The presence of messenger RNA for HLA class I in human platelets and its capability for protein biosynthesis

Summary. In order to determine whether platelets contain specific messenger RNA encoding for HLA class I molecules, polymerase chain reaction (PCR) was performed with RNA from different platelet donors. Two amplified 300 bp and 279 bp cDNA fragments were obtained which encompassed sequences from 321 to 620 and from 795 to 1073. The 300 bp fragment encodes exon 2 and exon 3, the 279 bp encodes a portion of exon 4, exon 5, exon 6 and a portion of exon 7. A 300 bp nested PCR product from one donor, that encoded for the highly polymorphic region α2, was cloned and sequenced. The resulting nucleotide sequences fitted to the expected sequence for HLA B*3801 of this donor. Sequence analysis of the 279 bp PCR product demonstrated the presence of exon 5 encoding for the 117 bp transmembrane domain.
In addition, de novo protein biosynthesis was studied by radioimmunoprecipitation of HLA class I molecules from ³⁵S-methionine metabolically labelled platelet lysates with a monoclonal antibody (mab) w6/32 specific for a monomorphic epitope on the heavy chain of HLA class I antigens. Analysis of the immunoprecipitates on SDS polyacrylamide gel electrophoresis showed a specific band with apparent molecular weight (M_r) of 44 kD corresponding to integral membrane HLA protein.
On the basis of these results, we conclude that platelets contain specific messenger RNA encoding for HLA class I molecules and have the capability to synthesize the integral HLA membrane protein.     

Malignancy may adversely influence the quality and behaviour of oocytes

A case series of eight cycles of in-vitro fertilization (IVF) in five women diagnosed with malignant disorders is presented. These patients chose to defer definitive treatment for a chance for preservation of potential fertility. The response of these patients to ovarian stimulation, and the outcome, was compared with 17 IVF cycles in 12 age- matched patients with isolated tubal infertility. An apparent adverse influence of malignant disease on the quality and behaviour of oocytes was observed. Despite a comparable total number of oocytes per cycle in the two groups, a significantly reduced percentage of mature oocytes was retrieved per cycle from patients with malignant diseases. The oocytes from patients with malignant disorders were of a poorer quality and exhibited a significantly impaired fertilization rate compared to the controls. We propose that neoplastic processes, irrespective of the site or cell of origin, may have a detrimental impact on the biology of oocytes, an effect akin to that seen on spermatozoa in men with certain malignancies.      

Microglial inflammation in the parkinsonian substantia nigra: relationship to alpha-synuclein deposition

Background  

The role of both microglial activation and alpha-synuclein deposition in Parkinson's disease remain unclear. We have tested the hypothesis that if microglia play a primary role in Parkinson's disease pathogenesis, the microglial "activated" phenotype should be associated with histopathological and/or clinical features of the disease.     

Overall nutritional completeness (ONC): A new integrative measure of nutritional and agricultural performance quality of garden crops

In spite of increased efforts to integrate nutrition and agriculture there are relatively few methods developed to effectively do so in the design and analysis of agricultural interventions. In order to develop and test a method to link agricultural production and nutritional benefits, fall gardens were planted by researchers and low‐income households in five locations in the New River Valley, Virginia. Edible portion yield per square meter was calculated as a basic nutrition/agricultural yield measure. In order to use a more discriminating measure, crop nutrient yield equivalents (CNYE) for limiting nutrients (vitamins A and C, iron, and calcium) were calculated. The CNYE indicates the number of days that 100% of the USRDA would be supplied. Summing the CNYE scores across nutrients provided a ranking of the overall nutritional completeness (ONC) of the crops. The ONC ranking thus supplies a measure of yield and nutritional efficiency that can be adapted for different nutrition requirements and gardening conditions.     

A systematic review of patient‐reported outcomes in patients with cutaneous lupus erythematosus

Cutaneous lupus erythematosus (CLE) is a chronic dermatological autoimmune disease marked by photosensitive lesions that can lead to hyperpigmentation changes, scarring and hair loss. Health‐related quality of life (HRQoL) in patients with CLE is severely impaired. Given the heterogeneous nature of CLE, health perceptions of patients can differ significantly from those of clinicians. It is important to use subjective measures, such as patient‐reported outcomes (PROs), to capture HRQoL data in patients with CLE. We conducted a systematic review of published PRO instruments used in measuring HRQoL in patients with CLE. Also, we examined the disease burden on HRQoL in patients with CLE. To identify studies, PubMed/MEDLINE, Web of Science and CINAHL were searched using ‘CLE/cutaneous lupus erythematosus’ in combination with PRO‐related keywords such as ‘quality of life’, ‘self‐report’ and ‘instrument’. English‐language articles published between 2003 and 2014 were identified. A total of 482 citations were identified in the initial search. Eleven studies met our inclusion criteria, and five PRO instruments were found to be used: Skindex (versions 16 and 29), Dermatology Life Quality Index, 36‐Item Short‐Form Health Survey, and visual analogue scales for pain and pruritus. Patients with CLE reported having poor quality of life and experienced symptoms ranging from pain, pruritus and fatigue to photosensitivity. There is a limited number of studies examining PRO in patients with CLE. While our findings suggest that quality of life in patients with CLE is poor, further studies are needed to understand better the impact of CLE from patients’ perspectives.     

Endotoxin-induced suppression of erythropoiesis: the role of erythropoietin and a heme synthesis stimulating factor

The regulation of erythropoiesis is primarily controlled by erythropoietin (Ep). Recently, however, other factors that both stimulate and inhibit erythropoiesis have been reported. Using an in vitro liquid culture of bone marrow cells, a factor in normal mouse serum was demonstrated that markedly stimulated heme synthesis by marrow erythroid cells. In this study, the role of this heme synthesis stimulating factor (HSF) and Ep in the erythropoietic suppression caused by endotoxin administration to mice was examined. Although HSF levels did not alter appreciably after endotoxin injection, marrow erythroid cells from these animals became unresponsive to the factor. This could be reversed if Ep was added to the culture in vitro or if the hormone was injected into the mice 18 hr prior to harvesting the marrow. This marrow erythroid cell response is identical to that seen in animals in whom Ep levels are markedly reduced, such as that found in exhypoxic polycythemia, and suggest a decrease in the hormone following endotoxin administration. Additional studies demonstrated that when Ep was injected into mice 6 hr after endotoxin administration, an increase in femoral erythroid colony-forming units (CFU-E), proerythroblast number, and 59 Fe incorporation into femoral marrow cells could be demonstrated. These findings, together with the marrow erythroid cell response to the hormone, suggest that the mechanism for suppression of erythropoiesis after endotoxin injection is a reduction in the level of circulating Ep.     

Liver steatosis assessment: Correlations among pathology,radiology, clinical data and automated image analysis software

Quantitating hepatic steatosis is important in many liver diseases and liver transplantation. Since steatosis estimation by pathologists has inherent intra- and inter-observer variability, we compared and contrasted computerized techniques with magnetic resonance imaging measurements, pathologist visual scoring, and clinical parameters. Computerized methods applied to whole slide images included a commercial positive pixel count algorithm and a custom algorithm programmed at our institution. For all liver samples (n = 59), including pediatric, adult, frozen section, and permanent specimens, statistically significant correlations were observed between pathology, radiology, and each image analysis modality (r = 0.75–0.97, p < 0.0001), with the strongest correlations in the pediatric cohort. Statistically significant relationships were observed between each method and with body mass index (r = 0.37–0.56, p from <0.0001 to <0.05) and with albumin (r = 0.55–0.64, p < 0.05) but not with alanine aminotransferase or aspartate aminotransferase. Although pathologist assessments correlated (r = 0.64–0.86, 0.92–0.97, and 0.78–0.91 for microvesicular, macrovesicular, and overall steatosis, respectively), the absolute values of hepatic steatosis visual assessment were susceptible to intra- and inter-observer variability, particularly for microvesicular steatosis. Image analysis, pathologist assessments, radiology measurements, and several clinical parameters all showed correlations in this study, providing evidence for the utility of each method in different clinical and research settings.     

Electrocardiographic predictors of out-of-hospital sudden cardiac arrest in patients with coronary artery disease

Sudden cardiac arrest (SCA), due mainly to coronary artery disease (CAD), is a leading cause of death. To identify electrocardiographic and clinical differences between patients with CAD with and without SCA, 87 victims of SCA with CAD were compared with 131 patients with CAD without SCA. Patients' latest routine electrocardiograms and clinical variables were compared. Patients with CAD with and without previous myocardial infarctions (MIs) were included. Patients with SCA had a higher incidence of echocardiographic evidence of left ventricular hypertrophy and/or heart failure than controls. The median left ventricular ejection fractions for patients with SCA with and without previous MIs were 0.30 (interquartile range 0.24 to 0.41) and 0.41 (interquartile range 0.25 to 0.56). The median time between the last electrocardiographic assessment and SCA was 59 days (interquartile range 29 to 137). Regarding electrocardiographic characteristics, in patients with and without previous MIs, QRS width (odds ratio 1.032, 95% confidence interval 1.012 to 1.053, p = 0.002, and odds ratio 1.035, 95% confidence interval 1.015 to 1.056, p = 0.001) was the only significant predictor of SCA. In conclusion, in patients with CAD, regardless of a previous MI, a longer QRS width and echocardiographic parameters consistent with heart failure are associated with SCA, even in patients with ischemic cardiomyopathy currently not eligible for an implantable cardioverter-defibrillator.