clostridia found in the gut have been discovered to promote host physiology, immune development, and protection against infections. We provide a detailed protocol for rapid enrichment of spore-forming bacteria from murine intestine. Briefly, contents from the intestinal cecum are collected aerobically, diluted and finally treated with chloroform to enrich for Clostridia spores.

We assessed prophylactic or continuous therapy of isavuconazole, posaconazole, or voriconazole in treating pulmonary murine mucormycosis. In the prophylaxis studies, only isavuconazole treatment resulted in significantly improved survival and lowered tissue fungal burden of immunosuppressed mice infected with Rhizopus delemar. In the continuous treatment studies, isavuconazole and posaconazole, but not voriconazole, equally prolonged survival time and lowered tissue fungal burden compared to placebo-treated mice. These results support the use of isavuconazole and posaconazole in prophylaxis treatment.

Background/Purpose: Glucocorticoids (GC) have been used to treat inflammatory disease for more than six decades, but the broad spectrum of therapeutic effects is accompanied by significant metabolic adverse effects. Recent studies suggest that glucocorticoid-induced leucine zipper (GILZ) mimics the anti-inflammatory effects of GC, including inhibiting the activation of macrophage, B cells, Th1 and Th17 cells, but lacks GC adverse effects. However, little is known about molecular mechanisms of GILZ action that could empower drug discovery. Method(s): Expression of GILZ and acetylated GILZ was measured by Western blotting in murine bone marrow-derived macrophages. WT and mutated GILZ plasmids were co-transfected with an NF-kappaB luciferase reporter into human microvascular endothelial cells (HMECs). IL-6 and MCP-1 were measured by ELISA. NF-kappaB activity was measured using a NF-kappaB luciferase reporter assay. Interactions between GILZ and p65/NF-kappaB were detected by Fluorescence lifetime imaging (FLIM)/ Fluorescence Resonance Energy Transfer (FRET). Result(s): GILZ acetylation was detected in macrophages, and TSA, a deacetylase inhibitor, increased GILZ acetylation in a time-dependent manner. WT GILZ was constructed in the pcDNA3.1 expression plasmid and overexpression of GILZ significantly inhibited NF-kappaB activity and IL-6 and MCP-1 expression. Using an acetylation substrate-binding site alignment approach, we predicted three potential GILZ acetylation sites at K37, K77 and K108. Acetylation of GILZ was lost when K77, but not K37 or K108, was mutated to arginine (K77R). Mutation at K77 aborgated GILZ inhibition of NF-kappaB activity, and IL-6 and MCP-1 expression. Co-IP and FLIM/FRET demonstrated that GILZ mutation at K77 prevented interaction with p65/NF-kappaB. Conclusion(s): Acetylation of GILZ at K77 is required for its interaction with NF-kappaB and anti-inflammatory effects. This suggests a potential target for enhancing the anti-inflammatory effect

Background/Purpose: Glucocorticoids (GC) have been used to treat inflammatory disease for more than six decades, but the broad spectrum of therapeutic effects is accompanied by significant metabolic adverse effects. Recent studies suggest that glucocorticoid-induced leucine zipper (GILZ) mimics the anti-inflammatory effects of GC, including inhibiting the activation of macrophage, B cells, Th1 and Th17 cells, but lacks GC adverse effects. However, little is known about molecular mechanisms of GILZ action that could empower drug discovery. Method(s): Expression of GILZ and acetylated GILZ was measured by Western blotting in murine bone marrow-derived macrophages. WT and mutated GILZ plasmids were co-transfected with an NF-kappaB luciferase reporter into human microvascular endothelial cells (HMECs). IL-6 and MCP-1 were measured by ELISA. NF-kappaB activity was measured using a NF-kappaB luciferase reporter assay. Interactions between GILZ and p65/NF-kappaB were detected by Fluorescence lifetime imaging (FLIM)/ Fluorescence Resonance Energy Transfer (FRET). Result(s): GILZ acetylation was detected in macrophages, and TSA, a deacetylase inhibitor, increased GILZ acetylation in a time-dependent manner. WT GILZ was constructed in the pcDNA3.1 expression plasmid and overexpression of GILZ significantly inhibited NF-kappaB activity and IL-6 and MCP-1 expression. Using an acetylation substrate-binding site alignment approach, we predicted three potential GILZ acetylation sites at K37, K77 and K108. Acetylation of GILZ was lost when K77, but not K37 or K108, was mutated to arginine (K77R). Mutation at K77 aborgated GILZ inhibition of NF-kappaB activity, and IL-6 and MCP-1 expression. Co-IP and FLIM/FRET demonstrated that GILZ mutation at K77 prevented interaction with p65/NF-kappaB. Conclusion(s): Acetylation of GILZ at K77 is required for its interaction with NF-kappaB and anti-inflammatory effects. This suggests a potential target for enhancing the anti-inflammatory effect

Several studies have been conducted to check the prevalence of methicillin-resistant strains of Staphylococcus aureus (MRSA) in animals and animal-derived food products but limited data are available regarding their virulence and associated gene expression profile. In the present study, antibiotic resistance and virulence of MRSA and methicillin-sensitive S. aureus animal isolates were determined in vitro by agar dilution, biofilm formation, adhesion, invasion and intracellular survivability assays. In addition, the pathogenicity of these isolates was examined in a murine model of S. aureus sepsis. MRSA1679a, a strain isolated from chicken, was observed to be highly virulent, in cell culture and in mouse model, and exhibited extensive resistant profile. Comparative gene expression profile of MRSA1679a and the reference human MRSA strain (ATCC 29213) was performed using Illumina-based transcriptome and RT-qPCR analyses. Several virulence elements including 22 toxin genes were detected in MRSA animal-isolate. In addition, we observed enhanced expression of crucial virulence regulators, such as sarA and KdpDE in MRSA animal-isolate compared to the human isolate. Collectively, gene expression profile including several virulence and drug-resistance factors confirmed the unique and highly virulent determinants of the MRSA strain of poultry origin which warrants further attention due to significant threat to public health.

Laboratory strains of mice, both conventional and genetically engineered, have been introduced as critical components of a broad range of studies investigating normal and disease biology. Currently, the genetic identity of laboratory mice is primarily confirmed by surveying polymorphisms in selected sets of "conventional" genes and/or microsatellites in the absence of a single completely sequenced mouse genome. First, we examined variations in the genomic landscapes of transposable repetitive elements, named the TREome, in conventional and genetically engineered mouse strains using murine leukemia virus-type endogenous retroviruses (MLV-ERVs) as a probe. A survey of the genomes from 56 conventional strains revealed strain-specific TREome landscapes, and certain families (e.g., C57BL) of strains were discernible with defined patterns. Interestingly, the TREome landscapes of C3H/HeJ (toll-like receptor-4 [TLR4] mutant) inbred mice were different from its control C3H/HeOuJ (TLR4 wild-type) strain. In addition, a CD14 knock-out strain had a distinct TREome landscape compared to its control/backcross C57BL/6J strain. Second, an examination of superantigen (SAg, a "TREome gene") coding sequences of mouse mammary tumor virus-type ERVs in the genomes of the 46 conventional strains revealed a high diversity, suggesting a potential role of SAgs in strain-specific immune phenotypes. The findings from this study indicate that unexplored and intricate genomic variations exist in laboratory mouse strains, both conventional and genetically engineered. The TREome-based high-resolution genetics surveillance system for laboratory mice would contribute to efficient study design with quality control and accurate data interpretation. This genetics system can be easily adapted to other species ranging from plants to humans.

Several studies have been conducted to check the prevalence of methicillin-resistant strains of Staphylococcus aureus (MRSA) in animals and animal-derived food products but limited data are available regarding their virulence and associated gene expression profile. In the present study, antibiotic resistance and virulence of MRSA and methicillin-sensitive S. aureus animal isolates were determined in vitro by agar dilution, biofilm formation, adhesion, invasion and intracellular survivability assays. In addition, the pathogenicity of these isolates was examined in a murine model of S. aureus sepsis. MRSA1679a, a strain isolated from chicken, was observed to be highly virulent, in cell culture and in mouse model, and exhibited extensive resistant profile. Comparative gene expression profile of MRSA1679a and the reference human MRSA strain (ATCC 29213) was performed using Illumina-based transcriptome and RT-qPCR analyses. Several virulence elements including 22 toxin genes were detected in MRSA animal-isolate. In addition, we observed enhanced expression of crucial virulence regulators, such as sarA and KdpDE in MRSA animal-isolate compared to the human isolate. Collectively, gene expression profile including several virulence and drug-resistance factors confirmed the unique and highly virulent determinants of the MRSA strain of poultry origin which warrants further attention due to significant threat to public health.