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1.
PLoS Genet ; 18(3): e1010114, 2022 03.
Artigo em Inglês | MEDLINE | ID: mdl-35298461

RESUMO

The highly evolutionarily conserved transport protein particle (TRAPP) complexes (TRAPP II and III) perform fundamental roles in subcellular trafficking pathways. Here we identified biallelic variants in TRAPPC10, a component of the TRAPP II complex, in individuals with a severe microcephalic neurodevelopmental disorder. Molecular studies revealed a weakened interaction between mutant TRAPPC10 and its putative adaptor protein TRAPPC2L. Studies of patient lymphoblastoid cells revealed an absence of TRAPPC10 alongside a concomitant absence of TRAPPC9, another key TRAPP II complex component associated with a clinically overlapping neurodevelopmental disorder. The TRAPPC9/10 reduction phenotype was recapitulated in TRAPPC10-/- knockout cells, which also displayed a membrane trafficking defect. Notably, both the reduction in TRAPPC9 levels and the trafficking defect in these cells could be rescued by wild type but not mutant TRAPPC10 gene constructs. Moreover, studies of Trappc10-/- knockout mice revealed neuroanatomical brain defects and microcephaly, paralleling findings seen in the human condition as well as in a Trappc9-/- mouse model. Together these studies confirm autosomal recessive TRAPPC10 variants as a cause of human disease and define TRAPP-mediated pathomolecular outcomes of importance to TRAPPC9 and TRAPPC10 mediated neurodevelopmental disorders in humans and mice.


Assuntos
Microcefalia , Transtornos do Neurodesenvolvimento , Animais , Humanos , Camundongos , Microcefalia/genética , Transtornos do Neurodesenvolvimento/genética , Fenótipo
2.
J Gene Med ; 25(10): e3513, 2023 10.
Artigo em Inglês | MEDLINE | ID: mdl-37178061

RESUMO

BACKGROUND: Population diversity is important and rare disease isolates can frequently reveal novel homozygous or biallelic mutations that lead to expanded clinical heterogeneity, with diverse clinical presentations. METHODS: The present study describes two consanguineous families with a total of seven affected individuals suffering from a clinically similar severe syndromic neurological disorder, with abnormal development and central nervous system (CNS) and peripheral nervous system (PNS) abnormalities. Whole exome sequencing (WES) and Sanger sequencing followed by 3D protein modeling was performed to identify the disease-causing gene. RNA was extracted from the fresh blood of both families affected and healthy individuals. RESULTS: The families were clinically assessed in the field in different regions of Khyber Pakhtunkhwa. Magnetic resonance imagining was obtained in the probands and blood was collected for DNA extraction and WES was performed. Sanger sequencing confirmed a homozygous, likely pathogenic mutation (GRCh38: chr17:42684199G>C; (NM_003632.3): c.333G>C);(NP_003623.1): p.(Trp111Cys) in the CNTNAP1 gene in family A, previously associated with Congenital Hypo myelinating Neuropathy 3 (CHN3; OMIM # 618186) and a novel nonsense variant in family B, (GRCh38: chr16: 57654086C>T; NC_000016.10 (NM_001370440.1): c.721C>T); (NP_001357369.1): p.(Gln241Ter) in the ADGRG1 gene previously associated with bilateral frontoparietal polymicrogyria (OMIM # 606854); both families have extended CNS and PNS clinical manifestations. In addition, 3D protein modeling was performed for the missense variant, p.(Trp111Cys), identified in the CNTNAP1, suggesting extensive secondary structure changes that might lead to improper function or downstream signaling. No RNA expression was observed in both families affected and healthy individuals hence showing that these genes are not expressed in blood. CONCLUSIONS: In the present study, two novel biallelic variants in the CNTNAP1 and ADGRG1 genes in two different consanguineous families with a clinical overlap in the phenotype were identified. Thus, the clinical and mutation spectrum is expanded to provide further evidence that CNTNAP1 and ADGRG1 are very important for widespread neurological development.


Assuntos
Moléculas de Adesão Celular Neuronais , Mutação de Sentido Incorreto , Humanos , Consanguinidade , Mutação , Genes Recessivos , Fenótipo , Moléculas de Adesão Celular Neuronais/genética
3.
Pak J Med Sci ; 39(4): 1124-1128, 2023.
Artigo em Inglês | MEDLINE | ID: mdl-37492301

RESUMO

Objective: To identify genetic causes of Shabbir syndrome in two patients of Pakistani origin. Methods: In the present study, we have investigated a Pakistani family with two affected members segregating Laryngo-onycho-cutaneous (LOC) syndrome. The patients were diagnosed as suspected cases of LOC based on phenotypes including abnormal larynx, nails, and hyperpigmentation in patients' eyes. Genetic investigation was done by performing whole exome sequencing (WES) using DNA of the patients. Sanger sequencing was performed to validate WES findings and segregation analysis in the family. Results: Data analysis of exomes and Sanger sequencing of patients revealed a homozygous one base pair duplication (NM_000227.6; LAMA3; c.151dup; p.Val51GlyfsTer4) in LAMA3 in the patients. Parents of the patients were heterozygous for the identified variant. Conclusion: Previously, the same variant has been found in most of the Pakistani Punjabi patients affected with LOC. Therefore, Pakistani Punjabi families affected with Shabbir Syndrome may be screened for c.151dup variant in LAMA3 using targeted sequencing. Sanger sequencing is a cost-effective and time-saving technique as compared to whole exome/genome sequencing. Hence, developing ethnicity-specific LAMA3 targeted molecular diagnostic test would be cost-effective. Further, the study would assist in carrier testing and prenatal diagnosis of the affected families.

4.
J Gene Med ; 24(4): e3412, 2022 04.
Artigo em Inglês | MEDLINE | ID: mdl-35075722

RESUMO

BACKGROUND: Limb-girdle muscular dystrophy (LGMD) comprises a heterogeneous group of diseases, affecting different muscles, predominantly skeletal muscles and cardiac muscles of the body. LGMD is classified into two main subtypes A and B, which are further subclassified into eight dominant and thirty recessive subtypes. Three genes, namely POPDC1, POPDC2 and POPDC3, encode popeye domain-containing protein (POPDC), and the variants of POPDC1 and POPDC3 genes have been associated with LGMD. METHODS: In the present study, we performed whole-exome sequencing (WES) analysis on a single-family to investigate the hallmark features of LGMD. The results of WES were further confirmed by Sanger sequencing and 3D protein modeling was also conducted. RESULTS: WES data analysis and Sanger sequencing revealed a homozygous missense variant (c.460A>G; p.Lys154Glu) at a highly conserved amino acid position in the POPDC3. Mutations in the POPDC3 gene have been previously associated with recessive limb-girdle muscular dystrophy type 26. 3D protein modeling further suggested that the identified variant might affect the POPDC3 structure and proper function. CONCLUSIONS: The present study confirms the role of POPDC3 in LGMD, and will facilitate genetic counseling of the family to mitigate the risks of the carrier or affects on future pregnancies.


Assuntos
Moléculas de Adesão Celular , Proteínas Musculares , Distrofia Muscular do Cíngulo dos Membros , Moléculas de Adesão Celular/genética , Homozigoto , Humanos , Proteínas Musculares/genética , Músculo Esquelético , Distrofia Muscular do Cíngulo dos Membros/genética , Mutação , Mutação de Sentido Incorreto
5.
Mol Biol Rep ; 47(10): 7655-7673, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32979167

RESUMO

The salinity stress tolerance in plants has been studied enormously, reflecting its agronomic relevance. Despite the extensive research, limited success has been achieved in relation to the plant tolerance mechanism. The beneficial interaction between Piriformospora indica and rice could essentially improve the performance of the plant during salt stress. In this study, the transcriptomic data between P. indica treated and untreated rice roots were compared under control and salt stress conditions. Overall, 661 salt-responsive differentially expressed genes (DEGs) were detected with 161 up- and 500 down-regulated genes in all comparison groups. Gene ontology analyses indicated the DEGs were mainly enriched in "auxin-activated signaling pathway", "water channel activity", "integral component of plasma membrane", "stress responses", and "metabolic processes". Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed that the DEGs were primarily related to "Zeatin biosynthesis", "Fatty acid elongation", "Carotenoid biosynthesis", and "Biosynthesis of secondary metabolites". Particularly, genes related to cell wall modifying enzymes (e.g. invertase/pectin methylesterase inhibitor protein and arabinogalactans), phytohormones (e.g. Auxin-responsive Aux/IAA gene family, ent-kaurene synthase, and 12-oxophytodienoate reductase) and receptor-like kinases (e.g. AGC kinase and receptor protein kinase) were induced in P. indica colonized rice under salt stress condition. The differential expression of these genes implies that the coordination between hormonal crosstalk, signaling, and cell wall dynamics contributes to the higher growth and tolerance in P. indica-inoculated rice. Our results offer a valuable resource for future functional studies on salt-responsive genes that should improve the resilience and adaptation of rice against salt stress.


Assuntos
Basidiomycota/metabolismo , Endófitos/metabolismo , Perfilação da Expressão Gênica , Regulação Fúngica da Expressão Gênica , Oryza/microbiologia , Salinidade , Tolerância ao Sal
6.
Ann Hum Genet ; 83(6): 477-482, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31173343

RESUMO

Ciliopathies are a clinically and genetically heterogeneous group of disorders often exhibiting phenotypic overlap and caused by abnormalities in the structure or function of cellular cilia. As such, a precise molecular diagnosis is important for guiding clinical management and genetic counseling. In the present study, two Pakistani families comprising individuals with overlapping clinical features suggestive of a ciliopathy syndrome, including intellectual disability, obesity, congenital retinal dystrophy, and hypogonadism (in males), were investigated clinically and genetically. Whole-exome sequencing identified the likely causes of disease as a novel homozygous frameshift mutation (NM_152384.2: c.196delA; p.(Arg66Glufs*12); family 1) in BBS5, and a nonsense mutation (NM_019892.5:c.1879C>T; p.Gln627*; family 2) in INPP5E, previously reported in an extended Pakistani family with MORM syndrome. Our findings expand the molecular spectrum associated with BBS5 mutations in Pakistan and provide further supportive evidence that the INPP5E mutation is a common cause of ciliopathy in Northern Pakistan, likely representing a regional founder mutation. This study also highlights the value of genomic studies in Pakistan for families affected by rare heterogeneous developmental disorders and where clinical phenotyping may be limited by geographical and financial constraints. The identification of the spectrum and frequency of disease-causing variants within this setting enables the development of population-specific genetic testing strategies targeting variants common to the local population and improving health care outcomes.


Assuntos
Ciliopatias/diagnóstico , Ciliopatias/genética , Proteínas do Citoesqueleto/genética , Mutação , Proteínas de Ligação a Fosfato/genética , Monoéster Fosfórico Hidrolases/genética , Adolescente , Adulto , Criança , Pré-Escolar , Análise Mutacional de DNA , Feminino , Efeito Fundador , Estudos de Associação Genética/métodos , Predisposição Genética para Doença , Humanos , Lactente , Masculino , Pessoa de Meia-Idade , Paquistão , Linhagem , Adulto Jovem
7.
BMC Med Genet ; 20(1): 199, 2019 12 18.
Artigo em Inglês | MEDLINE | ID: mdl-31852446

RESUMO

BACKGROUND: Neurological disorders are a common cause of morbidity and mortality within Pakistani populations. It is one of the most important challenges in healthcare, with significant life-long socio-economic burden. METHODS: We investigated the cause of disease in three Pakistani families in individuals with unexplained autosomal recessive neurological conditions, using both genome-wide SNP mapping and whole exome sequencing (WES) of affected individuals. RESULTS: We identified a homozygous splice site variant (NM_000521:c.445 + 1G > T) in the hexosaminidase B (HEXB) gene confirming a diagnosis of Sandhoff disease (SD; type II GM2-gangliosidosis), an autosomal recessive lysosomal storage disorder caused by deficiency of hexosaminidases in a single family. In two further unrelated families, we identified a homozygous frameshift variant (NM_024298.3:c.758_778del; p.Glu253_Ala259del) in membrane-bound O-acyltransferase family member 7 (MBOAT7) as the likely cause of disease. MBOAT7 gene variants have recently been identified as a cause of intellectual disability (ID), seizures and autistic features. CONCLUSIONS: We identified two metabolic disorders of lipid biosynthesis within three Pakistani families presenting with undiagnosed neurodevelopmental conditions. These findings enabled an accurate neurological disease diagnosis to be provided for these families, facilitating disease management and genetic counselling within this population. This study consolidates variation within MBOAT7 as a cause of neurodevelopmental disorder, broadens knowledge of the clinical outcomes associated with MBOAT7-related disorder, and confirms the likely presence of a regionally prevalent founder variant (c.758_778del; p.Glu253_Ala259del) in Pakistan.


Assuntos
Aciltransferases/genética , Homozigoto , Proteínas de Membrana/genética , Doenças do Sistema Nervoso/genética , Cadeia beta da beta-Hexosaminidase/genética , Consanguinidade , Eletroencefalografia , Feminino , Genes Recessivos , Humanos , Lactente , Imageamento por Ressonância Magnética , Masculino , Mutação , Doenças do Sistema Nervoso/diagnóstico por imagem , Doenças do Sistema Nervoso/fisiopatologia , Paquistão , Polimorfismo de Nucleotídeo Único , Sequenciamento do Exoma
8.
J Biol Chem ; 290(15): 9626-45, 2015 Apr 10.
Artigo em Inglês | MEDLINE | ID: mdl-25713147

RESUMO

The essential mycobacterial protein kinases PknA and PknB play crucial roles in modulating cell shape and division. However, the precise in vivo functional aspects of PknA have not been investigated. This study aims to dissect the role of PknA in mediating cell survival in vitro as well as in vivo. We observed aberrant cell shape and severe growth defects when PknA was depleted. Using the mouse infection model, we observe that PknA is essential for survival of the pathogen in the host. Complementation studies affirm the importance of the kinase, juxtamembrane, and transmembrane domains of PknA. Surprisingly, the extracytoplasmic domain is dispensable for cell growth and survival in vitro. We find that phosphorylation of the activation loop at Thr(172) of PknA is critical for bacterial growth. PknB has been previously suggested to be the receptor kinase, which activates multiple kinases, including PknA, by trans-phosphorylating their activation loop residues. Using phospho-specific PknA antibodies and conditional pknB mutant, we find that PknA autophosphorylates its activation loop independent of PknB. Fluorescently tagged PknA and PknB show distinctive distribution patterns within the cell, suggesting that although both kinases are known to modulate cell shape and division, their modes of action are likely to be different. This is supported by our findings that expression of kinase-dead PknA versus kinase-dead PknB in mycobacterial cells leads to different cellular phenotypes. Data indicate that although PknA and PknB are expressed as part of the same operon, they appear to be regulating cellular processes through divergent signaling pathways.


Assuntos
Proteínas de Bactérias/metabolismo , Viabilidade Microbiana , Mycobacterium tuberculosis/enzimologia , Proteínas Serina-Treonina Quinases/metabolismo , Animais , Proteínas de Bactérias/genética , Biocatálise , Western Blotting , Ativação Enzimática , Feminino , Interações Hospedeiro-Patógeno , Masculino , Camundongos Endogâmicos C57BL , Microscopia Eletrônica de Varredura , Microscopia de Fluorescência , Mutação , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/fisiologia , Fosforilação , Proteínas Serina-Treonina Quinases/genética , Tuberculose/microbiologia
9.
J Biol Chem ; 289(20): 13858-75, 2014 May 16.
Artigo em Inglês | MEDLINE | ID: mdl-24706757

RESUMO

The Mycobacterium tuberculosis protein kinase B (PknB) comprises an intracellular kinase domain, connected through a transmembrane domain to an extracellular region that contains four PASTA domains. The present study describes the comprehensive analysis of different domains of PknB in the context of viability in avirulent and virulent mycobacteria. We find stringent regulation of PknB expression necessary for cell survival, with depletion or overexpression of PknB leading to cell death. Although PknB-mediated kinase activity is essential for cell survival, active kinase lacking the transmembrane or extracellular domain fails to complement conditional mutants not expressing PknB. By creating chimeric kinases, we find that the intracellular kinase domain has unique functions in the virulent strain, which cannot be substituted by other kinases. Interestingly, we find that although the presence of the C-terminal PASTA domain is dispensable in the avirulent M. smegmatis, all four PASTA domains are essential in M. tuberculosis. The differential behavior of PknB vis-à-vis the number of essential PASTA domains and the specificity of kinase domain functions suggest that PknB-mediated growth and signaling events differ in virulent compared with avirulent mycobacteria. Mouse infection studies performed to determine the role of PknB in mediating pathogen survival in the host demonstrate that PknB is not only critical for growth of the pathogen in vitro but is also essential for the survival of the pathogen in the host.


Assuntos
Mycobacterium tuberculosis/citologia , Mycobacterium tuberculosis/enzimologia , Proteínas Serina-Treonina Quinases/química , Proteínas Serina-Treonina Quinases/metabolismo , Animais , Membrana Celular/metabolismo , Proliferação de Células , Espaço Extracelular/metabolismo , Regulação Bacteriana da Expressão Gênica , Espaço Intracelular/metabolismo , Camundongos , Viabilidade Microbiana , Mycobacterium tuberculosis/genética , Mycobacterium tuberculosis/fisiologia , Proteínas Serina-Treonina Quinases/genética , Estrutura Terciária de Proteína , Transporte Proteico
10.
Biomed Chromatogr ; 29(10): 1594-603, 2015 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25829259

RESUMO

Artemisinin (AN) and artemisinic acid (AA), valuable phyto-pharmaceutical molecules, are well known anti-malarials, but their activities against diseases like cancer, schistosomiasis, HIV, hepatitis-B and leishmaniasis are also being reported. For the simultaneous estimation of AN and AA in the callus and leaf extracts of A. annua L. plants, we embarked upon a simple, rapid, selective, reliable and fairly economical high performance thin layer chromatography (HPTLC) method. Experimental conditions such as band size, chamber saturation time, migration of solvent front and slit width were critically studied and the optimum conditions were selected. The separations were achieved using toluene-ethyl acetate, 9:1 (v/v) as mobile phase on pre-coated silica gel plates, G 60F254 . Good resolution was achieved with Rf values of 0.35 ± 0.02 and 0.26 ± 0.02 at 536 nm for AN and 626 nm for AA, respectively, in absorption-reflectance mode. The method displayed a linear relationship with r(2) value 0.992 and 0.994 for AN and AA, respectively, in the concentration range of 300-1500 ng for AN and 200-1000 ng for AA. The method was validated for specificity by obtaining in-situ UV overlay spectra and sensitivity by estimating limit of detection (30 ng for AN and 15 ng for AA) and limit of quantitation (80 ng for AN and 45 ng for AA) values. The accuracy was checked by the recovery studies conducted at three different levels with the known concentrations and the average percentage recovery was 101.99% for AN and 103.84% for AA. The precision was analyzed by interday and intraday precision and was 1.09 and 1.00% RSD for AN and 1.22 and 6.05% RSD for AA. The analysis of statistical data substantiates that this HPTLC method can be used for the simultaneous estimation of AN and AA in biological samples.


Assuntos
Artemisininas/análise , Cromatografia em Camada Fina/métodos , Artemisia annua/química , Cromatografia Líquida de Alta Pressão/métodos , Cromatografia em Camada Fina/economia , Limite de Detecção , Reprodutibilidade dos Testes , Sensibilidade e Especificidade , Raios Ultravioleta
11.
J Ayub Med Coll Abbottabad ; 27(3): 680-3, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26721039

RESUMO

BACKGROUND: Performing an episiotomy is generally reserved for complicated childbirths, in cases of foetal distress, or when tearing of tissues with serious consequences are foreseen. In addition to the extent of the trauma, the surgical skill, repair after childbirth can have an important effect on the magnitude and degree of morbidity experienced by women after repair. The best technique for this repair would be that which produces less pain in the short and long term. The study was done with an objective to compare the frequency and severity of pain (slight/severe) by using interrupted and continuous methods for repair of episiotomy or second degree perineal tears. METHODS: It is a randomized control trial. This study was carried out in a Gynaecology and Obstetrics department of Benazir Bhutto Hospital Rawalpindi which is a tertiary care hospital. The duration of study was six months. One hundred & thirty-eight primigravidas (69 in each group) were included in the study. RESULTS: Majority of the patients in both groups belonged to 20-25 years age group, i.e., 48.53% (n=33) in group-A and 50% (n=34) in group-B, mean and SD, was 27.69±3.21 in group-A and 28.16±3.89 in group-B, gestation age of the patients in group-A 77.94% (n=53) and 83.82% (n=57) in group-B between 37-40 weeks of gestation. Complication of pain and its severity in both groups at 24 hours and 10th day were compared which showed no significant difference at any severity (i.e., no pain, mild moderate/severe). CONCLUSION: There is no significant difference in frequency and severity of pain (slight/severe) in using interrupted and continuous methods for repair of second degree perineal tears or episiotomy


Assuntos
Parto Obstétrico/efeitos adversos , Episiotomia/efeitos adversos , Períneo/lesões , Procedimentos de Cirurgia Plástica/métodos , Lesões dos Tecidos Moles/cirurgia , Técnicas de Sutura/instrumentação , Suturas , Adulto , Feminino , Humanos , Períneo/cirurgia , Gravidez , Ruptura/cirurgia , Lesões dos Tecidos Moles/etiologia , Cicatrização , Adulto Jovem
12.
J Ayub Med Coll Abbottabad ; 27(1): 51-4, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26182737

RESUMO

BACKGROUND: Emergency contraception Pills (ECP) provides a safe and effective means of post coital treatment and prevents at least 75% of expected pregnancies resulting from unprotected intercourse. The purpose of the study was to assess the awareness regarding emergency contraception and to see the knowledge attitude and preference about emergency contraception. METHODS: This was a descriptive cross sectional study carried out at Combined Military Hospital (CMH) Khuzdar. A total of 200 married women of reproductive age group who agreed to participate in the study were interviewed using a self-reported comprehensive, structured closed ended questionnaire. RESULTS: 77% of the women were practicing some contraceptive method at the time of study. Most were using condoms for contraception. 16% of all respondents have never used any contraceptive in their life. 70% believe that religion of Islam is not a barrier in family planning. Only 7.5% of the women were aware about ECP. CONCLUSION: Knowledge about ECP is poor among the women of child bearing age. There is a room for improvement regarding the awareness and use of ECP which can contribute to prevention of unwanted pregnancies.


Assuntos
Comportamento Contraceptivo/etnologia , Anticoncepção Pós-Coito/estatística & dados numéricos , Anticoncepcionais Pós-Coito/farmacologia , Emergências , Adulto , Comportamento Contraceptivo/psicologia , Anticoncepção Pós-Coito/ética , Estudos Transversais , Feminino , Humanos , Islamismo , Paquistão/epidemiologia , Gravidez , Prevalência , Inquéritos e Questionários , Adulto Jovem
13.
BMC Microbiol ; 14: 84, 2014 Apr 04.
Artigo em Inglês | MEDLINE | ID: mdl-24708478

RESUMO

BACKGROUND: The absence of rapid tests evaluating antibiotic susceptibility results in the empirical prescription of antibiotics. This can lead to treatment failures due to escalating antibiotic resistance, and also furthers the emergence of drug-resistant bacteria. This study reports a rapid optical method to detect ß-lactamase and thereby assess activity of ß-lactam antibiotics, which could provide an approach for targeted prescription of antibiotics. The methodology is centred on a fluorescence quenching based probe (ß-LEAF--ß-Lactamase Enzyme Activated Fluorophore) that mimics the structure of ß-lactam antibiotics. RESULTS: The ß-LEAF assay was performed for rapid determination of ß-lactamase production and activity of ß-lactam antibiotic (cefazolin) on a panel of Staphylococcus aureus ATCC strains and clinical isolates. Four of the clinical isolates were determined to be lactamase producers, with the capacity to inactivate cefazolin, out of the twenty-five isolates tested. These results were compared against gold standard methods, nitrocefin disk test for ß-lactamase detection and disk diffusion for antibiotic susceptibility, showing results to be largely consistent. Furthermore, in the sub-set of ß-lactamase producers, it was demonstrated and validated that multiple antibiotics (cefazolin, cefoxitin, cefepime) could be assessed simultaneously to predict the antibiotic that would be most active for a given bacterial isolate. CONCLUSIONS: The study establishes the rapid ß-LEAF assay for ß-lactamase detection and prediction of antibiotic activity using S. aureus clinical isolates. Although the focus in the current study is ß-lactamase-based resistance, the overall approach represents a broad diagnostic platform. In the long-term, these studies form the basis for the development of assays utilizing a broader variety of targets, pathogens and drugs.


Assuntos
Antibacterianos/farmacologia , Staphylococcus aureus/efeitos dos fármacos , Staphylococcus aureus/enzimologia , beta-Lactamases/análise , beta-Lactamas/farmacologia , Cefazolina/farmacologia , Fluorometria/métodos , Humanos , Testes de Sensibilidade Microbiana/métodos , Infecções Estafilocócicas/microbiologia , Staphylococcus aureus/isolamento & purificação
14.
Med J Armed Forces India ; 70(4): 344-8, 2014 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-25382908

RESUMO

BACKGROUND: Professional Divers are at a high risk of Sensorineural Hearing Impairment. Divers may sustain sub-clinical hearing loss that is not identified on pure tone audiometry (PTA). Otoacoustic Emissions (OAE) reflect the functional status of the Outer Hair Cells. OAE testing constitutes the only non-invasive means of objective cochlear investigation and may be a more sensitive measure than PTA in identifying sub-clinical hearing loss. METHODS: This cross-sectional study was performed to determine utility of Otoacoustic Emissions testing in detecting sub-clinical Inner Ear damage in divers of Indian Navy. Transient Evoked Otoacoustic Emissions (TEOAE) levels were measured in 50 audiologically asymptomatic ship divers of Indian Navy and compared with control group comprising of 50 normal hearing individuals. RESULTS: No statistically significant differences were observed between the study and control group. Also no correlation was observed between diving years and TEOAE levels. There was no correlation greater than -0.49 between diving years and TEOAE SNR. CONCLUSIONS: We concluded that TEOAE levels are not a sensitive tool in identifying ships divers without any history of noise exposure at risk for developing hearing loss.

15.
Clin Dysmorphol ; 33(2): 63-68, 2024 Apr 01.
Artigo em Inglês | MEDLINE | ID: mdl-38441200

RESUMO

Aniridia is an autosomal dominant condition characterized by the complete or partial absence of the iris, often with additional presentations such as foveal hypoplasia, nystagmus, cataract, glaucoma and other ocular abnormalities. Most cases are caused by heterozygous mutations in the paired box 6 gene (PAX6), which codes for a transcription factor that regulates eye development. Four patients from our hospital who presented with ocular phenotypes were recruited for research sequencing with informed consent. Sanger sequencing of PAX6 coding exons or exome sequencing was performed on genomic DNA from venous blood samples. Variants in PAX6 were identified in the four patients. Two variants are recurrent single-nucleotide substitutions - one is a substitution found in a patient with bilateral aniridia, whereas the other is a splice variant in a patient with nystagmus and neuroblastoma. The other two variants are novel and found in two patients with isolated aniridia. Both are small duplications that are predicted to lead to premature termination. For the recurrent variants, the comparison of phenotypes for patients with identical variants would shed light on the mechanisms of pathogenesis, and the discovery of two novel variants expands the spectrum of PAX6 mutations.


Assuntos
Aniridia , Catarata , Humanos , Face , Aniridia/genética , Catarata/genética , Éxons , Sudeste Asiático , Fator de Transcrição PAX6/genética
16.
Am J Clin Pathol ; 2024 Mar 04.
Artigo em Inglês | MEDLINE | ID: mdl-38437878

RESUMO

OBJECTIVES: Gender equity studies have shown that women are underrepresented in journal editor in chief positions, which confer major professional opportunities and influence. We sought to systematically investigate editor in chief gender and journal attributes within pathology. METHODS: We constructed a journal data set using the Scimago Journal & Country Rank and Clarivate Journal Citation Reports databases. We also included official journals of the major medical societies for the 12 pathology subspecialties recognized by the Association of American Medical Colleges. The final data set included 126 journals. We obtained editor in chief gender, impact factor, publication model (ie, hybrid access vs open access), year of founding, and geographic location for all included pathology journals. RESULTS: Women made up only 18% of the 141 total editor in chief positions. This inequity was present irrespective of all pathology journal variables studied. Among 10 journals with 2 editor in chief positions, 5 had only men and 5 had 1 man and 1 woman. All 3 journals with 3 editor in chief positions had 2 men and 1 woman. CONCLUSIONS: Women are significantly underrepresented among editor in chiefs across pathology journals. Journals and affiliated members should advocate for diversity among these influential positions, given their impact on research, science, and medicine.

17.
Appl Environ Microbiol ; 79(5): 1718-29, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23315736

RESUMO

Escherichia coli-mycobacterium shuttle vectors are important tools for gene expression and gene replacement in mycobacteria. However, most of the currently available vectors are limited in their use because of the lack of extended multiple cloning sites (MCSs) and convenience of appending an epitope tag(s) to the cloned open reading frames (ORFs). Here we report a new series of vectors that allow for the constitutive and regulatable expression of proteins, appended with peptide tag sequences at their N and C termini, respectively. The applicability of these vectors is demonstrated by the constitutive and induced expression of the Mycobacterium tuberculosis pknK gene, coding for protein kinase K, a serine-threonine protein kinase. Furthermore, a suicide plasmid with expanded MCS for creating gene replacements, a plasmid for chromosomal integrations at the commonly used L5 attB site, and a hypoxia-responsive vector, for expression of a gene(s) under hypoxic conditions that mimic latency, have also been created. Additionally, we have created a vector for the coexpression of two proteins controlled by two independent promoters, with each protein being in fusion with a different tag. The shuttle vectors developed in the present study are excellent tools for the analysis of gene function in mycobacteria and are a valuable addition to the existing repertoire of vectors for mycobacterial research.


Assuntos
Vetores Genéticos , Biologia Molecular/métodos , Mycobacterium tuberculosis/genética , Escherichia coli/genética , Expressão Gênica , Marcação de Genes , Dados de Sequência Molecular , Mapeamento de Interação de Proteínas , Recombinação Genética , Análise de Sequência de DNA
18.
Med J Armed Forces India ; 74(4): 386-389, 2018 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-30449929
19.
Environ Technol ; 44(15): 2300-2314, 2023 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-34994296

RESUMO

Low density Polyethylene (LDPE) in various forms has become a part of life. Its accretion due to non degradable nature is concern, endangering life on earth. Amongst various methods of LDPE disposal bioremediation is regarded as ecofriendly & widely accepted. Current investigation was an attempt to isolate potent PE degrading fungus from municipal landfill soils of Bhopal, India loaded with plastic waste.16 fungal isolates were recorded from the site; PE deteriorating fungus was screened using mineral salt agar medium amended with 3% LDPE powder as sole carbon source. The isolate Penicillium citrinum showed fast fungal colony growth in screening medium was selected for biodegradation study. P.citrinum showed 38.82 ± 1.08% weight loss of untreated LDPE pieces; to improve the degradation capacity nitric acid pretreatment was performed; biodegradation was significantly stimulated by 47.22 ± 2.04% after it's pretreatment. Laccase, lipase, esterase & manganese peroxidase activities were assayed by spectrophotometer. LDPE biodegradation was analyzed by weight loss %, change in pH during fungal growth, field emission scanning electron microscopy (FE-SEM), fourier transform infrared spectroscopy (FTIR) & thermogravimetric analysis (TGA). FTIR spectra showed appearance of new functional groups assigned to hydrocarbon biodegradation, confirming enzymatic role in process. Changes in FTIR spectra of LDPE samples (untreated & pretreated) before & after biodegradation & surface changes in the biodegraded LDPE (indicated from FE-SEM) confirmed depolymerization of LDPE. Further changes in thermal decomposition rates of biodegraded samples in comparison to control, validate biodegradation. This is the first report signifying high competence of P.citrinum in LDPE degradation without prior pretreatment.


Assuntos
Penicillium , Polietileno , Polietileno/química , Plásticos , Biodegradação Ambiental , Instalações de Eliminação de Resíduos , Penicillium/metabolismo , Índia
20.
Indian J Otolaryngol Head Neck Surg ; 75(Suppl 1): 83-87, 2023 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-37206739

RESUMO

The most popular objective physiologic test for detecting hearing loss that is in use today is the ABR, however it is not frequency specific. The frequency specific tool available for evaluation of hearing is ASSR. The study is aimed to assess the ability of ASSR to estimate hearing thresholds and identify the ideal modulation frequency in hearing impaired personnel. All subjects and controls were subjected to PTA to determine presence/absence of hearing loss, and the nature and configuration of the hearing loss if any. The subjects were then subjected to ASSR testing to objectively ascertain hearing thresholds. The PTA thresholds obtained and the hearing thresholds obtained by ASSR were correlated in this study. The study was carried out in 100 subjects under the age of 50 years (50 with normal hearing & 50 with impaired hearing by PTA) after obtaining informed consent. Moderate correlation was found between PTA and ASSR thresholds only in certain frequencies while in other frequencies the correlation though present, was low. This study concluded that ASSR system could be used to estimate hearing thresholds only approximately as no significant linear correlations were found between PTA thresholds and ASSR at the tested frequencies.

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