Environmental enrichment preserves hippocampal neurons in diabetes and stressed rats.

This study evaluated the effect of Environmental Enrichment (EE) on neuron morphology in the CA1, CA3 and dentate hilus (DH) regions of the hippocampus by quantitating the total dendritic arborizations. EE is a potential intervention for stress and diabetes. It is capable of mitigating diabetes and stress-induced cognitive and memory deficit. Diabetes and stress were induced in male Wistar rats (4-5 weeks). Diabetic and stressed rats were exposed to EE on Day 2 post STZ injection and subsequently once daily for 30 days. All animals were sacrificed on Day 30. The hippocampus was dissected and processed for Golgi staining to quantitate dendritic arborizations at the CA1, CA2 and DH regions. Diabetes (D) and Diabetes+stress (D+S) groups had significantly fewer apical and basal dendritic branching points (ADBP, BDBP) at CA1 (p<0.01), CA3 (p<0.001) and DH (p<0.001) relative to control group (NC). Diabetes and stressed rats exposed to EE: [D+EE and D+S+EE groups] exhibited significantly denser ADBP and BDBP at all regions relative to D (p<0.001) and (D+S+EE) (p<0.001) groups respectively. EE significantly preserved neuronal arborizations in hippocampus of diabetic and stressed rats, suggesting a potential entity of diabetes and stress management.

Contribution of macrophages to fetomaternal immunological tolerance.

The semi-allogeneic fetus develops in a uniquely immune tolerant environment within the uterus. For successful pregnancy, both the innate and adaptive immune systems must favor acceptance of the fetal allograft. Macrophages are the second most abundant immune cells after natural killer (NK) cells in the decidua. In coordination with decidual NK cells and dendritic cells, macrophages aid in implantation, vascular remodeling, placental development, immune tolerance to placental cells, and maintenance of tissue homeostasis at the maternal-fetal interface. Decidual macrophages show the classical activated (M1) and alternatively activated (M2) phenotypes under the influence of the local milieu of growth factors and cytokines, and appropriate temporal regulation of the M1/M2 switch is vital for successful pregnancy. Disturbances in the mechanisms that control the M1/M2 balance and associated functions during pregnancy can trigger a spectrum of pregnancy complications ranging from preeclampsia and fetal growth restriction to preterm delivery. This review addresses various mechanisms of tolerance, focusing on the basic biology of macrophages, their plasticity and polarization, and their protective roles at the immune-privileged maternal-fetal interface, including direct and indirect roles in promoting fetomaternal immune tolerance.

Development and quality assessment of carrageenan incorporated low fat chevon patties.

The present study was conducted to develop and assess the quality of carrageenan incorporated chevon patties with the objective of reducing fat content. Efficacy of carrageenan as fat replacers (0.3, 0.6 & 0.9 %) was assessed for development of low fat chevon patties. Emulsion stability and cooking yield increased with the increase in levels of carrageenan. Significantly (P < 0.05) lower fat and cholesterol contents and higher moisture and fat retention were observed in formulation with carrageenan. No significant difference in the mineral content in either of the treatment was recorded. Incorporation of fat replacer in chevon patties demonstrated significant effect on all the textural parameters except adhesiveness. Results of color value illustrated that lightness (L*) value differ significantly. Sensory scores were higher or comparable for patties containing 0.6 % carrageenan as compared to control. Hence, carrageenan was observed to be suitable as fat replacer for producing low fat chevon meat patties.

Clinical utility of urine neutrophil gelatinase-associated lipocalin measured at admission to predict outcomes in heterogeneous population of critically ill patients.

Urine neutrophil gelatinase-associated lipocalin (uNGAL) is a reliable early biomarker of acute kidney injury (AKI) in a homogeneous patient population. However, its utility in a heterogeneous population of critically ill, in whom the time of onset of renal insult is often unclear, is not clearly established. We evaluated the ability of a single measurement of uNGAL in a heterogeneous adult population, on admission to intensive care unit (ICU), to predict the occurrence of AKI and hospital mortality. One hundred and two consecutive adult patients had uNGAL measured within 8 h of admission to ICU. The demographic and laboratory data were collected at admission. The diagnosis of AKI was based on AKI Network (AKIN) criteria. The primary outcome was the development of AKI, and the secondary outcome was hospital mortality. The mean age was 54 ± 16.4 years and 65% were males. Urine NGAL (ng/ml) was 69 ± 42 in patients with AKI (n = 42) and 30.4 ± 41.7 in those without AKI (P < 0.001). The area under the receiver operating characteristic (ROC) curve for prediction of AKI was 0.79 and for serum creatinine (SCr) was 0.88. The sensitivity and specificity for a cut-off value of uNGAL of 75 ng/ml to predict AKI were 0.5 and 0.85 respectively. uNGAL > 75 ng/ml was a strong (odd ratio = 5.17, 95% confidence interval: 1.39-19.3) and independent predictor of hospital mortality. A single measurement of uNGAL at admission to ICU exhibited good predictive ability for AKI though the sensitivity was low. The predictive ability of uNGAL was inferior to simultaneously measured SCr at admission, hence limited its clinical utility to predict AKI. However, admission uNGAL was a strong, independent predictor of hospital mortality.

Utility of fine needle aspiration cytology in evaluation of lymphadenopathy - An audit from a Cancer Centre in South India.

BACKGROUND: Cytological evaluation and diagnosis of lymphadenopathy plays an important role in distinction between reactive hyperplasia and malignancy.Being a tertiary cancer care centre, lymph node enlargement clinically suspected to be due to malignancy constitute the commonest indication for fine needle aspiration cytology (FNAC) at our centre. The aim of this study was to determine the utility of FNAC in evaluating enlarged lymph nodes and to categorizethe causes of lymphadenopathy diagnosed byFNAC at our centre. MATERIAL AND METHODS: Data was collected from the records of department of Pathology over a period of three months from January to March 2014. The data was analyzed and various parameters studied. RESULTS: There were 2000 aspirates over a period of three months of which 270 (13.5%) were from lymph nodes. Of these, 130 cases (48.2%) have metastatic deposits.We also came across 16 cases (5.9%) of lymphoma, 5 cases (1.9%) of granulomatous lymphadenitis, 2 cases (0.7%) of suppurative lesion and 99 cases (36.7%) of reactive hyperplasia during this period. In 3 cases (1.1%), the lesion turned out to be of salivary gland origin. Aspirates were suboptimal for diagnosis in 15 cases (5.5%). Aspirates were more in males (181) as compared to females (89). The most common site of aspiration was the cervical lymph node(64.5%),followed by supraclavicular=81(30%), inguinal=33(12.22%), axillary=19(7.04%) sub mental=1(0.37%) and others=7(2.59%). Apart from metastatic carcinomas other metastatic malignancies we came across were malignant melanoma, neuroblastoma, germ cell tumor and synovial sarcoma. CONCLUSION: FNAC of lymph nodes helps in rapid diagnosis of lymphadenopathy.Categorizingthe cause of lymph node enlargement as metastatic malignancy,lymphoma, reactive change, inflammatory cause,suppuration etc. can be done by FNAC.In patients with known histologically proven malignancy in whom a subsequent enlargement of lymph node occurs,a cytological diagnosis of metastasis helps in avoiding unwanted surgery for confirming metastasis.In patients without a previous diagnosis of malignancy, FNAC not only confirm metastatic deposit but in most conditions give a clue regarding site of primary. The use of immunocytochemistry and cell block preparations have increased the scope of FNAC.

The Revised Sonographic NASCET Index: A New Hemodynamic Parameter for the Assessment of Internal Carotid Artery Stenosis.

PURPOSE: A previously described Doppler parameter, the sonographic NASCET index (SNI), was derived to be more directly analogous to the North American Symptomatic Carotid Endarterectomy Trial (NASCET) methodology for assessing carotid artery stenosis. However, this index does not account for complex changes affecting the Doppler waveform. We propose a revised SNI (rSNI) in an effort to improve predicting carotid stenosis. MATERIALS AND METHODS: 25 carotid bifurcations with stenoses ranging from 40 - 92 % were analyzed. For each vessel, the rSNI and original SNI were calculated. The peak systolic velocity (PSV), rSNI, and original SNI were correlated with angiography using linear regression analysis and relative accuracies were compared at two thresholds. RESULTS: A correlation between rSNI and angiography was found to be significantly better than that between PSV or internal carotid artery-common carotid artery (ICA-CCA) peak velocity ratio and angiography (r² = 0.47 vs. 0.22; r² = 0.47 vs. 0.16). The accuracy of PSV in predicting high-grade stenosis was 68% and 72%, compared with 80% and 88% for rSNI, at each of two thresholds. The original SNI better correlated with angiography compared to the rSNI (r² = 0.55 vs. 0.47), but with slightly lower accuracy in predicting high-grade stenosis (76% vs. 80%). CONCLUSION: The revised SNI correlates more closely with angiographic stenosis than either the PSV or the ICA-CCA ratio, and is more accurate in predicting high-grade stenosis. However, it is overall comparable to the original SNI, suggesting that the previously unaccounted for effects over the remainder of the cardiac cycle do not significantly improve the ability to sonographically predict significant stenosis.

Nonlinear control of voltage source converters in AC-DC power system.

This paper presents the design of a robust nonlinear controller for a parallel AC-DC power system using a Lyapunov function-based sliding mode control (LYPSMC) strategy. The inputs for the proposed control scheme are the DC voltage and reactive power errors at the converter station and the active and reactive power errors at the inverter station of the voltage-source converter-based high voltage direct current transmission (VSC-HVDC) link. The stability and robust tracking of the system parameters are ensured by applying the Lyapunov direct method. Also the gains of the sliding mode control (SMC) are made adaptive using the stability conditions of the Lyapunov function. The proposed control strategy offers invariant stability to a class of systems having modeling uncertainties due to parameter changes and exogenous inputs. Comprehensive computer simulations are carried out to verify the proposed control scheme under several system disturbances like changes in short-circuit ratio, converter parametric changes, and faults on the converter and inverter buses for single generating system connected to the power grid in a single machine infinite-bus AC-DC network and also for a 3-machine two-area power system. Furthermore, a second order super twisting sliding mode control scheme has been presented in this paper that provides a higher degree of nonlinearity than the LYPSMC and damps faster the converter and inverter voltage and power oscillations.

A comparison of sagittal short T1 inversion recovery and T2-weighted FSE sequences for detection of multiple sclerosis spinal cord lesions.

PURPOSE: Multiple sclerosis (MS) is the most common disabling CNS disease of young adults. MRI is routinely used for the detection of MS plaques in the brain and spinal cord. A significant portion of patients with MS demonstrates spinal cord lesions at the time of initial workup, and these lesions are an important part of the McDonald criteria for diagnosis. However, whereas brain imaging sequences are now fairly standardized, there continues to be debate about the optimal sequences for imaging the spinal cord. The short T1 inversion recovery (STIR) sequence has been shown in the current literature to improve lesion detection with its additive T1/T2 weighting, but current spinal cord imaging protocols from the Consortium on MS Center Consensus Guidelines do not include the STIR sequence. We demonstrate that not only do STIR sequences improve lesion detection when compared directly with conventional T2-weighted sequences, but that they also significantly improve lesion conspicuity, facilitating earlier positive diagnosis and management. MATERIALS AND METHODS: Dedicated MR spinal cord imaging of twenty-nine sequential patients with clinically confirmed multiple sclerosis was retrospectively analyzed by two independent neuroradiologists in a novel study design. Sagittal T2-weighted and STIR sequence images from the same study for each patient were examined for MS plaques using a double-blinded review of individual images 'separated in time and space', such that STIR and T2 image pairs were never analyzed simultaneously. Number of lesions and lesion conspicuity for each lesion, using a subjective scale (1-5), were tallied for each sequence. Averages for each observer were compared using a paired t-test analysis for statistical significance, and assessment of inter-rater agreement was assessed using Cohen's kappa index. RESULTS: Significantly, more MS lesions were detected on STIR than on T2-weighted sequences for both observers (P = 0.001 and P = 0.005). In seven patients, the conventional T2 sequence detected no lesions at all, whereas STIR sequences showed significant cord involvement. Lesion conspicuity was also significantly better on STIR for both observers (P < 0.0005). This improved conspicuity leads to more uniform lesion detection. On the conventional T2-weighted sequence, there was a statistically significant difference in the number of lesions detected between the two observers (P = 0.003), but there was no statistically significant difference on STIR (P = 0.43). The kappa index showed greater interobserver agreement in both lesion count and lesion conspicuity on the STIR sequence as compared with T2. CONCLUSIONS: Short T1 inversion recovery sequence imaging not only significantly improves detection of MS lesions within the spinal cord, but also provides better contrast and conspicuity of visible lesions, creating a more confident diagnostic measure of MS extent and progression. Short T1 inversion recovery sequences of the spinal cord should be routinely obtained during initial and routine follow-up of MS.

Indian childhood cirrhosis (ICC) & ICC-like diseases: the changing scenario of facts versus notions.

Indian childhood cirrhosis (ICC), a disease considered to have been endemic in and unique to India has now been documented in children of non-Indian origin from other countries. More recently available findings from a large multicentre study sponsored by the Indian Council of Medical Research (ICMR) have dispelled some of the generally accepted notions and have established several new facts on different aspects of the disease. All relevant reports on ICC and ICC-like diseases, till date, were reviewed to obtain a proper perspective on the current state of our understanding on this non-Wilsonian copper overload liver disease. A primary role of exogenous copper in causing the disease was earlier debated on the basis of studies in India but investigators abroad studying some sporadic cases and a series of endemic ICC-like diseases supported a hepatotoxic injury by ingested copper in genetically susceptible infants and children in ICC- like disease and in ICC. Epidemiologic and morphologic findings in the well controlled ICMR study based on 225 cases of ICC and 426 controls, all confirmed on liver biopsy, have however, convincingly refuted this concept. Additionally, this study revealed that unlike what has been believed earlier, older children more than 3 yr age can get the disease and that in its natural course the hepatic histology can transform between the characteristic one considered diagnostic and some other patterns, any one of which can be the morphologic manifestation at first presentation of the patient. Older children and cases with milder morphologic changes at presentation had longer survival. The overall inference from critical analysis of all available data is that ICC and ICC-like diseases clinically manifest in a child of any age though common in younger ones, and a clinical diagnosis must be made in any child with so-called 'cryptogenic cirrhosis'. Exposure to exogenous copper in food, milk and water should not be a prerequisite for this consideration. A liver biopsy whenever feasible should be mandatory for confirmation with the understanding that the morphologic changes in liver can present a few other patterns besides the characteristic one currently taken to be diagnostic. The ascribed current decline in encountering ICC is likely to be due partly to missing a diagnosis and partly to a true reduction in incidence consequent on time related economic and socio-cultural changes.

Molecular & phenotypic characterization of Staphylococcus epidermidis in implant related infections.

BACKGROUND & OBJECTIVES: The discrimination between the Staphylococcus epidermidis colonizing the deep seated indwelling devices and those which are mere commensals has always been a challenge for the clinical microbiologist. This study was aimed to characterize the S. epidermidis isolates obtained from device related infection for their phenotypic and molecular markers of virulence and to see whether these markers can be used to differentiate the pathogenic S. epidermidis from the commensals. METHODS: Fifty five S. epidermidis isolates from various device related infections such as endophthalmitis following intra-ocular lens (IOL) implantation, intravascular (IV) catheter related sepsis and orthopaedic implant infections, were studied for slime production, biotyping, antibiotic sensitivity; and mec A and ica positivity by the recommended procedures. RESULTS: Twenty three (41.8%) isolates were multi-drug resistant, 26 (65.2%) were slime producers, 30 (54.5%) were adherent, 23 (41.8%) possessed the intercellular adhesin (ica) gene, and 28 (50.9%) harboured the mec A gene. Biotypes I and III were the commonest, most members of which were multi- drug resistant. Twenty two (73.3%) of the 30 adherent bacteria were slime producers as opposed to only 4 (16%) of the 25 non-adherent bacteria (P<0.001). A vast majority i.e. 21 (91.3%) of the 23 ica positive organisms were adherent to artificial surfaces in contrast to only 9 (28.1%) of the 32 non-ica positive organisms (P<0.001). Twenty (86.9%) of the 23 ica positive bacteria were slime producers, as opposed to only 6 (18.7%) of the 32 ica negative bacteria (P<0.001). Of the 23 multi-drug resistant isolates, 19 (82.6%) carried the mec A gene. INTERPRETATION & CONCLUSIONS: The present findings showed that ica AB and mec A were the two important virulence markers of S. epidermidis in implant infections and slime was responsible for the sessile mode of attachment on the devices.

Global alteration in gene expression profiles of deciduas from women with idiopathic recurrent pregnancy loss.

Recurrent pregnancy loss (RPL) occurs in ∼5% of women. However, the etiology is still poorly understood. Defects in decidualization of the endometrium during early pregnancy contribute to several pregnancy complications, such as pre-eclampsia and intrauterine growth restriction (IUGR), and are believed to be important in the pathogenesis of idiopathic RPL. We performed microarray analysis to identify gene expression alterations in the deciduas of idiopathic RPL patients. Control patients had one antecedent term delivery, but were undergoing dilation and curettage for current aneuploid miscarriage. Gene expression differences were evaluated using both pathway and gene ontology (GO) analysis. Selected genes were validated using quantitative reverse transcription-polymerase chain reaction (qRT-PCR). A total of 155 genes were found to be significantly dysregulated in the deciduas of RPL patients (>2-fold change, P < 0.05), with 22 genes up-regulated and 133 genes down-regulated. GO analysis linked a large percentage of genes to discrete biological functions, including immune response (23%), cell signaling (18%) and cell invasion (17.1%), and pathway analysis revealed consistent changes in both the interleukin 1 (IL-1) and IL-8 pathways. All genes in the IL-8 pathway were up-regulated while genes in the IL-1 pathway were down-regulated. Although both pathways can promote inflammation, IL-1 pathway activity is important for normal implantation. Additionally, genes known to be critical for degradation of the extracellular matrix, including matrix metalloproteinase 26 and serine peptidase inhibitor Kazal-type 1, were also highly up-regulated. In this first microarray approach to decidual gene expression in RPL patients, our data suggest that dysregulation of genes associated with cell invasion and immunity may contribute significantly to idiopathic recurrent miscarriage.

Fundus autofluorescence imaging in an ocular screening program.

Purpose. To describe integration of fundus autofluorescence (FAF) imaging into an ocular screening program. Methods. Fifty consecutive screening participants were included in this prospective pilot imaging study. Color and FAF (530/640 nm exciter/barrier filters) images were obtained with a 15.1MP Canon nonmydriatic hybrid camera. A clinician evaluated the images on site to determine need for referral. Visual acuity (VA), intraocular pressure (IOP), and ocular pathology detected by color fundus and FAF imaging modalities were recorded. Results. Mean ± SD age was 47.4 ± 17.3 years. Fifty-two percent were female and 58% African American. Twenty-seven percent had a comprehensive ocular examination within the past year. Mean VA was 20/39 in the right eye and 20/40 in the left eye. Mean IOP was 15 mmHg bilaterally. Positive color and/or FAF findings were identified in nine (18%) individuals with diabetic retinopathy or macular edema (n = 4), focal RPE defects (n = 2), age-related macular degeneration (n = 1), central serous retinopathy (n = 1), and ocular trauma (n = 1). Conclusions. FAF was successfully integrated in our ocular screening program and aided in the identification of ocular pathology. Larger studies examining the utility of this technology in screening programs may be warranted.

Slime production is essential for the adherence of Staphylococcus epidermidis in implant-related infections.

A total of 32 Staphylococcus epidermidis isolates from indwelling device-related infections such as endophthalmitis following intraocular lens (IOL) implantation, intravenous catheter-related sepsis and orthopaedic implant infections, were studied for slime production and adherence to artificial surfaces. Of these, 21 (65.6%) isolates were slime positive by the Congo Red agar method and 24 (75%) were adherent to artificial surfaces by the quantitative slime test. The majority (19 out of 24; 79.1%) of the adherent bacteria were slime producers. Antibody to slime raised in rabbits was able to inhibit the adherence of all 24 bacteria designated as adherent by our quantitative test. It seems that slime is indispensable for the sessile mode of attachment, leading further to the development of biofilms on the indwelling devices.

Secondhand smoke in public places: can Bangalore metropolitan transport corporation be a role model for effective implementation of Cigarette and Other Tobacco Products Act, 2003?

BACKGROUND: The Indian government enacted 'The cigarettes and other tobacco products act, 2003' (COTPA), which prohibits smoking in public places. AIM: To validate the efficacy of the Act of 2003, enacted by the Government of India, to prevent secondhand smoking in public places. SETTINGS AND DESIGN: The study is based on a non-random sample survey of 2,600 bus passengers carried out in the premises of three mega public road transport organizations in Karnataka state, India, in June 2007. METHODS AND MATERIAL: The information was gathered through administration of structured schedules. A sample of 1,000 each for the terminus of Bangalore Metropolitan Transport Corporation (BMTC) and Karnataka State Road Transport Corporation (KSRTC) in Bangalore and, 600 for North West Karnataka Road Transport Corporation (NWKRTC) in Hubli-Dharwad city was distributed proportionately according to the number of platforms in each terminus. STATISTICAL ANALYSIS USED: Simple Averages. RESULTS: There is some reduction in smoking in general as perceived by 69% of the passengers as compared to the scenario a year before the enactment of COTPA. The observed smoking is lower in the bus premises of BMTC where there is strict regulation, and higher in the bus premises of NWKRTC, which has not taken any regulatory measures. CONCLUSIONS: Knowing smoking is banned in public places can itself create awareness depending on the coverage extended by media and implementing an agency to reach the public. The implementation of an act depends on the willingness of stakeholders to act upon it. The implementation of COTPA as done by BMTC could well become a role model for replication elsewhere, if BMTC can strive harder to accomplish a 100% smoke-free zone.

Correlation of proteinase production with amphotericin B resistance in fungi from mycotic keratitis.

Two hundred fungal isolates (Aspergillus and Fusarium species) from mycotic keratitis were tested for in vitro susceptibilities to amphotericin B and proteinase production. Geometric mean MICs for all fungal species increased fourfold with thousandfold increase in the inoculum. The MIC(50) and MIC(90) values ranged between 3.12-6.25 and 3.12-12.5 microg/ml, respectively. Proteinase production was noted in 113 (56.5%) isolates. Ninety-eight (49%) showed MICs of > or =1.56 microg/ml that was above the criteria of > or =1 microg/ml for amphotericin B resistance (CLSI). Seventy-three (74.5%) of these 98 isolates were proteinase producers, whereas only 40 (39.2%) of the remaining 102 with low MICs (<1.56 microg/ml) were proteinase producers (p < 0.001). Proteinase seems to be an important virulence marker of filamentous fungi in mycotic keratitis, correlating significantly with amphotericin B resistance.

Effect of heme oxygenase-1 deficiency on placental development.

Heme oxygenase (HO) is the rate-limiting enzyme in the heme catabolic pathway and highly expressed in the placenta. Deficiencies in HO-1, the inducible isoform, have been associated with pregnancy disorders, such as recurrent miscarriages, intrauterine growth retardation, and pre-eclampsia. The aim of this study was to identify if a deficiency in HO-1 affects placental development using a mouse model. When HO-1 heterozygote (Het, HO-1(+/-)) mice were cross-bred, an extremely low birth rate in homozygote (Mut, HO-1(-/-)) offspring (2.4%) and small litter sizes were observed. Placentas and fetuses from Het cross-breedings were relatively smaller and weighed less than those from wild-type (WT) cross-breedings at E12.5 and E15.5. Furthermore, Het placentas had significantly less HO-1 mRNA and protein levels than WT placentas, but no significant differences in placental HO activity. Interestingly, HO-2, the constituitive HO isoform, as well as iNOS and eNOS expression were significantly upregulated in Het placentas. Histological examination showed that the junctional zone (JZ) of Het placentas were markedly thinner than those of WT placentas and appeared to be due to an increase in apoptosis. Immunohistochemistry revealed that HO-1-expressing cells were located primarily in the JZ of Het placentas, specifically in the spongiotrophoblast layer. In addition, diastolic blood pressures and plasma soluble VEGFR-1 (sFlt-1) levels were significantly elevated in pregnant Het mice. We conclude that a partial deficiency in HO-1 is associated with morphological changes in the placenta and elevations in maternal diastolic blood pressure and plasma sFlt-1 levels, despite a compensatory increase in HO-2 expression.

Fungal infections of the eye--laboratory diagnosis and treatment.

Infections of the eye give rise to severe ocular morbidity and blindness include keratitis, orbital cellulites, endophthalmitis and dacryocystitis. Corneal blindness, in developing countries is predominantly associated with infections. In India, nearly 30-35% of all culture positive infectious keratitis are caused fungi. Laboratory diagnosis mainly depends upon proper collection and transport of clinical specimens. In fungal keratitis, corneal scraping is the ideal sample, but occasionally corneal biopsy or anterior chamber aspirate may also be needed. Corneal scraping is usually by Kimura spatula, under a slit lamp examination, after anaesthetizing the cornea with topical anaesthetic like 0.4% proparcaine. Corneal biopsy is done by a minor trephining and AC aspirate using a sterile tuberculin syringe. In case of endophthalmitis, 150-200 ìl of aqueous humour is collected. Vitreous fluid (500-1000 ìl), however, is collected by pars plana vitrectomy onto sterile tuberculin syringe, the needle is then fixed to a sterile rubber bung after expelling air from the syringe. The collected sample is immediately transported to the laboratory. Swabs from the regurgitating lacrimnal sacs and wound aspirate/swabs are the ideal specimens for dacryocystitis and orbital cellulites, respectively. These samples are cultured onto SDA slants following standard procedures. The main draw back of culture is its long incubation time (5 to 14 days), though it is indispensable from the view point of the specificity. Direct examination (KOH wet mount, Gram's, Giemsa or calcofluor fluorescent staining methods) of the specimen, however, is quick and immensely helpful for ophthalmologist. The newer rapid methods, such as molecular techniques are also available and the management of patients can be according to the results obtained. With the advent of novel antifungal agents such as newer azoles and cell wall acting antifungals like echinocandins, the clinician has the wider option of selecting the therapeutic modality. In the event of the increasing reports of in vitro drug resistance to much frequently used azoles, polyenes and 5-fluorocytosines, clinical applicability of the newer antifungal agents seems to be quite promising.