Early evolution of metazoan serine/threonine and tyrosine kinases: identification of selected kinases in marine sponges

The phylum Porifera (sponges) was the first to diverge from the common ancestor of the Metazoa. In this study, six cDNAs coding for protein- serine/threonine kinases (PS/TKs) are presented; they have been isolated from libraries obtained from the demosponges Geodia cydonium and Suberites domuncula and from the calcareous sponge Sycon raphanus. Sequence alignments of the catalytic domains revealed that two major families of PS/TK, the "conventional" (Ca(2+)-dependent) protein kinase C (PKC), the cPKC subfamily, as well as the "novel" (Ca(2+)- independent) PKC (nPKC), form two separate clusters. In each cluster, the sequence from S. raphanus diverges first. To approach the question about the origin of protein-tyrosine kinases (PTK), which are found only in Metazoa, we analyzed two additional PS/TKs which have been cloned from S. domuncula: the stress-responsive protein kinase (KRSvSD) and the protein-kinase-C-related kinase (PRKvSD). The construction of the phylogenetic tree, comprising the eight PS/TKs and the PTK cloned previously from G. cydonium, revealed that the PTK derived from the branch including the KRSvSD kinase. These data facilitate the first molecular approach to elucidate the origin of metazoan PTK within the PS/TK superfamily.      

Ambulatory Blood Pressure Monitoring in Individuals with HIV: A Systematic Review and Meta-Analysis

Introduction

Abnormal diurnal blood pressure (BP) rhythms may contribute to the high cardiovascular disease risk in HIV-positive (HIV⁺) individuals. To synthesize the current literature on ambulatory BP monitoring (ABPM) in HIV⁺ individuals, a systematic literature review and meta-analysis were performed.

Methods

Medical databases were searched through November 11, 2015 for studies that reported ABPM results in HIV⁺ individuals. Data were extracted by 2 reviewers and pooled differences between HIV⁺ and HIV-negative (HIV^-) individuals in clinic BP and ABPM measures were calculated using random-effects inverse variance weighted models.

Results

Of 597 abstracts reviewed, 8 studies with HIV⁺ cohorts met the inclusion criteria. The 420 HIV⁺ and 714 HIV^- individuals in 7 studies with HIV^- comparison groups were pooled for analyses. The pooled absolute nocturnal systolic and diastolic BP declines were 3.16% (95% confidence interval [CI]: 1.13%, 5.20%) and 2.92% (95% CI: 1.64%, 4.19%) less, respectively, in HIV⁺ versus HIV^- individuals. The pooled odds ratio for non-dipping systolic BP (nocturnal systolic BP decline <10%) in HIV⁺ versus HIV^- individuals was 2.72 (95% CI: 1.92, 3.85). Differences in mean clinic, 24-hour, daytime, or nighttime BP were not statistically significant. I² and heterogeneity chi-squared statistics indicated the presence of high heterogeneity for all outcomes except percent DBP dipping and non-dipping SBP pattern.

Conclusions

An abnormal diurnal BP pattern may be more common among HIV⁺ versus HIV^- individuals. However, results were heterogeneous for most BP measures, suggesting more research in this area is needed.     

Effect of I,25-dihydroxycholecalciferol in renal osteodystrophy.

A 23-year-old man with medullary cystic disease had been undergoing hemodialysis for 5 years and had become confined to a wheelchair because of renal osteodystrophy. He was treated with 125-dihydroxycholecalciferol, 2.0 mug (later 1.0 mug) three times a week, administered by way of the venous end of the dialysis machine. Within 1 month bone pain lessened and his ability to stand and walk improved. By 3 months he was walking short distances and by 5 months, long distances. Calcium balance was near zero before treatment and was strongly positive during treatment. Bone mineral content in the lower femur, measured by photon absorptiometry, increased at a rate of 32.2% per year. In contrast, 26 other patients on long-term hemodialysis had a mean loss of bone mineral content of 14.0% per year. Radiographs taken during treatment showed a decrease in subperiosteal bone resorption and healing of a pseudofracture. A significant decrease in the mean serum alkaline phosphatase value was noted during treatment, but no significant changes in mean serum calcium or phosphorus values were seen.     

STUDIES ON THE ORGANIZATION OF THE BRUSH BORDER IN INTESTINAL EPITHELIAL CELLS : II. Fine Structure of Fractions of Tris-Disrupted Hamster Brush Borders

Two of the fractions obtained by density gradient centrifugation of Tris-disrupted brush borders from hamster intestinal mucosa have been identified as the microvillus cores and their surrounding membranous coats, respectively. This identification has the following morphological basis. In shadowed preparations one fraction (cores) appears as rounded, compact rods, and the other fraction (coats) appears as flattened sheets. Both rods and sheets have dimensions appropriate to the identities assigned to them. In addition, negative staining shows that the rods are composed of aligned particles of roughly 60 A, consistent with the appearance of the core in tissue section, where 60-A fibrils are characteristic. The sheets are covered by non-aligned particles of approximately the same size. Sectioned preparations show that the core fraction contains predominantly fibrous material with some membranous contamination and that the coat fraction is apparently composed exclusively of elongated sacs with a unit membrane structure. Some details of the structure of the core are evident in cases where the compact rod appears to be loosened, revealing a doubled strand. The strand is approximately 350 A wide; the compact rod is roughly twice this width. With negative staining the strand shows a dense central region. The morphological identification presented here is consistent with the distribution of enzymic activity among the density gradient fractions described in the preceding paper.     

MICROTUBULES AND MICROFIBRILS IN MORPHOGENESIS OF THE SCALE CELLS OF EPHESTIA KÜHNIELLA

The development of scale cells in insects has been studied from the appearance of the first cytoplasmic projection which forms the scale rudiment. This rudiment contains numerous longitudinally oriented microtubules throughout. Immediately under its outer surface lie a series of adjacent but distinct bundles of longitudinally oriented circa 60-A fibrils with a circa 120-A center-to-center spacing. As the rudiment broadens, the microtubules become distributed near the surface. The rudiment finally becomes extremely broad and flattened. Fibril bundles are now widely separated and equally spaced. They still lie immediately below the cell surface. Then the cytoplasm protrudes midway between each fibril bundle to form longitudinal ridges and the major shape changes of the scale have been achieved. The final pattern can thus be related to the cytoplasmic organization of the rudiment. The main cytoplasmic elements which seem important in scale morphogenesis, on the basis of frequency, orientation and grouping, are 60-A fibrils and microtubules.     

Surveillance of vivax malaria vectors and civilian patients for malaria high-risk areas in northern Gyeonggi and Gangwon Provinces near the demilitarized zone,Republic of Korea, 2003–2006

After re-emergence of malaria in 1993, a continued increase in Plasmodium vivax cases was observed from 1993 to 2006 in northern Gyeonggi and Gangwon Provinces adjacent to the demilitarized zone separating North from South Korea. Annual parasite incidence per 1000 people ranged from 0.33 in 2004 to 0.89 in 2006. While malaria case rates declined (22.6%) in 2004, they increased 75.1% in 2005 and 51.7% in 2006 from the previous years. An initial incorrect diagnosis of 46.8% of malaria cases as common cold resulted in a mean delay of 1.3 days for the detection malarial parasites. Of the total cases, 10.2% from December to May were due to latent intrinsic incubation infections acquired the previous malaria season and the rest of the cases from June to November were either latent or short incubation infections. Overall, the peak anopheline population occurred from July to September, resulting in a similar peak in malaria cases. While malaria cases increased during 2005–2006, anopheline populations, based on trap indices, were not significantly different during 4 years of surveillance. To decrease the malaria patient infective period to mosquitoes, public health centers in Paju and Cheorwon in 2006 prescribed chloroquine + primaquine at days 0–3 after initial malaria diagnosis followed by an additional 11 days of primaquine (early primaquine treatment), rather than chloroquine on days 0–3 and primaquine on days 4–17 (delayed primaquine treatment). A reduction in the malaria parasite incidence during 2007 was recorded for the two locations offering the early primaquine treatment relative to other locations using the delayed primaquine treatment.     

Use of a stress-minimisation paradigm in high cell density fed-batch Escherichia coli fermentations to optimise recombinant protein production

Production of recombinant proteins is an industrially important technique in the biopharmaceutical sector. Many recombinant proteins are problematic to generate in a soluble form in bacteria as they readily form insoluble inclusion bodies. Recombinant protein solubility can be enhanced by minimising stress imposed on bacteria through decreasing growth temperature and the rate of recombinant protein production. In this study, we determined whether these stress-minimisation techniques can be successfully applied to industrially relevant high cell density Escherichia coli fermentations generating a recombinant protein prone to forming inclusion bodies, CheY–GFP. Flow cytometry was used as a routine technique to rapidly determine bacterial productivity and physiology at the single cell level, enabling determination of culture heterogeneity. We show that stress minimisation can be applied to high cell density fermentations (up to a dry cell weight of >70 g L^?1) using semi-defined media and glucose or glycerol as carbon sources, and using early or late induction of recombinant protein production, to produce high yields (up to 6 g L^?1) of aggregation-prone recombinant protein in a soluble form. These results clearly demonstrate that stress minimisation is a viable option for the optimisation of high cell density industrial fermentations for the production of high yields of difficult-to-produce recombinant proteins, and present a workflow for the application of stress-minimisation techniques in a variety of fermentation protocols.