Drug Sensitivity Patterns Research Articles

APOBEC family reshapes the immune microenvironment and therapy sensitivity in clear cell renal cell carcinoma

Emerging evidence suggests that the APOBEC family is implicated in multiple cancers and might be utilized as a new target for cancer detection and treatment. However, the dysregulation and clinical implication of the APOBEC family in clear cell renal cell cancer (ccRCC) remain elusive. TCGA multiomics data facilitated a comprehensive exploration of the APOBEC family across cancers, including ccRCC. Remodeling analysis classified ccRCC patients into two distinct subgroups: APOBEC family pattern cancer subtype 1 (APCS1) and subtype 2 (APCS2). The study investigated differences in clinical parameters, tumor immune microenvironment, therapeutic responsiveness, and genomic mutation landscapes between these subtypes. An APOBEC family-related risk model was developed and validated for predicting ccRCC patient prognosis, demonstrating good sensitivity and specificity. Finally, the overview of APOBEC3B function was investigated in multiple cancers and verified in clinical samples. APCS1 and APCS2 demonstrated considerably distinct clinical features and biological processes in ccRCC. APCS1, an aggressive subtype, has advanced clinical stage and a poor prognosis. APCS1 exhibited an oncogenic and metabolically active phenotype. APCS1 also exhibited a greater tumor mutation load and immunocompromised condition, resulting in immunological dysfunction and immune checkpoint treatment resistance. The genomic copy number variation of APCS1, including arm gain and loss, was much more than that of APCS2, which may help explain the tired immune system. Furthermore, the two subtypes have distinct drug sensitivity patterns in clinical specimens and matching cell lines. Finally, we developed a predictive risk model based on subtype biomarkers that performed well for ccRCC patients and validated the clinical impact of APOBEC3B. Aberrant APOBEC family expression patterns might modify the tumor immune microenvironment by increasing the genome mutation frequency, thus inducing an immune-exhausted phenotype. APOBEC family-based molecular subtypes could strengthen the understanding of ccRCC characterization and guide clinical treatment. Targeting APOBEC3B may be regarded as a new therapeutic target for ccRCC.

Bullet-related bacterial wound infections among injured personnel at emergency site hospitals in Bahir Dar: prevalence, antimicrobial susceptibility and associated factors.

Bullet-related bacterial wound infection can be caused by high-velocity bullets and shrapnel injuries. In Ethiopia, significant injuries were reported that may cause severe wound infections, persistent systemic infections and may lead to amputation and mortality. The magnitude, antimicrobial susceptibility profiles, and factors associated with bacterial wound infections among patients with bullet-related injuries are not yet studied particularly at health facilities in Bahir Dar, Northwest Ethiopia. Therefore, this study was aimed to determine the prevalence, bacterial profiles, antimicrobial susceptibility profiles, and factors associated with bacterial infections among patients with bullet-related injuries at referral health facilities in Bahir Dar, Northwest Ethiopia. A Hospital-based cross-sectional study was conducted among patients with bullet-related injuries at three referral health facilities in Bahir Dar from May 25 to July 27, 2022. A total of 384 patients with bullet-related injuries were included in the study. Sociodemographic and clinical data were collected using a structured questionnaire. Wound swabs were collected aseptically and cultured on Blood and MacConkey agar following bacteriological standards. Biochemical tests were performed to differentiate bacteria for positive cultivation and antimicrobial susceptibility profiles of the isolates were done on Muller Hinton agar using the Kirby-Bauer disk diffusion technique according to the 2021 Clinical Laboratory Standard Institute (CLSI) guideline. The data were entered using Epi-Info version 7.3 and analyzed using SPSS version 25. Descriptive data were presented using frequency, percentages, figures, and charts. Logistic regression was carried out to identify factors associated with bacterial wound infections. P-value < 0.05 was considered statistically significant. The prevalence of bullet-related bacterial wound infection among three referral hospitals in Bahir Dar city was 54.7%. The most commonly isolated Gram-negative organism was Klebsiella spps 49 (23.3%) while among Gram-positive organism, Staphylococcus aureus 58 (27.6%) and coagulase-negative staphylococci (CONS) 18 (8.6%). Contamination, hospitalization and smoking habit were significantly associated with the presence of bullet-related bacterial wound infections. Over 97% multidrug resistant (MDR) bacterial isolates were identified and of theses, E. coli, Proteus species, Citrobactor, and Staphylococcus aureus were highly drug resistant. Increased prevalence of bullet-related bacterial wound infection was noticed in this study. S. aureus followed by Klebsiella species were most commonly isolated bacteria. High frequency of resistance to Ampicillin, Oxacillin, Cefepime, Ceftriaxone, Ceftazidime, Vancomycin, and Norfloxacin was observed. Therefore, proper handling of bullet injuries, prompt investigation of bacterial infections, monitoring of drug sensitivity patterns and antibiotic usage are critical.

Microbiology and drug sensitivity of Surgical Site Infections in Pakistan: A Systematic Review

Abstract Various bacterial pathogens that are known to cause Surgical Site Infections (SSI) are becoming increasingly resistant to many antibiotics. The aim of this systematic review was to see various pathogens responsible to cause SSI in Pakistan and to see their drug sensitivity patterns according to culture and sensitivity reports during last five years. This systematic review was conducted according to the preferred reporting items for systematic reviews and met analysis (PRISMA) guidelines. Extensive search of medical databases Pubmed, Google Scholar and Pakmedinet was done. Four studies were selected from various cities of Pakistan. Most common bacterial pathogen isolated was staph aureus 49%, followed by Ecoli 18%, Enterococcus 8%, and Pseudomonas 7.4%. Methicillin resistant staph aureus (MRSA) was isolated as 5% and 18 % in two studies. Drug sensitivity pattern showed Vancomycin, Imipenem, Meropenem, Linezolid, Cefoperazone + Sulbactam to be the most sensitive drugs against all type of pathogens. Drugs found effective against staph aureus were Vancomycin, Linezolid, Meropenem and Fusidic acid. Drugs found sensitive against E coli were Cefoperazone Sulbactam combination, Linezolid and Meropenem. Ampicillin and Amoxicillin were found highly resistant against all pathogens. We concluded from our systematic review that most common bacterial pathogen responsible to cause SSI in Pakistan is staph aureus. Empirically Linezolid is found effective against all pathogens. Imipenem, Meropenem, sulbactam plus Cefoperazone combination, vancomycin, tazocin are effective drugs. Ampicillin, amoxicillin and ciprofloxacin are highly resistant in Pakistan. Key words: Surgical Site Infections (SSI), Staph aureus, E Coli, Culture and Sensitivity

Study of utilization of antimicrobial drugs and its resistance pattern in patients with septicemia at a tertiary care hospital

Background: Many types of bacteria can produce septicemia. The most common infections that lead to septicemia are urinary tract infections, pneumonia, nephritis and abdominal infections. Methods: The study aims to assess the pattern of antimicrobial drugs used in septicemia and to assess the etiological organisms and their drug sensitivity and resistance pattern. Study participants admitted to tertiary health care centre and who are having septicemia were included in this prospective observational study. Blood culture, bronchial secretions were subjected to microbiological analysis. The etiological organisms, their drug sensitivity and resistance pattern and the outcome of drug therapy were recorded. The clinical course of the study participants was monitored till cure either the resolution of pneumonia. Qualitative data were analyzed using the chi-square test or Fischer's exact test and quantitative data using the independent t test. Results: Mean duration of hospital stay in all patients 13.22±0.45 days and in resistance cases it was found 19.22±0.45 days. Mean duration of ICU stay in all patients was 4.34±0.45 days and in resistant cases mean duration of ICU stay was 8.18±0.45 days. Clinical outcome in overall admitted patients recovered was 97 patients (80.83%) death was 16 patients (13.33%). Clinical outcome in resistant patients recovered 16 patients (13.33%) and death 26 patients (21.66%). Conclusions: The organisms had a varied sensitivity and resistance pattern. The clinical outcome was multifactorial.

Evaluation of the Types and Drug Sensitivity Patterns of Bacterial Isolates from Patients with Eye Discharge at Abia State University Teaching Hospital, Aba, Nigeria

This study aimed to evaluate the types and drug sensitivity patterns of bacterial isolates from patients with eye discharge at the Abia State University Teaching Hospital (ABSUTH), Aba, Nigeria. A total of 100 samples were collected, with the most prevalent bacterial isolates being Staphylococcus aureus (26%) and Coagulase-negative Staphylococci (21%), while no bacterial growth was reported in 42% of samples. In terms of patient demographics, a significant proportion of the patients were females (64%) aged between 61-70 years. In relation to contact lens use, a high percentage (97%) of the bacterial isolates were detected in patients who did not use contact lenses. An occupational distribution analysis revealed that bacterial isolates were more frequent in farmers and traders compared to students and retirees. Antimicrobial sensitivity testing of the bacterial isolates showed varying degrees of resistance, with significant resistance observed against Erythromycin, Ciprofloxacin, and Perfloxacin. In contrast, higher sensitivity was noted towards Chloramphenicol and Gentamycin. The findings underscore the importance of regular microbiological evaluation and sensitivity testing in patients presenting with eye discharges to guide effective antimicrobial therapy. These findings could guide the selection of empiric antibiotics for treating ocular infections in this region and suggest the need for routine microbiological surveillance to monitor and guide antibiotic prescription to mitigate against antibiotic resistance.

Comprehensive somatic mutational analysis in glioblastoma: Implications for precision medicine approaches.

Glioblastoma multiforme (GBM), a malignant neoplasm originating from glial cells, remains challenging to treat despite the current standard treatment approach that involves maximal safe surgical resection, radiotherapy, and adjuvant temozolomide chemotherapy. This underscores the critical need to identify new molecular targets for improved therapeutic interventions. The current study aimed to explore the somatic mutations and potential therapeutic targets in GBM using somatic mutational information from four distinct GBM datasets including CGGA, TCGA, CPTAC and MAYO-PDX. The analysis included the evaluation of whole exome sequencing (WES) of GBM datasets, tumor mutation burden assessment, survival analysis, drug sensitivity prediction, and examination of domain-specific amino acid changes. The results identified the top ten commonly altered genes in the aforementioned GBM datasets and patients with mutations in OBSCN and AHNAK2 alone or in combination had a more favorable overall survival (OS). Also, the study identified potential drug sensitivity patterns in GBM patients with mutations in OBSCN and AHNAK2, and evaluated the impact of amino acid changes in specific protein domains on the survival of GBM patients. These findings provide important insights into the genetic alterations and somatic interactions in GBM, which could have implications for the development of new therapeutic strategies for this aggressive malignancy.

Current Status of Drug-resistant Patterns of Gram-positive Clinical Isolates Collected from Renowned Diagnostic Centers of Dhaka, Bangladesh

Background: Antimicrobial resistance is a serious concern and life threat in public health globally. Gram-positive clinical isolates are one of the leading etiological agents showing drug resistance traits. This study aimed to detect the antimicrobial resistance pattern of clinically isolated Gram-positive bacterial isolates collected from various renowned diagnostic centers of Dhaka city, Bangladesh. Methods: A laboratory-based cross-sectional study was conducted at the Microbiological Laboratory of the Department of Microbiology, Stamford University Bangladesh. A total of 104 clinical specimens (wound swab, pus, blood, urine, ear discharge, nasal swab, sputum, nipple discharge, breast milk, catheter tip, umbilical swab, and throat swab) from different age groups were collected from some renowned diagnostic centers of Dhaka city for analyzing the cultural and drug sensitivity patterns from January 2023 to April 2023. According to the study plan, only the Gram-positive bacteria were collected and transported to the laboratory of the Department of Microbiology, Stamford University Bangladesh following WHO guidelines, and the antimicrobial susceptibility tests were carried out by Kirby–Bauer disk diffusion method and the results were interpreted as per CLSI guidelines, 2022. Results: This study highlights the presence of Staphylococcus aureus (65.4%), Streptococcus spp. (23.1%), and Enterococcus spp. (11.5%) as causative agents of Gram-positive bacterial infections from overall 104 clinical specimens. Most of our isolates were found multidrug-resistant. For S. aureus isolates, cefixime (100%); for Streptococcus spp., doxycycline (95.83%); and for Enterococcus spp., amoxicillin (100%), cefixime (100%), colistin (100%), and cotrimoxazole (100%) were found to be the most ineffective drug when compared to other antibiotics. Conclusion: Our study portrayed the remarkable antibiotic resistance feature of Gram-positive bacterial isolates which is a matter of public health concern. It is necessary to carry out a continuous assessment of antibiotic sensitivity patterns of resistant Gram-positive bacteria; otherwise, it will become an uncontrollable problem not only in Bangladesh but also worldwide in the near future.

Microbiological Evaluation of Paediatric Chronic Haematogenous Osteomyelitis in a Tertiary Care Hospital in Northern India and its Association with Radiological Appearance: A Retrospective Study

Introduction: Chronic osteomyelitis in the paediatric age group is a frequent complication of improperly treated acute haematogenous osteomyelitis, leading to devastating complications such as pathological fractures and deformities in affected children. There is a growing need to comprehend the disease process and develop improved treatment strategies. Aim: To evaluate bacterial culture and antibiotic sensitivity patterns in children and adolescents with chronic haematogenous osteomyelitis in North India and to explore associations between radiological patterns and antibiotic sensitivity. Materials and Methods: The present retrospective study was conducted in the Department of Orthopaedics of tertiary care centre, Himalayan Institute of Medical Sciences, Dehradun, Uttarakhand, North India, from July 2018 to June 2020. Total 100 children and young adolescents with chronic osteomyelitis who reported to OPD of hospital with pus discharge from an extremity persisting for more than six weeks, along with radiological features indicative of chronic osteomyelitis. Parameters such as site of infection, illness duration, blood parameters, bacteriological culture reports, etc., were examined. Radiological evaluation was performed using the Beit CURE (BC) classification. Data were analysed and represented in the form of frequencies and percentages. The Chi-square test was used to compare proportions, with significance set at p-value&lt;0.05. Results: The median age of the study population was 13 years, there were 32 (32%) females and 68 (68%) males. The majority of cases (70%) belonged to the B1-B3 group. Staphylococcus aureus was responsible for 86 (86%) cases, followed by Pseudomonas aeruginosa in 7 (7%) cases. Out of the 86 Staphylococcus aureus (S. aureus) isolates, 58 (67.44%) were Methicillin-resistant Staphylococcus aureus (MRSA). Multidrug resistance was observed among gram-negative species, as well. Pseudomonas aeruginosa showed resistance to fluoroquinolones in 4 (57%) cases, aminoglycosides in 2 (28%) cases, and carbapenem in 1 (14%) case, which is considered highly effective in treating serious infections caused by multidrug resistant Gram-negative species. No significant association was found between drug sensitivity patterns and radiological features in the present study. Conclusion: Staphylococcus aureus remains the most predominant organism isolated from deep tissue cultures. Among S. aureus isolates, MRSA was the most frequently identified. Identifying the causative organism may be challenging in some cases. Among cases where the offending microbe was identified, drug resistance was widespread among both Gram-negative and Gram-positive specimens. No significant relationship was found between the radiological appearance of infected bone and the pattern of antibiotic resistance.

Increased sensitivity of malaria parasites to common antimalaria drugs after the introduction of artemether-lumefantrine: Implication of policy change and implementation of more effective drugs in fight against malaria.

Single nucleotide polymorphisms (SNPs) in the Plasmodium falciparum multi-drug resistance protein 1 (Pfmrp1) gene have previously been reported to confer resistance to Artemisinin-based Combination Therapies (ACTs) in Southeast Asia. A total of 300 samples collected from six sites between 2008 and 2019 under an ongoing malaria drug sensitivity patterns in Kenya study were evaluated for the presence of SNPs at Pfmrp1 gene codons: H191Y, S437A, I876V, and F1390I using the Agena MassARRAY® platform. Each isolate was further tested against artemisinin (ART), lumefantrine (LU), amodiaquine (AQ), mefloquine (MQ), quinine (QN), and chloroquine (CQ) using malaria the SYBR Green I-based method to determine their in vitro drug sensitivity. Of the samples genotyped, polymorphism at Pfmrp1 codon I876V was the most frequent, with 59.3% (163/275) mutants, followed by F1390I, 7.2% (20/278), H191Y, 4.0% (6/151), and S437A, 3.3% (9/274). A significant decrease in median 50% inhibition concentrations (IC50s) and interquartile range (IQR) was noted; AQ from 2.996 ng/ml [IQR = 2.604-4.747, n = 51] in 2008 to 1.495 ng/ml [IQR = 0.7134-3.318, n = 40] (P<0.001) in 2019, QN from 59.64 ng/ml [IQR = 29.88-80.89, n = 51] in 2008 to 18.10 ng/ml [IQR = 11.81-26.92, n = 42] (P<0.001) in 2019, CQ from 35.19 ng/ml [IQR = 16.99-71.20, n = 30] in 2008 to 6.699 ng/ml [IQR = 4.976-9.875, n = 37] (P<0.001) in 2019, and ART from 2.680 ng/ml [IQR = 1.608-4.857, n = 57] in 2008 to 2.105 ng/ml [IQR = 1.266-3.267, n = 47] (P = 0.0012) in 2019, implying increasing parasite sensitivity to the drugs over time. However, no significant variations were observed in LU (P = 0.2692) and MQ (P = 0.0939) respectively, suggesting stable parasite responses over time. There was no statistical significance between the mutation at 876 and parasite sensitivity to selected antimalarials tested, suggesting stable sensitivity for the parasites with 876V mutations. These findings show that Kenyan parasite strains are still sensitive to AQ, QN, CQ, ART, LU, and MQ. Despite the presence of Pfmrp1 mutations in parasites among the population.

Erysipelothrix rhusiopathiae: An Emerging Organism Causing Pleural Effusion in an Immunocompetent Child

Aims: Erysipelothrix rhusiopathiae is a Gram-positive bacillus attributed to zoonotic infections primarily. In humans they usually cause localized cutaneous infections namely erysipeloid, occasionally becomes diffuse and may lead to septicemia and endocarditis in opportunistic immunocompromised individuals. The growing and evolving disease spectrum of this organism in human beings is a potential future threat. This case report addresses a common disease manifestation with this unusual organism which is needs to be highlighted.&#x0D; Presentation of Case: Herein we report a case of 1year 9month old immunocompetent male child belonging to a shepherd family who presented with long standing left sided pleural effusion. Pleural fluid culture yielded Erysipelothrix rhusiopathiae. Despite a culture sensitive antibiotic therapy a sequelae of residual effusion with subpleural collapse and fibrous septations developed, hence left lateral thoracotomy along with decortication surgery was performed.&#x0D; Discussion: This case report highlights the drug sensitivity and resistance pattern in detail and also points out the prognosis and possible sequelae of the disease which in turn can increase treatment awareness regarding the organism.&#x0D; Conclusion: Erysipelothrix rhusiopathiae is an unusual organism which was found in our patient causing pleural effusion is a very unlikely coalition and needs a separate mention as this emerging microbe can be a cause of concern due to its novelty and scarce medical evidence.

Functional and Molecular Heterogeneity in Glioma Stem Cells Derived from Multiregional Sampling.

Glioblastoma (GBM) is an aggressive and highly heterogeneous primary brain tumor. Glioma stem cells represent a subpopulation of tumor cells with stem cell traits that are presumed to be the cause of tumor relapse. There exists complex tumor heterogeneity in drug sensitivity patterns between glioma stem cell (GSC) cultures derived from different patients. Here, we describe that heterogeneity also exists between GSC cultures derived from multiple biopsies within a single tumor. From biopsies harvested within spatially distinct regions representing the entire tumor mass, we established seven GSC cultures and compared their stem cell properties, mutations, gene expression profiles, and drug sensitivity patterns against 115 different anticancer drugs. The results were compared to 14 GSC cultures derived from other patients. Between the multiregional-derived GSC cultures, we observed only minor differences in their phenotype, proliferative capacity, and global gene expression. Further, they displayed intratumoral heterogeneity in mutational profiles and sensitivity patterns to anticancer drugs. This heterogeneity, however, did not exceed the extensive heterogeneity found between GSC cultures derived from other GBM patients. Our results suggest that the use of GSC cultures from one single focal biopsy may underestimate the overall complexity of the GSC population and display the importance of including GSC cultures reflecting the entire tumor mass in drug screening strategies.

Burden of enteric fever and antibiotic sensitivity in Nepalese Children Prior to Typhoid Vaccine in National Immunization Program.

Enteric fever is a major public health problem in developing and under developed countries. Case fatality rate without treatment is 10-30% and with appropriate treatment is only 1-4%. Gold standard for diagnosis is isolation of Salmonella enterica from blood or bone marrow. Antibiotics resistance is skyrocketing with emergence of multidrug resistance S. typhi and extensively drug resistant S. typhi. The blood culture done in Kanti children hospital in last six years were taken from the data base and the culture positive cases were taken from which the salmonella species positive cases along with the drug sensitivity pattern were used in our study. The culture positivity rate was 2.8% and 7.6% (n=136) among the culture positive cases were Salmonella species. Salmonella typhi (121; 88.9%) was the most frequently isolated species, followed by Salmonella paratyphi A (13; 9.5%) and Salmonella paratyphi B (2;1.4%). Children with age 5-10 years was the most affected age group for infection with Salmonella, 50.0% (n=68). Nalidixic acid is resistant in 89.9% Salmonella typhi; followed by ciprofloxacin (31.8%), ofloxacin (18.2%), ampicillin (9.6%), azithromycin (8.4%), chloramphenicol (8.2%), cotrimoxazole (5.4%), cefixime (4%), ceftriaxone (2.5%) and cefotaxime (0.0%). Cefixime, ceftriaxone, cefotaxime are 100% sensitive to Salmonella paratyphi, followed by cotrimoxazole (92.9%), ofloxacin (81.8%), chloramphenicol (75%), azithromycin (66.7%), ampicillin (60%), ciprofloxacin (50%) and Nalidixic acid (23.1%). Salmonella species culture isolatation are declining every year. Fluoroquinolones have more resistance than first line drugs of typhoid, azithromycin resistance is rising but 3rd generation cephalosporins are sensitive to Salmonella species.

The Tetraspanin CD9 Shapes Glucocorticoid Sensitivity in Pediatric B-Precursor Acute Lymphoblastic Leukemia

Background: De novo or acquired resistance to glucocorticoids (GCs), the common therapeutic regimens for remission-induction in pediatric B-precursor acute lymphoblastic leukemia, confronts treatment success and is a strong predictor for disease relapse. Discovery of predictive markers and underlying mechanisms of GC resistance could lead to new treatment and prognostic modalities. Aims: This study aims: (1) to demonstrate the association between CD9 expression and GC sensitivity in childhood BCP-ALL; (2) to dissect the underlying mechanisms driving GC resistance; and (3) to discover the possible therapeutic modalities for restoring GC susceptibility. Methods: Ex vivo drug sensitivity profiling to prednisone (Pred) and dexamethasone (Dex) was performed on 6 BCP-ALL cell lines and 18 patient samples with differential CD9 expression. Association of CD9 expression with patient responses to Pred prophase was analyzed in a pediatric BCP-ALL cohort (n=182). Functional impact of CD9 on GC susceptibility was dissected by gain- and loss-of-function experiments in BCP-ALL cells. Mechanistic basis of CD9-driven GC response was investigated through genomics, transcriptomics, and proteomics. Reversal of GC resistance by pathway-targeted agents were explored on BCP-ALL cells and xenograft models. Results: Distinctive differences in the drug sensitivity pattern were found between CD9- and CD9+ BCP-ALL cell lines, with the former showing resistance to GCs but not other chemotherapeutic agents. Concurrently, CD9- primary lymphoblasts exhibited resistance to Dex (IC50s: 2,761 vs. 31 nM) and Pred (IC50s: 8,185 vs. 268 nM) compared with CD9+ samples (positivity defined by the presence of ≥ 20% CD9+ blasts). More Pred poor responders (defined by the presence of ≥ 1×10 9/L circulating lymphoblasts on day 8) were found in CD9- than CD9+ patients (19.4% vs. 6.2%, P=0.018). Subgroup analyses revealed more poor responders with CD9- phenotype were enriched in patients with older age (60% vs. 3.7%, P=0.008), male gender (21.7% vs. 6.7%, P=0.045), higher white blood cell count (66.7% vs. 15.4%, P=0.004), and in those with BCR-ABL1 translocation (100% vs. 12.5%, P=0.024), or not otherwise cytogenetically specified BCP-ALL (30.8% vs. 5.8%, P=0.019). Univariate and multivariate analyses further confirmed CD9 negativity as an independent predictor for GC resistance (OR=5.1, P=0.009). Overexpression of CD9 in CD9- cells (SEM, KOPN-8) by lentiviral transduction significantly enhanced GC sensitivity by 2.8-62.3-fold ( P&amp;lt;0.024), whereas CD9 knockout in CD9+ cells (697) using CRISPR-Cas9 decreased GC susceptibility ( P&amp;lt;0.003). No significant differences in mRNA or protein expression of the GC receptor NR3C1 were identified between CD9+ and CD9- cells, as well as its isoforms or mutants. The cytoplasm-to-nucleus translocation of GC receptor was consistent among cell lines regardless of CD9 expression status, but CD9 was physically interacted with NR3C1. Upon GC stimulation, the presence of CD9 increased the number of DEGs (110 vs. 82 DEGs) and magnitude of GC-responsive genes exclusively detected in the background of CD9 (1.3-18.6-fold; e.g. AKAP13, STAT5A, EZR). CD9- cells showed constitutive activation of the MAPK pathway and could be preferentially restored for GC susceptibility by the MEK1/2 inhibitor trametinib in vitro. BCP-ALL xenograft modeling revealed that, compared to single agent Dex treatment, combination with trametinib significantly reduced the leukemic load in animal transplanted with CD9- SEM but not CD9+ BV-173 cell lines. Ex vivo drug testing further showed that the resistance to GCs in some CD9+ cases could be partially reversed by combination of trametinib and the JAK inhibitor ruxolitinib due to the activation of STAT5. Conclusions: This study provides a definitive linkage of CD9 with GC sensitivity in childhood BCP-ALL mediated through a NR3C1-independent mechanism. CD9- cases could be prone to trametinib combination therapy, whereas CD9+ cases might need a third drug such as ruxolitinib to counteract GC resistance.

Delayed Intensification Including Venetoclax and Bortezomib Prolongs Survival in Very High Risk Acute Lymphoblastic Leukaemia

Modern multi-drug risk-stratified response-adapted chemotherapy protocols lead to &amp;gt;85% cure rates in childhood acute lymphoblastic leukaemia (ALL). Between 2013-2022, 707 children (0-18 years) with acute lymphoblastic leukaemia (ALL) treated systematically on the ICiCLe-ALL-14 risk-adapted protocol achieved a 3-year event-free survival (EFS) of 73% (95% confidence interval, CI 69-77). A very high risk (VHR) subgroup (N = 107) was identified with poor outcome (3-year EFS, 38% [95% CI 25-51]). This included patients with induction failure, persistent minimal residual disease (MRD) and/or select high risk cytogenetic subtypes ( BCR::ABL1 other ABL-class fusion ALL, TCF3::HLF, low hypodiploidy, IKZF1plus with high end-induction MRD, KMT2A-rearranged infant ALL). Co-culture based high-throughput drug response profiling (DRP) was performed to identify alternate chemotherapeutic drugs for these VHR patients. Primary ALL blasts extracted by density centrifugation, were seeded on immortalized human bone marrow stromal cells and treated with a 14-drug panel: prednisolone; vincristine (VCR), daunorubicin; asparaginase (ASNase), bortezomib (BZM), venetoclax (VEN), panobinostat (PNB), mitoxantrone, cyclophosphamide, cytarabine, selinexor, dasatinib, 6-thioguanine and idarubicin. Seventy-four patients (25 VHR, 37 high-risk, 7 intermediate-risk, and 5 standard-risk,) were analysed. Venetoclax, BZM and PNB were identified as active agents (IC 50 &amp;lt;100nM for VEN and PNB, &amp;lt;10nM for BZM) in 71%, 42% and 75% of VHR samples, respectively. Based on drug-sensitivity patterns of VHR ALL, a pilot study was initiated to prospectively evaluate the addition of VEN and BZM in the frontline delayed intensification (DI) phase of VHR ALL patients (N=24) after informed consent (Figure 1). Response was assessed before and after VHR-DI in 46% (11/24) patients. Good response, i.e. decrease in MRD ≥ 1 log after VHR-DI, was noted in 82% (9/11) patients. Landmark analysis comparing survival of patients (from start of DI) treated on standard DI versus VHR-DI showed significant improvement in 2-year EFS from 54% (95% CI 40-66) to 76% (95% CI 51-89) ( p=0.037, log-rank test). Overall, VHR-DI was tolerated well. No significant difference was noted in grade 4 and 5 toxicities (Common Terminology Criteria for Adverse Events v4.0) between standard DI and VHR-DI ( p= 0.053 &amp; p=0.988 respectively, chi-square test). Fever-neutropenia and sepsis were the most frequent toxicities with VHR-DI. To identify potential mechanisms of resistance in poor-responders, transcriptome analysis was performed on diagnostic blast cells from VHR patients with good (N=5) or poor (N=5) response to VHR-DI and compared with highly sensitive SR patients (N=5). Upregulation of oxidative phosphorylation and ubiquitin-mediated proteolysis were found in good responders while non-responders showed upregulation of amino acid metabolic and RNA splicing pathways. In conclusion, drug response profiling identifies alternative combination therapies for VHR ALL. Our pilot study provides proof-of-concept of effectiveness and tolerability of inclusion of VEN and BZM in the post-induction treatment of patients with poor-response disease.

Identification of the shared gene signature and biological mechanism between type 2 diabetes and colorectal cancer.

Background: The correlation of type 2 diabetes mellitus (T2DM) with colorectal cancer (CRC) has garnered considerable attention in the scientific community. Despite this, the molecular mechanisms underlying the interaction between these two diseases are yet to be elucidated. Hence, the present investigation aims to explore the shared gene signatures, immune profiles, and drug sensitivity patterns that exist between CRC and T2DM. Methods: RNA sequences and characteristics of patients with CRC and T2DM were retrieved from The Cancer Genome Atlas and Gene Expression Omnibus databases. These were investigated using weighted gene co-expression network analysis (WGCNA) to determine the co-expression networks linked to the conditions. Genes shared between CRC and T2DM were analyzed by univariate regression, followed by risk prognosis assessment using the LASSO regression model. Various parameters were assessed through different software such as the ESTIMATE, CIBERSORT, AND SSGSEA utilized for tumor immune infiltration assessment in the high- and low-risk groups. Additionally, pRRophetic was utilized to assess the sensitivity to chemotherapeutic agents in both groups. This was followed by diagnostic modeling using logistic modeling and clinical prediction modeling using the nomogram. Results: WGCNA recognized four and five modules that displayed a high correlation with T2DM and CRC, respectively. In total, 868 genes were shared between CRC and T2DM, with 14 key shared genes being identified in the follow-up analysis. The overall survival (OS) of patients in the low-risk group was better than that of patients in the high-risk group. In contrast, the high-risk group exhibited higher expression levels of immune checkpoints The Cox regression analyses established that the risk-score model possessed independent prognostic value in predicting OS. To facilitate the prediction of OS and cause-specific survival, the nomogram was established utilizing the Cox regression model. Conclusion: The T2DM + CRC risk-score model enabled independent prediction of OS in individuals with CRC. Moreover, these findings revealed novel genes that hold promise as therapeutic targets or biomarkers in clinical settings.

Comprehensive analysis of ferroptosis-related genes in immune infiltration and prognosis in multiple myeloma.

Background: One particular type of cellular death that is known as ferroptosis is caused by the excessive lipid peroxidation. It is a regulated form of cell death that can affect the response of the tumor cells. Currently, it is not known if the presence of this condition can affect the prognosis of patients with multiple myeloma (MM). Methods: In this study, we studied the expression differences and prognostic value of ferroptosis-related genes (FRGs) in MM, and established a ferroptosis risk scoring model. In order to improve the prediction accuracy and clinical applicability, a nomogram was also established. Through gene enrichment analysis, pathways closely related to high-risk groups were identified. We then explored the differences in risk stratification in drug sensitivity and immune patterns, and evaluated their value in prognostic prediction and treatment response. Lastly, we gathered MM cell lines and samples from patients to confirm the expression of marker FRGs using quantitative real-time PCR (qRT-PCR). Results: The ability to predict the survival of MM patients is a challenging issue. Through the use of a risk model derived from ferroptosis, we were able to develop a more accurate prediction of the disease's prognosis. They were then validated by a statistical analysis, which showed that the model is an independent factor in the prognosis of MM. Patients of high ferroptosis risk scores had a much worse chance of survival than those in the low-risk groups. The calibration and power of the nomogram were also strong. We noted that the link between the ferroptosis risk score and the clinical treatment was suggested by the FRG's significant correlation with the immune checkpoint genes and the medication sensitivity. We validated the predictive model using qRT-PCR. Conclusion: We demonstrated the association between FRGs and MM, and developed a new risk model for prognosis in MM patients. Our study sheds light on the potential clinical relevance of ferroptosis in MM and highlights its potential as a therapeutic target for patients with this disease.

A study of clinico-mycological profile and treatment of primary otomycosis

Background: Otomycosis is a superficial mycotic infection of the external auditory canal. Clinical presentation, along with otoscopic findings, is sufficient to make the diagnosis in most of the cases. Treatment includes aural cleaning followed by the instillation of anti-fungal ear drops. In the recent years, there has been an increasing trend of antifungal resistance for the commonly used antifungals such as fluconazole and clotrimazole. Drug sensitivity patterns have also shown variability from one region to another. Aims and Objectives: To evaluate the clinico-mycological profile along with antifungal susceptibility pattern and treatment outcome of primary otomycosis. Materials and Methods: A total of 230 clinically diagnosed patients of primary otomycosis were included in this prospective study. Microbiological causes were evaluated by KOH wet mount and culture of samples collected from external auditory meatus using sterile swabs. Fungal culture was done on Sabouraud dextrose agar with chloramphenicol at 25°C and 37°C. Anti-fungal susceptibility test was done by disk diffusion method using medium Muller-Hilton agar with glucose and methylene blue. All the patients were treated with the removal of fungal debris with topical clotrimazole instillation and were followed up after 7 days. Non-responders were treated with topical itraconazole. Results: Male-to-female ratio in study participants was 1:1.3. The most frequently observed risk factor was the instillation of oil in the ear (20%). Ear itching was the commonest symptom (70.9%). Positive fungal growth was seen in 203 (88.3%) cases. Aspergillus was present in 185 (91.1%) cases, followed by Candida in 18 (8.9%) cases. Voriconazole had the highest sensitivity followed by itraconazole. 167 (83.3%) patients responded to primary treatment (topical Clotrimazole), 36 (16.7%) patients were non-responders. Conclusion: All the clinically suspected patients should be evaluated for fungal positivity, which may further guide in the selection of appropriate anti-fungal agents. Awareness regarding the predisposing factors for otomycosis in the general population may result in a significant drop in the incidence of otomycosis.

Genetic diversity and drug sensitivity profile of Mycobacterium tuberculosis among children in Ethiopia.

Worldwide, tuberculosis (TB) affects about one million children every year. The burden of the disease is higher in developing countries. However, there is limited information on the lineages and drug sensitivity patterns of Mycobacterium tuberculosis (M. tuberculosis) infecting children in these countries, including Ethiopia. Thus, this study aimed to characterize the different lineages of the M. tuberculosis complex causing childhood pulmonary tuberculosis and evaluate the drug-sensitivity patterns to the first-line anti-TB drugs. A total of 54 stored cultures were used in this study. The region of difference 9 (RD9) based polymerase chain reaction (PCR) and spoligotyping were employed for the identification of the isolates at the species and lineages level respectively. Lineage identification was done by using the pre-existing database. Identification of clustering of the spoligotype patterns was by using the SPOLIDB3-based model. The result was retrieved by the most probable family format. Furthermore, the phenotypic, and genotypic drug-sensitivity test (DST) was performed using Mycobacterium Growth Indicator Tube (MGIT™ 960) and GenoTypeMTBDRplus assay respectively. Data analysis was done using SPSS version 27 software. Spoligotyping produced 39 interpretable results for M. tuberculosis. The majority (74.4%) of them were clustered into 7 groups, while the rest (25.6%) were single. The Euro-American (EA) lineage was the predominant lineage (64.1%) followed by the East-African Indian (EAI) (30.8%) and M. Africanum (5.1%) lineages. The most predominant subtypes were SIT37 (15.4%), SIT149 (12.8%), SIT25 (7.7%), and SIT53 (7.7%). Furthermore, of the identified SITs, T1 and CAS families consisted of 38.5% and 28.2% of the lineages respectively. Drug susceptibility was 91.9% by phenotypic method and 97.4% by molecular assay. The overall prevalence of any resistance was 7.8% and there was a single MDR-TB. Many of the isolates belong to the modern lineages (Euro American) representing the most common circulating strains in the country. More importantly, despites the tiny isolates tested, drug resistance is low. To fully describe the molecular epidemiology of MTBC lineages in children, we recommend a prospective large-scale study.

A study of etiopathogenesis, clinical and laboratory profile evaluation of typhoid fever in paediatric age group in Bijapur

Background: Salmonella typhi causes typhoid illness. In India, it is a huge public health issue. Several impoverished nations have endemic typhoid fever. The wide range of clinical symptoms of typhoid fever makes diagnosis difficult. The purpose of this study was to better understand the vast range of clinical symptoms, comorbidities, and drug sensitivity patterns associated with typhoid fever in children. Methods: From March 2021 to October 2022, 100 children hospitalised to the paediatric unit at Al-Ameen Medical College and Hospital in Bijapur with proven typhoid fever were included in the study. Age, gender, presenting complaint, laboratory tests, and antibiotic sensitivity pattern are all gathered and analysed in each patient. Results: There were 58 men (58%) and 42 females (42%) among the 100 cases. The most typical age range was 5 to 15 years. Fever was the most prevalent symptom, occurring in 100% of patients, headache (80%), coated tongue (100%), vomiting (45%), and abdominal pain (70%). In 68% of cases, the most prevalent indication was a toxic look, and hepatomegaly in 44%. In 28% of instances, leukocytopenia was discovered. In every case, the blood culture proved positive. The length of hospital stay ranged from 3 to 10 days. There was no recorded death. Conclusions: Unhygienic habits and consuming meals from the outside are the main causes of typhoid fever. By raising people's understanding of disease transmission and its many preventative methods, this serious public health issue can be resolved.

Molecular typing and drug sensitivity profiles of M. Tuberculosis isolated from refugees residing in Ethiopia

BackgroundRefugees in developing countries have poor access to Tuberculosis (TB) care and control services. The understanding of genetic diversity and drug sensitivity patterns of M. tuberculosis (MTB) is important for the TB control program. However, there is no evidence that shows the drug sensitivity profiles and genetic diversity of MTB circulating among refugees residing in Ethiopia. This study aimed to investigate the genetic diversity of MTB strains and lineages, and to identify the drug sensitivity profiles of MTB isolated from refugees residing in Ethiopia. MethodsA cross-sectional study was conducted among 68 MTB positive cases isolated from presumptive TB refugees from February to August 2021. Data and samples were collected in the refugee camp clinics and both rapid TB Ag detection and region of difference (RD)-9 deletion typing were used to confirm the MTBs. Drug susceptibility test (DST) and molecular typing were done using Mycobacterium Growth Indicator Tube (MGIT) method and spoligotyping respectively. ResultsDST and spoligotyping results were available for all 68 isolates. The isolates were grouped into 25 spoligotype patterns, which consisted of 1–31 isolates with 36.8% strain diversity. The international shared type (SIT)25 was predominant spoligotype pattern consisting of 31 (45.6%) isolates, followed by SIT24 comprising 5 (7.4%) isolates. Further investigation showed that 64.7% (44/68) of the isolates were belonged to CAS1-Delhi family and 75% (51/68) of the isolates were belonged to lineage(L)-3. Multi-drug resistance (MDR)-TB was observed only in one isolate (1.5%) for first-line anti-TB drugs and the highest level of mono-resistance, 5.9% (4/68), was observed for PZA(Pyrazinamide). Mono-resistance was observed in 2.9 % (2/68) and while 97.0% (66/68) of the MTB positive cases were susceptible to the second-line anti-TB drugs. ConclusionThe findings are useful evidence for the TB screening, treatment and control in refugee populations and surrounding communities in Ethiopia.