A Study on Virulence Determinants of Clinical Isolates of Candida Species and Their Correlation with Antifungal Resistance in a Tertiary Care Hospital in Eastern India

Candida Species are normal flora of skin and mucosa of gastrointestinal tract (GIT) and genital region. Usually they invade host tissue and cause diseases in patients with impaired immunity. OBJECTIVE: The aim of this study is to determine virulence factors expressed by various clinical isolates of different Candida species and to correlate with their antifungal resistance pattern. Methods: A total of 120 Candida isolates obtained from different clinical samples and speciated by standard microbiological laboratory protocol. Haemolysin, phospholipase and esterase activities were detected by growth characteristics on SDA with blood, egg yolk agar medium and Tween- 80 opacity test medium respectively. Adherence was seen microscopically over buccal epithelial cell and biofilm formation was detected visually with 2% Safranin staining. Antifungal susceptibility was detected by disk diffusion method using Mueller Hinton Agar supplemented with glucose and methylene blue (As per CLSI guideline). Results: Out of 120 isolates, 51(42.50%) were C.albicans and 69 (57.5%) were Non albicans Candida (NAC) including C.tropicalis 32(26.67%), C. guilliermondii 21(17.50%), C. parapsilosis 12(10%) and C. glabrata 04(3.33%). Haemolysin, phospholipase and esterase activities were detected in 110 (91.67%), 95 (79.17%) and 82 (68.33%) respectively. Adherence seen in 98 (81.67%) and in 59 (49.17%) cases biofilm had formed. The antifungal susceptibility patterns revealed that 81 (67.5%) were susceptible to Fluconazole and 72 (60%) were susceptible to Itraconazole, whereas 100% were susceptible to Voriconazole and Amphotericin B. Conclusion: Detection of virulence factor will help in better understanding of the changing behavioural patterns of Candida isolates and open doorways to better management and assessing prognosis of the patients.

Profile of Disease in Leprosy Patients Diagnosed in a Tertiary Care Centre in Kolkata during 2014-2017

Study of profile of leprosy in a Tertiary care Centre in an Urban Metropolitan settings has important epidemiologically relevant messages. This study has been carried out to find out the profile of leprosy patients attending a Tertiary Care Centre at Kolkata and to interpret data in respect to different epidemiological variables. The cases attending R.G. Kar Medical College for the year 2014-2017 and who were referred to Microbiology Department for slit skin smear examination were studied. Their profile analyzed according to age, sex, type of disease, past treatment, presentation of disease, deformity status, presence of reactions, bacteriological status of the patients and treatment history. Diagnosis was based on the clinical assessment and slit skin smear for Bacteriological Index (BI) as well as Morphological index (MI). Among 2380 cases 720 (30.25%) were females and 1660 (69.75%) were males. Maximum patients (74%) belong to upper lower (Class IV) socio-economic class. 20-40 years age group was reported as peak age of onset with average onset of age as 35 years, 7.56% were children below 15 years of age. Maximum patients (63.87%) were in reproductive age group. Multibacillary (BL/LL) cases are more prevalent (71.47%). 21% of cases were bacteriologically positive with 10.9% having >3+ BI. MI upto 25% was observed in 92% of 360 smears examined for this purpose. Proportion of type 1 reaction was more than type 2 in both males and females. 440 (18.5%) had reactions, 12.6% had type 1 and 5.9% type 2 reactions. Reactions in females were more associated with pregnancy/ lactation. 24% of cases had grade I disability whereas grade II disability was present in 14% of cases which indicates delayed diagnosis and inadequate management or both. Both grade 1 and grade 2 were more in males. Leprosy was found to be more prevalent in males, more prevalent in upper lower (Class IV) socioeconomic class and more commonly of borderline type. Strategy need to be fine tuned to improve early detection, more focus on females during the reproductive years and appropriate intervention to reduce overall morbidity and deformity/disability

Enteric fever in an HIV/AIDS patient: atypical manifestations

Bloodstream infections with Salmonella typhi, is uncommon in human immunodeficiency virus [HIV]-infected persons. The symptoms in such patients are often non-specific and have a rather insidious onset and progression. We report a patient with sepsis and lower limb gangrene due to Salmonella typhi infection in an HIV-infected patient.

Diagnostic evaluation of extrapulmonary tuberculosis by fine needle aspiraton (FNA) supplemented with AFB smear and culture.

One hundred forty-three patients, 72 males and 71 females, with extrapulmonary tuberculosis were aspirated and subjected to cytological (Ziehl-Neelsen stain) examination and culture in Lowenstein-Jensen media. Routine haematological examination and Mantoux test were done in all the cases, x-ray chest in 112, skeletal x-ray in 3 relevant cases and sputum was examined for AFB in 16 cases where pulmonary tuberculosis was associated/suspected with extrapulmonary tuberculosis. HIV status was evaluated in 51 cases and 9 (7.64%) were seropositive. FNA cytology in 102 cases (71.3%) had caseating epithelioid granulomas while smear for AFB was positive in 57 cases (39.8%). Both culture and smear were positive in 29 (20.2%) cases. Combining both smear and culture yielded positive results in 47.5% cases. It was observed that AFB positivity was higher in untreated patients and with HIV positive cases. Further more, the triad of FNAC, AFB smear and culture were cheaper, foolproof and confirmatory than costlier tests like TB IgG, IgM, RTPCR and BACTEC.

Does pentoxifylline find a place in the armementarium of leprologists in type II reaction?

Alternative therapeutic interventions in Type II lepra reaction are being considered following serious problems associated with the use of steroids and thalidomide. Pentoxifylline (PTX) has been used in Type II reaction with varying degrees of success. The results of a study on the use of this drug in a dose of 1200 mg per day for a period of 2 months in patients with ENL reaction are discussed. Five patients, one of whom was HIV positive--all with severe Type II reaction, were regularly evaluated for regression of inflammatory symptoms and clinical involution of ENL lesions while on PTX therapy and thereafter. It was found that PTX led to a total elimination of systemic symptoms within a week. ENL lesions regressed in two weeks. However, in one patient, lesions recurred after one month of therapy. It appears that PTX is well tolerated and could be used as an additional drug in the armamentarium of leprologists in the management of Type II reaction, especially in HIV co-infection, where long-term steroids are contraindicated. However, further studies to compare the effects of PTX with currently, widely used drugs for the treatment of ENL reaction are necessary.

Acquired (digital) fibrokeratoma.

Acquired digital fibrokeratoma is a rarely reported disorder, especially in the Indian literature. We report our findings on one such case, occurring at an unusual site.

Ceftriaxone therapy in ciprofloxacin treatment failure typhoid fever in children.

BACKGROUND & OBJECTIVES: The rapid spread of multidrug resistant (MDR) typhoid fever has posed a great challenge for the treatment of these cases the world over. After the emergence of chloramphenicol resistant Salmonella typhi strains, ciprofloxacin has become the drug of choice for the treatment of typhoid fever even in the paediatric age group. This study evaluated the role of ceftriaxone therapy in bacteriologically confirmed MDR typhoid cases who did not respond to 12-14 days of ciprofloxacin therapy. Attempts have also been made to investigate the in vitro susceptibility of isolated S. typhi strains to chloramphenicol, ciprofloxacin and ceftriaxone. METHODS: A total of 140 children, aged 3-10 yr, clinically diagnosed as having typhoid fever, without any clinical response after 12-14 days of ciprofloxacin therapy were screened for S. typhi by blood culture. In the bacteriologically positive children the treatment was changed to intravenous ceftriaxone for 14 days. The isolated strains of S. typhi were tested for in vitro antimicrobial susceptibility. RESULTS: Clinical and bacteriological cure was observed with intravenous ceftriaxone therapy in all the 32 bacteriologically positive patients. All isolated S. typhi strains were uniformly (100%) susceptible to ciprofloxacin and ceftriaxone but 50 per cent of the strains were resistant to chloramphenicol. The MIC values of chloramphenicol, ciprofloxacin and ceftriaxone ranged between 125-500, 0.0625-0.5 and < 0.0625 microgram/ml respectively. INTERPRETATION & CONCLUSION: The study indicates that although the S. typhi strains were susceptible to ciprofloxacin in vitro, the patients did not respond clinically and bacteriologically to ciprofloxacin therapy. Hence, ciprofloxacin may not represent a reliable and useful option for treating MDR typhoid fever; ceftriaxone may be an effective alternative for the treatment of such cases.

Multi-drug resistant typhoid fever with diarrhea.

OBJECTIVE: To provide information about the characteristics of diarrheal stool in multi-drug resistant typhoid fever and observe the clinical course after treatment with furazolidone or ciprofloxacin. SETTING: Hospital based. SUBJECTS AND METHODS: Twenty one male children who were positive for multi-drug resistant S. typhi by blood and stool cultures, having diarrhea at the time of hospitalization comprised the subjects. Serum and stool electrolytes were estimated. Stool samples were also processed to detect established enteropathogens, leukocytes and red blood cells. Children were treated either with furazolidone or ciprofloxacin and evaluated till recovery. RESULTS: Mean (+/- SD) pre-admission duration of fever and diarrhea of these cases were 19.1 (+/- 5.6) and 15.8 (+/- 4.6) days, respectively. Stool character in 81% of the patients was watery with mean (+/- SD) volume of stool 51.4 (+/- 25.1) ml per kg body weight in the first 24 hours of observation. Leukocyte count varied between 20-49 per high power field in 66.7% stool samples. Occult blood was present in only 19% cases. Fecal red blood cells in high power field were detected in 52.4% cases. Mean fecal electrolytes (mmol/liter) were as follows: sodium-53.8, potassium-51.4, chloride-41.6 and total CO2-24.3. Most of the children (71.4%) had no dehydration and had normal serum electrolytes. The isolated strains of S. typhi were multi-drug resistant. These children were treated successfully either with furazolidone or ciprofloxacin. CONCLUSION: The stools of multi-drug resistant typhoid fever patients were watery with little blood. Their electrolyte contents were more similar to the diarrheal stool seen in shigellosis rather than cholera. Uncontrolled observations revealed that children recovered with furazolidone or ciprofloxacin therapy.

Microbial production of L-lysine: a review.

Microbial production of L-lysine has been reviewed with 251 references. The review includes different lysine producing microorganisms, their optimum cultural conditions, yield, assay and process of product recovery. It includes a discussion on the pathway of aspartate family of amino acid's biosynthesis in bacteria and its regulation. Achievements in this regard made through genetic engineering have also been included.

Recent kala-azar cases in India: isozyme profiles of Leishmania parasites.

In this study, three WHO recommended reference strains of Leishmania donovani (DD8), L. tropica (K27) and L. major (5 ASKH) were compared by isozyme analysis of 9 soluble enzymes using cellulose acetate electrophoresis (CAE). The isozyme profile of the three species showed distinct electrophoretic mobilities of the enzymes studied. This technique was applied to study the enzyme patterns of 8 clinical isolates of visceral leishmaniasis (VL) and two post kala-azar dermal leishmaniasis (PKDL) collected from the present epidemic (1990s to date). They were seen to be similar to the two isolates of the 1970s epidemic. Interspecies differences were observed in different reference Leishmania species whereas no intraspecies difference was observed amongst the clinical isolates (8VL + 2PKDL) and all of these were found to be similar to L. donovani isolates of 1970s epidemic.

Mechanism of action of an eukaryotic initiation factor-2 (eIF-2) associated 67 kDa glycoprotein (p67) and an eIF-2 kinase (dsI).

Mechanism of regulation of eIF-2 alpha-subunit phosphorylation by dsI and p67 was studied. The results are as follows: (1) At low dsI concentration, p67 protected equimolar concentration of eIF-2. (2) At high dsI concentration, dsI efficiently phosphorylated eIF-2 alpha-subunit even when equimolar concentrations of both p67 and eIF-2 were present. Significantly increased p67 concentration was necessary to protect eIF-2 alpha-subunit at high dsI concentration. (3) dsI was also phosphorylated as it phosphorylated eIF-2 alpha-subunit. p67 inhibited both eIF-2 alpha-subunit and dsI phosphorylation similarly. (4) Although the [32P]-labelled dsI formed during the reaction could be effectively chased upon subsequent addition of excess unlabelled eIF-2 and ATP, the [32P] labelled eIF-2 formed under identical conditions, retained most of the radioactivity. (5) dsI coimmunoprecipitated with three subunit eIF-2 and p67 inhibited this coimmunoprecipitation reaction. It has been proposed: Three subunit eIF-2 and free p67 are in equilibrium with eIF-2 bound to p67 and, eIF-2.p67 complex is resistant to dsI phosphorylation. Activated dsI is already phosphorylated. At high concentration, dsI(P) can bind to free three subunit eIF-2 and form eIF-2.dsI(P) complex. dsI(P) in this complex then transfers its phosphoryl residue to eIF-2 and forms eIF-2 alpha(P) in an irreversible reaction. In a subsequent reaction, unphosphorylated dsI is autophosphorylated using [gamma 32P]-ATP and the cycle continues. Inhibition of eIF-2 alpha-subunit phosphorylation by p67 blocks this phosphorylation cycle and consequent dsI phosphorylation.

Role of human placental extract on succinic dehydrogenase activity in carrageenin-induced edema in rats in vivo and its effect on erythrocyte lysis, platelet aggregation and trypsin activity in vitro.

Significant increase of liver succinic dehydrogenase (SDH, EC 1.3.99.1) activity was produced by carrageenin-induced edema in rats. Pretreatment with human placental extract inhibited the increased liver SDH activity in a dose-dependent manner. Placental extract was found to have little or no effect on the liver SDH activity in normal rats. Furthermore, heat-induced erythrocyte lysis was inhibited to a substantial extent by the extract and was found to be dose-responsive. However, adenosine diphosphate (ADP)-induced platelet aggregation and trypsin activity were not changed by the placental extract in vitro. The study indicates that the membrane stabilization and depletion of adenosine triphosphate (ATP) synthesis may contribute to antiinflammatory effect of the extract.

Career aspirations of dental students in a dental college in Calcutta, India.

Career aspirations of dental students at Dr. R. Ahmed Dental College & Hospital, Calcutta have been studied through a survey by questionnaire among 215 undergraduate dental students. The response rate was 78.6%. Private practice and clinical jobs were the commonest choices. Academic research jobs formed a very low priority area for career persual. Trends in junior through senior classes show low research interests in the senior as compared to the junior classes. More females preferred clinical jobs with private practice the second most common choice.

Role of nicotinic acid as modulator of liposomal microviscosity.

Using the fluorescent probe 1,6-diphenyl-1,3,5 hexatriene, we have investigated the effect of nicotinic acid, a derivative of the toxic alkaloid nicotine, on the fluidity profile and activation energy of diffusion in the liposomal system of several lipids. We have also studied how the fluidizing property of nicotinic acid affects the intermediate fluid condition induced by cholesterol in these liposomal systems.