Distinct patterns of expression of MHC class I and beta 2-microglobulin transcripts at early stages of mouse development.
The class I surface Ag recognized during tissue graft rejection are composed of a 40- to 45-kDa H chain and a 12-kDa L chain, beta 2-microglobulin (beta 2m). Regulation of MHC gene expression during early development is thought to play an essential role in maternal tolerance of the fetal allograft. Here we used in situ hybridization techniques to characterize the temporal and spatial pattern of expression of MHC class I mRNA in the developing mouse embryo. MHC class I transcripts were initially detected at day 9.5 postcoitus in the primary and secondary trophoblast giant cell populations. At this stage, none of the remaining fetal components of the developing placenta expresses MHC class I mRNA. By contrast, the outer zone of trophoblast giant cells is the only trophectoderm-derived tissue in the developing placenta that does not express detectable levels of beta 2m mRNA. These findings indicate that the onset of MHC class I and beta 2m gene expression in early postimplantation stage embryos is not coordinately regulated. In addition, we made use of pluripotent embryonic stem (ES) cells to study developmental regulation of MHC class I and beta 2m gene expression in vitro. ES cells can be induced to form endoderm-like cells by the technique of suspension culture. In contrast to retinoic acid-treated F9 cells, the endoderm derived from ES cells expressed high levels of beta 2m mRNA, but no detectable MHC class I transcripts. These findings are consistent with results of our in situ hybridization experiments showing high levels of expression of beta 2m mRNA, and extremely weak expression of MHC class I transcripts in derivatives of the visceral endoderm in vivo.


Telomere length and salivary cortisol stress reactivity in very preterm infants.
During the Neonatal Intensive Care Unit (NICU) stay, very preterm (VPT) infants are exposed to life-saving yet pain-inducing skin-breaking procedures (i.e., NICU pain-related stress) which contribute to the programming of hypo-responsive HPA axis development during the first months of life. Unfortunately, to date the mechanisms linking NICU pain-related stress and altered HPA axis regulation are only limitedly known. Telomere length (TL) regulation is an epigenetic mechanism previously shown to be affected by early stress exposures and capable of associating with HPA axis reactivity in children. In VPT infants, NICU pain-related stress was found to associate with decreased TL from birth to discharge, but there is no evidence for the association between TL and HPA axis in these infants. In this study, we prospectively examined the relationship between NICU pain-related stress and HPA axis reactivity to an age-appropriate socio-emotional condition (i.e., the Still-Face Procedure, SFP) in healthy VPT infants at 3-month corrected age. NICU pain-related stress was computed as the ratio between the number of skin-breaking procedures and length of NICU stay. A differential score (i.e., ∆TL) was obtained subtracting TL at birth from TL at discharge. A normalized (log10) cortisol reactivity index (CRI) was obtained by averaging post-stress (20 min after SFP) salivary cortisol sample on baseline value. A regression model controlling for neonatal and socio-demographic confounders showed that ∆TL was the only significant predictor of CRI. Although preliminary, these findings contribute to our knowledge of the mechanisms linking early exposures to adversity and later in life regulation of the HPA axis in VPT infants.


U.S. POSTDOCS: Report Urges Better Treatment, Status.
For years, U.S. postdocs have been complaining about paltry salaries, lack of benefits, and lowly status. This week, a committee of the National Academies of Sciences and Engineering and the Institute of Medicine validated many of their complaints and lent its considerable weight to efforts to provide greater institutional support for postdocs. At the same time, however, the panel sidestepped two burning issues by explicitly declining to recommend a boost in postdoc salaries or take a position on whether to curtail the size of the postdoc workforce, which has more than doubled in the past 20 years to an estimated 52,000.


Cerebrovascular lesions in stroke-prone spontaneously hypertensive rats.
The cerebrovascular lesions of severe chronic hypertension were studied by light microscopy in perfusion-fixed, subserially sectioned brains from stroke-prone spontaneously hypertensive rats (SHRSP). The leakage and spread of plasma proteins were visualized by immunohistochemical detection of extravasated fibrinogen and by using an exogenous marker (Evans blue injected i.v.) for blood-brain barrier (BBB) dysfunction. In most SHRSP the hypertension did not lead to major BBB lesions in spite of a mean arterial pressure around 200 mm Hg at 6-9 months of age. Multifocal BBB damage occurred in a minor group of SHRSP, particularly within the cortex and the deep gray matter. A close spatial correlation was found between the leakage-spread of plasma constituents and the neuropathologic alterations. Fibrinoid degeneration of penetrating arterioles was found within the leakage sites. The surrounding gray matter showed petechial hemorrhages and abundant proteinaceous exudates rich in antifibrinogen-positive material. The current leakage of Evans blue and wide spread of fibrinoid substances suggested long-lasting damage to the BBB. Most neurons within the edematous gray matter had well preserved nuclei surrounded by a rim of cytoplasm with ill-defined outline as if vacuolation or lysis of the peripheral cytoplasm had occurred. The sponginess of the tissue progressed in severe cases to formation of necrotic cysts. Condensed acidophilic neurons were seen in the border zone between the edematous and more compact gray matter. The appearance and distribution of the gray matter lesions deviated in many respects from those commonly seen in regional ischemic infarcts. The fibrin thrombi found close to the cysts might be regarded as secondary events. The extensive spread of antifibrinogen-positive material within the white matter seemed to originate mainly from the chronic leakage sites in the gray matter. Increased number of large astrocytes were seen within the leakage sites and along the spreading pathways for the edema constituents. The white matter showed a rarefied texture with widely dispersed nerve fiber tracts, volume expansion, and occasional cyst formation. The results indicate a crucial pathophysiologic role for the egress, spread, and accumulation of vasogenic edema in the development of the cerebrovascular lesions in SHRSP.


Motor disabilities and effects of thyrotropin-releasing hormone on ataxia of spinocerebellar degeneration.
Fine aiming task was studied quantitatively in 20 patients with spinocerebellar degeneration (SCD) and 8 normal control subjects, utilizing a manual control system with random-step input signal. The transfer function of the system was an integral function and ability to perform a 'differentiating action' compensating for this integral function was analyzed. This 'differentiating action' was defective in all patients with SCD and was normal in all control subjects. A display of the dynamic characteristics of subjects on the plane defined by principal component analysis of the parameters showed definite delineation of the characteristics of patients from those of the controls. The intramuscular injection of thyrotropin-releasing hormone (TRH) resulted in an increase in amplitude of manipulation, a shortening of reaction time and consequently an improvement in motor disabilities in 65% of 20 patients with SCD.


Current concepts and diagnostic evaluation of autoimmune disease.
This article discusses autoimmune reactions and the numerous mechanisms by which an autoimmune response may be initiated, including genetic factors, T-cell bypass mechanisms, and idiotypes. Human autoimmune diseases are classified into three main groups, ie, organ-specific, non-organ specific, and disorders with non-organ-specific autoantibodies with lesions restricted to one or a few organs, that are examined in detail. General laboratory tests and interpretation of results in relation to state or treatment of the particular disease, age and sex of the patient, and the sensitivity of the test system used are reviewed.


[Long-term results of traditional periodontal therapy].
To evaluate the long-term results of traditional therapy consisting of initial preparation followed by mucogingival surgery including bone recontouring and regular maintenance treatments. 125 patients between 25 and 70 with a generalised periodontitis of the type adult and rapidly progressing periodontitis were selected. These patients were treated according to a fixed protocol by a periodontist and a dental hygienist. The Practice for Periodontology in Groningen. Patients were treated in a practice limited to periodontology. Before and 5 years after treatment the following data were assessed: plaque- and bleeding-score, probing pocket-depth, mobility and furcation involvement. Furthermore the reason for extraction was established. In general a great reduction in plaque- and bleeding-score and probing pocket-depth was much better in case of good oral hygiene. Within 5 years after treatment only 1.1% of the teeth were extracted. Furcations with through and through involvement in combination with deep residual pockets were mostly the reason for this. It is concluded that with traditional periodontal therapy following a strict protocol including regular maintenance treatments excellent treatment results can be achieved.


PTH inhibition of bicarbonate transport by proximal convoluted tubules.
These studies examined the effect of parathyroid hormone (PTH), dibutyryl cyclic AMP DBcAMP, and 8-bromo-cyclic AMP BrcAMP on HCO3- transport by rabbit superficial proximal convoluted tubules perfused in vitro. Bicarbonate was estimated as total CO2 measured microcalorimetrically. At slow perfusion rates with 25 mM HCO3- in the perfusate and bath, PTH (0.1 U/ml in the bath) caused the total CO2 in tubular fluid to rise from 10.2 to 19.9 mM. The hormone had no effect on the total CO2 concentration in tubules perfused with HCO3(-)-free perfusates. With HCO3(-) in the perfusate and bath, PTH reduced the rates of fluid and total CO2 absorption to 57 and 48% of control values, respectively. PTH had no effect on the rates of fluid absorption and total CO2 secretion when HCO3(-)-free perfusates were used. The effects of DBcAMP and BrcAMP (10(-7) M in the bath) were similar to those of PTH. 5'-AMP (10(-6) M in the bath) did not alter the total CO2 concentration of tubular fluid when the tubules were perfused at slow rates with HCO3- in the perfusate and bath. Ouabain (10(-5) M in the bath) caused the total CO2 concentration in tubules perfused at slow rates with HCO3--free perfusates to rise from 8.9 to 12.7 mM. PTH caused no further change in the total CO2 concentration in the presence of ouabain.


Flat topped beams and their characteristics in turbulent media.
The source and receiver plane characteristics of flat topped (FT) beam propagating in turbulent atmosphere are investigated. To this end, source size, beam power and M(2) factor of source plane FT beam are derived. For a turbulent propagation medium, via Huygens Fresnel diffraction integral, the receiver plane intensity is found. Power captured within an area on the receiver plane is calculated. Kurtosis parameter and beam size variation along the propagation axis are formulated. Graphical outputs are provided displaying the variations of the derived source and receiver plane parameters against the order of flatness and propagation length. Analogous to free space behaviour, when propagating in turbulence, the FT beam first will form a circular ring in the center. As the propagation length increases, the circumference of this ring will become narrower, giving rise to a downward peak emerging from the center of the beam, eventually turning the intensity profile into a pure Gaussian shape.


Effect of soy proteins and isoflavones on lipid metabolism and involved gene expression.
Clinical trials and animal studies showed that ingestion of soy proteins improves blood lipid profiles including lowering triglyceride, total and LDL cholesterol levels and increasing HDL cholesterol content. However, the effective components in the soy and the mechanisms involved in the hypolipidemic actions are not fully understood. Increasing evidence from animal studies have suggested that soy components may regulate lipid metabolism by modulating the activities of key transcription factors and thereby changing the downstream gene expression involved in lipogenesis or lipolysis. It has been shown that intake of soy proteins alters the expression of genes for sterol regulatory element binding protein, peroxisomal proliferator activated receptor, and liver X receptor. Dietary soy proteins suppress the DNA binding activities of hepatic nuclear receptors for thyroid hormones and retinoic acid, and alter the activities of key enzymes including cholesterol 7alpha hydroxylase and ATPase/ATP synthase through post-translational protein modifications. This paper reviews the current understanding of the cellular and molecular events by which soy components affect lipid levels, especially focusing on modulation of transcription factors and regulation of gene expression involved in lipid metabolism by soy proteins and associated isoflavones.


A2A Adenosine Receptor Agonists and their Potential Therapeutic Applications. An Update.
Adenosine is an endogenous purine nucleoside, which mediates a variety of important biological processes and diseases, such as vasodilation, inflammation, cancer, wound healing, ischemia reperfusion injury, Parkinson disease, infectious diseases, and other CNS disorders. Particularly important are the A2A receptors that have been expressed in the lung, liver, heart, cardiovascular tissues, leukocytes, neutrophils, and endothelial cells. This review provides an update of the latest A2A receptor agonists developed in the period 2005-2017, their selectivity regarding other adenosine receptors and their potential therapeutic applications. I have conducted an extensive search from the most common bibliographic databases for critically review the most recent works on the A2A receptor agonists and their therapeutic applications in inflammation, asthma and chronic obstructive pulmonary disease, myocardial perfusion imaging, sepsis, rheumatoid arthritis, and wound healing, among others. In the last decade, a great deal of effort has been devoted to develop adenosine receptor agonists and antagonists for treatment of a number of diseases. Thus, for A2A receptor agonists more than 130 papers and reviews have been found, many of them highlighting the usefulness of these compounds in the field. Although so far many of the A2A receptor agonists have failed in clinical trials due to their side effects, some of them have been approved for protection against cardiac ischemia-reperfusion injury and anemia. The recently reported crystal structure of the human A2A receptor in complex with the agonist UK-432097 is a fundamental keystone for the development of new and selective A2A ligands with new therapeutic applications.


The impact of left temporal lobe seizure disorder on learning disorders: a case study.
This case study describes the relationship between left temporal lobe epilepsy (TLE) and learning disabilities in a 26-year-old male college student. The client developed seizures following an episode of mycoplasma encephalitis at the age of 7. The client underwent a left temporal lobectomy involving resection of the left mesial temporal lobe, amygdala, hippocampus, and part of the brain stem 6 years prior to the current evaluation, in an attempt to address the frequency of the seizures. The surgery was extensive, including neocortical resection extending posterior to the vein of Labbe along the inferior temporal gyrus. The lobectomy reportedly successfully eliminated the seizures and the need for anti-seizure medications, but no neurological or neuropsychological follow-up occurred until 2009 when he was referred by his academic program for an evaluation of learning disabilities. Results of the neuropsychological evaluation indicated significant expressive language functioning deficits, with generally better-preserved receptive language. However, compared to a pre-surgical neuropsychological evaluation there was evidence for subtle to mild improvement in several aspects of cognitive functioning, likely due to seizure elimination and discontinuation of the anti-seizure medication. Nonetheless, his deficits resulted in significant functional impact on his academic abilities, thus implications for academic intervention were discussed.


Combination of Transforaminal and Interlaminar Percutaneous Endoscopic Lumbar Diskectomy for Extensive Down-migrated Disk Herniation.
The technique and instrumentation of percutaneous endoscopic lumbar diskectomy (PELD) have significantly improved. Thus its indications have been gradually expanding. Down-migrated disk, in particular, is regarded inaccessible by rigid instrumentation due to poor visualization and limited accessibility. We introduce a combination of transforaminal and interlaminar PELD for extensive down-migrated disk herniation at the L4-L5 level. In the first case, a 48-year-old man had left L5 radicular pain. Magnetic resonance imaging (MRI) showed that L4-L5 disk herniation extended to the L5 lower end-plate level. In the second case, a 39-year-old man presented with right L5 and S1 radiculopathy. MRI showed right huge extensive down-migrated disk herniation from the L4-L5 disk space to the S1 pedicle level. Back pain and leg pain were measured using the visual analog scale (VAS). In the first case, transforaminal PELD with foraminoplasty removed the disk fragment from the L4-L5 disk space to the mid-L5 pedicle level. Interlaminar PELD removed the remaining disk below the mid-L5 pedicle. In the second case, first, contralateral (left) transforaminal PELD at L4-L5 removed disk fragments located at the subannular and subligamentous area as well as the upper part of the down-migrated disk herniation. Second, interlaminar PELD via an ipsilateral (right) L5-S1 interlaminar space removed the remains of the extensively down-migrated disk herniation. In the first case, the VAS scores for back and leg pain were improved from 6 and 8 to 2 and 1, respectively. Postoperative MRI showed complete removal of the disk fragment. In the second case, the VAS scores for back and leg pain improved from 7 and 9 to 3 and 1, respectively, after PELD. Postoperative MRI showed complete removal of a huge disk herniation. Both patients were discharged the day after PELD. Follow-up examinations showed no recurrence. It is difficult to remove entire disk fragments using only a transforaminal or interlaminar approach for extensive down-migrated disk herniation. Therefore a combination of transforaminal and interlaminar PELD may be effective for extensive down-migrated disk herniation at L4-L5.


Anti-renal cell carcinoma chimeric antibody G250: cytokine enhancement of in vitro antibody-dependent cellular cytotoxicity.
The chimeric monoclonal antibody cG250 targets the G250 antigen, a transmembrane protein which is expressed on renal carcinoma cells and is identical to the MN/CAIX antigen. In vitro studies have previously demonstrated that cG250 induces antibody-dependent cellular cytotoxicity (ADCC) of G250-positive targets. In order to investigate the upregulation of ADCC mediated by cG250, ADCC was examined using effector cells cultured in the presence or absence of the cytokines interferon-gamma (IFN-gamma), interferon-alpha isoforms IFN-alpha (2a) and IFN-alpha (2b) and interleukin-2 (IL-2), and the time course of effects over a 7-day period was determined. Renal cell carcinoma lines expressing high (SK-RC-52) and low (SK-RC-09) G250 antigen levels were used as target cells, and freshly isolated peripheral blood mononuclear cells (PBMC) from a healthy donor were used as the effector cells. PBMC were incubated with the respective cytokine at a range of concentrations or with a media alone control for a period of 7 days. The ADCC activity mediated by cG250 or control isotype matched huA33 with the different PBMC treatment groups was assessed in triplicate daily. Corresponding lymphokine activated killing (LAK) activity was measured concurrently for each treatment group. Chimeric G250 specifically recognised G250 antigen on high and low expressing cell lines SK-RC-52 and SK-RC-09, and mediated specific in vitro ADCC of both lines. In the absence of cytokine stimulation, the specific ADCC of cG250 declined rapidly within three days. IL-2 strongly enhanced and maintained cG250-mediated ADCC activity and K562 cytotoxicity when applied to PBMC in culture for seven days. IFN-gamma also enhanced the ADCC of cG250 throughout the study period, but was not as effective as the IL-2 treatment, and the SK-RC-09 line displayed lower specific cytotoxicity than the SK-RC-52 cell line. In contrast, IFN-alpha 2a and 2b increased cG250-mediated ADCC and K562 cytotoxicity for only three days of the study period. The potent and sustained immune effector activity observed with cG250 and cytokines in this in vitro study suggests that the combination immunotherapy of cG250 with cytokines such as IL-2 shows promise in the treatment of renal cell carcinoma (RCC).


The gene expression profiles of canine mammary cancer cells grown with carcinoma-associated fibroblasts (CAFs) as a co-culture in vitro.
It is supposed that fibroblasts present in tumour microenvironment increase cancer invasiveness and its ability to metastasize but the mechanisms have not been clearly defined yet. Thus, the current study was designed to assess changes in gene expression in five various cancer cell lines grown as a co-culture with the carcinoma-associated fibroblasts (CAFs) in vitro. A carcinoma-associated fibroblast cell line was isolated from a canine mammary cancer. Then, a co-culture of cancer cells with the CAFs was established and maintained for 72 hrs. Having sorted the cells, a global gene expression in cancer cells using DNA microarrays was examined. The analysis revealed an up-regulation of 100 genes and a down-regulation of 106 genes in the cancer cells grown as a co-culture with the CAFs in comparison to control conditions. The PANTHER binomial statistics tool was applied to determine statistically over-manifested pathways (p < 0.05). Bulk of the up-regulated genes are involved in the adhesion, the angiogenesis, the epithelial-mesenchymal transition (EMT) and generally take part in the developmental processes. These results were further confirmed using real-time qPCR. Moreover, a wound-healing assay and growth characteristics on Matrigel matrix showed that CAFs increase cancer cell migration and matrix invasion. The results of the current study showed that the co-culturing of cancer cells and the CAFs caused significant changes to the cancer gene expression. The presence of the CAFs in a microenvironment of cancer cells promotes adhesion, angiogenesis and EMT.


Vitamin K-dependent carboxylation of coagulation factors: insights from a cell-based functional study.
Vitamin K-dependent carboxylation is a post-translational modification essential for the biological function of coagulation factors. Defects in carboxylation are mainly associated with bleeding disorders. With the discovery of new vitamin K-dependent proteins, the importance of carboxylation now encompasses vascular calcification, bone metabolism, and other important physiological processes. Our current knowledge of carboxylation, however, comes mainly from in vitro studies carried out under artificial conditions, which have a limited usefulness in understanding the carboxylation of vitamin K-dependent proteins in native conditions. Using a recently established mammalian cell-based assay, we studied the carboxylation of coagulation factors in a cellular environment. Our results show that the coagulation factor's propeptide controls substrate binding and product releasing during carboxylation, and the propeptide of factor IX appears to have the optimal affinity for efficient carboxylation. Additionally, non-conserved residues in the propeptide play an important role in carboxylation. A cell-based functional study of naturally occurring mutations in the propeptide successfully interpreted the clinical phenotype of warfarin's hypersensitivity during anticoagulation therapy in patients with these mutations. Unlike results obtained from in vitro studies, results from our cell-based study indicate that although the propeptide of osteocalcin cannot direct the carboxylation of the coagulation factor, it is required for the efficient carboxylation of osteocalcin. This suggests that the coagulation factors may have a different mechanism of carboxylation from osteocalcin. Together, results from this study provide insight into efficiently controlling one physiological process, such as coagulation without affecting the other, like bone metabolism.


The effect of UV-B irradiation on secondary epidermal infection of mice with herpes simplex virus type 1.
Previous studies have indicated that suberythemal ultraviolet B (UV-B) irradiation of C3H mice before primary infection with herpes simplex virus (HSV) type 1 does not result in increased morbidity or mortality, but a suppressed delayed type hypersensitivity (DH) to the virus can be demonstrated. Any effect of UV radiation on pathogenesis during secondary epidermal HSV infection has not been previously examined. Mice were immunized by subcutaneous injection of inactivated HSV and, 5 days later, one group was UV-B-irradiated. The next day all mice were challenged epidermally with HSV. Most of the mice (92%) in the irradiated group developed severe lesions, whilst 59% of the non-irradiated group had mild lesions and 30% no lesions. Infectious virus was not isolated from the adrenal glands after challenge in either group. In addition, the DH to the virus was not affected by the UV exposure. The numbers of lymphocytes and dendritic cells in the lymph nodes draining the site of epidermal infection were increased in the UV group compared with the non-irradiated group. Following challenge, the percentage of CD4+ and CD8+ lymphocytes in lymph nodes was unaltered but the MHC class II expression on dendritic cells in these lymph nodes was reduced by UV exposure. The lymphoproliferative response in vitro of lymph node cells revealed a suppressed response to HSV and to the mitogen concanavalin A in the irradiated group. Thus, UV irradiation prior to epidermal secondary infection with HSV led to more severe infections due, perhaps, to a modulation in local antigen presentation.


Anti-inflammatory and anti-arthritic effects of Yucca schidigera: a review.
Yucca schidigera is a medicinal plant native to Mexico. According to folk medicine, yucca extracts have anti-arthritic and anti-inflammatory effects. The plant contains several physiologically active phytochemicals. It is a rich source of steroidal saponins, and is used commercially as a saponin source. Saponins have diverse biological effects, including anti-protozoal activity. It has been postulated that saponins may have anti-arthritic properties by suppressing intestinal protozoa which may have a role in joint inflammation. Yucca is also a rich source of polyphenolics, including resveratrol and a number of other stilbenes (yuccaols A, B, C, D and E). These phenolics have anti-inflammatory activity. They are inhibitors of the nuclear transcription factor NFkappaB. NFkB stimulates synthesis of inducible nitric oxide synthase (iNOS), which causes formation of the inflammatory agent nitric oxide. Yucca phenolics are also anti-oxidants and free-radical scavengers, which may aid in suppressing reactive oxygen species that stimulate inflammatory responses. Based on these findings, further studies on the anti-arthritic effects of Yucca schidigera are warranted.


The estimand framework had implications in time to patient-reported outcomes deterioration analyses in cancer clinical trials.
To apply the estimand framework in time to deterioration (TTD) analysis of patient-reported outcomes (PROs), and identify the appropriate statistical methods to deal with intercurrent event (IEs) such as death. Data from phase II randomized trial were used. We estimated TTD using European Organization for Research and Treatment of Cancer Quality of Life Questionnaire-C30 questionnaire with death as the IE, by applying Kaplan-Meier (K.M.) estimator and Cox proportional hazards (PH) model. The Fine-Gray approach was explored, accounting for death as a competing risk. The estimands targeted by the aforementioned methods were defined. We analyzed the data of 64 patients with available questionnaires at baseline. The most notable differences in TTD estimates were observed for deterioration in physical functioning: the hazard ratios were 0.44 [95% CI 0.22-0.90] and 0.62 [95% CI 0.36-1.07] by either ignoring death (31 events) or considering it as deterioration (58 events), respectively (Cox-PH model). When considering death as a competing event (Fine-Gray model), the sub-HRs was 0.51 [95% CI 0.26-1.01]. Depending on the proportion and distribution of deaths occurring before deterioration between arms, the Fine-Gray competing risks model should be considered rather than KM estimator and Cox PH model to reflect the patient's experience of the disease and treatment burden.


Orientation and alignment control of microphase-separated PS-b-PDMS substrate patterns via polymer brush chemistry.
Block copolymer (BCP) microphase separation at substrate surfaces might enable the generation of substrate features in a scalable, bottom-up fashion, provided that the pattern structure, orientation, and alignment can be strictly controlled. The PS-b-PDMS (polystyrene-b-polydimethylsiloxane) system is attractive because it can form small features and the two blocks can be readily differentiated during pattern transfer. However, PS-b-PDMS offers a considerable challenge, because of the chemical differences in the blocks, which leads to poor surface wetting, poor pattern orientation control, and structural instabilities. These challenges are considerably greater when line patterns must be created, and this is the focus of the current work. Here, we report controlled pattern formation in cylinder-forming PS-b-PDMS by anchoring different types of hydroxyl-terminated homopolymer and random copolymer brushes on planar and topographically patterned silicon substrates for the fabrication of nanoscale templates. It is demonstrated that non-PDMS-OH-containing brushes may be used, which offers an advantage for substrate feature formation. To demonstrate the three-dimensional (3-D) film structure and show the potential of this system toward applications such as structure generation, the PDMS patterns were transferred to the underlying substrate to fabricate nanoscale features with a feature size of ~14 nm.


The SSR-based molecular profile of 1005 grapevine (Vitis vinifera L.) accessions uncovers new synonymy and parentages, and reveals a large admixture amongst varieties of different geographic origin.
A collection of 1005 grapevine accessions was genotyped at 34 microsatellite loci (SSR) with the aim of analysing genetic diversity and exploring parentages. The comparison of molecular profiles revealed 200 groups of synonymy. The removal of perfect synonyms reduced the database to 745 unique genotypes, on which population genetic parameters were calculated. The analysis of kinship uncovered 74 complete pedigrees, with both parents identified. Many of these parentages were not previously known and are of considerable historical interest, e.g. Chenin blanc (Sauvignon × Traminer rot), Covè (Harslevelu selfed), Incrocio Manzoni 2-14 and 2-15 (Cabernet franc × Prosecco), Lagrein (Schiava gentile × Teroldego), Malvasia nera of Bolzano (Perera × Schiava gentile), Manzoni moscato (Raboso veronese × Moscato d'Amburgo), Moscato violetto (Moscato bianco × Duraguzza), Muscat of Alexandria (Muscat blanc à petit grain × Axina de tres bias) and others. Statistical robustness of unexpected pedigrees was reinforced with the analysis of an additional 7-30 SSRs. Grouping the accessions by profile resulted in a weak correlation with their geographical origin and/or current area of cultivation, revealing a large admixture of local varieties with those most widely cultivated, as a result of ancient commerce and population flow. The SSRs with tri- to penta-nucleotide repeats adopted for the present study showed a great capacity for discriminating amongst accessions, with probabilities of identity by chance as low as 1.45 × 10(-27) and 9.35 × 10(-12) for unrelated and full sib individuals, respectively. A database of allele frequencies and SSR profiles of 32 reference cultivars are provided.


Molecular Mechanism of Polycation-Induced Pore Formation in Biomembranes.
Polycations are an attractive class of macromolecules with promising applications as drug/gene carriers and biocides. The chemical structure and concentration of a polycation determine its interaction with cellular membranes and, hence, are crucial parameters for designing efficient nontoxic polycations. However, the interaction of polycations with biomembranes at the molecular level and the corresponding free-energy landscape is not well understood. In this work, we investigate the molecular mechanism of interaction between a strong polycation substituted with alkyl moieties and zwitterionic membranes via long-time-scale all-atom molecular dynamics simulations and free-energy calculations combined with Langmuir monolayer, atomic force microscopy, and calcein-release experimental measurements. We found that the membrane activity of the polycation and its ability to induce pores in the membranes can be attributed to the polycation-induced changes in the bilayer organization, such as reduced membrane thickness, increased disorder of the acyl chains, reduced packing, and electrostatic field gradients between membrane leaflets. These changes facilitate the penetration of water into the membrane and the formation of aqueous defects/pores. The calculated free-energy profiles indicate that the polycation lowers the nucleation barrier for pore opening and the free energy for pore formation in a concentration-dependent manner. Above the critical coverage of the membrane, the polycation nucleates spontaneous pores in zwitterionic membranes. Our work demonstrates the potential of combining enhanced sampling methods in MD simulations with experiments for a quantitative description of various events in the polycation-membrane interaction cycle, such as strong adsorption on the membrane due to hydrophobic and electrostatic interactions, and pore formation.


[Current and unresolved problems in the classification of cranio-cerebral injuries].
A careful study of the literature of the recent years and a special questionary presented in 1974 to 146 leading scientists and specialists of the country revealed important problems for discussion, such as contradictory approaches to the differentiation of the prevailing forms of lesions--commotion and contusion of the brain--by the degree of their severity, to distinguishing between brain contusion and compression. A prognosis of the tendencies and pathways to the creation of a common up-to-date classification of craniocerebral injuries is made.


Fieldwork experience, Part I: Impact on practice preference.
Occupational therapy students and student supervisors in 65 fieldwork centers nationwide responded to a survey designed a) to appraise the relative value of the fieldwork experience in the occupational therapist's professional development and b) to identify the elements perceived as being essential to a good fieldwork experience. Three stages of professional development were examined: the preprofessional, the academic, and the fieldwork experience. Results suggest that the fieldwork experience stage has the greatest impact on the development of a therapist's preference for a specific area of clinical practice. This article discusses the primary influences in the three stages that contribute to this preference formation.


[Quality of life: CVA patients take stock].
The approach of the concept 'quality of life' in stroke research is often multi-dimensional; and the dimensions to be studied, are chosen by the researcher. To determine the most important dimensions or aspects of quality of life--from the point of view of stroke patients--we interviewed 75 patients one year, and 83 patients two years after their stroke. These interviews were part of a longterm follow-up of a cohort of 221 stroke patients. Seventy-five percent of both the one- and two-year survivors experienced a deterioration of quality of life. Autonomy appeared to be the most important aspect of quality of life, being spontaneously mentioned by 55% of 75 patients. Compared with the stroke patients, an age- and sex-matched group of 61 controls rated their quality of life more highly. For them, autonomy appeared to be less important, being spontaneously mentioned by 25% of 61 controls. Furthermore, stroke patients attached more importance to social contacts and activities and general health than controls. We suggest to lend more weight in research and medical care, to patients' priorities in quality of life.


Dementia associated with dorsal midbrain lesion.
Although the dorsal midbrain has been implicated in cognitive processes in animals, its role in humans is unclear. We report the neuropsychological and postmortem neuropathological findings of a 52-yr-old university professor who developed a profound dementia in association with a focal dorsal midbrain lesion. The patient's disorder appeared to result from a tuberculous granuloma based on the clinical course and autopsy results. Neuropsychologically, he exhibited a generalized impairment across most of the cognitive domains assessed. His deficits were not explained by impaired arousal, specific sensory or motor defects, depression, or hydrocephalus. Although there are inherent limitations to a single-case investigation, our observations are consistent with animal studies that have demonstrated that focal dorsal midbrain lesions may result in cognitive impairment. We propose that the dorsal midbrain is involved in cognitive processing via modulation of thalamocortical networks.


Kinetic Deposition of Polar and Non-polar Lipids on Silicone Hydrogel Contact Lenses.
Purpose: This study investigated kinetic lipid uptake to four silicone hydrogel (SiHy) lenses over a period of four weeks, using an in-vitro radiolabel method. Methods: Four contemporary monthly replacement SiHy lenses (lotrafilcon B, senofilcon C, comfilcon A, samfilcon A) were incubated in three different solutions: 1) An artificial tear solution (ATS) containing 14C-labeled phosphatidylcholine (PC), 2) an ATS containing 14C-cholesteryl oleate (CO) and 3) an ATS containing four 14C-radiolabeled lipids (PC, phosphatidylethanolamine, CO, and cholesterol (total lipid)). After 16 hours, lipids were extracted twice from the lenses with chloroform:methanol and the radioactive counts determined the lipid quantities to simulate 1 day of wear. OPTI-FREE PureMoist (Alcon) was used to clean and disinfect the remaining lenses daily and the lipid quantities were further determined after 2 weeks and 4 weeks. Results: The amount of total lipid increased for all lenses over time (p < .01). After four weeks, total lipid accumulated was 20.26 ± 0.15 µg/lens for senofilcon C, which was significantly higher (p < .01) than all other lens materials (samfilcon A - 17.84 ± 0.21; comfilcon A - 16.65 ± 0.12; lotrafilcon B - 7.41 ± 0.56 µg/lens). CO was highest on lotrafilcon B (1.26 ± 0.13 µg/lens) and senofilcon C attracted the most PC (3.95 ± 0.12 µg/lens) compared to the other materials. Conclusion: The amount of both polar and non-polar lipid deposition on monthly replacement SiHy lenses increased over 4 weeks, with significant differences being seen between lens materials.


Standardized Definitions for Evaluation of Acute Decompensated Heart Failure Therapies: HF-ARC Expert Panel Paper.
Acute decompensated heart failure (ADHF) is one of the most common reasons for hospitalizations or urgent care and is associated with poor outcomes. Therapies shown to improve outcomes are limited, however, and innovation in pharmacologic and device-based therapeutics are therefore actively being sought. Standardizing definitions for ADHF and its trajectory is complex, limiting the generalizability and translation of clinical trials to effect clinical care and policy change. The Heart Failure Collaboratory is a multi-stakeholder organization comprising clinical investigators, clinicians, patients, government representatives (including U.S. Food and Drug Administration and National Institutes of Health participants), payors, and industry collaborators. The following expert consensus document is the product of the Heart Failure Collaboratory convening with the Academic Research Consortium, including members from academia, the U.S. Food and Drug Administration, and industry, for the purposes of proposing standardized definitions for ADHF and highlighting important endpoint considerations to inform the design and conduct of clinical trials for drugs and devices in this clinical arena.


Fractionated Radiation Therapy Stimulates Antitumor Immunity Mediated by Both Resident and Infiltrating Polyclonal T-cell Populations when Combined with PD-1 Blockade.
Purpose: Radiotherapy is a highly effective anticancer treatment forming part of the standard of care for the majority of patients, but local and distal disease recurrence remains a major cause of mortality. Radiotherapy is known to enhance tumor immunogenicity; however, the contribution and mechanisms of radiotherapy-induced immune responses are unknown.Experimental Design: The impact of low-dose fractionated radiotherapy (5 × 2 Gy) alone and in combination with αPD-1 mAb on the tumor microenvironment was evaluated by flow cytometry and next-generation sequencing of the T-cell receptor (TCR) repertoire. A dual-tumor model was used, with fractionated radiotherapy delivered to a single tumor site to enable evaluation of the local and systemic response to treatment and ability to induce abscopal responses outside the radiation field.Results: We show that fractionated radiotherapy leads to T-cell infiltration at the irradiated site; however, the TCR landscape remains dominated by polyclonal expansion of preexisting T-cell clones. Adaptive resistance via the PD-1/PD-L1 pathway restricts the generation of systemic anticancer immunity following radiotherapy, which can be overcome through combination with αPD-1 mAb leading to improved local and distal tumor control. Moreover, we show that effective clearance of tumor following combination therapy is dependent on both T cells resident in the tumor at the time of radiotherapy and infiltrating T cells.Conclusions: These data provide evidence that radiotherapy can enhance T-cell trafficking to locally treated tumor sites and augment preexisting anticancer T-cell responses with the capacity to mediate regression of out-of-field tumor lesions when delivered in combination with αPD-1 mAb therapy. Clin Cancer Res; 23(18); 5514-26. ©2017 AACR.


Innate Immune Responses to Highly Pathogenic Coronaviruses and Other Significant Respiratory Viral Infections.
The new pandemic virus SARS-CoV-2 emerged in China and spread around the world in <3 months, infecting millions of people, and causing countries to shut down public life and businesses. Nearly all nations were unprepared for this pandemic with healthcare systems stretched to their limits due to the lack of an effective vaccine and treatment. Infection with SARS-CoV-2 can lead to Coronavirus disease 2019 (COVID-19). COVID-19 is respiratory disease that can result in a cytokine storm with stark differences in morbidity and mortality between younger and older patient populations. Details regarding mechanisms of viral entry via the respiratory system and immune system correlates of protection or pathogenesis have not been fully elucidated. Here, we provide an overview of the innate immune responses in the lung to the coronaviruses MERS-CoV, SARS-CoV, and SARS-CoV-2. This review provides insight into key innate immune mechanisms that will aid in the development of therapeutics and preventive vaccines for SARS-CoV-2 infection.


The predictive value of early indicators for HBeAg seroconversion in HBeAg-positive chronic hepatitis B patients with Telbivudine treatment for 104 weeks.
Through an observation on HBeAg-positive chronic hepatits B (CHB) patients in Telbivudine (LDT) treatment for 104 weeks, we tried to explore valuable early predictors for HBeAg seroconversion during the treatment. A prospective study lasting for 104 weeks was conducted, and the patients enrolled were administered with LDT 600 mg daily. The medical evaluation went every 12 weeks, then the age distribution, baseline ALT level, early HBVDNA, HBsAg and HBeAg levels at baseline, week 12 and 24 as well as the decrease of the three indicators at week 12 and 24 were analyzed for their predictive values for HBeAg seroconversion at week 104. Thirty-three patients finished the observation. All patients got ALT normalisation and 28 patients (84.84%) got complete virological response (HBV DNA<291 copies/ml) at week 104. Poor virological response and virologic breakthrough was observed in two (6.06%) and three patients (9.09%), respectively. Nine patients (27.27%) got HBeAg seroconversion. HBeAg levels and its decrease levels at week 12 and 24 showed significant differences between patients with and without HBeAg seroconversion. And the HBsAg levels at week 12 and 24 showed tendencies of significant differences in two groups. HBeAg level at week 24 was confirmed related to its longer term seroconversion in regression analysis. The patients with HBeAg level<2.1 S/CO at week 24 would be more possible to get HBeAg seroconversion at week 104, with sensitivity, specificity, positive and negative predictive value of 95.83%, 88.89%, 95.8% and 88.9%, respectively. Good efficacy of long-term LDT treatment in biological and virological response and its advantage in serological response was confirmed again in our study. The HBeAg level at week 24 showed significant value in prediction for HBeAg seroconversion at week 104 compared to other serological markers in the early period.


Eugenol causes melanoma growth suppression through inhibition of E2F1 transcriptional activity.
Metastatic malignant melanoma is an extremely aggressive cancer, with no currently viable therapy. 4-Allyl-2-methoxyphenol (eugenol) was tested for its ability to inhibit proliferation of melanoma cells. Eugenol but not its isomer, isoeugenol (2-methoxy-4-propenylphenol), was found to be a potent inhibitor of melanoma cell proliferation. In a B16 xenograft study, eugenol treatment produced a significant tumor growth delay (p = 0.0057), an almost 40% decrease in tumor size, and a 19% increase in the median time to end point. More significantly, 50% of the animals in the control group died from metastatic growth, whereas none in the treatment group showed any signs of invasion or metastasis. Eugenol was well tolerated as determined by measurement of bodyweights. Examination of the mechanism of the antiproliferative action of eugenol in the human malignant melanoma cell line, WM1205Lu, showed that it arrests cells in the S phase of the cell cycle. Flow cytometry coupled with biochemical analyses demonstrated that eugenol induced apoptosis. cDNA array analysis showed that eugenol caused deregulation of the E2F family of transcription factors. Transient transfection assays and electrophoretic mobility shift assays showed that eugenol inhibits the transcriptional activity of E2F1. Overexpression of E2F1 restored about 75% of proliferation ability in cultures. These results indicate that deregulation of E2F1 may be a key factor in eugenol-mediated melanoma growth inhibition both in vitro and in vivo. Since the E2F transcription factors provide growth impetus for the continuous proliferation of melanoma cells, these results suggest that eugenol could be developed as an E2F-targeted agent for melanoma treatment.


Pilot study of store and forward teledermatology services in Perth, Western Australia.
In remote areas, telemedicine services can improve the quality of access to specialist medical care and dermatology is well suited to the use of this technology. There is no published work on teledermatology services in Australia. Our purpose was to investigate the reliability of dermatological diagnoses obtained using a store and forward telemedicine system, which is being developed to offer specialist consultative services to patients in remote areas of Western Australia. We report on a small prospective non-randomized pilot study conducted at Royal Perth Hospital, Western Australia which compared diagnoses reached following telemedicine consultations with diagnoses reached following traditional face-to-face consultations. In 25 out of 50 consultations, identical diagnoses were reached. In the remaining five cases, the preferred diagnosis and first differential diagnosis were reversed in order of preference. We feel this system is sufficiently promising to trial more extensively in the field.


Determination of xanthine in the presence of hypoxanthine by adsorptive stripping voltammetry at the mercury film electrode.
A stripping method for the determination of xanthine in the presence of hypoxanthine at the submicromolar concentration levels is described. The method is based on controlled adsorptive accumulation at the thin-film mercury electrode followed by a fast linear scan voltammetric measurement of the surface species. Optimum experimental conditions were found to be the use of 1.0 × 10(-3) mol L(-1) NaOH solution as supporting electrolyte, an accumulation potential of 0.00 V for xanthine and -0.50 V for hypoxanthine-copper, and a linear scan rate of 200 mV second(-1). The response of xanthine is linear over the concentration ranges of 20-140 ppb. For an accumulation time of 30 minutes, the detection limit was found to be 36 ppt (2.3 × 10(-10) mol L(-1)). Adequate conditions for measuring the xanthine in the presence of hypoxanthine, copper and other metals, uric acid, and other nitrogenated bases were also investigated. The utility of the method is demonstrated by the presence of xanthine associated with hypoxanthine, uric acid, nitrogenated bases, ATP, and ssDNA.


Voluntary control of agonist premotor silence preceding limb movements of maximal effort.
The ability to gain voluntary control over agonist premotor silence through electromyographic (EMG) feedback was examined in healthy subjects performing maximal horizontal elbow flexions. Subjects exhibiting premotor silence on at least 50% of the pretest trials showed significantly greater peak angular velocity than subjects who produced the silent period on fewer than 20% of the trials during the pretest. The latter subjects acquired control of agonist premotor silence with practice and graphic feedback regarding their EMG patterns. The subjects who were the most successful in learning to produce the silent period increased their angular velocity to the level of the subjects who naturally exhibited the inhibition. The less successful subjects showed smaller increases in velocity. The data provide further evidence that premotor silence is primarily under central influence, that its control can be acquired, and that it may be functionally related to contractile rate.


Platypnea-orthodeoxia syndrome in the right lateral decubitus position: a case report.
Platypnea-orthodeoxia syndrome is a rare syndrome characterized by dyspnea and hypoxia when the patient is sitting or standing. Here we report a case of platypnea-orthodeoxia syndrome caused by a right hemidiaphragmatic elevation with giant liver cyst that triggered a right-to-left shunt through the patent foramen ovale. This case report is the first presentation of a case secondary to hemidiaphragmatic elevation with giant liver cyst. In addition to this, a malposition of the pacemaker lead could be associated with platypnea-orthodeoxia syndrome in this case. A 91-year-old Japanese woman presented to our hospital with hypoxia of unknown origin. Severe hypoxia and cyanosis were observed only in the right lateral decubitus position. A chest X-ray and computed tomography scan revealed right hemidiaphragmatic elevation, which was probably compressing the right atrium. A transesophageal echocardiogram showed a compressed right atrium and shunt blood flow in both directions: from the left to the right atrium and vice versa. The shunt flow was exacerbated by postural changes from the left to the right lateral decubitus. A transesophageal echocardiogram also confirmed compression of the right atrium due to giant liver cyst and a malposition of the pacemaker lead abnormally placed in the left atrium through patent foramen ovale. We concluded that the cause of hypoxia was platypnea-orthodeoxia syndrome with right-to-left interatrial shunt through patent foramen ovale. Surgical closure of patent foramen ovale was not performed due to the age of our patient, surgical difficulties, and failure to obtain informed consent. For these reasons she was discharged after receiving medical advice about her posture. Platypnea-orthodeoxia syndrome is rare and difficult to diagnose. The present case suggests that hypoxia due to postural changes should be considered a differential diagnosis of platypnea-orthodeoxia syndrome.


Direct autofluorescence during CO2 laser surgery of the larynx: can it really help the surgeon?
Herein we assessed the impact of direct autofluorescence during intraoperative work-up on obtaining superficial free resection margins, identifying new areas of malignant transformation and altering disease-free survival and local control at 3 years in patients submitted to transoral laser surgery (TLS) for early glottic cancer. Prospective cohort evaluation was carried out on the diagnostic accuracy of the superficial extent and TNM staging in 73 patients with glottic carcinoma undergoing transoral CO2 laser surgery. The use of direct autofluorescence was associated with superficial disease-free margins in 97.2% of cases, and with superficial close margins in 2.8%. The improvement in diagnostic accuracy was 16.4%; in 8.2% of cases, there was upstaging of the TNM classification (in one case, a second neoplastic area in a different laryngeal site was observed and considered to be a second endolaryngeal primary). The sensitivity of direct autofluorescence was 96.5% with a specificity of 98.5%. Overall, 3-year disease-specific survival and local control with laser alone were, respectively: T1a (97.5%, 100%), T1b (86.7%, 86.7%), T2 (88.9%, 88.9%). This study demonstrates that direct autofluorescence can help to identify positive superficial margins, and has a favourable impact on disease-specific survival and local control at 3 years.


PPARδ, a Potential Therapeutic Target for Heart Disease.
The nuclear receptor peroxisome proliferator-activated receptor δ (PPARδ) can transcriptionally regulate target genes. PPARδ exerts essential regulatory functions in the heart, which requires constant energy supply. PPARδ plays a key role in energy metabolism, controlling not only fatty acid (FA) and glucose oxidation, but also redox homeostasis, mitochondrial biogenesis, inflammation, and cardiomyocyte proliferation. PPARδ signaling is impaired in the heart under various pathological conditions, such as pathological cardiac hypertrophy, myocardial ischemia/reperfusion, doxorubicin cardiotoxicity and diabetic cardiomyopathy. PPARδ deficiency in the heart leads to cardiac dysfunction, myocardial lipid accumulation, cardiac hypertrophy/remodeling and heart failure. This article provides an up-today overview of this research area and discusses the role of PPARδ in the heart in light of the complex mechanisms of its transcriptional regulation and its potential as a translatable therapeutic target for the treatment of cardiac disorders.


Intergenerational effects on fertility in male and female mice after chronic exposure to environmental doses of NSAIDs and 17α-ethinylestradiol mixtures.
Non-steroidal anti-inflammatory drugs (NSAIDs) and 17α-ethinylestradiol (EE2) are extensively used in human and veterinary medicine. Due to their partial removal by wastewater treatment plants, they are frequent environmental contaminants, particularly in drinking water. Here, we investigated the adverse outcomes of chronic exposure to mixtures of NSAIDs (ibuprofen, 2hydroxy-ibuprofen, diclofenac) and EE2 at two environmentally relevant doses in drinking water, on the reproductive organ development and fertility in F1-exposed male and female mice and in their F2 offspring. In male and female F1 mice, which were exposed to these mixtures, reproductive organ maturation, estrous cyclicity, and spermiogenesis were altered. These defects were observed also in F2 animals, in addition to some specific sperm parameter alterations in F2 males. Transcriptomic analysis revealed significant changes in gene expression patterns and associated pathways implicated in testis and ovarian physiology. Chronic exposure of mice to NSAID and EE2 mixtures at environmental doses intergenerationally affected male and female fertility (i.e. total number of pups and time between litters). Our study provides new insights into the adverse effects of these pharmaceuticals on the reproductive health and will facilitate the implementation of a future regulatory environmental risk assessment of NSAIDs and EE2 for human health.


Corneal wound healing: holographic stress-test analysis.
Double-exposure holograms are taken of the patient's cornea after a heartbeat. The resulting stress test, or holographic interferogram, shows sharply demarcated fringe patterns outlining structurally weak areas in the cornea. Wound strength in postoperative keratoplasties is now being evaluated on a routine basis to determine the effect of topical medications and surgical techniques. All grafts measured to date reveal "bulging" or weakness of the wound margin by holography even though clinically they are fully "healed." Only one patient had a vascularized region of the wound that was stronger than surrounding cornea. It is no longer necessary to resort to pulling the wound apart in animals for measurement of tensile strength.


Thickness of the adductor pollicis muscle in nutritional assessment of surgical patients.
To evaluate the correlation between thickness of the muscle adductor pollicis and anthropometric measurements, body mass index and Subjective Global Assessment in the nutritional assessment of surgical patients. The study population comprised patients admitted to the general and reconstructive surgery unit of a university hospital in the city of Vitória (ES), Brazil. The inclusion criteria were patients evaluated in the first 48 hours of admission, aged ≥20 years, hemodynamically stable, with no edema or ascites. Data analysis was performed using the software Statistical Package for Social Science 21.0, significance level of 5%. The sample consisted of 150 patients that were candidates to surgery, mean age of 42.7±12.0 years. The most common reasons for hospitalization were surgical procedures, gastrintestinal diseases and neoplasm. Significant association was observed between thickness of adductor pollicis muscle and Subjective Global Assessment (p=0.021) and body mass index (p=0.008) for nutritional risk. Significant correlation was found between thickness of adductor pollicis muscle and arm muscle circumference, corrected arm muscle area, calf circumference and body mass index. There were no significant correlations between thickness of adductor pollicis muscle and triceps skinfold and age. The use of thickness of adductor pollicis muscle proved to be an efficient method to detect malnutrition in surgical patients and it should be added to the screening process of hospitalized patients, since it is easy to perform, inexpensive and noninvasive.


Expression of inducible nitric oxide synthase in paired neoplastic and non-neoplastic primary prostate cell cultures and prostatectomy specimen.
Nitric oxide (NO) is an important signaling molecule for ischemia, inflammation, angiogenesis, immune response, and cell growth and differentiation. It has recently been shown that increased production of NO within various human cancers may contribute to tumor angiogenesis, tumor growth and metastasis, and tumor-related immune suppression. NO can be produced by several NO synthases (NOS), including inducible synthase (iNOS), which is expressed during cell activation and produces NO in larger quantity and for a longer period of time than non-inducible NOSs. In this study, we examined the expression levels of iNOS mRNA and protein in prostate adenocarcinoma using a paired nonneoplastic and neoplastic primary prostate cell culture system and related prostatectomy specimens. Six pairs of neoplastic and nonneoplastic primary prostate cell cultures were established from radical prostatectomy specimens based on homogeneity of the originating tumor and the nonneoplastic tissue. Radioactive reverse transcriptase polymerase chain reaction and subsequent quantitative analysis of iNOS mRNA were performed on the cultures using beta-actin as an internal control. Immunohistochemical studies with an anti-iNOS monoclonal antibody were performed on the corresponding formalin-fixed paraffin-embedded prostatectomy tissue sections. We observed marked patient-to-patient variation in "normal" levels of iNOS mRNA. However, all six neoplastic cultures showed moderately to markedly higher mRNA levels than did their paired nonneoplastic cultures. In addition, iNOS protein levels were significantly higher in paraffin-embedded prostate cancer tissue sections than in adjacent nonneoplastic tissue. Overexpression of iNOS mRNA and protein levels is present in moderately differentiated prostate adenocarcinoma and may contribute to prostate cancer angiogenesis, tumor growth, and tumor-related immunosuppression.


Annulate lamellae play only a minor role in the storage of excess nucleoporins in Drosophila embryos.
The nuclear pore complexes (NPCs), multiprotein assemblies embedded in the nuclear envelope, conduct nucleo-cytoplasmic traffic of macromolecules. Mimics of NPCs, called annulate lamellae pore complexes (ALPCs), are usually found in cytoplasmic membranous stacks in oocytes and early embryonic cells. They are believed to constitute storage compartments for excess premade nucleoporins. To evaluate the extent to which ALPCs store nucleoporins in early embryonic cells we took advantage of syncytial Drosophila embryos, containing both AL and rapidly proliferating nuclei in the common cytoplasm. Electron microscopic morphometric analysis showed that the number of ALPCs did not decrease to compensate for the growing number of NPCs during syncytial development. We performed Western blot analysis to quantify seven different nucleoporins and analyzed their intraembryonal distribution by confocal microscopy and subcellular fractionation. Syncytial embryos contained a large maternally contributed stockpile of nucleoporins. However, even during interphases, only a small fraction of the excess nucleoporins was assembled into ALPCs, whereas the major fraction was soluble and contained at least one phosphorylated nucleoporin. We conclude that in Drosophila embryos ALPCs play only a minor role in storing the excess maternally contributed nucleoporins. Factors that may prevent nucleoporins from assembly into ALPCs are discussed.


Impact of endplate-implant area mismatch on rates and grades of subsidence following stand-alone lateral lumbar interbody fusion: an analysis of 623 levels.
Stand-alone lateral lumbar interbody fusion (LLIF) is a useful minimally invasive approach for select spinal disorders, but implant subsidence may occur in up to 30% of patients. Previous studies have suggested that wider implants reduce the subsidence rate. This study aimed to evaluate whether a mismatch of the endplate and implant area can predict the rate and grade of implant subsidence. The authors conducted a retrospective review of prospectively collected data on consecutive patients who underwent stand-alone LLIF between July 2008 and June 2015; 297 patients (623 surgical levels) met inclusion criteria. Imaging studies were examined to grade graft subsidence according to Marchi criteria. Thirty patients had radiographic evidence of implant subsidence. The endplates above and below the implant were measured. A total of 30 patients with implant subsidence were identified. Of these patients, 6 had Marchi grade 0, 4 had grade I, 12 had grade II, and 8 had grade III implant subsidence. There was no statistically significant correlation between the endplate-implant area mismatch and subsidence grade or incidence. There was also no correlation between endplate-implant width and length mismatch and subsidence grade or incidence. However, there was a strong correlation between the usage of the 18-mm-wide implants and the development of higher-grade subsidence (p = 0.002) necessitating surgery. There was no significant association between the degree of mismatch or Marchi subsidence grade and the presence of postoperative radiculopathy. Of the 8 patients with 18-mm implants demonstrating radiographic subsidence, 5 (62.5%) required reoperation. Of the 22 patients with 22-mm implants demonstrating radiographic subsidence, 13 (59.1%) required reoperation. There was no correlation between endplate-implant area, width, or length mismatch and Marchi subsidence grade for stand-alone LLIF. There was also no correlation between either endplate-implant mismatch or Marchi subsidence grade and postoperative radiculopathy. The data do suggest that the use of 18-mm-wide implants in stand-alone LLIF may increase the risk of developing high-grade subsidence necessitating reoperation compared to the use of 22-mm-wide implants.


Structural and thermodynamic stability of uranyl-deferiprone complexes and the removal efficacy of U(vi) at the cellular level.
Deferiprone (3-hydroxy-1,2-dimethyl-4(1H)-pyridone, DFP), which is a drug clinically used for removing heavy metals in vivo, was explored for its removal efficiency towards uranium. The reaction of uranyl nitrate hexahydrate with DFP at room temperature yielded the compound [(UO2)(H2O)(C7NO2H8)2]·4H2O (1), which crystallizes from a mixed solution of methanol and water (pH = 7.0). X-ray diffraction shows that the stable complexation of uranyl occurs from the coordination of two bidentate DFP ligands perpendicular to the O[double bond, length as m-dash]U[double bond, length as m-dash]O unit with a fifth coordinating oxygen atom coming from one water molecule, resulting in a pentagonal bipyramidal geometry. The formation constants of uranyl and DFP complexes were measured and the species distribution diagram illustrates that UO2L2 (94.6%) is the dominant uranyl-DFP complex in 0.1 M KCl solution at physiological pH = 7.4. The results from both crystallographic and potentiometric studies imply that the metal : ligand ratio is 1 : 2. The effectiveness of using DFP to remove uranium was examined at the cellular level, and the results suggest that it can significantly reduce the cellular uptake and increase the cellular release of U(vi) in renal proximal tubular epithelial cells (NRK-52E).


Biological Movement and Laws of Physics.
Living systems may be defined as systems able to organize new, biology-specific, laws of physics and modify their parameters for specific tasks. Examples include the force-length muscle dependence mediated by the stretch reflex, and the control of movements with modification of the spatial referent coordinates for salient performance variables. Low-dimensional sets of referent coordinates at a task level are transformed to higher-dimensional sets at lower hierarchical levels in a way that ensures stability of performance. Stability of actions can be controlled independently of the actions (e.g., anticipatory synergy adjustments). Unintentional actions reflect relaxation processes leading to drifts of corresponding referent coordinates in the absence of changes in external load. Implications of this general framework for movement disorders, motor development, motor skill acquisition, and even philosophy are discussed.


[Bronchial asthma: characteristics of modern diagnostics].
Bronchial asthma, including its not severe forms, is chronic inflammatory disease of bronchial tubes, requiring application of anti-inflammatory therapy, foremost the use corticosteroids, which are capable to improve considerably prognosis for such patients. At the same to diagnose bronchial asthma is not easy task and presents some difficulty. The questions of correct diagnostics and treatment of this disease, its aspirin-sensitive form, and also such associated with bronchial asthma illnesses as bronchopulmonary aspergillosis, Churg-Strauss syndrome are discussed in the article. The role of gastrooesophageal reflux in forming of bronchial constriction is shown.


[The influence of empty-bed days on the median hospital stay in internal medicine].
To evaluate the epidemiology of delays in patients hospitalized in a department of internal medicine in a hospital of third level (high technology, end-stem of Spanish health system), its influences in hospital length of stay and the leading reasons which we named Gap Days. We studied all patients admitted through emergency ward for internal medicine during Oct 93-June 94. Gap Day was defined as the day passed as inpatient in which no intravenous route, isolation, artificial feeding, fever, impairing of clinic steady-state were needed or waited and any diagnostic tools were used. We counted Gap Day from the second day and from de third day for histopathology that we ordered the explorations. In a nonselected group days of delay to arrive written data were measured while the results were known for personal request. 144 patients had a mean length of stay of 9.52 (SD 5.41) days. Gap Days occurred in 97 (67%) patients (Mean 3.85 SD 2.80) with a mean length of stay 10.71 SD 5.09 days, while patients without Gap Days had a mean length of stay of 7.14 SD 5.29 days (p < 0.001). Patients with higher Gap Days were those with symptoms related to hematological system (p = 0.002), nephrourological system (p = 0.011) and a hematological diagnostic (p = 0.003) on admission. On discharge patients with hematological diagnostic had also higher Gap Days (p = 0.017). They had higher Gap Days also patients with two symptoms or more on admission (3.63 SD 2.96, p = 0.016), patients who lived alone (5.33 SD 3.42, p = 0.050) and patients with no concordance between diagnostic on admission and discharge (4.06 SD 3.41, p < 0.01). In 37 patients written data arrived 2.14 SD 1.06 days later after to know the results for personal request. Gap Days are an important factor to prolong the length of stay in internal medicine. They are influenced by number of symptoms on admission, concordance between diagnostics on admission and discharge, hematological diagnostics and some social and functioning hospital factors.


10-year experience with prenatal intervention for hydronephrosis.
We evaluated clinical outcomes after attempted prenatal intervention in fetuses with hydronephrosis. We retrospectively reviewed the histories of 10 fetuses considered for prenatal intervention between 1984 and 1993. One female and 8 male fetuses had bilateral hydroureteronephrosis and profound or progressive oligohydramnios, and 1 male fetus had massive progressive unilateral hydronephrosis. Shunt placement was not recommended and it was refused by the parents in 1 case each. Amniotic shunting was attempted in 8 fetuses between 22 and 28 weeks of gestation. Shunts were successfully placed with decreased hydronephrosis and increased amniotic fluid volume in 4 cases, while shunt placement was not technically possible in the remainder. Two shunts that retracted intra-abdominally at birth required laparotomy for retrieval. Postnatally all patients with shunts had compromised renal function. Of the 4 patients in whom attempts were unsuccessful 3 had mildly diminished renal function and 1 died of nonrenal causes (intraabdominal sepsis) on day 16 of life. No patient with a functioning shunt had postnatal pulmonary problems, whereas 3 without successful intervention had mild respiratory compromise. No definite advantage was noted in the small number of fetuses that underwent successful shunting. Successful shunt placement did not prevent renal insufficiency. The relief of oligohydraminos may benefit pulmonary function in some patients. The high technical failure and complication rates of in utero intervention should be considered before proceeding.


Presence and phosphorylation of transcription factors in developing dendrites.
In screening amplified poly(A) mRNA from hippocampal dendrites and growth cones in culture to determine candidates for local translation, we found that select transcription factor mRNAs were present. We hypothesized that synthesis of transcription factor proteins within dendrites would provide a direct signaling pathway between the distal dendrite and the nucleus resulting in modulation of gene expression important for neuronal differentiation. To evaluate this possibility, radiolabeled amplified antisense RNA was used to probe slot blots of transcription factor cDNAs as well as arrayed blots of zinc finger transcription factors. The mRNAs encoding the cAMP response element binding protein (CREB), zif 268, and one putative transcription factor were detected. We expanded upon these results showing that CREB protein is present in dendrites, that translation of CREB mRNA in isolated dendrites is feasible and that CREB protein found in dendrites can interact with the cis-acting cyclic AMP reponse element DNA sequence by using an in situ Southwestern assay. Further, CREB protein in dendrites is not transported to this site from the cell body because fluorescently tagged CREB microperfused into the soma did not diffuse into the dendrites. In addition, CREB protein microperfused into dendrites was rapidly transported to the nucleus, its likely site of bioactivity. Lastly, by using the isolated dendrite system we show that phosphorylation of Ser-133 on CREB protein can occur in isolated dendrites independent of the nucleus. These data provide a regulatory pathway in which transcription factors synthesized and posttranslationally modified in dendrites directly alter gene expression bypassing the integration of signal transduction pathways that converge on the nucleus.


Increased plasma MMP9 in integrin alpha1-null mice enhances lung metastasis of colon carcinoma cells.
Inhibitors of matrix metalloproteinases (MMPs) were developed as anticancer agents based on the observation that MMPs facilitate local tumor spread and metastasis by promoting matrix degradation and cell migration. Unfortunately, these inhibitors were unsuccessful in the clinical treatment of several cancers, including lung cancer. A possible reason contributing to their failure is that MMP activity is critical for the generation of inhibitors of tumor angiogenesis, including angiostatin. Thus, MMPs might play opposing roles in tumor vascularization and invasion. To determine which effect of elevated MMP levels dominates in the progression of metastatic cancer, experimental lung metastasis assays were performed in integrin alpha1-null mice, a genetic model for increased plasma levels of MMP9 and MMP9-generated angiostatin (Pozzi et al., Proc. Natl. Acad. Sci. USA 2000;97:2202-7). We show that while the number of lung colonies in integrin alpha1-null mice was significantly increased compared to their wild-type counterparts, tumor volume was markedly reduced. In vivo treatment with the MMP inhibitor doxycycline resulted in a significant decrease in the number of lung colonies in both genotypes, but the tumors that formed were bigger and more vascularized. Increased tumor vascularization paralleled decreased plasma levels of MMP9 and consequent decreased angiostatin synthesis. These results demonstrate that while inhibition of MMPs prevents and/or reduces tumor invasion and lung metastasis, it has the paradoxical effect of increasing the size and vascularization of metastatic tumors due to decreased generation of inhibitors of endothelial cell proliferation. The continued growth of these large well-vascularized tumors may explain the poor efficacy of MMP inhibitors in lung cancer clinical trials.


Urban wild meat consumption and trade in central Amazonia.
The switch from hunting wild meat for home consumption to supplying more lucrative city markets in Amazonia can adversely affect some game species. Despite this, information on the amounts of wild meat eaten in Amazonian cities is still limited. We estimated wild meat consumption rates in 5 cities in the State of Amazonas in Brazil through 1046 door-to-door household interviews conducted from 2004 to 2012. With these data, we modeled the relationship between wild meat use and a selection of socioeconomic indices. We then scaled up our model to determine the amounts of wild meat likely to be consumed annually in the 62 urban centers in central Amazonia. A total of 80.3% of all interviewees reported consuming wild meat during an average of 29.3 (CI 11.6) days per year. Most wild meat was reported as bought in local markets (80.1%) or hunted by a family member (14.9%). Twenty-one taxa were cited as consumed, mostly mammals (71.6%), followed by reptiles (23.2%) and then birds (5.2%). The declared frequency of wild meat consumption was positively correlated with the proportion of rural population as well as with the per capita gross domestic product of the municipality (administrative divisions) where the cities were seated. We estimated that as much as 10,691 t of wild meat might be consumed annually in the 62 urban centers within central Amazonia, the equivalent of 6.49 kg per person per year. In monetary terms, this amounts to US$21.72 per person per year or US$35.1 million overall, the latter figure is comparable to fish and timber production in the region. Given this magnitude of wild meat trade in central Amazonia, it is fundamental to integrate this activity into the formal economy and actively develop policies that allow the trade of more resilient taxa and restrict trade in species sensitive to hunting.


RAGE-induced changes in the proteome of alveolar epithelial cells.
The receptor for advanced glycation end-products (RAGE) is a pattern recognition receptor and member of the immunoglobulin superfamily. RAGE is constitutively expressed in the distal lung where it co-localizes with the alveolar epithelium; RAGE expression is otherwise minimal or absent, except with disease. This suggests RAGE plays a role in lung physiology and pathology. We used proteomics to identify and characterize the effects of RAGE on rat alveolar epithelial (R3/1) cells. LC-MS/MS identified 177 differentially expressed proteins and the PANTHER Classification System further segregated proteins. Proteins involved in gene transcription (RNA and mRNA splicing, mRNA processing) and transport (protein, intracellular protein) were overrepresented; genes involved in a response to stimulus were underrepresented. Immune system processes and response to stimuli were downregulated with RAGE knockdown. Western blot confirmed RAGE-dependent changes in protein expression for NFκB and NLRP3 that was functionally supported by a reduction in IL-1β and phosphorylated p65. We also assessed RAGE's effect on redox regulation and report that RAGE knockdown attenuated oxidant production, decreased protein oxidation, and increased reduced thiol pools. Collectively the data suggest that RAGE is a critical regulator of epithelial cell response and has implications for our understanding of lung disease, specifically acute lung injury. In the present study, we undertook the first proteomic evaluation of RAGE-dependent processes in alveolar epithelial cells. The alveolar epithelium is a primary target during acute lung injury, and our data support a role for RAGE in gene transcription, protein transport, and response to stimuli. More over our data suggest that RAGE is a critical driver of redox regulation in the alveolar epithelium. The conclusions of the present work assist to unravel the molecular events that underlie the function of RAGE in alveolar epithelial cells and have implications for our understanding of RAGE signaling during lung injury. Our study was the first proteomic comparison showing the effects of RAGE activation from alveolar epithelial cells that constitutively express RAGE and these results can affect a wide field of lung biology, pulmonary therapeutics, and proteomics.


Role of factor VIII C2 domain in factor VIII binding to factor Xa.
Factor VIII (FVIII) is activated by proteolytic cleavages with thrombin and factor Xa (FXa) in the intrinsic blood coagulation pathway. The anti-C2 monoclonal antibody ESH8, which recognizes residues 2248-2285 and does not inhibit FVIII binding to von Willebrand factor or phospholipid, inhibited FVIII activation by FXa in a clotting assay. Furthermore, analysis by SDS-polyacrylamide gel electrophoresis showed that ESH8 inhibited FXa cleavage in the presence or absence of phospholipid. The light chain (LCh) fragments (both 80 and 72 kDa) and the recombinant C2 domain dose-dependently bound to immobilized anhydro-FXa, a catalytically inactive derivative of FXa in which dehydroalanine replaces the active-site serine. The affinity (K(d)) values for the 80- and 72-kDa LCh fragments and the C2 domain were 55, 51, and 560 nM, respectively. The heavy chain of FVIII did not bind to anhydro-FXa. Similarly, competitive assays using overlapping synthetic peptides corresponding to ESH8 epitopes (residues 2248-2285) demonstrated that a peptide designated EP-2 (residues 2253-2270; TSMYVKEFLISSSQDGHQ) inhibited the binding of the C2 domain or the 72-kDa LCh to anhydro-FXa by more than 95 and 84%, respectively. Our results provide the first evidence for a direct role of the C2 domain in the association between FVIII and FXa.


Infection of chimpanzees with Nigerian I/CDC strain of Plasmodium ovale.
Seven splenectomized chimpanzees were infected with the Nigerian I/CDC strain of Plasmodium ovale. Two of the animals had no history of previous malarial infection whereas three had been infected with P. vivax, one with P. malariae, and one with P. vivax and P. malariae. The two animals with no previous malarial experience had maximum parasitemias of 88,700 and 127,000 per mm3 while the other animals had maximum parasitemias ranging from 10,100 to 60,600 per mm3. Anopheles freeborni, An. dirus, An. stephensi, and An. gambiae were readily infected via membrane feeding on heparinized blood obtained from these chimpanzees during the ascending phases of their primary attacks. The parasitemias in the chimpanzees with previous malarial experience were transient.


The significance of job-anxiety in a working population.
Job-anxiety, as distinguished from trait-anxiety, is associated with long-term sickness absence. The prevalence of job-anxiety within a working population is not known. Identifying individuals who would benefit from intervention might be useful. To investigate job-anxiety in employees not undergoing treatment for mental health illness, firstly by assessing the level of job-anxiety and work-related avoidance tendencies in a working sample, and secondly by testing whether job-anxiety is distinguishable from trait-anxiety. Cross-sectional survey of a convenience sample obtained through personal contact distribution. Employees from different professional settings completed an anonymous questionnaire and provided information on their employment status. The State-Trait-Anxiety Inventory (STAI-T) was used to measure trait-anxiety and the Job-Anxiety-Scale (JAS) was used to assess job (state) anxiety. There was a 69% response rate (240 responses); 188 responses were available for analysis of whom 62% were women. There were no employees with high trait-anxiety. Ten employees (5%) reported increased job-anxiety and of these nine employees reported high 'tendencies of avoidance and workplace absence'. Avoidance was most often accompanied by the comorbid job-anxieties 'job-related social anxiety', 'fear of changes at work' and 'fears of existence', 'anticipatory' and 'conditioned' job-anxiety and 'panic symptoms'. In this sample, self-reported job-anxiety appeared as a specific type of anxiety as opposed to trait-anxiety. In the workplace job-anxiety can present as job-avoidance and sickness absence and should be distinguished from trait-anxiety. In practice, employers and occupational health practitioners should be aware of those employees prone to sickness absence.


Access to safe abortion: building choices for women living with HIV and AIDS.
In many areas of the world where HIV prevalence is high, rates of unintended pregnancy and unsafe abortion have also been shown to be high. Of all pregnancies worldwide in 2008, 41% were reported as unintended or unplanned, and approximately 50% of these ended in abortion. Of the estimated 21.6 million unsafe abortions occurring worldwide in 2008 (around one in 10 pregnancies), approximately 21.2 million occurred in developing countries, often due to restrictive abortion laws and leading to an estimated 47,000 maternal deaths and untold numbers of women who will suffer long-term health consequences. Despite this context, little research has focused on decisions about and experiences of women living with HIV with regard to terminating a pregnancy, although this should form part of comprehensive promotion of sexual and reproductive health rights.In this paper, we explore the existing evidence related to global and country-specific barriers to safe abortion for all women, with an emphasis on research gaps around the right of women living with HIV to choose safe abortion services as an option for dealing with unwanted pregnancies. The main focus is on the situation for women living with HIV in Brazil, Namibia and South Africa as examples of three countries with different conditions regarding women's access to safe legal abortions: a very restrictive setting, a setting with several indications for legal abortion but non-implementation of the law, and a rather liberal setting.Similarities and differences are discussed, and we further outline global and country-specific barriers to safe abortion for all women, ending with recommendations for policy makers and researchers.


A mixed effects model for the analysis of repeated measures cross-over studies.
A mixed effects model is developed for cross-over trials in which the response is measured repeatedly within each time period. Relative to previous work on repeated measures cross-overs, the methodology synthesizes two important features. First, our procedure eliminates preliminary testing for carry-over, defined loosely as the component of a response that is due to treatment in the preceding period. This is achieved by generalizing the methodology to cross-over designs in which preliminary testing for carry-over is unnecessary. We focus largely on 'simple' carry-over, that is, carry-over that lasts for exactly one period and is independent of the treatment administered in the period in which the carry-over occurs. However, we also illustrate a modification of the procedure for a repeated measures cross-over design which uses a more complicated model of carry-over. Second, the model allows both the between- and within-subject variance to differ among treatments. Conditions are described wherein closed-form (CF) solutions to the variance components as well as closed-form hypothesis tests of the treatment differences exist. Flexibility in the model is illustrated with an example in which inference based on the CF likelihood-based estimates of the variance, and estimates formed using an iterative routine (PROC MIXED) are compared.


Thrombectomy for late occlusion of high flow extracranial-intracranial saphenous vein bypass graft after 27 years of patency.
High flow extracranial-intracranial (EC-IC) bypass with a saphenous vein graft (SVG) has been used for more than 40 years in patients with giant aneurysms of the posterior circulation refractory to medical management, and has demonstrated high long term patency rates. We report the case of a patient treated with external carotid artery (ECA)-posterior cerebral artery SVG bypass in 1989 who presented 27 years later with paresthesias and confusion, and was found to have partial occlusion of her SVG bypass graft and a basilar occlusion. She was treated with mechanical thrombectomy of the basilar occlusion via the partially thrombosed graft, the first report of such a procedure through a high flow posterior circulation EC-IC SVG, resulting in improvement of the patient's neurologic examination. At 27 years, this is the longest reported delay in thrombosis of a high flow SVG bypass graft, highlighting the long term patency of these grafts and the feasibility of thrombectomy through occluded bypass grafts.


Prediction of novel Bag-1 homologs based on structure/function analysis identifies Snl1p as an Hsp70 co-chaperone in Saccharomyces cerevisiae.
Polypeptide binding by the chaperone Hsp70 is regulated by its ATPase activity, which is itself regulated by co-chaperones including the Bag domain nucleotide exchange factors. Here, we tested the functional contribution of residues in the Bag domain of Bag-1M that contact Hsp70. Two point mutations, E212A and E219A, partially reduced co-chaperone activity, whereas the point mutation R237A completely abolished activity in vitro. Based on the strict positional conservation of the Arg-237 residue, several Bag domain proteins were predicted from various eukaryotic genomes. One candidate, Snl1p from Saccharomyces cerevisiae, was confirmed as a Bag domain co-chaperone. Snl1p bound specifically to the Ssa and Ssb forms of yeast cytosolic Hsp70, as revealed by two-hybrid screening and co-precipitations from yeast lysate. In vitro, Snl1p also recognized mammalian Hsp70 and regulated the Hsp70 ATPase activity identically to Bag-1M. Point mutations in Snl1p that disrupted the conserved residues Glu-112 and Arg-141, equivalent to Glu-212 and Arg-237 in Bag-1M, abolished the interaction with Hsp70 proteins. In live yeast, mutated Snl1p could not substitute for wild-type Snl1p in suppressing the lethal defect caused by truncation of the Nup116p nuclear pore component. Thus, Snl1p is the first Bag domain protein identified in S. cerevisiae, and its interaction with Hsp70 is essential for biological activity.


Needlescopic versus conventional laparoscopic surgery for colorectal cancer ~a comparative study~.
This study set out to determine whether Needlescopic surgery (NS) produces comparable surgical outcomes for patients with colorectal cancer (CRC) compared to conventional multi-port laparoscopic surgery (MPS). We used the five-port method with a 3.5 cm umbilical incision for extraction and reconstruction during MPS for CRC. One or two 5 mm ports were exchanged for needle forceps and all surgical procedures were as for previous MPS since July 2012. We investigated the short-term outcomes of 138 consecutive patients who underwent curative resection of CRC by NS (July 2012-August 2014) and 130 consecutive patients with CRC treated with MPS during a previous period (January 2010-June 2012). Operative time in the NS group was comparable to that of MPS (p=0.467); the NS group had significantly less estimated blood loss (p=0.002) and a shorter postoperative hospital stay (p<0.001). The mean number of dissected lymph nodes was 27 in both groups (p=0.730). No mortality occurred in either group, and similar morbidity rates were observed (p=0.454). NS using Endo Relief needle forceps is a safe and feasible option compared to conventional MPS for CRC.


[Nutrition and levels of risk factors for ischemic heart disease in 30- to 59-year-old men of Novosibirsk and resident newcomers of Chukotka].
Data on actual diets and major coronary risk factors are presented for residents of a district in Novosibirsk and nonaboriginal residents of Chukotka. Epidemiologic (quantitative assessment of dietary habits, questioning to detect angina of effort), biochemical (assays of plasma cholesterol, triglycerides, high density lipoprotein cholesterol) and physiologic (arterial BP, height and weight measurements) methods were used. Differences are demonstrated between the diets common for Novosibirsk and nonaboriginal Chukotka residents, which show correlation to differences in the incidence of major coronary risk factors.


[Regional metastases of squamous cell carcinoma of the lower lip].
Lower lip squamous cell carcinoma is the most common oral carcinoma. In relation to other oral carcinomas, it has a better clinical course. If regional metastases appear, lower lip carcinoma has a very poor prognosis. This study included patients treated for lower lip squamous cell carcinoma in the period 1984-2001. We examined neck metastases and time of their appearance after operation of the primary neoplasm. Our retrospective study included 305 patients with lower lip carcinoma in a twelve-year period. Out of this number, 293 had no positive neck lymph nodes, and only 12 patients had (4%). More than 75% of metastases appeared in the first year after surgery of the primary neoplasm. Contra-lateral metastases were most common. Taking into consideration the percentage of metastases, particularly during the first year after surgery, we suggest identification of sentinel lymph nodes and depending on PH finding one should decide about the type of neck operation. Frequent check-ups are necessary, especially during the first year.


Patterns of Gene Expression in Western Corn Rootworm (Diabrotica virgifera virgifera) Neonates, Challenged with Cry34Ab1, Cry35Ab1 and Cry34/35Ab1, Based on Next-Generation Sequencing.
With Next Generation Sequencing technologies, high-throughput RNA sequencing (RNAseq) was conducted to examine gene expression in neonates of Diabrotica virgifera virgifera (LeConte) (Western Corn Rootworm, WCR) challenged with individual proteins of the binary Bacillus thuringiensis insecticidal proteins, Cry34Ab1 and Cry35Ab1, and the combination of Cry34/Cry35Ab1, which together are active against rootworm larvae. Integrated results of three different statistical comparisons identified 114 and 1300 differentially expressed transcripts (DETs) in the Cry34Ab1 and Cry34/35Ab1 treatment, respectively, as compared to the control. No DETs were identified in the Cry35Ab1 treatment. Putative Bt binding receptors previously identified in other insect species were not identified in DETs in this study. The majority of DETs (75% with Cry34Ab1 and 68.3% with Cry34/35Ab1 treatments) had no significant hits in the NCBI nr database. In addition, 92 DETs were shared between Cry34Ab1 and Cry34/35Ab1 treatments. Further analysis revealed that the most abundant DETs in both Cry34Ab1 and Cry34/35Ab1 treatments were associated with binding and catalytic activity. Results from this study confirmed the nature of these binary toxins against WCR larvae and provide a fundamental profile of expression pattern of genes in response to challenge of the Cry34/35Ab1 toxin, which may provide insight into potential resistance mechanisms.


Activities at the Universal Protein Resource (UniProt).
The mission of the Universal Protein Resource (UniProt) (http://www.uniprot.org) is to provide the scientific community with a comprehensive, high-quality and freely accessible resource of protein sequences and functional annotation. It integrates, interprets and standardizes data from literature and numerous resources to achieve the most comprehensive catalog possible of protein information. The central activities are the biocuration of the UniProt Knowledgebase and the dissemination of these data through our Web site and web services. UniProt is produced by the UniProt Consortium, which consists of groups from the European Bioinformatics Institute (EBI), the SIB Swiss Institute of Bioinformatics (SIB) and the Protein Information Resource (PIR). UniProt is updated and distributed every 4 weeks and can be accessed online for searches or downloads.


Clinical profile, outcomes, and progression to type 2 diabetes among Indian women with gestational diabetes mellitus seen at a diabetes center in south India.
To describe the clinical profile, maternal and fetal outcomes, and the conversion rates to diabetes in women with gestational diabetes mellitus (GDM) seen at a tertiary care diabetes center in urban south India. Clinical case records of 898 women with GDM seen between 1991 and 2011 were extracted from the Diabetes Electronic Medical Records (DEMR) of a tertiary care diabetes center in Chennai, south India and their clinical profile was analyzed. Follow-up data of 174 GDM women was available. To determine the conversion rates to diabetes, oral glucose tolerance test (OGTT) was done in these women. Glucose tolerance status postpartum was classified based on World Health Organization (WHO) 2006 criteria. The mean maternal age of the women was 29 ± 4 years and mean age of gestation at first visit were 24 ± 8.4 weeks. Seventy percent of the women had a family history of diabetes. Seventy-eight percent of the women delivered full-term babies and 65% underwent a cesarean section. The average weight gain during pregnancy was 10.0 ± 4.2 kg. Macrosomia was present in 17.9% of the babies, hypoglycemia in 10.4%, congenital anomalies in 4.3%, and the neonatal mortality rate was 1.9%. Mean follow-up duration of the 174 women of whom outcome data was available was 4.5 years. Out of the 174, 101 women who were followed-up developed diabetes, of whom half developed diabetes within 5 years and over 90%, within 10 years of the delivery. Progression to type 2 diabetes mellitus (T2DM) in Indian women with GDM is rapid. There is an urgent need to develop standardized protocols for GDM care in India that can improve the maternal and fetal outcomes and help prevent future diabetes in women with GDM.