Pluripotent very small embryonic-like stem cells get discarded during cord blood and bone marrow processing

The recent letter by Ivanovic commenting on work published by our group on very small embryonic-like stem cells (VSELs) in cord blood and bone marrow raises important questions on the properties and regenerative potential of VSELs. Similar to embryonic stem cells, being pluripotent by nature, VSELs are expected to have maximum regenerative potential compared with adult stem cells with limited "plasticity." We propose that both hematopoietic (HSCs) and mesenchymal (MSCs) stem cells with cytoplasmic OCT-4 are possibly descendants "progenitors" derived from VSELs with nuclear OCT-4. Being pluripotent and quiescent by nature, VSELs may serve as a good autologous source of pluripotent stem cells (with minimal risk of teratoma formation) for regenerative medicine.     

Spuriously high prevalence of prediabetes diagnosed by HbA(1c) in young indians partly explained by hematological factors and iron deficiency anemia

OBJECTIVE

To examine the influence of glycemic and nonglycemic parameters on HbA_1c concentrations in young adults, the majority of whom had normal glucose tolerance.

RESEARCH DESIGN AND METHODS

We compared the diagnosis of normal glucose tolerance, prediabetes, and diabetes between a standard oral glucose tolerance test (OGTT; World Health Organization 2006 criteria) and HbA_1c concentrations (American Diabetes Association [ADA] 2009 criteria) in 116 young adults (average age 21.6 years) from the Pune Children’s Study. We also studied the contribution of glycemic and nonglycemic determinants to HbA_1c concentrations.

RESULTS

The OGTT showed that 7.8% of participants were prediabetic and 2.6% were diabetic. By ADA HbA_1c criteria, 23.3% were prediabetic and 2.6% were diabetic. The negative predictive value of HbA_1c was 93% and the positive predictive value was 20% (only 20% had prediabetes or diabetes according to the OGTT; this figure was 7% in anemic participants). Of participants, 34% were anemic, 37% were iron deficient (ferritin <15 ng/mL), 40% were vitamin B₁₂ deficient (<150 pmol/L), and 22% were folate deficient (<7 nmol/L). On multiple linear regression analysis, HbA_1c was predicted by higher 2-h glucose (R² = 25.6%) and lower hemoglobin (R² = 7.7%). When hematological parameters were replaced by ferritin, vitamin B₁₂, and folate, HbA_1c was predicted by higher glycemia (R² = 25.6%) and lower ferritin (R² = 4.3%).

CONCLUSIONS

The use of HbA_1c to diagnose prediabetes and diabetes in iron-deficient populations may lead to a spuriously exaggerated prevalence. Further investigation is required before using HbA_1c as a screening tool in nutritionally compromised populations.The use of HbA_1c to diagnose prediabetes and diabetes is an attractive option in prospective epidemiological studies because it may avoid the need for repeated oral glucose tolerance tests (OGTTs). The American Diabetes Association (ADA) and World Health Organization (WHO) have recently approved the use of HbA_1c for screening and diagnosis of diabetes (1–3). Both organizations have suggested that concentrations ≥6.5% be considered diabetes, and the ADA has suggested 5.7–6.4% as diagnostic of prediabetes (3).The concentration of HbA_1c depends on not only prevailing glycemia but also the life span of erythrocytes. Nutritional deficiencies are a major factor affecting erythrocyte survival. Among these, iron deficiency is the most common and affects >50% of the world’s population (4). Previous studies show that iron deficiency increases erythrocyte survival and therefore disproportionately elevates HbA_1c concentrations at a given glycemic level (5,6). These were small studies in nondiabetic subjects. There is one similar report in type 1 diabetic patients (7). WHO and ADA have acknowledged this limitation of using HbA_1c in the diagnosis of prediabetes and diabetes in nutritionally compromised populations, but not the magnitude of the effect.In the current study, we aimed to investigate the diagnostic performance of HbA_1c against a standard OGTT in young adults in a prospective birth cohort (Pune Children’s Study [PCS]) and study the influence of hematological, nutritional, and other factors on HbA_1c concentrations.     

Elevated plasma gas6 levels are associated with venous thromboembolic disease

Growth arrest-specific 6 (gas6), a novel vitamin K-dependent protein, has been demonstrated to have a role in thrombus stabilization as gas6 null mice are resistant to lethal venous and arterial thrombosis. However, the association between gas6 and venous thromboembolism has not been elucidated in humans. The present study aims to assess the role of gas6 in human venous thromboembolic (VTE) disease. Using a highly specific ELISA method, we measured plasma levels of gas6 in plasma samples obtained from 279 patients with VTE and 79 healthy volunteers. Medication history, comorbid conditions and VTE characteristics were documented. Mean gas6 levels were higher in patients with VTE as compared to healthy volunteers, being 46 ±11 ng/ml and 35 ±6.4 ng/ml respectively (P < 0.001). Odds ratios (OR) for VTE given elevated (≥90th percentile of healthy volunteers) gas6 levels were estimated in regression models in the whole study population. After adjustment for age, sex, medications and comorbidity, subjects with elevated gas6 had an increased risk of VTE (OR of 16.3 (95% CI 5.8–45.7, P < 0.001) compared to those with lower levels of gas6. This association remains significant even among patients with a comparable age distribution. Among patients with VTE, mean gas levels showed a trend of higher levels in those with more extensive thrombi. There was no correlation between elevated gas6 levels and recurrent VTE. In conclusion, we demonstrate an association between VTE and elevated gas6 levels consistent with in vivo murine models of thrombosis. This constitutes a potential novel mechanism for thrombosis in humans and may aid in the understanding of the pathophysiology of VTE.     

Suppression of collagen‐induced arthritis in growth arrest and DNA damage–inducible protein 45β–deficient mice

Objective

Growth arrest and DNA damage–inducible protein 45β (GADD45β) is involved in stress responses, cell cycle regulation, and oncogenesis. Previous studies demonstrated that GADD45β deficiency exacerbates K/BxN serum–induced arthritis and experimental allergic encephalomyelitis (EAE) in mice, indicating that GADD45β plays a suppressive role in innate and adaptive immune responses. To further understand how GADD45β regulates autoimmunity, we evaluated collagen‐induced arthritis in GADD45β^–/– mice.

Methods

Wild‐type (WT) and GADD45β^–/– DBA/1 mice were immunized with bovine type II collagen (CII). Serum anticollagen antibody levels were quantified by enzyme‐linked immunosorbent assay. Expression of cytokines and matrix metalloproteinases in the joint and spleen was determined by quantitative polymerase chain reaction. The in vitro T cell cytokine response to CII was measured by multiplex analysis. CD4+CD25+ Treg cells and Th17 cells were quantified using flow cytometry.

Results

GADD45β^–/– mice showed significantly lower arthritis severity and joint destruction compared with WT mice. MMP‐3 and MMP‐13 expression was also markedly reduced in GADD45β^–/– mice. However, serum anti‐CII antibody levels were similar in both groups. FoxP3 and interleukin‐10 (IL‐10) expression was increased 2–3‐fold in splenocytes from arthritic GADD45β^–/– mice compared with those from WT mice. Flow cytometric analysis showed greater numbers of CD4+CD25+ Treg cells in the spleen of GADD45β^–/– mice than in the spleen of WT mice. In vitro studies showed that interferon‐γ and IL‐17 production by T cells was significantly decreased in GADD45β^–/– mice.

Conclusion

Unlike passive K/BxN arthritis and EAE, GADD45β deficiency in CIA was associated with lower arthritis severity, elevated IL‐10 expression, decreased IL‐17 production, and increased numbers of Treg cells. The data suggest that GADD45β plays a complex role in regulating adaptive immunity and, depending on the model, either enhances or suppresses inflammation.

     

Report on ocular biometry of microphthalmos, retinal dystrophy, flash electroretinography, ocular coherence tomography, genetic analysis and the surgical challenge of entropion correction in a rare case of Hallermann–Streiff–Francois syndrome

Purpose

To report new aspects of the phenotype including Retinal dystrophy and surgical challenges in Hallermann-Streiff Francois syndrome (HSFS).

Methods

Detailed phenotype of a female with HSFS was evaluated including skeletal changes, comprehensive eye examination, detailed ocular biometry, electroretinography and macular Ocular coherence tomography. Surgical notes of lid surgery for entropion were reviewed. Genetic screening was also done.

Results

Unique Ocular biometry with electroretinography changes, macular folds and fundus changes suggestive of an unreported Retinal dystrophy in a typical patient with HSFS were noted. Surgery was challenging both due to difficulty in endotracheal intubation anaesthesia because of the dento-facial abnormalities and the skin fragility.

Conclusion

This report provides additional information especially pigmentary retinal dystrophy, macular folds and electroretinography in HSFS. The microphthalmos had overlapping posterior segment findings usually reported with Nanophthalmos and Posterior microphthalmos. The surgical difficulties and outcomes of the rarely encountered adnexal abnormalities emphasize the need for a multi disciplinary approach for appropriate management.     

Favourable modulation of the inflammatory changes in hypercholesterolemic atherogenesis by a low-molecular-weight heparin derivative

BACKGROUND: In the hypercholesterolemic state, the net result of combined oxidative and nitrosative stress is a pro-inflammatory phenotype that is manifested as increased adhesion molecule expression, enhanced leucocyte trafficking, and increased vascular permeability. The present work explores the inflammatory aspects of hypercholesterolemic atherogenesis, and also evaluates the role of a low-molecular-weight heparin derivative (LMWH), Certoparin, on a biochemical basis. METHODS AND RESULTS: Two groups of male Wistar rats were fed an atherogenic diet (normal rat chow plus 4% cholesterol, 1% cholic acid and 0.5% thiouracil-CCT diet) for 2 weeks. While one was left untreated, the other was administered LMWH (300 microg/day/rat commencing on day 8 and continued for a week). Increased concentrations of plasma C-reactive protein and fibrinogen and cardiac TNF-alpha indicated severe inflammation in the atherogenic diet fed rats. In comparison, these biochemical indices of inflammation diminished significantly in the LMWH treated group (p < 0.001). Significant depletion of thiols, along with accentuated activities of the glutathione metabolising was observed in the cardiac and hepatic tissues of the untreated atherogenic rats, indicating heightened oxidative response. Tissue damage was marked by elevated levels of plasma and tissue hexose, hexosamine, hexuronic acid and sialic acid, which were reversed towards normalcy on LMWH administration. The activities of lysosomal enzymes (N-acetyl glucosaminidase, beta-glucuronidase, beta-galactosaminidase and cathepsin-D) showed a marked increase in the CCT-diet fed rats, while LMWH treated rats showed normal activities (p < 0.001). The osmotic fragility test revealed that the untreated hyperlipidemic rat erythrocytes were significantly fragile at high salt concentrations, while the response was normalized in the LMWH treated group (p < 0.05). Further, hypercholesterolemia induced downregulation of physiological nitric oxide levels was corrected upon treatment with heparin-derivative. CONCLUSION: The results of this work highlight the inflammatory changes in atherogenic conditions and that the low-molecular-weight heparin derivative affords substantial protection.     

Cholecystoappendicostomy for progressive familial intrahepatic cholestasis

We report a rare case of progressive familial intrahepatic cholestasis type 2 from India. The diagnosis was confirmed on the basis of gene mutation analysis. The child had intense pruritus refractory to conventional medical management. As liver biopsy did not reveal any cirrhosis, partial external biliary diversion was considered as an alternative to liver transplant. We performed cholecystoappendicostomy rather than the conventional method of using an ileal loop as a conduit between the gall bladder and abdominal wall. Child recovered completely.