Evidence for restricted replication of rubella virus in rat glial cells in culture

Intracellular rubella virus (RV) polypeptide synthesis during a productive infection of murine fibroblasts (L2 cells) has been investigated using immune precipitation techniques. Four structural and three additional intracellular polypeptides (p75, p60, VP44, VP41, p30, VP24, and VP19) were observed following polyacrylamide slab gel electrophoresis and autoradiography. However, when normal rat glial (RG) cells were infected only five polypeptides could be observed (p75, p60, VP44, VP41, and VP19). No infectious RV could be detected in tissue culture medium from flasks of infected RG cells. Calculations of relative concentrations of intracellular RV polypeptides precipitated from L2 and RG cells indicated that undetectable amounts of p30 and VP24, diminished amounts of p60 and VP19 and more than twice as much p75 were precipitated from infected RG cells. These data indicate that there is restricted replication of RV in normal rat glia.     

Association of DLG5 R30Q variant with inflammatory bowel disease

Crohn's disease (CD) and ulcerative colitis (UC) are chronic inflammatory diseases of the gastrointestinal system known as the inflammatory bowel diseases (IBD). Recently, Stoll and colleagues reported a novel finding of genetic variation in the DLG5 gene that is associated with IBD (CD and UC combined). We present here a study of the genetic variation described in that report in two well-powered, independent case-control cohorts and one family-based collection, and confirm the proposed association between IBD and the R30Q variant of DLG5 in two of the three studies. We are, however, unable to replicate the other proposed association to the common haplotype described in Stoll et al and suggest that this other finding could conceivably have been partially a statistical fluctuation and partially a result of LD with the replicated R30Q association. This study provides support for the hypothesis that DLG5 constitutes a true IBD risk factor of modest effect.     

Fulminant disseminated Varicella Zoster virus infection without skin involvement.

BACKGROUND: Varicella Zoster virus (VZV) infection is potentially very serious in bone marrow transplant recipients, and may manifest as a disseminated visceral infection. This condition is generally accompanied by a vesicular rash. OBJECTIVES: We review here a case of fulminant fatal disseminated VZV infection, not accompanied by skin involvement, and the laboratory approaches currently available to diagnose this disease. STUDY DESIGN: Post mortem tissue samples were subjected to histopathological examination, and tested for herpesviruses by electron microscopy and PCR. RESULTS: Intranuclear inclusions were noted by histological examination in the lungs, liver, kidneys and bone marrow. Particles with a herpesvirus morphology were visualized in liver tissue. VZV DNA was detected in liver and bone marrow by PCR followed by sequencing of the amplicons. Viremia was documented by retrospective testing of the serum by PCR. CONCLUSIONS: A disseminated VZV infection which proved rapidly fatal was demonstrated in a case without skin manifestations. This rare presentation of VZV infection is potentially underdiagnosed. Testing for VZV viremia by PCR can at the very least suggest the diagnosis although whether plasma-associated viremia is truly pathognomonic of visceral disseminated infection remains to be established.     

Transvaginal sonographic assessment of cervical length changes during triplet gestation

The current study aimed to evaluate the contribution of transvaginal sonography (TVS) for monitoring cervical changes during the second half of triplet gestation. Forty-five pregnant women with triplets pregnancies were prospectively scanned by TVS from approximately 26 weeks gestation and were longitudinally followed-up until delivery. Based on a receiver-operating curve it was found that a cervical length of 25 mm is the most accurate parameter (94% sensitivity and 45% specificity) for predicting premature delivery < or =33 gestational weeks. Thus, a single cervical length measurement of < or =25 mm at 26 weeks gestation correlated well with premature delivery at < or =33 weeks (chi(2); P = 0.002). Using the linear regression model, a mathematical equation [(Week of delivery = 27.4 + 1.6 x cervical length; R(2) = 0.46; P = 0.01)] for predicting the gestational age of delivery (dependent variable) was determined based on mid-gestation cervical measurements (predictors). In parturient women with triplet gestation, TVS assessment of the uterine cervix offers insight into the cervical status and provides valuable information for prenatal care. This includes both monitoring the cervical changes throughout third trimester as well as predicting the likelihood of premature delivery.     

4E-binding protein phosphorylation and eukaryotic initiation factor-4E release are required for airway smooth muscle hypertrophy

The molecular mechanisms of airway smooth muscle hypertrophy, a feature of severe asthma, are poorly understood. We previously established a conditionally immortalized human bronchial smooth muscle cell line with a temperature-sensitive SV40 large T antigen. Temperature shift and loss of large T cause G1-phase cell cycle arrest that is accompanied by increased airway smooth muscle cell size. In the present study, we hypothesized that phosphorylation of eukaryotic initiation factor-4E (eIF4E)-binding protein (4E-BP), which subsequently releases eIF4E and initiates cap-dependent mRNA translation, was required for airway smooth muscle hypertrophy. Treatment of cells with chemical inhibitors of PI 3-kinase and mammalian target of rapamycin blocked protein synthesis and cell growth while decreasing the phosphorylation of 4E-BP and increasing the binding of 4E-BP to eIF4E, consistent with the notion that 4E-BP1 phosphorylation and eIF4E function are required for hypertrophy. To test this directly, we infected cells with a retrovirus encoding a phosphorylation site mutant of 4E-BP1 (AA-4E-BP-1) that dominantly inhibits eIF4E. Upon temperature shift, cells infected with AA-4E-BP-1, but not empty vector, failed to undergo hypertrophic growth. We conclude that phosphorylation of 4E-BP, eIF4E release, and cap-dependent protein synthesis are required for hypertrophy of human airway smooth muscle cells.     

Angiogenic and lymphangiogenic microvessel density in breast carcinoma: correlation with clinicopathologic parameters and VEGF-family gene expression.

Angiogenesis and lymphangiogenesis are essential for breast cancer progression and are regulated by vascular endothelial growth factors (VEGF). To determine clinical and molecular correlates of these processes, we measured blood and lymphatic vascular microvessel density in 29 invasive carcinomas (22 ductal, six lobular, one papillary), using the vascular marker CD31 and the novel lymphatic marker D2-40. Microvessel density was assessed microscopically and by image cytometry, and was compared with tumor histology, grade, stage, lymph node metastasis, hormone receptors, HER2/neu status, and expression of VEGF, VEGF-C and VEGF-D by immunohistochemistry or quantitative RT-PCR. Strong correlation was observed between visual and image cytometric microvessel density using D2-40 but not CD31 (P=0.016 and 0.1521, respectively). Image cytometric CD31 microvessel density correlated with tumor size, grade, stage and lymph node metastasis (P=0.0001, 0.0107, 0.0035 and 0.0395, respectively). D2-40 microvessel density correlated with tumor stage (P=0.0123 by image cytometry) and lymph node metastasis (P=0.0558 by microscopy). Immunohistochemical VEGF signal in peritumoral blood vessels correlated with image cytometric CD31 and D2-40 microvessel density (P=0.022 and 0.0012, respectively), consistent with the role of VEGF in blood and lymphatic vascular growth. Intratumoral VEGF-C and VEGF-D expression by quantitative RT-PCR correlated with D2-40 (P=0.0291 by image cytometry) but not with CD31 microvessel density, which could suggest a selective role of VEGF-C and VEGF-D in lymphangiogenesis. CD31 and D2-40 microvessel density correlated significantly with several prognostic factors, including lymph node metastasis. Thus, measurements of angiogenesis and lymphangiogenesis may have utility for breast cancer pathology, particularly for estimation of metastatic risk.     

Multilocular renal cyst. A hamartoma with potential for neoplastic transformation?

Multilocular renal cyst is a pathologically distinctive lesion of uncertain pathogenesis that usually presents as a large abdominal mass in male infants or middle-aged women. We performed detailed light and electron microscopic studies of three nephrectomy specimens that contained multilocular cysts and attempted to correlate their morphology with various theories of pathogenesis. In one of our cases, a microscopic focus of clear cells that resembled renal cell adenocarcinoma was observed in the wall of one locule. Although such a finding has been reported previously, the lack of well-documented cases with metastases suggests that the prognosis for patients with this lesion remains uniformly excellent.     

Polyamines in Trichomonas vaginalis

Trichomonas vaginalis was grown in a modified Bushby's medium and putrescine, spermidine and spermine levels were determined in extracts from 24- and 48-h cultures and also in the culture media. All three polyamines were present in T. vaginalis extracts; the putrescine level and putrescine/spermidine ratio were much higher than those reported for other protozoa or for mammalian tissues. There were no significant differences between 24-h and 48-h amine levels per mg protein in these extracts, but amine levels per cell were higher at 24 than at 48 h. The spent culture media had a much higher putrescine content than corresponding uninoculated media and it was concluded that T. vaginalis secreted putrescine into the culture medium.     

Reflex inhibition of the quadriceps after meniscectomy: lack of association with pain

We have examined the severity and duration of reflex inhibition of quadriceps activation after arthrotomy and meniscectomy, its relationship with pain, and the effect of local anaesthesia on this relationship. Fourteen men, on completion of medial meniscectomy by arthrotomy, received either 10 ml (B10 group) or 15 ml (B15 group) of 0.5% bupivacaine hydrochloride ('Marcaine Plain') into the knee, or no injection (control group). Reflex inhibition of quadriceps was measured as the percentage reduction, from the ipsilateral preoperative value, in the integrated surface electromyogram recorded during maximal voluntary isometric contractions with the knee in extension. Pain during each contraction was recorded on a linear analogue scale. Unoperated limbs showed no evidence of quadriceps inhibition. In the operated limbs, at 1-2 h post-operatively, controls had both severe inhibition (median = 62%) and severe pain on attempting a maximal quadriceps contraction. The B10 group had similar inhibition but less pain (P less than or equal to 0.005, Wilcoxon 2-sample, 1-tailed test). In the B15 group both inhibition (P less than or equal to 0.05) and pain (P less than or equal to 0.01) were less than in the controls. These effects of bupivacaine had been lost by 4-5 h post-operatively. At 3-4 days, inhibition was still severe (median = 75%) in all three groups of patients but pain was only mild or absent. At 10-15 days, median inhibition was still 35%, but there was little or no pain. We conclude that postmeniscectomy inhibition is not simply due to perceived pain but is due, at least in part, to stimuli from the knee.