Melioration in Anti-staphylococcal Activity of Conventional Antibiotic(s) by Organic Acids Present in the Cell Free Supernatant of Lactobacillus paraplantarum.

In view of emerging drug resistance in pathogens, there is a need to explore alternative strategies to combat infections. Use of probiotics is one such option. In this regard, efficacy of Lactobacillus plantarum has been reported against Staphylococcus aureus. Here, we propose that cell free supernatant (CFS) of Lactobacillus paraplantarum when used in combination with conventional antibiotics viz. ampicillin and oxacillin [to which the methicillin resistant Staphylococcus aureus (MRSA) strains were originally resistant] reduce the minimum inhibitory concentrations of these antibiotics, rendering the combination either synergistic or additive against the tested MRSA strain. The anti-staphylococcal activity was observed to be due to organic acids (acetic acid and lactic acid as confirmed by HPLC analysis) present in the CFS, as neutralization of the CFS with an alkali, sodium hydroxide (NaOH), caused the complete abrogation of its activity. The role of H2O2 and bacteriocin present in the CFS was also ruled out. The findings of this study suggest that cell free supernatant and ampicillin/oxacillin combination(s) might help in rejuvenating the use of conventional anti-staphylococcal antibiotics for the treatment of multi-drug resistant strains.

Evaluation of Candida Scoring Systems to Predict Early Candidemia: A Prospective and Observational Study at a Tertiary Care Hospital, Uttarakhand.

BACKGROUND: Candidemia in critically ill patients is usually a severe and life-threatening condition. Furthermore, due to its nonspecific presentation, it is difficult to diagnose leading to delayed treatment, prolonged hospitalization, and increased health-care costs with increase in morbidity and mortality. OBJECTIVES: In view of lack of data on "Candida scoring systems," this study was designed to evaluate the effectiveness of these scoring systems in predicting the development of candidemia among the Intensive Care Unit patients. MATERIALS AND METHODS: The "Candida score" was calculated at the onset of systemic inflammatory response syndrome, sepsis, or shock. Various scoring systems were compared using the area under the receiver operating characteristic curve. RESULTS: Among all three bedside risk scoring systems to predict candidemia both Leon score and Wenzel score offered significant discrimination between candidemic and noncandidemic patients with P = 0.000 and 0.001, respectively. The area under the curve for the scoring systems was 0.946 (95% confidence interval [CI] = 0.89-1) and 0.818 (95% CI = 0.687-0.949). CONCLUSION: Leon scoring system was found to have highest specificity, diagnostic accuracy, and positive likelihood ratio among all. Thus, we might conclude that a Leon score of ≥2.5 was most suitable for diagnosis of candidemia with significant accuracy and shortening of turnaround time when compared to the gold standard of blood culture. To the best of our knowledge, this is the first report on the subject.

Characterization of the Antimicrobial Peptide Penisin, a Class Ia Novel Lantibiotic from Paenibacillus sp. Strain A3.

Attempts to isolate novel antimicrobial peptides from microbial sources have been on the rise recently, despite their low efficacy in therapeutic applications. Here, we report identification and characterization of a new efficient antimicrobial peptide from a bacterial strain designated A3 that exhibited highest identity with Paenibacillus ehimensis. Upon purification and subsequent molecular characterization of the antimicrobial peptide, referred to as penisin, we found the peptide to be a bacteriocin-like peptide. Consistent with these results, RAST analysis of the entire genome sequence revealed the presence of a lantibiotic gene cluster containing genes necessary for synthesis and maturation of a lantibiotic. While circular dichroism and one-dimension nuclear magnetic resonance experiments confirmed a random coil structure of the peptide, similar to other known lantibiotics, additional biochemical evidence suggests posttranslational modifications of the core peptide yield six thioether cross-links. The deduced amino acid sequence of the putative biosynthetic gene penA showed approximately 74% similarity with elgicin A and 50% similarity with the lantibiotic paenicidin A. Penisin effectively killed methicillin-resistant Staphylococcus aureus (MRSA) and did not exhibit hemolysis activity. Unlike other lantibiotics, it effectively inhibited the growth of Gram-negative bacteria. Furthermore, 80 mg/kg of body weight of penisin significantly reduced bacterial burden in a mouse thigh infection model and protected BALB/c mice in a bacteremia model entailing infection with Staphylococcus aureus MTCC 96, suggesting that it could be a promising new antimicrobial peptide.

Scrub typhus in Uttarakhand & adjoining Uttar Pradesh: Seasonality, clinical presentations & predictors of mortality.

BACKGROUND & OBJECTIVES: Scrub typhus is a re-emerging mite-borne rickettsiosis, which continues to be underdiagnosed, with lethal consequences. The present study was conducted to determine the seasonality, clinical presentation and predictors of mortality in patients with scrub typhus at a tertiary care teaching hospital in northern India. METHODS: Scrub typhus was suspected in patients attending the hospital as per the standard case definition and serological evidence was obtained by performing an IgM ELISA. RESULTS: A total of 284 patients with scrub typhus from urban and rural areas were seen, predominantly from July to November. The most common clinical presentation was a bilateral community-acquired pneumonia (CAP), which resembled pneumonia due to atypical pathogens and often progressed to acute respiratory distress syndrome (ARDS). An acute undifferentiated febrile illness (AUFI) or a febrile illness associated with altered sensorium, aseptic meningitis, shock, abdominal pain, gastrointestinal bleeding or jaundice was also seen. Eschars were seen in 17 per cent of patients, and thrombocytopenia, transaminitis and azotaemia were frequent. There were 24 deaths (8.5%) caused predominantly by ARDS and multi-organ dysfunction. The mortality in patients with ARDS was high (37%). ARDS [odds ratio (OR)=38.29, 95% confidence interval (CI): 9.93, 147.71] and acute kidney injury (OR=8.30, 95% CI: 2.21, 31.21) were the major predictors of death. INTERPRETATION & CONCLUSIONS: The present findings indicate that scrub typhus may be considered a cause of CAP, ARDS, AUFI or a febrile illness with multisystem involvement, in Uttarakhand and Uttar Pradesh, especially from July to November. Empiric therapy of CAP may include doxycycline or azithromycin to ensure coverage of underlying unsuspected scrub typhus.

Needle Stick Injury Among Healthcare Workers in a Tertiary Care Setting in Dehradun, Sub-Himalayan Region: A Four-Year Record-Based Study.

INTRODUCTION: Needlestick injuries (NSIs) represent a significant occupational health risk in healthcare settings. These injuries, caused by contaminated sharps such as needles, vials, and scalpel blades, can lead to percutaneous exposure to infectious materials. Despite the severity of NSIs, they often go unreported, highlighting a critical gap in occupational safety protocols. AIMS: This study aimed to investigate the occurrence of NSIs among healthcare workers (HCWs) by sex, profession, and working areas. It also sought to explore the underlying reasons for these injuries and the factors contributing to their underreporting. METHODOLOGY: Adhering to the RECORD guidelines (Reporting of studies Conducted using Observational Routinely Collected Data), this record-based study involved a retrospective analysis of reported NSIs. Data were collected from voluntary reports by HCWs who experienced NSIs or exposure to potentially infectious materials such as blood and body fluids. Statistical analysis was conducted using IBM SPSS Statistics for Windows, Version 16 (Released 2007; IBM Corp., Armonk, New York) and Microsoft Excel 2010 (Microsoft Corporation, Redmond, Washington). RESULTS: Data from 142 participants indicated a higher proportion of females experiencing NSIs compared to males, with rates of 57.7% pre-COVID and 60.6% during COVID. There were notable shifts in NSI rates across professions, with increases observed among staff nurses and ward attendants/helpers. Analysis of injury circumstances revealed a decrease in sampling procedure-related injuries but an increase during intravenous procedures and biomedical waste segregation. Injuries occurring on the right-hand index finger decreased from 52.1% pre-COVID to 31% during COVID, while those on the left-hand index finger increased from 19.7% pre-COVID to 39.4% during COVID. Statistically significant associations were found between the injury site and the place of occurrence (p=0.021). Healthcare professionals commonly cleansed the site with disinfectants and used personal protective equipment (PPE) kits, with increased PPE usage noted during the COVID-19 pandemic. These findings emphasize the evolving dynamics of NSIs among HCWs and underscore the importance of tailored preventive measures during pandemics.

Comparative performance of biofire pneumonia panel and standard culture-based methods for diagnosing pneumonia in critically ill patients: Impact on antibiotic stewardship.

INTRODUCTION: Lower respiratory tract infections (LRTIs) are a common cause of morbidity and mortality worldwide. Accurate identification of the pathogens causing LRTIs is crucial for ensuring of diagnostic and antibiotic stewardship. The Biofire Pneumonia Panel (BFPP) is a molecular diagnostic test that allows rapid detection of various bacterial and viral pathogens. In this study, we compared the performance of BFPP with standard culture methods for the detection of pathogens. MATERIALS AND METHODS: Respiratory samples from 70 patient with suspected LRTIs were tested using both BFPP and standard culture methods. The distribution of isolated bacterial pathogens was analyzed, and the sensitivity and specificity of BFPP were calculated. Additionally, the performance of BFPP in detecting antimicrobial resistance genes was evaluated. The results were compared with those obtained from VITEK-2 antimicrobial susceptibility testing and culture-based methods. RESULTS: Among the suspected LRTI cases, BFPP identified a single pathogen in 32.8% of cases and multiple pathogens in 40% of cases. The standard culture method detected a single pathogen in 47.1% of cases. BFPP showed a sensitivity of 93.9% and a specificity of 45.9% for the total sample. The performance of BFPP in detecting antimicrobial resistance genes varied for different pathogens with overall sensitivity of 40.1% and specificity of 95.9%. CONCLUSION: The Biofire Pneumonia Panel (BFPP) demonstrated high sensitivity for several bacterial pathogens, indicating its potential as a rapid diagnostic tool. However, its performance varied for different microorganisms, and it had limitations in detecting certain pathogens and antimicrobial resistance genes for which still required more further studies to explore different resistance gene mechanism that can be incorporated in this panel in future. The BFPP can complement standard culture methods as a rapid tool in the diagnosis of LRTIs.

Pro- and anti-inflammatory cytokines signatures at different severity of dengue infection.

Context: Dengue disease severity and progression are determined by the host immune response, with both pro- and anti-inflammatory cytokines are key mediators. Aims: To study pro- and anti-inflammatory cytokines across dengue severity and as a biomarker for predicting severe dengue infection. Settings and Design: Hospital-based cross-sectional study was conducted on 125 dengue-positive subjects across the 5-60 years age group of either gender in 2022. Methods and Materials: Haematological parameters and blood samples were drawn to measure cytokines IL6, IL-10 and TNF alpha using the ELISA technique. Statistical Analysis: One-way ANOVA and the Kruskal - Wallis test were used to compare the dependent variables across categories of the dengue spectrum. Receiver operating characteristic curve was drawn to calculate the predictability of the cytokines as a predictor of severe dengue. A P < 0.05 was considered significant. Results: 34.4% of cases had severe dengue infection with 53.2% of severe cases reported in >40 years of age. Only IL-6 levels significantly increased (P < 0.01) across the spectrum of dengue infection across age groups >20 years with a consistent and significant fall in platelet levels (P < 0.01). The accuracy of IL-6 to predict severe dengue was 74.4% and platelet count was 16.2%. Conclusions: Only IL-6 cytokine levels were significantly increased across the spectrum of dengue infection observed in age >20 years and can significantly predict the probability of severe dengue by 74% (sensitivity 81.4%). A significant decrease in platelet values is consistent with the severity but is not a good predictor for severe dengue infection.

An Observational Study of Fungal Infections in COVID-19: Highlighting the Role of Mucormycosis in Tertiary Healthcare Settings.

Background Fungal infections, especially mucormycosis, have remarkably surged during the coronavirus disease 2019 (COVID-19) era, especially during the second wave peak of the pandemic raising the concern of the clinicians for the admitted patients. Steroid therapy, diabetes, and other immunocompromised states are more commonly associated with COVID-19-associated mucormycosis (CAM). Aim and objective The aim of this study is to ascertain the prevalence of fungal infections amidst the second wave of the COVID-19 pandemic and discern the associated risk factors. Materials and methods During the second peak of COVID-19, samples were received in the microbiology laboratory from all clinically suspected mucormycosis patients. These samples underwent processing for potassium hydroxide (KOH) wet mount, fungal culture on Sabouraud's dextrose agar (SDA) medium, and COVID-19 reverse transcription-polymerase chain reaction (RT-PCR) testing. All relevant clinical and associated risk factors were tabulated and analyzed. Results Among the 107 suspected cases of mucormycosis, 39 (36.4%) were confirmed positive for COVID-19 via RT-PCR, while 68 (63.6%) tested negative. Males exhibited a predominant infection rate, with the rhinocerebral system being the most commonly affected site. Significantly higher mortality rates were observed in COVID-19-associated mucormycosis (CAM) patients (33.4%) compared to those without COVID-19 (5.9%), with a notable p-value of 0.0005. CAM patients also demonstrated a higher frequency of ICU admissions (77%) compared to non-COVID-19-associated mucormycosis patients (21.4%), a statistically significant finding (p-value of 0.007). Additionally, immunocompromised states, diabetes, and the administration of oxygen therapy were identified as significant risk factors in CAM (p < 0.05). Notably, mucormycosis accounted for the majority of fungal isolates (48.27%) among COVID-19 patients. Conclusion Mucormycosis infection is more commonly seen in COVID-19-infected patients as compared to non-COVID-19 patients, especially with comorbidities such as diabetes mellitus, steroid usage, and other immunocompromised states.

Phenotypic characterization and antifungal susceptibility profile of Candida isolates from women with vulvovaginal candidiasis at a community health center linked to a teaching institution in the Sub-Himalayan region of North India.

Introduction: Vulvovaginal candidiasis (VVC) is considered a common gynecological problem among females of reproductive age group. 70-75% of women report having had candidal vulvovaginitis at some point in their lifetimes and 40-50% suffer recurrent candidal vulvovaginitis. Objectives: This study aims to identify the Candida species involved in VVC and to determine their antifungal susceptibility pattern. Materials and Methods: The present study was a cross-sectional study conducted on 257 females (18-55 yr) with complaints of abnormal vaginal discharge. For detection of Candida, the swab samples were subjected to Gram stain, 10% KOH mount, and culture on Sabouraud dextrose agar (SDA). Candida species identification was done by subculturing Candida isolates onto CHROMagar, corn meal agar (Himedia), and further confirmation was done by MALDI-TOF MS. Antifungal testing was done using the disk diffusion method. Results: A total of 257 females with complaints of abnormal discharge were enrolled in this study. Out of 257, C. albicans 37 (58.7%) and 26 (41.3%) isolates were identified as non-albicans Candida. Out of 63 positive cases, a maximum number of study subject belongs to the age group 26-35 years (50.8%). Along with vaginal discharge, itching (65.37%) is the most common complaint. VVC was found to be most predominant in patients with prolonged antibiotic therapy (38.1%), and in pregnant females (15.9%). Conclusion: Understanding the emerging fungal pathogens and their drug susceptibility patterns is essential for the effective management of infections. Drug resistance can lead to treatment failure and highlights the need for alternative treatment options or strategies.

Pharmacokinetic assessment of cefpodoxime proxetil in diabetic rats.

Purpose: In diabetes, multi-organ level dysfunction arising from metabolic complications is reported to influence the pharmacokinetics (PK) profile of many drugs. Hence, the present study was planned in rats to evaluate the effect of diabetes on the PK profile of cefpodoxime, a widely prescribed oral antibiotic. Method: PK profile of cefpodoxime was assessed after oral administration of cefpodoxime proxetil (10 and 20 mg/kg) and intravenous (i.v) administration of cefpodoxime sodium (10 mg/kg) in normal and streptozotocin induced diabetic rats. To evaluate the impact of diabetes on oral absorption and serum protein binding, in situ intestinal permeability and in vitro serum protein binding studies were performed for cefpodoxime using Single Pass Intestinal Perfusion model (SPIP) and ultracentrifugation technique, respectively. Result: In diabetic rats, there was significant (p < 0.01) decrease in maximum concentration (Cmax) and area under the curve (AUC) of cefpodoxime by both oral and intravenous route, which was attributed to augmented clearance of cefpodoxime. There was no change in the time to achieve Cmax (Tmax) suggesting no alteration in oral absorption which was further confirmed through unaltered intestinal permeability in diabetic rats. The protein binding in diabetic rats also remained unchanged, indicating no influence of protein binding on elevated clearance. Conclusion: The plasma exposure of cefpodoxime, a renally eliminated drug was significantly lowered in diabetic rats due to enhanced glomerular filtration. However, this observation needs to be confirmed through well controlled clinical trials.

Insights on Quercetin Therapeutic Potential for Neurodegenerative Diseases and its Nano-technological Perspectives.

The neurodegeneration process begins in conjunction with the aging of the neurons. It manifests in different parts of the brain as Aß plaques, neurofibrillary tangles, Lewy bodies, Pick bodies, and other structures, which leads to progressive loss or death of neurons. Quercetin (QC) is a flavonoid compound found in fruits, tea, and other edible plants have antioxidant effects that have been studied from subcellular compartments to tissue levels in the brain. Also, quercetin has been reported to possess a neuroprotective role by decreasing oxidative stress-induced neuronal cell damage.The use of QC for neurodegenerative therapy, the existence of the blood-brain barrier (BBB) remains a significant barrier to improving the clinical effectiveness of the drug, so finding an innovative solution to develop simultaneous BBB-crossing ability of drugs for treating neurodegenerative disorders and improving neurological outcomes is crucial. The nanoparticle formulation of QC is considered beneficial and useful for its delivery through this route for the treatment of neurodegenerative diseases seems necessary. Increased QC accumulation in the brain tissue and more significant improvements in tissue and cellular levels are among the benefits of QC-involved nanostructures.

Validity of International Classification of Diseases (ICD)-10 Diagnosis Codes for Identification of Acute Heart Failure Hospitalization and Heart Failure with Reduced Versus Preserved Ejection Fraction in a National Medicare Sample.

BACKGROUND: Heart failure (HF) is a leading cause of hospitalization in older adults. Medicare data have been used to assess HF outcomes. However, the validity of ICD-10 diagnosis codes (used since 2015) to identify acute HF hospitalization or distinguish reduced (heart failure with reduced ejection fraction) versus preserved ejection fraction (HFpEF) is unknown in Medicare data. METHODS: Using Medicare data (2015-2017), we randomly sampled 200 HF hospitalizations with ICD-10 diagnosis codes for HF in the first/second claim position in a 1:1:2 ratio for systolic HF (I50.2), diastolic HF (I50.3), and other HF (I50.X). The primary gold standards included recorded HF diagnosis by a treating physician for HF hospitalization, ejection fraction (EF)≤50 for heart failure with reduced ejection fraction, and EF>50 for HFpEF. If the quantitative EF was not present, then qualitative descriptions of EF were used for heart failure with reduced ejection fraction/HFpEF gold standards. Multiple secondary gold standards were also tested. Gold standard data were extracted from medical records using standardized forms and adjudicated by cardiology fellows/staff. We calculated positive predictive values with 95% CIs. RESULTS: The 200-chart validation sample included 50 systolic, 50 diastolic, 47 combined dysfunction, and 53 unspecified HF patients. The positive predictive values of acute HF hospitalization was 98% [95% CI, 95-100] for first-position ICD-10 HF diagnosis and 66% [95% CI, 58-74] for first/second-position diagnosis. Quantitative EF was available for ≥80% of patients with systolic, diastolic, or combined dysfunction ICD-10 codes. The positive predictive value of systolic HF codes was 90% [95% CI, 82-98] for EFs≤50% and 72% [95% CI, 60-85] for EFs≤40%. The positive predictive value was 92% [95% CI, 85-100] for HFpEF for EFs>50%. The ICD-10 codes for combined or unspecified HF poorly predicted heart failure with reduced ejection fraction or HFpEF. CONCLUSIONS: ICD-10 principal diagnosis identified acute HF hospitalization with a high positive predictive value. Systolic and diastolic ICD-10 diagnoses reliably identified heart failure with reduced ejection fraction and HFpEF when EF 50% was used as the cutoff.

Vitex negundo protects against cerebral ischemia-reperfusion injury in mouse via attenuating behavioral deficits and oxidative damage.

Global cerebral ischemia/reperfusion (I-R) injury often results in an irreparable brain damage like behavioral impairment and neuronal death. This neurological complication involves diverse intricate pathological mechanisms like oxidative stress, inflammation, and apoptosis. Recently, the therapeutic value of plant-based polyphenols has gained researcher's attention. The present study focused on the putative neuroprotective role of negundoside on behavioral and oxidative stress status in an experimental model of global cerebral ischemia and reperfusion-induced brain injury. Negundoside was isolated from the leaves of Vitex negundo Linn. by chromatography for investigating its possible neurobehavioral and neuropharmacological implications. Healthy Balb/C mice of either sex were subjected to 10 min of global cerebral ischemia (GCI) followed by 24-h reperfusion. Mice were pre-treated intraperitoneally with negundoside at varying doses (1, 3, 5, 10, and 15 mg/kg) 60 min before the induction of GCI. Mice were later subjected to a battery of behavioral tests for evaluating memory-related and motor abilities. Elevated plus maze (EPM) was used to determine the anxiety levels and short-term memory whereas motor abilities were evaluated by inclined beam-walking test, rotarod, and lateral push test. TBARS and reduced glutathione (GSH) content in brains were analyzed spectrophotometrically as oxidative stress markers. Behavioral study revealed enhanced anxiety-related responses and motor deficits in I-R injured mice. Additionally, GSH and TBARS levels were found to be altered following I-R-induced neuronal injury. Contrastingly, negundoside administration was able to alleviate the behavioral and biochemical alterations to the normal levels. Together, our findings provide preliminary evidence of neuroprotective role of negundoside against global cerebral ischemia and reperfusion-induced behavioral dysfunction and oxidative damage in mice brain.

Locally harvested Covid-19 convalescent plasma could probably help combat the geographically determined SARS-CoV-2 viral variants.

The only effective way to provide individuals with herd immunity against the novel coronavirus [SARS-CoV-2] is to administer an effective vaccine that will help check the current pandemic status. In India, the central drugs standard control organization (CDSCO) has granted the emergency-use authorization [EUA] to three vaccines namely, Covishield (live vaccine, Oxford AstraZeneca, United Kingdom being manufactured by the Serum Institute of India), Covaxin (inactivated vaccine, Bharat Biotech, India) and Sputnik V (live vaccine, Gamaleya, Russia). However, there is a rising need for the efficacy of the vaccines to be proven against the "SARS-CoV-2 viral variants." Also, human plasma is polyclonal in nature with an inherent propensity to identify multiple epitopes of either an antigen or pathogen. With this context in mind, the researchers hypothesize that using COVID-19 convalescent plasma [CCP] harvested from the locally recovered individuals [i.e. potential CCP donors] may be particularly beneficial in combating not only the founder SARS-CoV-2 virus but also the geographically determined SARS-CoV-2 variants among the regionally affected COVID-19 patients.

Factors affecting the serological testing of cadaveric donor cornea.

PURPOSE: The purpose of this study was to evaluate the serological profile of the eye donors and to study the influence of various factors on serological test results. METHODS: A cross-sectional, observational study was conducted, and data of 509 donors were reviewed from the records of eye bank from December 2012 to June 2017. Various details of donors analyzed included the age, sex of the donor, cause of death, source of tissue, time since blood collection after death, macroscopic appearance of blood sample, and details of discarded tissues. Serological examination of blood was performed for human immunodeficiency virus (HIV), hepatitis B virus, hepatitis C virus (HCV), venereal disease research laboratory (VDRL), and serology reports reactive or nonreactive were analyzed. RESULTS: Among the 509 donors, 295 (58%) were male, and 420 (82.50%) belonged to age group ≥60 years. Most donors (354, 69.5%) died due to cardiac arrest. Macroscopically, sera were normal in the majority of 488 (95.9%) cases. Among 509 donors, 475 (93.3%) were nonreactive, 12 (2.4%) donors were found to be reactive to hepatitis B surface antigen (HBsAg), and 1 (0.2%) was reactive to HCV, but no donor serology was reactive to HIV or VDRL. Twenty-one (4.12%) donors' sera were not fit for serological testing. Among all donors, 475 (93.32%) donors were accepted and 34 (6.67%) were rejected or discarded on the basis of serological testing. Cause of death and macroscopic aspect of sera influenced the serological results in a highly significant manner (P = 0.00). Acceptance or rejection of the donor was significantly influenced by the serological results of the donor (P = 0.00). CONCLUSION: The seroprevalence among eye donor for HBsAg and HCV was 12 (2.4%) and 1 (0.2%), respectively. Factors such as cause of death and macroscopic aspect of sera influence the serological results. Time since blood collection or sampling will not show any impact on viral serological results if postmortem sampling will be done in < 10 hours(h) after death which can improve the safety and utility of the donor cornea.

Evaluation of haematological and volume, conductivity and scatter parameters of leucocytes for aetiological diagnosis of undifferentiated fevers.

Background: In this follow-up study, we aimed to establish the cut-off values of the volume, conductivity and scatter (VCS) parameters of leucocytes that significantly differ in dengue fever, malaria, scrub typhus and enteric fever, as described in our pilot study. Methods: A prospective observational case-control study was undertaken on patients with an established diagnosis of acute malaria (n=476), dengue fever (n=927), scrub typhus (n=425), bacterial sepsis (n=1598) or enteric fever (n=885) and the haematological and VCS data obtained by the Coulter LH 750 analyser were compared with controls. Results: Malaria significantly altered the platelet count and plateletcrit. The cut-offs for the monocyte volume, lymphocyte volume, neutrophil volume, neutrophil volume distribution width, lymphocyte volume distribution width, monocyte volume distribution width and neutrophil conductivity distribution width had 90% negative predictive values (NPVs) for malaria. Except for the proportion of monocytes, lymphocyte scatter distribution width and platelet distribution width, a >70% positive (PPV) and negative predictive value (NPV) for dengue was noted in all the haematological and VCS parameters analysed. While the cut-offs determined in scrub typhus had a good (>80%) PPV and NPV, a significant difference was seen in neutrophil, monocyte and lymphocyte conductivities, but with low sensitivity, specificity, PPV and NPV in enteric fever. Conclusions: Aetiology-specific changes occur both in numbers as well as in VCS properties in acute fevers. Formulae and/or algorithms need to be utilized to realize the full potential of the VCS technology.

A Hospital-Based Retrospective Comparative Study of Complications, Outcomes, Clinical and Laboratory Parameters of Malaria with and without Neurological Involvement.

BACKGROUND & OBJECTIVES: Classically associated with Plasmodium (P.) falciparum, neurological complications in severe malaria is associated with increased morbidity and mortality. However, reports implicate the long considered benign P. vivax for causing severe malaria as well. We aimed to analyse the cerebral complications in malaria, and study if there is a species-related difference in the presentation and outcomes. METHODS: We retrospectively compared patients with malaria hospitalised from 2009-15, with (n=105) and without (n=1155) neurological involvement regarding outcomes, complications, demographic attributes, clinical features, and laboratory parameters. Subsequently, the same parameters were studied in those with cerebral malaria due to mono-infections of P. vivax or P. falciparum and their co-infection. RESULTS: Cerebral malaria was observed in 8.3% (58/696), 7.4% (38/513) and 17.6% (6/51) of P. vivax, P. falciparum and combined plasmodial infections respectively. Those with cerebral malaria had significantly (p<0.05) longer hospitalisation, delayed defervescence, required mechanical ventilatory support and dialysis despite comparable levels of azotemia and renal insufficiency, and adverse outcomes compared to non-cerebral malaria. Severe thrombocytopenia, respiratory distress and mechanical ventilation were significantly (p<0.05) associated with P. vivax cerebral malaria. CONCLUSIONS: The plasmodial species are comparable in clinical and laboratory parameters and outcomes in cerebral malaria in isolation and combination (p>0.05). P. vivax is emerging as the predominant cause of cerebral malaria, and its virulence is comparable to P. falciparum.

Quantitative and volume, conductivity and scatter changes in leucocytes of patients with acute undifferentiated febrile illness: a pilot study.

BACKGROUND: A single diagnostic test for acute undifferentiated febrile illnesses (AUFI) is elusive. This pilot study was undertaken on the premise that leucocytes, being the main cells of defence, undergo quantitative, structural and functional changes in AUFI. We evaluated the potential of volume, conductivity and scatter (VCS) parameters of leucocytes, generated with the haemogram report by the Coulter auto-analyzer, in differentiating the common etiologies of AUFI. METHODS: The haematological and VCS data obtained from 800 controls and 200 cases of AUFI (50 cases each of acute malaria, dengue, scrub typhus and enteric fever) were retrieved for analysis. RESULTS: The cases and controls differed significantly with respect to relative numbers and the VCS parameters of neutrophils, lymphocytes and monocytes (p<0.05). The neutrophil and lymphocyte were significantly voluminous in acute malaria and scrub typhus as compared to dengue and enteric fevers (p<0.05). Enteric fever significantly enhanced the conductivity of neutrophils as compared to other subgroups while lymphocyte conductivity significantly differed from dengue and scrub typhus. Lymphocyte and neutrophil scatter values in malaria and scrub typhus were comparable but differed significantly from that in enteric fever. CONCLUSIONS: Etiology-specific changes occur in leucocytes, both in numbers and their VCS properties which can be identified without additional cost.

The Effects of Cryomilling CNTs on the Thermal and Electrical Properties of CNT/PMMA Composites.

In this study, the cryomilling of carbon nanotubes (CNTs) was carried out to accomplish better dispersion without using any hazardous chemicals. Accordingly, different samples of CNTs were prepared by varying the milling speed (10, 20, and 25 Hz) and time (5, 10, and 15 min) and incorporated into the poly(methyl methacrylate) (PMMA) matrix. The changes of the morphology were analyzed by utilizing a field emission scanning electron microscope (FESEM) and a high-resolution transmission electron microscope (TEM). Qualitative analysis of the cryomilled CNTs was carried out using Raman spectroscopy, and their surface area was determined via Brunauer⁻Emmett⁻Teller (BET) analysis. Subsequently, thermogravimetric analysis was conducted to evaluate the thermal properties, whereas the surface resistivity and electromagnetic interference shielding effectiveness for the electrical conductivity were also examined. It was observed that the composite with Cr-20-10 showed better thermal stability and lower resistivity in comparison to the others because, as the cryomilling time and frequency increased the distribution, dispersion and surface area also increased. Consequently, a better interaction between CNTs and PMMA took place.

Topical Delivery of Protein and Peptide Using Novel Cell Penetrating Peptide IMT-P8.

Skin, being the largest organ of the body, is an important site for drug administration. However, most of the drugs have poor permeability and thus drug delivery through the skin is very challenging. In this study, we examined the transdermal delivery capability of IMT-P8, a novel cell-penetrating peptide. We generated IMT-P8-GFP and IMT-P8-KLA fusion constructs and evaluated their internalization into mouse skin after topical application. Our results demonstrate that IMT-P8 is capable of transporting green fluorescent protein (GFP) and proapoptotic peptide, KLA into the skin and also in different cell lines. Interestingly, uptake of IMT-P8-GFP was considerably higher than TAT-GFP in HeLa cells. After internalization, IMT-P8-KLA got localized to the mitochondria and caused significant cell death in HeLa cells signifying an intact biological activity. Further in vivo skin penetration experiments revealed that after topical application, IMT-P8 penetrated the stratum corneum, entered into the viable epidermis and accumulated inside the hair follicles. In addition, both IMT-P8-KLA and IMT-P8-GFP internalized into the hair follicles and dermal tissue of the skin following topical application. These results suggested that IMT-P8 could be a potential candidate to be used as a topical delivery vehicle for various cosmetic and skin disease applications.