Obese Mice with Dyslipidemia Exhibit Meibomian Gland Hypertrophy and Alterations in Meibum Composition and Aqueous Tear Production.
Dyslipidemia may be linked to meibomian gland dysfunction (MGD) and altered meibum lipid composition. The purpose was to determine if plasma and meibum cholesteryl esters (CE), triglycerides (TG), ceramides (Cer) and sphingomyelins (SM) change in a mouse model of diet-induced obesity where mice develop dyslipidemia. Male C57/BL6 mice (8/group, age = 6 wks) were fed a normal (ND; 15% kcal fat) or an obesogenic high-fat diet (HFD; 42% kcal fat) for 10 wks. Tear production was measured and meibography was performed. Body and epididymal adipose tissue (eAT) weights were determined. Nano-ESI-MS/MS and LC-ESI-MS/MS were used to detect CE, TG, Cer and SM species. Data were analyzed by principal component analysis, Pearson's correlation and unpaired t-tests adjusted for multiple comparisons; significance set at p ≤ 0.05. Compared to ND mice, HFD mice gained more weight and showed heavier eAT and dyslipidemia with higher levels of plasma CE, TG, Cer and SM. HFD mice had hypertrophic meibomian glands, increased levels of lipid species acylated by saturated fatty acids in plasma and meibum and excessive tear production. The majority of meibum lipid species with saturated fatty acids increased with HFD feeding with evidence of meibomian gland hypertrophy and excessive tearing. The dyslipidemia is associated with altered meibum composition, a key feature of MGD.


[Reorganization of a third-level maternity ward during the COVID-19 pandemic: maternity ward experience at the University Hospital of Marrakech].
COVID-19 pandemic has put unprecedented pressures on obstetrics and gynecology care services. Maternity hospitals have needed to rapidly prepare to provide quality care while preventing the transmission of the infection. The purpose of this study is to describe the key elements of the response to COVID-19 within the maternity ward of the Mohammed VI University Hospital in Marrakech. a case study was conducted. Data were collected using various administrative documents related to activities in the maternity ward of the Mohammed VI University Hospital during the COVID-19 pandemic. We also conducted semi-structured interviews with professionals at the maternity ward. Qualitative thematic analysis approach was used for the analysis of transcribed data and triangulation, with the analysis of documentary data. the study highlighted phases of design and organization of two maternity ward access circuits. A new circuit for suspicious or confirmed cases of COVID-19 ensured access to quality care for patients, by guaranteeing isolation measures. The access circuit that was commonly used remained functional to consolidate the right of access to tertiary obstetrics and gynecology care while applying protective measures against COVID-19. this study highlights the necessity to draw on the experience of other health facilities, to contextualize local activities and to anticipate work organization in the face of health crises.


Langerin-mediated internalization of a modified peptide routes antigens to early endosomes and enhances cross-presentation by human Langerhans cells.
The potential of the skin immune system to generate immune responses is well established, and the skin is actively exploited as a vaccination site. Human skin contains several antigen-presenting cell subsets with specialized functions. In particular, the capacity to cross-present exogenous antigens to CD8+ T cells is of interest for the design of effective immunotherapies against viruses or cancer. Here, we show that primary human Langerhans cells (LCs) were able to cross-present a synthetic long peptide (SLP) to CD8+ T cells. In addition, modification of this SLP using antibodies against the receptor langerin, but not dectin-1, further enhanced the cross-presenting capacity of LCs through routing of internalized antigens to less proteolytic early endosome antigen 1+ early endosomes. The potency of LCs to enhance CD8+ T-cell responses could be further increased through activation of LCs with the toll-like receptor 3 ligand polyinosinic:polycytidylic acid (pI:C). Altogether, the data provide evidence that human LCs are able to cross-present antigens after langerin-mediated internalization. Furthermore, the potential for antigen modification to target LCs specifically provides a rationale for generating effective anti-tumor or anti-viral cytotoxic T lymphocyte responses.


Interobserver reliability in quantitative MRI assessment of temporomandibular joint disk status.
The purpose of this study was to investigate interobserver reliability of a new technique for quantification of magnetic resonance images of temporomandibular joint disk status. Sixty magnetic resonance images of adolescent temporomandibular joints were randomly drawn for analysis. Four experienced observers traced the articular disk and osseous structures on sagittal magnetic resonance slice images. Quantitative measurements of disk length and disk displacement were recorded for each slice of 57 joints traced by each observer through use of a new quantification technique. Intraclass correlation coefficients were computed to assess interobserver agreement in the tracing of joint structures. The calculated intraclass correlation coefficient was 0.681 for disk length and 0.830 for disk displacement. In addition, the mean variability among observers was 1.041 mm for measurement of disk length and 0.972 mm for measurement of disk displacement. Interobserver agreement is high when the new quantification technique is used to interpret magnetic resonance images.


Identification of differences between the surface proteins and glycoproteins of normal mouse (Balb/c) and human erythrocytes.
The topography of the external surface of the Balb/c mouse erythrocyte has been investigated and compared to the human erythrocyte by using a series of protein radiolabeling probes. After sodium dodecyl sulfate-polyacrylamide gel electrophoresis, the pattern of Coomassie Blue stained proteins was very similar for mouse and human erythrocyte ghosts, as was the distribution of radioactivity in protein bands after lactoperoxidase catalyzed radioiodination. The mouse erythrocyte glycoproteins identified by periodic-acid-Schiff and 'Stains-All' reagents, sialic acid analysis of gel slices, binding of 125I-wheat germ agglutinin and 125I-concanavalin A to the gels, and glycoprotein radiolabeling techniques, differed markedly from the sets of proteins labeled by radioiodination, and also differed from the human erythrocyte glycoproteins. Instead of the PAS I to PAS IV series of sialoglycoproteins characteristic of human erythrocytes, the mouse erythrocyte possesses a broad band of sialoglycoproteins with several peaks ranging in mol wt from 65,000 to 32,000. The same group of sialoglycoproteins were labeled by the periodate/B3H4-technique specific for terminal sialic acid, and the galactose oxidase/B3H4-method (plus neuraminidase) specific for galactosyl/N-acetylgalactosaminyl residues penultimate to sialic acid. These results emphasize the necessity to employ a variety of protein radiolabeling probes based on different labeling specificities, to study the membrane topography of cells which are poorly understood compared to the human erythrocyte membrane.


Coinfection With HIV and SARS-CoV-2 in Wuhan, China: A 12-Person Case Series.
Given the magnitude of the global COVID-19 pandemic, persons living with HIV (PLWH) may become coinfected with SARS-CoV-2. We conducted a survey in Wuhan, China, to characterize the status of coinfected PLWH, their time to clinical improvement, and clinical prognoses. Using a Wuhan shipping service for antiretroviral medications, the Wuhan LGBT Center screened 2900 PLWH shipping addresses and cross-referenced 36 of them to quarantine sites or hospitals, suggesting possible COVID-19 cases. Through telephone calls and WeChat (social media) messaging, we conducted a survey after obtaining online informed consent. We had 12 HIV-infected respondents (10 men and 2 women) who also reported COVID-19. The median age was 36 years (interquartile range: 33.0-56.3), mean age 42.4 years, and range 25-66 years of age. Nine of 10 persons on antiretroviral therapy (ART) presented with only mild COVID-19 symptoms. The 10th person on ART was a 56-year-old man who died at home early in the outbreak when health care services were overwhelmed. Two additional cases who had been in intensive care with acute COVID-19 were both men, aged 25 and 37 years; both were ART-naive until this hospitalization. Excluding the deceased man, 6 of 11 coinfected persons reported feeling depressed even after clinical improvements. Twelve coinfected persons were identified in Wuhan; 9 of 10 were on long-term ART and had favorable outcomes. Two men identified as having started ART only recently were found to have severe symptoms. Our case series suggests the value of ART for potential mitigation of COVID-19 coinfection.


Membrane protein structure determination: back to the membrane.
NMR spectroscopy enables the structures of membrane proteins to be determined in the native-like environment of the phospholipid bilayer membrane. This chapter outlines the methods for membrane protein structural studies using solid-state NMR spectroscopy with samples of membrane proteins incorporated in proteoliposomes or planar lipid bilayers. The methods for protein expression and purification, sample preparation, and NMR experiments are described and illustrated with examples from OmpX and Ail, two bacterial outer membrane proteins that function in bacterial virulence.


Outcome after traumatic brain injury: effects of aging on recovery.
To identify differences in outcome after traumatic brain injury (TBI) compared with orthopedic injuries as a function of age. Longitudinal data analyses from an inception cohort. Outpatient rehabilitation program. Eighty-two orthopedic injury patients and 195 TBI patients. Not applicable. Independent living, employment, and level of functioning 1 to 2 years after injury. Older patients and those with TBI were more likely to have increased dependence postinjury. Older TBI patients were more likely to have changes in employment status compared with orthopedic injury patients younger or TBI. The Mayo-Portland Adaptability Inventory and Disability Rating Scale were moderately predictive of level of functioning, return to employment, and independent living status 1 to 2 years postinjury. Injury severity was only mildly predictive of outcome. The effect of age on outcome affects recovery from neurologic injuries and, to a lesser extent, orthopedic injuries. Outcome after TBI is best predicted by patients' age and estimates of level of function at discharge. Findings suggest that older patients and those with TBI have a greater likelihood of becoming physically and financially dependent on others. Rehabilitation efforts should focus on maximizing levels of independence to limit financial and emotional costs to patients and their families.


Pathway-specificity in N-methyl-D-aspartate receptor-mediated synaptic inputs onto subplate neurons.
The subplate plays an important role in forming neuronal connections during early cortical development. We characterized by the use of whole-cell and cell-attached patch-clamp recordings in coronal brain slices from newborn mice (postnatal day [P] 0-3) the functional properties of two major pathways onto subplate neurons (SPn), the thalamocortical and the intra-subplate synaptic input. The two afferent pathways were stimulated extracellularly with bipolar electrodes placed in the thalamus and the subplate, respectively. Synaptically evoked and pharmacologically isolated N-methyl-d-aspartate receptor (NMDAR) -mediated responses with an onset latency of approximately 6 ms could be reliably recorded in P0-3 SPn. Whereas the intra-subplate input revealed a pronounced facilitation using paired pulse stimulation at 60-120 ms or repetitive activation at 10-40 Hz, the thalamocortical input was either stable or markedly suppressed under these conditions. Single cell reverse transcription PCR revealed the expression of the NR2A, B and D subunit in all investigated SPn. The intra-subplate and the thalamocortical synaptic input did not differ in their sensitivity to NVP-AAM077 or ifenprodil, indicating that both synaptic inputs have a similar NR2A/2B subunit composition. At P0, NMDAR-mediated synaptic inputs arising from the thalamus were significantly larger as compared with the intra-subplate input. This difference could no longer be detected in P2-3 SPn, indicating an input-specific developmental regulation during the first Ps. Our data indicate that the thalamocortical and intra-subplate synaptic input onto P0-3 SPn differs in functional, molecular and developmental properties. The intra-subplate synaptic input shows more mature functional properties and sustains high stimulation frequencies, thereby promoting the immature thalamocortical input to the developing neocortical circuit.


[Psychophysiological health aspects of population living in radiation-contaminated territories].
A combined psychophysiologic study of 100 individuals living in territories contaminated by radionuclides and 60 people from ecologically clean region. There were no significant differences in the incidence of neurotic disorders between people from these regions (61.2 and 65.3%, respectively). The highest sensitivity to small radiation doses was observed at rather young age (20-40 years). It was found that a considerable role in the development of borderline psychic disorders belongs to both somatic disturbances due to low-dose radiation and psychogenic effects. A combined action of psychogenic and somatogenic factors aggravated symptoms of neurotic disorders and promoted widening of a range of psychic disturbances. Some part of the population from the contaminated territories tended to the development of cerebro-vascular psychoorganic pathology resultant from a breakdown of the reserves of the adaptation. Now the stress factor of radionuclides exposure is not so important as social and economic problems.


Histamine release by neurotensin in the rat hindquarter: structure-activity studies.
Histamine releasing effects of neurotensin (NT) and several NT fragments and structural analogues were measured in the rat perfused hindquarter. The results show that the chemical groups responsible for histamine release are located in the C-terminal sequence Arg9-Pro10-Tyr11-Ile12-Leu13-OH. Both the spatial configuration and positive charge of Arg8 and Arg9 appear to contribute to the histamine releasing effect of NT. Optimization of the histamine releasing effect of NT requires both a free C-terminal carboxyl group and the presence in position 11 of NT of an aromatic residue, with the L-configuration, bearing an heteroatom capable of hydrogen bonding with the receptor. The results indicate that the structural requirements of NT to induce histamine release from the rat perfused hindquarter are similar to those involved in other peripheral biological actions of NT.


The value of radionuclide scanning in early diagnosis of intestinal infarction.
Small focal areas of intestinal infarction were produced experimentally in mice by two techniques: (1) intussusception and (2) suture strangulation of a loop of jejunum. The radionuclide technetium 99m diphosphonate (99mTc-diphosphonate) was subsequently administered intravenously, and abdominal scans were obtained. The scans demonstrated focal infarcted lesions accurately, and every positive scan was associated with an area of infarction, as demonstrated by pathologic examination. However, negative scans did not definitely exclude an area of infarction: in the intussusception study there were five false negative scans in 25 animals, and in the loop ligation study there were two false negative scans in 28 animals. The technique may have clinical application in such intestinal diseases as necrotizing enterocolitis in infancy in which early diagnosis of small areas of necrosis is currently difficult by clinical and radiologic methods.


Carcinoma of colon after ureterocolic anastomosis: implantation on calyceal mucosa.
Two cases are reported of the development of mucoid adenocarcinoma of the colon in young patients, nineteen and thirty-four years after ureterocolic anastomosis for benign conditions. One patient showed implantation of the colonic neoplasm on the mucosa of an upper calyx in the obstructed kidney. Patients who have undergone ureterocolic anastomosis have a greatly increased risk of the development of large-bowel neoplasms. When suspected, radiologic examination of the colon using a water-soluble contrast material, together with rectosigmoidoscopy or colonoscopy, is essential. Because of the serious nature of this complication, it is suggested that the recent trend to return to the use of large bowel in urinary diversion may be hazardous, especially in the young patient with a benign disease.


Pubertal ovarian hormone exposure reduces the number of myelinated axons in the splenium of the rat corpus callosum.
The size of the female rat corpus callosum decreases in response to pubertal ovarian hormone exposure, but the underlying changes in axonal composition have not been examined. In the current study, animals underwent ovariectomy or sham surgery at day 20, and the number of myelinated and unmyelinated axons were examined in young adulthood (2 months) using electron microscopy. Ovariectomized animals had a greater number of myelinated axons compared to intact animals, while total axon number was not affected. Ovarian hormone exposure seems to limit the number of axons that become myelinated in the splenium, while not affecting the number of axons crossing through the region.


A comparison between the effects of low (1 microg) and standard dose (250 microg) ACTH stimulation tests on adrenal cortex functions with Behçet's disease.
Behçet's disease is a rare, chronic disorder. The cause of Behçet's disease is unknown. It is believed to be caused by an autoimmune reaction. As in other chronic autoimmune diseases, Behçet's disease may show a subclinical adrenal failure and some changes in cortisol levels. We aimed to evaluate adrenal gland function in Behçet's disease patients. This study included 18 Behçet's disease patients and 15 healthy controls. Patient and control groups were administered i.v. 1 microg low dose test (LDT) and 250 microg standard dose test (SDT) adrenocorticotropic hormone (ACTH) stimulation test after 12 h of night fasting with an interval of 3-days and cortisol responses in the 0th, 30th and 60th minutes were evaluated. There was no statistically significant difference between basal cortisol values of Behçet's disease and control groups. Cortisol values in the 60th minute in LDT were significantly lower in Behçet's disease group than in the control group. In the peak cortisol responses to LDT, a significant decrease was found in Behçet's disease group. These findings suggest that hypothalamo-pituitary adrenal axis is partially suppressed in Behçet's disease.


The life span of erythrocytes in iron-deficiency anaemia.
Red cells, derived from 13 patients with iron-deficiency anaemia, were found to have a normal life span in the patients' own circulations and those of normal volunteers.


Analyzing the Integrin Adhesome by In Situ Proximity Ligation Assay.
The in situ proximity ligation assay (PLA) is capable of detecting single protein events such as protein protein-interactions and posttranslational modifications (e.g., protein phosphorylation) in tissue and cell samples prepared for analysis by immunofluorescent or immunohistochemical microscopy. The targets are detected using two primary antibodies which must be from different host species. A pair of secondary antibodies (PLA probes) conjugated to complementary oligonucleotides is applied to the sample, and a signal is generated only when the two PLA probes are in close proximity by their binding to the two primary antibodies that have bound to their targets in close proximity. The signal from each pair of PLA probes is visualized as an individual fluorescent spot. These PLA signals can be quantified (counted) using image analysis software (ImageJ), and also assigned to a specific subcellular location based on microscopy image overlays. In principle, in situ PLA offers a relatively simple and sensitive technique to analyze interactions among any proteins for which suitable antibodies are available. Integrin-mediated focal adhesions (FAs) are large multiprotein complexes consisting of more than 150 proteins, also known as the integrin adhesome, which link the extracellular matrix (ECM) to the actin cytoskeleton and regulate the functioning of mechanosignaling pathways. The in situ PLA approach is well suited for examining the spatiotemporal aspects of protein posttranslational modifications and protein interactions occurring in dynamic multiprotein complexes such as integrin mediated focal adhesions.


Asymmetric oxidative Lewis base catalysis-unifying iminium and enamine organocatalysis with oxidations.
Enantioselective oxidative domino reactions of allylic alcohols to functionalized aldehydes have been developed. The one pot domino oxidation-iminium activation represents a convenient strategy for the enantioselective addition of malonates to allylic alcohols and the asymmetric formation of formyl cyclopropanes.


Reversion of 'early worsening' of diabetic retinopathy by deliberate restoration of poor metabolic control.
Acutely lowering long-standing severe hyperglycaemia can trigger progression ('early worsening') of diabetic retinopathy, most likely by up-regulation of the circulating insulin-like growth factor 1 (IGF-1). This condition, also called 'florid retinopathy', rarely responds to standard laser coagulation treatment. In this retrospective report, 2 young patients with type 2 diabetes are described, in whom deliberate restoration of poor diabetes control reduced the serum IGF-1 levels and improved 'early worsened' diabetic retinopathy.


The Isthmus of Panama: a major physical barrier to gene flow in a highly mobile pantropical seabird.
To further test the hypothesis that the Isthmus of Panama is a major barrier to gene flow in pantropical seabirds, we applied phylogeographic methods to mitochondrial control sequence variation in masked booby (Sula dactylatra) populations on either side of the Isthmus of Panama and the southern tip of Africa. In accord with Steeves et al. (2003), we found that all Caribbean masked boobies with the 'secondary contact' cytochrome b haplotype (m-B) shared a control region haplotype (Sd_100), which grouped with Indian-Pacific haplotypes and not Caribbean-Atlantic haplotypes. In addition, Sd_100 was more closely related to control region haplotypes in the Indian Ocean than in the Pacific. We also found that the 'secondary contact' birds diverged more recently from extant populations in the Indian Ocean than in the Pacific. Thus, it appears that these masked boobies did not breach the Isthmus of Panama. Rather, birds likely dispersed around the southern tip of Africa during favourable oceanographic conditions in the Pleistocene.


Examining the Role of Resilience, Posttraumatic Growth, and Quality of Life in Women with Breast Cancer: A Serial Multiple Mediator Model Approach.
It is unclear how resilience and posttraumatic growth help women with breast cancer face cancer-related symptom distress. This study included both resilience and posttraumatic growth as mediators in a serial multiple mediator model to examine changes in the relationship between symptom distress and quality of life among women with breast cancer. We conducted the descriptive, cross-sectional study in Taiwan. Data were collected using a survey that assessed symptom distress, resilience, posttraumatic growth, and quality of life. A serial multiple mediator model examined one direct and three specific indirect effects of symptom distress on quality of life through resilience and posttraumatic growth. All 91 participants reported the presence of symptom distress and moderate levels of resilience. Quality of life was significantly associated with symptom distress (b = -1.04), resilience (b = 0.18), and posttraumatic growth (b = 0.09). The indirect effect of symptom distress on quality of life through resilience alone was statistically significant (b = -0.23, 95% CI -0.44 to -0.07) and statistically greater than the specific indirect effect through resilience and posttraumatic growth combined (b = -0.21, 95% CI -0.40 to -0.05). Resilience plays a unique role in reducing the impact of symptom distress on the quality of life among women with breast cancer. Given the importance of resilience to quality of life, oncology nurses can assess the resilience of women with breast cancer and help identify available internal, external, and existential resources to strengthen their resilience.


Vital sign, or none of our business?
For the first time in the history of the NHS, the Department of Health has issued guidelines for a national smoking cessation policy in England. The recommendations include guidance to all health commissioners on the introduction of 'systems to maintain an up to date record of the smoking status of all patients in their notes. It should be regarded as a vital sign.'


The Relationship Between Polycystic Ovarian Syndrome, Periodontal Disease, and Osteoporosis.
Polycystic ovarian syndrome (PCOS) is the most common endocrine disorder to affect women in their reproductive years. There has been growing concern that women with PCOS may suffer from long-term metabolic consequences due to the high degree of insulin resistance that is often present in PCOS. However, few longitudinal studies exist in this area and there is a paucity of data on whether women with PCOS are at risk of other chronic diseases as they age. Specifically, current evidence suggests that reproductive-age women with PCOS may be at increased risk for both osteoporosis and periodontal disease (PD)-both these chronic diseases can have serious implications for health and quality of life. However, few studies have addressed how risk factors for osteoporosis and PD may be altered by aging in PCOS. The PCOS phenotype of women beyond reproductive years is poorly understood, and it is not known whether the metabolic profile of older women with PCOS results in an increased risk of osteoporosis and PD. The objective of this review is to discuss the relationships between PCOS, osteoporosis, and PD, and how these relationships could be impacted during aging. The long-term goal of this review is to provide direction for future research that is needed to more clearly elucidate these relationships and eventually provide a basis for evidence-based health recommendations.


Schisandrin B suppresses gastric cancer cell growth and enhances the efficacy of chemotherapy drug 5-FU in vitro and in vivo.
Gastric cancer (GC) is a serious affliction worldwide and remains to be the fourth most common cancer with poor prognosis, especially in advanced stage. Chemotherapy is one of the main therapeutic means. The purpose of this study was to investigate the antitumor effects of Schisandrin B (Sch B) on GC cells both in vitro and in vivo, as well as the synergistic effect with 5-fluorouracil (5-FU), and to preliminarily explore the relevant mechanism of action. Our results showed that Sch B inhibited the growth, migration and invasion of GC cells. Besides, Sch B could effectively inhibit the phosphorylation of STAT3 (signal transducer and activator of transcription 3), induce autophagy, and enhance the efficacy of chemotherapy drug 5-FU in vitro and in vivo. Taken together, the findings indicate that Sch B displays potent antitumor activities. The co-administration of Sch B and 5-FU might be a promising way for future therapy of GC.


Successful Conservative Management of Inferior Mesenteric Artery Aneurysm with Arteriovenous Fistula: A Case Report.
Inferior mesenteric artery (IMA) aneurysm is a rare occurrence, accounting for 1% of all visceral artery aneurysms and is often found incidentally. Surgical resection and endovascular intervention have been first-line treatments because IMA aneurysms have a relatively high risk of life-threatening rupture. Herein, we report the case of a 57-year-old man having a large IMA aneurysm with an arteriovenous fistula that was treated conservatively. The IMA aneurysm was incidentally found using computed tomography (CT) and was connected to the splenic vein through the abnormally dilated tortuous vessels of an arteriovenous fistula. Surgical resection was planned initially; however, preoperative follow-up CT revealed that the aneurysm had shrunk with the growth of an intraluminal thrombus. Subsequently, the condition was conservatively managed with serial CT follow-up. Two years after the first visit, the aneurysm had shrunk and been completely replaced with a thrombus.


Claudin-1, -2, -3, -4, -5, and -7 in usual interstitial pneumonia and sarcoidosis.
The aim of this study was to study the expression of various claudins in sarcoidosis, usual interstitial pneumonia (UIP), and normal human lung. The expression and cell-specific localization of claudin-1, -2, -3, -4, -5, and -7 was analyzed by IHC. Bronchiolar epithelial cells showed mostly strong expression for claudin-1, -2, -3, -4, and -7 and mainly weak expression for claudin-5 in UIP, sarcoidosis, and normal lung. Three claudins, claudin-3, -4, and -7, were expressed in normal alveolar epithelium, mainly in type II pneumocytes. Claudin-5 was expressed strongly in endothelium of normal lung, and its staining was extremely intense in endothelium of UIP. Moderate or strong expression for claudin-1, -2, -3, -4, and -7 was observed in metaplastic alveolar- and bronchiolar-type epithelium in UIP and also in metaplastic alveolar-type epithelium in sarcoidosis. Expression of claudin-5 was mainly weak in metaplastic alveolar- and bronchiolar-type epithelium in UIP. We conclude that claudin-1, -2, -3, -4, -5, and -7 are expressed in UIP and sarcoidosis, and furthermore, the most prominent enhancement of staining is localized in metaplastic alveolar- and bronchiolar-type epithelium in UIP compared with the healthy lung.


Overview of age-related changes in psychomotor and cognitive functions in a prosimian primate, the gray mouse lemur (Microcebus murinus): Recent advances in risk factors and antiaging interventions.
Aging is not homogeneous in humans and the determinants leading to differences between subjects are not fully understood. Impaired glucose homeostasis is a major risk factor for cognitive decline in middle-aged humans, pointing at the existence of early markers of unhealthy aging. The gray mouse lemur (Microcebus murinus), a small lemuriform Malagasy primate, shows relatively slow aging with decreased psychomotor capacities at middle-age (around 5-year old). In some cases (∼10%), it spontaneously leads to pathological aging. In this case, some age-related deficits, such as severe cognitive decline, brain atrophy, amyloidosis, and glucoregulatory imbalance are congruent with what is observed in humans. In the present review, we inventory the changes occurring in psychomotor and cognitive functions during healthy and pathological aging in mouse lemur. It includes a summary of the cerebral, metabolic, and cellular alterations that occur during aging and their relation to cognitive decline. As nutrition is one of the major nonpharmacological antiaging strategies with major potential effects on cognitive performances, we also discuss its role in brain functions and cognitive decline in this species. We show that the overall approach of aging studies in the gray mouse lemur offers promising ways of investigation for understanding, prevention, and treatments of pathological aging in humans.


Vascular endothelial cell growth factor and fibroblast growth factor 2 expression in patients with critical limb ischemia.
For patients with critical limb ischemia (CLI), there is a great need for alternative treatment strategies. One option is therapeutic angiogenesis by administration of vascular growth factors. The lack of convincing clinical data supporting this strategy may be due to the ignorance of endogenous angiogenic processes in CLI. To evaluate the importance of vascular growth factors in the pathogenesis in CLI and provide information for clinical growth factor treatment trials, we determined the levels of vascular endothelial growth factor (VEGF) and fibroblast growth factor 2 (FGF-2) in the ischemic legs of patients with this disease. Skin and muscle biopsies from the calf and groin were gathered from 25 patients with CLI. Control samples came from 10 orthopedic patients and from 5 patients who were undergoing coronary artery bypass. The concentration of VEGF and FGF-2 in the biopsies was measured by enzyme-linked immunoassay, and to localize growth factor production, biopsied sections were immunostained. Patients with CLI had lower levels of VEGF in distal skin samples than in proximal ones (mean difference: 16.7 pg/mg total protein, 95% confidence interval: -1.0 to -32.3, P =.038), but these levels were similar to those in distal samples from control subjects (8.0, -4.6 to 20.5, P =.65). In muscle, VEGF concentrations were similar in calf and groin (5.4, -12.4 to 23.1, P =.55), but distal levels were higher than in distal samples from control subjects (23.7, 1.2 to 56.7, P =.028). Skin FGF levels tended to be higher in distal samples (45.3, 26.5 to 117.5, P =.090) and were higher than in skin from control subjects (106.2, -11.4 to 223.8, P =.050). Also in muscle, distal samples had higher levels of FGF-2 (35.6, -1.6 to 59.7, P =.006), but these levels were similar to what was found in control subjects (29.4., -16.3 to 81.2, P =.39). Growth factors were located in connective tissue between muscle fibers. In skin, the predominant FGF-2 staining was just below the epidermal layer, whereas VEGF appeared in the dermal layer. The results indicate that there are elevated concentrations of FGF-2 in calf muscle, whereas VEGF concentrations do not appear to be higher in the ischemic part of the leg in patients with CLI. These findings suggest that VEGF supplementation may be a more appropriate strategy for therapeutic angiogenesis to the calf area for CLI than FGF-2.


Surgical Repair of Mid-shaft Hypospadias Using a Transverse Preputial Island Flap and Pedicled Dartos Flap Around Urethral Orifice.
To evaluate the effects, particularly the incidence of anastomotic fistula, of a pedicled dartos flap around the urethral orifice in the treatment of urethroplasty of mid-shaft hypospadias. A total of 46 cases of congenital mid-shaft hypospadias were included in this study. The patients ranged in age from 0.7 to 25.4 years and the average was 5.8 years. The patients received penis chordee correction. A transverse preputial island flap was developed for urethral reconstruction. The proximal dartos of the urethral orifice was used to develop a pedicled dartos flap, which was transposed to cover and strengthen neourethral anastomosis. The ventral penile skin defect was repaired by another flap. The 46 patients were examined during follow-up visits for 6 months to 3 years. An anastomotic fistula was observed in one case (2.2 %). Scar healing without fistula was observed in another patient due to poor blood supply to part of the ventral penile skin. No other incidences of fistula, urethral rupture, flap necrosis, wound infections, urinary tract (meatal) stenosis, or urethral diverticulum were observed in the patients. A pedicled dartos flap around the urethral orifice can take advantage of well-vascularized local tissue to add a protective layer to the proximal aspect of the neourethral anastomosis for reducing the incidence of anastomotic fistula in mid-shaft hypospadias repair using a transverse preputial island flap. This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .


Increase in phospholamban content in mouse skeletal muscle after denervation.
It is well-known that denervation of motor nerves induces atrophy and decreases contractile force of the skeletal muscle. However, it is not completely understood how denervation alters calcium handling in the skeletal muscle. We investigated the effect of denervation on the expression and function of proteins involved in calcium handling. Two weeks after denervation of the right sciatic nerve in mice, we observed a significant decrease in mass and cross-sectional area of the ipsilateral tibialis anterior (TA) and flexor digitorum brevis (FDB) muscles. Also, we observed a significant decrease in the specific tetanus contractile force in the ipsilateral TA muscle. Calcium imaging of the ipsilateral FDB showed that the peak twitch and tetanus calcium concentrations were significantly decreased due to a decrease in calcium content of the sarcoplasmic reticulum (SR). Denervation reduced the maximum rate of sarcoplasmic/endoplasmic calcium ATPase (SERCA) activity. Interestingly, the amount of phospholamban (PLN), but not its transcripts, was increased in the ipsilateral vs. contralateral side after denervation, suggesting that denervation impairs constitutive regulation of PLN. Immunohistochemical analysis revealed increased PLN in all major fiber types in TA with IIx fibers showing a threefold higher expression than the contralateral side. These results suggest that the abnormal increase in PLN in the ipsilateral fast-twitch fibers may be involved in decreased SERCA activity, SR calcium content, peak calcium transients, and contractile forces of denervated muscles.


A tropical sediment toxicity test using the dipteran Chironomus crassiforceps to test metal bioavailability with sediment pH change in tropical acid-sulfate sediments.
The wetlands of the Magela floodplain of northern Australia, which is the major sink for dissolved metals transported in the Magela Creek system, contain acid-sulfate sediments. The rewetting of oxidized acid-sulfate soil each wet season produces acidic pulses that have the potential to alter the bioavailability of sediment-associated metal contaminants. Acute toxicity tests (72-h mean lethal concentration [LC50]) using the tropical chironomid Chironomus crassiforceps Kieffer showed that copper toxicity decreased from 0.64 mg/L at pH 6 to 2.30 mg/L at pH 4. Uranium toxicity showed a similar trend (36 mg/L at pH 6 and 58 mg/L at pH 4). Sediment toxicity tests developed using C. crassiforceps also showed that both metals were less toxic at the lower sediment pH with pore-water copper toxicity having a lowest-observed-effect concentration of 4.73 mg/L at pH 4 compared to 1.72 mg/L at pH 6. However, a lower pH increased pore-water metal concentrations and overlying water concentrations in bioassays. Hydrogen ion competition on metal receptor sites in C. crassiforceps was proposed to explain the decrease in toxicity in response to increased H+ activity. This study highlights the need to consider site-specific physicochemical conditions before applying generic risk assessment methods.


[Behaviour of ATP and lactate in human papillary muscle during profound hypothermia and injection cardioplegia with magnesium-asparatate-procaine (author's transl)].
In 53 patients with mitral- or aortic-mitral valve disease, the content of ATP and lactate of the papillary muscles resected at the time of valve replacement was investigated at the beginning of ischemic arrest and at the time of reperfusion. Profound body hypothermia (25 degrees C) and injection cardioplegia using magnesium-aspartate-procaine were applied for myocardial protection. In hypertrophic papillary muscles the myocardial ATP content decreased at a slower rate (ATP decay 12% of the initial value after 60 minutes of ischemia) than in normal papillary muscles obtained from patients with isolated mitral stenosis (ATP decay 33% of the initial value after 40 minutes of ischemia). 20% of the patients required temporary inotropic circulatory support postoperatively for 12 to 88 hours. The ATP content of the papillary muscles of these patients differed only little from those, in who no myocardial failure occurred. However the myocardial lactate levels were higher in patients in whom a low cardiac output state evolved.


Simple gas-liquid chromatographic method for the measurement of disopyramide in blood-plasma or serum and in urine.
A simple method has been developed for the measurement of disopyramide in blood-plasma or serum at the concentrations attained during therapy. A relatively small (200 microliter) sample volume is made basic and extracted with 50 microliter of chloroform containing an internal standard, and the extract is analysed directly by gas-liquid chromatography with flame-ionization detection. The instrument calibration is linear and passes through the origin of the graph. Neither solvent transfer nor evaporation steps are used in the extraction procedure, which takes less than 3 min to complete, and urine specimens may be analysed by an analogous technique. No interference from either endogenous sample constituents or other drugs has been observed, although a simple back-extraction procedure is described which eliminates potential interference from a small number of basic and neutral drugs.


Antimicrobial resistance and population structure of Staphylococcus epidermidis recovered from animals and humans.
While Staphylococcus epidermidis, as part of the commensal flora, is a well-known human opportunistic pathogen, only little is known about the genetic relatedness of S. epidermidis carriage isolates from animal and human origin. This study aimed to compare S. epidermidis recovered from livestock, livestock-farmers and humans associated with the hospital environment. A total of 193 S. epidermidis isolates from three populations [animals (n=33), farmers (n=86) and hospital-associated (n=74)] were characterized by broth microdilution antimicrobial susceptibility testing, staphylococcal cassette chromosome mec (SCCmec) typing, pulsed field gel electrophoresis (PFGE) and multilocus sequence typing (MLST). The overall S. epidermidis nasal colonization rate was low in animals (1-9%) but high among farmers (75%). High levels of multi-resistance were found in all populations. Tetracycline resistance was high in animal and farmer isolates; resistance to erythromycin, clindamycin and trimethoprim was high in animal and hospital-associated isolates. Methicillin-resistant S. epidermidis - MRSE isolates were found in all collections, with 22 (67%) MRSE in animals, 44 (51%) MRSE in farmers and 42 (57%) MRSE associated with the hospital-setting. Known SCCmec types and variants were detected in 79% of MRSE; the rest were non-typeable cassettes. In total 79 PFGE-types were found, of which 22 were shared between livestock, farmers and the hospital settings. Clonal complex 2 was predominant in all three populations and most STs corresponded to types previously observed in community and nosocomial S. epidermidis populations. S. epidermidis isolates from livestock, farmers and hospital-setting showed a high level of diversity, but some clones can be found in humans as well as in animals.


Cell kinetic analysis of murine squamous cell carcinomas: a comparison of single versus double labelling using flow cytometry and immunohistochemistry.
The study was originally set up to measure accurate cell kinetic parameters in two murine squamous cell carcinomas (scc) for comparison with radiobiological data on proliferation during radiotherapy. The tumours, AT84 and AT478, were both moderately well differentiated aneuploid scc. In the course of the study, several comparisons of techniques were made in two different centres. This paper reports on the results of those comparisons involving two different detection methods (flow cytometry and immunohistochemistry), single vs double labelling, and in vivo and in vitro labelling, the latter using tissue slices incubated under high pressure oxygen. Pulse labelling studies with bromodeoxyuridine (BrdUrd) showed that the labelling indices (LI) were not significantly different after in vitro or in vivo labelling. In addition, the flow cytometry (FCM) and immunohistochemistry (IHC) methods also gave labelling indices which were not significantly different. Only tumour cells were analysed in these studies by selecting cells on the basis of aneuploidy (FCM) or morphology (IHC). The DNA synthesis time of the tumour cells were analysed by both techniques. For FCM, the Relative Movement method was used (Begg et al., 1985). For IHC, a double labelling method was used, employing BrdUrd and triated thymidine (3H-TdR) administered several hours apart, detected simultaneously using immunoperoxidase and autoradiography, respectively. When both labels were administered in vivo, there was good agreement for Ts between the FCM and IHC methods. Attempts were also made to measure Ts in vitro using both techniques. With double labelling, it was found that cells did not take up the second label, implying a failure of cycle progression. This was confirmed by FCM results, showing no movement of labelled cells through the S-phase, despite an initially high uptake. This could not be influenced by lowering the DNA precursor concentration or by adding foetal calf serum. This indicates that DNA synthesis times are difficult or impossible to measure in vitro in fresh tumour explants. Finally, the double labelling IHC method allowed intratumoural variations of both LI and Ts to be studied. Both parameters were found to vary markedly throughout the tumour volume, particularly for larger tumours (600 mg), giving calculated local potential doubling time values (Tpot) ranging from 1-7 days.


Long-term outcomes of endoscopic gallbladder stenting in high-risk surgical patients with calculous cholecystitis (with videos).
Recently, endoscopic gallbladder stenting (EGBS) has been performed to prevent recurrences in high-risk surgical patients with cholecystitis. However, evidence regarding the long-term outcomes of EGBS is sparse. We investigated the cholecystitis recurrence rate in high-risk surgical patients with acute calculous cholecystitis and compared the cholecystitis recurrence rates in patients in whom EGBS was performed with those in patients who were observed after percutaneous drainage. We studied 64 consecutive high-risk surgical patients with acute calculous cholecystitis who required gallbladder decompression between 2007 and 2014. We divided the patient cohort into patients who underwent observation after percutaneous drainage between 2007 and 2011 (OAPD group) and those who underwent EGBS between 2012 and 2014 (EGBS group), and we compared the groups. The technical success rate of EGBS was 82.9% based on the intention-to-treat analysis. The cholecystitis recurrence rates were 17.2% in the OAPD group and 0% in the EGBS group, a difference that was significant (P = .043). There was also a significant difference between the groups with respect to the time to recurrent cholecystitis, which was determined by using Kaplan-Meier analysis (P = .015). The overall biliary event rates were 24.1% in the OAPD group and 9.1% in the EGBS group, and no significant difference was noted (P = .207). EGBS reduced the recurrence of cholecystitis in high-risk surgical patients with calculous cholecystitis. However, stent-related adverse events may occur, and modifications are necessary to reduce these.


Enhancing full-length antibody production by signal peptide engineering.
Protein secretion to the periplasm of Escherichia coli offers an attractive route for producing heterologous proteins including antibodies. In this approach, a signal peptide is fused to the N-terminus of the heterologous protein. The signal peptide mediates translocation of the heterologous protein from the cytoplasm to the periplasm and is cleaved during the translocation process. It was previously shown that optimization of the translation initiation region (TIR) which overlaps with the nucleotide sequence of the signal sequence improves the production of heterologous proteins. Despite the progress, there is still room to improve yields using secretion as a means to produce protein complexes such as full-length monoclonal antibodies (mAbs). In this study we identified the inefficient secretion of heavy chain as the limitation for full-length mAb accumulation in the periplasm. To improve heavy chain secretion we investigated the effects of various signal peptides at controlled TIR strengths. The signal peptide of disulfide oxidoreductase (DsbA) mediated more efficient secretion of heavy chain than the other signal peptides tested. Mutagenesis studies demonstrated that at controlled translational levels, hydrophobicity of the hydrophobic core (H-region) of the signal peptide is a critical factor for heavy chain secretion and full-length mAb accumulation in the periplasm. Increasing the hydrophobicity of a signal peptide enhanced heavy chain secretion and periplasmic levels of assembled full-length mAbs, while decreasing the hydrophobicity had the opposite effect. This study demonstrates that under similar translational strengths, the hydrophobicity of the signal peptide plays an important role in heavy chain secretion. Increasing the hydrophobicity of the H-region and controlling TIR strengths can serve as an approach to improve heavy chain secretion and full-length mAb production in E. coli.


Age related changes in the lipids of the microsomal and the mitochondrial membranes of rat liver and kidney.
The lipid contents of the microsomal and mitochondrial membrane fractions of liver and kidney were determined in 6 and 24 month old rats. A significant age related increase in the molar ratio of cholesterol/phospholipid was observed in all membrane fractions. A significant age related reduction of phospholipid was noted in the microsomal fractions of liver and kidney. The relative amount of phosphatidylethanolamine was found to decrease in all membrane fractions during aging. Membrane glyceride content, however, remained relatively constant with age. Significant increase in oleic acid was seen in the neutral lipid of both liver and kidney and in the polar lipid of kidney. Significant increase in docosahexaenoic acid and significant decrease in linoleic acid were seen in the polar lipid of the liver membrane fractions. Possible alterations in membrane physiochemical properties and in membrane function due to these age related lipid changes are discussed.


Induction of inducible nitric oxide synthase expression in ammonia-exposed cultured astrocytes is coupled to increased arginine transport by upregulated y(+)LAT2 transporter.
One of the aspects of ammonia toxicity to brain cells is increased production of nitric oxide (NO) by NO synthases (NOSs). Previously we showed that ammonia increases arginine (Arg) uptake in cultured rat cortical astrocytes specifically via y(+)L amino acid transport system, by activation of its member, a heteromeric y(+)LAT2 transporter. Here, we tested the hypothesis that up-regulation of y(+)LAT2 underlies ammonia-dependent increase of NO production via inducible NOS (iNOS) induction, and protein nitration. Treatment of rat cortical astrocytes for 48 with 5 mM ammonium chloride ('ammonia') (i) increased the y(+)L-mediated Arg uptake, (ii) raised the expression of iNOS and endothelial NOS (eNOS), (iii) stimulated NO production, as manifested by increased nitrite+nitrate (Griess) and/or nitrite alone (chemiluminescence), and consequently, (iv) evoked nitration of tyrosine residues of proteins in astrocytes. Except for the increase of eNOS, all the above described effects of ammonia were abrogated by pre-treatment of astrocytes with either siRNA silencing of the Slc7a6 gene coding for y(+)LAT2 protein, or antibody to y(+)LAT2, indicating their strict coupling to y(+)LAT2 activity. Moreover, induction of y(+)LAT2 expression by ammonia was sensitive to Nf-κB inhibitor, BAY 11-7085, linking y(+)LAT2 upregulation to the Nf-κB activation in this experimental setting as reported earlier and here confirmed. Importantly, ammonia did not affect y(+)LAT2 expression nor y(+)L-mediated Arg uptake activity in the cultured cerebellar neurons, suggesting astroglia-specificity of the above described mechanism. The described coupling of up-regulation of y(+)LAT2 transporter with iNOS in ammonia-exposed astrocytes may be considered as a mechanism to ensure NO supply for protein nitration. Ammonia (NH4(+)) increases the expression and activity of the L-arginine (Arg) transporter (Arg/neutral amino acids [NAA] exchanger) y(+)LAT2 in cultured rat cortical astrocytes by a mechanism involving activation (nuclear translocation) of the transcription factor nuclear factor-Nuclear factor-κB (Nf-κB-p65). Up-regulation of y(+)LAT2 transporter is coupled with increased inducible nitric oxide synthase (iNOS) expression, which leads to increase nitric oxide (NO) synthesis and protein nitration.


Antibacterial effect of nanocurcumin inside the implant fixture: An in vitro study.
Infections after implant placement are the main reasons for the failure of implant treatments. The present study aimed to evaluate the antibacterial effects of nanocurcumin inside the implant fixture against Escherichia coli, Staphylococcus aureus, and Enterococcus faecalis. Twenty seven implants were classified in three groups for testing the antibacterial effect of nanocurcumin, chlorhexidine (as negative control), and distilled water (as negative control). Each group was then divided into three subgroups to study the effect of the applied torque on the antimicrobial effect of nanocurcumin. All implant abutment assemblies were submerged in bacteria suspension and were incubated at 37°C for 24 hours. The contents of each implant were removed to count the colony of bacteria on the surface of plates containing nutrient agar. Results indicated that the inhibitory rate of bacteria by nanocurcumin was above 99% in all bacteria. Besides, by increasing the amount of applied torque from 10 to 35 N.cm, the CFU of bacteria in exposure to nanocurcumin significantly were decreased (p-value < 0.01). The results of this study revealed that nanocurcumin can be used inside the implant fixture in order to use antimicrobial effects and further stabilization and success of the implant.


Prevalence and outcome of methicillin-resistant Staphylococcus aureus colonization in two nursing centers in Georgia.
To evaluate the cost-effectiveness and validity of aggressive diagnosing and eradication of methicillin-resistant Staphylococcus aureus (MRSA) colonization. An observational study of MRSA colonization. Two skilled nursing facilities in Georgia. Eighty-seven residents of the above-mentioned skilled nursing facilities No changes were made in either nursing facility practices or patient treatments. The presence or absence of MRSA colonization was determined at the beginning of the study, after hospitalizations or absences from the facility, and 12 months after initial testing. Eighty-seven participants began the study, and 56 completed it. At the beginning of the study, 17.9% of the 56 who later completed the study were positive for MRSA. By the end of the study, colonization had decreased to 14.3%. The difference in the number of positive cultures for residents returning from the hospital compared with those returning from non-hospital settings was not statistically significant (P = .546). No deaths or hospitalizations were attributable to MRSA infection. Because no significant correlation was found between MRSA colonization and active MRSA-related infections, patients colonized by MRSA should not be barred from admission to nursing homes.


Arginase inhibitor attenuates pulmonary artery hypertension induced by hypoxia.
Hypoxia-induced pulmonary arterial hypertension (HPAH) is a refractory disease characterized by increased proliferation of pulmonary vascular smooth cells and progressive pulmonary vascular remodeling. The level of nitric oxide (NO), a potential therapeutic vasodilator, is low in PAH patients. L-arginine can be converted to either beneficial NO by nitric oxide synthases or to harmful urea by arginase. In the present study, we aimed to investigate whether an arginase inhibitor, S-(2-boronoethyl)-L-cysteine ameliorates HPAH in vivo and vitro. In a HPAH mouse model, we assessed right ventricle systolic pressure (RVSP) by an invasive method, and found that RSVP was elevated under hypoxia, but was attenuated upon arginase inhibition. Human pulmonary artery smooth muscle cells (HPASMCs) were cultured under hypoxic conditions, and their proliferative capacity was determined by cell counting and flow cytometry. The levels of cyclin D1, p27, p-Akt, and p-ERK were detected by RT-PCR or Western blot analysis. Compared to hypoxia group, arginase inhibitor inhibited HPASMCs proliferation and reduced the levels of cyclin D1, p-Akt, p-ERK, while increasing p27 level. Moreover, in mouse models, compared to control group, hypoxia increased cyclin D1 expression but reduced p27 expression, while arginase inhibitor reversed the effects of hypoxia. Taken together, these results suggest that arginase plays an important role in increased proliferation of HPASMCs induced by hypoxia and it is a potential therapeutic target for the treatment of pulmonary hypertensive disorders.


Truncal duplication: a case report.
Truncal duplication represents one of the rare forms of conjoined twins. We observed a male infant with a truncal duplication; in the host twin no pathology was found except an atrial septal defect and a large omphalocele. The parasitic twin was attached to the xiphoid region (xiphopagus) in an opposite and "horseriding" manner, was acephalic, and had multiple gastrointestinal, genitourinary, and skeletal anomalies. The junction site consisted of lipoid and muscular structures. Surgical separation was done without any difficulty. The abdominal defect was repaired primarily. The host twin is doing well.


Dusp6 deficiency attenuates neutrophil-mediated cardiac damage in the acute inflammatory phase of myocardial infarction.
Dual-specificity phosphatase 6 (DUSP6) serves a specific and conserved function on the dephosphorylation of extracellular signal-regulated kinase 1/2 (ERK1/2). We previously identified Dusp6 as a regenerative repressor during zebrafish heart regeneration, therefore we propose to investigate the role of this repressor in mammalian cardiac repair. Utilizing a rat strain harboring Dusp6 nonsense mutation, rat neutrophil-cardiomyocyte co-culture, bone marrow transplanted rats and neutrophil-specific Dusp6 knockout mice, we find that Dusp6 deficiency improves cardiac outcomes by predominantly attenuating neutrophil-mediated myocardial damage in acute inflammatory phase after myocardial infarction. Mechanistically, Dusp6 is transcriptionally activated by p38-C/EBPβ signaling and acts as an effector for maintaining p-p38 activity by down-regulating pERK and p38-targeting phosphatases DUSP1/DUSP16. Our findings provide robust animal models and novel insights for neutrophil-mediated cardiac damage and demonstrate the potential of DUSP6 as a therapeutic target for post-MI cardiac remodeling and other relevant inflammatory diseases.


Determining efficacy of monitoring devices on ceramic bond to resin composite.
This paper aims to assess the effectiveness of 3D nanoroughness and 2D microroughness evaluations, by their correlation with contact angle measurements and shear bond strength test, in order to evaluate the effect of two different acids conditioning on the bonding efficacy of a leucite-based glass-ceramic to a composite resin. Ceramic (IPS Empress) blocks were treated as follows: 1) no treatment, 2) 37% phosphoric acid (H3PO4), 15 s, 3) 9% hydrofluoric acid (HF), 5 min. Micro- and nano-roughness were assessed with a profilometer and by means of an atomic force microscopy (AFM). Water contact angle (CA) measurements were determined to assess wettability of the ceramic surfaces with the asixymetric drop shape analysis contact diameter technique. Shear bond strength (SBS) was tested to a resin composite (Z100) with three different adhesive systems (Scotchbond Multipurpose Plus, Clearfil New Bond, ProBOND). Scanning electron microscopy (SEM) images were performed. nanoroughness values assesed in 50x50 µm areas showed differences between groups did not found by profilometer. HF treatment created the nano- roughest surfaces and the smallest CA (p<0.05), producing the highest SBS to the composite resin with all tested adhesive systems (p<0.05). No differences existed between the SBS produced by the adhesive systems evaluated with any of the surface treatments tested. Nano-roughness obtained in a 50x50 µm scan size areas was the most reliable data to evaluate the topographical changes produced by the different acid treatments on ceramic surfaces.


Studies of water permeability and proteins of erythrocyte membranes in patients with Duchenne muscular dystrophy.
The characteristics of water permeability of erythrocytes from 54 Duchenne muscular dystrophy (DMD) patients and age-matched controls have been determined by a pulse nuclear magnetic resonance (NMR) technique. A decreased permeability of erythrocyte membrane in DMD was definitely found at all temperatures between 15 and 42 degrees C, with normal values for the activation energy of water diffusion. No differences between DMD and control subjects in the pattern of erythrocyte membrane polypeptides separated by two-dimensional electrophoresis could be detected. The findings are discussed in relation to the molecular mechanisms of water diffusion across erythrocyte membrane and the problem of erythrocyte membrane abnormalities in DMD. A new interpretation of erythrocyte membrane alterations is proposed based on the recent findings regarding the molecular pathology of DMD.


Analyses of a Bacillus subtilis homologue of the Na+/H+ antiporter gene which is important for pH homeostasis of alkaliphilic Bacillus sp. C-125.
Bacillus subtilis was revealed to have a homologous region to the DNA fragment responsible for alkaliphily of alkaliphilic Bacillus sp. C-125 on the genome, as reported previously [1]. The yufT gene on the B. subtilis genome showed a significant similarity with ORF1 of Bacillus sp. C-125, which is related to membrane potential (DeltaPsi)-driven Na+/H+ antiport activity and is important for pH homeostasis in an alkaline condition. Disruption of the yufT gene resulted in the decrease of Na+/H+ antiport activity, and the growth of the yufT disrupted strain was impaired with an increase in the external Na+ concentration. We conclude that the yufT gene encodes a Na+/H+ antiporter, which has a dominant role in the extrusion of cytotoxic Na+.


Affective impairment in young acute schizophrenics: its structure, course and prognostic significance.
In order to investigate the structure, longitudinal course, and significance of affect impairments in young acute schizophrenics, we prospectively studied a group of 37 consecutively admitted acute inpatients at baseline and 26 months later (n = 19). They were evaluated at both points on a multidimensional affect rating scale, psychopathology rating scales, and tests of attention and motorium. Additionally, they were assessed on historical and subdiagnostic variables at baseline and on level of functioning at follow-up. Factor analysis revealed three reliably measured affect components: emotional unrelatedness, expressive immobility, and inappropriateness of affect. The first two factors were strongly interrelated and associated also with the BPRS depression factor at baseline. All four affective dimensions proved unstable over the course of 2 years and produced different sets of correlates at the two points of time. In the acute phase, only the depression factor carried favorable prognostic import (r = 0.52, P less than 0.05). Emotional unrelatedness and expressive immobility were of unfavorable consequence only when observed in the follow-up stage. Thus, the affect profile seemed to be an evolving rather than a static phenomenon, precluding generalizations across the early course of schizophrenia. Possible interpretations of the baseline overlap between affect deficits and depression were discussed as well as their different prognostic implications.


Intraoperative and postoperative anaesthetic and analgesic effect of multipoint transcutaneous electrical acupuncture stimulation combined with sufentanil anaesthesia in patients undergoing supratentorial craniotomy.
To investigate the anaesthetic and analgesic effect of multipoint transcutaneous electrical acupuncture stimulation (TEAS) during supratentorial tumour resection for postoperative recovery and side effects. In a blinded clinical trial, 92 patients scheduled for supratentorial craniotomy under general anaesthesia were randomly allocated into either a multipoint TEAS (n=46) or a sham TEAS group (n=46). All patients received total intravenous anaesthesia (TIVA) with propofol and sufentanil. The target concentration of sufentanil was adjusted and recorded according to mean arterial pressure (MAP), heart rate (HR) and bispectral index (BIS). Patients in the TEAS group received TEAS 30 min before anaesthesia induction and this was maintained throughout the operation at four pairs of acupuncture points. Postoperative pain, recovery and side effects were evaluated. Eighty-eight patients completed the study. Continuous monitoring of MAP, HR and BIS showed stable values with no significant differences between the two groups (p>0.05). Sufentanil target plasma concentration in TEAS patients was significantly lower at some time points during supratentorial craniotomy, and total sufentanil consumption was significantly higher in the sham group (p<0.05). Postoperative recovery and pain were significantly improved by TEAS (p<0.001), without the postoperative side effects. Multipoint TEAS at both proximal and distal points combined with TIVA can significantly decrease intraoperative sufentanil requirements, increase pain relief on postoperative day 1 and improve postoperative recovery of patients during supratentorial tumour resection, with no significant increase of side effects. These findings suggest that multipoint TEAS may be clinically effective as an adjunct to analgesia in intraoperative anaesthesia and postoperative pain treatment and may speed recovery. Chinese Clinical Trial Registry (registration number ChiCTR-TRC-10001078).


Effects and acceptance of semipermeable gloves compared to cotton gloves in patients with hand dermatoses: Results of a controlled intervention study.
Semipermeable gloves might be an alternative to cotton gloves in management of hand dermatoses. To compare acceptance and tolerability of gloves made of cotton or a semipermeable Sympatex membrane in patients with hand dermatoses and their effect on skin lesions when worn overnight. A total of 199 patients with work-related symmetrical hand dermatoses were asked to wear a cotton glove (COT) on one hand and a Sympatex glove underneath a cotton glove (SYM/COT) on the other hand for 19 consecutive nights. The severity of skin lesions was regularly examined. A questionnaire on acceptance and tolerability was used in a subset of 126 patients. Data on skin lesions were available for 183 participating patients (92%). The severity did not differ substantially between hands covered with SYM/COT or COT over time. Questionnaire data were available for 120 patients (95%). SYM/COT received better ratings regarding climate conditions while COT showed superiority in wearing comfort, practicality and appearance. Both SYM/COT and COT were well tolerated and accepted in patients with hand dermatoses. Hence, SYM/COT may serve as suitable alternative for COT as comfort gloves supporting therapeutic efforts. The observed slight differences may influence the decision when selecting the glove type.


Recent developments of small molecules with anti-inflammatory activities for the treatment of acute lung injury.
Acute Lung Injury (ALI) can be caused by various diseases or conditions such as sepsis, pneumonia, trauma, shock, and inhalation of toxic gases. Many efforts have been made to identify new agents capable of treating ALI, and many compounds have shown interesting activities in the treatment of ALI. However, most of these compounds have only been tested for their clinical significance using in vitro and in vivo animal models. In this review, the developments in the design and structural modification of effective active agents for the treatment of ALI is summarized. Firstly, the application of α, β-unsaturated carbonyl moieties as an importantly superior framework in the development of natural product-derived anti-ALI agents is described. As well, the biological activities of the hybrid derivatives from natural products are discussed. Secondly, the potential of synthetic small molecule active compounds in the treatment of ALI is demonstrated. In addition, the structure activity relationship (SAR) and possible mechanisms of action in new chemical molecular entities were investigated. This present mini-review will be useful to scientists in research fields of medicinal chemistry, organic synthesis, and also various biological applications particularly for the development of novel anti-ALI agents.


Natural anti-chondrocyte cytotoxic activity in the mouse: supportive role of macrophages.
The role of macrophages (M phi) in natural anti-chondrocyte cytotoxic activity of normal murine splenocytes (SPL) and peritoneal cells (PC) was examined by means of an 18-hr 51Cr-release assay. Removal of M phi by either plastic adherence or carbonyl iron and magnet resulted in significant (P less than 0.01) reduction of SPL-mediated lysis of chondrocytes. It, however, did not influence natural anti-YAC-1 leukemia activity. On the contrary, M phi-depleted suspensions of PC retained their anti-chondrocyte activity at the initial level. Neither adherent SPL nor adherent PC exerted significant anti-chondrocyte cytotoxicity. The activity of nonadherent SPL could be, however, restored by reconstitution with the graded numbers of syngeneic adherent spleen- or peritoneal cavity-derived cells. Restoration of anti-chondrocyte activity of nonadherent SPL to the level comparable with that of untreated SPL was achieved with about 10% of adherent cells. On the other hand, reconstitution of nonadherent PC did not result in an increase of chondrocyte lysis. Potentiation of nonadherent SPL activity was seen only with syngeneic M phi, and the latter could not be replaced by conditioned media which were generated in the 18-hr culture of adherent SPL or PC.


Interaction among 5,10 methylenetetrahydrofolate reductase, plasminogen activator inhibitor and endothelial nitric oxide synthase gene polymorphisms predicts the severity of coronary artery disease in Turkish patients.
Genetic factors play a role in the onset of coronary artery disease. The objective of our study is to evaluate the single locus and combined effects of three different genetic polymorphisms (methylenetetrahydrofolate reductase C677T polymorphism, plasminogen activator inhibitor 4G/5G polymorphism, and endothelial nitric oxide synthase 3-27 base pairs repeat polymorphism) on the presence and extent of coronary artery disease in patients with early-onset coronary artery disease. DNA samples were obtained from 102 consecutive patients with symptoms resulting from early-onset coronary artery disease documented by coronary angiography. The severity of coronary artery disease in patients was stratified into three groups as one, two, or three-vessel coronary artery disease. The control group was selected from older subjects with a recent and negative cardiac work-up. Information on standard risk factors was collected. Multifactor dimensionality reduction analysis was performed to seek a model of coronary artery disease based on these three genetic polymorphisms. Single-locus effects of the three polymorphisms were not significantly related to the presence or severity of coronary artery disease. When gene-gene interactions were studied, however, the severity of disease was related to the frequency of high-risk alleles, yet advanced analysis did not detect a significant genetic model for coronary artery disease in these patients based on these three genetic polymorphisms. These three genetic polymorphisms are susceptibility loci and genotypes of these genes are neither necessary nor sufficient for the coronary artery disease to occur, but coexistence of high-risk alleles may increase the severity of coronary artery disease.


Sorting permutations by cut-circularize-linearize-and-paste operations.
Genome rearrangements are studied on the basis of genome-wide analysis of gene orders and important in the evolution of species. In the last two decades, a variety of rearrangement operations, such as reversals, transpositions, block-interchanges, translocations, fusions and fissions, have been proposed to evaluate the differences between gene orders in two or more genomes. Usually, the computational studies of genome rearrangements are formulated as problems of sorting permutations by rearrangement operations. In this article, we study a sorting problem by cut-circularize-linearize-and-paste (CCLP) operations, which aims to find a minimum number of CCLP operations to sort a signed permutation representing a chromosome. The CCLP is a genome rearrangement operation that cuts a segment out of a chromosome, circularizes the segment into a temporary circle, linearizes the temporary circle as a linear segment, and possibly inverts the linearized segment and pastes it into the remaining chromosome. The CCLP operation can model many well-known rearrangements, such as reversals, transpositions and block-interchanges, and others not reported in the biological literature. In addition, it really occurs in the immune response of higher animals. To distinguish those CCLP operations from the reversal, we call them as non-reversal CCLP operations. In this study, we use permutation groups in algebra to design an O(δn) time algorithm for solving the weighted sorting problem by CCLP operations when the weight ratio between reversals and non-reversal CCLP operations is 1:2, where n is the number of genes in the given chromosome and δ is the number of needed CCLP operations. The algorithm we propose in this study is very simple so that it can be easily implemented with 1-dimensional arrays and useful in the studies of phylogenetic tree reconstruction and human immune response to tumors.


Alveolar hemorrhage associated with a membranoproliferative glomerulonephritis and smooth muscle antibody.
Alveolar hemorrhage along with glomerulonephritis developed in a 31-year-old black man. Renal biopsy demonstrated a granular pattern of immunofluorescence along basement membrane glomeruli, suggesting an immunologically mediated illness; however, light microscopy revealed a pure membranoproliferative glomerulonephritis, which has not been described to date in association with the alveolar hemorrhage syndromes. The patient's immunologic profile revealed a positive antinuclear antibody titer along with the presence of smooth muscle antibodies.


Improving the quality of risk assessments in Canada using a principle-based approach.
In 2000, the Government of Canada issued the "Framework for Science and Technology Advice" (FSTA). This official government policy directive mandates that decision-making to protect human health and the environment should be carried out in a timely, predictable, and efficient manner. It also leaves no doubt that quality assurance and quality control (QA/QC) are required for both technical data and the decision-making process itself. Since current publicly available Canadian risk assessment guidance does not directly address quality management for the decision-making process, an evaluative scheme for CEPA risk assessments was created. For each of the six FSTA principles (early issue identification, inclusiveness, sound science and science advice, uncertainty and risk, openness and transparency, review) specific expectations were developed for performance measurement purposes. This scheme addresses the FSTA requirement for a policy compliance checklist. It could serve as the basis for quality assurance audits during risk assessment production and quality control audits in subsequent review of the risk assessment process. Among other things, it could also improve the integration of science advice, policy objectives and general risk communication by enhancing decision-making openness and transparency.


Glomerular Endothelial Cell-Derived microRNA-192 Regulates Nephronectin Expression in Idiopathic Membranous Glomerulonephritis.
Autoantibodies binding to podocyte antigens cause idiopathic membranous glomerulonephritis (iMGN). However, it remains elusive how autoantibodies reach the subepithelial space because the glomerular filtration barrier (GFB) is size selective and almost impermeable for antibodies. Kidney biopsies from patients with iMGN, cell culture, zebrafish, and mouse models were used to investigate the role of nephronectin (NPNT) regulating microRNAs (miRs) for the GFB. Glomerular endothelial cell (GEC)-derived miR-192-5p and podocyte-derived miR-378a-3p are upregulated in urine and glomeruli of patients with iMGN, whereas glomerular NPNT is reduced. Overexpression of miR-192-5p and morpholino-mediated npnt knockdown induced edema, proteinuria, and podocyte effacement similar to podocyte-derived miR-378a-3p in zebrafish. Structural changes of the glomerular basement membrane (GBM) with increased lucidity, splitting, and lamellation, especially of the lamina rara interna, similar to ultrastructural findings seen in advanced stages of iMGN, were found. IgG-size nanoparticles accumulated in lucidity areas of the lamina rara interna and lamina densa of the GBM in npnt-knockdown zebrafish models. Loss of slit diaphragm proteins and severe structural impairment of the GBM were further confirmed in podocyte-specific Npnt knockout mice. GECs downregulate podocyte NPNT by transfer of miR-192-5p-containing exosomes in a paracrine manner. Podocyte NPNT is important for proper glomerular filter function and GBM structure and is regulated by GEC-derived miR-192-5p and podocyte-derived miR-378a-3p. We hypothesize that loss of NPNT in the GBM is an important part of the initial pathophysiology of iMGN and enables autoantigenicity of podocyte antigens and subepithelial immune complex deposition in iMGN.


Tailored lamellar keratoplasty: a new technique.
We present a new method for treating complicated corneal gaps among patients who have previously undergone radial keratotomy (RK).After complete healing of a fungal keratitis in a patient who has undergone RK, we removed the oedematous corneal sector and put anterior tailored segment of a corneal button previously used for Descemet stripping endothelial keratoplasty. The patient's best-corrected visual acuity reached to 20/20 6 months later. This technique will compensate for oedematous parts without putting a significant effect on other parts such as purse string sutures.


Evaluation of platelet tests for measurement of cell integrity.
Various tests were evaluated for their capacity to differentiate between platelet suspensions with different degrees of cell damage. Those suspensions were prepared by simultaneous isolation of platelets from the same platelet-rich plasma (PRP) using the following procedures: 1. centrifugation at 4 degrees C with EDTA 2. gel filtration in Tangen's buffer 3. gel filtration in Ca2+-free Tyrode's soltuion 4. gel filtration in Ca2+-free Tyrode followed by dehydration against polyethylene glycol 20,000 and 5. albumin density gradient centrifugation. In these suspensions and in the original PRP the following parameters were studied: 1. morphology; 2. aggregability upon ADP addition; 3. platelet factor 3 availability; 4. uptake of 14C-serotonin and 3H-adenine; 5. metabolism of 3H-adenine and adenylate energy charge; 6. endogenous total ATP, ADP and serotonin and 7. lactate dehydrogenase (LDH) activity. Quantitation of pseudopod formation in the light or electron microscope and log dose response studies for ADP-induced aggregation proved to be the most sensitive and reproducible of the tests studied. Additional information could be obtained from measurement of the 3H-label in the ATP and hypoxanthine-inosine fractions and calculation of the adenylate energy charge. Determination of platelet factor 3 availability or uptake studies of 14C-serotonin and 3H-adenine were less suitable for discriminating between cell suspensions. Data for total ATP and serotonin concentrations and LDH activity differed between the cell suspensions but instead of detecting various degrees of cell damage they reflected alterations in platelet population caused by the isolation procedures.


Investigational Assay for Haplotype Phasing of the Huntingtin Gene.
Novel treatments for Huntington's disease (HD), a progressive neurodegenerative disorder, include selective targeting of the mutant allele of the huntingtin gene (mHTT) carrying the abnormally expanded disease-causing cytosine-adenine-guanine (CAG) repeat. WVE-120101 and WVE-120102 are investigational stereopure antisense oligonucleotides that enable selective suppression of mHTT by targeting single-nucleotide polymorphisms (SNPs) that are in haplotype phase with the CAG repeat expansion. Recently developed long-read sequencing technologies can capture CAG expansions and distant SNPs of interest and potentially facilitate haplotype-based identification of patients for clinical trials of oligonucleotide therapies. However, improved methods are needed to phase SNPs with CAG repeat expansions directly and reliably without need for familial genotype/haplotype data. Our haplotype phasing method uses single-molecule real-time sequencing and a custom algorithm to determine with confidence bases at SNPs on mutant alleles, even without familial data. Herein, we summarize this methodology and validate the approach using patient-derived samples with known phasing results. Comparison of experimentally measured CAG repeat lengths, heterozygosity, and phasing with previously determined results showed improved performance. Our methodology enables the haplotype phasing of SNPs of interest and the disease-causing, expanded CAG repeat of the huntingtin gene, enabling accurate identification of patients with HD eligible for allele-selective clinical studies.


An ascending spinal pathway transmitting a central rhythmic pattern to the magnocellular red nucleus in the cat.
The activity of cells in the magnocellular red nucleus (RNm) was recorded extra- and intracellularly in the curarized thalamic cat performing fictive locomotion. The locomotor episodes were detected from the rhythmic activity recorded in the motor nerves of the contralateral hindlimb. It was confirmed that, during fictive locomotion, a large proportion of the rubrospinal cells (56% in our sample) exhibit a rhythmic pattern of activity which is synchronized with the efferent spinal motor nerve activity. On the basis of the intracellular recordings it was established that phases of intense synaptic activity with mixed excitatory postsynaptic potentials (EPSPs) and inhibitory postsynaptic potentials (IPSPs) are involved in this rhythmicity. After eliminating the cerebellar input to the RNm, it was observed that the cells still received intense excitatory and inhibitory inputs, resulting in a continuous modulation of their membrane potential, due to the occurrence of EPSPs and IPSPs. During fictive locomotor-like activity and after elimination of the cerebellar afferents to the RNm, it was observed that the spontaneous PSPs in RNm cells (in the case of 45% of the cells) were organized in repetitive subthreshold bursts occurring in phase relationships with the activity recorded in the motor nerves. Some extracellularly recorded cells (12%) showed a rhythmic firing pattern. It is generally recognized that, in the thalamic cat preparation, the locomotor pattern observed in efferent nerves originates from the central pattern generator (CPG) of the spinal cord. It therefore seems likely that the rhythmicity observed here in the RNm may originate from the spinal CPG and be transmitted through the spino-rubral pathway ascending in the ventral part of the cord. It is concluded that the spino-rubral pathway may transmit both somatosensory information and corollary discharges relating to the activity of the spinal CPG for locomotion.


Adrenaline in out-of-hospital ventricular fibrillation. Does it make any difference?
A large proportion of cardiac arrests outside hospital are caused by ventricular fibrillation. Although it is frequently used, the exact role of treatment with adrenaline in these patients remains to be determined. To describe the proportion of patients with witnessed out-of-hospital cardiac arrest found in ventricular fibrillation who survived and were discharged from hospital in relation to whether they were treated with adrenaline prior to hospital admission. All the patients with out-of-hospital cardiac arrest found in ventricular fibrillation in Göteborg between 1981 and 1992 in whom cardiopulmonary resuscitation (CPR) was initiated by our emergency medical service (EMS). During the observation period, some of the EMS staff were authorized to give medication and some were not. In all, 1360 patients were found in ventricular fibrillation and detailed information was available in 1203 cases (88%). Adrenaline was given in 417 cases (35%). Among patients with sustained ventricular fibrillation, those who received adrenaline experienced the return of spontaneous circulation more frequently (P < 0.001) and were hospitalized alive more frequently (P < 0.01). However, the rate of discharge from hospital did not differ significantly between the 2 groups. Among patients who converted to asystole or electromechanical dissociation, those who received adrenaline experienced the return of spontaneous circulation more frequently (P < 0.001) and were hospitalised alive more frequently (P < 0.001). However, the rate of discharge from hospital did not differ significantly between the 2 groups. On the basis of 2 treatment regimens during a 12-year survey, we explored the usefulness of adrenaline in out-of-hospital ventricular fibrillation. Both patients with sustained ventricular fibrillation and those who converted to asystole or electromechanical dissociation had an initially more favourable outcome if treated with adrenaline. However, the final outcome was not significantly affected. This study does not confirm the hypothesis that adrenaline increases survival among patients with out-of-hospital cardiac arrest who are found in ventricular fibrillation.


Structural and functional analysis of human liver-expressed antimicrobial peptide 2.
Human liver-expressed antimicrobial peptide 2 (LEAP-2) is a cationic antimicrobial peptide (CAMP) believed to have a protective role against bacterial infection. Little is known about the structure-activity relationships of LEAP-2 or its mechanism of action. In this study we describe the structure of LEAP-2, analyze its interaction with model membranes, and relate them to the antimicrobial activity of the peptide. The structure of LEAP-2, determined by NMR spectroscopy, reveals a compact central core with disorder at the N and C termini. The core comprises a β-hairpin and a 3(10)- helix that are braced by disulfide bonds between Cys17-28 and Cys23-33 and further stabilized by a network of hydrogen bonds. Membrane-affinity studies show that LEAP-2 membrane binding is governed by electrostatic attractions, which are sensitive to ionic strength. Truncation studies show that the C-terminal region of LEAP-2 is irrelevant for membrane binding, whereas the N-terminal (hydrophobic domain) and core regions (cationic domain) are essential. Bacterial-growth-inhibition assays reveal that the antimicrobial activity of LEAP-2 correlates with membrane affinity. Interestingly, the native and reduced forms of LEAP-2 have similar membrane affinity and antimicrobial activities; this suggests that disulfide bonds are not essential for the bactericidal activity. This study reveals that LEAP-2 has a novel fold for a CAMP and suggests that although LEAP-2 exhibits antimicrobial activity under low-salt conditions, there is likely to be another physiological role for the peptide.


Maternal exposure to bisphenol A and anogenital distance throughout infancy: A longitudinal study from Shanghai, China.
Bisphenol A (BPA) is one of the most common endocrine-disrupting compounds (EDCs) with a ubiquitous presence. Both animal and human studies have reported the association between maternal exposure to BPA and anogenital distance (AGD) in offspring. However, the results are conflicting and the longitudinal effect is unknown. We aimed to examine the effect of maternal exposure to BPA on AGD in offspring in a longitudinal birth cohort from birth to 1 year of age. BPA was assayed using urine samples collected at 12-16 gestational weeks from 982 pregnant participants who later delivered infants. Infants' AGDs (AGDap [anus-penis] and AGDas [anus-scrotum] for boys, AGDac [anus-clitoris] and AGDaf [anus-fourchette] for girls) were measured at birth, and at 6 and 12 months of age. Multiple linear regression analysis was conducted to examine the associations between maternal exposure to BPA and offspring's AGDs. Then generalized estimating equation (GEE) model was applied to make use of the repeated measurements of AGDs and examine the overall effect of maternal exposure to BPA. Compared to boys with undetected maternal BPA, those with detected BPA were more likely to have shorter AGDap and AGDas at 6 and 12 months. However, the differences were statistically significant for AGDap and AGDas only at 12 months (2.87 and 4.12 mm shorter, respectively). In GEE models, similar patterns were observed. Boys in the higher quartiles were more likely to have shorter AGDap and AGDas than those in the first quartile. However, statistically significant differences were only observed in boys in the third quartile. For girls, these associations were not observed regardless of the timing of measurements (at birth, 6 months and 12 months). Maternal exposure to BPA was associated with shortened AGDap and AGDas in boys at age 12 months but not in girls, which suggests a gender specific effect of BPA exposure on offspring's development.


The mouse t complex distorter/sterility candidate, Dnahc8, expresses a gamma-type axonemal dynein heavy chain isoform confined to the principal piece of the sperm tail.
Heterozygosity for a t haplotype (t) in male mice results in distorted transmission (TRD) of the t-bearing chromosome 17 homolog to their offspring. However, homozygosity for t causes male sterility, thus limiting the spread of t through the population at large. The Ca(2+)-dependent sperm tail curvature phenotypes, "fishhook", where abnormally high levels of sperm exhibit sharp bends in the midpiece, and "curlicue", where motile sperm exhibit a chronic negative curving of the entire tail, have been tightly linked to t-associated male TRD and sterility traits, respectively. Genetic studies have indicated that homozygosity for the t allele of Dnahc8, an axonemal gamma-type dynein heavy chain (gammaDHC) gene, is partially responsible for expression of "curlicue"; however, its involvement in "fishhook"/TRD, if any, is unknown. Here we report that the major isoform of DNAHC8 is copiously expressed, carries an extended N-terminus and full-length C-terminus, and is stable and equally abundant in both testis and sperm from +/+ and t/t animals. By in silico analysis we also demonstrate that at least three of the seventeen DNAHC8(t) mutations at highly conserved positions in wild-type DHCs may be capable of substantially altering normal DNAHC8 function. Interestingly, DNAHC8 is confined to the principal piece of the sperm tail. The combined results of this study suggest possible mechanisms of DNAHC8(t) dysfunction and involvement in "curlicue", and support the hypothesis that "curlicue" is a multigenic phenomenon. They also demonstrate that the accelerated "fishhook" phenotype of sperm from +/t males is not directly linked to DNAHC8(t) dysfunction.


Synthesis and characterization of alginate/poly-L-ornithine/alginate microcapsules for local immunosuppression.
Alginate/poly-L-ornithine/alginate (APA) coherent microencapsulation, which provides an immunoselective and highly biocompatible membrane, creates a viable option for cellular or tissue transplantation. This study explored the potential of incorporating immunosuppressive drugs onto the capsule surface to provide local immunosuppression in addition to immunoisolation. A thorough investigation has been conducted to optimize and characterize alginate biotinylation via carbodiimide chemistry by a 4'-hydroxyazobenzene-2-carboxylic acid (HABA) based assay and by ATR-FTIR, H-NMR and XPS. To minimize the formation of by-product, a theoretical 40% activation of the carboxylic group on the alginate was employed to manufacture an optimal modification of approximately 10% biotinylated alginate. Confocal fluorescence microscopy was used to assess the conjugation of streptavidin and assembly of antibodies on the microcapsules. Local immunosuppressive capacity was assimilated on the APA microcapsules by binding of anti-tumour necrosis factor-alpha (TNF-alpha) antibodies via streptavidin-biotin conjugation, shown from the clear reduction of TNF-alpha in in-vitro medium.


Effects of blood flow restricted exercise training on muscular strength and blood flow in older adults.
In young adults, blood flow restricted exercise (BFRE) at relatively low intensities can increase muscle strength as effectively as conventional high intensity training. Ischemic exercise can also increase collateral blood flow in skeletal muscle. However, the effects of chronic BFRE on muscle strength and blood flow in older adults remain unknown. The purpose of this study was to compare the effects of 4weeks of BFRE training on skeletal muscle strength and blood flow between young and older subjects and between older adults performing BFRE and conventional high intensity resistance exercise. Maximum voluntary contraction (MVC), forearm girth, peak forearm blood flow (FBF) and forearm vascular conductance (FVC) were assessed before and after 4weeks of forearm resistance training with BFRE in older adults (O-BFRE, 63±1 y, n=9) and younger adults (Y-BFRE, 22±1 y, n=8) and with high intensity training at 75% maximum voluntary contraction in older adults (O-HI, 63±1 y, n=10). MVC increased in all groups (O-BFRE, 33.4±4.7 to 36.3±4.7kg; Y-BFRE, 37.2±4.9 to 43.0±5.0kg; O-HI, 34.0±4.4 to 39.8±4.4kg; all p<0.05). Forearm girth increased in O-BFRE (26.3±1.1 to 26.7±1.1cm; p<0.05) and Y-BFRE (23.9±0.9 to 25.1±1.5cm; p<0.05) but not in O-HI (25.9±1.0 to 26.1±1.0cm; p=0.26). Peak forearm vascular conductance increased in Y-BFRE (0.190±0.016 to 0.311±0.031units; p=0.01) but not in O-BFRE (0.157±0.024 to 0.193±0.029units; p=0.48) and O-HI (0.188±0.035 to 0.227±0.035units; p=0.18). These data suggest that chronic BFRE training is effective in increasing muscular strength, muscle size and vascularity in young adults but, in older adults, increases only muscular strength and size. Longer training durations or higher volumes may be required to evoke similar vascular adaptations in older adults.


The association between hypothyroidism and perinatal outcomes in patients with polycystic ovary syndrome.
To compare pregnancy, delivery, and neonatal outcomes in patients with polycystic ovary syndrome (PCOS) with and without concomitant hypothyroidism. A retrospective population-based cohort study including all women with an ICD-9 diagnosis of PCOS in the US between 2004 and 2014, who delivered in the third trimester or had a maternal death. We compared women with a concomitant diagnosis of hypothyroidism to those without. Women with hyperthyroidism were excluded. Pregnancy, delivery, and neonatal outcomes were compared between the two groups. Overall, 14,882 women met inclusion criteria. Among them, 1882 (12.65%) had a concomitant diagnosis of hypothyroidism, and 13,000 (87.35%) did not. Women with concomitant hypothyroidism, compared to those without, were characterized by increased maternal age (25.5% ≥ 35 years vs. 18%, p < 0.001, respectively), and had a higher rate of multiple gestations (7.1% vs. 5.7%, p = 0.023). Interestingly, pregnancy, delivery and neonatal outcomes were comparable between the groups, except for a higher rate of small-for-gestational-age (SGA) neonates in the group with hypothyroidism (4.1% vs. 3.2%, p = 0.033) (Tables 2 and 3). In a multivariate logistic regression adjusting for potential confounders, hypothyroidism was no longer found to be associated with SGA (adjusted odds ratio [aOR] 1.32, 95% confidence interval [CI] 0.99-1.75, p = 0.057), but was found to increase the odds for preeclampsia (aOR 1.30, 95% CI 1.06-1.59, p = 0.012). In patients with PCOS, concomitant hypothyroidism significantly increases the risk for preeclampsia. Unexpectedly, other pregnancy complications commonly increased by hypothyroidism were not increased in women with PCOS, likely due to the inherent elevated baseline pregnancy risks of PCOS.


Comparison and optimization of strategies for a more profound profiling of the sialylated N-glycoproteomics in human plasma using metal oxide enrichment.
Glycosylation is an important posttranslational modification of proteins and plays a crucial role in both cellular functions and secretory pathways. Sialic acids (SAs), a family of nine-carbon-containing acidic monosaccharides, often terminate the glycan structures of cell surface molecules and secreted glycoproteins and perform an important role in many biological processes. Hence, a more profound profiling of the sialylated glycoproteomics may improve our knowledge of this modification and its effects on protein functions. Here, we systematically investigated different strategies to enrich the SA proteins in human plasma using a newly developed technology that utilizes titanium dioxide for sialylated N-glycoproteomics profiling by mass spectrometry. Our results showed that using a combination of a filter-aided sample preparation method, TiO2 chromatography, multiple enzyme digestion, and two-dimensional reversed-phase peptide fractionation led to a more profound profiling of the SA proteome. In total, 982 glycosylation sites in 413 proteins were identified, among which 37.8% were newly identified, to establish the largest database of sialic acid containing proteins from human plasma.