Ceftobiprole and Cefiderocol for Patients on Extracorporeal Membrane Oxygenation: The Role of Therapeutic Drug Monitoring.

INTRODUCTION: Limited data exist on therapeutic ranges for newer antimicrobials in the critically ill, with few pharmacokinetic studies including patients undergoing renal replacement therapy or extracorporeal membrane oxygenation (ECMO). CASE REPRESENTATION: These interventions can potentially alter the pharmacokinetic profile of antibiotics, resulting in therapeutic failures, antimicrobial resistance, or increased toxicity. In this report, we present two ECMO patients treated with cefiderocol and ceftobiprole, where therapeutic drug monitoring (TDM) aided in the successful treatment of severe infections. Antibiotic trough concentrations in both cases were consistent with previously reported therapeutic levels in critically ill and ECMO patients, meeting minimal inhibitory concentrations recommended by the European Committee on Antimicrobial Susceptibility Testing for the respective pathogens. CONCLUSION: Treatment might be suboptimal if doses are not adjusted based on physicochemical properties and extracorporeal support. In an era marked by highly resistant pathogens, these cases highlight the importance of timely access to real-time TDM for optimizing and individualizing antimicrobial treatment.

The Therapeutic Potential of Targeting Tumor Microenvironment and Modulation of Immunotherapy in Gastrointestinal Cancer.

Immunotherapy, as a novel treatment approach for various disorders, including cancers, is designed to either stimulate or suppress the immune system with high speci-ficity. The recent achievements of this therapy in clinical trials are set to transform tradi-tional treatment methods. Furthermore, it holds promise for enhancing the survival rates of patients suffering from both metastatic cancers and primary stages. Gastrointestinal Cancers (GI) account for 26% of global incidence and 35% of worldwide deaths. Treat-ment can be carried out using targeted immunotherapy in these cancers. If the tiers are superior, improvement could require more enterprise. On account that the function of immunotherapy in GI has been so promising, solely in sufferers with severe metastatic levels, within the literature, the immune checkpoint inhibitors in cancer immunotherapy of GI cancers, chimeric antigen receptor T-cell (vehicle-T), modulators of the tumor mi-croenvironment, and drug resistance mechanisms in immunotherapy as an effective treatment approach to GI cancers along with colon, pancreas, gastric, and esophageal cancers have been addressed. This review provides an overview of FDA-approved im-munotherapy drugs and ongoing preclinical developments. Additionally, we offer in-sights into the future of immunotherapy for GI cancer patients, addressing the associated challenges.

A genome-wide CRISPR/Cas9 knockout screen identifies SEMA3F gene for resistance to cyclin-dependent kinase 4 and 6 inhibitors in breast cancer.

BACKGROUND: Palbociclib is a cell-cycle targeted small molecule agent used as one of the standards of care in combination with endocrine therapy for patients with hormone receptor (HR)-positive, human epidermal growth factor receptor 2 (HER2)-negative advanced breast cancer. Although several gene alterations such as loss of Rb gene and amplification of p16 gene are known to be conventional resistance mechanisms to cyclin-dependent kinase 4 and 6 (CDK4/6) inhibitors, the comprehensive landscape of resistance is not yet fully elucidated. The purpose of this study is to identify the novel resistant genes to the CDK4/6 inhibitors in HR-positive HER2-negative breast cancer. METHODS: The whole genome knockout screen using CRISPR/Cas9 genome editing was conducted in MCF7 to identify resistant genes to palbociclib. The candidate genes for resistance were selected by NGS analysis and GSEA analysis and validated by cell viability assay and mouse xenograft models. RESULTS: We identified eight genes including RET, TIRAP, GNRH1, SEMA3F, SEMA5A, GATA4, NOD1, SSTR1 as candidate genes from the whole genome knockout screen. Among those, knockdown of SEMA3F by siRNA significantly and consistently increased the cell viability in the presence of CDK4/6 inhibitors in vitro and in vivo. Furthermore, the level of p-Rb was maintained in the palbociclib treated SEMA3F-downregulated cells, indicating that the resistance is driven by increased activity of cyclin kinases. CONCLUSION: Our observation provided the first evidence of SEMA3F as a regulator of sensitivity to CDK4/6 inhibitors in breast cancer. The detailed mechanisms of resistance deserve further functional studies to develop the better strategy to overcome resistance in CDK4/6 inhibitors.

A Phase II Clinical Study on Apatinib Plus Vinorelbine in Refractory HER2-Negative Breast Cancer and its Metabolic Implications of Drug Resistance.

BACKGROUND: Apatinib, a tyrosine-kinase inhibitor that targets the vascular endothelial growth factor receptor 2, contributes to the inhibition of angiogenesis. Vinorelbine, a semisyn-thetic vinca alkaloid, primarily inhibits metaphase mitosis of cancer cells through its interactions with tubulin. This study aimed to evaluate whether apatinib combined with vinorelbine was ef-fective and safe for refractory human epidermal growth factor receptor 2 (HER2)-negative breast cancer patients who failed taxanes and/or anthracycline and analyze the possible mechanism of drug resistance through metabolomic analysis. METHODS: Eligible patients were HER2-negative, inoperable, locally advanced, or metastatic breast cancer patients who progressed after at least one chemotherapy regimen in this present prospective phase II study. Patients took oral apatinib (250-500 mg/day) plus intravenous infusion of vinorelbine (25 mg/m2 on day 1, day 8 at 3-week intervals). Objective response rate (ORR) was our primary endpoint, while disease control rate (DCR), overall survival (OS), progression-free survival (PFS), and toxicity were our secondary endpoints. The exploratory purpose was to identify biomarkers or drug resistance mechanisms through metabolomics changes before and after the combination therapy. RESULTS: Between September, 2019 and June, 2022, a total of 34 patients were included. ORR and DCR were 32.4% (11/34) and 85.3% (29/34), respectively. The median PFS was 5.0 months (95% CI, 3.766-6.234), while the median OS was 13.0 months (95% CI, 8.714-17.286). Side effects included hematologic toxicity, gastrointestinal reaction, and sinus tachycardia, which were mild to moderate. The mainly disturbed metabolic pathways were the cAMP signaling pathway, the alanine/aspartate/glutamate metabolism, the central carbon metabolism in cancer, the beta-alanine metabolism, the butanoate metabolism, and the glyoxylate and dicarboxylate metabolism, which may lead to the resistance of patients to this combination therapy. CONCLUSION: Apatinib combined with vinorelbine is effective and safe in patients with locally advanced or metastatic refractory HER2-negative breast cancer. The findings of this study con-tribute to a better understanding of the metabolic effect of apatinib and vinorelbine therapy.

Increasing rate of non-Candida albicans yeasts and fluconazole resistance in yeast isolates from women with recurrent vulvovaginal candidiasis in Leeds, United Kingdom.

OBJECTIVES: Azoles have been the mainstay of recurrent vulvovaginal candidiasis (RVVC) for many years. Because of a recent anecdotal increase in non-Candida albicans yeasts (NCAY) and azole-resistant C. albicans cases, their prevalence was calculated from cultures for yeasts in women with complicated/RVVC over 3 years. METHODS: Retrospective data search of vaginal cultures from adult women in Leeds, UK between April 2018 and March 2021 was conducted. Samples with clinical details of complicated/RVVC had full yeast identification and antifungal susceptibility performed. Differences in prevalence between 12-month periods were determined using χ2 tests. RESULTS: Over the 3 years, cultures were performed on 5461 vaginal samples from women with clinical information indicating they had complicated/RVVC, RVVC, with 1828 (33.5%) growing yeasts.Over 85% of yeasts each year were C. albicans, however the proportion declined yearly with an increase in NCAY species. Nakaseomyces glabrata was the most frequent NCAY species isolated, increasing from 2.8% in 2018-19 to 6.8% in 2020-21. Total NCAY species increased from 6.0% in 2018-19 to 12.6% in 2020-21. Fluconazole-sensitive dose-dependant (SDD) and resistant isolates increased from 3.5% in 2018-19 to 7.7% in 2019-20 and 9.6% in 2020-21. Most resistance was in C. albicans and the majority of cases were seen in primary care. Most fluconazole non-sensitive isolates were either SDD or resistant to itraconazole (77% and 23%, respectively) and were intermediate or resistant to voriconazole (36.4% and 60%, respectively). CONCLUSION: There was a significant increase in the prevalence of NCAY and fluconazole-resistant C. albicans in complicated/RVVC cultures over these 3 years. Successful treatment of such cases can be very challenging. The exact reasons for this increase remain unclear but it follows a policy change that encouraged a clinical diagnosis and empirical treatment of vulvovaginal candidiasis, rather than fungal culture, in primary care.

Modulations of the skin microbiome in skin disorders: A narrative review from a wound care perspective.

The cutaneous microbiome represents a highly dynamic community of bacteria, fungi and viruses. Scientific evidence, particularly from the last two decades, has revealed that these organisms are far from being inconsequential microscopic hitchhikers on the human body, nor are they all opportunistic pathogens waiting for the chance to penetrate the skin barrier and cause infection. In this review, we will describe how dermatological diseases have been found to be associated with disruptions and imbalances in the skin microbiome and how this new evidence had shaped the diagnosis and clinical practice relating to these disorders. We will identify the microbial agents which have been found to directly exacerbate skin diseases, as well as those which can ameliorate many of the symptoms associated with dermatological disorders. Furthermore, we will discuss the studies which suggest that bacteriotherapy, either by topical use of probiotics or by bacteria-derived compounds, can rectify skin microbial imbalances, thereby offering a promising alternative to antibiotic treatment and reducing the risks of antibiotic resistance.

Nanobiotechnology boosts ferroptosis: opportunities and challenges.

Ferroptosis, distinct from apoptosis, necrosis, and autophagy, is a unique type of cell death driven by iron-dependent phospholipid peroxidation. Since ferroptosis was defined in 2012, it has received widespread attention from researchers worldwide. From a biochemical perspective, the regulation of ferroptosis is strongly associated with cellular metabolism, primarily including iron metabolism, lipid metabolism, and redox metabolism. The distinctive regulatory mechanism of ferroptosis holds great potential for overcoming drug resistance-a major challenge in treating cancer. The considerable role of nanobiotechnology in disease treatment has been widely reported, but further and more systematic discussion on how nanobiotechnology enhances the therapeutic efficacy on ferroptosis-associated diseases still needs to be improved. Moreover, while the exciting therapeutic potential of ferroptosis in cancer has been relatively well summarized, its applications in other diseases, such as neurodegenerative diseases, cardiovascular and cerebrovascular diseases, and kidney disease, remain underreported. Consequently, it is necessary to fill these gaps to further complete the applications of nanobiotechnology in ferroptosis. In this review, we provide an extensive introduction to the background of ferroptosis and elaborate its regulatory network. Subsequently, we discuss the various advantages of combining nanobiotechnology with ferroptosis to enhance therapeutic efficacy and reduce the side effects of ferroptosis-associated diseases. Finally, we analyze and discuss the feasibility of nanobiotechnology and ferroptosis in improving clinical treatment outcomes based on clinical needs, as well as the current limitations and future directions of nanobiotechnology in the applications of ferroptosis, which will not only provide significant guidance for the clinical applications of ferroptosis and nanobiotechnology but also accelerate their clinical translations.

The antimicrobial resistance landscape of slaughterhouses in western Kenya: A microbiological case study.

Slaughterhouses may be hotspots for the transmission of antimicrobial resistant (AMR) pathogens. To obtain information on the AMR landscape in Kenyan slaughterhouses, we collected swabs of the environment, animal carcasses, and workers. Bacterial isolates were identified in 101/193 (52.3 %) samples, and most showed resistance to streptomycin (68.7 %), ampicillin (48.7 %), and tetracycline (42.5 %). Multi drug resistance was exhibited by 35/80 isolates (43.8 %; 95 % CI: 33.2-54.9 %), while Extended Spectrum Beta Lactamase was expressed in 5/80 isolates (6.3 %; 95 % CI: 2.6-14.3 %). These findings illustrate the presence of resistant bacteria throughout the slaughterhouse environment, posing a risk to workers and meat consumers and highlighting the need for an integrated surveillance system along the food chain.

Primary Caregiver Knowledge about Self-Medication of Antibiotics in Children Aged 0-12 Years.

Globally, it is estimated that more than 50% of antibiotics are obtained without a prescription. The main purpose of this study is to determine the knowledge and practice of primary caregivers about self-medication in children with antibiotics, as studies on self-medication is lacking in India, also, it will help is assessing parents' knowledge and attitude towards self medication. This cross-sectional study conducted in the urban community of Shastri Nagar, Patna, aimed to evaluated antibiotic use in children aged 0-12. From January 2023 to March 2023, 173 caregivers were randomly selected through house visits. Data collection used a pre-tested questionnaire, ensuring confidentiality. In this study of 173 participants, caregivers in an urban community demonstrated varying knowledge regarding antibiotic use in children. Mothers and post-graduates possessed better awareness of antibiotic consequences. Fathers exhibited better understanding of side effects. Knowledge on antibiotics' action was seen among mothers, those aged 30-39, with family income of Rs. 20,000-40,000 and those with family members in medical field. Fathers had more incorrect beliefs about antibiotics treating viral infections. Common conditions for self-medication included cough/cold, fever and diarrhea, with hospitals being the primary source of antibiotics. Majority obtained information from pharmacies but awareness about antibiotic course completion and versatility was limited. Caregivers' antibiotic knowledge varied; mothers and post-graduates were more aware, while fathers understood side effects better.

Insights into existing and futuristic treatment approach for chronic myeloid leukaemia.

Oncogenes play a crucial part in human cancer development, and when particular drugs obstruct the proteins produced by these oncogenes, the tumoural process can be ceased. For instance, in chronic myeloid leukaemia (CML), all pathological traits are associated with a single oncogene, BCR-ABL1. CML is a triphasic cancerous disorder of haematopoietic stem cells, marked by a balanced translocation between chromosomes 9 and 22, leading to the genesis of a Philadelphia chromosome encompassing the BCR-ABL1 fusion gene. This fusion oncogene further produces a constitutive active tyrosine kinase protein, enhancing the downstream signalling pathways and constitutes cancer. The treatment for CML has been entirely altered from chemotherapy and immunotherapy to targeted therapy with the emergence of tyrosine kinase inhibitors (TKIs) which inhibit BCR-ABL1 kinase activity. However, the inhibitory mechanism of TKIs is constrained by BCR-ABL1 dependent and independent resistance mechanisms, prompting the exploration of novel therapeutics through extensive clinical trials to develop next-generation drugs with enhanced potency. The persistent challenges posed by CML have motivated researchers to seek innovative strategies for its eradication, such as the application of the genome editing tool CRISPR/Cas9. This review provides insights into existing CML diagnoses, treatment modalities, resistance mechanisms, drugs under trial phases and new potential therapeutic drugs. Furthermore, the review looks ahead to a visionary perspective wherein the CRISPR/Cas9 approach holds the potential to evolve into a prospective curative measure for CML.

Clinical Infection Characteristics and Drug Resistance of Group B Streptococcus in Chengdu, China.

OBJECTIVE: To investigate the clinical infection characteristics and antibiotic resistance of Group B Streptococcus (Streptococcus agalactiae, GBS) in Chengdu, China, from 2019 to 2021, as well as to provide data to support rational clinical drug use. METHODS: This was a retrospective study to collect 203 culture-positive GBS strains isolated from January 2019 to December 2021 in Chengdu, China, all of which were identified by the VITEK 2 Compact automated microbial Bacterial identification instrument. Data were derived using WHONET 5.6 software. The sample type and ward distribution were counted. Pregnant women and newborns were screened from the original data and their pregnancy outcomes were calculated respectively. RESULTS: GBS strains were mainly concentrated in obstetrics and neonatology departments, accounting for 40.9% and 33.5%. The types of specimens were mainly vaginal secretions, amniotic fluid and sputum, accounting for 25.6%, 26.1% and 18.7%, respectively. Chorioamnionitis, premature rupture of membranes and preterm delivery occurred mainly in pregnant women after infection, accounting for 44.4%, 31.5% and 24.1%. Neonates, on the other hand, were mainly diagnosed with neonatal pneumonia, neonatal sepsis, respiratory failure and septic meningitis, accounting for 91.8%, 61.2%, 44.9% and 16.3% of all positive neonates. 840 pregnant women were screened for GBS colonization from 2019-2021, and a total of 108 GBS positive pregnant women were identified, with a GBS colonization rate of 12.9%. A total of 9 neonates from 108 GBS positive pregnant women developed early-onset disease. The morbidity in neonates was 8.3%. No strains resistant to penicillin and ampicillin were found, while the resistance rates of tetracycline and clindamycin were higher than 50%, respectively 60.1% and 53.2%. CONCLUSION: GBS infection mainly affected pregnant women and newborns in Chengdu, China, which can lead to adverse maternal and infant outcomes. Attention should be paid to strengthening general screening of GBS in perinatal urogenital secretions and the prevention strategy of IAP (intrapartum antibiotic prophylaxis). Antimicrobial therapy should be administered with appropriate antibiotics. Penicillin was still the first line drug for the treatment of GBS. These initiatives were important to reduce mother-to-child transmission and neonatal infections.

Disinfectants efficacy in reducing pathogens related to health-care infection associated in universities hospitals of Gorgan, North of Iran.

BACKGROUND: Disinfection has a fundamental role in the control of pathogens in the hospital environment. This study was designed to assess the efficacy and functional impact of disinfectants in reducing pathogens related to healthcare associated infections (HAIs) in hospitals. METHODS: This observation study was conducted at three university hospitals in Gorgan, Iran, from May to Oct 2023. The data including used disinfectants and microbiological examination were obtained from the infection control unit of each hospital. RESULTS: The results showed that a variety of disinfectants from intermediate to high levels were employed in accordance with the World Health Organization (WHO) protocols. The microbial result revealed that 31.6% (286 out of 906) of the sample had at least one microorganism. Among identified organisms, Bacillus spp. were the predominant species followed by Staphylococcus epidermis, fungus genera, Enterobacter spp., Enterococcus spp., Pseudomonas spp., Escherichia coli, Alcaligenes spp., Staphylococcus aureus, Citrobacter spp., Corynebacterium spp., Klebsiella spp., Acinetobacter spp., Micrococcus spp., Staphylococcus saprophyticus, and Serratias spp. The highest prevalence rates of microorganisms were observed in the wards of ICU, emergency, internal medicine, and women's ward. The chi-square test revealed a significant relationship between the presence of organisms and hospital wards (P < 0.05). CONCLUSION: The presence of pathogens indicates a defect in the disinfection process, probably due to both little attention to disinfection protocols and multidrug resistance. It is not yet possible to eliminate pathogens from the hospital environment, but it can be minimized by education intervention, standardizing disinfecting processes, and monitoring by the infection control committee.

High frequency of artemisinin partial resistance mutations in the Great Lakes region revealed through rapid pooled deep sequencing.

BACKGROUND: In Africa, the first Plasmodium falciparum artemisinin partial resistance mutation, was Kelch13 (K13) 561H - detected and validated at appreciable frequency in Rwanda in 2014. Surveillance to better define the extent of the emergence in Rwanda and neighboring countries is critical. METHODS: We used novel liquid blood drop preservation combined with pooled sequencing to provide cost-effective rapid assessment of resistance mutation frequencies at multiple collection sites across Rwanda and neighboring regions in Uganda, Tanzania, and the Democratic Republic of the Congo (DRC). Malaria-positive samples (n=5,465) from 39 health facilities collected between May 2022 and March 2023 were sequenced in 199 pools. RESULTS: In Rwanda, K13 561H and 675V were detected in 90% and 65% of sites with an average frequency of 19.0% (0-54.5%) and 5.0% (0-35.5%), respectively. In Tanzania, 561H had high frequency in multiple sites. 561H appeared at 1.6% in Uganda. 561H was absent from the DRC, although 675V was seen at low frequency. Concerningly candidate mutations were observed: 441L, 449A, and 469F co-occurred with validated mutations suggesting they are arising under the same pressures. Other markers for decreased susceptibility to artemether-lumefantrine are common: P. falciparum multidrug resistance protein 1 N86 at 98.0% (63.3-100%) and 184F at 47.0% (0-94.3%) and P. falciparum chloroquine resistance transporter 76T at 14.7% (0-58.6%). Additionally, sulfadoxine-pyrimethamine-associated mutations show high frequencies. CONCLUSION: K13 mutations are rapidly expanding in the region further endangering control efforts with the potential of engendering partner drug resistance.

Variability of macrolide-resistant profile in Mycobacterium avium complex pulmonary disease.

This single-center retrospective study aimed to analyze the variability of macrolide resistance (MR) in 68 patients with Mycobacterium avium complex pulmonary disease. Among 25 patients treated without macrolides, 13 (52%) reverted to macrolide-susceptible (MS) profiles. Only one (2%) of 43 patients who continued macrolide treatment showed this change. We compared 30 MR isolates with recent specimens. Among them, seven shifted to MS (five attributed to clonally related strains; two resulting from reinfection or polyclonal infection).

CASPER: A Phase I trial combining calaspargase pegol-mnkl and cobimetinib in pancreatic cancer.

Asparagine synthetase (ASNS) catalyzes the biosynthesis of asparagine from aspartate and glutamine. Cells lacking ASNS, however, are auxotrophic for asparagine. Use of L-asparaginase to promote asparagine starvation in solid tumors with low ASNS levels, such as pancreatic ductal adenocarcinoma (PDAC), is a rationale treatment strategy. However, tumor cell resistance to L-asparaginase has limited its clinical utility. Our preclinical studies show that RAS/MAPK signaling circumvents L-asparaginase-induced tumor killing, but L-asparaginase and MEK inhibition potentiated tumor killing; suggesting that this combination may provide meaningful clinical benefit to patients with PDAC. This Phase I trial (NCT05034627) will evaluate the safety and tolerability of the MEK inhibitor, cobimetinib, in combination with pegylated L-asparaginase, L calaspargase pegol-mknl, in patients with locally-advanced or metastatic PDAC.

[Box: see text].

Paeoniae radix overcomes resistance to EGFR-TKIs via aurora B pathway suppression in lung adenocarcinoma.

Targeted therapies using epidermal growth factor receptor (EGFR) inhibitors have markedly improved survival rates and quality of life for patients with EGFR-mutant lung adenocarcinoma (LUAD). Despite these advancements, resistance to EGFR inhibitors remains a significant challenge, limiting the overall effectiveness of the treatment. This study explored the synergistic effects of combining Paeoniae Radix (PR) with first-generation EGFR-tyrosine kinase inhibitors (TKIs), erlotinib and gefitinib, to overcome this resistance. Transcriptomic analysis of EGFR-mutant LUAD cell lines revealed that PR treatment could potentially reverse the gene signatures associated with resistance to EGFR-TKIs, primarily through the suppression of the Aurora B pathway. Experimental validation demonstrated that combining PR with erlotinib and gefitinib enhanced drug responsiveness by inhibiting Aurora kinase activity and inducing apoptosis in LUAD cells. Additionally, gene expression changes confirmed these combined effects, with the suppression of the Aurora B pathway and upregulation of the apoptotic pathway, which was accompanied by increased expression of multiple pro-apoptotic genes. Our findings contribute to the development of natural product-based therapeutic strategies to mitigate drug resistance in LUAD.

Parasitological prevalence of scabies and secondary bacterial infections among scabies suspected patients at Borumeda General Hospital, Northeast Ethiopia.

BACKGROUND: Scabies disproportionately affects people in resource-poor areas. Clinical diagnosis risks misdiagnosis due to resemblance to other skin diseases, but laboratory confirmation improves accuracy. Scabies allow for secondary bacterial infections. Associated bacteria exacerbate scabies and antibiotic resistance. Ethiopian scabies diagnosis relies solely on clinical exams without confirming parasites or investigating secondary bacterial infections. This study aims to identify parasites via scraping, isolate secondary bacteria, and evaluate their antibiotic susceptibility profiles. METHODS: A hospital based cross-sectional study was conducted from September 2022 to July 2023 among scabies suspected patients who visited the dermatology clinic at Borumeda General Hospital in Northeast Ethiopia. A systematic random sampling technique was used to select 422 study participants. Socio-demographic, hygiene, and clinical characteristics data were collected via face-to-face interviews and observation. Skin scrapings for parasitological investigations and swab samples for microbiological investigations were collected and transported for analysis and drug susceptibility testing. Descriptive statistics and logistic regression analysis were employed to assess risk factors. RESULT: Among 422 skin scraping samples, 156 (37.0%) cases of scabies were microscopically confirmed. Bed-sharing and having contact history were independent predictors of microscopically confirmed scabies. The prevalence of secondary bacterial infections among scabies-confirmed patients was 35.9% (56/156). The most prevalent bacterial species were Staphylococcus aureus, coagulase-negative staphylococci, and Streptococcus pyogenes. Tetracycline for Gram-positive bacteria and ampicillin for Gram-negative bacteria showed the highest rate of resistance. In both Gram-positive and Gram-negative strains, multidrug resistance was also observed. CONCLUSION: This study found that over one-third of skin scrapings tested positive for scabies. Additionally, more than one-third of scabies cases were complicated by secondary bacterial infections. Improved scabies diagnosis and consideration of secondary bacterial infections are important for better controlling this neglected tropical disease.

Wastewater-based surveillance of Vibrio cholerae: Molecular insights on biofilm regulatory diguanylate cyclases, virulence factors and antibiotic resistance patterns.

Vibrio cholerae is an inherent inhabitant of aquatic ecosystems. The Indian state of West Bengal, especially the Gangetic delta region is the highest cholera affected region and is considered as the hub of Asiatic cholera. V. cholerae were isolated from publicly accessible wastewater of Midnapore, West Bengal, India. Serotyping determined all isolates to be of non-O1/non-O139 serogroups. Moderate biofilm-forming abilities were noticed in most of the isolates (74.7 %) while, high biofilm formation was recorded for only 6.3 % isolates and 19 % of isolates exhibited low/non-biofilm-forming abilities. PCR-based screening of crucial diguanylate cyclases (DGCs) involved in cyclic-di-GMP-mediated biofilm signaling was performed. cdgH and cdgM were the most abundant DGCs among 93.7 % and 91.5 % of isolates, respectively. Other important DGCs, i.e., cdgK, cdgA, cdgL, and vpvC were present in 84 %, 75.5 %, 72 % and 68 % of isolates, respectively. Besides, the non-O1/non-O139 isolates were screened for the occurrence of virulence factor encoding genes. Moreover, among these non-O1/non-O139 isolates, two strains (3.17 %) harbored both ctxA and ctxB genes, which encode the cholera toxin associated with epidemic cholera. ompU was the most prevalent virulence factor, present in 24.8 % of isolates. Other virulence factors like, zot and st were found in 4.7 % and 9.5 % of isolates. Genes encoding tcp and ace were found to be PCR-negative for the isolates. Additionally, crucial virulence factor regulators, toxT, toxR and hapR were found to be PCR-positive in all the isolates. Antibiotic resistance patterns displayed further vulnerabilities with decreased sensitivity towards commonly used antibiotics with multiple antibiotic resistance index ranging between 0.37 and 0.62. The presence of cholera toxin-encoding multi-drug resistant (MDR) V. cholerae strains in environmental settings is alarming. High occurrence of DGCs are considered to encourage further investigations to use them as alternative therapeutic targets against MDR cholera pathogen due to their unique presence in bacterial systems.

Another decade of antimalarial drug discovery: New targets, tools and molecules.

Malaria remains a devastating but preventable infectious disease that disproportionately affects the African continent. Emerging resistance to current frontline therapies means that not only are new treatments urgently required, but also novel validated antimalarial targets to circumvent cross-resistance. Fortunately, tremendous efforts have been made by the global drug discovery community over the past decade. In this chapter, we will highlight some of the antimalarial drug discovery and development programmes currently underway across the globe, charting progress in the identification of new targets and the development of new classes of drugs to prosecute them. These efforts have been complemented by the development of valuable tools to accelerate target validation such as the NOD scid gamma (NSG) humanized mouse efficacy model and progress in predictive modelling and open-source software. Among the medicinal chemistry programmes that have been conducted over the past decade are those targeting Plasmodium falciparum ATPase4 (ATP4) and acetyl-CoA synthetase (AcAS) as well as proteins disrupting parasite protein translation such as the aminoacyl-tRNA synthetases (aaRSs) and eukaryotic elongation factor 2 (eEF2). The benefits and challenges of targeting Plasmodium kinases will be examined, with a focus on Plasmodium cyclic GMP-dependent protein kinase (PKG), cyclin-dependent-like protein kinase 3 (CLK3) and phosphatidylinositol 4-kinase (PI4K). The chapter concludes with a survey of incipient drug discovery centres in Africa and acknowledges the value of recent international meetings in galvanizing and uniting the antimalarial drug discovery community.

Widely-targeted in silico and in vitro evaluation of veratrum alkaloid analogs as FAK inhibitors and dual targeting of FAK and Hh/SMO pathways for cancer therapy: A critical analysis.

Focal Adhesive Kinase (FAK), a key player in aggressive cancers, mediates signals crucial for progression, invasion, and metastasis. Despite advances in targeted therapies, drug resistance is still a challenge, and survival rates remain low, particularly for late-stage patients, emphasizing the need for innovative cancer therapeutics. Cyclopamine, a veratrum alkaloid, has shown promising anti-tumor properties, but the search for more potent analogs with enhanced affinity for the biological target continues. This study employs a hybrid virtual screening approach combining pharmacophore model-based virtual screening (PB-VS) and docking-based virtual screening (DB-VS) to identify potential inhibitors of the FAK catalytic domain. PB-VS on the PubChem database yielded a set of hits, which were then docked with the FAK catalytic domain in two stages (1st and 2nd DB-VS). Hits were ranked based on docking scores and interactions with the active site. The top three compounds underwent molecular dynamics simulations, alongside two control compounds (SMO inhibitor(s) and FAK inhibitor(s)), to assess stability through RMSD, RMSF, Rg, and SASA analyses. ADMET properties were evaluated, and compounds were filtered based on drug-likeness criteria. Molecular dynamics simulations demonstrated the stability of compounds when complexed with the FAK catalytic domain. Compounds 16 (-25 kcal/mol), 88 (-27.47 kcal/mol), and 87 (-18.94 kcal/mol) exhibited comparable docking scores, interaction profiles, stability, and binding energies, indicating their potential as lead candidates. However, further validation and optimization through quantitative structure-activity relationship (QSAR) studies are essential to refine their efficacy and therapeutic potential. The in vitro cell-based assay demonstrated that compound 101PF, a FAK inhibitor, significantly inhibited the proliferation and migration of A549 cells. However, the results regarding the combined effects of FAK and SMO inhibitors were inconclusive, highlighting the need for further investigation. This study contributes to developing more effective anti-cancer drugs by improving the understanding of potential cyclopamine-based veratrum alkaloid analogs with enhanced interactions with the FAK catalytic domain.