Antibiotic Resistant Biofilms and the Quest for Novel Therapeutic Strategies.

Antimicrobial resistance (AMR) is one of the major leading causes of death around the globe. Present treatment pipelines are insufficient to overcome the critical situation. Prominent biofilm forming human pathogens which can thrive in infection sites using adaptive features results in biofilm persistence. Considering the present scenario, prudential investigations into the mechanisms of resistance target them to improve antibiotic efficacy is required. Regarding this, developing newer and effective treatment options using edge cutting technologies in medical research is the need of time. The reasons underlying the adaptive features in biofilm persistence have been centred on different metabolic and physiological aspects. The high tolerance levels against antibiotics direct researchers to search for novel bioactive molecules that can help combat the problem. In view of this, the present review outlines the focuses on an opportunity of different strategies which are in testing pipeline can thus be developed into products ready to use.

Increased electrode impedance as an indicator for early detection of deep brain stimulation (DBS) hardware Infection: Clinical experience and in vitro study.

BACKGROUND: When deep brain stimulation (DBS) infections are identified, they are often too advanced to treat without complete hardware removal. New objective markers to promptly identify DBS infections are needed. We present a patient with GPi (globus pallidus interna) DBS for dystonia, where the electrode impedance unexpectedly increased 3-months post-operatively, followed by serologic and hematologic markers of inflammation at 6-months, prompting explantation surgery. We recreated these conditions in a laboratory environment to analyze the pattern of changing of electrical impedance across the contacts of a DBS lead following Staphylococcus biofilm formation. METHODS: A stainless-steel culture chamber containing 1 % brain heart infusion agar was used. A DBS electrode was dipped in peptone water containing a strain of S. aureus and subsequently introduced into the chamber. The apparatus was incubated at 37 °C for 6 days. Impedance was measured at 24hr intervals. A control experiment without S. Aureus inoculation was used to determine changes in impedance over a period of 6-days. RESULTS: The mean monopolar impedance on day-1 was 751.8 ± 23.8 Ω and on day-3 was 1004.8 ± 68.7 Ω, a 33.7 % rise (p = 0.007). A faint biofilm formation could be seen around the DBS lead by day-2 and florid growth by day-3. After addition of the linezolid solution, a 15.9 % decrease in monopolar impedance was observed from day 3-6 (p = 0.003). CONCLUSION: This study gives insight into impedance trends following a hardware infection in DBS. Increased impedance outside expected norms may be valuable for early prediction of infection. Furthermore, timely management using antibiotics might reduce the frequency of infection-related explant surgeries.

Salmonella Typhi: A Review of Antibiogram Journey in Developing Countries.

BACKGROUND: Typhoid fever poses a significant health challenge in low- and middleincome countries (LMiCs), impacting millions of individuals across various age groups. Its prevalence is particularly pronounced in South Asia. Factors contributing to its transmission in South Asia include rapid unplanned urbanization, urban-rural disparities, provision of poor water and sanitation facilities, and open defecation. The mortality rate of typhoid fever is up to 1%, and those who survive have a protracted period of poor health and carry an enormous financial burden. The treatment is further complicated by the emerging antibiotic resistance leaving few treatment options in hands. This issue has become more urgent due to the further emergence of extended drug-resistant (XDR) and multidrugresistant (MDR) typhoid strains, as well as their subsequent global spread. Fluoroquinoloneresistant Salmonella spp. is currently classified by the World Health Organization (WHO) as a high (Priority 2) pathogen. As a result, establishing minimum inhibitory concentrations (MIC) according to the latest guidelines may prove effective in treating typhoid fever and minimizing the rising threat of drug resistance.

Ratiometric Fluorescent Probe Promotes Trans-differentiation of Human Mesenchymal Stem Cells to Neurons.

Development of multifunctional theranostics is challenging and crucial for deciphering complex biological phenomena and subsequently treating critical disease. In particular, development of theranostics for traumatic brain injury (TBI) and understanding its repair mechanism are challenging and highly complex areas of research. Recently, there have been interesting pieces of research work demonstrated that a small molecule-based neuroregenerative approach using stem cells has potential for future therapeutic lead development for TBI. However, these works demonstrated the application of a mixture of multiple molecules as a "chemical cocktail", which may have serious toxic effects in the differentiated cells. Therefore, development of a single-molecule-based potential differentiating agent for human mesenchymal stem cells (hMSCs) into functional neurons is vital for the upcoming neuro-regenerative therapeutics. This lead could be further extraploted for the design of theranostics for TBI. In this study, we have developed a multifunctional single-molecule-based fluorescent probe, which can image the transdifferentiated neurons as well as promote the differentiation process. We demonstrated a promising class of fluorescent probes (CP-4) that can be employed to convert hMSCs into neurons in the presence of fibroblast growth factor (FGF). This fluorescent probe was used in cellular imaging as its fluorescence intensity remained unaltered for up to 7 days of trans-differentiation. We envision that this imaging probe can have an important application in the study of neuropathological and neurodegenerative studies.

Emergence of Monkeypox (MPX): A Close Relative of Small Pox During COVID-19 Era.

After the eradication of smallpox (SPX), a new zoonotic threat that can trigger outbreaks has emerged. It may be fatal during the COVID19 outbreak. Humanity continues to be threatened due to re-emergence of the outbreaks. In most cases, new emerging viral agents originate from nonhuman hosts with zoonotic origins. Recent outbreaks of zoonotic infectious diseases with the potential to cause epidemics and pandemics continue to pose a major threat to the health security of entire regions, continents, and the world at large. Around five decades backthat Monkeypox (MPX) was reported for the first time in the Democratic Republic of the Congo (DRC) and was then confined to Central Africa only. Over the time, it has spread to other regions of Africa as well as outside Africa. As of August 2022, 40398 infections have been confirmed in almost 68 countries that have never reported MPX before. The majority of infections have been reported in Europe and Southeast Asia. On 23rd August 2022, MPX was declared a public health emergency of international concern, a step below declaring any disease as a pandemic. The article discusses the recent history of MPX outbreaks, as well as the evolving clinical manifestations of the disease, and the possible causes of the increase in cases, including the cessation of SPX vaccinations.

Risk factors, mortality, and predictors of survival in COVID-19-associated pulmonary mucormycosis: a multicentre retrospective study from India.

OBJECTIVES: To compare COVID-19-associated pulmonary mucormycosis (CAPM) with COVID-19-associated rhino-orbital mucormycosis (CAROM), ascertain factors associated with CAPM among patients with COVID-19, and identify factors associated with 12-week mortality in CAPM. METHODS: We performed a retrospective multicentre cohort study. All study participants had COVID-19. We enrolled CAPM, CAROM, and COVID-19 subjects without mucormycosis (controls; age-matched). We collected information on demography, predisposing factors, and details of COVID-19 illness. Univariable analysis was used to compare CAPM and CAROM. We used multivariable logistic regression to evaluate factors associated with CAPM (with hypoxemia during COVID-19 as the primary exposure) and at 12-week mortality. RESULTS: We included 1724 cases (CAPM [n = 122], CAROM [n = 1602]) and 3911 controls. Male sex, renal transplantation, multimorbidity, neutrophil-lymphocyte ratio, intensive care admission, and cumulative glucocorticoid dose for COVID-19 were significantly higher in CAPM than in CAROM. On multivariable analysis, COVID-19-related hypoxemia (aOR, 2.384; 95% CI, 1.209-4.700), male sex, rural residence, diabetes mellitus, serum C-reactive protein, glucocorticoid, and zinc use during COVID-19 were independently associated with CAPM. CAPM reported a higher 12-week mortality than CAROM (56 of the 107 [52.3%] vs. 413 of the 1356 [30.5%]; p = 0.0001). Hypoxemia during COVID-19 (aOR [95% CI], 3.70 [1.34-10.25]) and Aspergillus co-infection (aOR [95% CI], 5.40 [1.23-23.64]) were independently associated with mortality in CAPM, whereas surgery was associated with better survival. DISCUSSION: CAPM is a distinct entity with a higher mortality than CAROM. Hypoxemia during COVID-19 illness is associated with CAPM. COVID-19 hypoxemia and Aspergillus co-infection were associated with higher mortality in CAPM.

Effectiveness of prophylactic oral and/or vaginal probiotic supplementation in the prevention of recurrent urinary tract infections: A randomized, double-blind, placebo-controlled trial.

BACKGROUND: Widespread antibiotic resistance has sparked interest in the lookout for non-antibiotic strategies, particularly focusing on probiotics for the prevention of recurrent urinary tract infections (UTIs). We evaluated the effectiveness of prophylactic probiotic supplementation through oral and intravaginal routes in the prevention of recurrent UTIs. METHODS: This double-blind, placebo-controlled study enrolled 174 premenopausal women with a history of recurrent UTIs and randomized them to either of the four treatment groups, namely, Placebo (G1, oral placebo+vaginal placebo); Oral probiotic (G2, oral lactic acid bacteria and bifidobacteria+vaginal placebo); Vaginal probiotic (G3, oral placebo+vaginal lactobacilli); and Probiotic combination (oral lactic acid bacteria and bifidobacteria+vaginal lactobacilli), for 4 months. Participants were followed-up for symptomatic UTIs for one year. The primary endpoints were the number of symptomatic UTIs at 4 months, the proportion of subjects with at least one symptomatic UTI, and the time to the first symptomatic UTI. RESULTS: The incidence of UTI at 4 months in G1, G2, G3 and G4 was 70.4%, 61.3%, 40.9%, and 31.8%, respectively. The mean number of symptomatic UTI recurrence at 4 months was significantly lower (p<0.05) in G3 (1.06) and G4 (1.07) compared to G1 (2.1) and G2 (1.63). Further, the time to first symptomatic UTI (days) was significantly longer (p<0.05) in G3 (123.8) and G4 (141.8) compared to G1 (69.3) and G2 (71.9). Probiotic supplementations were well tolerated with no serious adverse events. CONCLUSION: Prophylactic supplementation with either vaginal probiotics or in combination with oral probiotics demonstrated effectiveness in preventing recurrent symptomatic UTI episodes.

Unsupervised machine learning to investigate trajectory patterns of COVID-19 symptoms and physical activity measured via the MyHeart Counts App and smart devices.

Previous studies have associated COVID-19 symptoms severity with levels of physical activity. We therefore investigated longitudinal trajectories of COVID-19 symptoms in a cohort of healthcare workers (HCWs) with non-hospitalised COVID-19 and their real-world physical activity. 121 HCWs with a history of COVID-19 infection who had symptoms monitored through at least two research clinic visits, and via smartphone were examined. HCWs with a compatible smartphone were provided with an Apple Watch Series 4 and were asked to install the MyHeart Counts Study App to collect COVID-19 symptom data and multiple physical activity parameters. Unsupervised classification analysis of symptoms identified two trajectory patterns of long and short symptom duration. The prevalence for longitudinal persistence of any COVID-19 symptom was 36% with fatigue and loss of smell being the two most prevalent individual symptom trajectories (24.8% and 21.5%, respectively). 8 physical activity features obtained via the MyHeart Counts App identified two groups of trajectories for high and low activity. Of these 8 parameters only 'distance moved walking or running' was associated with COVID-19 symptom trajectories. We report a high prevalence of long-term symptoms of COVID-19 in a non-hospitalised cohort of HCWs, a method to identify physical activity trends, and investigate their association. These data highlight the importance of tracking symptoms from onset to recovery even in non-hospitalised COVID-19 individuals. The increasing ease in collecting real-world physical activity data non-invasively from wearable devices provides opportunity to investigate the association of physical activity to symptoms of COVID-19 and other cardio-respiratory diseases.

The new Xpert Mycobacterium tuberculosis/rifampicin (MTB/Rif) Ultra assay in comparison to Xpert MTB/Rif assay for diagnosis of tuberculosis in children and adolescents.

BACKGROUND: Microbiological diagnosis of pediatric tuberculosis (TB) using conventional microbiological techniques has been challenging due to paucibacillary nature of the disease. Molecular methods using cartridge-based tests like Xpert, have immensely improved diagnosis. A novel next-generation cartridge test, Xpert Ultra, incorporates two additional molecular targets and claims to have much lower detection limit. We attempted to compare the two techniques in presumptive pediatric TB patients. OBJECTIVES: The aim of this study was to compare the diagnostic performance of Xpert MTB/Rif Ultra with Xpert MTB/Rif for the detection of pediatric TB. STUDY DESIGN: This is an observational comparative analytical study. METHODS: Children under 15 years of age with presumptive TB were enrolled. Appropriate specimens were obtained (sputum, induced sputum or gastric aspirate for suspected pulmonary TB, cerebrospinal fluid for suspected tubercular meningitis and pleural fluid for suspected tubercular pleural effusion), subjected to smear microscopy, mycobacterial culture, Xpert and Xpert ultra tests and other appropriate diagnostic investigations. RESULTS: Out of 130 enrolled patients, 70 were diagnosed with TB using a composite reference standard (CRS). The overall sensitivity of Xpert was 64.29% [95% confidence interval (CI) 51.93-75.93%] and that of Xpert Ultra was 80% (95% CI 68.73-88.61%) with 100% overall specificity for both. The sensitivity of Xpert and Xpert Ultra in pulmonary specimens (n = 112) was 66.67% and 79.37% and in extrapulmonary specimens (n = 18) was 42.86% and 85.71%, respectively. CONCLUSION: Our study found Ultra to be more sensitive than Xpert for the detection of Mycobacterium tuberculosis in children. Our findings support the use of Xpert Ultra as initial rapid molecular diagnostic test in children under evaluation for TB.

Triple Burden: The Incorrigible Threat of Tuberculosis, HIV, and COVID-19.

The Coronavirus-19 (COVID-19) hasn't seen the dawn since its emergence, however waxing and waning has resulted in the emergence of deadly variants. The effects of the pandemic have not been limited to its virulence, but have rather conferred multiple collateral effects, especially in developing countries; thereby, designating it as a SYNDEMIC. The same culminated in neglect of non-COVID-19 conditions like tuberculosis (TB) and human immunodeficiency virus-acquired immunodeficiency syndrome (HIV/AIDS). Besides being the prognostic factor for severe COVID-19, these infections in hidden pockets served as a reservoir for the emergence of the deadly Omicron. Another significant impact of this juxtaposition was on the delivery of healthcare services for TB and HIV. The unanticipated COVID-19 pandemic turned the path of ongoing progress of elimination programs. Direct consequences of the COVID-19 pandemic were pronounced on diagnosis, treatment, and services for patients with TB and HIV. Essential TB services were reallocated to the COVID-19 rapid response task force. However, despite escalating the tribulations, this triple burden has simultaneously taught lessons to escalate the progress of halted programs. The pandemic has catalyzed an unusual level of collaboration among scientists, which can be exploited for TB and HIV. Fast-track diagnostics, digitalization, contact tracing, and vaccine development have enabled the world to envision the same for TB/HIV.

Dual immunization with CdtB protein and flagellin epitope offers augmented protection against enteric fever in mice.

AIMS: Present study has explored the protective response of dual immunization using two different antigenic entities (i.e. flagellin epitope and cytolethal distending toxin subunit B (CdtB) protein) against lethal challenge of typhoidal serovars in a murine model. MAIN METHODS: In-vitro immunogenicity of flagellin epitope-BSA conjugate and CdtB protein was confirmed using Indirect ELISA of typhoid positive patients' sera. Further, both entities were administered intraperitoneally in mice individually or in combination, followed by lethal challenge of typhoidal Salmonellae. Various parameters were analysed such as bacterial burden, mice survival, histopathological analysis, cytokine analysis and immunophenotyping. Serum samples obtained from the immunized mice were used for passive immunization studies, wherein mice survival and mechanism of action of the generated antibodies was studied. KEY FINDINGS: Active immunization studies using the combination of both entities demonstrated improved mice survival after lethal challenge with typhoidal Salmonellae, reduced bacterial burden in organs, expression of immunophenotypic markers in splenocytes and restored tissue histoarchitecture. When used in combination, the effective doses of both the candidates reduced which may be attributed to multiprong approach used by the immune system to recognize Salmonella. Passive immunization studies further determined the protective efficacy of generated antibodies by different mechanisms such as complement mediated bactericidal action, swarming inhibition and increased phagocytic uptake. SIGNIFICANCE: Present study is the first phase of the proof-of-concept which may prove to be beneficial in developing an effective bi-functional vaccine candidate to render protection against both Vi-positive as well as Vi-negative Salmonella strains.

A Double Hit to Ubiquitination Leading to a New Diagnosis of VEXAS Syndrome.

VEXAS (vacuoles, E1 enzyme, X-linked, auto-inflammatory, somatic) syndrome is a newly defined illness that bridges hematology, oncology, and rheumatology. Its pathophysiology originates in a mutation in the UBA1 gene that leads to a defect in ubiquitination resulting in a severe systemic inflammatory syndrome. It is associated with significant morbidity and mortality; however, data are scarce due to limited cases described in the literature. Here we describe a case of a male in his 60s who was referred to hematology-oncology due to progressive dyspnea, poor oral intake, and weight loss. He was diagnosed with relapsing polychondritis 2 years prior; however, his symptoms did not improve despite treatment. He was ultimately diagnosed with VEXAS syndrome with a mutation in UBA1 (ubiquitin-like modifier activating enzyme 1) and a concurrent SQSTM1 mutation. In addition, the coexistence of two mutations in the ubiquitination pathway in the same patient has not been reported to date. This patient and the treatment course were compared to pre-existing literature to increase awareness and improve the medical management of VEXAS syndrome.

Risk factor and resistance profile of colistin resistant Acinetobacter baumannii and Klebsiellapneumoniae.

PURPOSE: Antimicrobial resistance is one of the major global health concerns, which is relentless despite multipronged measures. Carbapenems and colistin, drug of choice for multi drug resistant Klebsiella pneumoniae and Acinetobacter species, have also been rendered of less use. This underlines the need to decipher prevalence of colistin resistance comprehensively for formulation of hospital and country-wise antibiogram. We conducted this study to decipher the prevalence of colistin resistance in our tertiary care centre of North India. MATERIALS AND METHODS: This was a prospective, case control study conducted over a period of one and half years. All carbapenem resistant Klebsiella pneumoniae and Acinetobacter isolates were included. Kirby-Bauer method of disc diffusion was used for all antibiotics, except colistin for which broth microdilution was performed and interpreted using CLSI guidelines. Demographic details, risk factors and outcome details were recorded. Genotypic characterization was performed using representative strains, for blaNDM, blaKPC and blaOXA-48. RESULTS: Of 103 carbapenem resistant isolates, 7 were found to be colistin resistant. Median age was 43 years, with male:female ratio of 1.1:1. 35% isolates were from pus samples, followed by endotracheal aspirate. Colistin resistance was more in ICUs than wards. Presence of indwelling devices was noted as the most common risk factor, followed by previous antibiotic exposure and use of steroids/immunosuppressants. Indwelling devices, steroids/immunosuppressants usage, length of hospital stay, COPD, prior usage of carbapenems, piperacillin-tazobactam and colistin, usage of ampicillin-sulbactam during hospital stay, were statistically significant. Mortality was noted in 4 cases, with statistical difference between control and case arm. The blaNDM and blaOXA-48 were noted in 3 and 2 isolates respectively, with absence of blaKPC. CONCLUSION: The present study unravels incidence, risk factors and resistance encoding genes at our centre. This is of immense help in formulation of antibiotic policies and guidance for infection control measures.

A rare case of genital myiasis in a patient with genital prolapse.

Genital myiasis is an infestation of genital organs by fly larvae, where they feed and develop as parasites. They can cause severe infection, inflammatory reaction and can be linked to psychiatric disturbances. We report a rare case of genital myiasis in an elderly postmenopausal woman aged 82 years from Udaipur, Rajasthan. She presented with complaints of intense pain in the genital region and was diagnosed as a case of genital myiasis of a prolapsed uterus. Pelvic examination revealed 'Stage-IV' genitourinary prolapse according to Pelvic Organ Prolapse-Quantification (POP-Q classification), with a big excavatory ulcer indwelled with maggots of Musca domestica. About 100 such maggots were subsequently manually removed with forceps. With proper surgical and nonsurgical interventions, the patient healed completely and the prolapse was reduced completely.

Emergence of COVID-19 Variants: An Update.

Severe acute respiratory disease virus-2 (SARS CoV-2) is one of the deadliest global threats faced by mankind to date. Despite the colossal efforts, the viral pandemic swept across all boundaries. Besides the virulence and susceptible population, the low proofreading capacity and error-prone mechanism of RNA-dependent RNA polymerase (RdRp) have contributed to new variants and reinfections. The World Health Organization has officially categorized these variants as variants of concern or variants of interest. This nomenclature is not merely to suffice the surveillance but also to have effective treatment and vaccine options in place. Coronavirus disease 2019 (COVID-19) variants have the propensity to render the available treatment strategies futile owing to the mutations they acquire. The futility of treatment strategies can be attributed either to the ineffectiveness or the shortage of supply given the skyrocketing increase in the number of cases. Presently, the Omicron variant is the most widespread one and is known to escape the protection, be it immune-derived, vaccination-derived, or hybrid. WHO has recommended modification in vaccine development policies and few companies have introduced Omicron-adapted vaccine jabs. Keeping in view the unending tale of COVID-19 variants and the huge data available on the same, this review focuses on providing insight into the emergence and ongoing dynamics of these new COVID-19 variants.

Primary Cutaneous Mucormycosis: A Necrotising Soft Tissue Infection with Poor Prognosis.

BACKGROUND: Cutaneous mucormycosis is an unusual fungal infection that continues to occur. It needs aggressive surgical debridement and timely administration of antifungals due to its high fatality rate. High clinical suspicion on the part of a surgeon is required to prevent the same. CASE PRESENTATION: We present two cases of cutaneous mucormycosis in which the patients succumbed to death, highlighting the seriousness of the condition. One patient had a lower leg ulcer and was diabetic, and the other patient had a gluteal abscess following an intramuscular injection. Tissue samples grew Rhizopus arrhizus and Apophysomyces sp., respectively. Both patients were treated with amphotericin B, and extensive debridement was performed. DISCUSSION: Cutaneous mucormycosis can be reported in immunocompetent people, and there is a need for early recognition of the entity as a differential diagnosis of any nonhealing necrotic ulcer. CONCLUSION: Proper training and education of technical and clinical staff should be done at peripheral primary and secondary care centres so as not to miss out on cases of mucormycosis and for better prognosis in a cutaneous variety of mucormycosis in surgical patients.

Controlling Amyloid Beta Peptide Aggregation and Toxicity by Protease-Stable Ligands.

Polymerization of soluble amyloid beta (Aß) peptide into protease-stable insoluble fibrillary aggregates is a critical step in the pathogenesis of Alzheimer's disease (AD). The N-terminal (NT) hydrophobic central domain fragment 16KLVFF20 plays an important role in the formation and stabilization of ß-sheets by self-recognition of the parent Aß peptide, followed by aggregation of Aß in the AD brain. Here, we analyze the effect of the NT region inducing ß-sheet formation in the Aß peptide by a single amino acid mutation in the native Aß peptide fragment. We designed 14 hydrophobic peptides (NT-01 to NT-14) by a single mutation at 18Val by using hydrophobic residues leucine and proline in the natural Aß peptide fragment (KLVFFAE) and analyzed its effect on the formation of Aß aggregates. Among all these peptides, NT-02, NT-03, and NT-13 significantly affected the Aß aggregate formation. When the NT peptides were coincubated with the Aß peptide, a significant reduction in ß-sheet formation and increment in random coil content of Aß was seen, confirmed by circular dichroism spectroscopy and Fourier transform infrared spectroscopy, followed by the reduction of fibril formation measured by the thioflavin-T (ThT) binding assay. The aggregation inhibition was monitored by Congo red and ThT staining and electron microscopic examination. Moreover, the NT peptides protect the PC-12 differentiated neurons from Aß-induced toxicity and apoptosis in vitro. Thus, manipulation of the Aß secondary structure with protease-stable ligands that promote the random coil conformation may provide a tool to control the Aß aggregates observed in AD patients.