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Cadmium (Cd) is a toxic heavy metal that accumulates in plants, negatively affecting their physiological processes, growth, and development, and poses a threat to human health through the food chain. 6-phosphogluconolactonase (PGL) is a key enzyme in the Oxidative Pentose Phosphate Pathway(OPPP) in plant cells, essential for cellular metabolism. The OPPP pathway provides energy and raw materials for organisms and is involved in antioxidant reactions, lipid metabolism, and DNA synthesis. This study describes the Cd responsive gene AetPGL from Aegilops tauschii. Overexpression of AetPGL under Cd stress increased main root length and germination rate in Arabidopsis. Transgenic lines showed higher antioxidant enzyme activities and lower malondialdehyde (MDA) content compared to the wild type. The transgenic Arabidopsis accumulated more Cd in the aboveground part but not in the underground part. Expression levels of AtHMA3, AtNRAMP5, and AtZIP1 in the roots of transgenic plants increased under Cd stress, suggesting AetPGL may enhance Cd transport from root to shoot. Transcriptome analysis revealed enrichment of differentially expressed genes (DEGs) in the plant hormone signal transduction pathway in AetPGL-overexpressing plants. Brassinosteroids (BR), Gibbenellin acid (GA), and Jasmonic acid (JA) contents significantly increased after Cd treatment. These results indicate that AetPGL may enhance Arabidopsis' tolerance to Cd by modulating plant hormone content. In conclusion, AetPGL plays a critical role in improving cadmium tolerance and accumulation and mitigating oxidative stress by regulating plant hormones, providing insights into the molecular mechanisms of plant Cd tolerance. [ABSTRACT FROM AUTHOR]

Simple Summary: The standard interpretation methods of germline variants in cancer are limited due to overlapping features in constitutional and sporadically derived forms of cancers and the unavailability of key differentiating details in public settings. To address this challenge, we established INT2GRATE (INTegrated INTerpretation of GeRmline And Tumor gEnomes), a multi-institution oncology consortium to advance the integrated application of constitutional and tumor data and share the integrated variant data in a publicly accessible knowledgebase. The aim of our study is to introduce INT2GRATE|HPPGL, a platform for the integrated interpretation of hereditary paraganglioma–pheochromocytoma syndromes (HPPGL). We describe the details of the INT2GRATE|HPPGL Variant Evidence Framework for succinate dehydrogenase (SDHx) genes using key HPPGL personal and family history, as well as tumor-derived evidence. We applied the INT2GRATE|HPPGL Variant Evidence Framework to 8600 variants to programmatically process and share the integrated variant data in ClinVar using a custom-made INT2GRATE variant submission pipeline. This novel integrated variant assessment and data sharing in hereditary cancers is essential to help improve the clinical interpretation of genomic variants and advance precision oncology. Standard methods of variant assessment in hereditary cancer susceptibility genes are limited by the lack of availability of key supporting evidence. In cancer, information derived from tumors can serve as a useful source in delineating the tumor behavior and the role of germline variants in tumor progression. We have previously demonstrated the value of integrating tumor and germline findings to comprehensively assess germline variants in hereditary cancer syndromes. Building on this work, herein, we present the development and application of the INT2GRATE|HPPGL platform. INT2GRATE (INTegrated INTerpretation of GeRmline And Tumor gEnomes) is a multi-institution oncology consortium that aims to advance the integrated application of constitutional and tumor data and share the integrated variant information in publicly accessible repositories. The INT2GRATE|HPPGL platform enables automated parsing and integrated assessment of germline, tumor, and genetic findings in hereditary paraganglioma–pheochromocytoma syndromes (HPPGLs). Using INT2GRATE|HPPGL, we analyzed 8600 variants in succinate dehydrogenase (SDHx) genes and their associated clinical evidence. The integrated evidence includes germline variants in SDHx genes; clinical genetics evidence: personal and family history of HPPGL-related tumors; tumor-derived evidence: somatic inactivation of SDHx alleles, KIT and PDGFRA status in gastrointestinal stromal tumors (GISTs), multifocal or extra-adrenal tumors, and metastasis status; and immunohistochemistry staining status for SDHA and SDHB genes. After processing, 8600 variants were submitted programmatically from the INT2GRATE|HPPGL platform to ClinVar via a custom-made INT2GRATE|HPPGL variant submission schema and an application programming interface (API). This novel integrated variant assessment and data sharing in hereditary cancers aims to improve the clinical assessment of genomic variants and advance precision oncology. [ABSTRACT FROM AUTHOR]

Background: Both [18F]FDOPA (FDOPA) and [68Ga]DOTATOC PET/CT (DOTATOC) are widely used for detection of pheochromocytomas/paraganglioma (PPGL). However, direct comparisons of the performance of the two tracers are only available in small series. We conducted a retrospective comparative analysis of FDOPA and DOTATOC to assess their sensitivity and accuracy in detecting PPGL when administered based on suspicion of PPGL. We consecutively included patients referred on suspicion of PPGL or PPGL recurrence who were scanned with both FDOPA and DOTATOC. Both scans were reviewed retrospectively by two experienced observers, who were blinded to the final diagnosis. The assessment was made both visually and quantitatively. The final diagnosis was primarily based on pathology. Results: In total, 113 patients were included (97 suspected of primary PPGL and 16 suspected of recurrence). Of the 97 patients, 51 had pheochromocytomas (PCC) (in total 55 lesions) and 6 had paragangliomas (PGL) (in total 7 lesions). FDOPA detected and correctly localized all 55 PCC, while DOTATOC only detected 25 (sensitivity 100% vs. 49%, p < 0.0001; specificity 95% vs. 98%, p = 1.00). The negative predictive value (100% vs. 63%, p < 0.001) and diagnostic accuracy (98% vs. 70%, p < 0.01) were higher for FDOPA compared to DOTATOC. FDOPA identified 6 of 6 patients with hormone producing PGL, of which one was negative on DOTATOC. Diagnostic performances of FDOPA and DOTATOC were similar in the 16 patients with previous PPGL suspected of recurrence. Conclusions: FDOPA is superior to DOTATOC for localization of PCC. In contrast to DOTATOC, FDOPA also identified all PGL but with a limited number of patient cases. [ABSTRACT FROM AUTHOR]

Abstract Background The structural basis of chloroplast and the regulation of chloroplast biogenesis remain largely unknown in maize. Gene mutations in these pathways have been linked to the abnormal leaf color phenotype observed in some mutants. Large scale structure variants (SVs) are crucial for genome evolution, but few validated SVs have been reported in maize and little is known about their functions though they are abundant in maize genomes. Results In this research, a spontaneous maize mutant, pale green leaf-shandong (pgl-sd), was studied. Genetic analysis showed that the phenotype of pale green leaf was controlled by a recessive Mendel factor mapped to a 156.8-kb interval on the chromosome 1 delineated by molecular markers gy546 and gy548. There were 7 annotated genes in this interval. Reverse transcription quantitative PCR analysis, SV prediction, and de novo assembly of pgl-sd genome revealed that a 137.8-kb deletion, which was verified by Sanger sequencing, might cause the pgl-sd phenotype. This deletion contained 5 annotated genes, three of which, including Zm00001eb031870 , Zm00001eb031890 and Zm00001eb031900, were possibly related to the chloroplast development. Zm00001eb031870, encoding a Degradation of Periplasmic Proteins (Deg) homolog, and Zm00001eb031900, putatively encoding a plastid pyruvate dehydrogenase complex E1 component subunit beta (ptPDC-E1-β), might be the major causative genes for the pgl-sd mutant phenotype. Plastid Degs play roles in protecting the vital photosynthetic machinery and ptPDCs provide acetyl-CoA and NADH for fatty acid biosynthesis in plastids, which were different from functions of other isolated maize leaf color associated genes. The other two genes in the deletion were possibly associated with DNA repair and disease resistance, respectively. The pgl-sd mutation decreased contents of chlorophyll a, chlorophyll b, carotenoids by 37.2%, 22.1%, and 59.8%, respectively, and led to abnormal chloroplast. RNA-seq revealed that the transcription of several other genes involved in the structure and function of chloroplast was affected in the mutant. Conclusions It was identified that a 137.8-kb deletion causes the pgl-sd phenotype. Three genes in this deletion were possibly related to the chloroplast development, which may play roles different from that of other isolated maize leaf color associated genes.

Cadmium (Cd) is a toxic heavy metal that accumulates in plants, negatively affecting their physiological processes, growth, and development, and poses a threat to human health through the food chain. 6-phosphogluconolactonase (PGL) is a key enzyme in the Oxidative Pentose Phosphate Pathway(OPPP) in plant cells, essential for cellular metabolism. The OPPP pathway provides energy and raw materials for organisms and is involved in antioxidant reactions, lipid metabolism, and DNA synthesis. This study describes the Cd responsive gene AetPGL from Aegilops tauschii. Overexpression of AetPGL under Cd stress increased main root length and germination rate in Arabidopsis. Transgenic lines showed higher antioxidant enzyme activities and lower malondialdehyde (MDA) content compared to the wild type. The transgenic Arabidopsis accumulated more Cd in the aboveground part but not in the underground part. Expression levels of AtHMA3, AtNRAMP5, and AtZIP1 in the roots of transgenic plants increased under Cd stress, suggesting AetPGL may enhance Cd transport from root to shoot. Transcriptome analysis revealed enrichment of differentially expressed genes (DEGs) in the plant hormone signal transduction pathway in AetPGL-overexpressing plants. Brassinosteroids (BR), Gibbenellin acid (GA), and Jasmonic acid (JA) contents significantly increased after Cd treatment. These results indicate that AetPGL may enhance Arabidopsis’ tolerance to Cd by modulating plant hormone content. In conclusion, AetPGL plays a critical role in improving cadmium tolerance and accumulation and mitigating oxidative stress by regulating plant hormones, providing insights into the molecular mechanisms of plant Cd tolerance.

Vibrio vulnificus is an opportunistic, global pathogen that naturally inhabits sea water and is responsible for most vibriosis-related deaths. We investigated the genetic characteristics of V. vulnificus isolated from the clinical blood culture specimen of a patient with hepatitis B virus cirrhosis in 2018 (named as V. vulnificus VV2018) by whole genome sequencing (WGS). VV2018 belonged to a novel sequencing type 620 (ST620) and comprised two circular chromosomes, containing 4,389 potential coding sequences (CDSs) and 152 RNA genes. The phylogenetic tree of single nucleotide polymorphisms (SNPs) using 26 representative genomes revealed that VV2108 grouped with two other V. vulnificus strains isolated from humans. The pan-genome of V. vulnificus was constructed using 26 representative genomes to elucidate their genetic diversity, evolutionary characteristics, and virulence and antibiotic resistance profiles. The pan-genome analysis revealed that VV2018 shared a total of 3,016 core genes (≥99% presence), including 115 core virulence factors (VFs) and 5 core antibiotic resistance-related genes, and 309 soft core genes (≥95 and <99% presence) with 25 other V. vulnificus strains. The varG gene might account for the cefazolin resistance, and comparative analysis of the genetic context of varG revealed that two genes upstream and downstream of varG were conserved. The glycosylation (pgl) like genes were found in VV2018 compared with Pgl-related proteins in Neisseria that might affect the adherence of the strain in hosts. The comparative analysis of VV2018 would contribute to a better understanding of the virulence and antibiotic resistance profiles of V. vulnificus. Meanwhile much work remains to be done to better understand the function of pgl-like genes in V. vulnificus. [ABSTRACT FROM AUTHOR]

Background: The structural basis of chloroplast and the regulation of chloroplast biogenesis remain largely unknown in maize. Gene mutations in these pathways have been linked to the abnormal leaf color phenotype observed in some mutants. Large scale structure variants (SVs) are crucial for genome evolution, but few validated SVs have been reported in maize and little is known about their functions though they are abundant in maize genomes. Results: In this research, a spontaneous maize mutant, pale green leaf-shandong (pgl-sd), was studied. Genetic analysis showed that the phenotype of pale green leaf was controlled by a recessive Mendel factor mapped to a 156.8-kb interval on the chromosome 1 delineated by molecular markers gy546 and gy548. There were 7 annotated genes in this interval. Reverse transcription quantitative PCR analysis, SV prediction, and de novo assembly of pgl-sd genome revealed that a 137.8-kb deletion, which was verified by Sanger sequencing, might cause the pgl-sd phenotype. This deletion contained 5 annotated genes, three of which, including Zm00001eb031870, Zm00001eb031890 and Zm00001eb031900, were possibly related to the chloroplast development. Zm00001eb031870, encoding a Degradation of Periplasmic Proteins (Deg) homolog, and Zm00001eb031900, putatively encoding a plastid pyruvate dehydrogenase complex E1 component subunit beta (ptPDC-E1-β), might be the major causative genes for the pgl-sd mutant phenotype. Plastid Degs play roles in protecting the vital photosynthetic machinery and ptPDCs provide acetyl-CoA and NADH for fatty acid biosynthesis in plastids, which were different from functions of other isolated maize leaf color associated genes. The other two genes in the deletion were possibly associated with DNA repair and disease resistance, respectively. The pgl-sd mutation decreased contents of chlorophyll a, chlorophyll b, carotenoids by 37.2%, 22.1%, and 59.8%, respectively, and led to abnormal chloroplast. RNA-seq revealed that the transcription of several other genes involved in the structure and function of chloroplast was affected in the mutant. Conclusions: It was identified that a 137.8-kb deletion causes the pgl-sd phenotype. Three genes in this deletion were possibly related to the chloroplast development, which may play roles different from that of other isolated maize leaf color associated genes. [ABSTRACT FROM AUTHOR]

Purpose: Paraganglioma and pheochromocytoma are rare neuroendocrine tumors with severe metabolic and cardiovascular complications. Bladder PGLs are rare, and their clinical management is not precise. Here, we discuss the basic characteristics and perioperative management of bladder PGLs. Methods: We retrospectively reviewed 20 bladder PGL cases diagnosed at Sun Yat-sen University Cancer Center. Case notes were reviewed, clinical presentations, therapies, and outcomes were collected, and data analysis was performed. Results: Ten male and ten female patients with a median age of 47.5 years (range 14–69 years) were included. Most patients (65%) had no symptoms, and PGL was detected incidentally during medical checkups. All patients were treated surgically; 4 (20%) underwent transurethral resection of bladder tumor (TURBT), and 16 (80%) underwent partial cystectomy. Strong intraoperative blood pressure fluctuations were observed in 13 patients (65%). Two patients who were treated preoperatively with α-receptor blockers also experienced severe intraoperative blood pressure fluctuations. Postoperative measurements of troponin I were available for 3 patients, and all were significantly elevated. All patients were diagnosed with bladder PGL on postoperative pathological examination. The median follow-up time was 51 months (range 2–147 months), and 2 patients were lost to follow-up at 1 and 3 months; 16 (88.9%) survived without recurrence, 2 patients (11.1%) experienced recurrence, and 1 patient died. Conclusion: Most bladder paragangliomas are easily mistaken for bladder urothelial carcinoma, and robust hemodynamic instability during surgery might be a challenge for urologists. Postoperative monitoring of troponin I, regardless of the presence of clinical symptoms, is recommended for patients with bladder PGL. [ABSTRACT FROM AUTHOR]

Mycobacterium bovis bacillus Calmette-Guérin (BCG) efficacy as an immunotherapy tool can be influenced by the genetic background or immune status of the treated population and by the BCG substrain used. BCG comprises several substrains with genetic differences that elicit diverse phenotypic characteristics. Moreover, modifications of phenotypic characteristics can be influenced by culture conditions. However, several culture media formulations are used worldwide to produce BCG. To elucidate the influence of growth conditions on BCG characteristics, five different substrains were grown on two culture media, and the lipidic profile and physico-chemical properties were evaluated. Our results show that each BCG substrain displays a variety of lipidic profiles on the outermost surface depending on the growth conditions. These modifications lead to a breadth of hydrophobicity patterns and a different ability to reduce neutral red dye within the same BCG substrain, suggesting the influence of BCG growth conditions on the interaction between BCG cells and host cells.

Context. Pheochromocytomas and paragangliomas (PCC/PGLs) are diagnosed variously with increasing incidence and changing clinical and pathology pattern. Objective. The aim was to further characterize PCC/PGLs in a stable population. Methods. A retrospective, single institution study analysed adrenalectomies for PCC/PGLs between January 2010 - January 2019. Demographics, symptoms, blood pressure, preoperative hormones, imaging, histology, hospital stay, complications and three subgroups [based on the modality of diagnosis - incidentaloma group (IG), genetic group (GG) and symptomatic group (SG)] were noted. Results. 86 patients included IG 51 (59.3%), GG 10 (11.62%) and SG 25 patients (29.06%). Incidence was 5.30 cases/1 million population. 33.34% of the IG had a delayed diagnosis with a mean interval of 22.95 months (4- 120 months). Females presented more often with paroxysmal symptoms (PS) (p=0.011). Patients with PS and classic symptoms were younger (p=0.0087, p=0.0004) and those with PS required more inotropes postoperatively (p=0.014). SG had higher preoperative hormone levels (p=0.0048), larger tumors (p=0.0169) and more likely females. GG are younger compared with those from the IG (p=0.0001) or SG (p= 0.178). Conclusion. Majority of patients had an incidental and delayed diagnosis. If symptomatic, patients are more likely to be young females with higher hormone levels and larger tumors. [ABSTRACT FROM AUTHOR]

We study the involution of the real line, induced by Dyer's outer automorphism of PGL(2,Z). It is continuous at irrationals with jump discontinuities at rationals. We prove that its derivative exists almost everywhere and vanishes almost everywhere. [ABSTRACT FROM AUTHOR]

Abstract With the high number of patients infected by tuberculosis and the sharp increase of drug-resistant tuberculosis cases, developing new drugs to fight this disease has become increasingly urgent. In this context, analogs of the naturally occurring enolphosphates Cyclipostins and Cyclophostin (CyC analogs) offer new therapeutic opportunities. The CyC analogs display potent activity both in vitro and in infected macrophages against several pathogenic mycobacteria including Mycobacterium tuberculosis and Mycobacterium abscessus. Interestingly, these CyC inhibitors target several enzymes with active-site serine or cysteine residues that play key roles in mycobacterial lipid and cell wall metabolism. Among them, TesA, a putative thioesterase involved in the synthesis of phthiocerol dimycocerosates (PDIMs) and phenolic glycolipids (PGLs), has been identified. These two lipids (PDIM and PGL) are non-covalently bound to the outer cell wall in several human pathogenic mycobacteria and are important virulence factors. Herein, we used biochemical and structural approaches to validate TesA as an effective pharmacological target of the CyC analogs. We confirmed both thioesterase and esterase activities of TesA, and showed that the most active inhibitor CyC 17 binds covalently to the catalytic Ser104 residue leading to a total loss of enzyme activity. These data were supported by the X-ray structure, obtained at a 2.6 - Å resolution, of a complex in which CyC 17 is bound to TesA. Our study provides evidence that CyC 17 inhibits the activity of TesA, thus paving the way to a new strategy for impairing the PDIM and PGL biosynthesis, potentially decreasing the virulence of associated mycobacterial species. Graphical abstract Unlabelled Image Highlights • First 3D structure of M. tuberculosis thioesterase TesA involved in PDIM/PGL synthesis • First biochemical characterization of TesA • TesA possesses a lid protecting its active site in closed conformation. • CyC 17 , a Cyclipostins analog, is a potent and covalent inhibitor of TesA • Proteomic and structural studies revealed a chemical rearrangement on CyC 17 -core in TesA active site. [ABSTRACT FROM AUTHOR]

Objective To review the optimal techniques for localization and characterization of neck paragangliomas (PGL). Material and methods Systematic review of the literature from the PubMed/Medline database. Results Neck PGL are hypervascular tumours essentially arising from paraganglionic tissue situated at the carotid bifurcation (carotid body) and along the vagus nerve. Morphological and functional imaging are indicated to confirm the diagnosis, identify multifocal disease and for local and regional staging. MR angiography is the noninvasive technique of choice. CT scan and especially CT angiography are excellent alternatives for diagnosis and staging. Conventional arteriography remains useful preoperatively for embolization and occlusion tests. Functional imaging allows localization and characterization of PGLs. Somatostatin receptor scintigraphy (SRS) was the reference imaging technique for staging of sporadic PGLs. The indications for PET imaging have been extended over recent years in parallel with the development of new tracers such as [ 18 F]-FDOPA PET or 68 Gallium-labelled DOTA peptides. 68 Gallium-labelled DOTA peptides has become the first-line imaging modality in the evaluation of cervical PGLs, regardless of the genetic background. Conclusion Morphological and functional imaging is essential for the staging of neck PGL. [ABSTRACT FROM AUTHOR]

Studies have reported a wide range of inflammatory responses in the nerve, skin and plasma of leprosy patients. The expression levels of each biomolecule was individualistic, however could be categorized as high and low based on their statistical mean level. Here we report for the first time, expression of a set of biomolecules relating with each other in a defined proportion. The hypothesis of this paper is that the segregation of high and low combinations of a set of biomolecules follows either classical Mendelian dihybrid ratio or epistatic ratios. This hypothesis was tested for 17 molecules in three tissues; nerve, skin and plasma and were confirmed to interact in 9:7, 9:3:4, 12:3:1, 13:3, 15:1 epistatic proportions. These findings suggest that there could be a significant role of networking of molecules in defined epistatic proportions and could be important in pathophysiology of peripheral nerve. [ABSTRACT FROM AUTHOR]

To determine whether intact pks15/1 is unique to the W-Beijing family, we investigated 147 Mycobacterium tuberculosis strains with different IS6110 genotypes. Intact pks15/1 was found in 87.8% of cerebrospinal fluid and 84.9% of sputum isolates. It was found not only in W-Beijing strains (≈97%) but also in other genotypes (38.5%–100%).

The perpetual arms race between bacteria and phage has resulted in the evolution of efficient resistance systems that protect bacteria from phage infection. Such systems, which include the CRISPR-Cas and restriction-modification systems, have proven to be invaluable in the biotechnology and dairy industries. Here, we report on a six-gene cassette in Bacillus cereus which, when integrated into the Bacillus subtilis genome, confers resistance to a broad range of phages, including both virulent and temperate ones. This cassette includes a putative Lon-like protease, an alkaline phosphatase domain protein, a putative RNA-binding protein, a DNA methylase, an ATPase-domain protein, and a protein of unknown function. We denote this novel defense system BREX (Bacteriophage Exclusion) and show that it allows phage adsorption but blocks phage DNA replication. Furthermore, our results suggest that methylation on non-palindromic TAGGAG motifs in the bacterial genome guides self/non-self discrimination and is essential for the defensive function of the BREX system. However, unlike restriction-modification systems, phage DNA does not appear to be cleaved or degraded by BREX, suggesting a novel mechanism of defense. Pan genomic analysis revealed that BREX and BREX-like systems, including the distantly related Pgl system described in Streptomyces coelicolor, are widely distributed in ~10% of all sequenced microbial genomes and can be divided into six coherent subtypes in which the gene composition and order is conserved. Finally, we detected a phage family that evades the BREX defense, implying that anti- BREX mechanisms may have evolved in some phages as part of their arms race with bacteria. [ABSTRACT FROM AUTHOR]

Positron emission tomography (PET) is used for staging and response evaluation in primary gastric lymphoma (PGL). However, the implications of [F]-2-fluoro-2-deoxy- d-glucose (F-FDG) uptake in PGL at first diagnosis have not been reported. The relationship between F-FDG uptake and the expression of facilitative glucose transporters (GLUTs), hexokinase II (HK II), and Ki67, as well as malignant potential in PGL, was assessed in this study. We analyzed 23 patients with PGL [nine with diffuse large B-cell lymphoma (DLBCL); seven with high-grade mucosa-associated lymphoid tissue (MALT) lymphoma; and seven with low-grade MALT lymphoma]. The expression levels of GLUT1, GLUT3, HK II, and Ki67 were evaluated according to the percentage of positive area determined by immunohistochemistry. Standardized uptake values correlated significantly with pathological malignant potentials (low-grade/high-grade MALT lymphoma and DLBCL: p = 0.001-0.002), Ki67 ( p < 0.001), and GLUT1 expression ( p = 0.02). We determined that F-FDG uptake is related to GLUT1 expression and tumor histological grade as well as Ki67 in PGL. [ABSTRACT FROM AUTHOR]

Abstract: Mycobacterium bovis is an important pathogen of both domesticated and wild animals in many countries, and improved vaccines have great potential to assist in its control and eventual eradication. One of the hallmarks of members of the Mycobacterium tuberculosis complex, which includes both M. bovis and M. tuberculosis, is their ability to synthesise an impressive array of unique and complex lipids, many of which act as defensive, offensive or adaptive effectors of virulence. For example, studies focussed on the development of rationally attenuated strains of both M. bovis and M. tuberculosis with efficacy as animal or human vaccines have shown that the phthiocerol dimycocerosates (PDIMs) and glycosylphenol-PDIM (phenolic glycolipid, PGL) are key virulence factors. The availability of the genome sequences for M. bovis and M. tuberculosis, together with mutants of these organisms carrying defects in lipid biosynthesis, and biochemical and molecular tools to dissect lipid biosynthesis pathways, has enabled developments in our understanding of the biosynthesis of PDIMs and PGL, as well as the possible roles played by PDIMs and PGL in virulence. In this article we review some of these developments, and also propose a cryptic lipid biosynthesis pathway in M. bovis and M. tuberculosis that may be involved in the production of an unrecognised, virulence-associated lipopeptide. [Copyright &y& Elsevier]

Protein glycosylation is an important element of biologic systems because of its significant effects on protein properties and functions. Although prominent within all domains of life, 0-linked glycosylation systems modifying serine and threonine residues within bacteria and eukaryotes differ substantially in target protein selectivity. In particular, well-characterized bacterial systems have been invariably dedicated to modification of individual proteins or related subsets thereof. Here we characterize a general 0-linked glycosylation system that targets structurally and functionally diverse groups of membrane-associated proteins in the Gram-negative bacterium Neisseria gonorrhoeae. the etiologic agent of the human disease gonorrhea. The 11 glycoproteins identified here are implicated in activities as varied as protein folding, disulfide bond formation, and solute uptake, as well as both aerobic and anaerobic respiration. Along with their common trafficking within the periplasmic compartment, the protein substrates share quasi-related domains bearing signatures of low complexity that were demonstrated to encompass sites of glycan occupancy. Thus, as in eukaryotes, the broad scope of this system is dictated by the relaxed specificity of the glycan transferase as well as the bulk properties and context of the proteintargeting signal rather than by a strict amino acid consensus sequence. Together, these findings reveal previously unrecognized commonalities linking 0-linked protein glycosylation in distantly related life forms. [ABSTRACT FROM AUTHOR]

Objective: Carotid body tumour (CBT) is a rare but the most common form of head and neck paraganglioma (PGL). We present the biggest ever series on CBT in UK/EU discussing diagnostic challenges, surgical treatment and complications of surgical intervention. Method: A detailed proforma was designed and sent to all members of Joint Vascular Reasearch Group (JVRG). Data of 95 patients was collected. Generic terms including carotid body tumour/s, or paraganglioma/s were used to search a variety of electronic database in order to get latest informations available in literature. Results: A total of 95 patients were recorded in our data from 1979 to 2005. Mean age of presentation was 55 years. Incidence was higher in females. CBT was more common on right side (58%). 18% tumours were bilateral. Neck lump (98%) and pressure symptoms including cranial nerve deficits and pain were main presenting complaints. About 18% of tumours were familial. Only 4.2% were malignant. Duplex scan is the best investigation for diagnosis, though MRI, DSA and CT scan are important for preoperative assessment. Surgery is the treatment of choice. Stroke and cranial nerve injury constitute postoperative morbidity (35%) and mortality (1%). Incidence of postoperative cranial nerve deficit was about 19%. Combined ipsilateral and contralateral recurrence rate was 4.2%. Conclusion: CBT is a rare condition which needs surgical excision by experienced vascular surgeon. Surgical resection is associated with significant morbidity of 35% and mortality of 1%. Mostly CBT is benign but malignant forms are not uncommon. [Copyright &y& Elsevier]

Although deceptively simple, the etio-pathogenesis of pheochromocytoma represents a clinical and molecular genetic investigative challenge. Here, we summarize, from a historical point of view, the 22-year-long studies initiated at the University of Freiburg, which developed from a local experience to a national and finally an international effort. All research activities are translational and clinical and hence, registry based and intended to improve the outcome of the patients, whether by improved detection, prevention, or treatment. Major clinical steps are the prospective study on hormone tests and imaging techniques for adrenal and extra-adrenal abdominal tumors as well as the concept of organ sparing and endoscopic tumor resection. Further, we introduced 18-fluoro-dopa positron emission tomography. Population-based registries were used in order to identify germline mutations in the susceptibility genes VHL, RET, SDHB, and SDHD in non-syndromic pheochromocytoma. We differentiated distinct clinical features of paraganglioma syndromes associated with SDHB and SDHD gene mutations. Finally, we identified predictors and prevalence of paraganglioma syndromes associated with mutations of the SDHC gene. [ABSTRACT FROM AUTHOR]

Abstract: The pentose-phosphate pathway (PPP) is an important part of central metabolism in many organisms. A pgl− mutation that decreases the efficiency of the second stage of PPP has been described and mapped at approx. 17.2min of the Escherichia coli chromosome more than 30 years ago. Although it has recently been shown that deletion of ORF ybhE leads to earlier detected Pgl− phenotype for E. coli mutant strain, 6-phosphogluconolactonase from this organism has not been characterized. In the present, independent investigation we show that the Pgl− phenotype of ΔybhE MG1655 could be complemented by insertion of the well-characterized pgl gene from Pseudomonas putida whose protein product has no visible homology with E. coli YbhE. Moreover, a final confirmation that ybhE actually encodes 6PGL in E. coli was obtained through overexpression of the cloned gene, purification of the protein product, followed by direct determination of its enzymatic activity in vitro. [Copyright &y& Elsevier]

An extracellular endo-polygalacturonase (PGL) from the psychrophilic fungus, Mucor flavus was purified to homogeneity by two ion-exchange chromatography steps using CM-Sepharose CL 6B. The PGL with a molecular mass of 40 kDa by SDS–PAGE, 38.7 kDa by mass spectrometry and a pI above 8.3 was optimally active at 45 °C and between pH 3.5 and 5.5. It was stable up to 40 °C for 4 h and between pH 2.5 and 6.0 for 20 h at 20 °C. Activity and stability of PGL decreased rapidly above 50 °C. The PGL was active on polygalacturonic acid and esterified pectin, but the activity on pectin decreased with an increase in degree of esterification. The enzyme rapidly decreased the viscosity of polygalacturonic acid and 89% esterified pectin, but the amount of reducing sugars released from polygalacturonic acid was double that of from 89% esterified pectin. The PGL released a series of oligo-galacturonates from polygalacturonic acid and pectin. The concentration of oligo-galacturonates with DP above 5 was initially high and decreased with time. The enzyme preferentially hydrolysed the oligo-galacturonates with DP 5 and above in an endo-fashion and appeared to possess 6–7 sugar binding sites in its active site. A 39 amino acid sequence at the N-terminal of this PGL showed only 17–29% sequence identity with PGLs of glycoside hydrolase family 28. [Copyright &y& Elsevier]

Mycobacterium bovis bacillus Calmette-Guérin (BCG) efficacy as an immunotherapy tool can be influenced by the genetic background or immune status of the treated population and by the BCG substrain used. BCG comprises several substrains with genetic differences that elicit diverse phenotypic characteristics. Moreover, modifications of phenotypic characteristics can be influenced by culture conditions. However, several culture media formulations are used worldwide to produce BCG. To elucidate the influence of growth conditions on BCG characteristics, five different substrains were grown on two culture media, and the lipidic profile and physico-chemical properties were evaluated. Our results show that each BCG substrain displays a variety of lipidic profiles on the outermost surface depending on the growth conditions. These modifications lead to a breadth of hydrophobicity patterns and a different ability to reduce neutral red dye within the same BCG substrain, suggesting the influence of BCG growth conditions on the interaction between BCG cells and host cells. [ABSTRACT FROM AUTHOR]

Summary: Paragangliomas are rare tumours of the autonomic nervous system that occur in both sporadic and hereditary forms. They are usually benign tumours with low mortality, but can cause significant morbidity related to mass effect. Genetic predisposition to develop paraganglioma can occur within known tumour syndromes and familial tumours tend to present at a younger age and at multiple sites compared to sporadic tumours. Tumours should be diagnosed and excised as early as possible, as studies have shown morbidity to be directly related to tumour size. We present a case of a 14-year-old boy with multiple paraganglioma and a strong family history of paraganglioma. He suffered significant morbidity at resection of an extra-adrenal retroperitoneal tumour due to late diagnosis and was later unable to undergo excision of a head and neck paraganglioma due to its size and relation to neurovascular structures in the neck. We review the current literature on suggested genetic counselling (psychological counselling and DNA analysis) and radiological screening guidelines and recommend that genetic counselling should be offered to all patients with a family history of paraganglioma from the age of 5 years. Those positive for paternal paraganglioma locus gene should then undergo regular radiological screening with MRI. [Copyright &y& Elsevier]

The genus Neisseria includes three major species of importance to human health and disease ( Neisseria gonorrhoeae , Neisseria meningitidis , and Neisseria lactamica ) that express broad-spectrum O -linked protein glycosylation (Pgl) systems. The potential for related Pgl systems in other species in the genus, however, remains to be determined. Using a strain of Neisseria elongata subsp. glycolytica , a unique tetrasaccharide glycoform consisting of di- N -acetylbacillosamine and glucose as the first two sugars followed by a rare sugar whose mass spectrometric fragmentation profile was most consistent with di- N -acetyl hexuronic acid and a N -acetylhexosamine at the nonreducing end has been identified. Based on established mechanisms for UDP-di- N -acetyl hexuronic acid biosynthesis found in other microbes, we searched for genes encoding related pathway components in the N. elongata subsp. glycolytica genome. Here, we detail the identification of such genes and the ensuing glycosylation phenotypes engendered by their inactivation. While the findings extend the conservative nature of microbial UDP-di- N -acetyl hexuronic acid biosynthesis, mutant glycosylation phenotypes reveal unique, relaxed specificities of the glycosyltransferases and oligosaccharyltransferases to incorporate pathway intermediate UDP-sugars into mature glycoforms. IMPORTANCE Broad-spectrum protein glycosylation (Pgl) systems are well recognized in bacteria and archaea. Knowledge of how these systems relate structurally, biochemically, and evolutionarily to one another and to others associated with microbial surface glycoconjugate expression is still incomplete. Here, we detail reverse genetic efforts toward characterization of protein glycosylation mutants of N. elongata subsp. glycolytica that define the biosynthesis of a conserved but relatively rare UDP-sugar precursor. The results show both a significant degree of intra- and transkingdom conservation in the utilization of UDP-di- N -acetyl-glucuronic acid and singular properties related to the relaxed specificities of the N. elongata subsp. glycolytica system., (Copyright © 2018 American Society for Microbiology.)

Objective: To report the design and implementation of the Right Drug, Right Dose, Right Time-Using Genomic Data to Individualize Treatment protocol that was developed to test the concept that prescribers can deliver genome-guided therapy at the point of care by using preemptive pharmacogenomics (PGx) data and clinical decision support (CDS) integrated into the electronic medical record (EMR)., Patients and Methods: We used a multivariate prediction model to identify patients with a high risk of initiating statin therapy within 3 years. The model was used to target a study cohort most likely to benefit from preemptive PGx testing among the Mayo Clinic Biobank participants, with a recruitment goal of 1000 patients. We used a Cox proportional hazards model with variables selected through the Lasso shrinkage method. An operational CDS model was adapted to implement PGx rules within the EMR., Results: The prediction model included age, sex, race, and 6 chronic diseases categorized by the Clinical Classifications Software for International Classification of Diseases, Ninth Revision codes (dyslipidemia, diabetes, peripheral atherosclerosis, disease of the blood-forming organs, coronary atherosclerosis and other heart diseases, and hypertension). Of the 2000 Biobank participants invited, 1013 (51%) provided blood samples, 256 (13%) declined participation, 555 (28%) did not respond, and 176 (9%) consented but did not provide a blood sample within the recruitment window (October 4, 2012, through March 20, 2013). Preemptive PGx testing included CYP2D6 genotyping and targeted sequencing of 84 PGx genes. Synchronous real-time CDS was integrated into the EMR and flagged potential patient-specific drug-gene interactions and provided therapeutic guidance., Conclusion: This translational project provides an opportunity to begin to evaluate the impact of preemptive sequencing and EMR-driven genome-guided therapy. These interventions will improve understanding and implementation of genomic data in clinical practice., (Copyright © 2014 Mayo Foundation for Medical Education and Research. Published by Elsevier Inc. All rights reserved.)

This study aims to investigate the feasibility of Levodopa transdermal delivery systems (TDSs). Levodopa TDSs were formulated using various vehicles and permeation enhancers, and in vitro permeation and in vivo pharmacokinetic studies were carried out. In the in vitro study, ester-type vehicles showed relatively high enhancing effects; propylene glycol monocaprylate and propylene glycol monolaurate showed the highest permeation fluxes from both solution and pressure sensitive adhesive (PSA) TDS formulations. Lag time was dramatically shortened with PSA TDS formulations as compared with solution formulations. In the in vivo study, the addition of fatty acids increased blood drug concentrations regardless of the kind or concentration of fatty acid; the AUCinf increased up to 8.7 times as compared with propylene glycol (PG) alone. PSA TDS containing 10% linoleic acid exhibited prolonged Tmax as compared with oral form. Total clearance of L-dopa from PSA TDSs was significantly lower than from oral form (up to 86.8 times). Especially, PSA TDS containing 10% linoleic acid (LOA) revealed 76.2 fold higher AUCinf than oral administration. Based on our results, the L-dopa PSA TDS containing PG with 10% LOA could be used as a good adjuvant therapy for Parkinson's disease patients who experience symptom fluctuation by L-dopa oral administration., (Copyright © 2013 Elsevier B.V. All rights reserved.)