Activated T cell subsets in human type 1 diabetes: evidence for expansion of the DR+ CD30+ subpopulation in new-onset disease

An important limitation in T cell studies of human autoimmune (type 1) diabetes is lack of direct access to cells infiltrating the pancreas. We hypothesized that cells recently released from the pancreas into the blood might express a characteristic combination of markers of activation. We therefore examined the recently activated circulating T cell population [CD3+, human leucocyte antigen D-related (HLA-DR+)] using cytokine production and 10 additional subset markers [CD69, CD25, CD122, CD30, CD44v6, CD57, CD71, CCR3 (CD193), CCR5 (CD195) or CXCR3 (CD183)], comparing newly diagnosed adult (ND) (age 18-40 years) patients (n=19) to patients with diabetes for >10 years [long-standing (LS), n=19] and HLA-matched controls (C, n=16). CD3+ DR+ cells were enriched by two-step immunomagnetic separation. No differences in basal or stimulated production of interleukin (IL)-4, IL-10, IL-13 or interferon (IFN)-gamma by CD3+ DR+ enriched cells were observed between the different groups of subjects. However, among the CD3+ DR+ population, significant expansions appeared to be present in the very small CD30+, CD69+ and CD122+ subpopulations. A confirmatory study was then performed using new subjects (ND=26, LS=15), three-colour flow cytometry, unseparated cells and three additional subset markers (CD38, CD134, CD4/CD25). This confirmed the expansion of the CD3+ DR+ CD30+ subpopulation in ND subjects. We conclude that a relative expansion in the T cell subpopulation with the activated phenotype CD3+ DR+ CD30+ is seen in the peripheral blood of subjects with newly diagnosed type 1 diabetes. This subpopulation represents less than 0 x 7% of circulating T cells and may provide a rich source of disease-specific T cells that can be isolated from blood.     

Assessment of retinal manifestations of Parkinson’s disease using spectral domain optical coherence tomography: A study in Indian eyes

Purpose:To assess the retinal manifestations of Parkinson’s disease using optical coherence tomography.Methods:A prospective case-control study comparing 30 eyes from 15 patients with Parkinson’s disease and 22 eyes from 11 healthy age-matched controls. Total macular subfield thickness and the thickness of the ganglion cell layer, nerve fiber layer, and peripapillary retinal nerve fiber layer were measured with spectral-domain optical coherence tomography (SD-OCT).Results:The mean age of PD patients was 68.4 years ± 10.64 (range: 46–82) and in the control group was 66.36 ± 5.22 (range: 64–68). The average disease duration in patients with PD was 6.7 ± 2.8 years (range: 2–10 years). The mean best-corrected visual acuity in PD was 20/26 and 20/20 in controls, with P = 0.0059, which was significant. Significant difference was also found in the contrast sensitivity between both groups. Structural differences in the central macular thickness (P = 0.0001), subfield thicknesses in the superior (P = 0.003), inferior (P = 0.001), nasal (P = 0.004), and temporal subfields (P = 0.017) was seen. Severe thinning of the ganglion cell layer was seen in PD patients (P = 0.000) as well as of the nerve fiber layer (P = 0.004). Peripapillary retinal nerve fiber thickness measured showed significant thinning in superotemporal (P = 0.000), superonasal (P = 0.04), inferonasal (P = 0.000), inferotemporal (P = 0.000), nasal (P = 0.000), and temporal quadrants (P = 0.000).Conclusion:Visual dysfunction was observed in patients with PD along with structural alterations on OCT, which included macular volumes, ganglion cell layer, and peripapillary retinal nerve fiber layer.     

Determination of Cyanotoxins and Prymnesins in Water,Fish Tissue,and Other Matrices: A Review

Harmful algal blooms (HABs) and their toxins are a significant and continuing threat to aquatic life in freshwater, estuarine, and coastal water ecosystems. Scientific understanding of the impacts of HABs on aquatic ecosystems has been hampered, in part, by limitations in the methodologies to measure cyanotoxins in complex matrices. This literature review discusses the methodologies currently used to measure the most commonly found freshwater cyanotoxins and prymnesins in various matrices and to assess their advantages and limitations. Identifying and quantifying cyanotoxins in surface waters, fish tissue, organs, and other matrices are crucial for risk assessment and for ensuring quality of food and water for consumption and recreational uses. This paper also summarizes currently available tissue extraction, preparation, and detection methods mentioned in previous studies that have quantified toxins in complex matrices. The structural diversity and complexity of many cyanobacterial and algal metabolites further impede accurate quantitation and structural confirmation for various cyanotoxins. Liquid chromatography–triple quadrupole mass spectrometer (LC–MS/MS) to enhance the sensitivity and selectivity of toxin analysis has become an essential tool for cyanotoxin detection and can potentially be used for the concurrent analysis of multiple toxins.     

Severe retinopathy of prematurity in big babies in India: History repeating itself?

Objective  

To describe the characteristics of babies with severe retinopathy of prematurity (ROP) attending a tertiary referral eye hospital in South India.     

Diabetic retinopathy among self reported diabetics in southern India: a population based assessment

AIMS: To estimate the prevalence of diabetic retinopathy among self reported diabetics in a population of southern India. METHODS: A cross sectional sample of subjects aged 50 years and older was selected using a cluster sampling technique from Palakkad district of Kerala state. Eligible subjects were identified through a door to door survey. Ocular examinations including visual acuity and anterior and posterior segment examinations were performed at preselected sites within clusters. History of diabetes was elicited, and height, weight, and blood pressure were measured for all subjects. RESULTS: Among the 5212 examined people (92% response rate), 68 (26.2%) of 260 people with self reported history of diabetes had diabetic retinopathy. The age-sex adjusted prevalence of diabetes among people aged 50 years and older was 5.1% (95% CI 3.9, 6.3, deff 4.33) and of diabetic retinopathy among the diabetics was 26.8% (95% CI: 19.2, 34.4, deff 1.99). Non-proliferative diabetic retinopathy (94.1%) was the most common form of retinopathy seen. Two eyes were blind (presenting vision <6/60) as a result of retinopathy. CONCLUSION: Preventive strategies have to be evolved to ensure that blindness due to diabetic retinopathy does not become a public health problem in India. Further studies are required to understand the risk factors for retinopathy and vision loss in this population.     

Optic nerve entrance coloboma associated with situs inversus

A rare case of optic nerve entrance coloboma associated with situs inversus is reported.     

Adenovirus-mediated p53 gene therapy in pediatric soft-tissue sarcoma cell lines: sensitization to cisplatin and doxorubicin

Sarcomas, or tumors of connective tissue, represent roughly 20% of childhood cancers. Although the cure rate for sarcomas in general has significantly improved in the last 10 years, there continue to be subgroups that are difficult to treat. High-grade or metastatic soft-tissue sarcomas and rhabdomyosarcomas (RMS) of the extremities remain therapeutic challenges and their prognosis is often poor. The future of sarcoma therapy will likely include molecular approaches including gene/protein expression profiling and gene-based therapy. Most sarcomas harbor defects in the p53 or pRb pathways. The tumor suppressor p53 is central to regulation of cell growth and tumor suppression and restoring wild-type p53 function in pediatric sarcomas may be of therapeutic benefit. Studies with adenoviral-mediated p53 gene transfer have been conducted in many cancer types including cervical, ovarian, prostatic and head and neck tumors. Studies of this approach, however, remain limited in pediatric cancers, including sarcomas. Using three viral constructs containing cDNA for wild-type p53, mutant p53 (C135S) and lacZ, we studied the effect of adenoviral-mediated gene therapy in four pediatric sarcoma cell lines, RD and Rh4 (RMS), Rh1 (Ewing's sarcoma) and A204 (undifferentiated sarcoma). Using the MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay, we have shown a dose-dependent decrease in cell viability 72 h post-treatment that occurs with Ad-wtp53 but not with Ad-mutp53. Cells treated with Ad-wtp53 show upregulation of the p53 downstream targets, p21(CIP1/WAF1) and bax. Growth curves demonstrate suppression of cell growth over a period of 4 days and cells treated with Ad-wtp53 demonstrate a significant increase in sensitivity to the chemotherapeutic agents, cisplatin and doxorubicin. Our results indicate that restoration of wild-type p53 function in pediatric sarcoma cells could provide a basis for novel approaches to treatment of this disease.