Preparation and uses of amniotic membrane for ocular surface reconstruction

Background: The use of human amniotic membrane transplantation is rapidly increasing for the management of various ocular surface disorders. Despite its numerous advantages, amniotic membrane is not widely available due to the lack of awareness among ophthalmologists regarding its preparation and preservation techniques. Purpose: To provide an instructional video demonstrating the technique of harvesting, preparation, preservation of human amniotic membrane, and its uses in the management of ocular surface diseases. Synopsis: The amniotic membrane is the innermost layer of the human placenta. Properties of the amniotic membrane, like the presence of various growth factors, anti?inflammatory and anti?angiogenic factors, and its low immunogenicity, contribute to its ability to promote epithelial growth and differentiation with the reduction in fibrosis during healing. To harvest amniotic membrane, informed consent is obtained from a patient undergoing an elective caesarian section, and the donor is screened to exclude the risk of infections. Under sterile precautions, the amniotic membrane is separated from the chorion and washed free of blood clots. With the epithelial surface up, the amniotic membrane is spread uniformly without folds or tears on individually sterilized 0.22 ?m nitrocellulose filter papers of the required sizes. The prepared filter paper with the adherent amniotic membrane is placed in freshly prepared Dulbecco’s Modified Eagle’s Medium and stored at ?80°C. The amniotic membrane can be used for surgical procedures like symblepharon release, pterygium or dermoid excision, perforated ulcers, nonhealing epithelial defects, etc. Highlights: Instructional videos demonstrating the preparation and preservation of amniotic membrane are very sparse. This video clearly explains how any ophthalmologist can learn to prepare and preserve the human amniotic membrane and gives a glimpse of its properties, advantages, and scope of use

Retrospective study to improve early initiation of breastfeeding in newborns

Background: Breast milk is the best nutrition to every new born baby. WHO recommends that all new-borns should be breastfed within one hour of birth and then on demand. Exclusive breast feeding i.e. avoiding any other feeds till 6 months of age, is best for the baby. Despite these guidelines, our national average of initiating breastfeeding within the first hour is less than 50% as per NFHS 2015-2016. The objective was to study the existing rate of early initiation of breast feeding in the study hospital.Methods: Retrospective data collection from 2015 to 2017 was used for this study. Mothers were educated about colostrum and benefits of early initiation of breastfeeding in their third trimester and during their baby shower.Results: Early initiation of breast feeding was successful among 2624 babies out of 2835 babies who were born in the study period.Conclusions: As compared to national averages, the study hospital had a superior percentage of babies being initiated early on breastfeeding. We need to educate and motivate the society to encourage exclusive breast feeding for all babies. Professional help in the form of dedicated lactation consultants of guidelines will help mothers to breast feed their babies sooner and for longer. Busting local myths and harmful practices is the need of the hour.

A study on efficacy of oral administration of colostrum fortified lactoferrin in the weight gain of low birth weight infants

Background: Low birth weight babies is the result of being small for gestational age (i.e. under 10th percentile of the reference population) and preterm (i.e. before 37 weeks of gestation. Preterm infants have higher protein, calorie requirement, which cannot be met with unfortified breast milk feeds.Methods: This study was conducted prospectively in the Department of Pediatrics, Rajah Muthiah Medical College and Hospital, Chidambaram from October 2016 to April 2018. 20 low birth weight babies were selected by randomization technique and they were administered with 2 grams of oral colostrum fortified lactoferrin daily for 4 weeks. The rate of weight gain, length gain and head circumference gain were assessed prospectively for a period of 4 weeks. They were analysed statistically by ANOVA and tabulated. Babies weighing less than 2.5 kg and on exclusive breast feeding, were included. Babies more than 2.5kg and less than 1kg, at risk for neonatal sepsis, congenital heart disease, necrotizing enterocolitis were excluded from the study.Results: In the present study, 20 low birth weight babies were included. 11 maternal risk factors were identified. Out of which anemia stands as a single risk factor. The rate of weight gain in oral colostrum fortified lactoferrin, is 320 grams with P value of 0.703 which is not statistically significant. The rate of length gain is 3.3cm, p value is 0.093 which is statistically insignificant. The rate of head circumference gain in oral colostrum fortified lactoferrin is 2.8cm, with the p-value of 0.001 which is statistically significant.Conclusions: The rate of weight and length gain was normal, but the rate of head circumference gain is significantly increased in oral colostrum fortified lactoferrin supplementation.

A prospective study on the effects and prevalence of maternal, fetal and placental risk factors on low birth weight neonates

Background: Maternal, foetal and placental risk factors have a causative effect in the prematurity and failure to thrive in the early infantile period. Timely diagnosis helps in the anticipation of complications specific to risk factors and managing by impairing their harmful effects on the growth and development of the baby.Methods: The study was conducted prospectively in Department of Paediatrics, Rajah Muthiah Medical College and Hospital, Chidambaram from January 2018 to June 2018. 361 Low birth weight babies were included. Neonates (n=361) weighing less than 2.5 kilograms with parental informed consent are included, whereas those with severe congenital anomaly, systemic disease or infection were excluded from the study. Various known maternal, placental and foetal risk factors are observed during the course of the study. The gestational age of the neonates was estimated using modified Dubowitz scoring. Other factors like mode of delivery of the babies were also noted. Statistical analysis (descriptive statistics) was done by using Microsoft Word 2010.Results: Out of 1040 babies born from January to July 2018, 341 babies weight less than 2.5 kg. Out of which 162 (44.88%) babies were born preterm. Of the full term births 52% were low birth weights The risk factors of the neonates are not mutually exclusive. In the present study 28 maternal risk factors were studied. The prevalence of maternal, fetal and placental risk factors was 85.32%, 74.79% and 6.65% respectively.Conclusions: Period prevalence of low birth weight babies and prematurity is 34.71% and 15.58% respectively. Risk factors are not independent to each other with maternal and foetal risk factors have almost have an effect on nearly three fourths of low birth weight neonates.

Association between human leukocyte antigen-DRB1 and human leukocyte antigen-DQB1 alleles and pemphigus vulgaris in Indian patients: A case–control study

Background: HLA-DRB1*04, -DRB1*08, -DRB1*14, -DQB1*03 and -DQB1*05 are reported to have significant association with pemphigus vulgaris; however, this is partially dependent on ethnicity. This study was done to determine the HLA-DR and -DQ types prevalent in Indian patients with pemphigus vulgaris. Methods: A prospective case–control study was done for a period of 9 months in Christian Medical College Vellore, India. HLA typing was done by PCR-SSOP method in 50 cases and 50 healthy controls. Allele frequencies in cases and controls were compared and odds ratios with 95% confidence interval were calculated. Results: The mean age of the patients (29 females, 21 males) and that of controls (36 males, 14 females) were 41.3 ± 13.65 and 35.42 ± 11.09 years, respectively. HLA-DRB1*14 was present in 47 patients and 18 controls (OR, 27.85; 95% CI, 7.57–102.42) and HLA-DQB1*05 was seen in 47 patients and 24 controls (OR, 16.97; 95% CI, 4.66–61.80). The haplotype DRB1*14, DQB1*05 was present in 44 patients and 14 controls (OR, 18.86; 95% CI, 6.58–54.05). DRB1*15 was present in 7 cases and 16 controls (OR, 0.35; 95% CI, 0.13–0.94) and DQB1*06 was present in 8 cases and 19 controls (OR, 0.31; 95% CI, 0.12–0.80). HLA-DQB1*03 was associated with significantly higher pemphigus disease area index scores. Limitations: The main limitations were that the numbers studied were small as the study was conducted at a single center, and the haplotype analysis was limited only to the proband. PDAI scores could have been influenced by prior treatment. Conclusion: There was a significant association between HLA-DRB1*14 and HLA-DQB1*05 and pemphigus vulgaris in our patients. A negative association was seen with DRB1*15 and DQB1*06.

Thymoquinone inhibits the migration of mouse neuroblastoma (Neuro-2a) cells by down-regulating MMP-2 and MMP-9 / 中国天然药物

Thymoquinone (TQ), an active component derived from the medial plant Nigella sativa, has been used for medical purposes for more than 2 000 years. Recent studies have reported that TQ blocked angiogenesis in animal model and reduced migration, adhesion, and invasion of glioblastoma cells. We have recently shown that TQ could exhibit a potent cytotoxic effect and induce apoptosis in mouse neuroblastoma (Neuro-2a) cells. In the present study, TQ treatment markedly decreased the adhesion and migration of Neuro-2a cells. TQ down-regulated MMP-2 and MMP-9 protein expression and mRNA levels and their activities. Furthermore, TQ significantly down-regulated the protein expression of transcription factor NF-κB (p65) but not significantly altered the expression of N-Myc. Taken together, our data indicated that TQ's inhibitory effect on the migration of Neuro-2a cells was mediated through the suppression of MMP-2 and MMP-9 expression, suggesting that TQ treatment can be a promising therapeutic strategy for human malignant neuroblastoma.

Effects of Antiretroviral Therapy on the Survival of Human Immunodeficiency Virus-positive Adult Patients in Andhra Pradesh, India: A Retrospective Cohort Study, 2007-2013 / 예방의학회지

OBJECTIVES: The survival outcomes of antiretroviral treatment (ART) programs have not been systematically evaluated at the state level in India. This retrospective study assessed the survival rates and factors associated with survival among adult human immunodeficiency virus (HIV)-infected patients in Andhra Pradesh, India. METHODS: The present study used data from 139 679 HIV patients aged ≥15 years on ART who were registered from 2007 to 2011 and were followed up through December 2013. The primary end point was death of the patient. Mortality densities (per 1000 person-years) were calculated. Kaplan-Meier and Cox-regression models were used to estimate survival and explore the factors associated with survival. RESULTS: The overall median follow-up time was 16.0 months (2.0 months for the deceased and 14.0 months for those lost to follow-up). Approximately 13.2% of those newly initiated on ART died during follow-up. Of those deaths, 56% occurred in the first three months. The crude mortality rate was 80.9 per 1000 person-years at risk. The CD4 count (adjusted hazard ratio [aHR],4.88; 95% confidence interval [CI], 4.36 to 5.46 for 350 cells/mm³), functional status (aHR, 3.05; 95% CI, 2.82 to 3.30 for bedridden vs. normal), and body weight (aHR, 3.69; 95% CI, 3.42 to 3.97 for 60 kg) were strongly associated with the survival of HIV patients. CONCLUSIONS: The study findings revealed that high mortality was observed within the first three months of ART initiation. Patients with poor baseline clinical characteristics had a higher risk of mortality. Expanded testing and counseling should be encouraged, with the goal of ensuring early enrollment into the program followed by the initiation of ART in HIV-infected patients.

Green synthesis of titanium dioxide nanoparticles using Psidium guajava extract and its antibacterial and antioxidant properties

OBJECTIVE@#To determine the efficacies of antibacterial and antioxidant activities of aqueous leaf extract of Psidium guajava mediated biosynthesis of titanium dioxide nanoparticles (TiO2 NPs).@*METHODS@#Synthesized TiO2 NPs were tested by disc diffusion method against against human pathogenic bacteria. The total antioxidant activity and phenolic content (Folin-Ciocalteau method) of synthesized TiO2 NPs and aqueous plant extract were determined. The scavenging radicals were estimated by DPPH method. The synthesized TiO2 NPs were characterized by XRD, FTIR, FESEM and EDX.@*RESULTS@#FTIR spectra of synthesized TiO2 NPs exhibited prominent peaks at 3 410 cm(-1) (alkynes), 1 578 cm(-1), 1 451 cm(-1) (alkanes), and 1 123 cm(-1)(C-O absorption). The morphological characterization of synthesized TiO2 NPs was analysed by FESEM which showed spherical shape and clusters with an average size of 32.58 nm. The maximum zone of inhibition was observed in the synthesized TiO2 NPs (20 μg/mL) against Staphylococcus aureus (25 mm) and Escherichia coli (23 mm). The synthesized TiO2 NPs showed more antibacterial activity than the standard antibiotic disk, tetracycline which drastically reduces the chances for the development of antibiotics resistance of bacterial species. The plant aqueous extract and synthesized TiO2 NPs were found to possess maximum antioxidant activity when compared with ascorbic acid. The content of phenolic compounds (mg/g) in leaf aqueous extract and synthesized TiO2 NPs were found to be 85.4 and 18.3 mgTA/g, respectively.@*CONCLUSIONS@#Green synthesized TiO2 NPs provides a promising approach can satisfy the requirement of large-scale industrial production bearing the advantage of low-cost, eco-friendly and reproducible.

Green synthesis of titanium dioxide nanoparticles using Psidium guajava extract and its antibacterial and antioxidant properties

Objective: To determine the efficacies of antibacterial and antioxidant activities of aqueous leaf extract of Psidium guajava mediated biosynthesis of titanium dioxide nanoparticles (TiO

Acaricidal activity of synthesized titanium dioxide nanoparticles using Calotropis gigantea against Rhipicephalus microplus and Haemaphysalis bispinosa

OBJECTIVE@#To assess the acaricidal activity of titanium dioxide nanoparticles (TiO2 NPs) synthesized from flower aqueous extract of Calotropis gigantea(C. gigantea) against the larvae of Rhipicephalus (Boophilus) microplus [R. (B.) microplus] and the adult of Haemaphysalis bispinosa (H. bispinosa).@*METHODS@#The lyophilized C. gigantea flower aqueous extract of 50 mg was added with 100 mL of TiO(OH)2 (10 mM) and magnetically stirred for 6 h. Synthesized TiO2 NPs were characterized by X-ray diffraction (XRD), Fourier transform infrared spectroscopy (FTIR), Scanning electron microscopy (SEM), and Energy dispersive X-ray spectroscopy (EDX). The synthesised TiO2 NPs were tested against the larvae of R. (B.) microplus and adult of H. bispinosa were exposed to filter paper impregnated method.@*RESULTS@#XRD confirmed the crystalline nature of the nanoparticles with the mean size of 10.52 nm. The functional groups for synthesized TiO2 NPs were 1 405.19, and 1 053.45 cm(-1) for -NH2 bending, primary amines and amides and 1 053.84 and 1 078.45 cm(-1) for C-O. SEM micrographs of the synthesized TiO2 NPs showed the aggregated and spherical in shape. The maximum efficacy was observed in the aqueous flower extract of C. gigantea and synthesized TiO2 NPs against R. (B.) microplus (LC50=24.63 and 5.43 mg/L and r(2)=0.960 and 0.988) and against H. bispinosa (LC50= 35.22 and 9.15 mg/L and r(2) = 0.969 and 0.969), respectively.@*CONCLUSIONS@#The synthesized TiO2 NPs were highly stable and had significant acaricidal activity against the larvae of R. (B.) microplus and adult of H. bispinosa. This study provides the first report of synthesized TiO2 NPs and possessed excellent anti-parasitic activity.

Anaplastic lymphoma kinase-positive primary cutaneous anaplastic large cell lymphoma- Is it a new variant.

Anaplastic large cell cutaneous lymphomas are clinically and pathologically heterogeneous, CD30 + (Ki-1) lymphoproliferative disorders. The importance of anaplastic lymphoma kinase (ALK) positivity is well known in the prognosis of primary systemic anaplastic large cell cutaneous lymphomas; however, the same in primary cutaneous anaplastic large cell cutaneous lymphomas is not much clear. Herein we report a 65-year-old male with an 18-month history of minimally pruritic localized nodulo-plaque lesion over lower back. Histology revealed cutaneous large cell lymphoma and immunohistochemical staining showed positivity for CD30, CD3 and ALK. The role of ALK positivity in pcALCL is discussed in this article.

Keratosis follicularis spinulosa decalvans in a female.

Keratosis follicularis spinulosa decalvans (KFSD), is a rare follicular syndrome associated with widespread keratosis pilaris and progressive scarring alopecia. This genodermatoses often starts at infancy or early childhood with an X-linked mode of inheritance. Males are predominantly affected and females frequently show no disease or only a mild form. We describe this not so common entity of KFSD in a nine year old female child.

Paraneoplastic multicentric reticulohistiocytosis: A clinicopathologic challenge.

Multicentric Reticulohistiocytosis (MRH) is a rare, systemic non-Langerhans cell histiocytosis (non-LCH) with prominent joint and skin manifestations. It is mostly self limiting. However, 15-30% of the cases are associated with malignancy and carry a poor prognosis. We report the case of a 42-year-old man who presented with multiple reddish-brown papules that on biopsy showed aggregates of oncocytic histiocytes with several multinucleate giant cells. Immunostains were positive for CD 68, CD 45 and were negative for S-100, CD1a. An impression of multicentric reticulohistiocytosis (MRH) was made, with the recommendation to screen for malignancy. Electron microscopy of the skin lesions showed features consistent with non-Langerhans cell histiocytosis. The patient was later diagnosed with acute myeloid leukemia at a follow-up visit several months later. Thus, it appears prudent to screen and follow-up adults with MRH, to identify an underlying malignant condition.

Berardinelli-Seip syndrome in a 6-year-old boy.

A 6-year-old boy presented with abnormal habitus since birth, delayed language development, history of frequent falls since 9 months, and fever since 1 week. He was found to have hyperandrogenic features, generalized paucity of fat, generalized muscular overdevelopment, and brownish pigmentation over the flexural creases. Skin biopsy demonstrated features suggestive of acanthosis nigricans with an absence of subcutaneous tissue. After further investigation, a diagnosis of Berardinelli-Seip syndrome with bilateral pneumonia and generalized tonic clonic seizures was made. Clinical features, histopathology, differential diagnosis, and prognosis of this rare disorder have been discussed.

Familial dyskeratotic comedones.

Familial dyskeratotic comedones (FDC) is a rare autosomal dominant inherited condition, characterized by widespread, symmetrically scattered, comedone-like, hyperkeratotic papules, which are cosmetically unappealing. These lesions appear around puberty and show a predilection to involve the trunk, arms and face. The lesions are asymptomatic and gradually worsen with time. Histology shows invagination of the epidermis with a lamellar keratinous plug and focal evidence of dyskeratosis. This condition is generally refractory to therapy. We report here two cases with this rare disorder who had a strong familial history of the same disorder.

Erythromelanosis follicularis faciei et colli: relationship with keratosis pilaris.

Erythromelanosis follicularis faciei et colli (EFF) is an unusual condition characterized by the triad of hyperpigmentation, follicular plugging and erythema of face and neck. This is less common in women and familial case reports are few. We report EFF in three siblings in an Indian family, two of whom are females. The possibility of this condition being genetically related to keratosis pilaris as well as being a variant of keratosis rubra pilaris is also discussed.

Prolonged exposure of human embryonic stem cells to heat shock induces necrotic cell death

We investigated the effects of prolonged heat shock treatment on human embryonic stem cell (hESC) viability. The hESC viability steadily declined with longer exposure to heat shock treatment (43ºC). After 4 h of exposure to heat shock at 43ºC, only 56.2 ± 1.5% of cells were viable. Viability subsequently declined to 37.0 ± 3.3% and 3.5 ± 0.7% after 8 h and 16 h, respectively of heat shock treatment at 43ºC. Transmission electron micrographs showed that the morphology of the dead/dying cells after heat shock treatment was characteristic of cellular necrosis with an uncondensed chromatin and a non-intact plasma membrane. This was further confirmed by flow cytometry analysis which showed that the DNA of the dead/ dying cells was still mostly intact, unlike the characteristic DNA fragmentation observed with apoptotic cells. In conclusion, prolonged exposure to heat shock treatment was detrimental to hESC viability. Hence, any future protocols developed for either the heat shock pre-conditioning of hESC prior to transplantation or for the temporary expression of specific genes with heat shock-responsive promoters should take these results into account; to achieve an optimal balance between the duration of heat shock exposure and the attainment of the desired effects.

Fine needle aspiration cytology of trichofolliculoma: a case report.

The cytomorphology of skin adnexal tumours has not been described extensively in literature. We report the first case of trichofolliculoma, a hair follicle hamartoma, which was subsequently diagnosed by excision biopsy. A 19 year old woman presented with a nodule on the dorsal aspect of the finger which was clinically suspected to be a myxoid cyst. FNA showed several cohesive as well as arborising branched groups of squamous cells with evidence of keratinisation. The background had dispersed and loosely clustered sebaceous cells. The excision biopsy revealed a trichofolliculoma. Cytology is a useful tool in the diagnosis of skin adnexal tumours. The finding of cohesive, branching keratinized squamous cell clusters admixed with sebaceous cells may suggest a diagnosis of trichofolliculoma, as was seen in the present case.

Cutaneous vasculitis--a dynamic process posing diagnostic challenge.

AIM: To characterize the clinicopathologic features and to assess the therapeutic outcome in cutaneous vasculitis. MATERIAL AND METHODS: Fifty biopsy proven cases of cutaneous vasculitis seen between January 1998 and July 1999 were studied. RESULTS: The commonest presentation was palpable purpura. The site most commonly affected was the extremity, irrespective of the age (adults - 40 and children - 10) and sex. The histopathologic picture ranged from an acute to chronic process, which besides the classic picture included bullous presentation, granulomatous histology and nonspecific features. Clinical correlation and investigations including direct immunofluorescence (DIF) were required to differentiate primary from secondary vasculitis (SLE-4, dermatomyositis-2, rheumatoid artritis-1, HIV-1, septicaemia-1 and drug reaction 2). DIF was diagnostic in 13 out of 21 cases providing evidence of an immune-mediated pathogenesis. Drugs used in the treatment included dapsone, colchicine, pentoxyphiline and steroids. CONCLUSIONS: The clinical picture and outcome of primary cutaneous vasculitis were benign while the prognosis of secondary vasculitis depended on the primary disease, irrespective of the histopathological picture.