SPECIATION AND ANTIBIOGRAM PATTERN OF TRIBE PROTEEAE WITH SPECIAL REFERENCE TO EXTENDED SPECRUM BETA LACTAMASE (ESBL) AND CARBAPENEMASE DETECTION

Background:Proteusis a common uropathogen causing urinary tract infections in catheterised patients and those with urinary tract abnormalities. It may also lead to pyelonephritis, renal stones & bacteraemia. Multidrug resistant Proteeae isolates are major problem in treating nosocomial infections. Beta lactamase production is being increasingly demonstrated in most of the Proteus species. Apart from ESBL, Carbapenemase production is also emerging, thereby limiting the treatment options. Aim: To isolate, speciate and study the antibiotic resistance pattern of the Proteeae isolates. To identify the extended spectrum beta lactamase andcarbapenemase producing strains of Proteeae isolates by employing phenotypic methods. Materials and Methods: A total of 145 isolates of Proteus species isolated from different clinical samples- urine, pus and sputum, were included in this study. Antibiotic sensitivity testing was performed by Kirby-Bauer disk diffusion method. Screening tests for ESBL & Carbapenemase production was confirmed by Disk diffusion method and Modified Hodge test respectively. Results: Out of the 145 Proteeae isolates, from various clinical samples 70 were from wound swabs, 62 from urine, and 13 from respiratory specimens. The species were identified as 64 P.mirabilis, 48 P. vulgaris, 19 M. morganii, 5 Prov. stuartii and 9 Prov. rettgeri. Different antibiotic resistance patterns were observed in different species. Providencia species showed resistance to most of the antibiotics than the Proteus species and M morganii. Of the total, 52 (36%) were ESBL producers. Among the ESBL producers 6 (11.5%) were Carbapenamase producers. Conclusion: The increasing incidence of multi drug resistant strains in Tribe Proteeae has made antimicrobial susceptibility testing more important. Avoidance of indiscriminate use of antibiotics is the first step in prevention of newly emerging drug resistant strains. KEYWORDS: Proteeae; ESBL; Carbapenemase; Double disk Synergy test; Modified Hodge test

SPECIATION AND ANTIBIOGRAM PATTERN OF TRIBE PROTEEAE WITH SPECIAL REFERENCE TO EXTENDED SPECRUM BETA LACTAMASE (ESBL) AND CARBAPENEMASE DETECTION

Background:Proteusis a common uropathogen causing urinary tract infections in catheterised patients and those with urinary tract abnormalities. It may also lead to pyelonephritis, renal stones & bacteraemia. Multidrug resistant Proteeae isolates are major problem in treating nosocomial infections. Beta lactamase production is being increasingly demonstrated in most of the Proteus species. Apart from ESBL, Carbapenemase production is also emerging, thereby limiting the treatment options. Aim: To isolate, speciate and study the antibiotic resistance pattern of the Proteeae isolates. To identify the extended spectrum beta lactamase andcarbapenemase producing strains of Proteeae isolates by employing phenotypic methods. Materials and Methods: A total of 145 isolates of Proteus species isolated from different clinical samples- urine, pus and sputum, were included in this study. Antibiotic sensitivity testing was performed by Kirby-Bauer disk diffusion method. Screening tests for ESBL & Carbapenemase production was confirmed by Disk diffusion method and Modified Hodge test respectively. Results: Out of the 145 Proteeae isolates, from various clinical samples 70 were from wound swabs, 62 from urine, and 13 from respiratory specimens. The species were identified as 64 P.mirabilis, 48 P. vulgaris, 19 M. morganii, 5 Prov. stuartii and 9 Prov. rettgeri. Different antibiotic resistance patterns were observed in different species. Providencia species showed resistance to most of the antibiotics than the Proteus species and M morganii. Of the total, 52 (36%) were ESBL producers. Among the ESBL producers 6 (11.5%) were Carbapenamase producers. Conclusion: The increasing incidence of multi drug resistant strains in Tribe Proteeae has made antimicrobial susceptibility testing more important. Avoidance of indiscriminate use of antibiotics is the first step in prevention of newly emerging drug resistant strains. KEYWORDS: Proteeae; ESBL; Carbapenemase; Double disk Synergy test; Modified Hodge test

B-cell development : one problem, multiple solutions.

Interspecies variations in the processes of B-cell development and repertoire generation contrast with the greater consistency of T-cell development. B-cell development in mice and humans, with postnatal B-cell generation of new repertoire in the bone marrow throughout life, is regarded as the 'standard' pattern. In contrast, accounts of B cells in birds, sheep, cattle, rabbits and pigs (the 'other' species) describe cessation of gene diversification in the perinatal period, with the gut-associated lymphoid tissue (GALT) functioning as the primary lymphoid organ thereafter. It has become customary to regard the developmental pathways of T and B cells within any individual species as being as dissimilar as the functions of the two mature cell types. Reinterpretation of B-cell development patterns in different species is overdue in response to two types of reports. The first of these describe T-B 'crossover', specifically the intrathymic production of B cells and the extrathymic production of T cells. The second attests to the extent of sharing of B-cell developmental features across the two groups of species. We propose that, as is a feature of other haematopoietic cells, a menu of alternative B- and T-cell pathways has been retained and shared across species. A single pathway usually predominates in any species, masking alternatives. The observed predominance of any pathway is determined by factors such as placental permeability, extent of maturation of the immune system by birth and the feasibility of direct experimental intervention in development

Comparison of the marginal adaptation of direct and indirect composite inlay restorations with optical coherence tomography

OBJECTIVE: The purpose of the study was to use the photonic imaging modality of optical coherence tomography (OCT) to compare the marginal adaptation of composite inlays fabricated by direct and indirect techniques. MATERIAL AND METHODS: Class II cavities were prepared on 34 extracted human molar teeth. The cavities were randomly divided into two groups according to the inlay fabrication technique. The first group was directly restored on cavities with a composite (Esthet X HD, Dentsply, Germany) after isolating. The second group was indirectly restored with the same composite material. Marginal adaptations were scanned before cementation with an invisible infrared light beam of OCT (Thorlabs), allowing measurement in 200 µm intervals. Restorations were cemented with a self-adhesive cement resin (SmartCem2, Dentsply), and then marginal adaptations were again measured with OCT. Mean values were statistically compared by using independent-samples t-test and paired samples t-test (p<0.05), before and after cementation. RESULTS: Direct inlays presented statistically smaller marginal discrepancy values than indirect inlays, before (p=0.00001442) and after (p=0.00001466) cementation. Marginal discrepancy values were increased for all restorations after cementation (p=0.00008839, p=0.000000952 for direct and indirect inlays, respectively). The mean marginal discrepancy value of the direct group increased from 56.88±20.04 µm to 91.88±31.7 µm, whereas the indirect group increased from 107.54±35.63 µm to 170.29±54.83 µm. Different techniques are available to detect marginal adaptation of restorations, but the OCT system can give quantitative information about resin cement thickness and its interaction between tooth and restoration in a nondestructive manner. CONCLUSIONS: Direct inlays presented smaller marginal discrepancy than indirect inlays. The marginal discrepancy values were increased for all restorations that refer to cement thickness after cementation

Identification of Prophages in Bacterial Genomes by Dinucleotide Relative Abundance Difference

BACKGROUND: Prophages are integrated viral forms in bacterial genomes that have been found to contribute to interstrain genetic variability. Many virulence-associated genes are reported to be prophage encoded. Present computational methods to detect prophages are either by identifying possible essential proteins such as integrases or by an extension of this technique, which involves identifying a region containing proteins similar to those occurring in prophages. These methods suffer due to the problem of low sequence similarity at the protein level, which suggests that a nucleotide based approach could be useful. METHODOLOGY: Earlier dinucleotide relative abundance (DRA) have been used to identify regions, which deviate from the neighborhood areas, in genomes. We have used the difference in the dinucleotide relative abundance (DRAD) between the bacterial and prophage DNA to aid location of DNA stretches that could be of prophage origin in bacterial genomes. Prophage sequences which deviate from bacterial regions in their dinucleotide frequencies are detected by scanning bacterial genome sequences. The method was validated using a subset of genomes with prophage data from literature reports. A web interface for prophage scan based on this method is available at http://bicmku.in:8082/prophagedb/dra.html. Two hundred bacterial genomes which do not have annotated prophages have been scanned for prophage regions using this method. CONCLUSIONS: The relative dinucleotide distribution difference helps detect prophage regions in genome sequences. The usefulness of this method is seen in the identification of 461 highly probable loci pertaining to prophages which have not been annotated so earlier. This work emphasizes the need to extend the efforts to detect and annotate prophage elements in genome sequences

The growth inhibitory potential and antimetastatic effect of camel urine on breast cancer cells in vitro and in vivo

Although it may sound unpleasant, camel urine has been consumed extensively for years in the Middle East as it is believed to be able to treat a wide range of diseases such as fever, cold, or even cancer. People usually take it by mixing small drops with camel milk or take it directly. The project aims to study the effects of camel urine in inhibiting the growth potential and metastatic ability of 4T1 cancer cell line in vitro and in vivo. Based on the MTT result, the cytotoxicity of camel urine against 4T1 cell was established, and it was dose-dependent. Additionally, the antimetastatic potential of camel urine was tested by running several assays such as scratch assay, migration and invasion assay, and mouse aortic ring assay with promising results in the ability of camel urine to inhibit metastatic process of the 4T1 cells. In order to fully establish camel urine’s potential, an in vivo study was carried out by treating mice inoculated with 4T1 cells with 2 different doses of camel urine. By the end of the treatment period, the tumor in both treated groups had reduced in size as compared to the control group. Additional assays such as the TUNEL assay, immunophenotyping, cytokine level detection assay, clonogenic assay, and proteome profiler demonstrated the capability of camel urine to reduce and inhibit the metastatic potential of 4T1 cells in vivo. To sum up, further study of anticancer properties of camel urine is justified, as evidenced through the in vitro and in vivo studies carried out. Better results were obtained at higher concentration of camel urine used in vivo. Apart from that, this project has laid out the mechanisms employed by the substance to inhibit the growth and the metastatic process of the 4T1 cell

Packages of Care for Epilepsy in Low- and Middle-Income Countries

In the second in a series of six articles on packages of care for mental health disorders in low- and middle-income countries, Caroline Mbuba and Charles Newton discuss treatment for epilepsy

Bi-allelic <em>ACBD6</em> variants lead to a neurodevelopmental syndrome with progressive and complex movement disorders

\ua9 The Author(s) 2023. Published by Oxford University Press on behalf of the Guarantors of Brain. The acyl-CoA-binding domain-containing protein 6 (ACBD6) is ubiquitously expressed, plays a role in the acylation of lipids and proteins and regulates the N-myristoylation of proteins via N-myristoyltransferase enzymes (NMTs). However, its precise function in cells is still unclear, as is the consequence of ACBD6 defects on human pathophysiology. Using exome sequencing and extensive international data sharing efforts, we identified 45 affected individuals from 28 unrelated families (consanguinity 93%) with bi-allelic pathogenic, predominantly loss-of-function (18/20) variants in ACBD6. We generated zebrafish and Xenopus tropicalis acbd6 knockouts by CRISPR/Cas9 and characterized the role of ACBD6 on protein N-myristoylation with myristic acid alkyne (YnMyr) chemical proteomics in the model organisms and human cells, with the latter also being subjected further to ACBD6 peroxisomal localization studies. The affected individuals (23 males and 22 females), aged 1-50 years, typically present with a complex and progressive disease involving moderate-to-severe global developmental delay/intellectual disability (100%) with significant expressive language impairment (98%), movement disorders (97%), facial dysmorphism (95%) and mild cerebellar ataxia (85%) associated with gait impairment (94%), limb spasticity/hypertonia (76%), oculomotor (71%) and behavioural abnormalities (65%), overweight (59%), microcephaly (39%) and epilepsy (33%). The most conspicuous and common movement disorder was dystonia (94%), frequently leading to early-onset progressive postural deformities (97%), limb dystonia (55%) and cervical dystonia (31%). A jerky tremor in the upper limbs (63%), a mild head tremor (59%), parkinsonism/hypokinesia developing with advancing age (32%) and simple motor and vocal tics were among other frequent movement disorders. Midline brain malformations including corpus callosum abnormalities (70%), hypoplasia/agenesis of the anterior commissure (66%), short midbrain and small inferior cerebellar vermis (38% each) as well as hypertrophy of the clava (24%) were common neuroimaging findings. Acbd6-deficient zebrafish and Xenopus models effectively recapitulated many clinical phenotypes reported in patients including movement disorders, progressive neuromotor impairment, seizures, microcephaly, craniofacial dysmorphism and midbrain defects accompanied by developmental delay with increased mortality over time. Unlike ACBD5, ACBD6 did not show a peroxisomal localization and ACBD6-deficiency was not associated with altered peroxisomal parameters in patient fibroblasts. Significant differences in YnMyr-labelling were observed for 68 co- and 18 post-translationally N-myristoylated proteins in patient-derived fibroblasts. N-myristoylation was similarly affected in acbd6-deficient zebrafish and X. tropicalis models, including Fus, Marcks and Chchd-related proteins implicated in neurological diseases. The present study provides evidence that bi-allelic pathogenic variants in ACBD6 lead to a distinct neurodevelopmental syndrome accompanied by complex and progressive cognitive and movement disorders

Proteins with Complex Architecture as Potential Targets for Drug Design: A Case Study of Mycobacterium tuberculosis

Lengthy co-evolution of Homo sapiens and Mycobacterium tuberculosis, the main causative agent of tuberculosis, resulted in a dramatically successful pathogen species that presents considerable challenge for modern medicine. The continuous and ever increasing appearance of multi-drug resistant mycobacteria necessitates the identification of novel drug targets and drugs with new mechanisms of action. However, further insights are needed to establish automated protocols for target selection based on the available complete genome sequences. In the present study, we perform complete proteome level comparisons between M. tuberculosis, mycobacteria, other prokaryotes and available eukaryotes based on protein domains, local sequence similarities and protein disorder. We show that the enrichment of certain domains in the genome can indicate an important function specific to M. tuberculosis. We identified two families, termed pkn and PE/PPE that stand out in this respect. The common property of these two protein families is a complex domain organization that combines species-specific regions, commonly occurring domains and disordered segments. Besides highlighting promising novel drug target candidates in M. tuberculosis, the presented analysis can also be viewed as a general protocol to identify proteins involved in species-specific functions in a given organism. We conclude that target selection protocols should be extended to include proteins with complex domain architectures instead of focusing on sequentially unique and essential proteins only