Clinico-Pathological Profile of Basaloid Skin Tumors-Experience from a Tertiary Care Center of Eastern India.

Basaloid tumors comprise a wide spectrum of benign and malignant tumors like basal cell carcinoma, seborrheic keratosis, pilomatricoma, basosquamous carcinoma, trichoblastoma, and cylindroma. Among them, basal cell carcinoma is the most common type which constitutes about 90% of all malignant skin tumor. This study was aimed at analyzing the clinico-pathological profile of basaloid skin tumors attending radiotherapy and surgery OPD of our institution and compares them with those of the reported literature from rest of the country as well as outside world. All cases of basaloid skin tumors presented at radiotherapy, surgery, and dermatology OPD between January 2020 and June 2021 with or without a histological diagnosis were evaluated. Those without a histological diagnosis underwent biopsy and categorized according to standard histological criteria. After histological confirmation, we collected demographic, clinical, and pathological data of the cases. Among 106 patients analyzed, 54.7% (58) cases were diagnosed as basal cell carcinoma followed by seborrheic keratosis (17.9%), pilomatricoma (13.2%), basosquamous carcinoma (9.4%), trichoblastoma (2.8%), and cylindroma (1.8%). Mean age of presentation was 57.03 (± 7.435) years, and head-neck region was the most common site of involvement for basal cell carcinoma. Twenty-two cases required immunohistochemical assessment for confirmation of diagnosis. To conclude, this study is one of its first from Eastern India and will act as a stepping stone for future studies concentrating on clinico-pathological profile, early diagnosis and treatment of basaloid skin tumors.

Clinicopathological study and immunohistochemical analysis of expression of annexin A5 and apelin in human placentae of gestational diabetes mellitus.

BACKGROUND AND AIMS: Gestational diabetes mellitus (GDM) is one of the commonest medical complications of pregnancy. Annexin A5 (ANXA5) is a protein, found in apical surfaces of syncytiotrophoblasts, which prevents fetal and placental vascular thrombosis in GDM. Apelin is a bioactive peptide which has been linked to GDM. The aim of the present study was to correlate macroscopic as well as microscopic changes and immunohistochemical expression of ANXA5 and apelin in placentae of GDM with maternal and neonatal clinical features and also to compare the results with those in matched controls. METHODS: This prospective observational study was undertaken for a period of one year from April 2020 to March 2021. It comprised of 42 patients of GDM. Gross features, microscopic features and intensity and grade of expression of ANXA5 and Apelin were analyzed in placentae of GDM. RESULTS: Morphological changes detected in GDM placentae included increased immature villi (16 cases, 38%), increased syncytial knots (36, 86%), perivillous fibrin deposition (20, 48%), fibrosis of villous stroma (20, 48%), presence of nucleated red blood cells (12, 28.5%) and hypervascularity (34, 81%). The extent of histopathological changes noted in GDM placentae was significantly higher than that in matched controls. GDM placentae showed significantly reduced expression of ANXA5 and Apelin in terms of grade and intensity when compared with matched controls. Reduced expression (mild intensity) of ANXA5 was noted in 22 GDM cases (52.3%) whereas apelin expression was of weak intensity in 26 (61.9%) cases. Among GDM patients, statistically significant association was noted between ANXA5 intensity and neonatal resuscitation, apelin grade and preterm birth as well as low birth weight and apelin intensity and requirement of treatment in sick neonatal care unit. CONCLUSION: The placental expression of the proteins, ANXA5 and Apelin, is altered in GDM though their exact pathogenetic mechanisms are yet to be understood. They can be targets for development of prophylactic and therapeutic agents in future.

Drug Repurposing: Hydroxyurea Therapy Improves the Transfusion-Free Interval in HbE/Beta-Thalassemia-Major Patients with the XmnI Polymorphism.

Aims: HbE/ß-thalassemia is the most prevalent form of severe ß-thalassemia in Asian countries. Hydroxyurea (HU) is the most common drug used for the management of sickle-cell anemia but not thalassemia. In this study, we aimed to assess clinical HU response among the Bengali HbE/ß-thalassemia patients with respect to the XmnI γGglobin polymorphism and elucidate the association between this polymorphism and HU response efficacy. Materials and Methods: We enrolled 49 transfusion-dependent patients with HbE/ß-thalassemia. Fetal hemoglobin levels were measured using high-performance liquid chromatography and complete blood counts were determined pre- and post-HU therapy. Polymerase chain reaction-restriction fragment length polymorphism analyses were performed for genotyping the XmnI γGglobin polymorphism. Results: A total of 30 (61.22%) patients were found to be responders, whereas the remaining 19 (38.78%) were nonresponders. We found 33 patients with the heterozygous (C/T) and three with the homozygous mutant (T/T) genotype status. We obtained a statistically significant correlation (p < 0.001) between the XmnI polymorphism genotype and transfusion-free interval. Patients with the XmnI polymorphism were found to be good responders for HU therapy and showed increased hemoglobin levels. Conclusions: Our findings indicate that HU is a potential drug candidate for thalassemia management, particularly for HbE/ß-thalassemia. These results hold implications in repurposing HU as an effective and efficient therapy for HbE/ß-thalassemia.

Clinicopathological study of annexin A5 & apelin in pre-eclamptic placentae with emphasis on foetal outcome.

Background & objectives: Pre-eclampsia has remained an elusive disease with serious impacts on both maternal and foetal health. Two novel markers, annexin A5 (ANXA5) and apelin are currently of considerable interest. The present study aimed to determine the placental expression of ANXA5 and apelin in pre-eclamptic placentae and also to elucidate if there is any correlation between the expression of these markers and the clinical features of both, mother and neonate. The comparison between gross and histopathological features of pre-eclamptic placentae and controls was another objective. Methods: A prospective, observational study was undertaken for one year. Placentae of pre-eclamptic patients and matched controls (matched for age, ethnic and socio-economic background) were collected along with the clinical data. Gross and histopathological analyses were done and immunohistochemical study of placental sections with ANXA5 and apelin was also undertaken. Results: 79 pre-eclamptic patients and equal numbers of matched controls were included in the study. The difference in weight and dimensions of placentae between the pre-eclampsia group and matched controls was significant. Histopathological features noted in the pre-eclamptic placentae included decidual vasculopathy, infarction, perivillous fibrin deposition, increased syncytial knots and distal villous hypoplasia. There was a significant reduction in the expression of both ANXA5 and apelin in pre-eclamptic placentae compared to controls. Among pre-eclamptic patients, the intensity of ANXA5 and apelin expression showed a significant association with respect to neonatal resuscitation. Furthermore, the intensity of apelin showed expression a significant correlation with the requirement of sick neonatal care unit treatment. Interpretation & conclusions: The results of the present study suggest that both ANXA5 and apelin levels are reduced in pre-eclamptic placentae. Hence, it is recommended to further explore the impact of these markers on pregnancy outcomes by undertaking randomized controlled trials.

A study of the expression and localization of toll-like receptors 2 and 9 in different grades of cervical intraepithelial neoplasia and squamous cell carcinoma.

TLRs are important molecules of innate immune response, those play central role in host pathogen interaction and recognition through pathogen associated molecular patterns (PAMPs). Previous studies have indicated the role of TLRs in many human malignancy and cervical cancer in terms of viral recognition and inflammatory changes in-vivo. The objective of this study was to evaluate the expression and localization of toll-like receptor (TLR) 2 and TLR9 in preinvasive and invasive cervical cancer patients and to investigate its use as a probable diagnostic tool for better management cervical cancer. This single institution study includes individuals with normal, precancerous lesions, cervical intraepithelial neoplastic (CIN) and invasive squamous cell carcinoma (SCC) of the cervix. Upon confirmation by histopathology, fluorescence based immunohistochemistry was performed in all patients for TLR2 and TLR9, followed by semi-quantitative estimation of the staining intensity and grade of expression. The expression pattern of TLR2 and TLR9 does not vary greatly from normal to precancerous lesions, but a significant variation was observed in advance stages, i.e. squamous cell carcinoma of the uterine cervix. Additionally the expression increased marginally in higher grades. In spite of their low difference in expression along different stages of cervical cancer, both TLR2 and TLR9 could detect the disease at an advance stages as depicted by the receiver operator characteristics curve analysis.

Diversity and antifungal resistance patterns of prevalent opportunistic pathogenic yeasts colonizing the oral cavities of asymptomatic human immunodeficiency virus-infected individuals, and their relation to CD4(+) counts.

BACKGROUND: Yeasts are important opportunistic pathogens, in individuals infected with human immunodeficiency virus (HIV). Yeast species inhabiting the oral mucosa of HIV-infected persons can act as source of oral lesions, especially as the individual progresses towards immunocompromised state. Present study was conducted to evaluate the diversity of yeasts in oral cavities of asymptomatic HIV-infected persons and their association with CD4(+) cell counts. MATERIALS AND METHODS: 100 HIV seropositive subjects and 100 healthy controls were screened for oral yeast carriage using standard procedures. RESULTS: Of the 100 HIV-seropositive persons screened, 48 were colonized by different yeasts, either alone or in association with another species. Candida albicans was the most common species (56.90%) while non C. albicans Candida (NCAC) accounted for 39.65%. Among NCAC, Candida tropicalis and Candida krusei were most common. One isolate each of rare opportunistic pathogenic yeasts, Geotrichum candidum and Saccharomyces cereviseae, was recovered. The control group had an oral candidal carriage rate of 23%; C. albicans was the predominant species, followed by Candida glabrata, C. tropicalis and Candida parapsilosis. Antifungal susceptibility testing revealed no resistance in C. albicans, to the commonly used antifungal agents, whereas resistance or dose dependent susceptibility to fluconazole was observed in some of the NCAC species. CONCLUSION: Oral carriage of opportunistic pathogenic yeasts was greater in HIV-seropositive persons heading towards immunocompromised state, as evidenced by their CD4(+) cell count. The predominant yeast isolated in this study (C. albicans), was found to be susceptible to commonly used antifungals.

Expression of matrix metalloproteinase-2 and 9 in cervical intraepithelial neoplasia and cervical carcinoma among different age groups of premenopausal and postmenopausal women.

PURPOSE: The objective was to study the gelatinolytic activity of matrix metalloproteinase-2 (MMP-2) and matrix metalloproteinase-9 (MMP-9) in preinvasive and invasive carcinoma of the uterine cervix. The expressions were analysed against different age groups, as to demonstrate whether the expression of MMP-2 and MMP-9 is an early or a late event during the progression of cervical cancer. Additionally, the diagnostic accuracy of MMP-2 and MMP-9 was evaluated with ROC curve. METHODS: A total number of 180 samples of cervical tissue were studied for MMP-2 and MMP-9 gelatinolytic activity. The cases were selected as to include 63 normal cases, 94 CIN cases and 23 cervical carcinoma cases. Among 94 CIN cases, 40 were CIN1, 26 were CIN2 and 28 were CIN3, as reported by histopathology. The gelatinolytic activities of MMP-2 and MMP-9 were evaluated by gelatin zymography in premenopausal and postmenopausal groups. RESULTS: MMP-2 expressions (latent and active) were very low in control samples, followed by increase in CIN1, decrease in CIN2 and further increase in advance stages. MMP-9 had also shown the same expression pattern that of MMP-2. While comparing the expression of MMP-2 and MMP-9 in different age groups, we found initial CIN stages were prevalent in early age that expressed considerable amount of MMP-2 and MMP-9, and advance stages of carcinoma cervix were prevalent at an elderly age. CONCLUSION: Both MMP-2 and MMP-9 have role in cancer progression and remodelling of the ectocervix. Although expression level varies intricately, a distinctive ROC curve demonstrated MMP-2 active form and MMP-9 form could be used in diagnostic purpose in detection of cervical lesion and cancer.

Skull base chordoma presenting as nasopharyngeal mass with lymph node metastasis.

Spheno-occipital chordomas can rarely present as nasopharyngeal mass. Metastases occur only in advanced disease. They can pose a diagnostic dilemma when information about diagnosis of the primary tumor is not available. We present cytological findings in upper cervical lymph node of a case of nasopharyngeal chordoma and discuss possible differential in such a location.

Collecting duct carcinoma: a rare malignancy.

A 45-years old man complained of hematuria, and subsequent examination and ultrasonography revealed a mass in the left kidney. Nephrectomy was performed and macroscopically an ill-defined pale-cream, irregular mass was identified which occupied predominantly the renal medulla. Histopathologic examination showed slit like tubular ducts lined by atypical cuboidal to polygonal cells and a marked desmoplastic stromal reaction. The diagnosis of collecting duct carcinoma was made. Patient is now doing well after 11 months of follow up.

Cytokeratin 19 immunoreactivity in the diagnosis of papillary thyroid carcinoma.

CONTEXT: The diagnosis of papillary thyroid carcinoma (PTC) is based on nuclear features. These features may be present in focal areas in benign thyroid diseases and follicular adenoma (FA), leading to diagnostic difficulty. AIMS: To evaluate the expression and pattern of the distribution of cytokeratin 19 (CK19) in PTC and compare its reactivity with other neoplastic and non-neoplastic conditions to assess its potential as a useful marker for PTC. MATERIALS AND METHODS: Twenty two cases of papillary carcinoma (usual type, follicular and diffuse sclerosing variant), eight follicular adenomas, eight multinodular goiters (MNG) were collected for a period of two years and six months. Sections were taken from thyroidectomy specimens fixed in 10% buffered neutral formalin. Hematoxylin and eosin staining and immunohistochemical staining for CK19 were done using standard protocol. Results were semiquantitatively scored as follows: 1+ (<5% positively stained cells), 2+ (5-25%), 3+ (25-75%) and 4+ (>75%), and then analyzed. STATISTICAL ANALYSIS AND RESULTS: All 22 (100%) papillary carcinomas showed diffuse and strong (3+ and 4+) CK19 expression. Six out of eight (75%) FAs and four out of eight (50%) MNG were positive for CK19, but it was of weaker intensity (1+ and 2+) and focal in distribution. CONCLUSION: Focal CK19 staining may be found in benign disease, but diffuse and strong positivity is characteristic of PTC, which can be used in the diagnosis of PTC in lesions of equivocal morphological appearances.

A study of histopathological spectrum and expression of Ki-67, TP53 in primary brain tumors of pediatric age group.

OBJECTIVES: The primary brain tumors are the second most common cause of death due to malignancies in children. This study was done to analyze the histological spectrum of primary brain tumors in children and also to find out the expression of p53 and Ki67 in some of the common pediatric brain tumors. MATERIALS AND METHODS: This study was done over a period of 2.5 years. The patients were followed up until 6 months to determine the outcome. We examined H and E sections from 61 pediatric brain tumors and also performed immunohistochemical stains with p53 and Ki67 on 52 of these samples. RESULTS: Of the 61 cases of pediatric brain tumors the commonest were pilocytic astrocytomas and medulloblastomas both constituting 22.9% of total cases, followed by high grade gliomas, that is, anaplastic astrocytoma and glioblastoma taken together (14.7%), diffuse astrocytomas (11.4%), ependymomas (8.1%), and oligodendrogliomas (4.9%). Other cases comprised craniopharyngiomas, astroblastomas, and gangliocytoma. The mean age of presentation was 9.3 years, male children being more commonly affected. Ki67 labeling index (LI) and p53 expression in pilocytic astrocytomas and diffuse astrocytomas were significantly lower than that of high-grade astrocytomas. However, there was no significant difference of expression of these two antigens in pilocytic astrocytomas and diffuse astrocytomas. It was found that Ki67 LI was a better marker for distinguishing between grades of astrocytoma than p53 (P=0.000 and P=0.002, respectively). The survival in cases of pilocytic astrocytomas was far better than high-grade gliomas. However, there was no significant difference in survival between pilocytic astrocytoma and diffuse infiltrating astrocytoma. There was significant positive correlation between expression of p53 and Ki67 LI in cases of medulloblastomas. Both p53 (P=0.002) and Ki67 LI (P=0.000) taken individually correlated well with survival in these cases. Also, Ki67 LI is better predictor of outcome than p53. CONCLUSION: From this study, it can be concluded that Ki67 and p53 score correlated well with the grade of astrocytoma; however, Ki67 is a better marker for differentiating between the grades of astrocytoma than p53. Also, Ki67 LI is a better prognostic factor than p53 in case of medulloblastomas.

Screening for thalassemia and other hemoglobinopathies in a tertiary care hospital of West Bengal: implications for population screening.

Hemoglobinopathies are common genetic disorders of hemoglobin, which can be prevented by population screening and offering genetic counseling. In absence of population-based screening for hemoglobinopathies, the hospital-based diagnosis register provide idea about the extent of problem in the community. The present study was undertaken to find out the burden of hemoglobinopathies and spectrum of this disorders among the population who were screened in the hospital-based screening program. A record-basedanalysis of subjects who underwent screening for hemoglobinopathies in Burdwan Medical College and Hospital over a period of 3 years and 4 months revealed that overall 29.3% of subjects were positive for hemoglobinopathies. Beta thalassemia heterozygous was the most commonhemoglobinopathy in this region closely followed by hemoglobin E heterozygous. In view of high prevalence of hemoglobinopathies in this region, a routine premarital screening program is needed for identification and prevention of high-risk marriages.

Molecular typing and in vitro fluconazole susceptibility of Candida species isolated from diabetic and nondiabetic women with vulvovaginal candidiasis in India.

BACKGROUND: Candida albicans and Candida glabrata are the major causes of vulvovaginal candidiasis (VVC) in Indian women with diabetes mellitus. Little information is available regarding the genotyping of Candida species isolated from Indian diabetic women with VVC. METHODS: In this study, a total of 57 Candida species, comprising Candida albicans and Candida glabrata, isolated from Indian women with VVC, were genotyped and tested for fluconazole susceptibility. Arbitrarily primed polymerase chain reaction (AP-PCR) was used to genotype C glabrata isolates, whereas Southern blot hybridization using a Candida albicans repetitive element-2 (CARE-2) probe was used to genotype C albicans. RESULTS: Genotyping showed that all the C albicans isolates were genetically heterogenous. The pattern of DNA bands obtained after AP-PCR for C glabrata strains were predominantly conformed to genotype A. In vitro fluconazole-susceptibility testing of the isolates using the Clinical and Laboratory Standards Institute M27A2 protocol showed that more than 93% of the Candida isolates were susceptible. CONCLUSIONS: Ninety-five percent of the C albicans isolates analyzed were different and genetically unrelated. The analysis of the AP-PCR DNA banding pattern of C glabrata isolates showed that it resembled genotype "A". The Candida isolates were found to be susceptible to fluconazole, with minimum inhibitory concentrations ranging from 0.5 µg/mL to 8 µg/mL. This correlates with the use of fluconazole as a first-choice antifungal for treating VVC in India.

Current scenario of cryptococcosis and antifungal susceptibility pattern in India: a cause for reappraisal.

This study analysed the spectrum, antifungal susceptibility pattern, clinical course and molecular epidemiology of cryptococcosis. Four hundred and thirty-nine samples obtained from 378 meningitis patients were processed by standard procedures. Minimum inhibitory concentration (MIC) of fluconazole and amphotericin B for the isolates was tested by broth micro dilution and by E-strip method. Molecular analysis by random amplified polymorphic DNA-PCR of eight isolates was performed using M13 primer. Cryptococcosis was diagnosed in 35 patients [HIV-1 seropositive (19) and apparently immunocompetent (16)]. Cryptococcus neoformans var. neoformans (serotype A and D) was the predominant isolate on phenotypic identification. Three C. neoformans var. gattii were isolated from HIV-1 seropositive (2) and apparently immunocompetent (1) patients. MIC 90 for amphotericin B and fluconazole were 1 and 8 mug ml(-1) respectively. On RAPD-PCR, less diversity was seen among Indian isolates. AIDS remains the single most important risk factor for cryptococcosis. Rising MIC of the available induction and maintenance drugs is of grave concern. The DNA typing technique showed less diversity among Indian strains. Routine surveillance and application of molecular typing methods are crucial to know the baseline and existing pattern of cryptococcosis.

Differences in the cell wall architecture of melanin lacking and melanin producing Cryptococcus neoformans clinical isolates from India: an electron microscopic study

Cryptococcus neoformans is an important opportunistic fungal pathogen that causes life-threatening infection of the central nervous system. A major virulence factor for C. neoformans is the production of melanin in the cell wall. Using transmission electron microscopy, we studied the cell walls of three pairs of isolates obtained from patients with dual cryptococcal infections, where a melanotic and an albino strain were isolated from the CSF of each patient. Transmission Electron Microscopy revealed that the albino strains lacked a melanin layer whereas a melanin layer was associated with the cell wall of the melanotic strains, comprising approximately 75 percent of the cell wall area. The cell wall size of the melanin producing cells was approximately double the size the albino isolates' cell walls (p value <= 0.003). In this study TEM revealed that the differences in the ultrastructure of the melanin lacking and melanin producing isolates were associated to the cell wall and the melanin layer.

Cryptococcus neoformans é um importante fungo oportunista patogênico que causa infecção no sistema nervoso central, e que pode levar o paciente à morte. Um dos principais fatores de virulência do C. neoformans é a produção de melanina na parede celular. Utilizando microscopia eletrônica de transmissão, nós estudamos as paredes celulares de três pares de isolados obtidos de pacientes com dupla infecção pelo fungo, onde um isolado melanizado e um albino foram isolados do líquor de cada paciente. A microscopia eletrônica de transmissão revelou que as cepas albinas não apresentavam a camada de melanina enquanto que uma camada de melanina estava associada com a parede celular de cepas melanóticas, constituindo aproximadamente 75 por cento da área da parede celular. O tamanho da parede celular das células produtoras de melanina foi aproximadamente o dobro do tamanho da parede celular dos isolados albinos (p < 0,003). Neste estudo, a microscopia eletrônica de transmissão revelou que as diferenças na estrutura dos isolados albinos sem melanina e dos isolados produtores de melanina estava associada à parede celular e a camada de melanina.

Prolonged (3-month) mycological cure rate after boric acid suppositories in diabetic women with vulvovaginal candidiasis.

OBJECTIVE: Patients with diabetes mellitus (DM) are at increased risk of vulvovaginal candidiasis (VVC) due to C. glabrata. In our previous study we had shown that patients with diabetes mellitus and VVC show an overall superior mycological cure rate (74% versus 51%) with boric acid therapy at 15th day as compared to fluconazole. Present study was carried out to assess long term response to boric acid in diabetic women with VVC. MATERIAL AND METHODS: Subjects included 40 consecutive diabetic women (type 2 DM=26 and type 1 DM=14) who had achieved mycological cure (high vaginal swab culture negativity) on day 15 of therapy following single-dose oral-150 mg fluconazole (n=21) or 600 mg of boric acid suppositories given daily for 14 days (n=19). At third month of follow up, patients were assessed for signs and symptoms of VVC and a repeat HVS was collected for fungal culture. HbA1c was measured to assess glycaemic control. RESULTS: The mean age, BMI, HBA1c and frequency of various Candida species isolated at initial diagnosis were comparable in the fluconazole and boric acid treatment groups. Fifteen of 21 (71.4%) and 12 of 19 (63.1%) women who achieved mycological cure at 15 day remain cured at three months in the fluconazole and boric acid treated groups, respectively (P=0.83). With 74% mycological cure at 15th day, this would indicate that on an average only 46.6% of diabetic women with VVC would remain cured at 3 months after a course of 14 days boric acid therapy. Most of the patients relapsed with no change in Candida species. The demographic profile and mean HbA1c (8.6+/-2.2 versus 8.8+/-2.4%, P=0.83) were comparable in patients with (n=27) and without mycological cure (n=13). CONCLUSION: The results of the current study indicating comparable mycological cure rate at 3 months between fluconazole and boric acid treated patients would support use of boric acid in the acute management of VVC in view of its superior short term response in diabetic women with C. glabrata infections. However, there is need to explore other therapeutic regimens which are effective in achieving long term mycological cure in diabetic women with VVC.

Prevalence of Candida glabrata and its response to boric acid vaginal suppositories in comparison with oral fluconazole in patients with diabetes and vulvovaginal candidiasis.

OBJECTIVE: A large proportion of vulvovaginal candidiasis (VVC) in diabetes is due to non-albicans Candida species such as C. glabrata and C. tropicalis. Observational studies indicate that diabetic patients with C. glabrata VVC respond poorly to azole drugs. We evaluated the response to oral fluconazole and boric acid vaginal suppositories in diabetic patients with VVC. RESEARCH DESIGN AND METHODS: A total of 112 consecutive diabetic patients with VVC were block randomized to receive either single-dose oral 150-mg fluconazole or boric acid vaginal suppositories (600 mg/day for 14 days). The primary efficacy outcome was the mycological cure in patients with C. glabrata VVC in the two treatment arms. The secondary outcomes were the mycological cure in C. albicans VVC, overall mycological cure irrespective of the type of Candida species, frequencies of yeast on direct microscopy, and clinical symptoms and signs of VVC on the 15th day of treatment. Intention-to-treat (ITT; n = 111) and per-protocol (PP; n = 99) analyses were performed. RESULTS: C. glabrata was isolated in 68 (61.3%) and C. albicans in 32 (28.8%) of 111 subjects. Patients with C. glabrata VVC showed higher mycological cure with boric acid compared with fluconazole in the ITT (21 of 33, 63.6% vs. 10 of 35, 28.6%; P = 0.01) and PP analyses (21 of 29, 72.4% vs. 10 of 30, 33.3%; P = 0.01). The secondary efficacy outcomes were not significantly different in the two treatment arms in the ITT and PP analyses. CONCLUSIONS: Diabetic women with C. glabrata VVC show higher mycological cure with boric acid vaginal suppositories given for 14 days in comparison with single-dose oral 150-mg fluconazole.

The glyoxylate cycle enzyme activities in the pathogenic isolates of Candida albicans obtained from HIV/AIDS, diabetic and burn patients.

Recently it has been found that Candida albicans harbours enzymes involved in the glyoxylate cycle (GC), which have a role in its virulence, especially the two key enzymes, isocitrate lyase (ICL) and malate synthase (MS). There are however, few studies on the GC enzyme activities isolated in the clinical isolates. Samples were collected from three groups of patients namely, HIV/AIDS, diabetic and burn patients suffering from candidiasis at different body locations. Isolation, identification and the antifungal susceptibility test of all the isolates of C. albicans were followed by the standard techniques. Measurements of all the GC enzyme activities were also carried out by the standard methods. Levels of the principal GC enzymes showed significant changes when calculated and compared taking control strains of C. albicans. The activity of the two key enzymes of the GC, ICL and MS were significantly higher in the isolates from diabetic patients. No significant relationship between the drug susceptibility and the level of enzymes of the GC was observed. As GC activity is absent in mammalian cells, a specific inhibitor for the GC could be developed and these enzymes therefore can be used as a new antifungal target.

Pattern of Candida species isolated from patients with diabetes mellitus and vulvovaginal candidiasis and their response to single dose oral fluconazole therapy.

OBJECTIVE: Patients with diabetes mellitus are at increased risk of vulvovaginal candidiasis (VVC). Besides Candida albicans, they often have infection due to non-C. albicans Candida species such as C. glabrata. Oral single dose fluconazole (150 mg) is commonly used to treat VVC in non-diabetic individuals with response rate varying from 70 to 90%. However, there is paucity of related information in diabetic women with VVC. Present study has been conducted to systematically assess the effect of fluconazole therapy among diabetic patients with clinically symptomatic VVC. METHODS: Study subjects included 85 consecutive patients with diabetes mellitus (type 2=70 and type 1=15) and 62 non-diabetic women who had clinical signs and symptoms of VVC and in whom evidence of candidiasis was documented by presence of yeast on direct microscopy followed by culture. Single dose fluconazole (150 mg) was given orally to all the subjects in a supervised manner. Subjects were reassessed on 14th day after fluconazole therapy and a repeat high vaginal swab was taken for direct microscopy and fungal culture. Total glycosylated haemoglobin (HbA1) was measured to assess glycaemic control. RESULTS: There were no significant differences in the frequency of pruritus (55.9 vs. 56.7%), vaginal discharge (63.8 vs. 69.0%), dyspareunia (25.0 vs. 20.0%), and percentage yeast positivity (67.5 vs. 54.7%) between diabetic and control groups before the start of fluconazole therapy. Following fluconazole therapy, vaginal discharge on examination and yeast positivity on direct microscopy continued to remain positive in higher percentage of subjects in the diabetic group as compared to non-diabetic subjects (52.5 vs. 36.4%; P =0.22 and 50.7 and 29.0%, respectively, P =0.07, respectively). Overall 67.1% of patients with diabetes and 47.3% of controls continued to show persistence of Candida growth on high vaginal swab culture following fluconazole treatment (P=0.042). Candida glabtara was the most common species isolated in patients with diabetes mellitus and its frequency was significantly higher in them when compared to control group (54.1 vs. 22.6%, P<0.001). C. albicans was the most common species isolated in controls. Species-specific response to fluconazole showed that 81.3% of patients in the diabetic group and 78.6% of the non-diabetic controls continued to show fungal growth when C. glabrata was the organism grown (P=0.99). However, in case of C. albicans, 45.4% of the patients in the diabetic group and only 21.5% of the controls had persistent Candida growth following fluconazole therapy (P=0.22). CONCLUSION: Overall only one third of patients with diabetes mellitus and VVC respond to single dose 150 mg of fluconozole therapy. Limited response in the clinical symptoms and culture negativity following single dose fluconazole therapy in diabetic subjects with VVC is explained by the high prevalence of C. glabrata in them. The present study involved only 85 patients and majority of them had type-2 diabetes mellitus. There is need to perform similar study in large number of diabetics subjects including patients with type-1 diabetes mellitus and assess various alternative treatment protocol which are also effective in C. glabrata infection.

Dual infections with pigmented and albino strains of Cryptococcus neoformans in patients with or without human immunodeficiency virus infection in India.

Cryptococcus neoformans is an encapsulated yeast-like fungus of worldwide distribution. Melanin production is an important virulence factor of C. neoformans. We report the identification of distinct cryptococcal isolates with either pigmented or white colony phenotypes on L-dihydroxyphenylalanine agar plates in three patients who presented with meningitis to the All India Institute of Medical Sciences in India. Two of the patients were also infected with human immunodeficiency virus. Biochemical studies, India ink analysis, immunofluorescence with antibodies specific to capsular antigen, and serotyping confirmed that the melanotic and albino strains were C. neoformans serotypes A and D, respectively. Genotyping with M13 and [GACA]4 primers revealed that all the C. neoformans isolates were genetically different. The CNLAC1 gene associated with melanin production was identified in all the strains by PCR. Standard MIC testing revealed that the strains had similar susceptibilities to amphotericin B, but time-kill assays with the antifungal showed reduced susceptibility in melanin-producing strains. Infection studies with A/Jcr mice showed that the melanin-lacking yeast were less virulent than melanin-producing isolates. These findings indicate that these patients had dual infections with pigmented and albino strains of C. neoformans that were phenotypically and biologically different. Continued surveillance of primary isolates from patients with cryptococcosis by analyzing phenotypic differences and by molecular methods may reveal that mixed infections occur more commonly than is currently realized.