Association of human polyomavirus JC with peripheral blood of immunoimpaired and healthy individuals

JC virus (JCV) infection is regularly asymptomatic in healthy individuals. In contrast, in immunocompromised individuals, highly activated virus replication may lead to PML. Peripheral blood cells (PBCs) are found to habor JCV DNA in healthy and diseased individuals and it is discussed that they might be responsible for dissemination of the virus to the central nervous system (CNS) during persistence. To better understand the role of JCV DNA in PBCs for persistent infection and pathogenesis, the authors characterized the extent of JCV infection in Ficoll-gradient purified blood cells (peripheral blood mononuclear cells [PBMCs]) of healthy and human immunodeficiency virus type 1 (HIV-1)-infected individuals. Virus activation in PBMCs from healthy JCV-infected individuals was found at a rate of 0% to 38% at low polymerase chain reaction (PCR) sensitivity. In progressive multifocal leukoencephalopathy (PML) patients, a stronger signal was found, indicating increased virus activation. JCV DNA was regularly detected in T and B lymphocytes and in monocytes at low levels. However, granulocytes were shown to be the predominant reservoir of JCV DNA harboring high copy numbers. Although the overall distribution of viral genomes holds true for the population studied, in the individual, a markedly changed pattern of distribution can be found.     

Screening for diabetes: sensitivity and positive predictive value of risk factor total.

OBJECTIVE: Screening for diabetes is recommended for individuals > or =45 years of age, or earlier if they manifest > or = one specific risk factors. This study examined the sensitivity and positive predictive value (PPV) of risk factor total for identifying individuals with diabetes and prediabetes. DESIGN: Subjects were interviewed to assess the presence of risk factors. Fasting plasma glucose levels were obtained. SETTING: The study occurred at a health fair in Greensburg, PA. PATIENTS: Six hundred sixty-one Caucasians between the ages of 19 and 100. RESULTS: Using the criterion of screening individuals with > or = one risk factors detected 100% of both diabetics and prediabetics. This dropped to 91.2% when screening individuals with > or = two factors. The PPV of the risk factor total was poor (80% of individuals with a total of four factors were not diabetic). The ability of the risk factor total to predict individuals with impaired glucose metabolism (prediabetics + diabetics) was considerably better, and increased almost linearly with the risk factor total. Of the subjects with normal glucose values, the mean glucose increased as the risk factor total increased. CONCLUSION: While the sensitivity of using > one risk factor as an algorithm to screen is 100% for identifying diabetics, the PPV of risk factor analysis for identifying diabetics is poor. The same algorithm works well to identify at-risk individuals, presumably allowing early intervention and education.     

Analysis of five specific scores for cervical spondylogenic myelopathy

The ability to compare various results that measure clinical deficits and outcome is a necessity for successful worldwide discussion about cervical spondylogenic myelopathy (CSM) and its treatment. There is hardly any information in literature how to value and compare outcome assessed by different scores. In a retrospective study we objectively evaluated the Nurick-score, Japanese-orthopaedic-association-score (JOA-Score), Cooper-myelopathy-scale (CMS), Prolo-score and European-myelopathy-score (EMS) using the data of 43 patients, all of whom showed clinical and morphological signs of CSM and underwent operative decompression. The scores were assessed pre- and postoperatively. The correlation between the score-results, anamnesis, clinical and diagnostic data was investigated. All the scores show a statistically significant correlation and measure postoperative improvement. With exception of the Prolo-score all scores reflect clinical deficits of CSM. The Prolo-score rates the severity of CSM on the state of the economic situation above clinical symptoms. The main differences of the scores are shown in the number of patients showing postoperative improvement, varying between 33% (Nurick-score) and 81% (JOA-score). The recovery-rates, as a measure of the cumulative improvement of all the symptoms, show less variation (23–37%). The differences of the recovery-rate were only statistically significant between JOA-score, Nurick-score and EMS (P < 0.05), whereas all the other scores showed no significant differences. To assess the postoperative successes, the evaluation of the recovery-rate is essential. There is no significant difference in the recovery-rate amongst the majority of the scores, which allows a good comparison of the results from different studies. Nevertheless, it is always important to differentiate the therapy results of CSM published worldwide.     

Statins, neuromuscular degenerative disease and an amyotrophic lateral sclerosis-like syndrome: an analysis of individual case safety reports from vigibase.

BACKGROUND: The WHO Foundation Collaborating Centre for International Drug Monitoring (Uppsala Monitoring Centre [UMC]) has received many individual case safety reports (ICSRs) associating HMG-CoA reductase inhibitor drug (statin) use with the occurrence of muscle damage, including rhabdomyolysis, and also peripheral neuropathy. A new signal has now appeared of disproportionally high reporting of upper motor neurone lesions. AIM AND SCOPE: The aim of this paper is to present the upper motor neurone lesion cases, with other evidence, as a signal of a relationship between statins and an amyotrophic lateral sclerosis (ALS)-like syndrome. The paper also presents some arguments for considering that a spectrum of severe neuromuscular damage may be associated with statin use, albeit rarely. The paper does not do more than raise the signal for further work and analysis of what must be regarded as a potentially very serious and perhaps avoidable or reversible adverse reaction, though it also suggests action to be taken if an ALS-like syndrome should occur in a patient using statins. METHODS: The 43 reports accounting for the disproportional reports in Vigibase (the database of the WHO Programme for International Drug Monitoring) are summarised and analysed for the diagnosis of an ALS-like syndrome. The issues of data quality and potential reporting bias are considered. RESULTS: 'Upper motor neurone lesion' is a rare adverse event reported in relationship to drugs in Vigibase (a database containing nearly 4 million ICSRs). Of the total of 172 ICSRs on this reported term, 43 were related to statins, of which 40 were considered further: all but one case was reported as ALS. In 34/40 reports a statin was the sole reported suspected drug. The diagnostic criteria were variable, and seven of the statin cases also had features of peripheral neuropathy. Of a total of 5534 ICSRs of peripheral neuropathy related to any drug in Vigibase, 547 were on statins. The disproportional reporting of statins and upper motor neurone lesion persisted after age stratification, and such disproportionality was not seen for statins and Parkinson's disease, Alzheimer's disease, extrapyramidal disorders, or multiple sclerosis-like syndromes. DISCUSSION: Because the cases were sometimes atypical we propose the use of the term 'ALS-like syndrome' and speculate whether this is part of a spectrum of rare neuromuscular damage. The diagnosis of ALS is often problematic, and the insidiousness and chronicity of the disease make causality with a drug difficult to assess. The disproportionally high reporting makes this an important signal nevertheless, since ALS is serious clinically and statins are so widely used. Wide use of the statins also makes a chance finding more probable, but is unlikely to cause disproportional reporting when there are no obvious biases identified. CONCLUSION: We emphasise the rarity of this possible association, and also the need for further study to establish whether a causal relationship exists. We do advocate that trial discontinuation of a statin should be considered in patients with serious neuromuscular disease such as the ALS-like syndrome, given the poor prognosis and a possibility that progression of the disease may be halted or even reversed.     

Hypocalcemic symptoms during plateletpheresis using the COBE Spectra: A comparison of oral combination of 600mg calcium+300mg magnesium+100IU vitamin D3 vs. a 1000mg calcium in symptomatic donors.

BACKGROUND: The aim of this study was to find an effective treatment for hypocalcemic symptoms during plateletpheresis and to evaluate if a combination of calcium, magnesium and vitamin D3 is more effective in comparison to routine calcium supplementation. MATERIAL AND METHODS: A study group consisting of 10 donors, having a history of previous hypocalcemic symptoms during plateletpheresis, donated platelets twice in a one-month period. During the first donation combination tablets (600mg Ca+300mg Mg+100IU vitamin D3) were used to treat hypocalcemic symptoms while routine treatment calcium carbonate tablets (1000mg Ca) were used during the second donation. If symptoms persisted after 10min the same dose was repeated. A control group, with no supplementation, consisting of five donors, with no history of hypocalcemic symptoms, were included. Donor subjective symptoms were graded and recorded on four occasions: at the start of plateletpheresis, when symptoms appeared, 10min after the first tablet and at the end of donation. Samples for analysis of ionized calcium (iCa), magnesium and potassium were also taken at the same occasions. RESULTS: All donors from the study group experienced minor or medium hypocalcemic symptoms and needed a second dose of supplementation. Calcium carbonate tablets completely relieved the hypocalcemic symptoms in six donors, it had no effect on three donors and one donor experienced aggravated symptoms. The combination tablets completely relieved the symptoms in three donors, one donor experienced a partial relief and six donors had no relief of symptoms. There were no significant differences in iCa, potassium and magnesium levels were noted in the study group irrespective of which tablets were used for treatment of hypocalcemic symptoms. After plateletpheresis the median iCa levels declined by 30% and potassium levels declined by 3-11% in all donors while the magnesium levels were not significantly affected. There was no correlation between the presence of symptoms and the changed levels of iCa or magnesium. CONCLUSION: Addition of magnesium and vitamin D3 to calcium seems to have no beneficial effect in the treatment of hypocalcemic symptoms in plateletpheresis donors.     

Impairment of bone turnover in elderly women with hip fracture

Summary Hip fracture is one of the most severe consequences of osteoporosis affecting aged women. However, abnormalities of bone turnover responsible for bone loss in this condition have not been clearly defined. To further evaluate the bone metabolic status of women sustaining hip fracture, we have prospectively measured serum osteocalcin as a marker of bone formation and urinary excretion of pyridinoline (Pyr) and deoxypyridinoline (D-pyr) cross-links as markers of bone collagen degradation in 174 independently living women (80 ± 8 years) within a few hours after a hip fracture. Comparison was made with 77 age-matched controls (80 ± 5 years) and 17 premenopausal women (39 ± 3 years). In addition 15 of the patients were followed with daily measurements during the first postoperative week. At the time of admission osteocalcin was 20% lower in the fractured women compared to the elderly controls (7.6 ± 3.8 vs. 9.5 ± 4.5 nglml,P = 0.001). Pyr and D-pyr were 36% and 40% higher, respectively (P = 0.0001), than in elderly controls and 85% and 76% higher than in premenopausal controls (P = 0.0001). Serum osteocalcin did not correlate with the cortisol level measured at the same time (r = 0.03, ns), nor with serum albumin and creatinine. Serum osteocalcin remained unchanged within 18 hours after fracture, whereafter it progressively decreased until the third postoperative day. No correlation was noted between the excretion of pyridinoline cross-links and the time elapsed from fracture.These data suggest that the abnormal levels of osteocalcin and pyridinolines are unrelated to traumatically induced acute changes, but reflect abnormalities of bone turnover existing prior to the fracture. Thus, hip-fracture patients have biochemical evidence of decreased bone formation and increased bone resorption when compared to age-matched controls. We suggest that these abnormalities may play a role in the decrease of the bone mass and the consequently increased bone fragility that characterize the osteoporotic hip fracture in the elderly.     

Possible Relation of p53 and mdm-2 Oncoprotein Expression in Thyroid Carcinoma: A Molecular-Pathological and Immunohistochemical Study on Paraffin-Embedded Tissue

Routinely processed tissues from a series of benign and malignant thyroid lesions were immunohistochemically investigated with antibodies against p53 and mdm-2. p53 was immunolocalized in <10% of nuclei in 2/80 nodular goiters, 2/60 follicular adenomas, 26/68 follicular carcinomas, 7/40 papillary carcinomas, 3/10 “insular” carcinomas, and 10/31 anaplastic carcinomas. More than 10% positively stained nuclei were found in 2 widely invasive follicular, 2 insular, and 15 anaplastic carcinomas. All p53-positive cases showed a concomitant immunohistochemical mdm-2 expression; an immunohistochemical colocalization on serial section was demonstrated in 12 anaplastic carcinomas. Screening by polymerase chain reaction single-strand conformation polymorphism (PCR-SSCP) analysis of these 12 cases revealed no relevant mutations in the coding regions of exons 2–11 of the p53 gene. Additionally, 1 follicular adenoma, 6 follicular carcinomas (4 minimally and 2 widely invasive), 1 papillary, and 2 poorly differentiated insular carcinomas were mdm-2 positive without immunohistochemically detectable p53 expression. These results provide evidence that wild-type p53 expression in thyroid carcinomas may be associated with mdm-2 induced formation of stable complexes. However, the role of p53 mutations and p53 protein inactivation owing to other factors (e.g., mdm-2) in the progression of thyroid carcinomas is still poorly understood.     

Analysis of cytogenetic alterations in stage III serous ovarian adenocarcinoma reveals a heterogeneous group regarding survival, surgical outcome, and substage

Ovarian cancer is the leading cause of death among patients with gynecological cancers, but the biology of these tumors is still among the least understood of all major human malignancies. In this study, comparative genomic hybridization was used to determine chromosomal alterations in 98 stage III serous papillary adenocarcinomas. The tumors were grouped according to survival and the main prognostic factors stage and surgical outcome. There were chromosomal imbalances that were significantly more common in tumors from patients who died than in tumors from patients who survived: gains of 1q24-qter and losses of 4p, 4q31.1-qter, 5q12-q22, 8p, 16q, and X. Furthermore, we observed that gains of 8q23-8q24.2 and losses of 4p, 4q13-4q26, 4q31.1-qter, 5q12-q22, 8p, and 16q were significantly more common in tumors from patients with macroscopic residual tumor after primary surgery, compared to tumors from those who had undergone radical surgery. Gains of 3q13.3-qter, 6p, 7q21-q31, and 11q13-q23 and losses of 4q31.1-qter and 16q were more common in stage IIIc tumors than in stage IIIa+b tumors. On the basis of our results, we suggest that there are biological differences among the groups mentioned above and that absence of chromosomal aberrations in specific regions predicts a good clinical outcome for individual patients.     

Characterization of Virus-Responsive Plasmacytoid Dendritic Cells in the Rhesus Macaque

Plasmacytoid dendritic cells (PDC) are potent producers of alpha interferon (IFN-α) in response to enveloped viruses and provide a critical link between the innate and adaptive immune responses. Although the loss of peripheral blood PDC function and numbers has been linked to human immunodeficiency virus (HIV) progression in humans, a suitable animal model is needed to study the effects of immunodeficiency virus infection on PDC function. The rhesus macaque SIV model closely mimics human HIV infection, and recent studies have identified macaque PDC, potentially making the macaque a good model to study PDC regulation. In this study, we demonstrate that peripheral blood PDC from healthy macaques are both phenotypically and functionally similar to human PDC and that reagents used for human studies can be used to study macaque PDC. Both human and macaque PBMC expressed IFN-α in response to herpes simplex virus (HSV), the prototypical activator of PDC, as measured by using an IFN bioassay and IFN-α-specific enzyme-linked immunospot assays. Similar to human PDC, macaque PDC were identified by using flow cytometry as CD123⁺ HLA-DR⁺ lineage⁻ cells. In addition, like human PDC, macaque PDC expressed intracellular IFN-α, tumor necrosis factor alpha, macrophage inflammatory protein 1β/CCL4, and IFN-inducible protein 10/CXCL10 upon stimulation with HSV, all as determined by intracellular flow cytometry. We found that IFN regulatory factor 7, which is required for the expression of IFN-α genes, was, similar to human PDC, expressed at high levels in macaque PDC compared to monocytes and CD8⁺ T cells. These findings establish the phenotypic and functional similarity of human and macaque PDC and confirm the utility of tools developed for studying human PDC in this animal model.