Identification of allele-specific KIV-2 repeats and impact on Lp(a) measurements for cardiovascular disease risk.

The abundance of Lp(a) protein holds significant implications for the risk of cardiovascular disease (CVD), which is directly impacted by the copy number (CN) of KIV-2, a 5.5 kbp sub-region. KIV-2 is highly polymorphic in the population and accurate analysis is challenging. In this study, we present the DRAGEN KIV-2 CN caller, which utilizes short reads. Data across 166 WGS show that the caller has high accuracy, compared to optical mapping and can further phase ~50% of the samples. We compared KIV-2 CN numbers to 24 previously postulated KIV-2 relevant SNVs, revealing that many are ineffective predictors of KIV-2 copy number. Population studies, including USA-based cohorts, showed distinct KIV-2 CN, distributions for European-, African-, and Hispanic-American populations and further underscored the limitations of SNV predictors. We demonstrate that the CN estimates correlate significantly with the available Lp(a) protein levels and that phasing is highly important.

The prevalence of Schmorl's nodes in osteoporotic vs normal patients: a Middle Eastern population study.

The study was conducted to establish the association of Schmorl's nodes and osteoporosis in a Middle Eastern cohort. The prevalence of SN in this sample was 41.1%. It was most frequent in the lumbar spine typically solitary central lesions. Over 88% Schmorl's node cases were osteoporotic/osteopenic and only 11.6% normal. INTRODUCTION: This study aims to identify the prevalence of Schmorl's nodes (SNs) in a cohort of Omani nationals, and also to determine any relation between osteoporosis and Schmorl's nodes. METHODS: This retrospective observational study was conducted on Omani nationals. One thousand three hundred and forty-eight DEXA scan patients were included. Of these, 545 patients had complete X-rays and MRI scans that would help determine the SN status. The X-rays and sagittal, coronal, and axial T2-weighted MR images were used to identify the presence and exact location of the Schmorl nodes by one orthopedic trainee and confirmed by the senior author. The correlation of each parameter with the presence of SN was analyzed by the independent-samples T test and one-way ANOVA. RESULTS: The overall prevalence of SN in this population sample appeared to be 41.1%. Over 88% of the SN-positive cases were either osteopenic or frankly osteoporotic by the WHO definition. Vast majority of SNs (87.1%) occurred in the lumbar spine and were central in location and mostly solitary. Statistical analysis of the data revealed significant correlation between osteopenia or osteoporosis and the presence of SNs. CONCLUSIONS: The prevalence of SN in the sample of Omanis studied was 41.1% and was most frequently seen in older men in the lumbar spine. It is strongly associated with osteoporosis/osteopenia (88.4%) and frequently presents as solitary central lesions.

Comparison of primers for the detection of genogroup II noroviruses in India.

PURPOSE: Noroviruses (NoV) are increasingly recognized as an important cause for acute gastroenteritis, worldwide. Reverse transcription polymerase chain reaction (RT-PCR) and sequencing are the methods of choice for the detection of NoVs, but there is currently no consensus about the primers to be used in these assays. MATERIALS AND METHODS: In this study, five published primer sets were evaluated for the detection of genogroup II (GII) NoVs in India. The primers target different regions of the NoV genome. Three primer sets detect an NoV in a single round RT-PCR platform, while the remaining two primer sets are based on a nested RT-PCR platform. RESULT: A panel of 100 samples from previous studies on norovirus diarrhoea in children were tested by all five primer sets. Of them, 74 samples were identified as positive for NoV, by at least one primer set. Subsets of positive amplicons were sequenced to check for specificity. CONCLUSION: The most sensitive primer set was Girish 2002, which detected GII NoV by nested RT-PCR, and was modified from the previously published primers. This study demonstrates that higher detection can be obtained by either using multiple primer sets or using a sensitive nested RT-PCR assay. It also demonstrates the differences in primer sensitivity for detection of Genogroup II (GII) NoVs in India.

Astrogliosis in culture. IV. Effects of basic fibroblast growth factor.

Previous studies have shown that the mechanical wounding of 3-week-old cultured rat astrocytes results in cell proliferation and hypertrophy resembling astrocyte responses to a brain injury in vivo. We now report the effects of basic fibroblast growth factor (bFGF) and an anti-bFGF antibody on astrocyte morphology, proliferation, and migration following in vitro wounding of confluent secondary cultures. Addition of bFGF (20 ng/ml) to wounded cultures induced morphological changes characteristic of differentiation in wounded and nonwounded areas of the culture. Combined treatment with bFGF and an anti-bFGF antibody (100 micrograms/ml) prevented this effect. Astrocyte proliferation along the edges of a scratch wound was at maximum 24 hr after wounding in cells growing in Eagle's minimum essential medium (EMEM) containing 10% serum. Low serum concentration and treatment with dibutyryl cyclic adenosine monophosphate (dbc-AMP) reduced injury-associated astrocyte proliferation. Addition of bFGF to cultures in EMEM with serum increased astrocyte proliferation at 18 and 24 hr after wounding. This effect was reduced considerably by treatment of cultures with bFGF in combination with an anti-bFGF antibody. The combined treatment and the antibody alone reduced cell division to a level lower than in control cultures. Twenty-four hr following wounding, astrocytes along the edges of the wound exhibited extension of thick, flat processes into the wound area. At 3 and 5 days after wounding, a bodily migration of astrocytes into the wounded area was observed. Addition of bFGF significantly increased astrocyte migration 1 day after wounding, with maximum effect on day 3 and no subsequent increase on day 5. A combination of bFGF and anti-bFGF antibody as well as the antibody alone reduced astrocyte migration to a level lower than in controls. Immunohistochemical localization and isoform pattern of bFGF in astrocytes did not change with dbc-AMP treatment or wounding. We conclude that mechanically wounded confluent astrocytes respond to bFGF added to the culture medium by enhancing cell division, differentiation, and migration. In addition, the results of the antibody treatment also suggest a role for endogenous bFGF in astrocyte proliferation and migration elicited by wounding in vitro. These results support the notion that in vivo, both bFGF released by injury and endogenous bFGF synthesized by astrocytes, contribute to the cellular responses that lead to astrogliosis.

Intralesion injection of basic fibroblast growth factor alters glial reactivity to neural trauma.

A traumatic injury to the rat brain is known to induce astrocyte proliferation and hypertrophy leading to astrogliosis. In addition, trauma also induces microglial proliferation in the brain. Since basic fibroblast growth factor (bFGF) is believed to play a role in mediating glial responses to brain injury, we examined the effects of bFGF administration on astrocyte and microglial proliferation and astrocyte hypertrophy resulting from a traumatic injury to the rat brain. Intracerebroventricular infusion of bFGF for 2 days prior to and following injury failed to alter glial reactivity. In contrast, a single intralesion injection of bFGF immediately after injury increased total cell division 2 and 5 days later, with an exclusive effect on glial fibrillary acidic protein-negative glia which consisted mostly of cells of macrophage-microglial phenotype. In addition, bFGF also enhanced injury-induced astrocyte hypertrophy. These results support a role for bFGF in macrophage-microglia proliferation and astrocyte hypertrophy following brain injury. They also suggest that alteration of injury responses of nonneuronal cells of the brain may be a potential side effect of intracerebral administration of bFGF.

A rapid procedure for the isolation of phycobilisomes from cyanobacteria.

This paper describes a method for the rapid isolation of phycobilisomes using a cationic detergent, CTAB (cetyltrimethylammonium bromide). The method has distinct advantages over those currently in use in that (i) release of intact phycobilisomes from cells in the presence of CTAB occurs in 40 s (as compared to 40-60 min of incubation required with Triton X-100), thereby reducing the chances of proteolysis of the component phycobiliproteins; and (ii) these phycobilisome preparations have reduced chlorophyll contamination in the initial stages. In addition this method also helps retain the structural and functional properties, as evidenced by spectroscopy and sodium dodecyl sulfate-polyacrylamide gel analysis.

Scapular reactivity to contact irritants: application-time and area variations.

Twenty-two diurnally active, fair-skinned, male volunteers were repeatedly patch tested with 10, 15 and 20% lapyrium chloride (LC) and 0.3 and 0.5% sodium lauryl sulfate (SLS) for 5 consecutive days. A series of 5 patches containing SLS and LC were placed vertically once daily for 6 h on different sites of the scapula: paravertebral, lateral and medial. The sites of application were randomized over subjects for patch testing once a day per area at either 08%, 16% or 00%. Each patch was left in place 6 h; scoring of each site, using a 13-point scale, was done 2, 10 and 18 h after removal. The cutaneous reactivity varied in a statistically significant manner according to the time and area of patch application. For each day except the first, LC scores exhibited highest and lowest responses at 16% and 00%, respectively. For SLS, a statistically significant application-time difference in reactivity was detected only for the readings of day 2 when scores were highest at 00% and lowest at 16%. Intraregional variations in scapular reactivity were also exhibited. Strongest reactions to LC occurred in the center of each scapula; weakest reactions were elicited on either the lateral or paravertebral sites. For SLS, statistically significant intraregional differences in reactivity were detected only for the second day of patching, with strongest responses near the spinal column and weakest near the axilla.

Adenosine 5'-triphosphate sulphurylase from Spirulina platensis.

ATP-sulphurylase from an unicellular blue-green alga, Spirulina platensis was localized in the soluble fractions of cell-free homogenate, and it was stable for over 3 weeks at -6 degrees C.