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BACKGROUND: Betulinic acid (BA), a member of pentacyclic triterpenes has shown important biological activities like anti-bacterial, anti-malarial, anti-inflammatory and most interestingly anticancer property. To overcome its poor aqueous solubility and low bioavailability, structural modifications of its functional groups are made to generate novel lead(s) having better efficacy and less toxicity than the parent compound. BA analogue, 2c was found most potent inhibitor of colon cancer cell line, HT-29 cells with IC50 value 14.9 µM which is significantly lower than standard drug 5-fluorouracil as well as parent compound, Betulinic acid. We have studied another mode of PCD, autophagy which is one of the important constituent of cellular catabolic system as well as we also studied proteasomal degradation pathway to investigate whole catabolic pathway after exploration of 2c on HT-29 cells. METHODS: Mechanism of autophagic cell death was studied using fluorescent dye like acridine orange (AO) and monodansylcadaverin (MDC) staining by using fluorescence microscopy. Various autophagic protein expression levels were determined by Western Blotting, qRT-PCR and Immunostaining. Confocal Laser Scanning Microscopy (CLSM) was used to study the colocalization of various autophagic proteins. These were accompanied by formation of autophagic vacuoles as revealed by FACS and transmission electron microscopy (TEM). Proteasomal degradation pathway was studied by proteasome-Glo™ assay systems using luminometer. RESULTS: The formation of autophagic vacuoles in HT-29 cells after 2c treatment was determined by fluorescence staining--confirming the occurrence of autophagy. In addition, 2c was found to alter expression levels of different autophagic proteins like Beclin-1, Atg 5, Atg 7, Atg 5-Atg 12, LC3B and autophagic adapter protein, p62. Furthermore we found the formation of autophagolysosome by colocalization of LAMP-1 with LC3B, LC3B with Lysosome, p62 with lysosome. Finally, as proteasomal degradation pathway downregulated after 2c treatment colocalization of ubiquitin with lysosome and LC3B with p62 was studied to confirm that protein degradation in autophagy induced HT-29 cells follows autolysosomal pathway. CONCLUSIONS: In summary, betulinic acid analogue, 2c was able to induce autophagy in HT-29 cells and as proteasomal degradation pathway downregulated after 2c treatment so protein degradation in autophagy induced HT-29 cells follows autolysosomal pathway.
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Autofagia/efeitos dos fármacos , Neoplasias do Colo/tratamento farmacológico , Complexo de Endopeptidases do Proteassoma/efeitos dos fármacos , Triterpenos/administração & dosagem , Neoplasias do Colo/genética , Neoplasias do Colo/patologia , Células HT29 , Humanos , Proteínas de Neoplasias/biossíntese , Triterpenos Pentacíclicos , Complexo de Endopeptidases do Proteassoma/genética , Proteólise/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Triterpenos/química , Vacúolos/efeitos dos fármacos , Vacúolos/patologia , Ácido BetulínicoRESUMO
Knowing the spectrum, prevalence, and modes of diagnosis of pulmonary aspergillosis (PA) will be beneficial to clinicians for its early diagnosis and management. This study aims to estimate the prevalence, spectrum, and role of serological tests and radiological findings in the diagnosis of PA. A total of 150 patients were suspected of having PA after obtaining relevant clinical history and radiological imaging. The patients were grouped into each spectrum of PA as invasive PA (IPA), chronic necrotizing PA (CNPA), aspergilloma, allergic bronchopulmonary aspergillosis (ABPA) based on predisposing factors, clinical and radiological findings, and the guidelines of the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG). Samples (bronchoalveolar lavage (BAL), sputum, blood) were collected from these patients and processed in a microbiology lab. BAL and sputum were subjected to microscopy by potassium hydroxide mount, calcofluor white mount, and culture. The serum was separated from blood by centrifugation and subjected to specific serological tests based on the spectrum of PA that the patient was suspected to have. For IPA, serum and BAL galactomannan antigen enzyme-linked immunosorbent assay (ELISA) was performed. For CNPA and aspergilloma, the anti-Aspergillus IgG antibody ELISA was performed. For ABPA, the tests performed were total immunoglobulin E (IgE) ELISA, Aspergillus fumigatus-specific IgE ELISA, and anti-Aspergillus immunoglobulin G (IgG) antibody ELISA. After compiling the clinical, radiological, culture, and serological findings, patients were diagnosed to have a particular spectrum of PA. The prevalence of IPA was 1.4%, CNPA was 4%, ABPA was 3.2%, and aspergilloma was 2.9%. CNPA was the predominant spectrum of PA in our study. Culture positivity for Aspergillus species was seen the highest in aspergilloma patients, followed by IPA, ABPA, and CNPA patients. A. fumigatus was the most common causative agent of PA, except for IPA for which Aspergillus flavus was the most common causative. Aspergillus niger and Aspergillus terreus were less the frequent causes of PA. A combination of radiological, microbiological, and serological tests along with clinical correlation is needed to confirm the diagnosis of PA.
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BACKGROUND: Coagulase-negative staphylococci (CoNS) are one of the frequently isolated bacteria from blood cultures. Since they are part of the normal skin flora, they were previously considered contaminants. But now, they can be considered as established pathogens causing bloodstream infection (BSI). This study aims to estimate the prevalence of CoNS in BSI cases. METHODS: This study was conducted at the Microbiology Department, All India Institute of Medical Sciences (AIIMS), Raipur, India, for eight months (January 2022 to August 2022). Data were collected retrospectively from medical and laboratory records. Paired blood cultures from 5085 clinically suspected sepsis cases were subjected to aerobic culture for five days in the BacT ALERT 3D system. Pathogenicity was established after recovery of CoNS from paired blood cultures of symptomatic patients. RESULTS: CoNS were isolated from 2.35% of patients, the most common species being Staphylococcus haemolyticus (51.67%). About 90% of isolates were methicillin-resistant. All the isolates were susceptible to linezolid, teicoplanin, and vancomycin, except one isolate of S. haemolyticus which was intermediate to vancomycin. Minimum inhibitory concentration (MIC) 50 and MIC 90 for vancomycin were 1 ug/ml and 2 ug/ml, respectively. Conclusion: Paired blood cultures are necessary to determine the pathogenicity of CoNS in BSI cases. A high prevalence of methicillin resistance, accompanied by high resistance rates to other non-beta lactam antibiotics, warrants the strict implementation of antimicrobial stewardship practices.
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INTRODUCTION: The current systematic review aimed to collect and analyze the comprehensive evidence regarding Polymerase Spiral Reaction (PSR) and to estimate its diagnostic performance and usefulness as a point-of-care (PoC) assay. METHODS: Literature was retrieved systematically from 2015 to 2023 from PubMed and Scopus. Studies were screened and selected against pre-determined inclusion and exclusion criteria. Quality assessment and risk of bias were critiqued using QUADAS-2. A systematic, qualitative narrative synthesis was employed to synthesize the data. RESULTS: 11 studies were selected for the systematic review, testing diseases in humans utilizing PSR. Only 2 studies clinically validated the test with a sample size > 150. 5 studies were of poor quality; 3 studies were of moderate quality and 3 studies were deemed to be of high quality. 3 studies quantified the diagnostic throughput and reported clinical sensitivity and specificity of PSR approaching to be > 92% and ~ 100%, respectively. CONCLUSION: Polymerase spiral reaction promises to be an optimistic isothermal assay; however, a huge research gap can be attributed to the lack of statistical and clinical evidence to validate the assay. Adequate research, focused on optimization, coupled with statistical and clinical validation, can help in estimating its true diagnostic potential and applicability. REGISTRATION AND PROTOCOL: A detailed protocol of this review is registered and available in Prospero (registration number CRD42023406265).
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Técnicas de Diagnóstico Molecular , Sistemas Automatizados de Assistência Junto ao Leito , Testes Imediatos , Humanos , Técnicas de Diagnóstico Molecular/métodos , Técnicas de Diagnóstico Molecular/normas , Sensibilidade e Especificidade , Técnicas de Amplificação de Ácido Nucleico/métodosRESUMO
OBJECTIVES: The purpose of this study was to determine the value of faecal calprotectin (f-CP) in distinguishing between bacterial and viral aetiologies of infective diarrhoea in children attending a tertiary care hospital in Central India. METHODS: Stool samples from children aged 3 months to 10 years who had acute or persistent diarrhoea were processed for microscopy, bacterial culture, and viral antigen detection (Rotavirus and Norovirus). The remaining samples, as well as stool samples from 20 healthy controls, were tested for f-CP using the enzyme linked immunosorbent assay. RESULTS: Among 48 patients, 21 (43.7%) had bacterial diarrhoea, 14 (29.2%) had viral diarrhoea, and 13 (27.1%) had an unidentified aetiology. The median f-CP values were significantly (p â= â0.004) higher in children with bacterial diarrhoea (75.2 âµg/g; IQR-18.75-239.15) than in children with viral diarrhoea (75.2 âµg/g; IQR-123.5-1987.5). Bacterial aetiology could be reliably predicted at the optimum f-CP concentrations of >541 âµg/g and >238.4 âµg/g in children aged 1 and 1-4 years, with an area under the curve of 0.767 and 0.867, respectively, using receiver-operator characteristic analysis. CONCLUSIONS: Faecal calprotectin could reliably distinguish between bacterial and viral aetiologies of diarrhoea in children aged up to four years, but at relatively higher age-specified cut off values.
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Infecções Bacterianas , Infecções por Enterovirus , Rotavirus , Criança , Humanos , Complexo Antígeno L1 Leucocitário/análise , Diarreia/diagnóstico , Diarreia/microbiologia , Ensaio de Imunoadsorção Enzimática , Fezes/microbiologia , BiomarcadoresRESUMO
BACKGROUND: Healthcare-associated infections (HAIs) are one of the most common adverse events in patient care that account for substantial morbidity and mortality. We evaluate the existing Infection Prevention and Control (IPC) practices in hospitals participating in the nationally representative HAI Surveillance network. METHODS: This cross-sectional survey was conducted in 23 hospitals across 22 states of India from October-2015 to September-2018 in the HAI surveillance network. The World Health Organization (WHO) IPC core components assessment tool for health-care facility level (IPCAT-H) was adapted from IPC assessment tool developed by US Centers for Disease Control and Prevention (US CDC) under the Epidemiology and Laboratory Capacity (ELC) Infection Control Assessment and Response (ICAR) Program. Mann-Whitney U test was used to calculate the significant difference between scores (P < .05). RESULTS: Amongst the participating hospitals, 7 were private sectors and 16 were public health care facilities. Infection IPCAT-H average score per multimodal strategy was less than 50% for programmed IPC activities (45.7); implementation of health care workers (HCWs) immunization programme (43.5%); monitoring and evaluation component (38.30%). CONCLUSIONS: There is potential for improvement in Human Resources, Surveillance of HAIs as well as Monitoring and Evaluation components.
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Infecção Hospitalar , Controle de Infecções , Humanos , Controle de Infecções/métodos , Autorrelato , Estudos Transversais , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/prevenção & controle , HospitaisRESUMO
The rapid emergence of drug resistance in Acinetobacter baumannii has put forward the use of colistin as a last-resort treatment for infections with A. baumannii. Empirical colistin use without prior susceptibility testing has been one of the factors that has been promoting drug resistance in low-resource settings. In this regard, while the advocated broth microdilution (BMD) method for colistin susceptibility testing is often considered cumbersome, the preferable colistin broth disk elution (CBDE) method has not yet been approved for A. baumannii. To prevent the underreporting of colistin susceptibility, we tested the CBDE method for A. baumannii and compared the results with those of BMD. A total of 125 A. baumannii, including 100 susceptible and 25 resistant isolates were tested via the CBDE method and compared with the standard BMD method. The essential agreement, categorical agreement, sensitivity, and specificity for CBDE were 97.6% (n = 122), 98.4% (n = 123), 100%, and 98.40%, respectively. The percentage of major error found was 1.6% (n = 2), and no very major error was found. CBDE in A. baumannii could be considered in low-resource settings. IMPORTANCE The relatively cumbersome broth microdilution (BMD) method for routine colistin susceptibility testing has not been adopted, especially in low-resource settings, often leading to the underreporting of colistin susceptibility and the promotion of the empirical use of colistin. In this regard, the much-preferred colistin broth disk elution (CBDE) method has not yet been approved for A. baumannii. We evaluated colistin susceptibility via the CBDE method, compared the results with those of the BMD method in 125 A. baumannii isolates with various profiles, and inferred that the CBDE method using 50 µL inoculum could be helpful, at least in resource-limited setups, versus not reporting susceptibility testing for colistin.
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Acinetobacter baumannii , Colistina , Colistina/farmacologia , Testes de Sensibilidade Microbiana , Antibacterianos/farmacologia , Farmacorresistência Bacteriana MúltiplaRESUMO
PURPOSE: Since ancient era leprosy is existing across the world. India, Indonesia and Brazil still harbour major proportion of global cases. Child leprosy and Grade II disability indicate delayed diagnosis and persistence of transmission in community. So, this study was conducted with aim to evaluate the diagnostic efficacy of PCR in comparison to SSS (Slit Skin Smear) microscopy for detection of leprosy in early stages in both cases and carriers (contacts). METHODS: A cross sectional observational study was conducted on 100 subjects including 50 clinically diagnosed new cases of leprosy and their 50 contacts. Each group was subjected to SSS (Slit Skin Smear) microscopy and PCR using RLEP gene as target. RESULTS: The overall male: female ratio was 2.44. The Slit Skin smear (SSS) microscopy positivity was 34% (nâ¯=â¯17/50) among cases while it was 0% (nâ¯=â¯0/50) among contacts. The overall positivity for PCR was 42% (nâ¯=â¯42/100) being 66% (nâ¯=â¯33/50) in cases and 18% (nâ¯=â¯9/50) in contacts. About 30% (nâ¯=â¯25/83) of all the microscopically negative subjects were found to be positive by PCR. CONCLUSIONS: PCR was found to be a better diagnostic tool both among cases and their contacts. It should be used for screening contacts for early diagnosis and treatment and thus preventing transmission in community. KEY MESSAGE: To diagnose case and contacts of leprosy in early stages even in very low bacterial density using PCR.
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Hanseníase , Mycobacterium leprae , Criança , Estudos Transversais , Feminino , Hospitais de Ensino , Humanos , Hanseníase/diagnóstico , Hanseníase/microbiologia , Masculino , Mycobacterium leprae/genética , Reação em Cadeia da Polimerase , Atenção Terciária à SaúdeRESUMO
BACKGROUND: Healthcare workers (HCWs) stand the risk of acquiring infection directly, while attending to patients or indirectly while handling and testing patient specimens. Considering this, the present study was planned to assess Personal Protective Equipment (PPE) breaches and exposures among HCWs working in COVID-19 wards/ screening areas and to evaluate their COVID-19 positivity rates post-exposure concerning the level of exposure, type of PPE breach, and the cadre of HCWs exposed in COVID-19 wards. METHODS: This retrospective cross-sectional study involved the analysis of all instances of PPE breaches which occurred during a period of nine months from June 2020 to February 2021 at a tertiary care level hospital in Central India. The analysis included all exposures involving any cadre of HCWs that occurred while handling the patients or while doffing the contaminated PPE in COVID -19 wards. RESULTS: A total of 347 PPE breaches were analyzed from the available records of the Hospital Infection Control team repository. Amongst the 347 breaches, 268 (77.2%) were classified as low-risk exposures and 79 (22.8%) as high-risk exposures. Cadre wise distribution of high and low-risk exposures revealed that, PPE breaches occurred most commonly in the category of nursing officers (n = 174, 50.1%). Among all of the breaches, 15.2% of high-risk exposures and 2.6% of low-risk exposures resulted in COVID-19 positivity with a cumulative positivity of 5.4%. Collectively, non-mask related breaches accounted for the majority (63.2%) of the positive COVID-19 cases. CONCLUSION: Appropriate use of PPE by HCWs is vital for their protection. However, breaches in the use of PPE may occur while managing COVID-19 patients due to physical and mental exhaustion among HCWs resulting from work overload. Early identification and appropriate management of HCWs with high-risk exposures can help prevent transmission to other hospital staff and patients, thus preserving resources and workforce.
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COVID-19 , Equipamento de Proteção Individual , COVID-19/epidemiologia , COVID-19/prevenção & controle , Estudos Transversais , Pessoal de Saúde , Humanos , Transmissão de Doença Infecciosa do Paciente para o Profissional/prevenção & controle , Recursos Humanos em Hospital , Estudos RetrospectivosRESUMO
Melioidosis is a potentially fatal, life-threatening infection caused by the gram-negative saprophytic organism Burkholderia. It is a disease endemic to Southeast Asia and Northern Australia. This infection transmits through direct contact, cutaneous inoculation, inhalation, or ingestion, and patients clinically exhibit abscesses in single or multiple organs. It is clinically under-reported due to a low index of suspicion, lack of diagnostic facilities, and misdiagnosis as tuberculosis. Infections of the musculoskeletal system are exceedingly rare, and clinical presentation may vary from the involvement of femoral bone, palmar tenosynovitis, and parietal bone osteomyelitis secondary to central nervous system involvement. The rarity of the melioidosis to secondarily infect a developmental odontogenic cyst leading to focal osteomyelitis of mandible prompts the clinician toward thorough evaluation for early diagnosis and treatment.
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BACKGROUND: Health-care-associated infections (HAIs) cause significant morbidity and mortality globally, including in low-income and middle-income countries (LMICs). Networks of hospitals implementing standardised HAI surveillance can provide valuable data on HAI burden, and identify and monitor HAI prevention gaps. Hospitals in many LMICs use HAI case definitions developed for higher-resourced settings, which require human resources and laboratory and imaging tests that are often not available. METHODS: A network of 26 tertiary-level hospitals in India was created to implement HAI surveillance and prevention activities. Existing HAI case definitions were modified to facilitate standardised, resource-appropriate surveillance across hospitals. Hospitals identified health-care-associated bloodstream infections and urinary tract infections (UTIs) and reported clinical and microbiological data to the network for analysis. FINDINGS: 26 network hospitals reported 2622 health-care-associated bloodstream infections and 737 health-care-associated UTIs from 89 intensive care units (ICUs) between May 1, 2017, and Oct 31, 2018. Central line-associated bloodstream infection rates were highest in neonatal ICUs (>20 per 1000 central line days). Catheter-associated UTI rates were highest in paediatric medical ICUs (4·5 per 1000 urinary catheter days). Klebsiella spp (24·8%) were the most frequent organism in bloodstream infections and Candida spp (29·4%) in UTIs. Carbapenem resistance was common in Gram-negative infections, occurring in 72% of bloodstream infections and 76% of UTIs caused by Klebsiella spp, 77% of bloodstream infections and 76% of UTIs caused by Acinetobacter spp, and 64% of bloodstream infections and 72% of UTIs caused by Pseudomonas spp. INTERPRETATION: The first standardised HAI surveillance network in India has succeeded in implementing locally adapted and context-appropriate protocols consistently across hospitals and has been able to identify a large number of HAIs. Network data show high HAI and antimicrobial resistance rates in tertiary hospitals, showing the importance of implementing multimodal HAI prevention and antimicrobial resistance containment strategies. FUNDING: US Centers for Disease Control and Prevention cooperative agreement with All India Institute of Medical Sciences, New Delhi. TRANSLATION: For the Hindi translation of the abstract see Supplementary Materials section.
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Anti-Infecciosos , Infecção Hospitalar , Pneumonia Associada à Ventilação Mecânica , Sepse , Infecções Urinárias , Criança , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/prevenção & controle , Humanos , Recém-Nascido , Klebsiella , Pneumonia Associada à Ventilação Mecânica/complicações , Pneumonia Associada à Ventilação Mecânica/epidemiologia , Pneumonia Associada à Ventilação Mecânica/microbiologia , Estudos Prospectivos , Centros de Atenção Terciária , Infecções Urinárias/epidemiologiaRESUMO
BACKGROUND: Healthcare associated infections (HAIs) are prevalent and difficult to treat worldwide. Most HAIs can be prevented by effective implementation of Infection Prevention and Control (IPC) measures. A survey was conducted to assess the existing IPC practices across a network of Indian Hospitals using the World Health Organization designed self-assessment IPC Assessment Framework (IPCAF) tool. METHODS: This was a cross sectional observation study. Thirty-two tertiary care public and private facilities, part of the existing Indian HAI surveillance network was included. Data collected was analyzed by a central team at All India Institute of Medical Sciences, New Delhi, a tertiary care hospital of India. The WHO questionnaire tool was used to understand the capacity and efforts to implement IPC practices across the network. RESULTS: The overall median score of IPCAF across the network was 620. Based on the final IPCAF score of the facilities; 13% hospitals had basic IPC practices, 28% hospitals had intermediate and 59% hospitals had advanced IPC practices. The component multimodal strategies had the broadest range of score while the component IPC guidelines had the narrowest one. CONCLUSIONS: Quality improvement training for IPC nurses and healthcare professionals are needed to be provided to health facilities.
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Infecção Hospitalar , Controle de Infecções , Infecção Hospitalar/epidemiologia , Infecção Hospitalar/prevenção & controle , Estudos Transversais , Atenção à Saúde , Instalações de Saúde , Humanos , Autorrelato , Inquéritos e QuestionáriosRESUMO
Nitric oxide (NO) has been demonstrated to be a principal effector molecule responsible for mediating intracellular killing of Leishmania parasites, the causative organism of leishmaniasis. As measurement of intracellular NO remains a challenge to biologists, we have developed a flow cytometric approach to perform real time biological detection of NO within Leishmania parasites and parasitized macrophages using a membrane permeable derivative of diaminofluorescein [4,5-diaminofluorescein diacetate (DAF-2DA)]. Initially, assay optimization was performed in Leishmania donovani promastigotes, assay specificity being confirmed using both a NO donor [S-nitroso-N-acetyl-penicillamine (SNAP)] and a NO scavenger [2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazoline-1-oxyl-3-oxide, C-PTIO]. Using 40 µM DAF-2DA, basal levels of intracellular NO were measured which varied in different Leishmania species; addition of conventional anti-leishmanial drugs, antimony and miltefosine translated into a dramatic increase in DAF-2T fluorescence. Furthermore, the assay also measured levels of NO in macrophages, but needed a 20 fold lower concentration of DAF-2DA, being 2 µM. Following parasitization, levels of NO decreased which was normalized following treatment with anti-leishmanial drugs. Similarly monocytes of patients with visceral leishmaniasis at disease presentation showed decreased levels of NO which too reverted on completion of treatment. Taken together, this study opens new perspectives of research regarding monocyte function and provides a real time approach for monitoring the effect of anti-leishmanial compounds.
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Leishmaniose Visceral/patologia , Macrófagos Peritoneais/metabolismo , Óxido Nítrico/biossíntese , Adolescente , Adulto , Idoso , Animais , Tartarato de Antimônio e Potássio/farmacologia , Antiprotozoários/farmacologia , Benzoatos/farmacologia , Feminino , Citometria de Fluxo , Fluoresceína , Humanos , Imidazóis/farmacologia , Técnicas In Vitro , Leishmania/efeitos dos fármacos , Leishmania/metabolismo , Leishmaniose Visceral/metabolismo , Leishmaniose Visceral/parasitologia , Macrófagos Peritoneais/efeitos dos fármacos , Macrófagos Peritoneais/parasitologia , Masculino , Camundongos , Camundongos Endogâmicos BALB C , Pessoa de Meia-Idade , Monócitos/metabolismo , Fosforilcolina/análogos & derivados , Fosforilcolina/farmacologia , Especificidade da Espécie , Adulto JovemRESUMO
2,2'-Diphenyl-3,3'-diindolylethylene (DPDIE) derivatives 3a-g were regioselectively prepared in one pot from indoles 1a-g in the presence of Lewis acids and were subsequently evaluated for cytotoxic activity against human leukemic cell lines, U937 and K562. The most potent compound 3g exhibited IC(50) of 13.0-17.0 µM.
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Antineoplásicos/síntese química , Antineoplásicos/toxicidade , Etilenos/síntese química , Etilenos/farmacologia , Indóis/química , Antineoplásicos/química , Linhagem Celular Tumoral , Sobrevivência Celular/efeitos dos fármacos , Cristalografia por Raios X , Etilenos/química , Citometria de Fluxo , Humanos , Concentração Inibidora 50 , Estrutura MolecularRESUMO
Elizabethkingia meningoseptica is a recognized cause of neonatal meningitis with high mortality rate of approximately 57%, but clinical data detailing these infections remain limited from India. Though this bacteria has a Gram-negative character, it is usually multidrug resistant to antibiotics usually prescribed for Gram-negative bacterial infections and susceptible to antibiotics for Gram-positive bacteria, thus poses a serious challenge to the treating clinicians. Such cases of neonatal meningitis is most commonly associated with prematurity with birth weight < 2,500 g, but here we report an uncommon case of neonatal meningitis due to this rare pathogen in a full-term neonate with weight as per gestational age. The isolate was multidrug resistant and discrepancy was seen between disc diffusion and automated antibiotic susceptibility testing for few antibiotics. The case was successfully managed by treatment with combination of piperacillin-tazobactam, vancomycin, chloramphenicol, and rifampicin for a total duration of 28 days, due to prompt identification of the causative organism and initiation of appropriate antimicrobial therapy early. E. meningoseptica can cause severe infection, with risk of high mortality and neurological sequelae in neonates. Intensive care and multidisciplinary interventions are crucial for case management.
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A fifty-nine-year-old female with corneal ulcer with a history of trauma in the past having clinical presentation of fungal keratitis visited the Ophthalmology outpatient department with a history of redness, watering, pain and white discoloration of the right eye cornea for two months. Sample was cultured on non-nutrient agar medium and revealed Acanthamoeba. Patient was managed with Chlorhexidine eye drop and keratoplasty.
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Bacterial keratitis is a serious, potentially sight-threatening complication in neglected cases of corneal trauma. Few bacteria and fungi are implicated in the pathogenesis of bacterial keratitis after trauma; however, keratitis by an opportunistic organism is rare. We report here a case of keratitis caused by Corynebacterium jeikeium (C. jeikeium) after accidental trauma with rice grains. The patient presented to us with chronic keratitis, which responded to 5% fortified vancomycin eye drops after microbiological investigation and drug sensitivity.
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Human papilloma virus genotype 16 (HPV-16), a predominant etiological cause of cervical cancer (CC) vary in inflicting oncogenicity according to their geographical distribution and mutational changes. With no published data from central India, the present study aimed to genetically analyze HPV-16 E6/E7 variant obtained from CC women of Chhattisgarh. In twenty one CC patients, PCR amplified E6/E7 genes were decoded by DNA sequencing to study phylogenetic relatedness, mutational changes and their in-silico effect on protein structure. E6 analysis revealed nineteen sequences exhibited intratypic variation. L83V mutation was observed in 76.2% sequences followed by S71C seen in 28.6% sequences. Mutations of E41G, A46G, F47V, R77S, L99V and Q107K were observed in three sequences each. C140 Stop codon mutation has caused early truncation of E6 in three sequences to produce the conformational structural change. In contrast, E7 was relatively more conserved showing D4E (4.7%), G88R (23.8%), I93T (9.5%) and C94S (9.5%) mutations. Other than L83V and S71C, E6 and E7 mutations were reported for the first time from India. E6/E7 nonsynonmous mutations have a spectrum of biological effect in progression of CC. Phylogenetic analysis revealed ten sequence belonged to Asian while eleven to European sublineage to show CC cases in Chhattisgarh are a mix of Asian and European lineage. Asian sequences showing higher frequency of L83V mutations and exclusive presence of S71C and C140 Stop codon mutations may be linked with higher oncogenicity. Various E6/E7 mutational data may prove useful for development of better diagnostic and vaccine for the region of Chhattisgarh.
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[This corrects the article DOI: 10.1371/journal.pone.0249408.].
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BACKGROUND: An appropriate specimen is of paramount importance in Real Time reverse transcription-polymerase chain reaction (rRT-PCR) based diagnosis of novel coronavirus (nCoV) disease (COVID-19). Thus, it's pertinent to evaluate various diversified clinical specimens' diagnostic utility in both diagnosis and follow-up of COVID-19. METHODS: A total of 924 initial specimens from 130 COVID-19 symptomatic cases before initiation of treatment and 665 follow up specimens from 15 randomly selected cases comprising of equal number of nasopharyngeal swab (NPS), oropharyngeal swab (OPS), combined NPS and OPS (Combined swab), sputum, plasma, serum and urine were evaluated by rRT-PCR. RESULTS: Demographic analysis showed males (86) twice more affected by COVID-19 than females (44) (p = 0.00001). Combined swabs showed a positivity rate of 100% followed by NPS (91.5%), OPS (72.3%), sputum (63%), while nCoV was found undetected in urine, plasma and serum specimens. The lowest cycle threshold (Ct) values of targeted genes E, ORF1b and RdRP are 10.56, 10.14 and 12.26 respectively and their lowest average Ct values were found in combined swab which indicates high viral load in combined swab among all other specimen types. Analysis of 665 follow-up multi-varied specimens also showed combined swab as the last specimen among all specimen types to become negative, after an average 6.6 (range 4-10) days post-treatment, having lowest (15.48) and average (29.96) Ct values of ORF1b respectively indicating posterior nasopharyngeal tract as primary nCoV afflicted site with high viral load. CONCLUSION: The combined swab may be recommended as a more appropriate specimen for both diagnosis and monitoring of COVID-19 treatment by rRT-PCR for assessing virus clearance to help physicians in taking evidence-based decision before discharging patients. Implementing combined swabs globally will definitely help in management and control of the pandemic, as it is the need of the hour.