The NLRP3 Inflammasome Is Dispensable in Methicillin-Resistant Staphylococcus aureus Urinary Tract Infection.

The NLRP3 inflammasome is a cytoplasmic complex that senses molecular patterns from pathogens or damaged cells to trigger an innate immune defense response marked by the production of proinflammatory cytokines IL-1ß and IL-18 and an inflammatory death called pyroptosis. The NLRP3 inflammasome is activated in the urinary tract by a variety of infectious and non-infectious insults. In this study, we investigated the role of the NLRP3 inflammasome by comparing the pathophysiology of methicillin-resistant Staphylococcus aureus (MRSA) ascending UTI in wild-type (WT) and Nlrp3-/- mice. The difference in the bacterial burden detected in the urinary tracts of MRSA-infected WT and Nlrp3-/- was not statistically significant at 6, 24, and 72 h post-infection (hpi). The levels of pro-inflammatory cytokines and chemokines as well as the numbers of granulocytes recruited to bladder and kidney tissues at 24 hpi were also similar between Nlrp3-/- and WT mice. The histopathological analysis of MRSA-infected bladder and kidney sections from Nlrp3-/- and WT mice showed similar inflammation. Overall, these results suggest that MRSA-induced urinary NLRP3 activity does not play a role in the pathophysiology of the ascending UTI.

Defining the Roles of Pyruvate Oxidation, TCA Cycle, and Mannitol Metabolism in Methicillin-Resistant Staphylococcus aureus Catheter-Associated Urinary Tract Infection.

Methicillin-resistant Staphylococcus aureus (MRSA) is an important cause of complicated urinary tract infection (UTI) associated with the use of indwelling urinary catheters. Previous reports have revealed host and pathogen effectors critical for MRSA uropathogenesis. Here, we sought to determine the significance of specific metabolic pathways during MRSA UTI. First, we identified four mutants from the Nebraska transposon mutant library in the MRSA JE2 background that grew normally in rich medium but displayed significantly reduced growth in pooled human urine (HU). This prompted us to transduce the uropathogenic MRSA 1369 strain with the transposon mutants in sucD and fumC (tricarboxylic acid [TCA] cycle), mtlD (mannitol metabolism), and lpdA (pyruvate oxidation). Notably, sucD, fumC, and mtlD were also significantly upregulated in the MRSA 1369 strain upon exposure to HU. Compared to the WT, the MRSA 1369 lpdA mutant was significantly defective for (i) growth in HU, and (ii) colonization of the urinary tract and dissemination to the kidneys and the spleen in the mouse model of catheter-associated UTI (CAUTI), which may be attributed to its increased membrane hydrophobicity and higher susceptibility to killing by human blood. In contrast to their counterparts in the JE2 background, the sucD, fumC, and mtlD mutants in the MRSA 1369 background grew normally in HU; however, they displayed significant fitness defects in the CAUTI mouse model. Overall, identification of novel metabolic pathways important for the urinary fitness and survival of MRSA can be used for the development of novel therapeutics. IMPORTANCE While Staphylococcus aureus has historically not been considered a uropathogen, S. aureus urinary tract infection (UTI) is clinically significant in certain patient populations, including those with chronic indwelling urinary catheters. Moreover, most S. aureus strains causing catheter-associated UTI (CAUTI) are methicillin-resistant S. aureus (MRSA). MRSA is difficult to treat due to limited treatment options and the potential to deteriorate into life-threatening bacteremia, urosepsis, and shock. In this study, we found that pathways involved in pyruvate oxidation, TCA cycle, and mannitol metabolism are important for MRSA fitness and survival in the urinary tract. Improved understanding of the metabolic needs of MRSA in the urinary tract may help us develop novel inhibitors of MRSA metabolism that can be used to treat MRSA-CAUTI more effectively.

Acute Gastroenteritis Induced Adrenal Crisis in a patient with Congenital Adrenal Hyperplasia: A Case Report.

Congenital Adrenal Hyperplasia is a group of autosomal recessive diseases due to deficiencies of enzymes involved in steroidogenesis. If not diagnosed and treated adequately, Congenital Adrenal Hyperplasia can lead to an acute adrenal crisis with hemodynamic collapse. Acute stressors and steroid insufficiency precipitate an adrenal crisis. The major clinical features are hypotension and volume depletion. Nonspecific symptoms such as fatigue, lack of energy, anorexia, nausea, vomiting, and abdominal pain are common. We report a case of a 3-year male previously diagnosed with congenital adrenal hyperplasia in adrenal crisis secondary to medicine noncompliance and gastroenteritis. The diagnosis was based on the clinical history and biochemical investigations. After initial resuscitation was managed; lifelong oral prednisolone and fludrocortisone were prescribed. Keywords: adrenal insufficiency; gastroenteritis; glucocorticoids.

Emergency unit assessment of seven tertiary hospitals in Nepal using the WHO tool: a cross-sectional study.

BACKGROUND: In 2021, the Nepal national emergency care system's assessment (ECSA) identified 39 activities and 11 facility-specific goals to improve care. To support implementation of the ECSA facility-based goals, this pilot study used the World Health Organization's (WHO) Hospital Emergency Unit Assessment Tool (HEAT) to evaluate key functions of emergency care at tertiary hospitals in Kathmandu, Nepal. METHODS: This cross-sectional study used the standardized HEAT assessment tool. Data on facility characteristics, human resources, clinical services, and signal functions were gathered via key informant interviews conducted by trained study personnel. Seven tertiary referral centers in the Kathmandu valley were selected for pilot evaluation including governmental, academic, and private hospitals. Descriptive statistics were generated, and comparative analyses were conducted. RESULTS: All facilities had continuous emergency care services but differed in the extent of availability of each item surveyed. Academic institutions had the highest rating with greater availability of consulting services and capacity to perform specific signal functions including breathing interventions and sepsis care. Private institutions had the highest infrastructure availability and diagnostic testing capacity. Across all facilities, common barriers included lack of training of key emergency procedures, written protocols, point-of-care testing, and ancillary patient services. CONCLUSION: This pilot assessment demonstrates that the current emergency care capacity at representative tertiary referral hospitals in Kathmandu, Nepal is variable with some consistent barriers which preclude meeting the ECSA goals. The results can be used to inform emergency care development within Nepal and demonstrate that the WHO HEAT assessment is feasible and may be instructive in systematically advancing emergency care delivery at the national level if implemented more broadly.

Correlation between neurologic status and spinal injury at the Cervicothoracic Junction.

BACKGROUND: With limited studies on spinal injuries occurring at the cervicothoracic junction, there is currently a knowledge gap regarding the correlation between morphology of injury and neurology and whether surgery provides a favorable neurological outcome. The primary objective was to determine whether the neurological deficit correlated with the severity of injury at this region of the spine. METHODS: All patients with injuries at the cervicothoracic junction from December 2015 to December 2020 in a government trauma hospital were included. Patient demographics, characteristics of the injury, neurological score, imaging findings, surgery details and neurological outcomes were analyzed. All patients had a minimum follow up of 2 years. RESULTS: Of the total 30 patients, 23 were male and 7 female with mean age 42.4 years. 90% had fall injuries with 76.7% sustaining AO type C injury and 10% with AO B2 injury.73.4% had injury at C6-C7 level followed by 13.3% , C7-T1. Only 16.7% patients presented with intact neurology. Plain x-rays failed to detect cervicothoracic junction, injuries in 63.3% patients. Posterior stabilization was performed in 56.7%. Neurological improvement was observed in 9 patients. CONCLUSIONS: Though cervicothoracic junction injuries are uncommon, they are highly unstable injuries and difficult to diagnose by plain x-rays. These injuries also result in profound neurological deficit. Surgical stabilization of these injuries should be considered for a favorable neurological and functional outcome.

Systematic Review of Literature Examining Bacterial Urinary Tract Infections in Diabetes.

This systematic review addresses the central research question, "what is known from the published, peer-reviewed literature about the impact of diabetes on the risk of bacterial urinary tract infections (UTI)?" We examine the results from laboratory studies where researchers have successfully adapted mouse models of diabetes to study the pathophysiology of ascending UTI. These studies have identified molecular and cellular effectors shaping immune defenses against infection of the diabetic urinary tract. In addition, we present evidence from clinical studies that in addition to diabetes, female gender, increased age, and diabetes-associated hyperglycemia, glycosuria, and immune impairment are important risk factors which further increase the risk of UTI in diabetic individuals. Clinical studies also show that the uropathogenic genera causing UTI are largely similar between diabetic and nondiabetic individuals, although diabetes significantly increases risk of UTI by drug-resistant uropathogenic bacteria.

Reliability and Validity of the Adapted Nepali Version of the AO Spine Patient Reported Outcome Spine Trauma.

BACKGROUND: The AO Spine Patient Reported Outcome Spine Trauma has been validated in English and Dutch language, however, there is an absence of a translated and validated version in Nepali language. The purpose of this study was to translate the AO Spine Patient Reported Outcome Spine Trauma into Nepali and adapt cross-culturally as outlined by established guidelines, as well as test its psychometric properties among Nepali speaking spine trauma patients. METHODS: Patients were recruited from two Nepali centers as a cross-sectional multicenter validation study. The English version of AO Spine Patient Reported Outcome Spine Trauma was translated and cross-culturally adapted into Nepali language following international guidelines. Next to AO Spine Patient Reported Outcome Spine Trauma also the EQ-5D-3L was filled out by the patients for concurrent validity. Descriptive statistics were used to analyze the patient characteristics. Assessment of measurement properties included content validity (floor and ceiling effects), internal consistency (Cronbach's ? and item total-correlation coefficients) and test-retest reliability by the Bland-Altman plot and Intraclass Correlation Coefficients. Spearman correlation tests were performed within the items and in correlation to EQ-5D-3L. RESULTS: Sixty two spine trauma patients completed the instrument with a mean time of 6.8 minutes. The translated version showed good content validity with no floor and ceiling effects. The internal consistency was excellent with a Cronbach's ? of 0.95. The Spearman correlations within the AO Spine Patient Reported Outcome Spine Trauma items were 0.07 - 0.65 and the test-retest analysis showed excellent results with an Intraclass Correlation Coefficients value of 0.95 (CI 0.93 - 0.97). Inverse correlation was observed between Nepali AO Spine PROST with EQ-5D-3L components. CONCLUSIONS: The Nepali version of AO Spine Patient Reported Outcome Spine Trauma demonstrated excellent validity and reliability results for measuring patient-reported outcomes of spine trauma patients.

Stromal Keratitis among Herpes Simplex Keratitis Patients in a Tertiary Eye Hospital: A Descriptive Cross-sectional Study.

Introduction: The manifestations of herpes simplex virus keratitis range from epithelial keratitis to vision-threatening stromal keratitis. There are limited studies done on our part regarding stromal keratitis. The aim of the study was to find out the prevalence of stromal keratitis among herpes simples keratitis patients in a tertiary eye hospital. Methods: This descriptive cross-sectional study was conducted among patients who presented with herpes simplex virus keratitis in a tertiary eye hospital between 1 January 2020 to 28 February 2022. Ethical clearance was taken from Ethical Review Board (Reference number: 1/2079/80). Data was collected from hospital records which was reviewed and visual acuity at presentation and at one-month follow-up, clinical details on examination were recorded and, diagnosis of the stage of disease given in the case record was noted. Convenience sampling was used. Point estimate and 95% Confidence Interval were calculated. Results: Among 112 Herpes simplex keratitis patients, the prevalence of stromal keratitis was 38 (33.93%) (25.16-42.70, 95% Confidence Interval). Conclusions: The prevalence of stromal keratitis among patients of herpes simplex keratitis was similar to studies conducted in similar settings. Keywords: herpes simplex keratitis; keratitis; prevalence.

Human Urine Alters Methicillin-Resistant Staphylococcus aureus Virulence and Transcriptome.

Gram-positive methicillin-resistant Staphylococcus aureus (MRSA) is an emerging cause of hospital-associated urinary tract infections (UTI), especially in catheterized individuals. Despite being rare, MRSA UTI are prone to potentially life-threatening exacerbations such as bacteremia that can be refractory to routine antibiotic therapy. To delineate the molecular mechanisms governing MRSA urinary pathogenesis, we exposed three S. aureus clinical isolates, including two MRSA strains, to human urine for 2 h and analyzed virulence characteristics and changes in gene expression. The in vitro virulence assays showed that human urine rapidly alters adherence to human bladder epithelial cells and fibronectin, hemolysis of sheep red blood cells (RBCs), and surface hydrophobicity in a staphylococcal strain-specific manner. In addition, transcriptome sequencing (RNA-Seq) analysis of uropathogenic strain MRSA-1369 revealed that 2-h-long exposure to human urine alters MRSA transcriptome by modifying expression of genes encoding enzymes catalyzing metabolic pathways, virulence factors, and transcriptional regulators. In summary, our results provide important insights into how human urine specifically and rapidly alters MRSA physiology and facilitates MRSA survival in the nutrient-limiting and hostile urinary microenvironment. IMPORTANCE Methicillin-resistant Staphylococcus aureus (MRSA) is an uncommon cause of urinary tract infections (UTI) in the general population. However, it is important to understand MRSA pathophysiology in the urinary tract because isolation of MRSA in urine samples often precedes potentially life-threatening MRSA bacteremia. In this report, we describe how exposure to human urine alters MRSA global gene expression and virulence. We hypothesize that these alterations may aid MRSA in acclimating to the nutrient-limiting, immunologically hostile conditions within the urinary tract leading to MRSA UTI.

May Measurement Month 2019: an analysis of blood pressure screening results from Nepal.

May Measurement Month (MMM) is a global initiative to screen high blood pressure (BP) in the community and increase awareness at the population level. High BP is the leading risk factor for mortality worldwide and in Nepal. This study presents the results of the 2019 MMM in Nepal. Opportunistic BP screening was conducted in 30 out of 77 districts across Nepal and aged ≥18 years at the community and public places. BP was measured three times in a seated position. A total of 74 205 individuals participated in the study, mean age 39.9 years, and 58% were male. BP measurements for the second and third readings were available for 69 292 (93.3%) individuals. The proportion of the population that were hypertensive was 27.5% (n = 20 429). Among those hypertensives, 46.3% were aware of their hypertensive status and of these, 37.5% were on antihypertensive medication. Only 54.3% of those on antihypertensive medication had their BP controlled. Of the community screened, those self reporting to have diabetes, current tobacco users, and current alcohol drinkers were 6.7%, 23.6%, and 31.9%, respectively; 20.6% of the participants were overweight, and 6.5% were obese. Since the first BP screening campaign, MMM 2017 in Nepal, the number of participants screened has largely increased over the years. MMM's success in Nepal is through a coordinated mobilization of trained health science students and volunteers in the communities. The Nepal MMM data demonstrates that large community-based BP screening campaigns are possible in low resource settings.

Epidemiology of Trauma Patients Presented at Emergency Department of Trauma Center.

BACKGROUND: Emergency Department of National Trauma Center Nepal Center is the only specialized trauma care hospital in Nepal, in operation since 2012. Traumatic injury is one of the major causes for mortality worldwide. This study aims to see the epidemiology, pattern of injuries and outcome of the patients presenting to the emergency room. METHODS: After getting ethical approval, we included all patients presenting to the Emergency Department with at least one injury, between Jan 2018 to Dec 2020. Informations on age, gender, mechanism of injury and outcome was abstracted for all patients presenting to the emergency department. Data were extracted from hospital database with the permission of hospital authority. RESULTS: Total of 49991 patients presented to emergency department with different types of injuries.  Among them 7792 (14.0%) needed hospital admission. Fall and road crashes comprised almost 80% of admitted cases. Mortality was 2.1% of admitted patients, mostly with head injuries followed by multiple injuries. CONCLUSIONS: Among the patient visiting the trauma center during the study period most common mechanism of the injury were fall and road crash with head and neck injury. Majority of the patients were only managed in the ward without surgery.

Exposure to Moderate Glycosuria Induces Virulence of Group B Streptococcus.

To explore whether glycosuria induces virulence of uropathogens, in turn facilitating urinary tract infection (UTI), we exposed group B Streptococcus (GBS) strain 10/84 to human urine plain or with 300 mg/dL glucose (mimicking moderate glycosuria). Exposure to moderate glycosuria significantly augmented bacterial growth, kidney bacterial burden in a mouse model of ascending UTI, and virulence characteristics and expression of corresponding genes. Exposure to glycosuria increased GBS adherence to human bladder epithelial cell line and expression of corresponding PI2a fimbrial gene, antimicrobial peptide LL-37 resistance and bacterial surface charge modulating dltA, and GBS hemolytic ability and expression of genes encoding pore-forming toxins.

Electronic Cigarette (E-Cigarette) Vapor Exposure Alters the Streptococcus pneumoniae Transcriptome in a Nicotine-Dependent Manner without Affecting Pneumococcal Virulence.

The effects of electronic cigarette (e-cigarette) vapor (EV) exposure on the physiology of respiratory microflora are not fully defined. We analyzed the effects of exposure to vapor from nicotine-containing and nicotine-free e-liquid formulations on the virulence and transcriptome of Streptococcus pneumoniae strain TIGR4, a pathogen that asymptomatically colonizes the human nasopharyngeal mucosa. TIGR4 was preexposed for 2 h to nicotine-containing EV extract (EVE+NIC), nicotine-free EV extract (EVE-NIC), cigarette smoke extract (CSE), or nutrient-rich tryptic soy (TS) broth (control). The differences between the treatment and control strains were explored using transcriptome sequencing (RNA sequencing [RNA-Seq]), in vitro virulence assays, and an in vivo mouse model of acute pneumonia. The analysis of RNA-Seq profiles revealed modest changes in the expression of 14 genes involved in sugar transport and metabolism in EVE-NIC-preexposed TIGR4 compared to the control, while EVE+NIC or CSE exposure altered expression of 264 and 982 genes, respectively, most of which were involved in metabolism and stress response. Infection in a mouse model of acute pneumonia with control TIGR4 or with TIGR4 preexposed to EVE+NIC, EVE-NIC, or CSE did not show significant differences in disease parameters, such as bacterial organ burden and respiratory cytokine response. Interestingly, TIGR4 exposed to CSE or EVE+NIC (but not EVE-NIC) exhibited moderate induction of biofilm formation. However, none of the treatment groups showed significant alterations in pneumococcal hydrophobicity or epithelial cell adherence. In summary, our study reports that exposure to EV significantly alters the S. pneumoniae transcriptome in a nicotine-dependent manner without affecting pneumococcal virulence.IMPORTANCE With the increasing popularity of e-cigarettes among cigarette smoking and nonsmoking adults and children and the recent reports of vaping-related lung illness and deaths, further analysis of the adverse health effects of e-cigarette vapor (EV) exposure is warranted. Since pathogenic bacteria such as Streptococcus pneumoniae can colonize the human nasopharynx as commensals, they may be affected by exposure to bioactive chemicals in EV. Hence, in this study we examined the effects of EV exposure on the physiology of S. pneumoniae strain TIGR4. In order to differentiate between the effects of nicotine and nonnicotine components, we specifically compared the RNA-Seq profiles and virulence of TIGR4 exposed to vapor from nicotine-containing and nicotine-free e-liquid formulations. We observed that nicotine-containing EV augmented TIGR4 biofilms and altered expression of TIGR4 genes predominantly involved in metabolism and stress response. However, neither nicotine-containing nor nicotine-free EV affected TIGR4 virulence in a mouse model.

Epidemiological Characteristics of Dorsal and Lumbar Spine Trauma Presenting to a Trauma Hospital in Kathmandu, Nepal: Formulation of a National Spine Policy.

INTRODUCTION: Outcome of spine injury treated in resource constrained regions may not be the same as in developed nations. The aim of the present study was to study the epidemiological characteristics, delay, complications, and outcome of surgically treated dorsal and lumbar trauma. METHODS: Retrospective study of dorsal and lumbar spine injury patients treated between December 2015 and August 2017. Patients were segregated into four groups based on the timing of surgery: 0-2 days, 3-7 days, 8-30 days, and more than 31 days. Only one operating room twice a week was allotted to spine surgery, and spine had to compete with orthopedic and surgical trauma for admission and surgery. RESULTS: Ninety-one patients (male 61) with mean age 33 years were operated for dorsal and lumbar spine injuries. 84% of the total patients sustained a fall, and 86.8% were from the periphery. Though 69.2% presented within 2 days, only 4.4% were operated within 2 days. Majority of the delay was due to unavailability of the operating room followed by financial constraints. Twenty-seven patients had complete deficit, 32 incomplete deficit, and 32 normal neurology. Four patients operated within 2 days improved their neurology, 7 incomplete deficit patients in 3-7 days group improved, 6 in 8-30 days group improved, whereas no patient in more than 31 days group improved. Overall 53.1% of neurologically incomplete deficit patients improved if operated within 30 days. No neurological improvement was seen in the 27 complete deficit patients. Wound infection, pulmonary contusion, and deep vein thrombosis were seen in 3 patients. CONCLUSIONS: As expected 95.6% of our patients were treated more than 3 days after injury and 60% more than a week later, which may not be acceptable in advanced countries. Despite the delay, 53.1% had an improvement in neurology when operated within 30 days. Hence, surgery still holds the hope of neurological recovery and quicker rehabilitation.