Optimization of a fibrin scaffold for sustained release of an adenoviral gene vector

One of the major limitations of adenovirus as an efficient gene delivery vehicle is its transient nature of transgene expression. At present, no biodegradable release system exists that permits the sustained release of gene delivery. This study aimed to optimize a biodegradable fibrin scaffold for sustained adenoviral mediated gene delivery. Scaffold formulations were fabricated incorporating an adenoviral vector encoding beta-galactosidase, and each scaffold was incubated in elution fluid. The level of transfection of cells was measured to assess virus diffusion from each scaffold at different time periods. Results showed that viral particles were present in the elution fluid and had the ability to transfect cells at different time points. Depending on the scaffold formulation, variable rates of diffusion were seen. Analysis of scaffold microarchitecture using stereo pairs showed that variable pore depth is seen in different fibrin scaffold concentration, which may be a determining factor in the diffusion rate. It was concluded that sustained release of the adenovirus could be attained up to 192 h at the optimal concentration of 60 mg/mL fibrinogen and 4 IU thrombin.     

Chronic morphine exposure causes pronounced virus replication in cerebral compartment and accelerated onset of AIDS in SIV/SHIV-infected Indian rhesus macaques

Six morphine-exposed and 3 control male Indian rhesus macaques were intravenously inoculated with mixture of SHIV(KU), SHIV(89.6)P and SIV/17E-Fr. These animals were followed for a period of 56 weeks in order to determine CD4 and CD8 profile, viral loads in plasma and cerebrospinal fluid (CSF), relative distribution of 3 pathogenic viruses in blood and brain, binding as well neutralizing antibody levels and cellular immune responses. Both morphine-exposed and control macaques showed a precipitous loss of CD4+ T cells; control animals, however, showed a greater tendency to recover these cells than did their morphine-exposed counterparts. The plasma and CSF viral loads were significantly higher in morphine-exposed group than those in the control group. Four morphine-exposed animals succumbed to SIV/SHIV-induced AIDS at week 18, 19, 20 and 51; post-infection with neurological disorders was found in 3 of the 4 animals. At the end of the 56-week observation period, 2 morphine-exposed and 3 control animals were still alive. All 3 viruses replicated in the blood of both morphine-exposed and control macaques, but the cerebral compartment showed a selection phenomenon; only SIV/17E-Fr and SHIV(KU) successfully crossed the blood brain barrier (BBB). The morphine-exposed macaques further favored viral migration through the blood brain barrier (BBB). SIV/17E-Fr crossed the BBB within 2 weeks in both morphine-exposed and control macaques, whereas SHIV(KU) crossed the BBB more rapidly in morphine-exposed than in control macaques. Three morphine-exposed macaques (euthanized at weeks 18, 19 and 20) did not develop cellular or humoral immune responses, whereas the other 3 morphine-exposed and 3 control macaques developed both cellular and humoral immune responses.     

Urothelial carcinoma with rhabdoid features: report of 6 cases

Extrarenal rhabdoid tumors have been described in a variety of primary sites with only rare case reports of urothelial carcinomas with rhabdoid features in the literature. In this report, we describe the clinicopathologic characteristics, including clinical follow-up on 6 cases of urothelial carcinoma with prominent rhabdoid features. Four cases were retrieved from the consultation files of one of the authors and 2 were retrieved from the surgical pathology files at our institution. The patients were all men, with ages ranging from 53 to 86 years (mean, 66.5 years). Patients initially presented with hematuria or obstructive symptoms. The sites included bladder (n = 4) and renal pelvis (n = 2). All cases had a prominent rhabdoid component (mean, 60%), ranging from 40% to 80%. In addition to the rhabdoid component, multiple coexistent histological components were seen, including in situ urothelial carcinoma (carcinoma in situ) and high-grade papillary urothelial carcinoma (n = 2), poorly differentiated carcinoma with small-cell features (n = 1), sarcomatoid (n = 2), and a myxoid component (n = 2). All cases in this series had focal or diffuse positive staining with one or more cytokeratin markers (epithelial membrane antigen, CAM 5.2, AE1/AE3). Of the 6 patients, 4 were treated initially with surgery (radical cystoprostatectomy, n = 2; radical nephrectomy, n = 2). Of 6 patients, 2 died within 1 month, whereas a third patient died within 4 months. The remaining 3 patients were alive at 3, 3, and 9 months after diagnosis. The histological and immunohistochemical findings in this study serve to broaden the morphological spectrum of urothelial carcinomas with prominent rhabdoid features and add further evidence as to their poor prognosis.     

Chronic liver disease related to 6-thioguanine in children with acute lymphoblastic leukaemia

BACKGROUND/AIMS: The United Kingdom (UK) acute lymphoblastic leukaemia (ALL) 97/99 clinical trial compared 6-mercaptopurine (6MP) with 6-thioguanine (6TG) as maintenance therapy for childhood ALL. Review of interim results has led to discontinuation of the 6TG arm. METHODS: We report six children with ALL, who presented with splenomegaly after a median (range) treatment duration of 12 (6-22) months. All these children were treated in the 6TG-arm. RESULTS: The median (range) age at presentation was 6.6 (3.2-11.5) years. There were five boys. The presenting features were splenomegaly in all and hepatomegaly in four. AST was abnormal in one (80 IU/l, normal range 10-50). Abdominal sonography showed an altered texture of the liver parenchyma and confirmed splenomegaly. Microscopy showed findings within the spectrum of occlusive venopathy and nodular regenerative hyperplasia (NRH). After a median (range) follow-up of 23 (4-36) months splenomegaly and thrombocytopenia, suggestive of progressive portal hypertension, continue to worsen in all children. CONCLUSIONS: 6TG is associated with chronic hepatic toxicity and progressive portal hypertension on follow-up. Microscopy showed NRH in all patients with features in keeping with an intrahepatic occlusive venopathy and variable parenchymal atrophy and loss.     

Acetaminophen-induced Hepato- and Nephrotoxicity and Amelioration by Silymarin and Terminalia chebula in Rats

Experimental study was conducted to evaluate the hepato- and renoprotective effect of silymarin and Terminalia chebula against experimentally-induced acetaminophen (APAP) toxicity in rats. Oral administration of APAP @ 500 mg/kg for 1 to 3 days to all the four groups (six rats in each) resulted in significant elevation of serum triglycerides, total cholesterol, blood urea nitrogen, serum creatinine, and aspartate transaminase activity. Post-treatment with silymarin @ 25 mg/kg and T. chebula 125 mg/kg in groups 2 and 3 and their combination to group 4 from day 4 to 14 has significantly reversed the alterations of above said markers and offered better protection. The results of the study enunciated that silymarin and T. chebula exhibit good hepato- and nephro-protection against APAP toxicity.     

Lichen sclerosus of lips: a clinical and histopathologic study of 27 cases

Background & Methods Oral lichen sclerosus (LS) has been considered uncommon and involvement of lips extremely rare. We reviewed the clinical and histologic features of 72 cases of LS with oral/genital involvement, seen in our institute from 2002 to 2007. Results Lichen sclerosus was diagnosed with exclusive genital lesions in 45, exclusive lip involvement in 20, and orogenital involvement in seven cases. Fifteen of 27 histologically confirmed lip LS lesions were considered as early inflammatory or presclerotic, eight were intermediate/progressive, and four as late resolved lesions. Lip LS presented as asymptomatic vitiligoid lesions in 70% and dermal sclerosis was demonstrable in only 44%, which was limited to the papillary layer. This was in contrast with genital LS lesions which were asymptomatic in only 12% and demonstrated both papillary and reticular dermal sclerosis in 69%. Conclusion Lip LS is far less symptomatic and destructive with limited dermal sclerosis compared with genital LS. Greater awareness and histologic assessment are essential for diagnosis because of the misleading vitiligoid appearance. “Vitiligoid LS” a superficial variant proposed by Borda can be aptly applied to lip LS. Dermatologists need to be aware of this rarely reported manifestation of LS as it adds to the spectrum of oral lichenoid lesions and lichenoid dysplasia, which are suspected to have a malignant potential.     

The relationship between body composition and bone mineral density in postmenopausal Turkish women

In a retrospective cross-sectional study among 202 postmenopausal women aged 46–75 years, we aimed to investigate the relationship between body composition and bone mineral density (BMD) to determine whether fat mass or lean mass is a better determinant of BMD in Turkish postmenopausal women. Lumbar spine (L1–L4) and proximal femur BMD were measured by dual energy X-ray absorbsiometry. Body composition analysis was performed by bioelectric impedance method and fat mass, lean mass, and percent fat were measured. Both fat mass and lean mass were positively correlated with BMD at the lumbar spine and proximal femur, weight and body mass index. Lean mass was also positively correlated with height and negatively correlated with age and years since menopause (P < 0.01). The correlations of fat mass and lean mass with BMD at the lumbar spine and proximal femur remained significant after adjustment for age, years since menopause and height. When the lean mass was adjusted together with age, years since menopause and height, the significant relationship between the fat mass and BMD continued, however the significant correlation between the lean mass and BMD disappeared at all sites after adjustment for fat mass. In multiple regression analyses, fat mass was the significant determinant of all BMD sites. Our data suggest that fat mass is the significant determinant of BMD at the lumbar spine and proximal femur, and lean mass does not have an impact on BMD when fat mass was taken into account in Turkish postmenopausal women.     

Rates and causes of death from non-ST elevation acute coronary syndromes: Ten year follow-up of the PRAIS-UK registry

Background

Long term nationally representative mortality rates following acute coronary syndrome (ACS) admissions are lacking beyond 5 years. We report rates and causes of mortality at approximately 10 years from PRAIS-UK.

Methods

PRAIS-UK was a prospective registry of 1046 non-ST-elevation ACS admissions to 56 UK hospitals between 1998 and 1999. 493 patients surviving to 6 months were consented to long term follow-up. We identified deaths and causes (ICD codes) via the UK central death register and examined the influence of baseline characteristics and early revascularisation procedures. A modified GRACE risk score was constructed to determine the association of baseline score with long term risk of death.

Results

The mean age was 66 years and 40% were women. After a median follow-up of 11.6 years (IQR 6.3–11.9), 46% (225) of patients had died with 55% being classified as cardiovascular. In a multivariate analysis, the following variables were associated with higher mortality (hazard ratio [HR] and 95% confidence intervals [CI]): age (10 years increase) 2.14 (1.87 to 2.45), ST depression or bundle branch block (compared to normal ECG) 1.68 (1.06 to 2.67), and history of heart failure (compared to no HF) 1.81 (1.28 to 2.56). The HR for risk of death in patients who received a revascularisation procedure (versus those who did not) in the first 6 months was 0.41 (0.24 to 0.69). The mean adapted GRACE score was 99.3 ± 26.4, associated with approximately 50% mortality at 10 years.

Conclusions

Non-ST elevation ACS is associated with about 50% mortality over 10 years that may be improved by early revascularisation. Well designed long-term registries can provide key data to determine prognosis and burden of disease.     

Design,Synthesis, and Biological Evaluation of Catecholamine Vehicle for Studying Dopaminergic System

Catecholamine mimetic EDTA‐bis(tyramide) was synthesized and characterized by various spectroscopic techniques (NMR, mass spectroscopy) and λ_em 310 nm for the excitation at 270 nm. Molecular docking studies were performed with human serum albumin (PDB 1E78), showing binding pattern with amino acid residues Arg218, Arg222, and Lys444, identifies the ligand‐human serum albumin interaction for the transportation affinity of the ligand at the specific site of the target. Subsequently, binding study with human serum albumin at λ_ex = 350 nm found to be 5.847 × 10⁴ m ^?1 shows effective quenching effect. Additionally, to go more insight, acetylcholinesterase binding affinity was investigated, which shows 90% binding affinity for the 10 mm concentration. IC50 value was found 18.60 μm for MAO‐B inhibition. Finally, EDTA‐bis(tyramide) labeled with ^99mTc to investigate its in vivo radiopharmaceutical efficiency having 97% binding affinity with 98% radiochemical purity. In vivo studies were carried out for ^99mTc‐EDTA‐bis(tyramide) included blood kinetics showed a quick wash out from the circulation via renal route, and biodistribution revealed that maximum %ID/g was found in kidney at 1 h, and its scintigraphy image shows 3.96% brain uptake with respect to whole body.