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Indole-3-acetic acid (IAA), a crucial plant hormone, regulates diverse physiological processes. This study aimed to isolate and characterize IAA-producing bacteria from the chickpea (Cicer arietinum L.) rhizosphere and evaluate their effects on plant growth. From 54 rhizosphere samples, 118 bacteria (designated as GAC) were isolated and screened for IAA production using a Salkowski colorimetric assay, and Bergey's manual was used for biochemical identification. Isolates were grown under various conditions and in vitro screened for their growth promotion traits. A PCR investigation was performed for IAA and nitrogen-fixing genes, and evaluated for greenhouse conditions. Among them, 27 isolates produced IAA, with eight high producers selected. Morphological and biochemical identification classified the six isolates as Pseudomonas and the other two as Bacillus. Optimal conditions for IAA production were observed at 500 μg/ml tryptophan, 35 °C, and pH 7.0. A 48-h incubation was ideal for IAA production, except for GAC-34 and GAC-73, which required 72 h. All the isolates achieved optimal IAA levels with tryptone and sucrose as nitrogen and carbon sources, respectively. Moreover, all isolates showed nitrogen fixation ability, and the six isolates exhibited phosphate solubilization. PCR confirmed the amplification of nifH (300 bp), nifK (360 bp), and ipdC (1170 bp) genes. Greenhouse experiments demonstrated that eight selected isolates significantly enhanced chickpea growth parameters (p

Monostromatic algae, Monostroma and Gayralia, are taxonomically complex, with several nomenclatural changes over the past years owing to indistinguishable morphological features. Several Monostroma species were recently transferred to the genus Gayralia based on their ontogeny and molecular analysis. There is no record of the occurrence of Gayralia in Korea, whereas three species of Monostroma have been reported. We conducted a molecular analysis based on the nuclear ribosomal internal transcribed spacer region of the specimens from Jeju, Korea, that we considered to be included in the genus Monostroma. The specimens from this study and five sequences of Monostroma nitidum downloaded from the GenBank were conspecific with Gayralia kuroshiensis from Japan but distinct from the M. nitidum syntype from Australia. G. kuroshiensis has a single, expanded, laminar, monostromatic thallus that is light to mid-green. The thallus thickness in the marginal regions was similar to that of G. kuroshiensis from Japan, with an undulate shape and no marginal teeth. The cells in surface view were in groups of two, rectangular to polygonal in shape, with one to three pyrenoids per cell. Based on the morphological and molecular analyses, we report a new record of G. kuroshiensis in Korea.

Acral and mucosal melanomas are often driven by sequence variants in the KIT receptor tyrosine kinase, with nearly 40% harboring alterations in the KIT locus. Despite advances in the knowledge of KIT-mutated melanomas, little is known about the molecular reprogramming that occurs during KIT-mediated melanoma progression owing to the rarity of acral and mucosal melanomas and the lack of comprehensive biological tools and models. To this end, we used a murine model that allows us to ascertain the molecular underpinnings of the stages of cancer progression—transformation, tumorigenesis, immune engagement, and tumor escalation. We found dramatic increases in biosynthetic demands associated with the transformation stage, including DNA and RNA metabolism, leading to replication stress. Tumorigenesis was closely linked to neuronal and axonal development, likely necessary for invasion into the host. Immune engagement highlighted early immune excitation and rejection pathways, possibly triggered by abrupt neoantigen exposure. Finally, tumor escalation pathways proved consistent with immune evasion, with immune-related pathways becoming significantly downregulated. To our knowledge, it is previously unreported that these critical milestones needed for KIT-driven melanoma tumor formation have been studied at the molecular level using isogenically matched and phenotypically defined cells.

Background: Methicillin-resistant Staphylococcus aureus (MRSA) has been an important nosocomial pathogen in our neonatal units since 1990s. To understand the longitudinal changing molecular epidemiology of these MRSA isolates, we conducted this study. Materials: From 2003 to 2018, we collected clinical MRSA isolates from 536 infants hospitalized at neonatal units of a medical center in northern Taiwan. First isolate from each infant was characterized. Results: The case/isolate number ranged from 7 cases/isolates (the lowest) in 2010 to 71 cases/isolates (the highest) in 2004. Of the 536 isolates, a total of 15 pulsotypes were identified. Three major clones were identified and characterized as sequence type (ST) 239/pulsotype A/staphylococcal chromosomal cassette (SCC) mec III/Panton-Valentine leukocidin (PVL)-negative, accounting for 22.2% of the isolates, ST59/pulsotype C/SCCmec IV/PVL-negative, accounting for 34.3% and ST59/pulsotype D/SCCmec VT/PVL-positive, accounting for 30.0%. The first clone (hospital strains) dominated in the first two years, and became weakened from 2005 through 2016. Clonal complex (CC) 59 (combined the second and third clones) dominated (>50% of the isolates) from 2005 through 2018. One community clone (ST573) demonstrated a marked increase since 2007 and vanished abruptly since 2010. Several minor MRSA clones emerged after 2010. Conclusion: The molecular epidemiology of MRSA isolates in our neonatal units from 2003 to 2018 revealed that an epidemic as well as endemic hospital clone of ST239 dominated before 2005 and was replaced by the local community clone of CC59 thereafter.

Aedes aegypti (Culicidae), the mosquito that causes yellow fever, is regarded as a significant vector for many disease agents. The current study sought to learn more about mosquito mid-gut bacteria and their impacts. Mosquito specimens were gathered in Jeddah city (Saudi Arabia), and their gut bacterial flora was then analyzed. The collected mosquitoes exhibit all the characteristic features of A. aegypti, according to a microscopic study. Utilizing the partial mitochondrial cytochrome oxidase (COI) gene analysis, molecular testing established the identity of A. aegypti as a close relative of previously recognized Aedes species submitted in the GenBank, particularly those from Kenya. Unfortunately, based on the results of RT-PCR, none of the Aedes mosquitoes under study had dengue virus (DENV) present. On nutrient agar plates, the mid-gut bacteria were isolated. A total of 34 Gram-positive bacteria were isolated and identified at the molecular level using the 16S rRNA gene and were divided into the two genera Bacillus and Lysinibacillus within the family Bacillaceae. To our knowledge, this study is the first to examine the normal existence of bacterial flora in the gut of DENV-free mosquitos in Saudi Arabia. It is advised to conduct more research to determine how these bacteria affect the transmission of harmful pathogens carried by mosquitoes. Additionally, further research into the antibacterial and anticancer activities of metabolites extracted from the mid-gut bacteria may help in the development of unique drugs.

Goose haemorrhagic polyomavirus (GHPV) is the viral agent of hemorrhagic nephritis and enteritis of geese (HNEG), a lethal disease of goose. The study describes the results of a molecular analysis Polish isolates of GHPV from geese and free-living birds based on complete VP1 gene and VP2 gene sequences. The sequences were analyzed and aligned with different GHPV isolates sequences accessible in the GenBank database. This study indicates affiliation GHPV isolates from fee-living birds and GHPV isolates circulating in Polish goose flocks and around the world to the same genetic groups, which proves their evolutionary relationship and indicates the potential role of free-living birds as a source of infections for poultry.

Ligusticum chuanxiong Hort. as an important Chinese medicinal herb clinically used as anti-inflammatory, antioxidant, and hepatoprotective agents, is widely planted in China. However, related studies on L. chuanxiong’s distribution and significant environmental factors that affect its growth are insufficient. Based on climatic, topographic and soil factors, this study predicted current and future distributions of L. chuanxiong and analyzed the distribution transformation under different scenarios. Moreover, the most important environmental factors for modeling were explored using maximum entropy models, chemical analysis and molecular analysis. Results suggested that the predicted distribution of L. chuanxiong was wider than previously reported. Among these environmental variables, climate factors, especially the minimum temperature of the coldest month (Bio6, 46.7%) and solar radiation (SRAD, 43.4%) contributed more than others to L. chuanxiong’s distribution with optimum values of 0–1.5 °C and 5000–11,000 kJ/m2 per day. Total and highly suitable areas respectively increased by 26,788–943,820 km2 and 34,757–340,417 km2 in the future (2061–2080, 2081–2100). The distribution centers of suitable zones were predicted to migrate north in the future, and the migration distance was 135.74–479.77 km from current center. Results of chemical content determination suggested that L. chuanxiong should be cultivated in high-suitable places to improve medicinal quality by evaluating contents of ferulic acids and Z-ligustilide. Correlation analysis suggested that both chemical contents and gene expression levels decreased with decreasing habitat suitability, suggesting a strong link between environments, chemical constituents, and gene expression. These findings improve the comprehension of the effects of environments on the distribution patterns of L. chuanxiong, as well the relation between environmental suitability and medicinal quality. These findings provide a useful foundation for the planting, cultivation and conservation of L. chuanxiong.

Present study was aimed at screening and characterizing thermostable amylase-producing bacteria from water and sediment samples of unexplored hot spring of Tatta Pani Kotli Azad Kashmir. Four thermophilic isolates were characterized on morphological, biochemical, physiological basis and were authenticated by molecular analysis. By 16S rDNA sequencing, isolates were identified as Anoxybacillus mongoliensis (MBT001), Anoxybacillus flavithermus (MBT002), Bacillus (MBT004). Among all identified strains, MBT003 showed maximum homology with both Anoxybacillus mongoliensis and Anoxybacillus flavithermus. Amylase activity was analyzed qualitatively in starch agar and quantitatively by DNS method. The optimal enzyme production was observed and authenticated by Response Surface Methodology at 7 pH, 70 °C, 1.25% substrate concentration, 300 μL of inocula volume after 48 h of incubation. Optimum amylase activity (4.4 U/mL) and stability (3.3 U/mL) was observed with 1.5% soluble starch at 70 °C. Maximum activity (3.7 U/mL) and stability (1.5 U/mL) was found at pH 8. Enzyme activity was increased in the presence of MgSO4 and CaCl2. Amylase was stable with surfactants and commercial detergents for 30 min. Supplementation of the enzyme with commercial detergent improved the washing ability of the detergent. This investigation has revealed that these thermostable bacteria are excellent source of amylase which can be used commercially for generating economic activity on sustainable basis.

The rising understanding of the programmed death ligand 1 (PD-L1) expression and its molecular foundation has served as an innovating force in both diagnostic assessment and therapy of lung cancer. However, we notice a trend regarding vast heterogeneity of PD-L1 expression, mutation, and further genetic characteristics.We report a unique case of a patient with four synchronous adenocarcinomas with variations in the genetic expression. In case of multiple tumor nodules, adjuvant chemotherapy is discussed in most of the cases. In our patient, the identification of synchronous lung cancer made us pursue a “wait and watch” therapy, rather than chemotherapy. Clinical follow up as well as staging show complete remission without any signs of new tumor manifestations. Therefore, we conclude that it is necessary to clearly understand the genetic foundation of synchronous lung cancer as well as it is variations to offer the best possible treatment for this complex patient group.

The cattle egret (B. ibis) as a common wader birds in Egypt, they act a sole reservoir for many parasites and play a vital role in their life cycle and their distribution in their environment. The study was conducted from September 2020 to August 2021. A total of 180 B. ibis were collected from Al Qantara Gharb, Ismailia province, Egypt. Parasite species identification infecting cattle egret included morphological and morphometric characteristics based on light and scanning microscopy. Additionally, utilizing the partial small subunit ribosomal RNA (18S rRNA, ITS2 and ITS1) gene sequence, maximum parsimony was used to infer the phylogeny of the recovered species. The morphological and molecular studies revealed three helminths. Only one nematode (Desportesius invaginatus, linstow,1901) and two trematodes (Patagifer bilobus, Dietz,1909 and Apharyngostrigea cornu, Zeder,1800) have been identified. The cattle egret (B. ibis) are protagonists in the life cycle of many parasites. The study is considered the first in Egypt to fill the gap of phylogenetic analysis of three helminths; two of them (A. cornu and P. bilobus) were the first to be molecular phylogenetically analyzed in Egypt. The molecular data provided set the conspecific relation of the three identified helminths species with other related helminths. The new identified sequences will help in founding the basis for forthcoming identifications of other helminths species from cattle egret in Egypt and prospective view to possible parasitic assemblage affecting egret population and other animal populations in their environment., Competing Interests: Declaration of competing interest Not Applicable., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The Arabian Peninsula is home to many unique organisms due to its distinctive geographical location. Seven species of amphibians inhabit Saudi Arabia. One of them is the Dhofar toad (Bufo dhufarensis), a species of toad in the family Bufonidae found in Saudi Arabia, the United Arab Emirates, Yemen and Oman. This species lives near freshwater springs, pools, dams, rivers and rural gardens. By evaluating the genetic variety within evolutionary lineages using sequence analysis, we were able to determine the degree of genetic diversity of the Dhofar toad (Bufo dhufarensis). Additionally, we examine the probable connections between this species and other toads and frogs. We collected 27 samples from Bufo dhufarensis and prepared the samples for DNA extraction and PCR using suitable primers. The 16S ribosomal RNA (rRNA) gene sequences in toads from Riyadh Province were determined for the first time and deposited in a public gene data repository. Bufo dhufarensis, which was found in Riyadh Province, is very closely related to the toad Bufo dhufarensis, which was found in Oman, according to a phylogenetic tree based on the 16S rRNA sequence. This is the first report of the genetic diversity of Bufo dhufarensis in Riyadh Province based on the 16S rRNA gene.

Both reticuloendotheliosis and Marek's disease are neoplastic diseases of chickens caused by reticuloendotheliosis virus (REV) and Marek's disease virus (MDV), respectively. The infection of REV or MDV may lead to clinical tumors and also result in immunosuppression and easily allow secondary infection by other pathogens. Here, we investigated a breeder flock of three-yellow chickens in southern China that had been vaccinated with CVI988/Rispens at hatching and had experienced depression, weakness, reduction in weight gain, and an increased death rate after 120 d of age. The morbidity and mortality were 20% and 10%, respectively, at 140 d of age when this infection was diagnosed. The necropsy of the birds revealed significant tumor-like lesions in the heart, liver, spleen, and ceca. Peripheral blood lymphocytes and tumor-like tissues were sampled for PCR detection and for histopathological observation, for virus isolation and the subsequent immunofluorescent assay on the cell cultures and for gene sequencing of the isolated viruses. A REV isolate GX18NNR1 and a MDV isolate GX18NNM5 were both recovered from the sampled bird. Further phylogenetic analysis based on the env gene of REV and the meq gene of MDV demonstrated that GX18NNR1 was closely related to the reference REV strain MD-2, which was isolated from a contaminated commercial turkey herpesvirus vaccine. In addition, the GX18NNM5 was found to belong to the Chinese very virulent MDV strains' cluster. The coinfection of REV and MDV may contribute to tumor outbreaks with high morbidity and mortality in three-yellow chicken flocks.

The cardiopulmonary nematodes Angiostrongylus vasorum, Crenosoma vulpis, Capillaria aerophila and Aelurostrongylus abstrusus, are a cause of concern in the scientific and veterinary community, potentially causing significant disease in domestic animals. To investigate the potential of wild carnivores as reservoir hosts to these parasites, a total of 1041 animals from seven regions of Denmark were sampled: 476 raccoon dogs (Nyctereutes procyonoides), 367 red foxes (Vulpes vulpes), 123 American mink (Neovison vison), 31 beech martens (Martes foina), 30 Eurasian otters (Lutra lutra) and 14 polecats (Mustela putorius). Hearts and lungs were collected and examined for cardiopulmonary parasites. Capillaria aerophila was identified using morphology, whereas A. vasorum and C. vulpis were identified by a duplex real-time PCR, and A. abstrusus by conventional PCR. This is the first Danish report of A. vasorum and C. vulpis infections in raccoon dogs, mink and polecats, and of C. aerophila in raccoon dogs and beech martens. In addition, this is the first time A. vasorum and C. vulpis have been identified in wild animals from the island of Bornholm, just as it is the first report of C. vulpis in American mink, and C. vulpis and A. vasorum in polecats in Europe. The prevalence of A. vasorum appears to have increased in red foxes in Denmark compared to previous studies, while C. vulpis and C. aerophila prevalences are lower. Our data show that several wild carnivores can serve as reservoir hosts for A. vasorum, C. vulpis and C. aerophila in Denmark, and that A. vasorum appears more abundant than previously reported. It is speculated that the A. vasorum increase might relate to increased snail abundance, which may be due to a rise in mean yearly temperatures in Denmark.

Cyclops spp. is among the most common zooplankton in fish larval rearing tanks of the Marine Hatchery of the National Institute of Oceanography and Fisheries, Alexandria, Egypt. This study aims to investigate the molecular analysis of this species and to evaluate the effect of a feeding diet composed of soybean, yeast, and microalgae (Tetraselmis chuii) on their population density and nutritional value. Phylogenetic identification of this species was conducted using (12S) rRNA molecular marker technique. The analysis showed that the investigated sample could be identified as Cyclops abyssorum divergens. Different concentrations of soybean and yeast ranging between 0.1 and 0.5 mg.l−1, and different microalgal density population ranging between 1.0 and 5 × 106 cells.ml−1 were used to feed cyclops for six consecutive days. The results showed that the treatment fed with soybean (0.5 µg/ml) recorded the highest population density of Cyclops (was 27 individuals/ml). As for the nutritional value, C. abyssorum divergens fed on T. chuii recorded the highest protein, lipids, and total amino acid values. Moreover, the highest value of eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) and other polyunsaturated fatty acids (PUFA) were detected in the C. abyssorum divergens growth fed on T. chuii.

Background: The new diagnostic concept of liquid biopsy is based on the analysis of circulating tumor cells (CTCs) and cell-free DNA (ctDNA). In addition to providing a more comprehensive view of the tumor characteristics including molecular variations, ctDNA analysis through liquid biopsies may also allow for a non-invasive, rapid, and cost-effective identification of biomarkers for tumor detection and monitoring of tumor progression. Objective: In this review, we summarize key active clinical trial studies involving utilization of ctDNA derived from liquid biopsy in the early and metastatic breast cancer setting. With this, we also provide a brief overview of the potential future implementations of the LB technology and outlining how ctDNA analysis needs to be standardized through the performance of similar clinical studies. Methods: A review was conducted on Clinicaltrials.gov to identify active trials related to use of circulating tumor DNA (ctDNA) for breast cancer. Search terms included “breast cancer,” “liquid biopsy,” and “ctDNA.” Conclusion: While LB is gaining traction in many cancer settings, its use in BC is still early and warrants more investigation in breast cancer diagnostic and treatment settings, including early detection of disease recurrence.

The organic matter molecular mechanism by which combined hydrothermal carbonization (co-HTC) of municipal sludge (MS) and agricultural wastes (rice husk, spent mushroom substrate, and wheat straw) reduces the inhibitory effects of aqueous phase (AP) products on pak choi (Brassica campestris L.) growth compared to HTC of MS alone is not clear. Fourier-transform ion cyclotron resonance mass spectrometry was used to characterize the differences in organic matter at the molecular level between AP from MS HTC alone (AP-MS) and AP from co-HTC of MS and agricultural waste (co-Aps). The results showed that N-bearing molecules of AP-MS and co-Aps account for 70.6 % and 54.2 %-64.1 % of all molecules, respectively. Lignins were present in the highest proportion (56.3 %-78.5 %) in all APs, followed by proteins and lipids. The dry weight of co-APs hydroponically grown pak choi was 31.6 %-47.6 % higher than that of the AP-MS. Molecules that were poorly saturated and with low aromaticity were preferentially consumed during hydroponic treatment. Molecules present before and after hydroponics were defined as resistant molecules; molecules present before hydroponics but absent after hydroponics were defined as removed molecules; and molecules absent before hydroponics but present after hydroponics were defined as produced molecules. Large lignin molecules were broken down into more unsaturated molecules, but lignins were the most commonly resistant, removed, and produced molecules. Correlation analysis revealed that N- or S-bearing molecules were phytotoxic in the AP. Tannins positively influenced the growth of pak choi. These results provide new insights into potential implementation strategies for liquid fertilizers produced from AP arising from HTC of MS and agricultural wastes., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

The chewing louse genus Eutrichophilus Mjöberg has 19 species only associated with porcupines (Rodentia: Erethizontidae). Of these species, E. cercolabes, E. cordiceps, E. emersoni, E. minor, E. moojeni, and E. paraguayensis have been recorded in Brazil. In the present study, we report E. cordiceps for the first time in the São Paulo State (Bauru Municipality) and for the second time in the Santa Catarina State (Lages Municipality), providing scanning electron images and light microscopy for the eggs, as well as the first molecular data (18S rRNA) for the genus. Additionally, Bartonella sp. was detected for the first time in this chewing lice species., Competing Interests: Declaration of competing interest The authors declare that they have no conflict of interest., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Background: Feline infectious peritonitis (FIP) is a fatal immune-mediated disease in cat, caused by mutated feline coronavirus (FCoV). Due to its difficulties in diagnosis, FIP is sometimes underdiagnosed. Therefore, several laboratory procedures were performed to gain high index suspicion of FIP. However, through several laboratory findings, not only FIP but also SEZ infection was confirmed in this case. Case description: A-year-old male, domestic cat was admitted to Veterinary Medicine Clinical Pathology Laboratory, Universitas Gadjah Mada, for further effusion examination due to its high suspicion of feline infectious peritonitis (FIP). Further examination using molecular and post-mortem analysis resulted on confirmed SEZ infection and FIP. This study informed the manifestation and pathological changes in patient with SEZ and FIP in the same time. Conclusions: This study showed that viral infection followed by bacterial infection could be fatal and untreatable. After these findings, clinicians may consider SEZ infection in cat with respiratory disorder followed by thoracic effusion besides FIP. Companion animal, especially outdoor-kept animal, possibly become infected from its contact to another human or animal in the environment.

Fowl adenovirus strains were isolated from the internal organs of 3-wk-old broiler flocks exhibited clinical signs associated with inclusion body hepatitis (IBH). The isolated strains were molecularly characterised and sequencing revealed three distinct clusters. One cluster showed close proximity at the nucleotide level with adenovirus type/species - 6/E, 7/E, 8a/E, and 8b/E. The second cluster contained five reference sequences belonging to the species FAdV-D and E. A third cluster contained one field and four reference sequences belonging to the FAdV-5/B, FAdV-4/C, FAdV-2/D, and FAdV-1/A type/species respectively. The heterogenicity, Relative Synonymous Codon Usage (RSCU), codon composition, and nucleotide frequencies were examined. Statistical analyses, were carried out. The maximum likelihoods for the examined sequences were estimated. The data indicated that correlation between isolated of adenovirus type/species 5/B, and E in Poland have been presented. Indicated adenovirus types and their combinations with locally circulating FAdVs strains could have implications for current detection methods and pathogenicity on infected chickens.

Background: Classical galactosemia is an autosomal recessive inherited metabolic disorder caused by mutations in the galactose-1-phosphate uridyltransferase (GALT) gene. GALT enzyme deficiency leads to the accumulation of galactose-1-phosphate in various organs, causing hepatic, renal and cerebral impairment. Over 300 mutations have been reported in the GALT gene. The aim of this study was to describe molecular characterization of GALT gene in Argentinian patients with decreased GALT activity, and to correlate molecular results with enzyme activity. Methods: 37 patients with enzyme activity below 6.3 μmol/h/g Hb (35% of normal value) were included. GALT activity was measured on red blood cells. DNA was extracted from peripheral blood. p.Gln188Arg mutation was studied by PCR-RFLP and, on samples negative or heterozygous, GALT gene was sequenced. In vivo splicing analysis of the GALT gene was performed on RNA extracted from leukocytes of one patient. Results: 14 different sequence variations were identified among 72 unrelated alleles. The two most common disease-causing mutations were p.Gln188Arg (24/72) and p.Lys285Asn (9/72). Three novel mutations were detected. One of them, c.688G>A, caused partial skipping of exon 9 of the GALT gene. Enzyme activity correlated with GALT genotype in 36 of the 37 patients. Conclusion: This is the first report of sequence variations in the GALT gene in the Argentinian population. This study highlights the contribution of the molecular analysis to the diagnosis of Galactosemia and reveals c.688G>A as a novel Duarte-like variant, with a high prevalence in our population.

Cyanobacterial harmful algal blooms (CyanoHABs) threaten public health and freshwater ecosystems worldwide. In this study, our main goal was to explore the dynamics of cyanobacterial blooms and how microcystins (MCs) move from the Lalla Takerkoust reservoir to the nearby farms. We used Landsat imagery, molecular analysis, collecting and analyzing physicochemical data, and assessing toxins using HPLC. Our investigation identified two cyanobacterial species responsible for the blooms: Microcystis sp. and Synechococcus sp. Our Microcystis strain produced three MC variants (MC-RR, MC-YR, and MC-LR), with MC-RR exhibiting the highest concentrations in dissolved and intracellular toxins. In contrast, our Synechococcus strain did not produce any detectable toxins. To validate our Normalized Difference Vegetation Index (NDVI) results, we utilized limnological data, including algal cell counts, and quantified MCs in freeze-dried Microcystis bloom samples collected from the reservoir. Our study revealed patterns and trends in cyanobacterial proliferation in the reservoir over 30 years and presented a historical map of the area of cyanobacterial infestation using the NDVI method. The study found that MC-LR accumulates near the water surface due to the buoyancy of Microcystis. The maximum concentration of MC-LR in the reservoir water was 160 µg L -1 . In contrast, 4 km downstream of the reservoir, the concentration decreased by a factor of 5.39 to 29.63 µgL -1 , indicating a decrease in MC-LR concentration with increasing distance from the bloom source. Similarly, the MC-YR concentration decreased by a factor of 2.98 for the same distance. Interestingly, the MC distribution varied with depth, with MC-LR dominating at the water surface and MC-YR at the reservoir outlet at a water depth of 10 m. Our findings highlight the impact of nutrient concentrations, environmental factors, and transfer processes on bloom dynamics and MC distribution. We emphasize the need for effective management strategies to minimize toxin transfer and ensure public health and safety., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier B.V. All rights reserved.)

Objectives: The identification of the specific pathogen responsible for a respiratory infection in patients with hematological malignancies (HM) would ensure relevant treatment and prevent toxicity associated with anti-infective therapy. This large-scale study aimed to explore the clinical impact of fiberoptic bronchoscopy with bronchoalveolar lavage (FOB-BAL) in conjunction with molecular analysis on the diagnosis and management of respiratory infections in hemato-oncological patients. Methods: All consecutive patients with HM and pulmonary infiltrates, who underwent FOB-BAL between January 2008 and January 2013, were included in the analysis. Clinical characteristics, FOB-BAL results, and treatment adjustments were recorded, and factors predicting a positive BAL were assessed. Results: Four hundred and twenty-five FOB-BAL procedures were analyzed. BAL revealed a specific diagnosis in 219 (51.5%) patients, 208 of them with a pulmonary infection. Infectious etiological agents found were mainly Aspergillus spp (n = 142), bacterial species (n = 44), and Pneumocystis jirovecii (n = 34). Multivariate analysis showed that a lymphoproliferative disease, ≥2 symptoms (dyspnea/cough/hemoptysis/pleuritic pain), and less than 4 days between symptom appearance and FOB-BAL, predicted a positive FOB-BAL result. BAL results prompted a treatment modification in 48% of subjects. Conclusions: FOB-BAL in conjunction with molecular assays is efficient in the rapid detection of life-threatening infections, allowing for adjustment of anti-infective therapy, which may result in better outcomes and reduce treatment-related toxicity.

The present study provides a comprehensive assessment of the quantitative analysis by high-performance liquid chromatography coupled with dual orthogonal electrospray ionization time-of-flight mass spectrometry (HPLC-ESI-TOF-MS) of pinene markers, biomass-burning related phenols, and other relevant carboxylic acids in atmospheric aerosol samples. Significant insights into the quantitative determination are offered on the basis of systematic experiments targeting the optimization of chromatographic separation, ionization source, and mass spectrometer performance. After testing three analytical columns, the best separation of the compounds of interest was achieved on a Poroshell 120 ECC18 column (4.6 × 50 mm, 2.7 µm) thermostated at 35 °C, operating in gradient elution mode with 0.1% acetic acid in water and acetonitrile at a 0.8 mL min -1 flow rate. Optimal operational conditions for the ESI-TOF-MS instrument were identified as a 350 °C drying gas temperature, 13 L min -1 drying gas flow rate, 60 psig nebulizer pressure, 3000 V for the ion transfer capillary, 60 V for the skimmer, and 150 V for the fragmentor. Additionally, the matrix effect on the ESI efficiency and the spike recovery factors of the compounds were tested. Method quantification limits can go as low as in the 0.88-48.0 μg L  -   1 (3.67-200 pg m  -   3 , at 120 m 3 of sampled air) range. The developed method was shown to be reliable for the quantification of the targeted compounds in real atmospheric aerosol samples. The accuracy in the molecular mass determination of less than 5 ppm and the acquisition in the full scan mode were shown to bring additional insights into the organic constituents in atmospheric aerosols., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Introduction: Mantle cell lymphoma (MCL) is a rare type of lymphoma, which despite improvements in therapies in recent decades, remains an incurable disease. There is currently no reliable marker of chemoresistance available. In this study, we investigated the prognostic role of MIPIb and the association with biological markers including SOX11, p53 expression, Ki-67, and CDKN2A., Materials and Methods: This retrospective study was focused on 23 patients with newly diagnosed classical MCL, treated at the University Hospital of Bari (Italy) between January 2006 and June 2019., Results: We identified a MIPIb value ≥ 5.4440 as a prognostic parameter that correlates with p53 expression and CDKN2A deletion. We also observed that patients with p53 overexpression had a significantly higher MIPIb (5.52 ± 0.53) which in 80% of patients had a value higher than 5.4440. On the other hand, CDKN2A deletion was found more frequently (75%) associated with MIPIb ≥5.4440. Only the CDKN2A deletion was associated with a higher proliferation index, with 66.7% of samples having Ki67 ≥30%. From the survival analysis we found that patients with p53 overexpression and CDKN2A deletion have a significantly worse prognosis with a median overall survival of 50 (P = .012) and 52 months (P = .018), respectively., Conclusion: p53 expression and CDKN2A deletion represent a reliable pretreatment prognostic factor that identifies patients who do not benefit from currently used immunochemotherapy-based therapies and who are candidates for diversified treatments with the aim of improving prognosis. The MIPIb represents a prognostic index that correlates well with these biological alterations and can be used in clinical practice as their surrogate., Competing Interests: Disclosure The authors declare no conflict of interest., (Copyright © 2023 Elsevier Inc. All rights reserved.)

Characterization of atmospheric fine particulate matter (PM 2.5 ) in large cities has important implications for the study of their sources and formation mechanisms, as well as in developing effective measures to control air pollution. Herein, we report a holistic physical and chemical characterization of PM 2.5 by combining surface-enhanced Raman scattering (SERS) with scanning electron microscopy (SEM) and electron-induced X-ray spectroscopy (EDX). PM 2.5 particles were collected in a suburban area of Chengdu, a large city in China with a population over 21 million. A special SERS chip composed of inverted hollow Au cone (IHAC) arrays was designed and fabricated to allow direct loading of PM 2.5 particles. SERS and EDX were used to reveal the chemical composition, and particle morphologies were analyzed from SEM images. SERS data of atmospheric PM 2.5 indicated qualitatively the presence of carbonaceous particulate matter, sulfate, nitrate, metal oxides and bioparticles. The EDX showed the presence of the elements C, N, O, Fe, Na, Mg, Al, Si, S, K, and Ca in the collected PM 2.5 . Morphology analysis showed that the particulates were mainly in the form of flocculent clusters, spherical, regular crystal shaped or irregularly shaped particles. Our chemical and physical analyses also revealed that the main sources of PM 2.5 are automobile exhaust, secondary pollution caused by photochemical reactions in the air, dust, emission from nearby industrial exhaust, biological particles, other aggregated particles, and hygroscopic particles. SERS and SEM data collected during three different seasons showed that carbon-containing particles are the principal sources of PM 2.5 . Our study demonstrates that the SERS based technique, when combined with standard physicochemical characterization methods, is a powerful analytical tool to determine the sources of ambient PM 2.5 pollution. Results obtained in this work may be valuable to the prevention and control of PM 2.5 pollution in air., Competing Interests: Declaration of competing interest The authors declared no potential conflicts of interest with respect to the research, authorship, and/or publication of this article., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Management of high grade, serous and/or endometrioid, advanced (stages III-IV) ovarian carcinomas and HRD-BRCA testing in 2023: update according to data published/presented in 2022 Molecular analysis of ovarian carcinomas must be now systematically performed to determine BRCA1 and BRCA2 status as well as genomic instability score. Several types of tests are available. From a clinical perspective, new data from phase III clinical trials presented in 2022 confirm the key role of PARP inhibitors in first-line medical treatment of high-grade serous ovarian cancers. A new algorithm that includes all new evidence is proposed for selection of first-line therapy., (Copyright © 2023 Elsevier Masson SAS. Tous droits réservés. All rights reserved.)

Basidiomata of a wild mushroom macroscopically recognised as Pleurotus ostreatus were observed on an oak trunk in a mixed wood of northern Albania. Pure cultures of the fungus were then obtained on potato-dextrose-agar medium. Molecular analyses of genomic DNA of the fungus confirmed its identification. The rDNA ITS region nucleotide sequence of the studied Pleurotacea matched at 99% those of two P. ostreatus strains already present in NCBI GenBank database. The rDNA ITS nucelotide sequences of two pure cultures of the Albanian P. ostreatus were deposited in EMBL database under the accession numbers LN849458 and LN849459. One of the fungus isolates was subsequently cultivated under protected and semi-natural conditions. Productivity and biological efficiency of the Albanian P. ostreatus ranged from about 10% to 16% and from 33 to 53.33%, respectively. This seems to be the first report on the artificial cultivation of P. ostreatus in Albania and could have, in the next future, a high economic impact on development and diffusion of this important edible mushroom over the country.

Israel conducts routine environmental (15 sites) and acute flaccid paralysis (AFP) surveillance for poliovirus. During September 2021, increasing numbers of wastewater samples collected from more than one site in the Jerusalem region proved positive for ambiguous type 3 vaccine-derived poliovirus (aVDPV3), while environmental samples from remaining sampling sites were negative. In late February 2022, a VDPV3, genetically related to the Jerusalem environmental surveillance samples, was isolated from a stool sample collected from a non-immunodeficient, non-immunized child from Jerusalem who developed AFP, indicating that the aVDPV3s were circulating (cVDPV3s) rather than immunodeficiency-related VDPV3s (iVDPVs). In response to these isolations, the Israel Ministry of Health launched a catch-up immunization program., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Background: Pleural biopsy findings offer greater diagnostic sensitivity in malignant pleural effusions compared with pleural fluid. The adequacy of pleural biopsy techniques in achieving molecular marker status has not been studied, and such information (termed "actionable" histology) is critical in providing a rational, efficient, and evidence-based approach to diagnostic investigation., Research Question: What is the adequacy of various pleural biopsy techniques at providing adequate molecular diagnostic information to guide treatment in malignant pleural effusions?, Study Design and Methods: This study analyzed anonymized data on 183 patients from four sites across three countries in whom pleural biopsy results had confirmed a malignant diagnosis and molecular profiling was relevant for the diagnosed cancer type. The primary outcome measure was adequacy of pleural biopsy for achieving molecular marker status. Secondary outcomes included clinical factors predictive of achieving a molecular diagnosis., Results: The median age of patients was 71 years (interquartile range, 63-78 years), with 92 of 183 (50%) male. Of the 183 procedures, 105 (57%) were local anesthetic thoracoscopies (LAT), 12 (7%) were CT scan guided, and 66 (36%) were ultrasound guided. Successful molecular marker analysis was associated with mode of biopsy, with LAT having the highest yield and ultrasound-guided biopsy the lowest (LAT vs CT scan guided vs ultrasound guided: LAT yield, 95%; CT scan guided, 86%; and ultrasound guided, 77% [P = .004]). Biopsy technique and size of biopsy sample were independently associated with successful molecular marker analysis. LAT had an adjusted OR for successful diagnosis of 30.16 (95% CI, 3.15-288.56; P = .003) and biopsy sample size an OR of 1.18 (95% CI, 1.02-1.37) per millimeter increase in tissue sample size (P < .03)., Interpretation: Although previous studies have shown comparable overall diagnostic yields, in the modern era of targeted therapies, this study found that LAT offers far superior results to image-guided techniques at achieving molecular profiling and remains the optimal diagnostic tool., (Copyright © 2022 American College of Chest Physicians. Published by Elsevier Inc. All rights reserved.)

In the last decade, non-small-cell lung cancer (NSCLC) treatment has improved with the approval of multiple therapies to target specific genetic alterations. Though, next generation sequencing (NGS) has traditionally been conducted from tissue biopsy samples, developing data supports the use of plasma-based circulating tumor DNA (ctDNA), also known as "liquid biopsy," to complement tissue biopsy approaches in guiding front-line therapy. This study is a retrospective analysis of 170 new NSCLC patients treated at 2 cancer centers within a 5-year period who received both tissue and liquid biopsy NGS as standard of care. Based on a treatment schema defined by testing sufficiency, biomarker detection, and turnaround time (TAT), physicians based the majority of their treatments on liquid biopsy results (73.5%) versus tissue biopsy (25.9%). Liquid biopsy NGS returned results on average 26.8 days faster than tissue and reported higher testing success. For guideline-recommended biomarkers, liquid biopsy was 94.8% to 100% concordant with tissue. In comparing testing modalities, a liquid-first approach identified guideline-recommended biomarkers in 76.5% of patients versus 54.9% in a tissue-first approach. There was no significant difference in time-to-treatment, or survival outcomes (overall survival and progression free survival) based on liquid versus tissue biopsy findings. This research demonstrates that liquid biopsy NGS is an effective tool to capture actionable genetic alterations in NSCLC. Due to its high concordance to tissue, faster TAT, and similarity in outcomes and time-to-treatment, liquid biopsy can be used either as a first-line test or concordantly with tissue biopsy to guide treatment decisions in NSCLC., Competing Interests: Disclosure LR reports research support from: Guardant Health, Nant Health, Exosomes DX, Liquid Genomics, BMS, Merck, Genentech, Astra-Zeneca, Lilly Oncology, LOXO and Natera. ES: Speaker and Honoraria from Guardant Health, Astra-Zeneca, Merck, BMS, Lilly Oncology, Takeda, Regeneron., (Copyright © 2022 Elsevier Inc. All rights reserved.)

Introduction/background: Samples from endobronchial ultrasound-guided fine needle aspiration (EBUS-TBNA) are frequently used for next generation sequencing (NGS) in patients with non-small cell lung cancer (NSCLC) to look for genetic driver mutations. The objective of the current study was to evaluate the performance of extended NGS panels using EBUS-TBNA samples in a real-world setting and identify factors associated with the success of NGS., Materials and Methods: This study included all patients who underwent EBUS and were diagnosed with non-squamous NSCLC with mediastinal metastasis from 2016 to 2019 at the University of Pennsylvania. We reviewed demographic information, imaging studies, procedure reports, pathology and NGS reports. Logistic regression was used to analyze factors associated with the success of NGS panels., Results: The success rates of NGS using EBUS-TBNA samples were 92.5%, and 91.5% for DNA and RNA NGS panels respectively. Samples from higher N stage (N2 and N3 lymph nodes) and with higher tumor cellularity (>25%) resulted in higher success rate for DNA NGS. The effect of tumor cellularity remained borderline significant after entering multivariable logistic regression. The short-axis diameter of the sampled lymph node on CT scan, FDG-avidity on PET CT and >3 EBUS passes per lymph node during the procedure were not associated with NGS success., Conclusion: Both DNA and RNA extended-panel NGS had high performance using EBUS-TBNA samples. Sampling more advanced nodal stations and obtaining samples with higher tumor cellularity were associated with higher success rate of DNA NGS. Other imaging or procedural factors did not affect NGS performance., (Copyright © 2022 Elsevier Inc. All rights reserved.)

A leucemia megacariocítica aguda (LMA-M7) é um subtipo raro de leucemia mielóide aguda (LMA) que foi recentemente incorporada na classificação Franco-Americana-Britânica (FAB). Ela representa 3% a 5% dos casos de LMA, sendo freqüentemente associada a mielofibrose e retrata um subtipo de mau prognóstico. O diagnóstico da LMA-M7 baseia-se, inicialmente, nas características morfológicas das células leucêmicas. O aspirado de medula óssea, ou biópsia, mostra uma população de células pleomórficas e basofílicas, podendo apresentar projeções citoplasmáticas (blebs). A utilização apenas de critérios morfológicos e citoquímicos não é suficiente para um diagnóstico correto, por isso faz-se necessária uma diferenciação de leucemia megacariocítica aguda com os outros subtipos de leucemia mielóide aguda, principalmente nos casos em que as células blásticas se apresentam indiferenciadas, como é o caso da leucemia mielóide aguda, minimamente diferenciada (LMA-M0), da leucemia mielóide aguda sem maturação (LMA-M1) e da leucemia linfóide aguda, subtipo L1 e L2. Sendo assim, a utilização de técnicas de imunofenotipagem é essencial para o diagnóstico diferencial, pois mostra uma população de células leucêmicas com ausência da maioria dos marcadores linfóides e mielóides de superfície, mas com expressão para os antígenos da linhagem megacariocítica: CD41a (complexo glicoproteíco IIb/IIIa), CD42b (glicoproteína Ib) e/ou CD61 (glicoproteína IIIa), permitindo uma classificação correta em 98% dos casos. Acrescenta-se também o estudo das anormalidades cromossômicas identificadas por técnicas de citogenética e análise molecular que passa a ser importante para a determinação do prognóstico e definição do regime terapêutico.Acute megakaryocytic leukemia (AML-M7) is a rare subtype of acute myeloid leukemia (AML), which has recently been incorporated in the FAB (French-American-British) classification. It represents from 3 to 5% of AML cases, is frequently associated to myelofibrosis and is a subtype with poor prognosis. The diagnosis of AML-M7 is initially based on the morphologic characteristics of leukemic cells. Bone marrow aspirates or biopsy shows a population of pleomorphic and basophilic cells which may present cytoplasmatic blebs. Morphologic and cytochemical criteria are not enough for a correct diagnosis. It is necessary to differentiate acute megakaryocytic leukemia with other subtypes of acute myeloid leukemia mainly in cases in which blast cells are undifferentiated, as are the cases of AML-M0, AML-M1 and of acute lymphoid leukemia, subtypes L1 and L2. The use of immunopherotypic techniques is essential for a differential diagnosis showing a population of leukemia cells with the absence of most surface lymphoid and myeloid markers, yet relevant for the antigen of the megakaryocytic expression: CD41a (complex glycoproteic IIb/IIIa), CD42b (glycoprotein Ib) and/or CD61 (glycoprotein IIIa), there by providing the for correct classification in 98% of the cases. Chromosomal abnormalities identified by cytogenetic techniques and molecular analysis are important for determining the prognosis and definition of the therapheutic regime.

OBJETIVO: Investigar a prevalência da mutação 35delG em amostra de recém-nascidos, com teste molecular específico; avaliar as perspectivas para a triagem neonatal genética para a deficiência auditiva. CASUÍSTICA E MÉTODO: Foram avaliados 223 recém-nascidos no Hospital de Base de São José do Rio Preto, em São Paulo, para análise molecular da mutação 35delG, no gene da conexina 26, com a técnica da reação em cadeia da polimerase alelo-específico, após extração do DNA genômico de sangue de cordão umbilical. RESULTADOS: Foram identificados cinco heterozigotos, obtendo-se prevalência de 2,24% de portadores da mutação 35delG, na população do estudo. CONCLUSÃO: O uso do teste molecular permitiu a identificação da mutação 35delG na população do estudo, podendo ser utilizado como complemento aos rastreamentos audiométricos neonatais por ser simples, rápido, de fácil execução e de baixo custo.OBJECTIVES: To investigate the prevalence of the 35delG mutation in a newborn population, with specific molecular testing, and to evaluate the prospects for genetic neonatal screening for hearing impairment. POPULATION AND METHOD: 233 newborn were evaluated at the Hospital de Base de São José do Rio Preto, SP, for molecular analysis of the 35delG mutation in the connexin 26 gene, with the reaction technique in allele-specific polymerase chain reaction, after genomic DNA extraction from umbilical cord blood. RESULTS: Five heterozygotes were identified, obtaining a prevalence of 2.24% of 35delG mutation carriers in the study population. CONCLUSION: Using the molecular test allowed for the identification of the 35delG mutation in the study population with the possibility of being used as a complement to neonatal audiometric screening as being simple, fast, and easily to perform with low costs.

The latest European recommendations of the European Societies of Gynecological Oncology (ESGO), Radiotherapy and Oncology (ESTRO) and Anatomopathology (ESP) concerning the management of patients with endometrial cancer were published in 2021. On behalf of the French Society of Gynecologic Oncology (SFOG) and the SFOG campus, we wish to summarize for the French-speaking readership the main measures with a more specific application for France. We also incorporate data from a Delphi survey conducted with a panel of French and French-speaking Swiss experts. The data presented in this article relate to histo-molecular characteristics, radiological data of endometrial cancer, and management of low-risk, intermediate-risk, intermediate-high-risk, and metastatic cancers. The aim of this review article is to show the application of the latest international recommendations to clinicians and pathologists for the implementation of these recommendations., (Copyright © 2022 Société Française du Cancer. Published by Elsevier Masson SAS. All rights reserved.)