Clinical, laboratory profile and molecular characterization of Orientia tsutsugamushi among fatal scrub typhus patients from Karnataka, India.

BACKGROUND: Scrub typhus is a vector-borne infection caused by the obligate intracellular organism Orientia tsutsugamushi. In some cases, scrub typhus can result in severe complications, multiorgan failure and death. OBJECTIVE: To study the clinical and laboratory profiles of patients who succumbed to scrub typhus. METHODS: A prospective cohort study was conducted from August 2019 through April 2023 on scrub typhus patients admitted to our hospital. Clinical and laboratory parameters of all the patients were recorded, and blood samples were drawn. To confirm scrub typhus, a nested polymerase chain reaction (nPCR) was performed in collected samples. Viable amplicons were sequenced, and phylogenetic analyses were performed to identify infecting genotypes. RESULTS: A total of 261 patients were enrolled. Of these, nine (3.45%) patients succumbed at a median (Interquartile Range) duration of 5 (1.5, 10.5) days after admission. Sepsis with septic shock (9, 100%) and acute kidney injury (AKI) (6, 66%) were noted among the succumbed patients. All the succumbed patients (100%) required intensive care admission, inotropic and ventilatory support. While 5 (55%) patients required dialysis, two (22%) required blood transfusion. Three (33%) patient samples were co-positive for Leptospira IgM, and four (44%) patients had superinfection with Candida tropicalis, multi-drug-resistant (MDR) E. Coli sepsis, pan drug-resistant (PDR) Acinetobacter Baumanii, and Klebsiella pneumoniae. Phylogenetic analysis revealed Orientia tsutsugamushi Japanese Gilliam-variant (JG-v) like (50%), Karp-like (37.5%), and Japanese Gilliam (JG) like (12.5%) strains among succumbed patients. CONCLUSION: Delay in scrub typhus diagnosis can result in severe complications, septic shock, and multisystem organ failure, culminating in death.

Clinical, laboratory, and molecular epidemiology of Orientia tsutsugamushi infection from Southwestern India.

Scrub typhus is a vector borne disease which in a proportion of patients causes multiorgan involvement and death if untreated. Infecting genotype and virulence factors play a role in severity of infection and outcome. The current prospective cohort study was undertaken to elucidate the severity of illness in scrub typhus patients and to identify the circulating genotypes in Karnataka, India. A total of 214 patients of either gender from 9 districts of Karnataka and one patient each from Andhra Pradesh and Kerala, India were enrolled in the study. With a predefined severity criterion, 132 patients were segregated to the severe group. Multi organ involvement was seen in 59 (44.69%) patients. Phylogenetic analysis revealed JG-v like (48.97%), Karp-like (26.53%), JG-like (22.44%), and Kato-like (2.04%) strains in Karnataka. Patients infected with Orientia tsutsugamushi Karp-like strains had respiratory involvement (69.2%), cardiovascular involvement (46.2%) and thrombocytopenia (23.1%) and required higher hospital resource utilization.

Comparison of Hemagglutination and Hemolytic Activity of Various Bacterial Clinical Isolates Against Different Human Blood Groups.

Among the various pathogenic determinants shown by microorganisms hemagglutination and hemolysin production assume greater significance in terms of laboratory identification. This study evaluated the hemagglutination and hemolytic activity of various bacterial isolates against different blood groups. One hundred and fifty bacterial strains, isolated from clinical specimens like urine, pus, blood, and other body fluids were tested for their hemagglutinating and hemolytic activity against human A, B, AB, and O group red blood cells. Among the 150 isolates 81 were Escherichia coli, 18 were Klebsiella pneumoniae, 19 were Pseudomonas aeruginosa, 10 were Pseudomonas spp, six were Proteus mirabilis, and the rest 16 were Staphylococcus aureus. Nearly 85% of the isolates agglutinated A group cells followed by B and AB group (59.3% and 60.6% respectively). Least number of isolates agglutinated O group cells (38.0%). When the hemolytic activity was tested, out of these 150 isolates 79 (52.6%) hemolyzed A group cells, 61 (40.6%) hemolyzed AB group cells, 46 (30.6%) hemolyzed B group cells, and 57 (38.6%) isolates hemolyzed O group cells. Forty-six percent of the isolates exhibited both hemagglutinating and hemolytic property against A group cells, followed by B and AB group cells (28.6% and 21.3% respectively). Least number of isolates i.e., 32 (21.3%) showed both the properties against O group cells. The isolates showed wide variation in their hemagglutination and hemolytic properties against different combinations of human blood group cells. The study highlights the importance of selection of the type of cells especially when human RBCs are used for studying the hemagglutination and hemolytic activity of bacterial isolates because these two properties are considered as characteristic of pathogenic strains.

Evaluation of Different Phenotypic Techniques for the Detection of Slime Produced by Bacteria Isolated from Clinical Specimens.

UNLABELLED: INTRODUCTION : Microorganisms use various strategies for their survival in both the environment and in humans. Slime production by bacteria is one such mechanism by which microbes colonize on the indwelling prosthetic devices and form biofilms. Infections caused by such microorganisms are difficult to treat as the biofilm acts as a shield and protects microbes against antimicrobial agents. There are several methods for the detection of slime produced by bacteria, and they include both phenotypic and molecular methods. The present study evaluated the Congo red agar/broth method, Christensen's method, dye elution technique, and the latex agglutination method for the demonstration of slime production by different bacterial clinical isolates. MATERIALS & METHODS: We collected 151 bacterial clinical isolates (both gram-positive and gram-negative bacteria) from various specimens and tested them for the production of slime both by qualitative and quantitative tests. Congo red agar/broth method, Christensen's method, dye elution technique, and latex agglutination methods were used for detecting the slime or slime-like substance. RESULTS : We found that 103 (68.2%) strains were positive for slime production by Congo red agar/broth method. It was found that 18 (94.7%) strains of Klebsiella pneumoniae, 21 (84.0%) strains of S aureus and 25 (65.7%) strains of coagulase-negative Staphylococci were positive for slime or slime-like substances by Congo red agar/broth method. A total of 41.0% of the strains positive by Christensen's method and 15.2% of the strains by dye elution technique were found to be more adherent organisms and that have the potential to form biofilms. Only the gram-positive organisms showed nonspecific agglutination with latex suspension. CONCLUSION : Among the various phenotypic methods compared in this study the Congo red agar/broth method is a simple, economical, sensitive, and specific method that can be used by clinical microbiology laboratories for screening the strains for the presence of slime or slime-like substances.

Iron Indices in Patients with Functional Anemia in Chronic Kidney Disease.

BACKGROUND: Despite high ferritin level, HDCKD patients may have functional iron deficiency even after intravenous iron (iv) therapy. The aim of this study was to test the hypothesis that lowered serum transferrin level may contribute to functional anemia and erythropoietin hypo responsiveness by the failure to transport accumulated tissue iron to the relevant target tissue. MATERIALS AND METHODS: The study subjects were divided into four groups. Group-A: HDCKD Patients receiving iv iron (n=290). Group-B: Patients not initiated on to hemodialysis (NDCKD), and received oral iron (n=38). Group-C: HDCKD patients with erythropoietin hypo responsiveness (n=9). Group-D: Healthy controls (n=36). The group-A, patients were sub-divided into five groups (A-1 to A-5) based on their serum ferritin levels. RESULTS: Serum ferritin and tissue iron levels in group-A and C patients were significantly greater than the group-D(p<0.0001) and group-B patients(p<0.001). Transferrin level of group-A and C showed lowered values and consequently a higher %TSAT when compared to group-D. The percent of patients with iron overload was 2.6%, 31%, and 44% in group-B, group-A and group-C respectively. Serum transferrin level significantly correlated with TIBC in group-A and B patients (p<0.0001;p<0.05 respectively). Transferrin level significantly correlated with TIBC in all subgroups of HDCKD(p<0.05) with the exception in subgroup-A2 and with hemoglobin in subgroups A3 (p<0.05) and A5(p<0.01) respectively. CONCLUSIONS: The lowered transferrin level prevents the proper transport of the iron to the hematopoietic sites, which may be a reason for the low hemoglobin synthesis and also for the development of erythropoietin hypo responsiveness in some of the dialysis patients.

Importance of Serum Copper and Vascular Endothelial Growth Factor (VEGF-A) Levels in Postmenopausal Bleeding.

The main aim of this study is association of serum copper and vascular endothelial growth factor (VEGF-A) in postmenopausal bleeding (PMB) patients. Blood samples were collected from female patients suffering with postmenopausal bleeding (n = 50) as well as healthy females as controls (n = 50). Serum copper levels were estimated by spectrophotometric method and serum VEGF-A by ELISA technique and compared with ultrasonographic measurement of endometrial thickness in both patients and controls. A significant increase in serum copper levels and an almost twofold increase in serum VEGF-A was observed in DUB patients when compared with controls. Correlation (r) between serum VEGF-A levels and endometrial thickness was 0.96. Odds ratio for copper, VEGF-A and combination of copper and VEGF-A was 0.0426, 0.0947 and 0.0313 respectively, in all these cases odds ratio was <1. The abnormal angiogenesis in PMB could be due to increased serum copper levels,which in turn stimulates factors like VEGF-A, thereby causing an increase in endometrial growth.

The effect of lithium on the adrenoceptor-mediated second messenger system in the rat brain.

OBJECTIVE: Lithium remains the most widely used treatment for bipolar disorder; however, the molecular mechanisms underlying its therapeutic actions have not been fully elucidated. We studied the in-vivo effect of lithium on the density of alpha-adrenoceptor (alpha-AR) and beta-AR subtypes and linked second messenger systems in the rat brain. METHODS: The densities of alpha(1)-ARs, alpha(2)-ARs, and beta(1)-ARs and beta(2)-ARs in the cortex and cerebellum of rats treated with lithium (0.4%), orally, for 30 days were measured using [(3)H]prazosin, [(3)H]clonidine and [(3)H]CGP-12177, respectively. The activity of adenylyl cyclase (AC) and levels of inositol trisphosphate (IP3), both second messengers linked to these receptors, were estimated using [(3)H]ATP and [(3)H]myoinositol, respectively. RESULTS: A significant decrease in the densities of cortical alpha(1)-ARs (85%, p < 0.0001), alpha(2)-ARs (50%, p < 0.0001), beta(1)-ARs (26%, p < 0.0001) and beta(2)-ARs (25%, p < 0.0001) was observed after lithium treatment. However, only the density of alpha(1)-ARs was significantly decreased (25%, p < 0.0001) in the cerebellum. The affinity of [(3)H]prazosin for cerebellar alpha(1)-ARs was increased. A small, but statistically significant, increase (19%, p < 0.0001) in the density of total beta-ARs was seen in the cerebellum, without altering the affinity of the radioligand for these receptors. Basal AC activity was not altered in the lithium-treated rat cortex. However, the norepinephrine-stimulated AC activity, which represents alpha(2)-AR-linked and beta-AR-linked AC, was significantly increased (66%, p < 0.0001). Both basal IP3 formation and norepinephrine-stimulated IP3, which represents alpha(1)-AR-linked phospholipase C activity, were significantly decreased (50%, p < 0.0001) in the lithium-treated rat cortex. CONCLUSION: Our results suggest that long-term administration of lithium treatment downregulates the cortical, but not cerebellar, alpha(1)-ARs, alpha(2)-ARs, beta(1)-ARs and beta(2)-ARs. Thus, it may be concluded that lithium induces region-specific and differential functional downregulation of alpha-AR and beta-AR subtypes in the rat brain.