Medullary and spinal cord projections from cardiovascular responsive sites in the rostral ventromedial medulla.
The rostral ventromedial medulla (RVMM) is a sympathoexcitatory area. However, little is known about its efferent projections. In this study, biotinylated dextran amine (BDA) or Phaseolus vulgaris leucoagglutinin (PHA-L) were used to investigate the medullary and spinal cord projections from pressor sites in RVMM. Initially, RVMM was systematically explored in urethane-anesthetized rats using microinjection of L-glutamate for sites that elicited increases in arterial pressure. A pressor area was identified that included the rostral magnocellular reticular and rostral lateral paragigantocellular reticular nuclei. In the second series of experiments, BDA or PHA-L was iontophoretically injected into RVMM pressor sites. Anterograde labeling was observed throughout the brainstem and spinal cord, bilaterally, but with an ipsilateral predominance. Dense labeling was observed within the nucleus of the solitary tract (NTS); the greatest density of labeling was observed in the caudal dorsolateral, medial, and ventrolateral subnuclei. Additionally, light to moderately dense labeling was found within the nucleus substantia gelatinosus and commissural nucleus. In the nucleus ambiguus/ventrolateral medullary (Amb/VLM) region, the density of labeling was greatest in caudal regions. Within Amb, most of the labeling was localized to its external formation. Anterograde labeling was also found throughout the spinal cord. In the thoracolumbar segments, dense axonal labeling was observed within the dorsolateral funiculus. These labeled axons innervated the intermediolateral nucleus and the central autonomic area. Taken together, these data suggest that RVMM neurons elicit increases in sympathetic activity by likely providing a direct excitatory input to spinal sympathetic preganglionic neurons, and by a direct inhibitory input to medullary cardioinhibitory and depressor areas.


A peripheral pacemaker drives the circadian rhythm of synaptic boutons in Drosophila independently of synaptic activity.
Circadian rhythms in the morphology of neurons have been demonstrated in the fly Drosophila melanogaster. One such rhythm is characterized by changes in the size of synaptic boutons of an identified flight motor neuron, with larger boutons during the day compared with those at night. A more detailed temporal resolution of this rhythm shows here that boutons grow at a time of increased locomotor activity during the morning but become gradually smaller during the day and second period of increased locomotor activity in the evening. We have experimentally manipulated the synaptic activity of the fly during short periods of the day to investigate whether changes in bouton size might be a consequence of the different levels of synaptic activity associated with the locomotion rhythm of the fly. In the late night and early morning, when the flies normally have an intense period of locomotion, the boutons grow independently of whether the flies are active or completely paralyzed. Bouton size is not affected by sleep-deprivation during the early night. The cycle in bouton size persists for 2 days even in decapitated flies, which do not move, reinforcing the notion that it is largely independent of synaptic activity, and showing that a pacemaker other than the main biological clock can drive it.


Multi-targeted antiangiogenic tyrosine kinase inhibitors in advanced non-small cell lung cancer: meta-analyses of 20 randomized controlled trials and subgroup analyses.
Multi-targeted antiangiogenic tyrosine kinase inhibitors (MATKIs) have been studied in many randomized controlled trials (RCTs) for treatment of advanced non-small cell lung cancer (NSCLC). We seek to summarize the most up-to-date evidences and perform a timely meta-analysis. Electronic databases were searched for eligible studies. We defined the experimental arm as MATKI-containing group and the control arm as MATKI-free group. The extracted data on objective response rates (ORR), disease control rates (DCR), progression-free survival (PFS) and overall survival (OS) were pooled. Subgroup and sensitivity analyses were conducted. Twenty phase II/III RCTs that involved a total of 10834 participants were included. Overall, MATKI-containing group was associated with significant superior ORR (OR 1.29, 95% CI 1.08 to 1.55, P = 0.006) and prolonged PFS (HR 0.83, 0.78 to 0.90, P = 0.005) compared to the MATKI-free group. However, no significant improvements in DCR (OR 1.08, 1.00 to 1.17, P = 0.054) or OS (HR 0.97, 0.93 to 1.01, P = 0.106) were observed. Subgroup analyses showed that the benefits were predominantly presented in pooled results of studies enrolling previously-treated patients, studies not limiting to enroll non-squamous NSCLC, and studies using MATKIs in combination with the control regimens as experimental therapies. This up-to-date meta-analysis showed that MATKIs did increase ORR and prolong PFS, with no significant improvement in DCR and OS. The advantages of MATKIs were most prominent in patients who received a MATKI in combination with standard treatments and in patients who had previously experienced chemotherapy. We suggest further discussion as to the inclusion criteria of future studies on MATKIs regarding histology.


Lasso Proteins-Unifying Cysteine Knots and Miniproteins.
Complex lasso proteins are a recently identified class of biological compounds that are present in considerable fraction of proteins with disulfide bridges. In this work, we look at complex lasso proteins as a generalization of well-known cysteine knots and miniproteins (lasso peptides). In particular, we show that complex lasso proteins with the same crucial topological features-cysteine knots and lasso peptides-are antimicrobial proteins, which suggests that they act as a molecular plug. Based on an analysis of the stability of the lasso piercing residue, we also introduce a method to determine which lasso motif is potentially functional. Using this method, we show that the lasso motif in antimicrobial proteins, as well in that in cytokines, is functionally relevant. We also study the evolution of lasso motifs, their conservation, and the usefulness of the lasso fingerprint, which extracts all topologically non-triviality concerning covalent loops. The work is completed by the presentation of extensive statistics on complex lasso proteins to analyze, in particular, the strange propensity for "negative" piercings. We also identify 21 previously unknown complex lasso proteins with an ester and a thioester bridge.


Unbiased bootstrap error estimation for linear discriminant analysis.
Convex bootstrap error estimation is a popular tool for classifier error estimation in gene expression studies. A basic question is how to determine the weight for the convex combination between the basic bootstrap estimator and the resubstitution estimator such that the resulting estimator is unbiased at finite sample sizes. The well-known 0.632 bootstrap error estimator uses asymptotic arguments to propose a fixed 0.632 weight, whereas the more recent 0.632+ bootstrap error estimator attempts to set the weight adaptively. In this paper, we study the finite sample problem in the case of linear discriminant analysis under Gaussian populations. We derive exact expressions for the weight that guarantee unbiasedness of the convex bootstrap error estimator in the univariate and multivariate cases, without making asymptotic simplifications. Using exact computation in the univariate case and an accurate approximation in the multivariate case, we obtain the required weight and show that it can deviate significantly from the constant 0.632 weight, depending on the sample size and Bayes error for the problem. The methodology is illustrated by application on data from a well-known cancer classification study.


Extrarenal malignant rhabdoid tumour of the heel--a case report.
Malignant rhabdoid tumour is a rare soft-tissue neoplasm that occurs in children and young adults. Cases have involved numerous extrarenal sites, including the thymus, liver, paravertebral region, central nervous system, heart, prostate, pelvis, chest wall, extremities, as well as soft tissues. Given their highly aggressive nature, early diagnosis of rhabdoid tumours is essential; indeed, survival times after initial presentation are typically in the range of months rather than years. The low incidence of rhabdoid tumours and confusion between this type of tumour and other tumours has precluded the collection of adequate data on its clinical presentation and there is no specific therapeutic protocol. Owing to the rarity of this condition and the difficulties encountered in its diagnosis and treatment, we report the case of a malignant tumour with rhabdoid-like features in the foot that confirms the local aggressiveness and the high metastatic potential of this type of tumour.


Quality by design (QbD) approach for design and development of drug-device combination products: a case study on flunisolide nasal spray.
The objective of the present study was to design and develop drug-device combination product in particular flunisolide nasal spray (FNS) using quality by design (QbD) approach. Quality target product profile (QTPP) of FNS was defined and critical quality attributes (CQAs), i.e. viscosity (cp) (Y1) and D50 droplet size distribution (DSD) (μm) (Y2) were identified. Potential risk factors were identified using a fish bone diagram and failure mode effect analysis (FMEA) tools. Plackett-Burman and Box-Behnken designs were used for screening the significant factors and optimizing the variables range, respectively. It was observed that viscosity (cp) (Y1) was significantly impacted by formulation variables X1: propylene glycol (PG) (%) and X2: polyethylene glycol (PEG) 3350 (%), while D50 DSD (μm) (Y2) was significantly impacted by formulation variables X1: PG (%), X2: PEG 3350 (%) and device variable X8: delivery volume (μl). A design space plot within which the CQAs remained unchanged was established at laboratory scale. In conclusion, this study demonstrated how QbD based development approach can be applied to the development of drug-device combination products with enhanced understanding of the impact of formulation, process and device variables on CQAs of drug-device combination products.


[Genetics and epidemiology of retinoblastoma].
Recent studies of genetic epidemiology of retinoblastoma are reviewed, and major conclusions drawn are as follows: Retinoblastoma occurs in heritable or nonheritable form. The primary genetic change in the heritable form is a point mutation or deletion at a locus on 13 q 14. For the genesis of a tumor in either form, the loss or inactivation of both alleles at this locus is a prerequisite, although additional events may be needed for its development. Information about environmental risk factors for the occurrence of heritable or nonheritable retinoblastoma is still meager. Although viral etiology for nonheritable tumor cannot be excluded, an extensive epidemiologic study revealed no seasonal variation in the births of 675 patients with sporadic unilateral cases, a great majority of which may be regarded as due to somatic mutation. Moreover, there was no paternal age effect at all on the occurrence of 225 sporadic bilateral cases. Paternal exposure to ionizing radiation or chemical mutagens, which should have an accumulated effect with advancing age, does not seem to play a major role in the production of germinal mutation at the Rb locus. Family studies show that host resistance genes at other loci can modify the process of tumor development when the primary genetic change is already present in all the target cells. Unaffected gene carriers may be regarded as inherently resistant to tumor formation, whereas persons who present early onset of bilateral tumors are the most susceptible. In certain families, however, non-expression in the carriers may be due to chromosomal rearrangement in a balanced state.


Is disturbed intracortical excitability a stable trait of chronic insomnia? A study using transcranial magnetic stimulation before and after multimodal sleep therapy.
Chronic insomnia is a poorly understood disorder. Risk factors for developing chronic insomnia are largely unknown, yet disturbances in brain indexes of arousal seem to accompany the disorder. We here investigate whether insomnia patients and control participants differ with respect to brain responses to direct stimulation, i.e., cortical excitability. Transcranial magnetic stimulation (TMS) offers a method to directly investigate the excitability level of the human cerebral cortex in psychiatric and neurological disease. We investigated cortical excitability in 16 insomnia patients and 14 carefully matched control participants using absolute and relative amplitudes of motor evoked potentials in response to single- and paired-pulse stimulation using TMS. Nonmedicated insomnia patients showed, first, an exaggerated absolute response to both suprathreshold single- and paired-pulse stimulation compared with control participants and second, a reduced relative response to paired-pulse stimulation at long interpulse intervals (i.e., a reduced intracortical facilitation). The abnormal excitability persisted despite sleep therapy that effectively improved sleep quality as well as behavioral and neuroimaging indexes of brain function. The results suggest that a subtly disturbed intracortical excitability characterizes patients with chronic insomnia: a relatively reduced intracortical facilitation in the context of a globally increased absolute excitability. The findings do not resemble TMS findings after sleep deprivation or in sleep apnea and thus seem specific to insomnia. They may offer diagnostic value and implications for assessment of risk to develop this common and disabling disorder.


Number of concomitant drugs with thrombocytopenic adverse effects and the extent inflammatory response resolution are risk factors for thrombocytopenia in patients treated with linezolid for more than 14 days.
Prolonged treatment with linezolid (LZD) is known to cause thrombocytopenia. However, some patients do not develop thrombocytopenia despite long-term administration of LZD. To determine the risk factors for LZD-associated thrombocytopenia in patients undergoing long-term LZD therapy, we conducted a retrospective cohort study among 212 patients receiving LZD treatment between December 2011 and June 2014 at a tertiary referral university hospital in Nagoya, Japan. Of the 217 patients who received LZD, 37 were treated with LZD for more than 14 days and were enrolled in the study. We compared data on demographic characteristics, underlying disease, microbiology, concomitant drugs, and laboratory tests between the thrombocytopenia group and the non-thrombocytopenia group. Thrombocytopenia was defined as having a platelet count < 100 × 103/μL or a ≥ 50% reduction in platelet count compared to baseline. Among the 37 patients who received LZD for more than 14 days, 17 (45.9%) developed thrombocytopenia. Multivariate logistic regression revealed that both the number of concomitant drugs with thrombocytopenic adverse effects (DTADE) (OR = 1.690; 95% CI = 1.037-2.754; P = 0.035) and a small decrease in the level of C-reactive protein (CRP) 14 days post-administration (OR = 0.965; 95% CI = 0.939-0.993; P = 0.013) were associated with thrombocytopenia during long-term LZD therapy. Therefore, the number of concomitant DTADE and a small decrease in CRP on the 14th day of treatment were key factors for the appearance of LZD-associated thrombocytopenia in patients with long-term LZD therapy. Our findings may be useful for preventing thrombocytopenia in patients treated with LZD for longer than 14 days.


Memories are made of this (perhaps): a review of serotonin 5-HT(6) receptor ligands and their biological functions.
The possible role of 5-HT(6) receptor antagonists in the treatment of learning and memory disorders has stimulated significant recent work in this area. The first selective antagonists of this receptor were identified by Roche (Ro 04-6790 and Ro 63-0563) and SmithKline Beecham (SB-271046), although they only had poor to modest brain penetration, respectively. Recently, several structurally different series of selective antagonists have been reported. Glennon s group and Merck Sharp & Dohme have discovered N,N-dimethyl-1-benzenesulfonyl-5-methoxytrypt amine as a reasonably selective, high affinity antagonist, while Allelix have gone on to find that a 6-bicyclopiperazinyl-1-naphthylsulfonylindole had improved affinity and selectivity. Roche have reported subsequently on more lipophilic analogs of Ro 04-6790 that appear to penetrate the brain better. Reversing the sulfonamide linkage of SB-271046 led to a new series of compounds, producing SB-357134, which also had increased CNS penetration. A series of selective partial agonists containing a 4-piperazinylquinoline system has also been described. Recent studies in the Morris water maze with both Ro 04-6790 and SB-271046 have concluded that 5-HT(6) receptor antagonists improved retention performance, although these results are open to interpretation. Other behavioural studies have also implicated a role for 5-HT(6) in cognition enhancement and this has been supported by in vivo microdialysis studies that showed SB-271046 produced an increase in extracellular glutamate levels in the frontal cortex. However, we have been unable to replicate these effects with either SB-271046 or Ro 04-6790, and clearly further work is required before we can be certain of the functional role of this receptor.


Effects of catechins on the mouse tumor cell adhesion to fibronectin.
We studied the effects of 5 kinds of catechins on the adhesion of mouse lung carcinoma 3LL and melanoma B16F10 cells to the fibronectin substratum. (-)-Epicatechin gallate and (-)-epigallocatechin gallate were active in inhibiting the 3LL cell adhesion, while (+)-catechin, (-)-epicatechin, and (-)-epigallocatechin were inactive. Gallate-containing catechins also impaired adhesion and/or spreading of B16F10 cells.


Assessing risk of a prolonged QT interval-a survey of emergency physicians.
Although QT prolongation is associated with an increased risk of torsades de pointes (TdP), it is unclear how clinicians determine risk in individual patients with prolonged QT. To investigate physicians' interpretation of electrocardiogram (ECG) values in patients with a prolonged QT in reference to risk of TdP. A survey was sent to Australasian emergency physicians (EPs) to investigate interpretation of ECG data in risk assessment for TdP. The survey contained three sections: demographic information, questions on heart rate correction and six sets of ECG data which the clinician ranked from low to high risk. Risk analysis for ECG values was performed by producing histograms of the distribution of responses for each of the six sets of ECG parameters. These distributions were compared to predicted distributions based on Bazett's corrected QT > 500 ms and the QT nomogram. The QT nomogram is a recently developed method for assessing whether QT-HR pairs are associated with increased risk of TdP by plotting them to determine if they are above an at risk line-the nomogram. Of 720 surveys sent out, 249 were returned (35%). A heart rate correction was used by 90% of respondents and the median "at risk" QTc judged by EPs was 450 ms [interquartile range (IQR): 440-500 ms]. Respondents were divided as to whether bradycardia increased the risk of TdP, with equal numbers responding "no change" and "more caution". In four of the six sets of ECG parameters, EPs had a similar risk distribution to that predicted by Bazett. For one point predicted to be high risk by the QT nomogram, there was a uniform (undecided) risk distribution by EPs. EPs mainly relied on Bazett's correction as their method of TdP risk assessment, which may be problematic for bradycardic patients.


Polyamines and polyamine-metabolizing enzymes in schizophrenia: Current knowledge and concepts of therapy.
Polyamines play preeminent roles in a variety of cellular functions in the central nervous system and other organs. A large body of evidence suggests that the polyamine pathway is prominently involved in the etiology and pathology of schizophrenia. Alterations in the expression and activity of polyamine metabolizing enzymes, as well as changes in the levels of the individual polyamines, their precursors and derivatives, have been measured in schizophrenia and animal models of the disease. Additionally, neuroleptic treatment has been shown to influence polyamine concentrations in brain and blood of individuals with schizophrenia. Thus, the polyamine system may appear to be a promising target for neuropharmacological treatment of schizophrenia. However, for a number of practical reasons there is currently only limited hope for a polyamine-based schizophrenia therapy.


Anti-IgE efficacy in murine asthma models is dependent on the method of allergen sensitization.
Murine models used to delineate mechanisms and key mediators of asthma have yielded conflicting results and suggest that the dominant mechanism and mediators required for disease induction differ depending on the model and method of allergen sensitization used. The goal of this study was to determine whether the mode of allergen sensitization influenced the role that IgE had in allergen-induced pulmonary eosinophilic inflammation. Mice were exposed to dust mite extract in 2 models of allergic inflammation that differed in the method of sensitization. We compared sensitization by aerosol exposure with and without concomitant human respiratory syncytial virus infection with sensitization by means of systemic (intraperitoneal) exposure with adjuvant. After sensitization, animals were similarly challenged with aerosolized allergen. Animals were treated with anti-IgE mAb to deplete IgE and to determine its role in the induction of allergic inflammation and mucosa pathology in these models. Concomitant respiratory syncytial virus infection significantly enhanced allergen sensitization by aerosol exposure and exacerbated eosinophilic inflammation and airway mucosa pathology. Depletion of IgE in this model significantly reduced lung eosinophilic inflammation and airway mucosa pathology. However, in the model in which animals were sensitized by means of systemic allergen exposure with adjuvant, depletion of IgE had no ameliorative effect on lung inflammation or pathology. We demonstrated that the method of antigen sensitization can delineate the role of IgE in allergen-induced lung inflammation. In a murine model that more closely resembles ambient allergen exposure in human subjects, IgE had a critical role in the pathogenesis of allergic asthma and mucosa pathology. The results parallel the results reported with anti-IgE efficacy in allergic asthmatic human subjects.


Silicon analysis of breast and periprosthetic capsular tissue from patients with saline or silicone gel breast implants.
The ubiquitousness of silicon is well known. Recent work has demonstrated measurable baseline levels of silicon in nonaugmented cadavers, subsequent to numerous reports of significant elevations of such levels within patients with silicone breast implants and even more reports alleging a causal relation between silicone gel prostheses and connective-tissue diseases. Despite the lack of scientifically substantiated data that such a relation exists, the calamitous silicone breast implant controversy has ensued. Saline-filled breast implants are constructed with a silicone elastomer envelope that remains in direct contact with periprosthetic capsular tissue following implantation. Although there is no evidence to link saline implants with any disorders, it is important to know if saline breast implants contribute any silicon to human body baseline silicon levels. The present study measured tissue silicon levels in 28 breasts of 16 patients with saline-filled implants to determine if the silicone envelope of these prostheses can contribute to the elevation of such levels. These data were compared with data from 116 breasts of 65 patients with silicone gel-filled prostheses as well as breast tissue from 17 patients (controls) who had never been exposed to either type of implant. Samples of breast tissue and periprosthetic capsular tissue were obtained from patients with both intact and ruptured implants. Silicon levels of breast tissue specimens from patients with saline-filled implants were within the range of the controls if the implants were intact. Silicon levels in periprosthetic capsular tissue from patients with intact saline-filled implants were significantly higher than controls (p < 0.02); however, they were still 100-fold less than capsular tissue levels from patients with intact gel-filled implants. Silicon levels measured in both types of tissue were significantly elevated in patients with silicone gel-filled implants compared with controls (p < 0.01). In the case of ruptured gel implants, breast tissue demonstrated higher silicon levels than did similar specimens from patients with intact implants (p < 0.054); periprosthetic capsular tissue levels also were elevated, although the differences were not statistically significant (p = 0.54). These findings are independent of the implant brand or length of exposure to the particular prosthesis. The finding of elevated levels of silicon in both breast and periprosthetic capsular tissue in patients with silicone gel-filled implants in no way implies or substantiates any claim of a causal relationship between silicone and any reported illnesses.


Analysis of miniature FLIR test targets.
Hughes Aircraft Company has developed a miniature forward-looking infrared test target that uses differential emissivity to simulate differential temperature. This paper provides a theoretical analysis for the emissivity target. The analysis leads to a semiempirical formula that gives a reasonably good prediction of apparent temperature differences. The formula is useful for designing future emissivity targets.


[Metastatic penile tumor from lung cancer: a case report].
A 70-year-old man, who had squamous cell carcinoma of lung, complained of urethral pain. Physical examination revealed an induration of the bulbous urethra and a month later corpora cavernosa were involved. Biopsy was performed and pathological diagnosis was metastatic penile tumor originating from squamous cell carcinoma of lung. Several days after the biopsy the patient died of pleuritis carcinomatosa. Metastatic penile tumor is a rare disease and only 87 cases have been reported in Japan. We review the literature on metastatic penile tumors.


Submicroscopic changes of cortical capillary pericytes in human perifocal brain edema.
Human brain capillary pericytes were studied by electron microscopy. Cortical biopsies from 17 patients with brain edema associated to brain trauma, tumors and congenital malformations were analysed. The data obtained in this study showed that pericytes like endothelial cells exhibited remarkable edematous changes, increased vacuolar and vesicular transport, formation of transient transpericytal channels and tubular structures, demonstrating pericyte brain barrier dysfunction. Images of normal and contracted pericytes showing shallow and deep nuclear invaginations, nuclear basal lamina, actin and myosin-like filaments and coupled interaction with endothelial cells through macula occludens were also encountered. Some pericytes displayed hypertrophic and necrotic changes. Phagocytic pericytes exhibited ingested erythrocytes. Hypertrophic pericytes inducing basement membrane splitting and degenerated pericytes with lacunar enlargement of endoplasmic reticulum, deposit of dense osmiophilic bodies and glycogen granules, vacuolization, hydropic changes of Golgi apparatus, and pleomorphic mitochondria were also found. Certain micropinocytotic vesicles seem to be orientated to multivesicular bodies suggesting that pericytes might play some role in edema resolution. Evidence of pericyte transformation into microglial cells or division into phagocytic cells and pericyte migration were not found.


OP-1 application in bone allograft integration: preliminary results in sheep experimental surgery.
Massive bone allografts are frequently used in orthopaedic reconstructive surgery. However the failure rate at long term follow-up is around 25%. Stimulation of allograft incorporation. In order to stimulate bone remodeling of an allograft we applied recombinant human osteogenic protein-1 (rh-OP-1, also know as bone morphogenetic protein-7, BMP-7) to a long bone critical size defect sheep model. In nine sheep we created a 3 cm osteoperiosteal metatarsal defect replaced with a structural allograft alone (control group, 4 animals), or an allograft added with rh-BMP-7 (BMP group, 5 animals). Radiographic, mechanical, histological and histomorphometric analysis were performed. X-rays in the BMP group showed a better and faster callus formation, compared to the control group within the first 8 weeks after surgery. After 16 weeks there was a higher evidence of bone remodeling in the BMP group. Radiographic healing at junction sites was more evident in the BMP group at 4, 8 and 16 weeks. Mechanical testing on screw extraction showed no statistical differences between the two groups and histomorphometry showed no difference in terms of newly formed bone inside the allograft as well. The resorption rate of the graft was higher in the BMP group in comparison to the control group. The penetration of newly formed vessels was significantly higher in the BMP group. These findings indicate that BMP-7 added to a structural bone allograft inducing early remodeling of the graft through stimulation of neo-angiogenesis and osteoclastic activity, without negative effects in mechanical strength and clinical outcome.


Reverse transcribing the code for chromosome stability.
The linearity of eukaryotic chromosomes presents challenges to cells, as the presence of DNA "ends" poses problems for the DNA replication machinery and the cell's damage response systems. This year's Nobel Prize in Physiology or Medicine recognized groundbreaking studies establishing the telomere field as a crucial area of biomedical research.


A demographic-economic explanation of political stability: Mauritius as a microcosm.
"This paper examines current models of economic and political development--social modernization theory, political and economic characteristics of stable regimes, and cross country analysis of political stability--and tests them on the Indian Ocean Island of Mauritius. The analysis continues with a causal explanation for political stability in Mauritius' recent history, derived from an examination of economic policies and demographic patterns. Political change in Mauritius over the past sixty years seems to be explained best by a model for political stability which integrates specific economic and demographic factors. The model, applicable to development in other third world nations, revises Malthus' conclusion that population and economic conditions move in an oscillatory relationship and replaces it with a more comprehensive theory, suggesting that political stability is a function of both economic development and a repeating cyclical relationship between economics and population."


Neuroprotection by taurine in ethanol-induced apoptosis in the developing cerebellum.
Acute ethanol administration leads to massive apoptotic neurodegeneration in the developing central nervous system. We studied whether taurine is neuroprotective in ethanol-induced apoptosis in the mouse cerebellum during the postnatal period. The mice were divided into three groups: ethanol-treated, ethanol+taurine-treated and controls. Ethanol (20% solution) was administered subcutaneously at a total dose of 5 g/kg (2.5 g/kg at time 1 h and 2.5 g/kg at 3 h) to the ethanol and ethanol+taurine groups. The ethanol+taurine group also received two injections of taurine (1 g/kg each, at time zero and at 4 h). To estimate apoptosis, immunostaining for activated caspase-3 and TUNEL staining were made in the mid-sagittal sections containing lobules I-X of the cerebellar vermis at 12 or 8 hours after the first taurine injection. Changes in the blood taurine level were monitored at each hour by reverse-phase high-performance liquid chromatography (HPLC). Ethanol administration induced apoptosis of Purkinje cells on P4 in all cerebellar lobules, most extensively in lobules IX and X, and on P7 increased the number of activated caspase-3-immunoreactive and TUNEL-positive cells in the internal layer of the cerebellum. Administration of taurine significantly decreased the number of activated caspase-3-immunoreactive and TUNEL-positive cells in the internal layer of the cerebellum on P7, but had no effect on Purkinje cells in P4 mice. The high initial taurine concentration in blood of the ethanol+taurine group diminished dramatically during the experiment, not being different at 13 h from that in the controls. We conclude that the neuroprotective action of taurine is not straightforward and seems to be different in different types of neurons and/or requires prolonged maintenance of the high taurine concentration in blood plasma.


Antibodies to beta 1 and beta 2 adrenoreceptors in Chagas' disease.
Evidence accumulated over the last decade concerning human and experimental models suggests that an immunopathological mechanism may be involved in the pathogenesis of chronic Chagas' disease. In this paper we demonstrate the existence of two different circulating IgG in chagasic patients which bind with myocardial beta 1 and spleen cell beta 2 adrenoceptors, acting as non-competitive inhibitors. Both chagasic IgG against beta 1 and beta 2 adrenoceptor increased intracellular levels of cAMP that could be blocked by specific beta 1 and beta 2 adrenoceptor antagonists. The specificity for beta 1 and beta 2 adrenoceptors and the independence of other tissue reactive antibodies was demonstrated by IgG absorption with turkey red blood cell (TRBC), human lymphocytes (HL) or guinea pig red blood cells (GPRBC). The F(ab')2 fraction acted similarly. This supports the specificity of beta 1 and beta 2 adrenoceptors to the chagasic IgG and the independence of the other tissue reactive antibodies, such as EVI system. The probable pathogenic role of both beta 1 and beta 2 adrenergic chagasic antibody is discussed.


Observation of Cytoplasmic and Vacuolar Malate in Maize Root Tips by C-NMR Spectroscopy.
The accumulation of malate by maize (Zea mays L.) root tips perfused with KH(13)CO(3) was followed by (13)C nuclear magnetic resonance spectroscopy. In vivo nuclear magnetic resonance spectra contained distinct signals from two pools of malate in maize root tips, one at a pH approximately 5.3 (assigned to the vacuole) and one at a pH > 6.5 (assigned to the cytoplasm). The ratio of cytoplasmic to vacuolar malate was lower in 12 millimeter long root tips than in 2 millimeter root tips. The relatively broad width of the signals from C1- and C4-labeled vacuolar malate indicated heterogeneity in vacuolar pH. During the 3 hour KH(13)CO(3) treatment, (13)C-malate accumulated first primarily in the cytoplasm, increasing to a fairly constant level of approximately 6 millimolar by 1 hour. After a lag, vacuolar malate increased throughout the experiment.


Prevalence of Overactive Bladder Symptoms Among Women With Interstitial Cystitis/Bladder Pain Syndrome.
Symptoms of urinary frequency, urgency, and urinary leakage are characteristic of overactive bladder (OAB) syndrome. However, frequency and urgency symptoms are also present in most patients with interstitial cystitis/bladder pain syndrome (IC/BPS). Our objective was to describe the urge incontinence among women with IC/BPS, which may indicate true overlap of OAB and IC/BPS. This is a prospective study of women with IC/BPS diagnosed clinically in the Veterans Affairs Health Care system. Patients completed the OAB and Female Genitourinary Pain Index (F-GUPI) questionnaires. Questions from the OAB questionnaire were used to analyze symptoms of urinary urgency and urge incontinence. Pain symptoms, urinary symptoms, and impact on quality of life were assessed based on the F-GUPI. Patient demographics, comorbidities, and symptoms were reviewed. Within the cohort of 144 women with IC/BPS, 100 (69%) had urinary leakage associated with the strong desire to void and were more likely to have incontinence compared with healthy controls (P < 0.001). The IC/BPS group also had higher total and pain scores on the F-GUPI (P < 0.001), but pain scores were not affected by the presence of incontinence (P = 0.478). The prevalence of OAB symptoms of urinary leakage is high among women with IC/BPS. This may explain the efficacy of OAB medication and third-line therapies in this population.


Selection and Warning of Evidence-Based Antidiabetic Medications for Patients With Chronic Liver Disease.
The global prevalence of chronic liver disease and diabetes mellitus (DM) has gradually increased potentially due to changes in diet and lifestyle. The choice of antidiabetic medications for patients with coexisting DM and chronic liver disease is complicated. Severe liver injury may decrease the metabolism of antidiabetic medications, resulting in elevated drug concentrations and adverse effects. The choice of antidiabetic medications in patients with chronic liver disease has not been well studied. The long-term outcomes of antidiabetic medications in patients with chronic liver disease have gained attention recently. Herein, we reviewed relevant articles to extend our understanding on the selection and warning of antidiabetic medications for patients with chronic liver disease.


Potential for abuse in the VCT counselling room: service provider's perceptions in Kenya.
The rapid scale-up of HIV counselling and testing programmes in Kenya has led to quality concerns, including the potential for abuse within the private, confidential setting of client-initiated voluntary counselling and testing (VCT). A qualitative study was conducted in three provinces of Kenya, involving 26 VCT service providers and 13 key informants. First and second hand accounts of emotional, physical and sexual abuse emerged in all three study sites in spite of measures to mitigate such occurrences. Whilst uncommon, abuse was perceived by service providers to be serious and sufficiently widespread to raise significant concerns. Abuse occurred client to counsellor, from counsellor to client and from counsellor to counsellor. In all cases the person suffering the abuse was female. While the potential for abuse was demonstrated in VCT sites, we argue that experiences of abuse are not confined to VCT and are largely shaped by gender and power relations within the Kenyan cultural context. The international impetus for scale-up of HIV services provides an urgent rationale for the need to address and highlight these difficult issues at multiple levels. International guidelines, policy and methods need adapting in recognition of the potential for abuse. Systems for investigating and deregistering counsellors have been developed in Kenya but require formalizing. Institutions providing VCT should consider unlocked doors, semi-opaque windows and the use of 'mystery clients' as a quality assurance measure.


A preparative method for crosslinking proteins to DNA in nuclei by single-pulse UV laser irradiation.
Photocrosslinking of proteins to DNA by single-pulse UV laser has been used only in analytical experiments, carried out with reconstituted complexes of a single DNA binding protein and a labeled target sequence. Here we propose a large-scale technique for irradiation of nuclei, generating preparative quantities of covalently linked protein-DNA complexes for further analysis of the partner molecules. The use of a flow cuvette allows a milligram of DNA in either nuclei or chromatin to be irradiated by a single pulse within few minutes. The efficiency of crosslinking varies from 6 to 12% of the total nuclear proteins. The presence of histones and other chromosomal proteins in the crosslinked protein-DNA complexes was demonstrated by using specific antibodies. The irradiation procedure can be fully automated using a microcomputer.


In vitro measurement accuracy of an electronic apex locator in teeth with simulated apical root resorption.
The purpose of this in vitro study was to evaluate the accuracy of Root ZX apex locator to determine the working length in teeth with simulated apical root resorption. Fifty extracted, single-rooted, human teeth with mature apices were used in this study. An irregular cavity defect was drilled at the apex of each tooth simulating an apical root resorption. Three operators used the Root ZX to measure the working lengths, comparing the electronic readings with the direct visual measurements. The Root ZX was 62.7%, 94.0%, and 100.0% accurate to within 0.5 mm, 1 mm, and 1.5 mm of the direct visual measurements, respectively. Statistically significant differences were observed between operator A and B and A and C (p < 0.01), but no significant differences were detected between operator B and C (p > 0.01).


Statistical inference by confidence intervals: issues of interpretation and utilization.
This article examines the role of the confidence interval (CI) in statistical inference and its advantages over conventional hypothesis testing, particularly when data are applied in the context of clinical practice. A CI provides a range of population values with which a sample statistic is consistent at a given level of confidence (usually 95%). Conventional hypothesis testing serves to either reject or retain a null hypothesis. A CI, while also functioning as a hypothesis test, provides additional information on the variability of an observed sample statistic (ie, its precision) and on its probable relationship to the value of this statistic in the population from which the sample was drawn (ie, its accuracy). Thus, the CI focuses attention on the magnitude and the probability of a treatment or other effect. It thereby assists in determining the clinical usefulness and importance of, as well as the statistical significance of, findings. The CI is appropriate for both parametric and nonparametric analyses and for both individual studies and aggregated data in meta-analyses. It is recommended that, when inferential statistical analysis is performed, CIs should accompany point estimates and conventional hypothesis tests wherever possible.


Effects of CYP3A4 inhibition and induction on the pharmacokinetics and pharmacodynamics of tolvaptan, a non-peptide AVP antagonist in healthy subjects.
Before these trials were done, the effects of CYP3A4 inhibition and induction on the pharmacokinetics (PK) and pharmacodynamics (PD) of tolvaptan in healthy subjects were unknown. As tolvaptan is a CYP3A4 substrate, knowing the effects of inhibition and induction on CYP3A4-mediated metabolism was important for dosing recommendations. This paper describes the changes in tolvaptan PK and PD following inhibition or induction of CYP3A4 and explores the mechanisms behind the disparity seen between tolvaptan PK and effects on urine output. It also discusses the concentrations at which tolvaptan produces its maximal response on urine output and the timing of the onset and offset of this response. AIMS In vitro studies indicated CYP3A4 alone was responsible for tolvaptan metabolism. To determine the effect of a CYP3A4 inhibitor (ketoconazole) and a CYP3A4 inducer (rifampicin) on tolvaptan pharmacokinetics (PK) and pharmacodynamics (PD), two clinical trials were performed. For CYP3A4 inhibition, a double-blind, randomized (5:1), placebo-controlled trial was conducted in 24 healthy subjects given either a single 30 mg dose of tolvaptan (n= 19) or matching placebo (n= 5) on day 1 with a 72 h washout followed by a 3 day regimen of 200 mg ketoconazole, once daily with 30 mg tolvaptan or placebo also given on day 5. For CYP3A4 induction, 14 healthy subjects were given a single dose of 240 mg tolvaptan with 48 h washout followed by a 7 day regimen of 600 mg rifampicin, once daily, with 240 mg tolvaptan also given on the seventh day. When co-administered with ketoconazole, mean C(max) and AUC(0,∞) of tolvaptan were increased 3.48- and 5.40-fold, respectively. Twenty-four hour urine volume increased from 5.9 to 7.7 l. Erythromycin breath testing showed no difference following a single dose of tolvaptan. With rifampicin, tolvaptan mean C(max) and AUC were reduced to 0.13- and 0.17-fold of tolvaptan administered alone. Twenty-four hour urine volume decreased from 12.3 to 8.8 l. Tolvaptan is a sensitive CYP3A4 substrate with no inhibitory activity. Due to the saturable nature of tolvaptan's effect on urine excretion rate, changes in the pharmacokinetic profile of tolvaptan do not produce proportional changes in urine output.


Urinary soluble HLA-DR is a potential biomarker for acute renal transplant rejection.
BACKGROUND.: Urine is a potentially rich source of biomarkers for monitoring kidney dysfunction. In this study, we have investigated the potential of soluble human leukocyte antigen (sHLA)-DR in the urine for noninvasive monitoring of renal transplant patients. METHODS.: Urinary soluble HLA-DR levels were measured by sandwich enzyme-linked immunosorbent assay in 103 patients with renal diseases or after renal transplantation. sHLA-DR in urine was characterized by Western blotting and mass spectrometry. RESULTS.: Acute graft rejection was associated with a significantly elevated level of urinary sHLA-DR (P<0.0001), compared with recipients with stable graft function or healthy individuals. A receiver operating characteristic curve analysis showed the area under the curve to be 0.88 (P<0.001). At a selected threshold, the sensitivity was 80% and specificity was 98% for detection of acute renal transplant rejection. sHLA-DR was not exosomally associated and was of lower molecular weight compared with the HLA-DR expressed as heterodimer on the plasma membrane of antigen-presenting cells. CONCLUSIONS.: sHLA-DR excreted into urine is a promising indicator of renal transplant rejection.


Vascular and fibrinolytic effects of intra-arterial tumour necrosis factor-alpha in patients with coronary heart disease.
Elevated plasma t-PA (tissue plasminogen activator) and serum CRP (C-reactive protein) concentrations are associated with an adverse cardiovascular risk. In the present study, we investigated whether acute local inflammation causes vascular dysfunction and influences t-PA release in patients with stable coronary heart disease. Serum CRP, plasma t-PA and PAI-1 (plasminogen activator inhibitor type 1) concentrations were determined in 95 patients with stable coronary heart disease. A representative subpopulation of 12 male patients received an intra-brachial infusion of TNF-alpha (tumour necrosis factor-alpha) and saline placebo using a randomized double-blind cross-over study design. Forearm blood flow and plasma fibrinolytic and inflammatory variables were measured. Serum CRP concentrations correlated with plasma t-PA concentrations (r=0.37, P<0.001) and t-PA/PAI-1 ratio (r=-0.21, P<0.05). Intra-arterial TNF-alpha caused a rise in t-PA concentrations (P<0.001) without affecting blood flow or PAI-1 concentrations. TNF-alpha pretreatment impaired acetylcholine- and sodium nitroprusside-induced vasodilatation (P<0.001 for both) whilst doubling bradykinin-induced t-PA release (P=0.006). In patients with stable coronary heart disease, plasma fibrinolytic factors correlate with a systemic inflammatory marker and local vascular inflammation directly impairs vasomotor function whilst enhancing endothelial t-PA release. We suggest that the adverse prognosis associated with elevated plasma t-PA concentrations relates to the underlying causative association with vascular inflammation and injury.


The effects of coating culture dishes with collagen on fibroblast cell shape and swirling pattern formation.
Motile human-skin fibroblasts form macroscopic swirling patterns when grown to confluence on a culture dish. In this paper, we investigate the effect of coating the culture-dish surface with collagen on the resulting pattern, using human-skin fibroblast NB1RGB cells as the model system. The presence of the collagen coating is expected to enhance the adherence of the fibroblasts to the dish surface, and thereby also enhance the traction that the fibroblasts have as they move. We find that, contrary to our initial expectation, the coating does not significantly affect the motility of the fibroblasts. Their eventual number density at confluence is also unaffected. However, the coherence length of cell orientation in the swirling pattern is diminished. We also find that the fibroblasts cultured in collagen-coated dishes are rounder in shape and shorter in perimeter, compared with those cultured in uncoated polystyrene or glass culture dishes. We hypothesise that the rounder cell-shape which weakens the cell-cell nematic contact interaction is responsible for the change in coherence length. A simple mathematical model of the migrating fibroblasts is constructed, which demonstrates that constant motility with weaker nematic interaction strength does indeed lead to the shortening of the coherence length.


Double-layered free temporal fascia flap as a two-layered tendon-gliding surface.
We report the use of a two-layered free fascial flap consisting of temporoparietal and deep temporal fascia based on a single vascular pedicle, the superficial temporal artery and vein. The flap was used to reconstruct an extensive degloving injury of the dorsum of the hand, in which multiple intact extensor tendons lay fully exposed on all sides, with exposed bone beneath them. By sandwiching the tendons between the layers of vascularized fascia, gliding surfaces were provided, both superficial and deep to the exposed tendons. The single-stage reconstruction was completed with a split-thickness skin graft. The patient returned to heavy manual work within 12 weeks of injury. He obtained an excellent range of movement without the need for tenolysis.


Expression and function of Wnt5a in the development of the glandular stomach in the chicken embryo.
The epithelium of the chicken embryonic glandular stomach (proventriculus) differentiates into both a glandular and a luminal epithelium, the cells of which express specific marker genes. The subsequent formation and differentiation of the glands then proceed under the influence of the mesenchyme. To search for possible candidates for the mesenchymal factors involved, we have now investigated the expression and function of Wnt5a in this process. Our current results show that Wnt5a is expressed in the mesenchyme during active gland formation and that overexpression of this gene in ovo results in the increased and ectopic expression of some of the marker genes of the luminal and glandular epithelia. In particular, the overexpression of Wnt5a markedly enhances the expression of the embryonic chicken pepsinogen gene, a marker of the glandular epithelium, indicating its role as a mesenchymal factor that regulates the differentiation of the proventricular epithelium.


Histamine in two component system-mediated bacterial signaling.
Histamine is a key mediator governing vital cellular processes in mammals beyond its decisive role in inflammation. Recent evidence implies additional actions in both eukaryotes and prokaryotes. Besides its function in host defense against bacterial infections, histamine elicits largely undefined actions in microorganisms that may contribute to bacteria-host interactions. Bacterial proliferation and adaptation are governed by sophisticated signal transduction networks, including the versatile two-component systems (TCSs) that comprise sensor histidine kinases and response regulators and rely on phosphotransfer mechanisms to exert their modulatory function. The AtoSC TCS regulates fundamental cellular processes such as short-chain fatty acid metabolism, poly-(R)-3-hydroxybutyrate (cPHB) biosynthesis and chemotaxis in Escherichia coli. The implication of exogenous histamine in the AtoSC-mediated cPHB biosynthesis and in E. coli chemotactic behavior is indicative of a putative function of histamine in bacterial physiology. The data raise questions on the significance of histamine actions in bacteria-host symbiosis, dysbiosis and pathogenicity as well as on the possible consequences upon therapeutic administration of histamine receptor-targeting agents and in particular ligands of the recently identified immunomodulatory H4 receptor.


Identification of distinct lymphocyte subsets responding to subcellular fractions of Mycobacterium bovis bacille calmette-Guérin (BCG).
In order to investigate the ability of Mycobacterium bovis BCG vaccination to induce immune responses toward different classes of mycobacterial antigens and the cell populations involved in such responses, proliferation of distinct human lymphocyte subsets from BCG-vaccinated donors in response to different subcellular fractions of BCG was analysed and compared with that of not sensitized subjects. Proliferation of different cell subsets was evaluated by flow cytometric determination of bromodeoxyuridine incorporated into DNA of dividing cells and simultaneous identification of cell surface markers. Although a certain degree of variability was observed among different donors, after 6 days of in vitro stimulation BCG-vaccinated subjects displayed, as a mean, a stronger blastogenic response to all the classes of antigens compared with non-sensitized ones. PPD, culture filtrates and membrane antigens induced a predominant proliferation of CD4+ T cells. In contrast, preparations enriched in cytosolic antigens elicited strong proliferation of gammadelta+ T cells which, as a mean, represented 55% of the proliferating cells. Although to a lesser extent, proliferation of gammadelta+ T cells was also elicited by preparations enriched in membrane and cell wall antigens. In response to the latter preparation proliferation of CD4+ T cells and CD16+/CD3- (natural killer (NK)) cells was observed, as well. In particular, cell wall antigens were found to induce significantly higher levels of proliferation of NK cells compared with all the other classes of antigens.


Edge enhancement and image equalization by unsharp masking using self-adaptive photochromic filters.
A new method for real-time edge enhancement and image equalization using photochromic filters is presented. The reversible self-adaptive capacity of photochromic materials is used for creating an unsharp mask of the original image. This unsharp mask produces a kind of self filtering of the original image. Unlike the usual Fourier (coherent) image processing, the technique we propose can also be used with incoherent illumination. Validation experiments with Bacteriorhodopsin and photochromic glass are presented.


Apparently normal kidney development in mice with conditional disruption of ANG II-AT1 receptor genes in FoxD1-positive stroma cell precursors.
An intact renin-angiotensin system involving ANG II type 1 (AT1) receptors is crucial for normal kidney development. It is still unclear in which cell types AT1 receptor signaling is required for normal kidney development, maturation, and function. Because all kidney cells deriving from stroma progenitor cells express AT1 receptors and because stromal cells fundamentally influence nephrogenesis and tubular maturation, we investigated the relevance of AT1 receptors in stromal progenitors and their descendants for renal development and function. For this aim, we generated and analyzed mice with conditional deletion of AT1A receptor in the FoxD1 cell lineage in combination with global disruption of the AT1B receptor gene. These FoxD1-AT1ko mice developed normally. Their kidneys showed neither structural nor functional abnormalities compared with wild-type mice, whereas in isolated perfused FoxD1-AT1ko kidneys, the vasoconstrictor and renin inhibitory effects of ANG II were absent. In vivo, however, plasma renin concentration and renal renin expression were normal in FoxD1-AT1ko mice, as were blood pressure and glomerular filtration rate. These findings suggest that a strong reduction of AT1 receptors in renal stromal progenitors and their descendants does not disturb normal kidney development.


Bacillus-Mediated Silver Nanoparticle Synthesis and Its Antagonistic Activity against Bacterial and Fungal Pathogens.
In this article, the supernatant of the soil-borne pathogen Bacillus mn14 was used as the catalyst for the synthesis of AgNPs. The antibacterial and antifungal activity of Bs-AgNPs was evaluated, in which S. viridans and R. solani showed susceptibility at 70 µL and 100 µL concentrations. Enzyme properties of the isolates, according to minimal inhibitory action and a growth-enhancing hormone-indole acetic acid (IAA) study of the isolates, were expressed in TLC as a purple color with an Rf value of 0.7. UV/Vis spectroscopy revealed the presence of small-sized AgNPs, with a surface plasmon resonance (SPR) peak at 450 nm. The particle size analyzer identified the average diameter of the particles as 40.2 nm. The X-ray diffraction study confirmed the crystalline nature and face-centered cubic type of the silver nanoparticle. Scanning electron microscopy characterized the globular, small, round shape of the silver nanoparticle. AFM revealed the two-dimensional topology of the silver nanoparticle with a characteristic size ranging around 50 nm. Confocal microscopy showed the cell-wall disruption of S. viridans treated with Bs-AgNPs. High-content screening and compound microscopy revealed the destruction of mycelia of R. solani after exposure to Bs-AgNPs. Furthermore, the Bs-AgNPs cured sheath blight disease by reducing lesion length and enhancing root and shoot length in Oryza sativa seeds. This soil-borne pathogen Bacillus-mediated synthesis approach of AgNPs appears to be cost-efficient, ecofriendly, and farmer-friendly, representing an easy way of providing valuable nutritious edibles in the future.


Leptin and leptin receptor expression in rat and mouse pituitary cells.
Leptin is a circulating hormone secreted mainly by adipose tissue. Recent studies have shown leptin production by other tissues, including the placenta, stomach, and mammary tissues. Various reports have suggested that the anterior pituitary may have a role in the regulatory effects of leptin. We recently localized leptin in the human anterior pituitary, but analysis of leptin in rodent pituitary has not been previously reported. In this study we examined rat and mouse pituitary tissues and various cell lines for leptin by RT-PCR, immunohistochemistry, and Western blotting. Leptin receptor messenger RNA was also examined in these tissues by RT-PCR. Leptin was present in a small percentage of rat (4.8 +/- 0.7%) and mouse (7 +/- 2%) pituitary cells. Colocalization studies with leptin and pituitary hormones showed leptin expression mainly in TSH cells (24 +/- 2% of TSH cells in the rat pituitary and 31 +/- 1% of TSH cells in the mouse pituitary). A folliculo-stellate (FS) cell line, TtT/GF, also expressed leptin. The long isoform of leptin receptor (OB-Rb) was present in normal pituitary and in various pituitary cell lines, including FS, GH3, and alphaT3-1 cells. Treatment of GH3 and FS cells with leptin (1 x 10(-8) M) inhibited cell proliferation assessed by [3H]thymidine incorporation in GH3, but not in FS, cells. These findings show for the first time that leptin is expressed in rat and mouse anterior pituitaries mainly by TSH cells and by a mouse FS cell line. The finding of leptin and of the long isoform of leptin receptor in normal rat and mouse pituitaries and in various cell lines implicates an autocrine/paracrine loop in the production and regulation of leptin and leptin receptor in the rodent pituitary.


Neuroinvasive potential of SARS-CoV2 with neuroradiological and neuropathological findings: is the brain a target or a victim?
Coronaviridae family includes pathogen viruses for humans, that lead to clinical conditions with main respiratory involvement; many of these viruses have notoriously a neuroinvasive potential, as demonstrated by published data on SARS-CoV and MERS-CoV epidemics, as well by results obtained in experimental models. During pandemic of coronavirus disease 2019 (COVID-19), it is noticed that the central nervous system involvement represented a truly significant moment in the history of some COVID-19 patients; indeed, clinical and radiological features published in literature regarding COVID-19 disease are consistent with a neurological involvement. It is also known that histopathological data related to SARS-CoV2 infection have been published with considerable delay, which was even greater for neuropathological information. Moreover, many published data are incomplete, and often the lesions described are not directly related to the action of the virus. In this review, we collected the available radiological and neuropathological information, in order to delineate a more complete picture of the relationship between SARS-Cov2 and brain, focusing our attention on the two most important neuroinvasion routes for the virus. We also highlighted what we consider methodological mistakes both in the autopsy procedures and brain study in COVID-19 deaths. We emphasize the need for a complete study of all the organs in case of autopsy. It is important that through this experience, we no longer do the mistake of neglecting the brain.


A tree of tree frogs around the Black Sea.
Speciation, the process by which one species evolves into two or more, is a major focus of ongoing debate, particularly regarding the geographic context in which it occurs. Geographic models of speciation tend to fall into discrete categories, typically referred to as allopatric, parapatric and sympatric speciation, according to whether two groups evolve reproductive isolation while geographically isolated, differentiated but connected by gene flow, or completely co-occurring. Yet molecular studies indicate that full development of reproductive isolation can take very long compared with the timescale at which climatic oscillations occur, such that the geographic context of differentiating forms might change often during the long process to full species. Studies of genetic relationships across the ranges of organisms with low-dispersal distances have the potential to reveal these complex histories. In a particularly elegant example in this issue, Dufresnes et al. () use genetic variation and ecological niche modelling to show that a ring of populations of the eastern tree frog (Hyla orientalis) surrounding the Black Sea had a complex history of geographic differentiation. Alternating phases of geographic fragmentation and phases of gene flow between neighbouring populations have produced a pattern of gradual genetic change connecting the western, southern and eastern sides of the ring, with the northwestern and northeastern forms being most differentiated. In the north, a population in Crimea appears to have been produced through mixture of the two extreme forms. The overall genetic relationships are reminiscent of those found in ring species, which have been used as prime demonstrations of the process of speciation. The difference, however, is that the terminal forms appear to have mixed rather than be reproductively isolated, although more research is needed to infer whether there might be some reproductive isolation on the northern side of the ring.


External quality assessment of sputum smear microscopy in Taiwan.
To evaluate the quality of sputum smear microscopy in nine Taiwan Centers for Disease Control contract laboratories, an external quality assessment (EQA) programme has been implemented since 2005. A sampling strategy based on the lot quality assurance system was applied to select slides for rechecking. Supervisory visits and technical training were conducted to determine the causes of errors and to take corrective action. Of the 1017 slides sampled in 2005, 637 (63%) had proper smear size, 492 (48%) proper thickness and 884 (87%) proper staining; the corresponding figures were 972 (100%), 748 (77%) and 809 (99.6%) for the 972 slides rechecked in 2006. After training, the quality of size and staining of smear preparation had significantly improved (P < 0.001) in 2006. Rechecking of 981 readable slides in 2005 identified 3 (0.3%) high false-negatives, 3 (16.7%) low false-positives and 26 (2.8%) low false-negatives; the corresponding errors were 3 (0.3%), 8 (28.6%) and 12 (1.3%) for the 972 slides rechecked in 2006. Of the eight laboratories, two (25%) and four (50%) reached 80% sensitivity in 2005 and 2006, respectively. Technical training and EQA improved the quality of sputum smear microscopy services.


Antibacterial Activity of T22, a Specific Peptidic Ligand of the Tumoral Marker CXCR4.
CXCR4 is a cytokine receptor used by HIV during cell attachment and infection. Overexpressed in the cancer stem cells of more than 20 human neoplasias, CXCR4 is a convenient antitumoral drug target. T22 is a polyphemusin-derived peptide and an effective CXCR4 ligand. Its highly selective CXCR4 binding can be exploited as an agent for the cell-targeted delivery and internalization of associated antitumor drugs. Sharing chemical and structural traits with antimicrobial peptides (AMPs), the capability of T22 as an antibacterial agent remains unexplored. Here, we have detected T22-associated antimicrobial activity and biofilm formation inhibition over Escherichia coli, Staphylococcus aureus and Pseudomonas aeruginosa, in a spectrum broader than the reference AMP GWH1. In contrast to GWH1, T22 shows neither cytotoxicity over mammalian cells nor hemolytic activity and is active when displayed on protein-only nanoparticles through genetic fusion. Under the pushing need for novel antimicrobial agents, the discovery of T22 as an AMP is particularly appealing, not only as its mere addition to the expanding catalogue of antibacterial drugs. The recognized clinical uses of T22 might allow its combined and multivalent application in complex clinical conditions, such as colorectal cancer, that might benefit from the synchronous destruction of cancer stem cells and local bacterial biofilms.


[Contribution of 19th-century religious congregations in the development of the nursing profession].
The purpose of this historical research project is to examine the contribution of 19th-century religious congregations in the development of the nursing profession, based on the historical example of the Sisters of Charity of Sts. Bartolomea Capitanio and Vincenza Gerosa (or the Sisters of the Holy Child Mary). To this end, sixty three volumes were analysed, all taken from the historical archive of the Generalate of the Sisters of the Holy Child Mary in Milan, in via S. Sofia n.13, with the exception of just one, taken from the Braidense National Library, also in Milan. This research project has highlighted the sociological contribution provided by the Sisters of the Holy Child Mary to the professional nature of nursing, and to the development of the distinctive features of the nursing profession (Greenwood, 1980). All documentary sources were analysed in line with the Chabod historical research method (2006), and for their critical interpretation, a scheme of analysis was created. Two lines of investigation emerged from the data collected: the role of Sister Emilia Vinante as an expert with regard to the nursing profession, and the professional strategies promoted by FIRO (Federation of Italian Religious Nurses). Based on the conclusions of the research project, it may be stated that religious congregations contributed greatly to the nursing profession, leaving a decisive mark on the cultural and professional development of nurses.


Experimental study of two methods of data collection by questionnaire.
The aim of the present study was to compare the results obtained using two different methods of data collection about caries preventive services provided in general dental practice. A questionnaire was mailed to a random national sample of 479 dentists resident in Norway in January 1985. The sample was divided into two groups by random allocation. All dentists, irrespective of group, were requested to give background information. One group, comprising 287 dentists (GR), was asked to complete a separate form for every adult patient (greater than or equal to 20 yr) treated in the course of 1 day. The demographic characteristics and dental visiting habits of the patients, as well as the number of teeth present, caries lesions and preventive services rendered were recorded. The other group, 192 dentists (GE), was requested to make general estimates of the time spent on caries prevention and the proportion of patients receiving various types of caries preventive services. The dentists were unaware of the methodologic aspect of the survey and everyone received one reminder in order to guarantee anonymity. The estimation method (GE) did not give the expected advantage over the registration method (GR) in response rate (51.7% vs 46.2%, P greater than 0.40), and gave a gross overestimation of the frequency with which adult patients received different types of caries preventive procedures (P less than 0.005). Thus, even though the estimates of the proportion of total treatment time spent on caries prevention were comparable for the two methods, and the estimation approach is labor-saving, it cannot be recommended for the collection of data on caries prevention in the dental office.


Maximal electrogram attenuation recorded from mini electrodes embedded on 4.5-mm irrigated and 8-mm nonirrigated catheters signifies lesion maturation.
The electrograms (EGMs) recorded from mini electrodes (ME) placed on the tip of the ablation electrode allow more precise EGM monitoring during lesion formation. Our objective was to define the lesion boundaries and extracardiac injuries resulting from 60-second RF application versus RF application time titrated to maximal attenuation of the ME EGM in the atria and ventricles using 4.5-mm irrigated and 8-mm catheters. RF lesions were placed in both atria and ventricles in 13 (30-35 kg) canines; 6 (4.5-mm OI) and 7 (8 mm). The RF application time was fixed at 60 seconds or terminated at maximal ME EGM amplitude attenuation. Pre/postablation pacing thresholds, EGM amplitudes, and lesion dimensions were not significantly different between maximal EGM attenuation and 60-second RF application using either catheter. Atrial lesion transmurality was also similar for both catheters and groups 91.2% (4.5 mm) and 96% (8 mm) when the RF was titrated to the maximal EGM attenuation and 94.2% (4.5 mm) and 95% (8 mm) with 60-second RF. The 60-second RF ablation, however, presented with significant extracardiac injuries to the lungs and esophagus, along with char formation. Deep ventricular lesions were noted with maximal EGM attenuation that were not different from the 60-second RF ablation. Titration of the RF application time to the maximal EGM attenuation based on the ME recordings represents atrial lesion maturation and deep ventricular lesions. Prolonging the RF application results in greater extracardiac injury and char formation without increasing lesion size.


Respiratory syncytial virus load, viral dynamics, and disease severity in previously healthy naturally infected children.
Respiratory syncytial virus (RSV) disease severity was thought to be a result of host immunopathology but alternatively may be driven by high-level viral replication. The relationships between RSV load, viral clearance dynamics, and disease severity have not been carefully evaluated. Previously healthy RSV-infected children <2 years old were recruited. RSV load was measured in respiratory secretions by fresh quantitative culture over 3 hospital days. Measures of disease severity were hospital admission, duration of hospitalization, requirement for intensive care, and respiratory failure. Multivariate logistic regression models revealed independent predictors of increased duration of hospitalization: male sex, lower weight, and higher viral load on any day. Viral loads at day 3 were more significantly associated with requirement for intensive care and respiratory failure than were viral loads at earlier time points. Faster RSV clearance was independently associated with shorter hospitalization. These observations challenge the immunopathology-based pathogenesis paradigm. They also have major therapeutic implications, suggesting that application of antiviral agents early in the disease course, even at a time when viral replication is at its highest, might improve subsequent morbidity by significantly lowering viral load and direct viral cytopathic effects, and aborting the potential downstream immunopathology.


Spontaneous motor entrainment to music in multiple vocal mimicking species.
The human capacity for music consists of certain core phenomena, including the tendency to entrain, or align movement, to an external auditory pulse [1-3]. This ability, fundamental both for music production and for coordinated dance, has been repeatedly highlighted as uniquely human [4-11]. However, it has recently been hypothesized that entrainment evolved as a by-product of vocal mimicry, generating the strong prediction that only vocal mimicking animals may be able to entrain [12, 13]. Here we provide comparative data demonstrating the existence of two proficient vocal mimicking nonhuman animals (parrots) that entrain to music, spontaneously producing synchronized movements resembling human dance. We also provide an extensive comparative data set from a global video database systematically analyzed for evidence of entrainment in hundreds of species both capable and incapable of vocal mimicry. Despite the higher representation of vocal nonmimics in the database and comparable exposure of mimics and nonmimics to humans and music, only vocal mimics showed evidence of entrainment. We conclude that entrainment is not unique to humans and that the distribution of entrainment across species supports the hypothesis that entrainment evolved as a by-product of selection for vocal mimicry.


Pigmentation and Soluble Peroxidase Isozyme Patterns of Leaves of Pedilanthus tithymaloides L. variegatus as a Result of Daily Temperature Differences.
In a constant environment with a narrow (less than 8 degrees C) daily temperature difference (deltat), leaves of Pedilanthus tithymaloides L. variegatus usually appeared green. After at least two days of deltat > 10 degrees C, new leaves had become green-white, and a red pigmentation appeared and increased if high deltat conditions were maintained. If plants were returned to a narrow deltat, new green leaves reappeared. Electrophoretic patterns of soluble peroxidase isozymes changed during the color changes. Three groups of electrophoretic bands occurred, and each was related to a characteristic tissue. The development of red color was correlated with the appearance of the group with the fastest electrophoretic mobility and the highest peroxidase activity.


[Characteristics of the restriction profile of chromosomal DNA in strains of Acidithiobacillus ferroxidans, adapted to various oxidation substrates].
Restriction profiles of chromosomal DNA were studied in different Acidithiobacillus ferrooxidans strains grown on medium with Fe2+ and further adapted to another oxidation substrate (S0, FeS2, or sulfide ore concentrates). The restriction endonuclease XbaI digested the chromosomal DNA from different strains into different numbers of fragments of various sizes. Adaptation of two strains (TFBk and TFN-d) to new oxidation substrates resulted in structural changes in XbaI-restriction patterns of their chromosomal DNA. Such changes in the DNA restriction patterns occurred in strain TFBk after the adaptation to precyanidated gravitational pyrite-arsenopyrite concentrate (no. 1) from the Nezhdaninskoe deposit or to copper-containing ore from the Udokanskoe deposit and also in strain TFN-d adapted to untreated pyrite-arsenopyrite concentrate (no. 2) from the Nezhdaninskoe deposit. No changes in the number or size of the XbaI-restriction patterns of chromosomal DNA were revealed in either strain TFBk cultivated on media with pyrite from the Angren and Tulun deposits or in strains TFN-d and TFO grown on media with S0 and pyrite. Neither were changes observed in the XbaI-restriction patterns of the DNA from strain TFV-1, isolated from the copper ore of the Volkovskoe deposit, when Fe2+ was substituted with alternative substrates--S0, pyrite or concentrate no. 2 from the ore of Nezhdaninskoe deposit. In strain TFO, no differences in the XbaI-restriction patterns of the chromosomal DNA were revealed between the culture grown on medium containing concentrate no. 2 or the concentrate of surface-lying ore from Olimpiadinskoe deposit and the culture grown on medium with Fe2+. When strain TFO was cultivated on the ore concentrate from deeper horizons of the Olimpiadinskoe deposit, which are characterized by lower oxidation degree and high antimony content, mutant TFO-2 differing from the parent strain in the chromosomal DNA structure was isolated. The correlation between the lability of chromosomal DNA structure in A. ferrooxidans strains and the physical and chemical peculiarities of the isolation substrate and habitat is discussed.


n-3 polyunsaturated fatty acids: update 1995.
Epidemiologic and biochemical studies have suggested an anti-inflammatory effect of n-3 fatty acids. Beneficial therapeutic effects reported from small patient groups need to be confirmed in large-cohort controlled clinical trials. There is a growing number of clinical trials of n-3 fatty acid supplementation in disease. Clinical benefits have been moderate in patients with rheumatoid arthritis and with arterial hypertension. Clearly negative results have been reported during the past 2 years for patients with lupus nephritis and for patients with psoriasis or with atopic dermatitis. Such trials have now been completed. For patients with coronary artery disease following coronary angioplasty, earlier results of a large meta-analysis, could not be confirmed. For patients with IgA-nephropathy and for patients following kidney transplantation, a clear benefit was seen in patients receiving fish oil. These promising results are currently pursued in follow-up phase III clinical trials.


Correction of syndactyly: advantages with a non-grafting technique and the use of absorbable skin sutures.
Correction of syndactyly without skin grafts is simple and reliable. Combined with an absorbable suture technique the non-grafting method is further advantageous minimising risks and saving time and costs. Three methods are compared: skin grafts plus monofilament polyamide (Ethilon, Ethicon) (n = 32), no skin grafts plus Ethilon (n = 19), and no skin grafts plus polyglactin 910 (Vicryl rapide, Ethicon) (n = 9). The two groups in which grafting was not used had significantly shorter operations (mean 86 minutes compared with 118 minutes) and fewer complications. They also required fewer operations and spent less time in hospital.


Atomic-resolution imaging with a sub-50-pm electron probe.
Using a highly coherent focused electron probe in a fifth-order aberration-corrected transmission electron microscope, we report on resolving a crystal spacing less than 50 pm. Based on the geometrical source size and residual coherent and incoherent axial lens aberrations, an electron probe is calculated, which is theoretically capable of resolving an ideal 47 pm spacing with 29% contrast. Our experimental data show the 47 pm spacing of a Ge 114 crystal imaged with 11%-18% contrast at a 60%-95% confidence level, providing the first direct evidence for sub-50-pm resolution in annular dark-field scanning transmission electron microscopy imaging.


YY1 and CP2c in Unidirectional Spermatogenesis and Stemness.
Spermatogonial stem cells (SSCs) have stemness characteristics, including germ cell-specific imprints that allow them to form gametes. Spermatogenesis involves changes in gene expression such as a transition from expression of somatic to germ cell-specific genes, global repression of gene expression, meiotic sex chromosome inactivation, highly condensed packing of the nucleus with protamines, and morphogenesis. These step-by-step processes finally generate spermatozoa that are fertilization competent. Dynamic epigenetic modifications also confer totipotency to germ cells after fertilization. Primordial germ cells (PGCs) in embryos do not enter meiosis, remain in the proliferative stage, and are referred to as gonocytes, before entering quiescence. Gonocytes develop into SSCs at about 6 days after birth in rodents. Although chromatin structural modification by Polycomb is essential for gene silencing in mammals, and epigenetic changes are critical in spermatogenesis, a comprehensive understanding of transcriptional regulation is lacking. Recently, we evaluated the expression profiles of Yin Yang 1 (YY1) and CP2c in the gonads of E14.5 and 12-week-old mice. YY1 localizes at the nucleus and/or cytoplasm at specific stages of spermatogenesis, possibly by interaction with CP2c and YY1-interacting transcription factor. In the present article, we discuss the possible roles of YY1 and CP2c in spermatogenesis and stemness based on our results and a review of the relevant literature.


The cardiac sodium channel mutation delQKP 1507-1509 is associated with the expanding phenotypic spectrum of LQT3, conduction disorder, dilated cardiomyopathy, and high incidence of youth sudden death.
We report diverse phenotypic consequences of the delQKP-1507-1509 cardiac sodium channel mutation in three generations of a Chinese family. Clinical and electrocardiographic (ECG), echocardiographic examination was followed by direct sequencing of SCN5A, KCNQ1, HERG, and LAMIN A/C to screen genomic DNA from blood samples. Of two mutation carriers, the proband was born with conduction disorders including second-degree atrioventricular (AV) block with prolonged QTc interval, additionally showing left anterior fascicular block (LAFB), incomplete right bundle-branch block (IRBBB), and intermittent third-degree AV block at 2 years, and clinical presentations of multiple syncope despite normal electroencephalograms at 8 years. Continuous ECG monitoring following presentation at 13 years revealed prolonged QTc and biphasic T-waves, multiple episodes of ventricular tachycardia, ventricular fibrillation, and torsades de pointes. Transthoracal echocardiography then revealed left ventricular dilatation and reduced systolic function. Another mutation carrier showed features of long QT syndrome type 3 (LQT3), LAFB, and dilated cardiomyopathy (DCM). Two additional subjects died suddenly at 13 and 33 years. This data compliments and expands the spectrum of phenotypes resulting from this known gain-of-function mutation, including not only LQT3, cardiac conduction defects, and sudden death but also DCM, hitherto associated with loss-of-function mutations, for the first time.


Tongue- and Jaw-Specific Contributions to Acoustic Vowel Contrast Changes in the Diphthong /ai/ in Response to Slow, Loud, and Clear Speech.
This study sought to determine decoupled tongue and jaw displacement changes and their specific contributions to acoustic vowel contrast changes during slow, loud, and clear speech. Twenty typical talkers repeated "see a kite again" 5 times in 4 speech conditions (typical, slow, loud, clear). Speech kinematics were recorded using 3-dimensional electromagnetic articulography. Tongue composite displacement, decoupled tongue displacement, and jaw displacement during /ai/, as well as the distance between /a/ and /i/ in the F1-F2 vowel space, were examined during the diphthong /ai/ in "kite." Displacements significantly increased during all 3 speech modifications. However, jaw displacements increased significantly more during clear speech than during loud and slow speech, whereas decoupled tongue displacements increased significantly more during slow speech than during clear and loud speech. In addition, decoupled tongue displacements increased significantly more during clear speech than during loud speech. Increases in acoustic vowel contrast tended to be larger during slow speech than during clear speech and were predominantly tongue-driven, whereas those during clear speech were fairly equally accounted for by changes in decoupled tongue and jaw displacements. Increases in acoustic vowel contrast during loud speech were smallest and were predominantly tongue-driven, particularly in men. Findings suggest that task-specific patterns of decoupled tongue and jaw displacement change and task-specific patterns of decoupled tongue and jaw contributions to vowel acoustic change across these speech modifications. Clinical implications are discussed.


Something sinistral going on? Asymmetry in arm movement frequency.
In a test of the hypothesis that the dominant arm would be more active than the nondominant arm, 70 young adults (including 34 left-handers) wore instrumented motion recorders on both wrists for 48 hours. Participants' left arms made approximately 80 more movements per hour than did their right arms, which was unexpected. This mean difference reflected a generalised leftward shift in the distribution of the frequency of arm movements. Two-thirds of the individuals in the sample were characterised by a sinistral bias in arm movement frequencies. Degree of sinistral bias in arm movement frequency was unrelated to hand preference as measured by the Waterloo Inventory. Implications of this new asymmetry are discussed.


Adiponectin and resistin in PCOS: a clinical, biochemical and molecular genetic study.
We conducted a cross-sectional case-control study to evaluate the possible involvement of adiponectin and resistin in the pathogenesis of polycystic ovary syndrome (PCOS). Seventy-six PCOS patients and 40 non-hyperandrogenic women matched for BMI and degree of obesity were included. Serum adiponectin and resistin levels, anthropometrical and hormonal variables, the 45 T-->G and 276 G-->T polymorphisms in the adiponectin gene, and the -420 C-->G variant in the resistin gene, were analysed. Serum adiponectin concentrations were reduced in PCOS patients compared with controls (P = 0.038) irrespective of the degree of obesity, whereas serum resistin levels were increased in overweight and obese women compared with lean subjects (P = 0.016), irrespective of their PCOS or controls status. The adiponectin and resistin polymorphisms were not associated with PCOS and did not influence serum levels of adiponectin, resistin and other clinical and hormonal variables. In a multiple regression model, the waist-to-hip ratio, free testosterone levels and age, but not insulin resistance, were the major determinants of hypoadiponectinaemia. PCOS patients present with hypoadiponectinaemia, in relation with abdominal adiposity and hyperandrogenism. Our present results suggest that hyperandrogenism and abdominal obesity, by reducing the serum levels of the insulin sensitizer adipokine adiponectin, might contribute to the insulin resistance of PCOS.


MicroRNAs underlying memory deficits in neurodegenerative disorders.
Neurodegenerative disorders are defined by neuronal loss and often associated with dementia. Understanding the multifactorial nature of cognitive decline is of particular interest. Cell loss is certainly a possibility but also an early imbalance in the complex gene networks involved in learning and memory. The small (~22nt) non-coding microRNAs play a major role in gene expression regulation and have been linked to neuronal survival and cognition. Interestingly, changes in microRNA signatures are associated with neurodegenerative disorders. In this review, we explore the role of three microRNAs, namely miR-132, miR-124 and miR-34, which are dysregulated in major neurodegenerative diseases such as Alzheimer's disease, Parkinson's disease and Huntington's disease. Interestingly, these microRNAs have been associated with both memory impairment and neuronal survival, providing a potential common molecular mechanism contributing to dementia.


A Study on the Dietary Intake and the Nutritional Status among the Pancreatic Cancer Surgical Patients.
The adequate dietary intake is important to maintain the nutritional status of the patients after pancreatic cancer surgery. This prospective study was designed to investigate the dietary intake and the nutritional status of the patients who had pancreatic cancer surgery. Thirty-one patients (15 men, 16 women) were enrolled and measured body weight, body mass index (BMI), nutritional risk index (NRI), and Malnutrition Universal Screening Tool (MUST). Actual oral intake with nutritional impact symptoms recorded on the clinical research foam at every meal and medical information were collected from electronic medical charts. The rates of malnutrition at admission were 45.1% (14/31) and 28.9% (9/31) by NRI and MUST method, respectively, but those were increased to 87% (27/31) and 86.6% (26/31) after operation on discharge. The median values of daily intake of energy, carbohydrates, fat, and protein were 588.1 kcal, 96.0 g, 11.8 g, and 27.0 g, respectively. Most patients (n = 20, 64.5%) experienced two or more symptoms such as anorexia, abdominal bloating and early satiety. There were negative correlations between C-reactive protein (CRP) levels and the intake of total energy, protein, fat, and zinc. The rates of malnutrition were increased sharply after surgery and the dietary intake also influenced the inflammatory indicators. The results suggested that need of considering special therapeutic diets for the patients who received pancreatic surgery.


Genome-Wide Methylation Profiling in the Thalamus of Scrapie Sheep.
Scrapie is a neurodegenerative disorder belonging to the group of transmissible spongiform encephalopathy (TSE). Scrapie occurs in sheep and goats, which are considered good natural animal models of these TSE. Changes in DNA methylation occur in the central nervous system (CNS) of patients suffering from prion-like neurodegenerative diseases, such as Alzheimer's disease. Nevertheless, potential DNA methylation alterations have not yet been investigated in the CNS of any prion disease model or naturally infected cases, neither in humans nor in animals. Genome-wide DNA methylation patterns were studied in the thalamus obtained from sheep naturally infected with scrapie at a clinical stage (n = 4) and from controls (n = 4) by performing a whole-genome bisulfite sequencing (WGBS) analysis. Ewes carried the scrapie-susceptible ARQ/ARQ PRNP genotype and were sacrificed at a similar age (4-6 years). Although the average genomic methylation levels were similar between the control and the scrapie animals, we identified 8,907 significant differentially methylated regions (DMRs) and 39 promoters (DMPs). Gene Ontology analysis revealed that hypomethylated DMRs were enriched in genes involved in transmembrane transport and cell adhesion, whereas hypermethylated DMRs were related to intracellular signal transduction genes. Moreover, genes highly expressed in specific types of CNS cells and those previously described to be differentially expressed in scrapie brains contained DMRs. Finally, a quantitative PCR (qPCR) validation indicated differences in the expression of five genes (PCDH19, SNCG, WDR45B, PEX1, and CABIN1) that matched the methylation changes observed in the genomic study. Altogether, these results suggest a potential regulatory role of DNA methylation in prion neuropathology.


Physical exposures during pregnancy and congenital cardiovascular malformations.
Congenital cardiovascular malformations (CCM) cause substantial neonatal morbidity and mortality. Known risk factors for CCM explain only 10-20% of all cases. Few studies have examined mothers' physical exposures during pregnancy and the risk of CCM in their offspring. This study examined the association between exposures to extreme temperatures, prolonged standing, and heavy lifting during early pregnancy and risk of CCM in offspring. Using a case-control study design, 502 cases and 1066 controls were drawn from the population of all liveborn infants born between January 1988 and June 1991 to mothers living in 14 counties in New York State. Cases were identified from a population-based registry of congenital malformations. Controls were randomly selected from birth certificate records. Interviews were conducted by telephone, using a structured questionnaire. Exposure estimates were based on women's self-reports of conditions in the residence and workplace. Eighty-three per cent of the mothers were white, and 66% were between 25 and 34 years old. After adjusting all results for known risk factors and confounding variables, we found no significant increased risk of CCM in subjects whose mothers reported being exposed during early pregnancy to extreme heat (OR = 1.13, 95% CI 0.59, 2.19), nor to extreme cold (OR = 1.19, 95% CI 0.66, 2.15). Mothers who reported ever using a hot tub, hot bath, or sauna during early pregnancy had no increased risk of CCM in their offspring (OR = 0.88, 95% CI 0.65, 1.18). Performing heavy lifting during early pregnancy did not increase the risk of CCM in offspring (OR = 0.80, 95% CI 0.57, 1.11). Prolonged standing during early pregnancy was not associated with an increased risk of CCM in children (OR = 1.03, 95% CI 0.82, 1.28). Thus if these maternal exposures have an adverse effect, it is unlikely to involve CCMs.


cDNA cloning, heterologous expressions, and functional characterization of malonyl-coenzyme a:anthocyanidin 3-o-glucoside-6"-o-malonyltransferase from dahlia flowers.
In the flowers of important ornamental Compositae plants, anthocyanins generally carry malonyl group(s) at their 3-glucosyl moiety. In this study, for the first time to our knowledge, we have identified a cDNA coding for this 3-glucoside-specific malonyltransferase for anthocyanins, i.e. malonyl-coenzyme A:anthocyanidin 3-O-glucoside-6"-O-malonyltransferase, from dahlia (Dahlia variabilis) flowers. We isolated a full-length cDNA (Dv3MaT) on the basis of amino acid sequences specifically conserved among anthocyanin acyltransferases of the versatile plant acyltransferase family. Dv3MaT coded for a protein of 460 amino acids. Quantitative real-time PCR analyses of Dv3MaT showed that the transcript was present in accordance with the distribution of 3MaT activities and the anthocyanin accumulation pattern in the dahlia plant. The Dv3MaT cDNA was expressed in Escherichia coli, and the recombinant enzyme was purified to homogeneity and characterized. The recombinant Dv3MaT catalyzed the regiospecific transfer of the malonyl group from malonyl-coenzyme A (K(m), 18.8 microM) to pelargonidin 3-O-glucoside (K(m), 46.7 microM) to produce pelargonidin 3-O-6"-O-malonylglucoside with a k(cat) value of 7.3 s(-1). The other enzymatic profiles of the recombinant Dv3MaT were closely related to those of native anthocyanin malonyltransferase activity in the extracts of dahlia flowers. Dv3MaT cDNA was introduced into petunia (Petunia hybrida) plants whose red floral color is exclusively provided by cyanidin 3-O-glucoside and 3,5-O-diglucoside. Thirteen transgenic lines of petunia were found to produce malonylated products of these anthocyanins (11-63 mol % of total anthocyanins in the flower). The spectral stability of cyanidin 3-O-6"-O-malonylglucoside at the pHs of intracellular milieus of flowers was significantly higher than that of cyanidin 3-O-glucoside. Moreover, 6"-O-malonylation of cyanidin 3-O-glucoside effectively prevented the anthocyanin from attack of beta-glucosidase. These results suggest that malonylation should serve as a strategy for pigment stabilization in the flowers.


Surfactant wound irrigation for the treatment of staphylococcal clinical isolates.
Deep wound infection involving an implanted biomaterial is a devastating complication in orthopaedic surgery. Two-thirds of such infections are monomicrobial and the most commonly isolated bacteria in human osteomyelitis and orthopaedic device infection are Staphylococcus aureus and Staphylococcus epidermidis. The purpose of the current study was to examine the effectiveness of the previously reported sequential surfactant irrigation protocol against human-isolated clinical strains of Staphylococcus aureus and Staphylococcus epidermidis in the rat model of orthopaedic implant contamination. The infectivity rate of human-isolated clinical strains of Staphylococcus aureus in a contaminated complex orthopaedic wound was reduced effectively by a sequential surfactant irrigation protocol. Also, in this model, the infectivity of Staphylococcus epidermidis was reduced by normal saline irrigation alone when compared with no irrigation. Therefore, the sequential surfactant irrigation protocol may represent an effective method of wound irrigation in monomicrobial Staphylococcus aureus orthopaedic implant contamination, and normal saline irrigation may suffice in cases of monomicrobial Staphylococcus epidermidis contamination. Additional studies are necessary to determine the clinical use of surfactant irrigation.


Modeling the intrinsic dynamics of foot-and-mouth disease.
We propose a new mathematical modeling framework to investigate the transmission and spread of foot-and-mouth disease. Our models incorporate relevant biological and ecological factors, vaccination effects, and seasonal impacts during the complex interaction among susceptible, vaccinated, exposed, infected, carrier, and recovered animals. We conduct both epidemic and endemic analysis, with a focus on the threshold dynamics characterized by the basic reproduction numbers. In addition, numerical simulation results are presented to demonstrate the analytical findings.