Association between sleep duration and sarcopenia among community-dwelling older adults: A cross-sectional study.
Both sleep disorders and sarcopenia are common among older adults. However, little is known about the relationship between these 2 conditions.This study aimed to investigate the possible association between sleep duration and sarcopenia in a population of Chinese community-dwelling older adults.Community-dwelling older adults aged 60 years or older were recruited. Self-reported sleep duration, anthropometric data, gait speed, and handgrip strength were collected by face-to-face interviews. Sarcopenia was defined according to the recommended algorithm of the Asian Working Group for Sarcopenia (AWGS).We included 607 participants aged 70.6 ± 6.6 years (range, 60-90 years) in the analyses. The prevalence of sarcopenia in the whole study population was 18.5%. In women, the prevalence of sarcopenia was significantly higher in the short sleep duration group (< 6 hours) and long sleep duration group (>8 hours) compared with women in the normal sleep duration group (6-8 hours; 27.5%, 22.2% and 13.9%, respectively; P = .014). Similar results were found in men; however, the differences between groups were not statistically significant (18.5%, 20.6%, and 13.0%, respectively; P = .356). After adjustments for the potential confounding factors, older women having short sleep duration (OR: 4.34; 95% CI: 1.74-10.85) or having long sleep duration (OR: 2.50; 95% CI: 1.05-6.99) had greater risk of sarcopenia compared with women having normal sleep duration. With comparison to men with normal sleep duration, the adjusted OR for sarcopenia was 2.12 (0.96-8.39) in the short sleep duration group and 2.25 (0.88-6.87) in the long sleep duration group, respectively.A U-shape relationship between self-reported sleep duration and sarcopenia was identified in a population of Chinese community-dwelling older adults, especially in women.


Dissection of the antigenic determinants expressed on the cell surface of RSV-transformed fibroblasts by monoclonal antibodies.
Fusion of P3/X63-Ag8 mouse myeloma cells with splenocytes obtained from mice hyperimmunized with a BHK hamster fibroblast line transformed by an env-strain of Rous sarcoma virus (RSV) resulted in the production of antibody-secreting hybridomas. Seven hybrid clones secreted antibodies binding to RSV-transformed BHK fibroblasts but not to the parental control non-transformed line. The antibodies produced by three of these clones did identify antigenic determinants expressed also on BHK cells transformed by SV 40. The antibodies produced by four other clones reacted specifically with cells transformed by RSV but not with cells productively infected by the transformation-defective Rous-associated virus-1; one of these reacted with RSV-transformed hamster cells only, three others identified antigenic determinants common to RSV-transformed cells of different animal species. These data give further support to the idea that RSV-transformed cells express a cell-surface antigen, specific for transformation, the expression of which is controlled by the transforming src gene, and that the specificities carried on it follow a complex pattern, arising from the interaction between antigenic determinants of cellular and viral origin.


Inhibitors of the Hepatitis C Virus Polymerase; Mode of Action and Resistance.
The hepatitis C virus (HCV) is a pandemic human pathogen posing a substantial health and economic burden in both developing and developed countries. Controlling the spread of HCV through behavioural prevention strategies has met with limited success and vaccine development remains slow. The development of antiviral therapeutic agents has also been challenging, primarily due to the lack of efficient cell culture and animal models for all HCV genotypes, as well as the large genetic diversity between HCV strains. On the other hand, the use of interferon-α-based treatments in combination with the guanosine analogue, ribavirin, achieved limited success, and widespread use of these therapies has been hampered by prevalent side effects. For more than a decade, the HCV RNA-dependent RNA polymerase (RdRp) has been targeted for antiviral development, and direct-acting antivirals (DAA) have been identified which bind to one of at least six RdRp inhibitor-binding sites, and are now becoming a mainstay of highly effective and well tolerated antiviral treatment for HCV infection. Here we review the different classes of RdRp inhibitors and their mode of action against HCV. Furthermore, the mechanism of antiviral resistance to each class is described, including naturally occurring resistance-associated variants (RAVs) in different viral strains and genotypes. Finally, we review the impact of these RAVs on treatment outcomes with the newly developed regimens.


The association of self-injurious behaviour and suicide attempts with recurrent idiopathic pain in adolescents: evidence from a population-based study.
While several population-based studies report that pain is independently associated with higher rates of self-destructive behaviour (suicidal ideation, suicide attempts, and self-injurious behaviour) in adults, studies in adolescents are rare and limited to specific chronic pain conditions. The aim of this study was to investigate the link between self-reported idiopathic pain and the prevalence and frequency of self-injury (SI) and suicide attempts in adolescents. Data from a cross-sectional, school-based sample was derived to assess SI, suicide attempts, recurrent pain symptoms and various areas of emotional and behavioural problems via a self-report booklet including the Youth Self-Report (YSR). Adolescents were assigned to two groups (presence of pain vs. no pain) for analysis. Data from 5,504 students of 116 schools in a region of South Western Germany was available. A series of unadjusted and adjusted multinomial logistic regression models were performed to address the association of pain, SI, and suicide attempts. 929 (16.88%) respondents reported recurrent pain in one of three areas of pain symptoms assessed (general pain, headache, and abdominal pain). Adolescents who reported pain also reported greater psychopathological distress on all sub-scales of the YSR. The presence of pain was significantly associated with an increased risk ratio (RR) for SI (1-3 incidences in the past year: RR: 2.96; >3 incidences: RR: 6.04) and suicide attempts (one attempt: RR: 3.63; multiple attempts: RR: 5.4) in unadjusted analysis. Similarly, increased RR was observed when adjusting for sociodemographic variables. While controlling for psychopathology attenuated this association, it remained significant (RRs: 1.4-1.8). Sub-sequent sensitivity analysis revealed different RR by location and frequency of pain symptoms. Adolescents with recurrent idiopathic pain are more likely to report previous incidents of SI and suicide attempts. This association is likely mediated by the presence of psychopathological distress as consequence of recurrent idiopathic pain. However, the observed variance in dependent variables is only partially explained by emotional and behavioural problems. Clinicians should be aware of these associations and interview adolescents with recurrent symptoms of pain for the presence of self-harm, past suicide attempts and current suicidal thoughts. Future studies addressing the neurobiology underpinnings of an increased likelihood for self-injurious behaviour and suicide attempts in adolescents with recurrent idiopathic pain are necessary.


Development of a video-based evaluation tool in Rett syndrome.
This paper describes the development of a video-based evaluation tool for use in Rett syndrome (RTT). Components include a parent-report checklist, and video filming and coding protocols that contain items on eating, drinking, communication, hand function and movements, personal care and mobility. Ninety-seven of the 169 families who initially agreed to participate returned a videotape within 8 months of the first request. Subjects whose videos were returned had a similar age profile to those who did not provide a video but were more likely to have classical than atypical RTT. Evidence of the content and social validity and inter-rater reliability on 11 videos is provided. Video may provide detailed, objective assessment of function and behaviour in RTT.


Antidepressant & anxiolytic activities of N-(pyridin-3-yl) quinoxalin-2-carboxamide: A novel serotonin type 3 receptor antagonist in behavioural animal models.
Alteration in the serotonin leads to the psychological illness, such as depression, anxiety, schizophrenia, eating disorders, obsessive-compulsive disorder, panic disorders and migraines. The objective of the current study was to investigate the antidepressant and anxiolytic activities of N-(pyridin-3-yl) quinoxalin-2-carboxamide (QCF-21), a novel 5-HT3receptor antagonist in preclinical models of depression and anxiety. Antidepressant activity was evaluated in preliminary tests such as forced swim and tail suspension tests (FST & TST). Anti-anxiety effect of QCF-21 was investigated by employing elevated plus maze (EPM), light/dark and hole board tests. Olfactory bulbectomy (OBX) in rats was used as chronic model of depression. Mechanistic test of QCF-21 was evaluated by reserpine-induced hypothermia and 5-hydroxytryptophan (5-HTP)-induced head-twitch response. The dose-response study revealed an initial antidepressant-like effect of QCF-21(0.25-1 mg/kg, i.p.) in the FST and TST and anxiolytic-like effect in EPM, light and dark and hole board tests. QCF-21 potentiated the 5-HTP-induced head-twitches response in mice and reversed reserpine-induced hypothermia in rats. QCF-21 significantly reversed the behavioural anomalies post-OBX in rats. The present findings indicate the potential antidepressant-like and anxiolytic-like effects of QCF-21 at low doses in rodent behavioural models of depression and anxiety. Further studies need to be done to understand the underlying mechanism.


Effects of porcine reproductive and respiratory syndrome virus-infected antigen-presenting cells on T cell activation and antiviral cytokine production.
The ability of porcine reproductive and respiratory syndrome virus (PRRSV) to suppress T cell expression of CD25 (alpha chain of interleukin [IL]-2 receptor), interferon-gamma (IFN-gamma), and tumor necrosis factor-alpha (TNF-alpha) was determined by flow cytometry in naive porcine T cells in response to mitogen (concanavalin A) and cytokine inducers (phorbol 12-myristate 13-acetate plus ionomycin [PMA/I]). Four PRRSV isolates of varying clinical virulence and three different types of porcine myeloid antigen-presenting cells (APCs) were used. T cells cultured with monocytes infected with virulent PRRSV (VR-2385, SDSU-73, and VR-2332), but not with a vaccine strain (Ingelvac PRRS MLV; Boehringer Ingelheim Vetmedica, St. Joseph, MO), demonstrated significantly reduced CD25 expression (%CD25(+)) and IFN-gamma expression (%IFN-gamma (+)) compared with T cells incubated with uninoculated monocyte cultures. T cells cultured with monocytes infected with all four PRRSV isolates demonstrated significantly reduced %TNF-alpha (+). The significant reduction of %CD25(+), %IFN-gamma (+), and %TNF-alpha (+) was not detected in T cells cultured with monocyte-derived macrophages (MDMs) and immature monocyte-derived dendritic cells (MDCs) infected with any PRRSV isolates. Heat-inactivated PRRSV did not induce significantly reduced T cell responses in any APC cultures. The reduction of T cell response in monocyte cultures was not due to PRRSV-induced T cell death. Gene expression of IL-10 detected by semiquantitative reverse transcriptase-polymerase chain reaction was significantly increased in virulent PRRSV-infected monocyte cultures after PMA/I, but not concanavalin A, stimulation compared with IL-10 gene expression from uninoculated monocyte cultures. Increased IL-10 gene expression contributed to significantly reduced %IFN-gamma (+) and %TNF-alpha (+), but not %CD25(+), as determined by IL-10 neutralization assay. This study reports that PRRSV has the ability to suppress T cell responses. The suppressive ability of PRRSV is associated with viral virulence and is mediated by virus-infected monocytes, but not by virus-infected MDMs and immature MDCs.


Dipyridamole reverses the resistance to topoisomerase II inhibitors but not to antimicrotubule agents in multidrug-resistant melanoma cells.
The influence of dipyridamole (DP) on the cytotoxicity and cellular disposition of several DNA topoisomerase II (topo II) inhibitors and antimitotic agents in multidrug-resistant B16VDXR cells was examined. B16VDXR cells, derived from parental B16V cells by step-wise treatment with doxorubicin (DOX), overexpress a 170 kDa P-glycoprotein (P-gp). Additionally, the resistance to DOX in B16VDXR cells is associated with decreased frequency of DNA strand breaks compared to that in the drug-sensitive B16V cells. DP (10 microM) significantly (P < 0.01) potentiated the cytotoxicity of DOX (6.4-fold), mitoxantrone (2.3-fold), and etoposide (14-fold) in the drug-resistant B16VDXR cells. This was accompanied by a 3.7-fold and 4.2-fold increase in the total intracellular and nuclear levels of DOX, respectively. Surprisingly, no significant change in the intracellular and nuclear levels or the efflux of etoposide was observed in B16VDXR cells. Combination index (CI) analysis, however, indicated that DP interacted synergistically with DOX as well as etoposide. Further, it was intriguing to observe that DP (10 microM) failed to modulate the resistance to vincristine, vinblastine, and taxol. This was despite a significant increase in the accumulation of vinblastine (3.3-fold) and taxol (3.9-fold) in B16VDXR cells in the presence of DP (10 microM). The observed pattern of chemosensitization suggests that in addition to interaction with P-gp, the multidrug-resistance modulating activity of DP may involve P-gp independent mechanism(s). The possibilities include that (i) DP interacts with topo II or (ii) DP promotes the formation and/or obstructs the repair of DNA strand breaks caused by topo II inhibitors.


Incorporation of Molecular Catalysts in Metal-Organic Frameworks for Highly Efficient Heterogeneous Catalysis.
Porous metal-organic frameworks (MOFs) are built from periodically alternate organic moieties and metal ions/clusters. The unique features of the open framework structures, the high surface areas, the permanent porosity, and the appropriate hydrophilic and hydrophobic pore nature mean that MOF materials are a class of ideal host matrices for immobilization of molecular catalysts. The emerging porous materials can not only retain but are also able to enhance the catalytic functions of the single individuals. MOF catalysts have the following super characters: i) uniformly dispersed catalytic sites on the pore surfaces to improve the utility, ii) appropriate hydrophilic and hydrophobic pore nature to facilitate the recognition and transportation of reactant and product molecules, iii) a collaborative microenvironment to realize synergistic catalysis, and iv) simple separation and recovery for long-term usage. Accompanying the development of the synthetic strategies and the technologies for the characterization of MOF materials, MOF catalysis has undergone an upsurge, which has transcended the stage of opportunism. Here, the rational design and synthesis of MOF catalysts are discussed, along with the key factors of active sites, microenvironments, and transmission channels that lead to the distinct catalytic properties of MOF catalysts.


Association of Impaired Reactive Aldehyde Metabolism with Delayed Graft Function in Human Kidney Transplantation.
Delayed graft function is an early complication following kidney transplantation with an unclear molecular mechanism. Here we determined whether impaired reactive aldehyde metabolism is associated with delayed graft function. Human kidney biopsies from grafts with delayed graft function were compared with grafts that did not develop delayed graft function by Ingenuity gene pathway analysis. A second series of grafts with delayed graft function (n = 10) were compared to grafts that did not develop delayed graft function (n = 10) by measuring reactive aldehyde metabolism, reactive aldehyde-induced protein adduct formation, and aldehyde dehydrogenase (ALDH) gene and protein expression. In the first series of kidney biopsies, several gene families known for metabolizing reactive aldehydes, such as aldehyde dehydrogenase (ALDH), aldo-keto reductase (AKR), and glutathione-S transferase (GSTA), were upregulated in kidneys that did not develop delayed graft function versus those that did. In the second series of kidney grafts, we focused on measuring aldehyde-induced protein adducts and ALDH enzymatic activity. The reactive aldehyde metabolism by ALDH enzymes was reduced in kidneys with delayed graft function compared to those that did not (37 ± 12∗ vs. 79 ± 5 μg/min/mg tissue, ∗ P < 0.005, respectively). ALDH enzymatic activity was also negatively correlated with length of hospital stay after a kidney transplant. Together, our study identifies a reduced ALDH enzymatic activity with kidneys developing delayed graft function compared to those that did not. Measuring ALDH enzymatic activity and reactive aldehyde-induced protein adducts can potentially be further developed as a biomarker to assess for delayed graft function and recovery from a kidney transplant.


A comparison of free-hand vs laser-guided long-axis ultrasound techniques in novice users.
The increasing use of point-of-care ultrasonography for targeted procedures justifies a device that helps both novices in training and experts perform the long-axis needle approach. The initial success of traditional needle guidance devices in reducing the time of target procedures is not universal and they can be cumbersome. We aim to investigate whether the less bulky and previously untested laser guide can succeed in reducing procedure time in novice ultrasonographers. 82 medical students with no ultrasound experience volunteered. Random allocation determined whether, during a targeted procedure in a turkey breast and olive phantom, participants were assisted by the laser guide or not. The time taken to pierce the target was recorded at 1-cm depth. The mean procedure time in the laser-assisted (LA) group was 25.1 s (14.0 s; 18.0-25.0 s). The mean procedure time in the free-hand group was 45.5 s (23.0 s; 7.0-55.0 s). The procedure time in the LA group was significantly reduced (p<0.01). The laser guide significantly improved procedure times. It is felt that the cheaper, smaller, easy to integrate, sterile and more user-friendly laser guidance unit may be a better alternative to the needle guide in improving procedure times for the novice ultrasonographer or to assist the expert, during training for, or performance of, ultrasound-guided targeted procedures. Following from the prototype paper, this is the first study to investigate the effectiveness of attaching a laser-guidance device to an ultrasound probe. The device succeeded in reducing the procedure times of targeted procedures.


Nucleolar- and Nuclear-Stress-Induced Membrane-Less Organelles: A Proteome Analysis through the Prism of Liquid-Liquid Phase Separation.
Radical changes in the idea of the organization of intracellular space that occurred in the early 2010s made it possible to consider the formation and functioning of so-called membrane-less organelles (MLOs) based on a single physical principle: the liquid-liquid phase separation (LLPS) of biopolymers. Weak non-specific inter- and intramolecular interactions of disordered polymers, primarily intrinsically disordered proteins, and RNA, play a central role in the initiation and regulation of these processes. On the other hand, in some cases, the "maturation" of MLOs can be accompanied by a "liquid-gel" phase transition, where other types of interactions can play a significant role in the reorganization of their structure. In this work, we conducted a bioinformatics analysis of the propensity of the proteomes of two membrane-less organelles, formed in response to stress in the same compartment, for spontaneous phase separation and examined their intrinsic disorder predispositions. These MLOs, amyloid bodies (A-bodies) formed in the response to acidosis and heat shock and nuclear stress bodies (nSBs), are characterized by a partially overlapping composition, but show different functional activities and morphologies. We show that the proteomes of these biocondensates are differently enriched in proteins, and many have high potential for spontaneous LLPS that correlates with the different morphology and function of these organelles. The results of these analyses allowed us to evaluate the role of weak interactions in the formation and functioning of these important organelles.


Increased contractility of isolated lung parenchyma in an animal model of pulmonary fibrosis induced by bleomycin.
The contractile capability of lung parenchymal strips isolated from normal rats was compared with that of strips isolated from rats with pulmonary fibrosis induced by intratracheal instillation of bleomycin sulfate. Subsequently, the population of contractile cells, both muscle and nonmuscle, was analyzed in each strip by histochemical, immunocytochemical, and ultrastructural means. The force (g force/g tissue wet weight) generated by fibrotic strips was approximately double that of the control strips in response to acetylcholine, epinephrine, and potassium depolarizing solution. The content of smooth muscle in fibrotic and control strips was not significantly different. Immunofluorescent examination indicated increased contractile proteins (actin and myosin) in the thickened alveolar walls of the fibrotic strips in areas devoid of histologically demonstrable smooth muscle. Ultrastructural examination of the fibrotic interstitium revealed an increased population of filament-laden cells that appeared related to the contractile interstitial cell, or myofibroblast. The results indicate that parenchymal strips isolated from fibrotic rat lung can generate increased force in vitro. These responses may be related to increased nonmuscle contractile cells in the fibrotic interstitium.


Preparation and characterization of antioxidant edible chitosan films incorporated with epigallocatechin gallate nanocapsules.
Antioxidant edible films based on chitosan hydrochloride (CHC) and epigallocatechin gallate (EGCG)-loaded nanocapsules (NCs) were developed. The CHC films incorporated with NCs were prepared by a casting method in three different proportions. The obtained films were characterized using the techniques including scanning electron microscopy, Fourier transform infrared spectroscopy and X-ray diffraction. Meanwhile, the mechanical and color properties, optical transmittance, EGCG release profile and antioxidant activity were also determined. The addition of NCs to CHC films increased their tensile strength (TS) and the percent elongation at break (%E). Lightness was significantly decreased in the NCs embedded films compared to controls, which in turn led to high antioxidant activity and excellent barrier properties against visible light. EGCG was rapidly released into food simulant although the concentration fluctuates over time. Therefore, these films can prevent the oxidation of fatty food stuffs.


[Intramural aortic hematoma].
Aortic dissection is a medical and/or surgical emergency that is usually catastrophic if not diagnosed and treated promptly. Transesophageal Doppler echography provides an accurate method of diagnosing and evaluating dissection of aortic aneurysm. Due to the high resolution of transesophageal echocardiography, special subtypes of aortic dissection such as intramural hemorrhage can be diagnosed. The purpose of this paper is to report a case and review the anatomical, clinical and transesophageal echocardiography aspects of aortic dissection without intimal rupture (intramural hematoma).


Beta factor in a random laser.
We develop a definition for the beta factor, the fraction of spontaneous emission that seeds the laser process, for a random laser. With the wavelength-dependence of the gain (and potentially scattering) being the only possible criterion in the competition between gain and loss, our concept of beta is based on the spectral properties of the spontaneous emission and laser light. We find beta approximately equal to 0.1. We discuss the apparent similarities and differences between the beta for a cavity and a random laser.


High-fat diets promote colon orthotopic transplantation tumor metastasis in BALB/c mice.
High-fat diets (HFDs) are a risk factor for colorectal cancer. The present study investigated whether HFDs increase colon cancer metastasis in BALB/c mice. A total of 40 BALB/c mice were divided into four groups, including the tumor, tumor-HFD, HFD and control groups. After 3 weeks, the tumor weights and metastases were observed. The serum levels of triglyceride, total cholesterol, lapin, interleukin-6 (IL-6) and tumor necrosis factor were analyzed using ELISA. The CD34, vascular endothelial growth factor (VEGF) and angiotensin 2 (ANG2) protein and mRNA levels in tumor tissues were analyzed with immunohistochemistry and reverse transcription-polymerase chain reaction. The metastasis frequency increased in the tumor-HFD group. However, there was no difference in the mean tumor weight between the tumor-HFD and tumor groups. The serum cholesterol levels were increased in the tumor-HFD and HFD groups compared with the control group. The levels of serum IL-6 and tumor necrosis factor-α were increased in the tumor-HFD group compared with other groups. The CD34 protein level, and VEGF protein and mRNA levels were increased in the tumor-HFD group compared with the tumor group. No difference was identified between the ANG2 protein and mRNA levels in of the two groups. It was concluded that HFD increased the serum level of cholesterol and cytokines, and potentially induced tumor angiogenesis, promoting transplanted orthotopic colon tumor metastasis in BALB/c mice.


Transvenous magnetic tip lead system for permanent pacing of the heart. A preliminary report.
An electrode for transvenous permanent ventricular pacing equipped with a supermagnet was implanted in six dogs in the apex of the right ventricle for several weeks. As no signs of myocardial perforation occurred, the device which consists of the magnetic tip lead and an external supermagnet applied to the precordial region of the chest wall was implanted in 14 patients. The lead was introduced through the right cephalic vein in 12 patients and through the right external jugular vein in the remaining 2 patients. Threshold values were measured twice a day in 12 patients who received a Siemens-Elema Vario pacemaker 629. The average peak threshold was reached on the 11th day after implantation. No pacing failure or other signs of acute dislodgement of leads occurred during the first 30 days.


Granulocyte colony-stimulating factor-producing undifferentiated carcinoma of the colon mimicking a pulmonary giant cell carcinoma: a case showing overexpression of CD44 along with highly proliferating nestin-positive tumor vessels.
Granulocyte colony-stimulating factor (G-CSF)-producing tumors are known for their aggressive behavior. Only four cases of G-CSF-producing colorectal carcinoma have been previously reported. Herein, we present a case of an undifferentiated carcinoma of the descending colon showing G-CSF production and giant cell carcinoma morphology in a 93-year-old woman. A tumor with a diameter of 80 mm was identified in the descending colon via computed tomography. Descending colectomy was performed involving the abdominal wall where tumor invasion was observed. The white blood cell count, which was elevated before resection, decreased to normal levels after intervention. However, local recurrence at the resected site was detected 39 days after surgery. Upon recurrence, increased white blood cell counts and serum G-CSF were seen. The patient died because of respiratory failure 98 days after colectomy. By using immunohistochemistry, G-CSF expression was detected in tumor cells in the resected specimen, along with overexpression of CD44 and highly proliferating nestin-positive tumor vessels. The poor clinical outcome of this patient is consistent with previous reports that the expression of these three molecules predict poor prognosis. While G-CSF can be a therapeutic target considering its auto/paracrine function to induce tumor growth via the G-CSF receptor, CD44 and nestin may also be possible candidate therapeutic targets. Further studies are required to assess the efficacy of treatments targeting these three molecules.


Congress on women's health Trudy Bush lecture 2014: new insights into sex Hormones and Cardiovascular disease.
Many basic, mechanistic studies of how sex steroids alter vascular function proceeded from Dr. Bush's seminal epidemiologic observations that noncontraceptive use of estrogen reduced all-cause and cardiovascular mortality. However, translating findings from these basic science studies into clinical trials and clinical guidelines has been controversial. This commentary reviews the development of sex steroid vascular research, identifies mechanisms by which sex steroids affect vascular function, reviews findings from recent clinical hormone trials, and identifies challenges and the need for continued funding of such investigations.


Temporal references and temporal relations in sentence comprehension.
The author investigated the interpretation of temporal references during comprehension of sentences containing a main and subordinate clause. Experiments 1 and 2 examined state and event subordinate clauses, respectively, and showed that subordinate temporal references overlapping with or close to the time of the main clause event were read faster than nonoverlapping distant references. Experiment 3 examined temporal references in nonsubordinate main clauses and confirmed that temporal relations between main and subordinate clauses were established on-line in the previous experiments. Experiment 4 independently manipulated temporal overlap and distance and suggested that event and state clauses are processed according to distinct temporal parameters. The results are explained by the contingency relations that events and states establish with other discourse events.


Pulmonary blastoma with fatal hypercalcemia.
We describe a 50-year-old, previously healthy male with metastatic pulmonary blastoma associated with hypercalcemic and hyperosmolar complications which caused his death after 5 days. The primary tumor consisted of epithelial [cytokeratin, beta-hCG CEA, neuron-specific enolase (NSE)-positive] and mesenchymal components (beta-hCG, vimentin, NSE-positive, while the metastases had only a mesenchymal component.


Development of hepatitis B virus capsids into a whole-chain protein antigen display platform: new particulate Lyme disease vaccines.
The immunogenicity of peptides and small protein fragments can be considerably enhanced by their presentation on particulate carriers such as capsid-like particles (CLPs) from hepatitis B virus (HBV). HBV CLPs are icosahedral nanoparticles formed by 90 or 120 core protein dimers. Insertions into the immunodominant c/e1 B cell epitope, a surface-exposed loop on the HBV capsid protein, are especially immunogenic. Here we investigated whether the HBV core protein can be exploited as a vaccine carrier for whole-chain protein antigens, using two clinically relevant proteins derived from a bacterial human pathogen, the Lyme disease agent Borrelia burgdorferi. For this purpose we analyzed CLP formation by core fusions with the entire 255-amino-acid ectodomain of outer surface lipoprotein A (OspA), and with two distinct, 189 amino acid long variants of the dimeric OspC (OspC(a), OspC(b)) of B. burgdorferi. OspA appropriately inserted into the HBV core protein yielded a multimerization-competent fusion protein, termed coreOspA. Although only partially assembling into regular CLPs, coreOspA induced antibodies to OspA, including the Ig isotype profile and specificity for the protective epitope "LA-2", with an efficiency similar to that of recombinant lipidated OspA, the first generation vaccine against Lyme disease. Moreover, coreOspA actively and passively protected mice against subsequent challenge with B. burgdorferi. Fusions with the two OspC variants were found to efficiently form regular CLPs, most probably by OspC dimerization across different core protein dimers. In mice, both coreOspC preparations induced high-titered antibody responses to the homologous but also to the heterologous OspC variant, which conferred protection against challenge with B. burgdorferi. The data demonstrate the principal applicability of HBV CLPs to act as potent immunomodulator even for structurally complex full-length polypeptide chains, and thus open new avenues for novel vaccine designs.


Polygraph testing for deception in Australia: effective aid to crime investigation and adjudication?
Polygraph testing--or the monitoring and analysing of selected physiological measures of an individual who is being interviewed, for the purpose of detecting deception--is controversial in Australia. Considerable mythology surrounds this method of detecting deception. Embedded in popular perceptions of crime fighting and utilised in high-profile criminal cases in Western Australia, Victoria and Queensland, polygraph testing is also explicitly prohibited from being used in crime investigation in New South Wales. Unlike in the United States, polygraph testing has not hitherto routinely been used by government departments and authorities as a preemployment screening tool, but is increasingly being offered in the private sector in Australia. This article examines the current scope of polygraph testing in Australia, describes different approaches to testing, briefly reviews recent information relating to validity and evaluates State legislation prohibiting the use of polygraph testing for specified purposes in New South Wales. Consequent to the continuing controversy regarding the accuracy of polygraph testing in detecting deception (and, conversely, truth-telling), it is argued that the emerging use of polygraph testing is problematic and common law principles rendering the results of such testing inadmissible in court do not constitute sufficient safeguard against inappropriate and intrusive testing. Future research should identify specific polygraph testing techniques and consider the most socially beneficial way of regulating this emerging area of practice.


Association of Serum Myostatin with Body Weight, Visceral Fat Volume, and High Sensitivity C-Reactive Protein But Not With Muscle Mass and Physical Fitness in Premenopausal Women.
The myokine myostatin regulates muscle mass and has been linked to insulin resistance and metabolic syndrome. However, data on its role in humans is still limited. We, therefore, investigated the associations of serum myostatin with muscle mass, physical fitness, and components of the metabolic syndrome in a cohort of premenopausal women. We undertook a cross-sectional analysis of 233 women from the monocenter study PPSDiab, conducted in Munich, Germany. Participants had recently completed a pregnancy with or without gestational diabetes. Our analysis included medical history, anthropometrics, oral glucose tolerance testing, laboratory chemistry, cardiopulmonary exercise testing, and magnetic resonance imaging (n=142) of visceral fat volume, left quadriceps muscle mass, and muscle fat content. Serum myostatin was quantified by a competitive enzyme-linked immunosorbent assay. We observed positive correlations of serum myostatin with body mass index (ρ=0.235; p=0.0003), body fat percentage (ρ=0.166; p=0.011), waist circumference (ρ=0.206; p=0.002), intraabdominal fat volume (ρ=0.182; p=0.030) and high-sensitivity C-reactive protein (ρ=0.175; p=0.008). These correlations were reproduced in linear regression analyses with adjustment for age and time after delivery. We saw no correlations with muscle mass, physical fitness, insulin sensitivity, triglycerides, HDL cholesterol, and blood pressure. Our observation of elevated serum myostatin in women with a higher body fat percentage, visceral obesity, and elevated c-reactive protein suggests that this myokine contributes to the altered muscle-adipose tissue crosstalk in metabolic syndrome. Elevated myostatin may advance this pathophysiologic process and could also impair the efficacy of exercise interventions. Further mechanistic studies, therefore, seem warranted.


[Transfection of pEGFP-C2 in brain mediated by targeting liposome P-MMA-DOSPER].
This research tried improving the specificity and efficiency of gene transfection in gene therapy and tried making the liposome a better gene transfer vector to brain by use of the monoclonal antibody (anti-Lex/SSEA-1)-mediated targeting of liposome. The derivatized monoclonal antibody was conjugated to the liposome DOSPER to form the targeting liposome P-MMA-DOSPER. Then, the pEGFP-C2 encapsulated in P-MMA-DOSPER or DOSPER was injected into the lateral ventricle of SD rats respectively, and the brains were taken for frosted slice 1, 3, 7 or 14 days later. The expression of GFP was observed under fluorescent microscope. There was a lot of expression of GFP around the lateral ventricle of rats in each group. But the indirect fluorescence antibody test showed the ratio of GFP+/nestin+ cells to nestin+ cells of every marking time point in the group of P-MMA-DOSPER was higher than the one in the group of DOSPER; the difference was found to be statistically significant (P<0.01). The results proved that the P-MMA-DOSPER can permeat the ependyma and can transfer gene into the nerve stem cells in vivo safely and effectively.


Isolation and characterization of gastric microsomal glycoproteins. Evidence for a glycosylated beta-subunit of the H+/K(+)-ATPase.
Detergent-solubilization of hog gastric microsomal membrane proteins followed by affinity chromatography using wheat germ agglutinin or Ricinus communis I agglutinin resulted in the isolation of five glycoproteins with the apparent molecular masses on sodium dodecyl sulfate polyacrylamide gels of (in kDa): 60-80 (two glycoproteins sharing this molecular mass); 125-150; and 190-210. In the nonionic detergent Nonidet P-40 (NP-40), the 94 kDa H+/K(+)-ATPase was recovered exclusively in the lectin-binding fraction; however, in the cationic detergent dodecyltrimethylammonium bromide, most of the ATPase was recovered in the nonbinding fraction. Detection of glycoproteins either by periodic acid-dansyl hydrazine staining of carbohydrate in polyacrylamide gels or by Western blots probed with lectins indicated that the majority of the ATPase molecules are not glycosylated. In addition, in the absence of microsomal glycoproteins, the NP-40-solubilized ATPase does not bind to a lectin column. Taken together, these results suggest that the recovery of NP-40-solubilized ATPase in the lectin-binding fraction is due to its noncovalent interaction with a gastric microsomal glycoprotein. Immunoprecipitation of the ATPase from NP-40-solubilized microsomal membrane proteins resulted in the co-precipitation of a single 60-80 kDa glycoprotein. Characterization of the 60-80 kDa glycoprotein associated with the ATPase revealed that: it is a transmembrane protein; it has an apparent core molecular mass of 32 kDa; and, it has five asparagine-linked oligosaccharide chains. Given its similarity to the glycosylated beta-subunit of the Na+/K(+)-ATPase, this 60-80 kDa gastric microsomal glycoprotein is suggested to be a beta-subunit of the H+/K(+)-ATPase.


Interacting Effects of TSH and Insulin on Human Differentiated Adipocytes.
Subclinical hypothyroidism, characterized by an isolated rise in TSH serum levels with normal thyroid function, is a pro-inflammatory state associated with insulin resistance. Adipocytes express TSH receptors, but it is not known if TSH can directly inhibit insulin signaling. Using primary human differentiated adipocytes, we examined the effects of TSH on insulin-stimulated Akt phosphorylation, and whether conventional PKC (cPKC) were involved. The effect of insulin on TSH-stimulated lipolysis was also investigated. TSH inhibited insulin-stimulated Akt phosphorylation in adipocytes by 54%. TSH activated cPKC, and Gö6976, a PKCα and -β1 inhibitor, prevented the inhibitory effect of TSH on the insulin response. Insulin reduced the ability of TSH to activate cPKC and to stimulate lipolysis.Our data reveal novel interactions between TSH and insulin. TSH inhibits insulin-stimulated Akt signaling in a cPKC-dependent fashion, whereas insulin blocks TSH-stimulated cPKC activity and lipolysis. TSH and insulin act on differentiated human adipocytes to modulate their respective intracellular signals.


Feeding signals inhibit fluid-satiation signals in the mouse lateral parabrachial nucleus to increase intake of highly palatable, caloric solutions.
Chemogenetic activation of oxytocin receptor-expressing neurons in the parabrachial nucleus (OxtrPBN neurons) acts as a satiation signal for water. In this research, we investigated the effect of activating OxtrPBN neurons on satiation for different types of fluids. Chemogenetic activation of OxtrPBN neurons in male and female transgenic OxtrCre mice robustly suppressed the rapid, initial (15-min) intake of several solutions after dehydration: water, sucrose, ethanol and saccharin, but only slightly decreased intake of Ensure®, a highly caloric solution (1 kcal/mL; containing 3.72 g protein, 3.27 g fat, 13.42 g carbohydrates, and 1.01 g dietary fibre per 100 mL). OxtrPBN neuron activation also suppressed cumulative, longer-term (2-h) intake of lower caloric, less palatable solutions, but not highly caloric, palatable solutions. These results suggest that OxtrPBN neurons predominantly control initial fluid-satiation responses after rehydration, but not longer-term intake of highly caloric, palatable solutions. The suppression of fluid intake was not because of anxiogenesis, but because OxtrPBN neuron activation decreased anxiety-like behaviour. To investigate the role of different PBN subdivisions on the intake of different solutions, we examined FOS as a proxy marker of PBN neuron activation. Different PBN subdivisions were activated by different solutions: the dorsolateral PBN similarly by all fluids; the external lateral PBN by caloric but not non-caloric solutions; and the central lateral PBN primarily by highly palatable solutions, suggesting PBN subdivisions regulate different aspects of fluid intake. To explore the possible mechanisms underlying the minimal suppression of Ensure® after OxtrPBN neuron activation, we demonstrated in in vitro slice recordings that the feeding-associated agouti-related peptide (AgRP) inhibited OxtrPBN neuron firing in a concentration-related manner, suggesting possible inhibition by feeding-related neurocircuitry of fluid satiation neurocircuitry. Overall, this research suggests that although palatable beverages like sucrose- and ethanol-containing beverages activate fluid satiation signals encoded by OxtrPBN neurons, these neurons can be inhibited by hunger-related signals (agouti-related peptide, AgRP), which may explain why these fluids are often consumed in excess of what is required for fluid satiation.


The association between socioeconomic status and visual disability among older adults in China.
To investigate the association between socioeconomic status (SES) and visual disability (VD) among older Chinese adults. We obtained data from the Second National Sample Survey on Disability, conducted in China in 2006. A total number of 192 375 older adults (aged≥65y) were screened for suspected VD via interviews with trained examiners. Those who screened positively for VD were referred to ophthalmologists to obtain a final diagnosis. VD was prevalent among 7.29% of Chinese adults aged 65 and older, and was higher in rural areas (8.71%) than in urban areas (4.82%). After adjusting for SES indicators and covariates, we found that less-educated older adults were more likely to suffer from VD, with an odds ratio (OR) of 2.50 (95%CI: 2.26-2.82) for illiterates, compared with those who graduated from senior high school or above. Older adults who were in the lowest income quintile were more at risk of VD, with an OR of 1.81 (95%CI: 1.68-2.95), compared with adults in the highest income quintile. In urban areas, when compared with adults who graduated from senior high school or above, those who did not continue their education after junior high school, primary school, or those who were illiterate, were more likely to suffer from VD, with an OR of 1.35 (95%CI: 1.51-1.59), 1.84 (95%CI: 1.60-2.12), and 2.63 (95%CI: 2.27-3.04), respectively. Lower levels of income were statistically significant when associated with VD. In rural areas, adults who were illiterate had an OR of 2.21 (95%CI: 1.75-2.79) when compared to adults with senior high school or above education level. Per capita, household income remained significantly associated with VD. Older adults who were ≥85, female, single, and residing in rural areas were associated with higher risks of VD. Individual-level SES among the elderly, in the form of education and income, is associated with VD among elderly Chinese adults in both urban and rural areas; however, the association is stronger in rural areas. Further studies are still required to explore the mechanism behind the relationships.


[People, the environment and health: the "Oneness" of human health from the perspective of universal life presented in "Changes"].
This paper aimed to expand the paradigm of nursing and expand the essential factors of nursing theories beyond "environment" to encompass universal life. While individuals live between the sky and earth, we are an inseparable part of the universe. "Health" is derived from a oneness that embraces the body, mind and spirit. The human body contains the wisdom of the universe, known in Chinese philosophy as the wisdom of "Changes". The body has its own consciousness and possesses great powers of self-healing. Healthiness is the original condition of life. Modern medicine assumes sickness to be a natural phenomenon, with the essential nature of "Changes" neglected as a universal law for maintaining health. Dr. Sun, a renowned physician from the Tang Dynasty, was quoted as saying "Knowing Changes is the prerequisite of knowing medicine." Another saying holds that, "Every word and every sentence in the Book of Changes is an indicator of medicine." Much emphasis has been placed on the relationship between "Changes" and "medicine" in the past. This paper elaborates the relationship between nature and human health in order to provide a clear understanding of the nature of true health, described from the perspectives of medicine and "Changes", an evaluation of modern medical science and the oneness of body-mind-spirit, which is the reality of health. The human body is thus a reflection of the mind and spirit, while the mind and spirit is the "inner body". The body is a highly intelligent organism that truly reflects our inner world. Our inner world is also displayed through physical symptoms. As human suffering is caused by separation from our inner life, the only path to enjoying a healthy and joyful life is to achieve a oneness between our body-mind-spirit. Such is a universal law, which is called "Changes" or "Oneness".


Amyloid and the origin of life: self-replicating catalytic amyloids as prebiotic informational and protometabolic entities.
A crucial stage in the origin of life was the emergence of the first molecular entity that was able to replicate, transmit information, and evolve on the early Earth. The amyloid world hypothesis posits that in the pre-RNA era, information processing was based on catalytic amyloids. The self-assembly of short peptides into β-sheet amyloid conformers leads to extraordinary structural stability and novel multifunctionality that cannot be achieved by the corresponding nonaggregated peptides. The new functions include self-replication, catalytic activities, and information transfer. The environmentally sensitive template-assisted replication cycles generate a variety of amyloid polymorphs on which evolutive forces can act, and the fibrillar assemblies can serve as scaffolds for the amyloids themselves and for ribonucleotides proteins and lipids. The role of amyloid in the putative transition process from an amyloid world to an amyloid-RNA-protein world is not limited to scaffolding and protection: the interactions between amyloid, RNA, and protein are both complex and cooperative, and the amyloid assemblages can function as protometabolic entities catalyzing the formation of simple metabolite precursors. The emergence of a pristine amyloid-based in-put sensitive, chiroselective, and error correcting information-processing system, and the evolvement of mutualistic networks were, arguably, of essential importance in the dynamic processes that led to increased complexity, organization, compartmentalization, and, eventually, the origin of life.


Elevation of the soft palate in speech and swallowing in normal female participants and females with motor neuron disease: an innovative procedure for measuring palatal elevation.
The aim was to measure the angle of elevation of the soft palate during the separate conditions of swallowing of pudding, swallowing of liquid and during production of 'may/pay', using videofluroscopy and innovative computer software. We then determined whether soft palate elevation (as reflected in measures of an angle) differed in those separate conditions. Twelve female participants were studied, aged 40-70 years. The trends observed were that the highest soft palate elevation was displayed in the liquid-swallowing condition and the lowest soft palate elevation was displayed in the speech conditions. The results did not reach statistical significance, but the findings support the suggestions of Rubesin et al. (1987) and Fritzell (1969) that soft palate elevation is higher during swallowing than during speech. This same method was then used to measure soft palate elevation under the same three conditions in nine females, all aged > 40 years, who had motor neuron disease. Again, the highest soft palate elevation was displayed when swallowing liquid, but this time the lowest elevation occurred when swallowing pudding. Owing to small numbers, results did not reach statistical significance, but the trends may be important to replicate in a larger study. This pilot study developed a comprehensive and accurate method of measuring the soft palate elevation using digitization and computer software.


Association between PPAR-γ2 Pro12Ala polymorphism and obesity: a meta-analysis.
The peroxisome proliferator-activated receptor-γ2 (PPAR-γ2) gene has been reported in the pathogeny of obesity. However, the results have been inconsistent. The purpose of this meta-analysis was to acquire a more accurate assessment of the association between PPAR-γ2 Pro12Ala polymorphism and obesity. PubMed, Wan Fang (Chinese) databases, Chinese Biomedical Medical databases, and Chinese National Knowledge Infrastructure were searched to identify eligible studies. Finally, 25 studies (6491 cases and 8242 controls) were enrolled in this meta-analysis. The effect summary odds ratio (OR) with 95 % confidence interval (CI) was applied. Random-effects or fixed-effects model was performed based on the heterogeneity. STATA 12.0 was applied for this meta-analysis. The combined results showed that PPAR-γ Pro12Ala polymorphism was associated with the obesity risk (Ala vs. Pro: OR = 1.55, 95 % CI 1.34-1.80; Pro/Ala vs. Pro/Pro: OR = 1.54, 95 % CI 1.31-1.82; Ala/Ala + Pro/Ala vs. Pro/Pro: OR = 1.61, 95 % CI 1.36-1.90). Subgroup analysis by ethnicity showed that there were significant associations between PPAR-γ Pro12Ala polymorphism and obesity risk in Caucasians, Asians, and Mixed population. Subgroup analysis by obesity's cutoff points showed that the associations were found among the patients with the cutoff point of BMI ≥24 and BMI ≥30 but not among the patients with the cutoff point of BMI ≥95th percentile. These results suggested that PPAR-γ Pro12Ala polymorphism might be a risk factor for obesity susceptibility.


Decontamination of Mikania glomerata Leaves by Gamma Irradiation: Coumarin Determination by HPLC-DAD, Microbiological Control and Genotoxicological Studies.
Gamma irradiation as a decontaminating physical agent could be an important tool in the production chain of herbal medicines by improving the microbiological quality of raw materials and the safety of final products. This study was undertaken to investigate the genotoxic potential and eventual chemical modifications of a batch of Mikania glomerata raw material decontaminated by different doses of gamma irradiation (2.0, 3.5, and 5.0 kGy), using a cesium-137 source. DNA damage was assessed in vitro by agarose gel electrophoresis in regard to double-chain breaks of plasmid pUC 9.1 DNA and in vivo by micronucleus test in bone marrow cells of Wistar rats. Cytotoxicity in bone marrows was assessed by scoring polychromatic and normochromatic erythrocytes ratio. An HPLC-DAD method was adapted and validated for the enhancement of coumarin selectivity from the other matrix constituents. The microbial load was satisfactorily reduced, leading to sterilization at the highest dose. Genotoxic and cytotoxic effects were not increased in the in vitro and in vivo models. The concentration of coumarin and the chromatographic profiles of the hydroalcoholic plant extracts (ethanol 70% v/v) were not modified after such process. Therefore, this work suggests that gamma irradiation of M. glomerata raw material is suitable and safe for microbiological control purposes at the employed doses.


Defects in crystalline PVDF: a density functional theory-density functional tight binding study.
We present a comparative density functional theory (DFT) and density functional tight binding (DFTB) study of structures, energetics, vibrational properties as well as electronic structures of the four crystalline phases of polyvinylidene fluoride (PVDF) with different types of defects. For pure phases, the relative energies of PVDF strands (i.e. absent van der Walls bonding) agree well between DFT (using a GGA or a hybrid functional) and DFTB. For crystals, DFTB needs to be calibrated due to deficiencies in the treatment of vdW interactions. Defect formation energies were computed in large-scale DFTB simulations. For single chain vacancies, they are 0.41, 0.59, 0.08 and 0.40 eV per monomer removed in α, β, γ, and δ PVDF, respectively. The energy required to form double vacancies is 0.38, 0.52, 0.33 and 0.39 eV per monomer removed, respectively, i.e. the effect is nearly additive except in the γ phase. Interstitial defects were found to be unstable and convert into vacancies. The relatively high defect formation energies (vs. kT at room temperature) imply that phase purity is feasible in PVDF. Vibrational contributions affect the relative phase energies by up to 0.1 eV but do not significantly affect the relative phase stability.


Self-assembly and crystallization behavior of mesoporous, crystalline HfO2 thin films: a model system for the generation of mesostructured transition-metal oxides.
Mesoporous thin films of crystalline hafnium oxide were fabricated by evaporation-induced self-assembly in combination with sol-gel processing, followed by a suitable post heat-treatment procedure to initiate the crystallization. A novel type of block-copolymer template was used as structure-directing agent, which generated a distorted cubic arrangement of spherical mesopores, the size of which could be quantified by suitable techniques, such as ellipsometry-porosimetry, small-angle X-ray scattering, and atomic force microscopy. Detailed insights into the nature of the crystallization process of mesostructured hafnium oxide were obtained by temperature-dependent, in situ X-ray scattering experiments. These investigations revealed that crystallization takes place, within the confinement of the mesostructure, as a solid-solid transition from a dehydrated, amorphous form of hafnium oxide. The study suggests that one main benefit of the novel template results from the ability of the polymer to stabilize the mesostructure of amorphous hafnium oxide up to 400-450 degrees C.


Bilingual Phonological Awareness: Construct Validation in Grade 1 Spanish-Speaking English Learners.
This study investigated the dimensionality of bilingual phonological awareness (PA) in English and Spanish by replicating a kindergarten model in Grade 1, and presents alternatives to modeling clustered data. English and Spanish tasks were analyzed from previously collected samples totaling 1,586 first grade Spanish-speaking English learners. Four distinct approaches to confirmatory factor analysis (CFA) models were examined: (a) uncentered student-level data, (b) student-level data centered at the classroom means, (c) classroom-level data, and (d) multilevel CFA. Results indicated that while the multilevel CFA provided the most comprehensive view of the data, the multi-level student-level estimates were not appreciably different from estimates based on student-level data centered at the classroom means, and multi-level classroom-level estimates were comparable to estimates based on the analysis of classroom means. Importantly, English and Spanish PA were statistically separable at the student-level, but minimally distinct (r = .86) and slightly less correlated than what has been reported for kindergarten (r = .93). At the classroom level, the correlation was moderate (r = .51), and substantially reduced compared to kindergarten (r = .83). The distinction at the classroom-level between kindergarten and Grade 1 implies that instruction differentiates the abilities across languages at the classroom-level, but less so at the student-level.


Safety and antiviral response at 12 months of lopinavir/ritonavir therapy in human immunodeficiency virus-1-infected children experienced with three classes of antiretrovirals.
Many human immunodeficiency virus type 1 (HIV-1)-infected children have already failed treatment with 2 or even 3 classes of antiretrovirals. Coformulation of lopinavir with low dose ritonavir exhibits a potent antiretroviral effect. However, the data in heavily pretreated children are still scarce. This study evaluated the safety and effectiveness of combination therapy including lopinavir/ritonavir in children with prior exposure to all classes of oral antiretrovirals. This was an open label multicenter observational study, in which data were reviewed according to a standardized protocol. The study population included all HIV-1-infected children with virologic failure (HIV-1 RNA >5000 copies/mL) followed in 12 Spanish hospitals for >12 months, experienced with the 3 classes of oral antiretrovirals, in whom a lopinavir/ritonavir-containing regimen was started. By March 2003, 45 patients had been treated with lopinavir/ritonavir for a median of 18 months (range, 3-28). The median age at baseline was 9.7 years (range, 4.3-17.1). The median times of prior treatment were 88 months (range, 31-145) with nucleoside reverse transcription inhibitors and 42 months (range, 19-63) with protease inhibitors. Twenty-five patients were classified as Centers for Disease Control and Prevention clinical category C. Median values for absolute and percentage CD4 at baseline were 501 (range, 6-1512) and 19% (range, 0.5-49), respectively, and plasma HIV-RNA was 5.0 log10 copies/mL (range, 4.1-6.1). During follow-up, 11 (24%) children switched from liquid to solid formulation. At 48 weeks, the median values for absolute and percentage CD4 increased by 199 cells/microL and 3%, respectively, and median plasma viral load declined 1.75 log10 copies/mL. Forty-two percent of children achieved a plasma RNA of <400 copies/mL (intent to treat analysis). Baseline genotypic resistance was available in 40 children. Nonresponders had 7.0 +/- 1.6 protease inhibitor-associated mutations at baseline compared with 4.8 +/- 1.7 in children achieving virologic suppression (P = 0.06). Adverse events were described in 18 children. Three children permanently discontinued and 4 transiently withdrew lopinavir/ritonavir. At 12 months, there were mild but not significant increases in plasma cholesterol and triglycerides. Lopinavir/ritonavir when given as part of salvage regimen is well-tolerated, although switching to pills is frequently required. The regimen has a potent and durable antiretroviral activity in most heavily pretreated children, despite the presence of multiple mutations to all classes of oral antiretrovirals.


The role of telomere binding molecules for normal and abnormal hematopoiesis.
In order to maintain the homeostasis of the hematopoietic system, hematopoietic stem cells (HSCs) need to be maintained while slowly dividing over their lifetime. However, repeated cell divisions lead to the gradual accumulation of DNA damage and ultimately impair HSC function. Since telomeres are particularly fragile when subjected to replication stress, cells have several defense machinery to protect telomeres. Moreover, HSCs must protect their genome against possible DNA damage, while maintaining telomere length. A group of proteins called the shelterin complex are deeply involved in this two-way role, and it is highly resistant to the replication stress to which HSCs are subjected. Most shelterin-deficient experimental models suffer acute cytotoxicity and severe phenotypes, as each shelterin component is essential for telomere protection. The Tin2 point mutant mice show a dyskeratosis congenita (DC) like phenotype, and the Tpp1 deletion impairs the hematopoietic system. POT1/Pot1a is highly expressed in HSCs and contributes to the maintenance of the HSC pool during in vitro culture. Here, we discuss the role of shelterin molecules in HSC regulation and review current understanding of how these are regulated in the maintenance of the HSC pool and the development of hematological disorders.


The response of hepatic transcriptome to dietary cholesterol in Prague hereditary hypercholesterolemic (PHHC) rat.
To understand the pathogenesis of hypercholesterolemia in Prague hereditary hypercholesterolemic (PHHC) rat, we analyzed the response of hepatic transcriptome to dietary cholesterol in PHHC and control Wistar rats. Male PHHC and Wistar rats were fed chow (C), 5 % fat (palm kernel oil) (CF) or 1 % cholesterol + 5 % fat (CHOL) diet for three weeks. Hepatic transcriptome was analyzed using Affymetrix GeneChip arrays. No differences were found in the effect of both control diets (C and CF) on lipid metabolism and gene expression of 6500 genes. Therefore, these data were pooled for further analysis. Dietary cholesterol induced accumulation of cholesterol and triacylglycerols in the liver in both strains and hypercholesterolemia in PHHC rats. However, there were no differences in response of hepatic transcriptome to CHOL diet. On the other hand, several genes were found to be differently expressed between both strains independently of the diet. Two of those genes, Apof and Aldh1a7, were studied in more detail, and their role in pathogenesis of hypercholesterolemia in PHHC rats could not been corroborated. In conclusion, the hypercholesterolemia in PHHC rats is due to physiological response of hepatic transcriptome to dietary cholesterol in different genetic background.


Circular RNA-ZFR Inhibited Cell Proliferation and Promoted Apoptosis in Gastric Cancer by Sponging miR-130a/miR-107 and Modulating PTEN.
This study aimed to probe into the associations among circular RNA ZFR (circ-ZFR), miR-130a/miR-107, and PTEN, and to investigate the regulatory mechanism of circ-ZFR‒miR-130a/miR-107‒PTEN axis in gastric cancer (GC). GSE89143 microarray data used in the study were acquired from publicly available Gene Expression Omnibus database to identify differentially expressed circular RNAs inGC tissues. The expressions of circ-ZFR, miR-130a, miR-107, and PTEN were examined by real-time reverse transcription polymerase chain reaction, while PTEN protein expression was measured by western blot. The variation of GC cell proliferation and apoptosis was confirmed by cell counting kit-8 assay and flow cytometry analysis. The targeted relationships among circZFR, miR-130a/miR-107, and PTEN were predicted via bioinformatics analysis and demonstrated by dual-luciferase reporter assay and RNA immunoprecipitation assay. The impact of ZFR on gastric tumor was further verified in xenograft mice model experiment. Circ-ZFR and PTEN were low-expressed whereas miR-107 and miR-130a were highexpressed in GC tissues and cells. There existed targeted relationships and interactions between miR-130a/miR-107 and ZFR/PTEN. Circ-ZFR inhibited GC cell propagation, cell cycle and promoted apoptosis by sponging miR-107/miR-130a, while miR-107/miR-130a promoted GC cell propagation and impeded apoptosis through targeting PTEN. Circ-ZFR inhibited cell proliferation and facilitated apoptosis in GC by sponging miR-130a/miR-107 and modulating PTEN. Circ-ZFR curbed GC tumor growth and affected p53 protein expression in vivo. Circ-ZFR restrained GC cell proliferation, induced cell cycle arrest and promoted apoptosis by sponging miR-130a/miR-107 and regulating PTEN.


The volumetric relationship of white matter lesion and contrast-enhanced lesion in delayed radiation brain injury: an MRI-based study.
This study investigated the volumetric relationship of white matter lesion (WML) and contrast-enhanced lesion (CEL) in delayed radiation brain injury (RBI) during the course of evolution. MRI results in 45 patients with RBI after receiving radiation for nasopharyngeal carcinoma were analyzed. In total there were 75 lobes with RBI and 114 MRI examinations in this study. WML and CEL lesion volumes were measured. The lesion volume change of less than 5% or 0.25 cm(3) was regarded as being static. The average WML volume was 16.33 cm(3) (ranging 0.11 cm(3) to 102.83 cm(3)), and the average CEL volume was 3.15 cm(3) (ranging 0.03 cm(3) to 27.85 cm(3)). WML was larger than CEL in 164 measurements, and CEL was larger than WML in 10 measurements. In 64.3% follow-ups WML and CEL evolved in the same pattern; and in most follow-ups (93.8%) WML and CEL did not evolve in the opposite directions. A larger WML volume tended to have a larger CEL volume though this relationship was not linear. Evolution of WML and CEL tended to follow the same pattern. WML tended to be larger than CEL, and larger WML tended to be associated with larger CEL.


Neuroprotective strategies and the underlying molecular basis of cerebrovascular stroke.
Stroke is a leading cause of disability in the US. Although there has been significant progress in the area of medical and surgical thrombolytic technologies, neuroprotective agents to prevent secondary cerebral injury and to minimize disability remain limited. Only limited success has been reported in preclinical and clinical trials evaluating a variety of compounds. In this review, the authors discuss the most up-to-date information regarding the underlying molecular biology of stroke as well as strategies that aim to mitigate this complex signaling cascade. Results of historical research trials involving N-methyl-d-aspartate and α-amino-3-hydroxy-5-methyl-4-isoxazole propionate receptor antagonists, clomethiazole, antioxidants, citicoline, nitric oxide, and immune regulators have laid the groundwork for current progress. In addition, more recent studies involving therapeutic hypothermia, magnesium, albumin, glyburide, uric acid, and a variety of other treatments have provided more options. The use of neuroprotective agents in combination or with existing thrombolytic treatments may be one of many exciting areas of further development. Although past trials of neuroprotective agents in ischemic stroke have been limited, significant insights into mechanisms of stroke, animal models, and trial design have incrementally improved approaches for future therapies.


Fluorescence Detection of Deoxyadenosine in Cordyceps spp. by Indicator Displacement Assay.
A rapid, sensitive and reliable indicator displacement assay (IDA) for specific detection of 2'- and 3'-deoxyadenosine (2'-dAde and 3'-dAde), the latter is also known as cordycepin, was established. The formation of inclusion complex between protonated acridine orange (AOH+) and cucurbit[7]uril (CB7) resulted in the hypochromic shift of fluorescent emission from 530 nm to 512 nm. Addition of cordycepin to the highly fluorescent AOH+/CB7 complex resulted in a unique tripartite AOH+/CB7/dAde complex with diminished fluorescence, and such reduction in emission intensity serves as the basis for our novel sensing system. The detection limits were 11 and 82 μM for 2'- and 3'-deoxyadenosine, respectively. The proposed method also demonstrated high selectivity toward 2'- and 3'-deoxyadenosine, owing to the inability of other deoxynucleosides, nucleosides and nucleotides commonly found in Cordyceps spp. to displace the AOH+ from the AOH+/CB7 complex, which was confirmed by isothermal titration calorimetry (ITC), UV-Visible and proton nuclear magnetic resonance (1H-NMR) spectroscopy. Our method was successfully implemented in the analysis of cordycepin in commercially available Ophiocordyceps and Cordyceps supplements, providing a novel and effective tool for quality assessment of these precious fungi with several health benefits.


Effect of axial ligand on the electronic configuration, spin states, and reactivity of iron oxophlorin.
Iron-oxophlorin is an intermediate in heme degradation, and the nature of the axial ligand can alter the spin, electron distribution, and reactivity of the metal and the oxophlorin ring. The structure and reactivity of iron-oxophlorin in the presence of imidazole, pyridine, and t-butyl isocyanide as axial ligands was investigated using the B3LYP and OPBE methods with the 6-31+G* and 6-311+G** basis sets. OPBE/6-311+G** has shown that the doublet state of [(Py)(2)Fe(III)(PO)] (where pyridines are in perpendicular planes and PO is the oxophlorin trianion) is 3.45 and 5.27 kcal/mol more stable than the quartet and sextet states, respectively. The ground-state electronic configuration of the aforementioned complex is π(xz)(2) π(yz)(2) a(2u)(2) d(xy)(1) at low temperatures and changes to π(xz)(2) π(yz)(2) d(xy)(2) a(2u)(1) at high temperatures. This latter electronic configuration is consistently seen for the [(t-BuNC)(2)Fe(II)(PO(•))] complex (where PO(•) is the oxophlorin dianion radical). The complex [(Im)(2)Fe(III)(PO)] adopted the d(xy)(2) (π(xz) π(yz))(3) ground state and has low-lying quartet excited state which is readily populated when the temperature is increased.


Salvage of blood urea nitrogen in sheep is highly dependent on plasma urea concentration and the efficiency of capture within the digestive tract.
The aims of this study were 1) to determine whether transfer of blood urea to the gastrointestinal tract (GIT) or the efficiency of capture of urea N within the GIT is more limiting for urea N salvage, and 2) to establish the relationship between plasma urea concentration and recycling of urea N to the GIT. We used an i.v. urea infusion model in sheep to elevate the urea entry rate and plasma concentrations, thus avoiding direct manipulation of the rumen environment that otherwise occurs when feeding additional N. Four growing sheep (28.1 +/- 0.6 kg of BW) were fed a low-protein (6.8% CP, DM basis) diet and assigned to 4 rates of i.v. urea infusion (0, 3.8, 7.5, or 11.3 g of urea N/d; 10-d periods) in a balanced 4 x 4 Latin square design. Nitrogen retention (d 6 to 9), urea kinetics([(15)N2]urea infusion over 80 h), and plasma AA were determined. Urea infusion increased apparent total tract digestibility of N (29.9 to 41.3%) and DM (47.5 to 58.9%), and N retention (1.45 to 5.46 g/d). The plasma urea N entry rate increased (5.1 to 21.8 g/d) with urea infusion, as did the amount of urea N entering the GIT (4.1 to 13.2 g/d). Urea N transfer to the GIT increased with plasma urea concentration, but the increases were smaller at greater concentrations of plasma urea. Anabolic use of urea N within the GIT also increased with urea infusion (1.43 to 2.98 g/d; P = 0.003), but anabolic use as a proportion of GIT entry was low and decreased (35 to 22%; P = 0.003) with urea infusions. Consequently, much (44 to 67%) of the urea N transferred to the GIT returned to the liver for resynthesis of urea (1.8 to 9.2 g/d; P < 0.05). The present results suggest that transfer of blood urea to the GIT is 1) highly related to blood urea concentration, and 2) less limiting for N retention than is the efficiency of capture of recycled urea N by microbes within the GIT.


CITED2 functions as a molecular switch of cytokine-induced proliferation and quiescence.
Transforming growth factor-α (TGF-α)-induced proliferation and transforming growth factor-β (TGF-β)-mediated quiescence are intricately balanced in normal lung-tissue homeostasis but are deregulated during neoplastic progression of lung cancer. Here, we show that Cbp/p300-interacting transactivator with Glu/Asp-rich carboxy-terminal domain 2 (CITED2), a novel MYC-interacting transcriptional modulator, responds to TGF-α induction and TGF-β suppression to orchestrate cellular proliferation and quiescence, respectively. Upon TGF-α induction, CITED2 was induced by MYC and further modulated MYC-mediated transcription in a feed-forward manner. CITED2 recruited p300 to promote MYC-p300-mediated transactivation of E2F3, leading to increased G1/S cell cycle progression. Moreover, CITED2 inhibited cellular quiescence by enhancing MYC-mediated suppression of p21(CIP1). CITED2 interacted with histone deacetylase 1 (HDAC1) and potentiated MYC-HDAC1 complex formation. TGF-β stimulation provoked downregulation of CITED2, which abrogated MYC-HDAC1-mediated p21(CIP1) suppression, causing cellular quiescence. Ectopic CITED2 expression enhanced tumor growth in nude mice; furthermore, CITED2 knockdown caused tumor shrinkage and increased overall host mouse survival rates. Expression of CITED2/MYC/E2F3/p21(CIP1) signaling molecules was associated with poor prognosis of lung cancer patients. Thus, CITED2 functions as a molecular switch of TGF-α and TGF-β-induced growth control, and MYC-CITED2 signaling axis provides a new index for predicting clinical outcome.


Development of Polioencephalomyelitis in Cesarean-Derived Colostrum-Deprived Pigs Following Experimental Inoculation with Either Teschovirus A Serotype 2 or Serotype 11.
Teschovirus encephalomyelitis is a sporadic disease associated with Teschovirus A (PTV) serotype 1 and, less frequently, other serotypes. In recent years, the number of cases submitted to the Iowa State University Veterinary Diagnostic Laboratory with a history of posterior paresis has increased. Submission histories from various regions of the United States suggest a trend for clinical disease to persist in herds and affect a wider age-range of pigs than historically reported. Polioencephalitis and/or myelitis was consistently present and PTV was detected in affected neural tissue by PCR in a portion of cases. Sequencing from two clinical cases identified PTV-2 and PTV-11. To assess neuropathogenicity of these isolates, 5-week-old cesarean derived and colostrum-deprived pigs were assigned to three groups: negative control (n = 4), PTV-2-inoculated (n = 7), and PTV-11-inoculated (n = 7). Three PTV-2-inoculated pigs developed mild incoordination of the hind limbs, one of which progressed to posterior ataxia. While all PTV-11-inoculated pigs showed severe neurological signs consistent with Teschovirus encephalomyelitis, no evidences of neurological signs were observed in sham-inoculated animals. All PTV-2- and PTV-11-inoculated pigs had microscopic lesions consistent with Teschovirus encephalomyelitis. To our knowledge, this is the first description of PTV-11 and experimental study demonstrating the neuropathogenicity of PTV-11 in the United States.


Botulinum toxin complex increases paracellular permeability in intestinal epithelial cells via activation of p38 mitogen-activated protein kinase.
Clostridium botulinum produces a large toxin complex (L-TC) that increases paracellular permeability in intestinal epithelial cells by a mechanism that remains unclear. Here, we show that mitogen-activated protein kinases (MAPKs) are involved in this permeability increase. Paracellular permeability was measured by FITC-dextran flux through a monolayer of rat intestinal epithelial IEC-6 cells, and MAPK activation was estimated from western blots. L-TC of C. botulinum serotype D strain 4947 increased paracellular dextran flux and activated extracellular signal-regulated kinase (ERK), p38, but not c-Jun N-terminal kinase (JNK) in IEC-6 cells. The permeability increase induced by L-TC was abrogated by the p38 inhibitor SB203580. These results indicate that L-TC increases paracellular permeability by activating p38, but not JNK and ERK.


Mapping of disulfide bridges in antifreeze proteins from overwintering larvae of the beetle Dendroides canadensis.
Antifreeze proteins (AFPs) have been identified in certain high-latitude marine fish, insects and other terrestrial arthropods, and plants. Despite considerable structural variation, the mechanisms of their noncolligative antifreeze activity are probably quite similar. AFPs hydrogen bond onto the surface of potential seed ice crystals at preferred growth sites, thereby preventing growth of the crystals. AFPs from overwintering larvae of the beetle Dendroidescanadensis are among the most active AFPs. These 8.7-kDa proteins consist of seven 12- or 13-mer repeating units. Their most striking feature is the location of cysteines every six residues throughout their length. Consequently, identification of the disulfide linkages of these cysteines is essential to understanding the structure of these AFPs. This study demonstrated that all of the 16 Cys residues in the Dendroides AFPs are disulfide bridged. All of the seven 12- or 13-mer repeats have internal disulfide bridges, and in all but the first repeat the Cys residues at positions 1 and 7 of the repeats are linked. In repeat 1 the Cys at position 1 is linked to the Cys at position 10, rather than the Cys at position 7 as in the other repeats, and the Cys at position 7 of the first repeat is linked to a Cys at position 4 of the second repeat. The disulfide bridges probably function to position the hydrophilic side chains of serine and threonine residues so that they hydrogen bond with ice.


[4-nonylphenol combined with bisphenol A induces testicular autophagy and spermatogenic impairment by inhibiting the Akt/mTOR pathway in F1 male offspring rats].
To study the impacts of exposure of F0 generation rats to 4-nonylphenol (4-NP) and bisphenal A (BPA) on the autophagy of testicular cells and key gene expressions of the Akt/mTOR pathway in the F1 generation offspring rats. Using a 2×2 factorial design, F0 female and male rats were randomly assigned to receive intragastrically sigma vegetable oil (the control), BPA at 0.5 mg/kg, 4-NP at 5 mg/kg, and BPA+4-NP both at 0.5 mg/kg, respectively, qd alt for 30 days. Then the rats in each group were mated at a ratio of 1∶3. After pregnancy, the female rats continued the above intragastrical administration till delivery. The F1 generation male offspring rats were killed at 60 postnatal days, and their testes harvested for sperm count, observation of the morphological and autophagic changes of the testis, and determination of the relative mRNA expression levels of Akt, mTOR, 4EBP1 and p70S6K and protein expression levels of LC3-I and LC3-II. The sperm count of the F1 generation male offspring rats was markedly decreased in the BPA and BPA+4-NP groups compared with that in the control (P < 0.05) but no significant interactive effect was observed between BPA and 4-NP (P > 0.05). The ratio of LC3-II / LC3-I was remarkably increased in the 4-NP, BPA and BPA+4-NP groups in comparison with that in the control (P < 0.05), and a significant interactive effect was shown between BPA and 4-NP (P < 0.05). The cells in the seminiferous tubules of the rats in the 4-NP, BPA and BPA+4-NP groups were loosely arranged, with a decreased count of sperm and increased number of autophagic vacuoles. Significant down-regulation was observed in the relative mRNA expression of p70S6K in the 4-NP group, as well as in that of mTOR in the BPA group and Akt in the BPA+4-NP group (all P < 0.05). A remarkably up-regulated expression of 4EBP1 mRNA was found in all the three intervention groups (P < 0.05), with a significant interactive effect between BPA and 4-NP on the expressions of Akt and 4EBP1 mRNA (P < 0.05). Long-term exposure to low-concentration BPA and 4-NP impairs the testicular function of the F1 generation male offspring rats, which is closely related to the autophagy of testicular cells and changes of the Akt/mTOR pathway.


First Case of Enterovesical Fistula Caused by Ischaemic Enteritis.
We present the case of an enterovesical fistula (EVF) caused by ischemic enteritis. Ischemic enteritis is caused either by mesenteric macrovasculature occlusion or any pathophysiologic vasospasm of the microvasculature. In other words, ischemic enteritis (IE) occurs when the inflow of blood to the small intestines is reduced. The frequency of ischemic enteritis is less as compared to ischemic colitis because of the vast blood supply to the small intestine. It is the first case to be reported in the medical literature to date. EVF is a rare entity. It is a pathological connection between the bowel loops and the urinary bladder. EVF is a result of an underlying disease or injury. EVF is mostly caused by diverticular diseases, carcinoma colon, Crohn's, and inflammatory bowel disease, iatrogenic, appendicitis, carcinoma cervix, etc. Due to the formation of this abnormal connection, contents of the intestines, including the air, food content, etc., are usually found in the urine. Patients usually present with the complaint of irritative urinary tract symptoms and recurrent urinary tract infection (UTI). Surgical management is the mainstay of treatment although medical management is tried for those who cannot bear to undergo surgery.


The Burden of Cost in Bronchiolitis Obliterans Syndrome: Predictions for the Next Decade.
In health economics, costs can be divided into both direct and indirect categories. Direct costs tend to consist of medical costs, which are those directly attributed to health care interventions (e.g., hospitalizations, pharmaceuticals, devices), and non-medical direct costs such as monitoring and professional caregiving. Indirect costs tend to comprise those related to lost productivity due to illness (or treatment), burden on systems outside of the healthcare domain, and other costs that can sometimes outweigh the entire sum of direct healthcare costs. The most common life-threatening complication of lung and hematopoietic stem-cell transplantation (HSCT) is bronchiolitis obliterans syndrome (BOS). BOS is currently diagnosed as a 20% decline in the forced expiratory volume in one second (FEV1) from the best (baseline) post-transplantation value, and is a major cause of morbidity and mortality amongst lung and stem cell transplant patients. BOS affects half of all lung transplant patients within the first 5 years post-transplant, rising to the majority of patients (~80%) within the first decade following transplant. We estimated both direct and indirect costs for the first 10 years following BOS diagnosis, a viewpoint that highlights a tremendous imbalance between healthcare and non-healthcare costs. The lost workforce resulting from BOS-related infirmity will cost society more than $3.7 Billion over the next decade, a figure that is more than double the estimated 10-year cost of treating BOS ($1.4B), including diagnostics, immunosuppressives, and additional complications. As such, BOS is estimated to present a burden of cost that must be evaluated in a new light to include the wider societal perspective.


Sum-over-states calculation of the specific rotations of some substituted oxiranes, chloropropionitrile, ethane, and norbornenone.
A sum-over-states approach has been applied to the calculation of the specific rotations of several substituted oxiranes, 2-chloropropionitrile, and 30 degrees-rotated ethane. In each case, the first few excited states proved to have only a relatively small effect on the calculated specific rotation. It was necessary to use a very large number of excited states in order to achieve convergence with the results of the more direct linear response method. However, the latter does not give information on which excited states are important in determining the specific rotation. Norbornenone is unique in that its greatly enhanced specific rotation as compared to norbornanone is associated with the low-energy n-pi* transition. The C=C bond orbitals interact with the C=O in the LUMO, and a density difference plot for going from the ground state to the first excited state clearly shows the perturbation of the C=C.


Downregulation of transient receptor potential M6 channels as a cause of hypermagnesiuric hypomagnesemia in obese type 2 diabetic rats.
We assessed the expression profile of Mg(2+)-transporting molecules in obese diabetic rats as a cause of hypermagnesiuric hypomagnesemia, which is involved in the development of insulin resistance, hypertension, and coronary diseases. Kidneys were obtained from male Otsuka Long-Evans Tokushima fatty (OLETF) and Long-Evans Tokushima Otsuka (LETO) obese diabetic rats at the ages of 16, 24, and 34 wk. Expression profiles were studied by real-time PCR and immunohistochemistry together with measurements of urine Mg(2+) excretion. Urine Mg(2+) excretion was increased in 24-wk-old OLETF rats and hypomagnesemia was apparent in 34-wk-old OLETF rats but not in LETO rats (urine Mg(2+) excretion: 0.16 ± 0.01 μg·min(-1)·g body wt(-1) in 24-wk-old LETO rats and 0.28 ± 0.01 μg·min(-1)·g body wt(-1) in 24-wk-old OLETF rats). Gene expression of transient receptor potential (TRP)M6 was downregulated (85.5 ± 5.6% in 34-wk-old LETO rats and 63.0 ± 3.5% in 34-wk-old OLETF rats) concomitant with Na(+)-Cl(-) cotransporter downregulation, whereas the expression of claudin-16 in tight junctions of the thick ascending limb of Henle was not different. The results of the semiquantitative analysis of immunohistochemistry were consistent with these findings (TRPM6: 0.49 ± 0.04% in 16-wk-old LETO rats, 0.10 ± 0.01% in 16-wk-old OLETF rats, 0.52 ± 0.03% in 24-wk-old LETO rats, 0.10 ± 0.01% in 24-wk-old OLETF rats, 0.48 ± 0.02% in 34-wk-old LETO rats, and 0.12 ± 0.02% in 34-wk-old OLETF rats). Gene expression of fibrosis-related proinflammatory cytokines as well as histological changes showed that the hypermagnesiuria-related molecular changes and tubulointerstitial nephropathy developed independently. TRPM6, located principally in distal convoluted tubules, appears to be a susceptible molecule that causes hypermagnesiuric hypomagnesemia as a tubulointerstitial nephropathy-independent altered tubular function in diabetic nephropathy.


Human Adipose-Derived Mesenchymal Stem Cells Respond to Short-Term Hypoxia by Secreting Factors Beneficial for Human Islets In Vitro and Potentiate Antidiabetic Effect In Vivo.
Adipose-derived mesenchymal stem cells (ASCs) release factors beneficial for islets in vitro and protect against hyperglycemia in rodent models of diabetes. Oxygen tension has been shown to induce metabolic changes and alter ASCs' release of soluble factors. The effects of hypoxia on the antidiabetic properties of ASCs have not been explored. To investigate this, we incubated human ASCs for 48 h in 21% (normoxia) or 1% O2 (hypoxia) and compared viability, cell growth, surface markers, differentiation capability, and soluble factors in the conditioned media (CM). Human islets were exposed to CM from ASCs incubated in either normoxia or hypoxia, and islet function and apoptosis after culture with or without proinflammatory cytokines were measured. To test hypoxic preconditioned ASCs' islet protective effects in vivo, ASCs were incubated for 48 h in normoxia or hypoxia before being injected into Balb/c Rag 1-/- immunodeficient mice with streptozotocin-induced insulitis. Progression of diabetes and insulin content of pancreas were measured. We found that incubation in hypoxia was well tolerated by ASCs and that levels of VEGF-A, FGF-2, and bNGF were elevated in CM from ASCs incubated in hypoxia compared to normoxia, while levels of HGF, IL-8, and CXCL1 were reduced. CM from ASCs incubated in hypoxia significantly improved human islet function and reduced apoptosis after culture, and reduced cytokine-induced apoptosis. In our mouse model, pancreas insulin content was higher in both groups receiving ASCs compared to control, but the mice receiving preconditioned ASCs had lower random and fasting blood glucose, as well as improved oral glucose tolerance compared to untreated mice. In conclusion, our in vitro results indicate that the islet protective potential of ASCs improves in hypoxia, and we give insight into factors involved in this. Finally we show that hypoxic preconditioning potentiates ASCs' antidiabetic effect in vivo.


Inactivation and stabilization of stabilisins in neat organic solvents.
The stability of the serine proteases from Bacillus amyloliquefaciens (subtillisin BPN') and Bacillus licheniformis (subtilisin Carlsberg) was investigated in various anhydrous solvents at 45 degrees C. The half-life of subtilisin BPN' in dimethyl-formamide dramatically depends on the pH of the aqueous solutions from which the enzyme was lyophilized, increasing from 48 min to 20 h when the pH is raised from 6.0 to 7.9. Both subtilisins exhibited substantial inactivation during multihour incubations in tert-amyl alcohol and acetonitrile when enzymatic activities were also measured in these solvents; however, when the enzymes were assayed in water instead, hardly any loss of activity was detected. This surprising difference appears to stem from the partitioning of the bound water essential for catalytic activity from the enzymes into the solvents. When assayed in organic solvents, this time-dependent stripping of water results in decay of enzymatic activity; however, when assayed in water, where the dehydrated subtilisins can undergo rehydration thereby recovering catalytic activity, little inactivation is observed. In agreement with this hypothesis, the addition of small quantities of water tert-amyl alcohol stabilized the subtilisins in it even when enzymatic activity was measured in the nonaqueous solvent. Ester substrates (vinyl butyrate and trichloroethyl butyrate) greatly enhanced the stability of both subtilisins in organic solvents possibly because of the formation of the acyl-enzymes.


Bulimia: an entity in search of definition.
A review of research literature that addresses personality correlates of bulimia revealed a major problem which affects that research and its interpretation: Criteria used to define this disorder have not been consistent across investigators. A related issue is that this disorder has been described by a variety of names, such as dietary chaos syndrome, binge-eating syndrome, bulimia nervosa, etc. The second major problem that contributes to the uncertainty about the personality of bulimics is methodology. Among these have been the inclusion of individuals with a history of anorexia nervosa, the use of researcher-specific self-reports and questionnaires (primarily within a narrow population), and a lack of consistency in the testing measures used to assess personality. This very basic issue needs to be resolved if the findings in bulimia research are to be cumulative and generalizable across studies. Suggestions are offered as to how this resolution should occur.


NIH/3T3 Fibroblasts Selectively Activate T Cells Specific for Posttranslationally Modified Collagen Type II.
It has been shown that synovial fibroblasts (SF) play a key role in the initiation of inflammation and joint destruction, leading to arthritis progression. Fibroblasts may express major histocompatibility complex class II region (MHCII) molecules, and thus, they could be able to process and present antigens to immunocompetent cells. Here we examine whether different types of fibroblasts (synovial, dermal, and thymic murine fibroblasts, destructive LS48 fibroblasts, and noninvasive NIH/3T3 fibroblasts) may be involved in the initiation of rheumatoid arthritis (RA) pathogenesis and can process and present type II collagen (COL2)-an autoantigen associated with RA. Using a panel of MHCII/Aq-restricted T-cell hybridoma lines that specifically recognize an immunodominant COL2 epitope (COL2259-273), we found that NIH/3T3 fibroblasts activate several T-cell clones that recognize the posttranslationally glycosylated or hydroxylated COL2259-273 epitope. The HCQ.3 hybridoma, which is specific for the glycosylated immunodominant COL2 epitope 259-273 (Gal264), showed the strongest response. Interestingly, NIH/3T3 cells, but not destructive LS48 fibroblasts, synovial, dermal, or thymic fibroblasts, were able to stimulate the HCQ.3 hybridoma and other COL2-specific T-cell hybridomas. Our experiments revealed that NIH/3T3 fibroblasts are able to activate COL2-specific T-cell hybridomas even in the absence of COL2 or a posttranslationally modified COL2 peptide. The mechanism of this unusual activation is contact-dependent and involves the T-cell receptor (TCR) complex.


Cross-Talk Between m6A- and m5C-Related lncRNAs to Construct a Novel Signature and Predict the Immune Landscape of Colorectal Cancer Patients.
N6-methyladenosine (m6A) and 5-methylcytosine (m5C) can modify long non-coding RNAs (lncRNAs), thereby affecting tumorigenesis and tumor progression. However, there is a lack of knowledge regarding the potential roles and cross-talk of m6A- and m5C-related lncRNAs in the tumor microenvironment (TME) and their effect on prognosis. We systematically evaluated the expression patterns of m6A- and m5C-related lncRNAs in 1358 colorectal cancer (CRC) samples from four datasets. Consensus clustering was conducted to identify molecular subtypes of CRC, and the clinical significance, TME, tumor-infiltrating immune cells (TIICs), and immune checkpoints in the different molecular subtypes were analyzed. Finally, we established a m6A- and m5C-related lncRNA signature and a prognostic nomogram. We identified 141 m6A- and m5C-related lncRNAs by co-expression analysis, among which 23 lncRNAs were significantly associated with the overall survival (OS) of CRC patients. Two distinct molecular subtypes (cluster A and cluster B) were identified, and these two distinct molecular subtypes could predict clinicopathological features, prognosis, TME stromal activity, TIICs, immune checkpoints. Next, a m6A- and m5C-related lncRNA signature for predicting OS was constructed, and its predictive capability in CRC patients was validated. We then constructed a highly accurate nomogram for improving the clinical applicability of the signature. Analyses of clinicopathological features, prognosis, TIICs, cancer stem cell (CSC), and drug response revealed significant differences between two risk groups. In addition, we found that patients with a low-risk score exhibited enhanced response to anti-PD-1/L1 immunotherapy. Functional enrichment analysis showed that these lncRNAs related to the high-risk group were involved in the development and progression of CRC. We conducted a comprehensive analysis of m6A- and m5C-related lncRNAs in CRC and revealed their potential functions in predicting tumor-immune-stromal microenvironment, clinicopathological features, and prognosis, and determined their role in immunotherapy. These findings may improve our understanding of the cross-talk between m6A- and m5C-related lncRNAs in CRC and pave a new road for prognosis assessment and more effective immunotherapy strategies.


Recent advances in the management of priapism.
Priapism is an uncommon urological emergency that can lead to permanent impotence if prompt presentation and medical intervention is not performed. It is a breakdown of the usual physiological mechanisms controlling penile tumescence and detumescence, leading to a prolonged penile erection (>4 hours) that is unrelated to sexual stimulation. Currently, there are three accepted subtypes: ischaemic, non-ischaemic, and stuttering priapism, which is also known as recurrent ischemic priapism. The aim of treatment is the immediate resolution of the painful erection and the preservation of cavernosal smooth muscle function in order to prevent cavernosal fibrosis, which can lead to penile shortening and permanent erectile dysfunction.


Minimally invasive repair of pectus excavatum in adult patients.
Since 1996, the technique for minimally invasive repair of pectus excavatum (MIRPE) has gained increasing acceptance among pediatric patients. However, the feasibility of the operation and outcomes have not yet been evaluated in adult patients. This study was a retrospective analysis of the author's experience combined with a survey of members of the American Pediatric Surgical Association in treating adult patients with MIRPE. Thirty adults (age range, 18-32 years; mean, 23 years; 75% men) with severe pectus excavatum (chest index > 3.2) were treated with MIRPE. The main indication for surgery was cosmetic (80%). One 32-year-old female patient underwent simultaneous MIRPE and breast augmentation. In 60 per cent of cases, the operative time was 1 to 2 hours. Thoracic epidural was successfully used for postoperative pain management in 90 per cent of patients. Two pectus bars were necessary in 16 per cent of cases, and bilateral stabilizers were used in 53 per cent of patients. Complications included seroma (10%), bar displacement (6%), pneumothorax requiring tube thoracostomy (6%), superficial wound infection (3%), and stabilizer bar fracture (3%). Two patients required conversion to modified Ravitch repair. Patient satisfaction was rated as excellent (50%), good (36%), and fair (14%). Less than 50 per cent of patients achieved 100 per cent correction of their deformity. MIRPE can be used safely for repair of pectus excavatum in adult patients. The complication rate appears to be similar to previously reported series of pediatric patients. Although adult patients may have residual asymmetry of the chest postrepair, overall satisfaction with the repair was very good or excellent in 86 per cent of patients.