HPLC-LED-Induced Fluorescence Analysis of Tear Fluids: An Objective Method for Primary Open Angle Glaucoma Diagnosis.

PURPOSE: The study showcased the application of the lab-assembled HPLC-LED-IF system to analyze proteins in tear fluid samples collected from individuals diagnosed with primary open-angle glaucoma (POAG). METHODS: Clinical application of the said technique was evaluated by recording chromatograms of tear fluid samples from control and POAG subjects and by analyzing the protein profile using multivariate analysis. The data analysis methods involved are principal component analysis (PCA), Match/No-Match, and artificial neural network (ANN) based binary classification for disease diagnosis. RESULTS: Mahalanobis distance and spectral residual values calculated using a standard calibration set of clinically confirmed POAG samples for the Match/No-Match test gave 86.9% sensitivity and 81.8% specificity. ANN with leaving one out procedure has given 87.1% sensitivity and 81.8% specificity. CONCLUSION: The results of the study revealed that the utilization of a 278 nm LED excitation in the HPLC system offers good sensitivity for detecting proteins at low concentrations allowing to obtain reliable protein profiles for the diagnosis of POAG.

Highly efficient, perfect, large angular and ultrawideband solar energy absorber for UV to MIR range.

Although different materials and designs have been tried in search of the ideal as well as ultra-wideband light absorber, achieving ultra-broadband and robust unpolarized light absorption over a wide angular range has proven to be a major issue. Light-field regulation capabilities provided by optical metamaterials are a potential new technique for perfect absorbers. It is our goal to design and demonstrate an ultra-wideband solar absorber for the ultraviolet to a mid-infrared region that has an absorptivity of TE/TM light of 96.2% on average. In the visible, NIR, and MIR bands of the solar spectrum, the absorbed energy is determined to be over 97.9%, above 96.1%, and over 95%, respectively under solar radiation according to the Air Mass Index 1.5 (AM1.5) spectrum investigation. In order to achieve this wideband absorption, the TiN material ground layer is followed by the SiO2 layer, and on top of that, a Cr layer with patterned Ti-based resonators of circular and rectangular multiple patterns. More applications in integrated optoelectronic devices could benefit from the ideal solar absorber's strong absorption, large angular responses, and scalable construction.

Human tear fluid analysis for clinical applications: progress and prospects.

Introduction: Human blood and saliva are increasingly under investigation for the detection of biomarkers for early diagnosis of non-communicable (e.g.cancers) and communicable diseases like COVID-19. Exploring the potential application of human tears, an easily accessible body fluid, for the diagnosis of various diseases is the need of the hour.Areas covered: This review deals with a comprehensive account of applications of tear analysis using different techniques, their comparison and overall progress achieved till now. The techniques used for tear fluid analysis are HPLC/UPLC/SDS-PAGE, CE, etc., together with ELISA, Mass Spectrometry, etc. But, with advances in instrumentation and data processing methods, it has become easy to couple the various separation methods with highly sensitive optical techniques for the analysis of body fluids.Expert opinion: Tear analysis can provide valuable information about the health condition of the eyes since it contains several molecular constituents, and their relative concentrations may alter under abnormal conditions. Tear analysis has the advantage that it is totally non-invasive. This study recommends tear fluid as a reliable clinical sample to be probed by highly sensitive optical techniques to diagnose different health conditions, with special emphasis on eye diseases.

A comparative study of the various patterns of oro-cutaneous fungi and their sensitivity to anti fungals between HIV patients and normal healthy individuals.

BACKGROUND: Dermatological diseases are the first recognized clinical manifestation Human Immunodeficiency Virus (HIV) and Acquired Immune Deficiency Syndrome (AIDS).1, 2 The present study was undertaken to find out the clinical spectrum of the superficial mycoses, the etiological organisms and their drug sensitivity patterns among HIV positive patients and non HIV individuals attending the tertiary care hospital. METHODS: The study population was 100 HIV patients and control patients were consecutive 100 HIV negative patients. Skin scrapings and swabs were obtained from the upper back, web spaces of toes, inguinal region, dorsum of tongue. All the samples were subjected to potassium hydroxide mount and stained with Calcoflour White and were cultured. The fungi were identified on the basis of colony and microscopic features in conjunction with results of physiologic evaluation by standard phenotypic identification criteria. RESULTS: The total number of seropositive patients who had atleast one fungal infection was 57 and the total number of seronegative patients who had atleast one fungal infection was 21. In our study, fungal colonization was seen in 3.6% in clinically normal sites in retropositive patients and 1.6% in retronegatives. 76.59% in retropositive and 85.71% in retronegative patients the fungi cultured were sensitive to fluconazole. CONCLUSION: The findings of this study suggest that the skin of HIV patients may more frequently harbour common fungi even in the absence of visible clinical signs. Antifungal-resistant fungi should be kept in mind while treating fungal infections.

Anesthesia in a child with Kinsbourne syndrome: Does anesthesia technique matters?

Kinsbourne syndrome is a rare neurological paraneoplastic syndrome associated with neuroblastic tumors. There are very few literatures on its anesthetic management and interaction with anesthetic agents. The epileptogenic potential of certain anesthetic agents such as ketamine, etomidate, and meperidine might trigger opsoclonus and myoclonus and have an impact on the long-term neurological outcome. The objective of this case report is to discuss the safety of anesthetic agents and their relationship in a patient with Kinsbourne syndrome. We discuss our experience in the anesthetic management of a child with Kinsbourne syndrome with ganglioneuroblastoma in the thoracic paravertebral space.

Rare case of subcutaneous mycosis with intrathoracic extension due to Chaetomium strumarium.

A 47-year-old man presented with a 10-year history of multiple lumps over his left upper arm and shoulder and the adjoining left side of his chest and upper back. His medical history included diabetes mellitus type 2. The patient was a farmer and used to lift sacks of grains and fertilizers onto his shoulders as part of his work, although he did not recollect any history of specific trauma. Skin biopsy revealed granulomatous reaction with Splendore-Hoeppli phenomenon, while periodic-acid-Schiff and Grocott-Gomori stains confirmed fungal elements. Sabouraud agar grew Chaetomium species, and lactophenol blue mount confirmed the fungus as Chaetomium strumarium. Radiography and computed tomography of the chest revealed intrathoracic extension of the mycetoma. The patient responded well to treatment with oral Itraconazole. Subcutaneous mycosis due to C. strumarium is rarely reported in the literature, and the intrathoracic extension makes it an even rarer entity.

Immune reconstitution inflammatory syndrome in leprosy.

Immune reconstitution inflammatory syndrome (IRIS) is a paradoxical deterioration in the clinical status of a patient infected with human immunodeficiency virus (HIV) on highly active antiretroviral treatment (HAART). The immune suppression caused by the virus can initially suppress the clinical manifestations of leprosy which can then be unmasked after treatment with HAART or an inflammatory reaction can occur in the initial months of therapy, resulting from dysregulated recovery of immunity to specific antigens. Both these conditions are identified as IRIS in leprosy. Though this syndrome is a widely recognized entity presently, there is still a lack of universally acceptable diagnostic criteria for the condition. The first case published case of leprosy- associated immune reconstitution disease was reported in 2003 and about 47 confirmed cases of IRIS in leprosy have been reported since then, mostly from Brazil and India. Anti-inflammatory drugs and steroids are the drugs of choice in inflammatory episodes with continuation of antiretroviral therapy. With increasing affordability of antiretroviral therapy, clinicians will put more and more number of human immunodeficiency virus infected patients on therapy and hence an increase in the incidence of IRIS is expected. Therefore, it is important to understand all facets of this syndrome which is becoming more prevalent with each passing day.

Evaluation of DNA damage induction and repair inhibition in welders exposed to hexavalent chromium.

The soluble hexavalent chromium Cr (VI) used in industrial welding is an environmental contaminant widely recognized to act as a carcinogen, mutagen and teratogen towards humans and animals. The carcinogenic potential of metals is a major issue in defining human health risk from exposure. In the present investigation, 93 welders and 60 control subjects with similar mean ages, smoking prevalences and alcohol consumption were enrolled for DNA damage analysis in blood leucocytes by Micronucleus assay (MN) and the Comet assay. DNA repair inhibition was also analyzed by assessing XPD gene polymorphism. Welders showed a significant increase in micronucleated cells compared to controls with respect to their smoking habits and alcohol consumption, age and years of exposure (P<0.05). Results indicated that the welders had a larger mean comet tail length than that of the controls (P<0.05). The current study suggested that chronic occupational exposure to Cr (VI) during welding could lead to increased levels of DNA damage and repair inhibition.

Irritant Dermatitis to Staphylinid Beetle in Indian Troops in Congo.

BACKGROUND: Contact dermatitis to Staphylinid beetle is a well known entity worldwide. Paederus sabaeus (Econda) species of this insect was responsible for a widespread occurrence of this dermatitis in United Nations (UN) troops posted in Congo. This study was undertaken to observe the various aspects of this unique dermatitis in the mission area. METHODS: All clinically diagnosed cases of contact dermatitis to Econda occurring in Indian troops posted to UN mission in Congo during the study period were included. Their epidemiological and clinical characteristics with treatment options were studied in detail. RESULT: A wide range of dermatological manifestations were seen in the subjects under study. Few of the manifestations observed in the study have not been reported earlier in literature. Most of the reactions occurred on the exposed areas. Treatment options were guided by the site and the severity of the reaction. CONCLUSION: Contact dermatitis to Staphylinid beetle can mimic various other dermatoses and has to be considered in the differential diagnosis in all dermatological consultations during the peak season in the Congo mission area. The insect was found to have certain definite behavioural patterns, the knowledge of which would help in preventing this dermatosis.

Serum protein profile study of normal and cervical cancer subjects by high performance liquid chromatography with laser-induced fluorescence.

High performance liquid chromatography with high sensitivity laser-induced fluorescence detection is used to study the protein profiles of serum samples from healthy volunteers and cervical cancer subjects. The protein profiles are subjected to principal component analysis (PCA). PCA shows that the large number of chromatograms of a given class of serum samples--say normal/malignant--can be expressed in terms of a small number of factors (principal components). Three parameters--scores of the factors, squared residuals, and Mahalanobis distance--are derived from PCA. The parameters are observed to have a narrow range for protein profiles of standard calibration sets formed from groups of clinically confirmed normal/malignant classes. Limit tests using match/no match of the parameters of any test sample with parameters derived for the standard calibration sets give very good discrimination between malignant and normal samples with high sensitivity (approximately 100%) aand specificity (approximately 94%).

Omega-3 fatty acid effect on alveolar bone loss in rats.

Gingival inflammation and alveolar bone resorption are hallmarks of adult periodontitis, elicited in response to oral micro-organisms such as Porphyromonas gingivalis. We hypothesized that omega (omega)-3 fatty acids (FA) dietary supplementation would modulate inflammatory reactions leading to periodontal disease in infected rats. Rats were fed fish oil (omega-3 FA) or corn oil (n-6 FA) diets for 22 weeks and were infected with P. gingivalis. Rats on the omega-3 FA diet exhibited elevated serum levels of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), documenting diet-induced changes. PCR analyses demonstrated that rats were orally colonized by P. gingivalis; increased IgG antibody levels substantiated this infection. P. gingivalis-infected rats treated with omega-3 FA had significantly less alveolar bone resorption. These results demonstrated the effectiveness of an omega-3 FA-supplemented diet in modulating alveolar bone resorption following P. gingivalis infection, and supported that omega-3 FA may be a useful adjunct in the treatment of periodontal disease.

Combined Fourier transform infrared and Raman spectroscopic approach for identification of multidrug resistance phenotype in cancer cell lines.

Cancer cells escape cytotoxic effects of anticancer drugs by a process known as multidrug resistance (MDR). Identification of cell status by less time-consuming methods can be extremely useful in patient management and treatment. This study aims at evaluating the potentials of vibrational spectroscopic methods to perform cell typing and to differentiate between sensitive and resistant human cancer cell lines, in particular those that exhibit the MDR phenotype. Micro-Raman and Fourier transform infrared (FTIR) spectra have been acquired from the sensitive promyelocytic HL60 leukemia cell line and two of its subclones resistant to doxorubicin (HL60/DOX) and daunorubicin (HL60/DNR), and from the sensitive MCF7 breast cancer cell line and its MDR counterpart resistant to verapamil (MCF7/VP). Principal components analysis (PCA) was employed for spectral comparison and classification. Our data show that cell typing was feasible with both methods, giving two distinct clusters for HL60- and MCF7-sensitive cells. In addition, phenotyping of HL60 cells, i.e., discriminating between the sensitive and MDR phenotypes, was attempted by both methods. FTIR could not only delineate between the sensitive and resistant HL60 cells, but also gave two distinct clusters for the resistant cells, which required a two-step procedure with Raman spectra. In the case of MCF7 cell lines, both the sensitive and resistant phenotypes could be differentiated very efficiently by PCA analysis of their FTIR and Raman point spectra. These results indicate the prospective applicability of FTIR and micro-Raman approaches in the differentiation of cell types as well as characterization of the cell status, such as the MDR phenotype exhibited in resistant leukemia cell lines like HL60 and MCF7.

Optical diagnosis of cervical cancer by fluorescence spectroscopy technique.

In the present work, we examine normal and malignant stage IIIB cervical tissue by laser induced fluorescence, with 2 different objectives. (i) Development of the fluorescence spectroscopy technique as a standard optical method for discrimination of normal and malignant tissue samples and, (ii) Optimization of the technique by the method of matching of a sample spectrum with calibration sets of spectra of pathologically certified samples. Laser-induced fluorescence spectra were measured using samples from 62 subjects at different excitation wavelengths. Principal component analysis (PCA) of spectra and intensity ratios of curve-resolved fluorescence peaks were tested for discrimination. It was found that PCA of total fluorescence at 325 nm excitation gives specificity and sensitivity over 95%. Use of calibration sets of spectra of histo-pathologically certified samples combined with PCA for matching and pass/fail classification of test samples is shown to have high sensitivity/specificity for routine diagnostic purposes as well as for possible staging of the disease. Further, the multi-component origin of the fluorescence spectra is illustrated by curve resolution and fluorescence spectra of separated proteins of tissue homogenates.

Cervical intraepithelial neoplasia and human papilloma virus infection in renal transplant recipients.

An association between human papilloma virus (HPV) infection and cervical intraepithelial neoplasia has been well established Renal transplant recipients on long term immunosuppression are prone to viral infection. It is possible that there may be an increased prevalence of papilloma virus infection and associated cervical intraepithelial neoplasia in these women. Prospective study of 42 renal transplant recipients and 41 age and parity matched controls was undertaken to determine whether HPV infection and cervical intraepithelial neoplasia (CIN) occurred more often in renal allograft recipients and to assess the relative risk. All women underwent Pap smear, colposcopy, histological examination of biopsy specimen and polymerase chain reaction for HPV 16. Cytology did not pick up HPV infection in any of the women. Colposcopy revealed HPV infection in 15 and CIN in 14 women in the immunosuppressed group and HPV in 7 and CIN in 5 women in control group. Histological evidence of HPV was found in 24 and CIN in 10 women in immunosuppressed group and HPV infection in 13 and CIN in 3 women in control group, giving an odds ratio of 6.1. More women in the immunosuppressed group had CIN of higher degree as well. PCR revealed infection by HPV 16 in 17 cases and 14 controls giving an odds ratio of 1.3. Therefore renal allograft recipients on immunosuppression should be screened by colposcopy and directed biopsy at regular intervals.

Brucellosis--cause of abortion in sheep and its public health significance.

Present study was undertaken to determine the association of brucellosis with abortions occurring naturally in sheep at an organized local sheep breeding farm. A total of 15 strains of Brucella melitensis biovar I were isolated from the abortion material. Serologically the aborted ewes were positive for brucellosis by one or more tests. During acute infection (abortion), standard tube agglutination test (SAT) detected more positive reactors (70.7%) while counter immunoelectrophoresis (CIE) detected more positive reactors (33.9%) in chronic infection (in-contact and apparently healthy sheep). Personnel handling the abortion material at the farm were found positive clinically as well as serologically for brucellosis. These observations suggest the zoonotic importance of brucellosis.