High spatiotemporal resolution imaging of fast intrinsic optical signals activated by retinal flicker stimulation.
High resolution monitoring of stimulus-evoked retinal neural activities is important for understanding retinal neural mechanisms, and can be a powerful tool for retinal disease diagnosis and treatment outcome evaluation. Fast intrinsic optical signals (IOSs), which have the time courses comparable to that of electrophysiological activities in the retina, hold the promise for high resolution imaging of retinal neural activities. However, application of fast IOS imaging has been hindered by the contamination of slow, high magnitude optical responses associated with transient hemodynamic and metabolic changes. In this paper we demonstrate the feasibility of separating fast retinal IOSs from slow optical responses by combining flicker stimulation and dynamic (temporal) differential image processing. A near infrared flood-illumination microscope equipped with a high-speed (1000 Hz) digital camera was used to conduct concurrent optical imaging and ERG measurement of isolated frog retinas. High spatiotemporal resolution imaging revealed that fast IOSs could follow flicker frequency up to at least 6 Hz. Comparable time courses of fast IOSs and ERG kinetics provide evidence that fast IOSs are originated from stimulus activated retinal neurons.


A transactional analysis of the relation between maternal sensitivity and child vagal regulation.
A transactional model examining the longitudinal association between vagal regulation (as indexed by vagal withdrawal) and maternal sensitivity from age 2.5 to age 5.5 was assessed. The sample included 356 children (171 male, 185 female) and their mothers who participated in a laboratory visit at age 2.5, 4.5, and 5.5. Cardiac vagal tone was obtained during a baseline task and during emotional frustration tasks. Maternal sensitivity was assessed via direct observation during a pretend play and cleanup task. To test for transactional associations, a path model estimating stability paths for vagal withdrawal and maternal sensitivity was compared with a full reciprocal model that included all cross-lagged pathways. A chi-square difference test was used to evaluate whether the cross-lagged model explained the data above and beyond the stability model. The vagal withdrawal cross-lagged model was found to fit significantly better than the stability model and revealed that maternal sensitivity at 2.5 years was associated positively with vagal withdrawal at 4.5 years, and vagal withdrawal at 4.5 years was associated positively with maternal sensitivity at 5.5 years. These results suggest that early sensitive responding by mothers was associated with increases in vagal withdrawal, which in turn was associated with higher levels of sensitive parenting.


Autoregulation of rat pituitary prolactin secretion demonstrated by a new perfusion method.
Regulation of prolactin secretion was investigated by perfusing rat pituitaries in vitro. Two pituitary glands from inbred rats were transplanted beneath the renal capsule of a third recipient rat. Three weeks later, the transplanted kidney was removed and perfused in vitro with a defined cell-free medium. Normal renal function was maintained during perfusion, and cell morphology of the transplants remained unchanged as assessed by electron microscopy. Pituitary prolactin content did not change after 120 min of perfusion despite release of approximately 10 micrograms of hormone. Thyrotropin-releasing hormone (10 ng/ml) did not stimulate prolactin release; dopamine (20 ng/ml) rapidly, but transiently inhibited prolactin release; bromocriptine (20 ng/ml) rapidly and persistently inhibited prolactin release; haloperidol (100 ng/ml) blocked the inhibition by dopamine or bromocriptine, but when given alone inhibited prolactin release. Finally, prolactin release was also inhibited by the presence of 100 and 200 ng/ml, but not 50 ng/ml of NIAMDD RP-1 rat prolactin. It is concluded that in vitro perfusion of transplanted rat pituitaries provides a new model for studying the direct effect of agents on the secretion of prolactin from the pituitary and that rat prolactin and/or its metabolites directly inhibit pituitary prolactin secretion.


PcpA of Streptococcus pneumoniae mediates adherence to nasopharyngeal and lung epithelial cells and elicits functional antibodies in humans.
Streptococcus pneumoniae (pneumococci) adhere to human nasopharyngeal (NP) epithelial cells as a first step in colonization and adherence of pneumococci to lung epithelia may be required to establish pneumonia. We sought to determine if PcpA can serve as an adhesin to human NP (D562) and lung (A549) epithelial cells and whether PcpA mediated adherence can be inhibited by human anti-PcpA antibodies. A PcpA isogenic mutant was constructed in a pneumococcal TIGR4 background. When the mutant and wild type strains were compared for their adherence to D562 and A549 cell lines, a reduction in adherence by the mutant was observed (p = 0.0001 for both cell types). PcpA was ectopically expressed on the surface of minimally-adherent heterologous host Escherichia coli resulting in augmented adherence to D562 (p = 0.002) and A549 (p = 0.015) cells. Total IgG was purified from a pool of 6 human sera having high IgG titers of anti-pneumococcal proteins. The purified IgG reduced TIGR4 adherence to D562 cells but we determined that this effect was largely due to bacterial cell aggregation as determined by flow cytometry and confocal microscopy. Fab fragments were prepared from pooled IgG sera. Inhibition of TIGR4 adherence to D562 cells was observed using the Fab fragments without causing bacterial aggregation (p = 0.0001). Depletion of PcpA-specific Fab fragments resulted in an increase in adherence of TIGR4 to D562 cells (p = 0.028). We conclude that PcpA can mediate adherence of pneumococci to human NP and lung epithelial cells and PcpA mediated adherence can be inhibited by human anti-PcpA antibodies.


Thermally drawn fibers as nerve guidance scaffolds.
Synthetic neural scaffolds hold promise to eventually replace nerve autografts for tissue repair following peripheral nerve injury. Despite substantial evidence for the influence of scaffold geometry and dimensions on the rate of axonal growth, systematic evaluation of these parameters remains a challenge due to limitations in materials processing. We have employed fiber drawing to engineer a wide spectrum of polymer-based neural scaffolds with varied geometries and core sizes. Using isolated whole dorsal root ganglia as an in vitro model system we have identified key features enhancing nerve growth within these fiber scaffolds. Our approach enabled straightforward integration of microscopic topography at the scale of nerve fascicles within the scaffold cores, which led to accelerated Schwann cell migration, as well as neurite growth and alignment. Our findings indicate that fiber drawing provides a scalable and versatile strategy for producing nerve guidance channels capable of controlling direction and accelerating the rate of axonal growth.


A psychological profile of the transsexual. II. The female.
This research introduced standardized psychological measurement into the clinical assessment of the female transsexual. Twenty females with a presenting complaint of gender dysphoria were carefully screened to ensure high correspondence between their clinical pictures and current nosological concepts of transsexualism. All were administered the Derogatis Sexual Functioning Inventory as a part of their initial clinical work-up, and their profiles were contrasted with those of a comparison group of 143 heterosexual females. Results indicated hypermasculine gender role definitions and fantasy themes among the transsexual group with decrements in terms of sexual information and range of sexual experiences. The transsexual females revealed moderately dysphoric affect profiles but were unremarkable in terms of psychological symptoms. Overall, there was evidence of reductions in sexual drive; however, this result was highly dependent upon which indicator of drive was employed. The opportunity to obtain a systematic psychometric evaluation of this fascinating clinical group is viewed as a significant step in gaining an ultimate understanding of their condition.


Prostate cancer and bone metastases: medical treatment.
Prostate cancer is the most common malignancy in men in the United States. With the long natural history of the disease, management of skeletal morbidity related to advanced prostate cancer becomes a major public health issue. The standard of care in advanced prostate cancer is androgen deprivation therapy. This may accelerate the development of osteoporosis and further exacerbate the risks of having adverse skeletal-related events develop. Recently, the use of bisphosphonates in men who have not responded to androgen deprivation therapy has been shown to reduce the incidence of skeletal-related events with time. Questions remain as to whether bisphosphonates should be broadly applied to earlier stages of the disease or tailored to men at higher risk of having bone-related morbidity. Work is ongoing to improve other approaches to the medical treatment of bone metastases in patients with advanced prostate cancer including the use of radiopharmaceuticals and combined chemotherapy.


Poly(ethylene oxide)-block-polyphosphester-based Paclitaxel Conjugates as a Platform for Ultra-high Paclitaxel-loaded Multifunctional Nanoparticles.
A new type of degradable, nanoscopic polymer assembly containing ultra-high levels of drug loading via covalent attachment within amphiphilic core-shell nanoparticle morphology has been generated as a potentially effective and safe anti-cancer agent. Poly(ethylene oxide)-block-polyphosphoester-based paclitaxel drug conjugates (PEO-b-PPE-g-PTX) were synthesized by rapid, scalable and versatile approach that involves only two steps: organocatalyst-promoted ring-opening-polymerization followed by click reaction-based conjugation of a PTX prodrug. Variations in the polymer-to-PTX stoichiometries allowed for optimization of the conjugation efficiency, the PTX drug loading and the resulting water solubilities of the entire polymer and the PTX content. The PEO-b-PPE-g-PTX formed well-defined micelles in aqueous solution, with a PTX loading capacity as high as 65 wt%, and a maximum PTX concentration of 6.2 mg/mL in water, which is 25000-fold higher than the aqueous solubility of free PTX. The positive cell-killing activity of PEO-b-PPE-g-PTX against several cancer cell lines is demonstrated, and the presence of pendant reactive functionality provides a powerful platform for future work to involve conjugation of multiple drugs and imaging agents to achieve chemotherapy and bioimaging.


Stimulus-specific alteration of the relationship between cytosolic Ca(2+) transients and nitric oxide production in endothelial cells ex vivo.
1. To investigate the quantitative relationship between elevation in the intracellular Ca(2+) concentration ([Ca(2+)](i)) and nitric oxide (NO) production, the changes in [Ca(2+)](i) and NO production were determined in parallel, using fluorimetry of fura-2 and 2, 3-diaminonaphthalene, respectively, in endothelial cells ex vivo of pig aortic valves. 2. The extent of [Ca(2+)](i) elevation was quantitatively assessed by two parameters: the level of peak [Ca(2+)](i) elevation and the area under the [Ca(2+)](i) curve during treatment (the integrated [Ca(2+)](i) elevation). The amount of NO production was expressed as a percentage of that obtained with 10 microM ATP for 3 min. 3. ATP, bradykinin, thrombin, and ionomycin were used as stimulation to induce NO production, and all these caused [Ca(2+)](i) increases and NO production in a concentration-dependent manner. 4. The relationships between the peak [Ca(2+)](i) and NO production or between the integrated [Ca(2+)](i) elevation and NO production were well described by a straight line. However, the slope value of the linear relationship in both cases varied with the type of stimulation, with thrombin giving the greatest value, followed by ATP, bradykinin and ionomycin. 5. These data suggest that in endothelial cells ex vivo: (1) [Ca(2+)](i) elevation regulates NO production, but (2) the peak [Ca(2+)](i) elevation- or the integrated [Ca(2+)](i) elevation-NO production relationships varies depending on the type of agonists. Our results thus demonstrate the presence of the agonists-dependent modulation of the relationship between [Ca(2+)](i) elevation and NO production in endothelial cells ex vivo.


EFFECT OF PERILLA FRUTESCENS EXTRACTS AND ROSMARINIC ACID ON RAT HEART MITOCHONDRIAL FUNCTIONS.
Perilla frutescens L. due to its aromatic, antibacterial, anti-inflammatory and antioxidant traits has been traditionally used as medicinal plant in Eastern Asia. Alterations of mitochondria are interconnected with many chronic diseases. Bioactives of herbal extracts can modulate mitochondrial effects and be beneficial in prevention of mitochondrial related chronic diseases. Direct effects of the red-leaf form P. frutescens extract (PFE) and the green-leaf form P. frutescens var. crispa f. viridis extract (PCE) were evaluated investigating activities on the oxidative phosphorylation and antioxidant activity in the rat heart mitochondria in vitro. HPLC-MS analysis was applied for the identification of phenolic compounds. Cell with a Clark-type oxygen electrode was used for mitochondrial respiration measurement. The generation of reactive oxygen species was estimated in isolated rat heart mitochondria and determined fluorimetrically. State 3 respiration rate was not affected by lower concentrations, however, it was inhibited at higher concentrations by 22-70% for PFE and by 45-55% for PCE. PFE containing anthocyanins induced the concentration-dependent stimulation (by 23-76%) of the State 4 respiration rate after addition of cytochrome c due to reducing properties. Significant reduction of H₂O₂ pro- duction was observed with investigated concentrations of rosmarinic acid and both perilla extracts. Our results demonstrate that the effect of PFE and PCE extracts on rat heart mitochondria depend on the qualitative characteristics of complex of biologically active compounds. Selective effects on mitochondrial function could enable the regulation of apoptosis or another mechanisms occurring in cells.


Risks for possible and probable opioid misuse among recipients of chronic opioid therapy in commercial and medicaid insurance plans: The TROUP Study.
The use of chronic opioid therapy (COT) for chronic non-cancer pain (CNCP) has increased dramatically in the past two decades. There has also been a marked increase in the abuse of prescribed opioids and in accidental opioid overdose. Misuse of prescribed opioids may link these trends, but has thus far only been studied in small clinical samples. We therefore sought to validate an administrative indicator of opioid misuse among large samples of recipients of COT and determine the demographic, clinical, and pharmacological risks associated with possible and probable opioid misuse. A total of 21,685 enrollees in commercial insurance plans and 10,159 in Arkansas Medicaid who had at least 90 days of continuous opioid use 2000-2005 were studied for one year. Criteria were developed for possible and probable opioid misuse using administrative claims data concerning excess days supplied of short-acting and long-acting opioids, opioid prescribers and opioid pharmacies. We estimated possible misuse at 24% of COT recipients in the commercially insured sample and 20% in the Medicaid sample and probable misuse at 6% in commercially insured and at 3% in Medicaid. Among non-modifiable factors, younger age, back pain, multiple pain complaints and substance abuse disorders identify patients at high risk for misuse. Among modifiable factors, treatment with high daily dose opioids (especially >120 mg MED per day) and short-acting Schedule II opioids appears to increase the risk of misuse. The consistency of the findings across diverse patient populations and the varying levels of misuse suggest that these results will generalize broadly, but await confirmation in other studies.


Fine structure and chemical analysis of the metathoracic scent gland secretion in Graphosoma lineatum (Linnaeus, 1758) (Heteroptera, Pentatomidae).
Morphology and ultrastructure of metathoracic scent glands (MTGs) of Graphosoma lineatum (Heteroptera, Pentatomidae) were studied by scanning electron microscopy (SEM) and transmission electron microscopy (TEM). Extracts of the volatile fraction of the MTG secretion from males and females were subjected to initial analysis. One pair of the MTG is composed of a reservoir and a pair of lateral glands connected to the reservoir with a duct. MTGs are open in between the meso- and metacoxae, on evaporation areas with a mushroom-like structure. Reservoir walls embody two types of cells, type I and type II, respectively. Cells of type I have numerous organelles, while type II cells have only been found in a certain area of the reservoir wall. They have large secretory ducts lined by a cuticular intima layer. The lateral glands connected to reservoir have two further types of cells. Lateral glands are lined by type A secretory cells and secretory duct is found in their cytoplasm. Type B cells are poor in organelles and are smaller than type A cells. Coupled gas chromatography-mass spectrometry examinations revealed that both in males and females of G. lineatum MTGs 14 chemical compounds occur, among which only 10 are common. These secretions indicate information such as defensive and pheromonal activities, other functions could be attributed to the secretion of the MTG of G. lineatum by comparison with other closely related bug species. In the analyses of MTGs of females of G. lineatum, n-octadecanoic acid was observed at the most and n-undecane was determined at lowest level, while males of G. lineatum n-tridecane was determined at the highest level; and (E)-2-hexenyl acetate has been observed at the lowest.


Psychostimulant-induced Fos protein expression in the thalamic paraventricular nucleus.
Lesions of glutamatergic afferents to the nucleus accumbens have been reported to block psychostimulant-induced behavioral sensitization. However, thalamic glutamatergic projections to the nucleus accumbens have received little attention in the context of psychostimulant actions. We examined the effects of acute amphetamine and cocaine administration on expression of Fos protein in the thalamic paraventricular nucleus (PVT), which provides glutamatergic inputs to the nucleus accumbens and also receives dopaminergic afferents. Immunoblot and immunohistochemical studies revealed that both psychostimulants dose-dependently increased PVT Fos expression. PVT neurons retrogradely labeled from the nucleus accumbens were among the PVT cells that showed a Fos response to amphetamine. D2 family dopamine agonists, including low doses of the D3-preferring agonist 7-OH-DPAT, increased the numbers of Fos-like-immunoreactive neurons in the PVT. Conversely, the effects of cocaine and amphetamine on PVT Fos expression were blocked by pretreatment with the dopamine D2/3 antagonist raclopride. Because PVT neurons express D3 but not other dopamine receptor transcripts, it appears that psychostimulants induce Fos in PVT neurons through a D3 dopamine receptor. We suggest that the PVT may be an important part of an extended circuit subserving both the arousing properties and reinforcing aspects of psychostimulants.


Prospective association between a dietary quality index based on a nutrient profiling system and cardiovascular disease risk.
Public health strategies are essential to guide consumers' choices and produce a substantial population impact on cardiovascular disease risk prevention through nutrition. Our aim was to investigate the prospective association between the Food Standards Agency nutrient profiling system dietary index (FSA-NPS DI) and cardiovascular disease risk. The FSA-NPS has been proposed to serve as a basis for a five-colour nutrition label suggested in France to be put on the front of pack of food products. A total of 6515 participants to the SU.VI.MAX cohort (1994-2007), who completed at least six 24-hour dietary records during the first two years of the study, were followed for a median of 12.4 years (25th-75th percentiles: 11.0-12.6). Multivariable Cox proportional hazards models were used to characterise the associations between FSA-NPS DI (continuous and sex-specific quartiles) and the incidence of cardiovascular diseases. Interactions with individual characteristics were tested; 181 major cardiovascular events were reported (59 myocardial infarctions, 43 strokes, 79 anginas). A higher FSA-NPS DI, characterising poorer food choices, was associated with an overall increase in cardiovascular disease risk (HRfor a 1-point increment = 1.14 (1.03-1.27); HRQ4vs.Q1 = 1.61 (1.05-2.47), Ptrend Q4-Q1 = 0.03). This association tended to be stronger in smokers (HRfor a 1-point increment = 1.39 (1.11-1.73); Pinteraction = 0.01) and those less physically active (HRfor a 1-point increment = 1.26 (1.08-1.46); Pinteraction = 0.04). Our results suggest that poorer food choices, as reflected by a higher FSA-NPS DI, may be associated with a significant increase in cardiovascular risk, especially in at-risk individuals (smokers and physically inactive persons). This score could be a useful tool for public health prevention strategies. ClinicalTrials.gov identifier: NCT00272428.


Motor unit discharge characteristics and short term synchrony in paraplegic humans.
Frequency of firing and regularity of discharge of human motor units, and short term synchrony between pairs of motor units, have been assessed in extensor digitorum communis (EDC) and tibialis anterior (TA) muscles in control subjects and in clinically complete paraplegic subjects. The discharge pattern of TA motor units in paraplegia ranged from extremely regular to very irregular for different motor units whereas in the control population, and in EDC of both groups, there was a narrow, but intermediate, range of regularity. There was little difference in the incidence and degree of short term synchrony (STS) in EDC between paraplegic and normal subjects. In contrast, virtually no STS of motor units was observed in the TA muscles of the paraplegic group whereas control subjects exhibited approximately the same amount of STS in their TA and EDC muscles. It is concluded that the extra burden placed on arm muscles in paraplegia does not change the amount of synchronisation between motor units. Furthermore, section of the spinal cord does not increase STS as predicted from lesions of the reticulospinal tract in cats. This may reflect the coincidental removal of supraspinal synchronising inputs of motoneurons or the reorganisation of synaptic inputs in chronic paraplegia.


[Benign phylloides tumor of the breast. Considerations on a clinical case].
Phyllodes tumors are unusual biphasic fibroepithelial neoplasms of the breast, accounting for < 1% of all breast tumors and raising issues of diagnosis and therapeutic choice. They can grow quickly and when the maximum diameter is greater than 10 cm. We talk about giant phyllodes tumors. Ultrasound, Mammography and FNA are not effective. A potentially useful diagnostic modality is MRI. Core tissue biopsy or incisional biopsy represent the preferred means of pre-operative diagnosis. Conservative treatment can be effective also in giant tumors depending upon the size of the tumor and the breast if a complete excision with an adequate margin of normal breast tissue can be achieved, so avoiding local recurrence often accompanied by worse histopathology. The Authors report the case of a giant benign phyllode tumor of the breast treated with conservative surgery, quadrantectomy and oncoplasty. No local recurrence at 4 years follow-up.


Posttranslational modifications of brain and erythrocyte band 3 during aging and disease.
Band 3 performs the same structural and functional activities in adult brain as it does in erythrocytes. It ages as cells and tissues age Our studies, to date, indicate that the anion transport ability of band 3 decreases in brains and lymphocytes from old mice. This decreased transport ability precedes obvious structural changes, such as band 3 degradation and generation of SCA and is the earliest change thus far detected in band 3. The following changes occur in lymphocytes, erythrocytes and brain band 3 with aging: 1) a decreased efficiency of anion transport (decreased Vmax) in spite of an increase in number of anion binding sites (increased K(m)), 2) a decreased glucose transport, 3) 32P labeling in vitro, 4) an increased degradation to smaller fragments as detected by quantitative binding of antibodies to band 3 breakdown products and residue 812-830, and 5) a binding of physiologic IgG autoantibodies in situ. The latter three findings indicate that posttranslational changes occur. In addition, the anion transporter, band 3, undergoes an as yet undefined change that results in binding of "980" antibodies to aged band 3. Posttranslational changes in AD include decreased brain and RBC phosphorylation of a M(r) = 135, 113 and 45 kDa band 3 polypeptides due to the phosphorylation site being already occupied, increased degradation of band 3, alterations in band 3 recognized by antibodies, and decreased anion and glucose transport by blood cells. Band 3 in erythrocytes of AD patients has a different immunological identity from normal band 3 as evidenced by the binding of antibodies described in this study. AD may be preferentially manifested in the brain because neurons accumulate damage throughout the lifetime as they do not regenerate or undergo cell division. We suspect, and our data indicate, that the same mechanism(s) of AD occurs in all cells, but that the manifestations differ due to different cell proteins and functions.


Survival rate of salmonella on cooked pig ear pet treats at refrigerated and ambient temperature storage.
Pet treats, including pig ears, have been implicated as vehicles of human salmonellosis, and Salmonella has been isolated on commercially produced pig ears. Therefore, behavior of the pathogen on this very low water activity (aw) pet treat is of interest. The survival of Salmonella serotypes Newport and Typhimurium DT104 was measured on natural (aw 0.256) and smoked (aw 0.306) pig ear pet treat products inoculated at ca. 6.5 log CFU per sample and stored at 4.4 or 22°C for 365 days. Surviving populations of Salmonella were enumerated periodically, and a modified Weibull model was used to fit the inactivation curves for log populations. After 14 days, the decline of Salmonella was significantly (P < 0.05) greater at 22°C than at 4.4°C. By 365 days of storage at 4.4°C, Salmonella Typhimurium DT104 declined by 2.19 log on smoked pig ears and 1.14 log on natural pig ears, while Salmonella Newport declined by 4.20 log on smoked pig ears and 2.08 log on natural pig ears. Populations of Salmonella Typhimurium DT104 on refrigerated natural pig ears rebounded between day 152 (3.21 log CFU per sample) and day 175 (4.79 log CFU per sample) and rose gradually for the duration of the study to 5.28 log CFU per sample. The model fits for survival rate of Salmonella on pig ears at 4.4°C show a rapid initial decline followed by a long tailing effect. Salmonella Typhimurium DT104 on natural pig ears at 4.4°C had the slowest rate of reduction. At 22°C Salmonella declined nonlinearly by >4.5 log for each combination of serotype and pig ear type at 22°C but remained detectable by enrichment. The model parameter for days to first decimal reduction of Salmonella on pig ears was two to three times higher at 4.4°C compared with 22°C, demonstrating that Salmonella slowly declines on very low aw refrigerated pet treats and more rapidly at room temperature. This information may be useful for pet treat safety assessments.


Spontaneous variability in ventricular ectopic activity during chronic antiarrhythmic therapy.
Previous determinations of variability in frequency of ventricular arrhythmias have been based on repeated recordings obtained in the absence of therapy. We evaluate variability during "effective" treatment with antiarrhythmic drugs. Variability in the percent suppression of premature ventricular complexes (PVCs) was determined in 55 patients with chronic arrhythmias who underwent multiple ambulatory electrocardiographic recordings during evaluation of chronic therapy with antiarrhythmic drugs initially determined to be effective, which was defined as 70% or more reduction in total PVC frequency or 90% or more reduction in repetitive forms. During chronic therapy, total PVCs were suppressed by 92%, averaged after a logarithmic transformation step, and repetitive beats were suppressed by 88%. Variability in suppression was substantial. The one-sided 95% confidence intervals required a fall in suppression of total PVCs to 40% or less to exceed limits of spontaneous variability and of repetitive PVCs to 66% or less. Suppression declined at least once during therapy to less than 60% for total PVCs in 24 of 55 patients (44%) and to less than 80% for repetitive PVCs in 13 of 33 patients (39%); nine patients (16%) showed increases in PVC frequency at least once to levels above pretreatment baseline. Seven subgroups were analyzed for their effects on variability and loss of suppression: age, gender, disease etiology, cardiac function, baseline PVC frequency, use of beta-blockers, and class of antiarrhythmic drug. Differences in confidence bounds and loss of suppression were found to be determined in a complex way by subgroup differences in variability and in initial levels of PVC suppression. Variability was greater for patient subgroups with greater PVC frequency, beta-blocker therapy, and non-coronary artery disease. However, clinical loss of suppression was more common only in more elderly patients and those with worse cardiac function. In summary, substantial variability in arrhythmia frequency occurs during effective antiarrhythmic therapy, and the 95% confidence limits of spontaneous variability are broad and determined in a complex way. Careful consideration should be given before concluding on the basis of a single Holter test that changes (increases) in arrhythmia frequency, especially in certain subgroups, are caused by treatment failure.


Emotional Impact of COVID-19 Lockdown Among the Spanish Population.
The COVID-19 pandemic has resulted in some populations being confined to their homes as part of infection control measures. This situation can be hard to cope with due to separation from loved ones, prohibition of regular activities, fear of infection, loss of freedom, and so on. These negative impacts cause considerable psychological stress, and all the more so when the situation continues for an extended period, as was the case in Spain. The present study was aimed at investigating the effects of COVID-19 quarantine on the emotional functioning of confined Spanish individuals after 8 weeks of lockdown by means of a cross-sectional study. The possible associations between changes in emotional functioning and demographic variables (age and sex), health habits (physical exercise, following a routine, and smoking), social support, and resilience were also analyzed. A total of 906 Spanish adults completed an online survey to gather information about their prevailing mood and affects (before and after 8 weeks of lockdown), using the Positive and Negative Affect Schedule (PANAS) (Watson et al., 1988), and other variables related to their habits and protective factors. As expected, the data indicated an increase in negative affects (e.g., "upset," "afraid," "distressed") and a decrease in positive affects after 8 weeks under lockdown, as well as a general decline in overall mood. The largest increases in negative affects were observed in young adults (18-35 years) and women. We did not find any differences between people who were or were not diagnosed with COVID-19. Adhering to a routine, maintaining the same weight, and moderate physical exercise were associated with fewer negative affects, which indicates they are important protective factors, as are perceived social support and resilience. In order to mitigate the psychological impact of confinement, it is important to develop psychoeducational measures that encourage subjects to adhere to health habits and promote social support and resilience as protective factors. A special preventive focus should be placed on the most vulnerable population groups, namely women and young adults. For a public health lockdown to succeed, its negative consequences must be minimized insofar as possible through adequate knowledge of the risk factors and protective factors, and by means of prevention-oriented organization.


Photoionization-induced π↔ H site switching dynamics in phenol(+)-Rg (Rg = Ar, Kr) dimers probed by picosecond time-resolved infrared spectroscopy.
The ionization-induced π↔ H site switching reaction in phenol(+)-Rg (PhOH(+)-Rg) dimers with Rg = Ar and Kr is traced in real time by picosecond time-resolved infrared (ps-TRIR) spectroscopy. The ps-TRIR spectra show the prompt appearance of the non-vanishing free OH stretching band upon resonant photoionization of the π-bound neutral clusters, and the delayed appearance of the hydrogen-bonded (H-bonded) OH stretching band. This result directly proves that the Rg ligand switches from the π-bound site on the aromatic ring to the H-bonded site at the OH group by ionization. The subsequent H →π back reaction converges the dimer to a π↔ H equilibrium. This result is in sharp contrast to the single-step π→ H forward reaction in the PhOH(+)-Ar2 trimer with 100% yield. The reaction mechanism and yield strongly depend on intracluster vibrational energy redistribution. A classical rate equation analysis for the time evolutions of the band intensities of the two vibrations results in similar estimates for the time constants of the π→ H forward reaction of τ+ = 122 and 73 ps and the H →π back reaction of τ- = 155 and 188 ps for PhOH(+)-Ar and PhOH(+)-Kr, respectively. The one order of magnitude slower time constant in comparison to the PhOH(+)-Ar2 trimer (τ+ = 7 ps) is attributed to the decrease in density of states due to the absence of the second Ar in the dimer. The similar time constants for both PhOH(+)-Rg dimers are well rationalized by a classical interpretation based on the comparable potential energy surfaces, reaction pathways, and density of states arising from their similar intermolecular vibrational frequencies.


Oral feeding containers and their influence on intake and ventilation in preterm infants.
In order to determine whether changes in feeding container properties could expedite oral feeding without compromising ventilation, the rate of feeding and respiration were studied in 10 healthy preterm infants while using a collapsible feeding container, and the findings were compared to those obtained with the standard rigid bottle. Equal volumes of formula were offered from both containers. With the collapsible container, the total duration of feeding was significantly shorter, and the rate of ingestion of formula was significantly faster (p less than 0.01), while minute ventilation remained equally reduced from control levels during both feeds (p less than 0.05). The fall in ventilation was secondary to a reduction in tidal volume (p less than 0.001). Breathing frequency and transcutaneous oxygen tension did not change significantly with either trial. During feeding activity in both trials, airflow interruption occurred in both phases of the breathing cycle, but the total duration of interrupted airflow was greater with the collapsible container feed (p less than 0.001). Similar amounts of intra-oral negative pressure changes developed with sucking during both feeds. Results show that decreasing the rigidity of the feeding container shortened feeding time significantly without significantly affecting ventilation. Despite the greater duration of airflow interruption with the collapsible container, minute ventilation was sufficiently maintained to prevent compromised oxygenation.


A ketosynthase homolog uses malonyl units to form esters in cervimycin biosynthesis.
Ketosynthases produce the carbon backbones of a vast number of biologically active polyketides by catalyzing Claisen condensations of activated acyl and malonyl building blocks. Here we report that a ketosynthase homolog from Streptomyces tendae, CerJ, unexpectedly forms malonyl esters during the biosynthesis of cervimycin, a glycoside antibiotic against methicillin-resistant Staphylococcus aureus (MRSA). Deletion of cerJ yielded a substantially more active cervimycin variant lacking the malonyl side chain, and in vitro biotransformations revealed that CerJ is capable of transferring malonyl, methylmalonyl and dimethylmalonyl units onto the glycoside. According to phylogenetic analyses and elucidation of the crystal structure, CerJ is functionally and structurally positioned between the ketosynthase catalyzing Claisen condensations and acyl-ACP shuttles, and it features a noncanonical catalytic triad. Site-directed mutagenesis and structures of CerJ in complex with substrates not only allowed us to establish a model for the reaction mechanism but also provided insights into the evolution of this important subclass of the thiolase superfamily.


Intercostal Nerve Block and Neurolysis for Intractable Cancer Pain.
Management of intractable cancer-associated chest wall pain is difficult once patients have reached dose-limiting side effects of opioids and coanalgesic medications. This case series describes 11 patients with intractable cancer-associated chest wall pain who were treated with a diagnostic intercostal nerve block. Six patients subsequently received chemical neurolysis with phenol using the same approach. No serious adverse events were observed. Radiopaque contrast dye spread into the paravertebral space in all 11 patients, and in 1 patient contrast dye spread into the epidural space. Seven of 11 patients experienced pain relief from the diagnostic blockade. Four of six patients experienced pain relief from the neurolytic blockade. The principal reportable finding from this case series is the observation that contrast dye spread liberally from the intercostal space into other anatomic spaces, even though very small volumes of injectate (less than 5 mL) were used. Definitive evidence of safety and efficacy of intercostal nerve block and neurolysis for cancer pain will require a prospective randomized clinical trial.


PTOV1 expression predicts prostate cancer in men with isolated high-grade prostatic intraepithelial neoplasia in needle biopsy.
To analyze the expression of PTOV1 in high-grade prostatic intraepithelial neoplasia (HG-PIN) and to explore its usefulness to predict prostate cancer in patients with isolated HG-PIN in needle biopsy (prostate needle biopsy). PTOV1 expression in HG-PIN lesions from 140 patients was analyzed by immunohistochemistry in a semiquantitative manner (Histo-score). HG-PIN derived from 79 radical prostatectomies for prostate cancer and from 11 cistoprostatectomies for bladder cancer without prostate cancer were used as positive and negative controls, respectively. Fifty patients with HG-PIN without concomitant cancer at their first prostate needle biopsy were chosen as the study group. Patients were followed by a mean of 2.5 repeated prostate needle biopsies (1-5), during a mean period of 12.4 months (1-39). PTOV1 expression in HG-PIN from radical prostatectomies showed a significantly higher Histo-score (162.6) compared with specimens from cistoprostatectomies (67.0). In the study group, PTOV1 expression was significantly higher in samples with cancer in the follow-up (11 patients, 22%) compared with samples in which cancer was not detected (151.4 versus 94.6). PTOV1 expression was the only independent predictor of cancer in the multivariate analysis and the area under the curve was 0.803 (95% confidence interval, 0.728-0.878). A threshold of 100 for PTOV1 expression provided 90.9% sensitivity, 51.3% specificity, 34.5% positive predictive value, and 95.2% negative predictive value. PTOV1 is overexpressed in HG-PIN associated with cancer and is a potential marker for studying the carcinogenesis and progression of prostate cancer. Prostate needle biopsy with PTOV1 expression in HG-PIN above a threshold of 100 should be repeated immediately for the likely presence of undiagnosed cancer.


The regulation of exogenous NAD(P)H oxidation in spinach (Spinacia oleracea) leaf mitochondria by pH and cations.
Essentially chlorophyll-free mitochondria were isolated from green leaves of spinach (Spinacia oleracea L. cv. Viking II). Uncoupled oxidation of exogenous NADPH (1 mM) to oxygen had an optimum at pH 6.0, and activity was relatively low at pH 7.0, even in the presence of 1 mM-CaCl2. There was a proportional increase in the apparent Km for NADPH with decreasing H+ concentrations, suggesting that NADPH protonated on the 2'-phosphate group was the true substrate. Exogenous NADH was oxidized by oxygen with an optimum at pH 6.9. Under low-cation conditions, EGTA or EDTA (both 1 mM) had no effect on the Vmax. of NADH oxidation, although the removal of bivalent cations from the membrane surface by the chelators could be observed by use of 9-aminoacridine fluorescence. In contrast, under high-cation conditions, chelators lowered the Vmax. by about 50%, probably due to a better approach of the negatively charged chelators to the negative membrane surface than under low-cation conditions. In a low-cation medium, the Vmax. of NADH oxidation was increased by about 50% by the addition of cations. This was caused by a lowering of the size of the negative surface potential through charge screening. In contrast with other cations, La3+ inhibited NADH oxidation, possibly through binding to lipids essential for NADH oxidation. The apparent Km for NADH varied 6-fold in response to changes in the size of the surface potential, suggesting that the approach of the negatively charged NADH to the active site is hampered by the negative surface potential. The results demonstrate that the spinach leaf cell can regulate the mitochondrial NAD(P)H oxidation through several mechanisms: the pH; the cation concentration in general; and the concentration of Ca2+ in particular. The results also emphasize the importance of electrostatic considerations when investigating the kinetic behaviour of membrane-bound enzymes.


Concentration-Driven Assembly and Sol-Gel Transition of π-Conjugated Oligopeptides.
Advances in supramolecular assembly have enabled the design and synthesis of functional materials with well-defined structures across multiple length scales. Biopolymer-synthetic hybrid materials can assemble into supramolecular structures with a broad range of structural and functional diversity through precisely controlled noncovalent interactions between subunits. Despite recent progress, there is a need to understand the mechanisms underlying the assembly of biohybrid/synthetic molecular building blocks, which ultimately control the emergent properties of hierarchical assemblies. In this work, we study the concentration-driven self-assembly and gelation of π-conjugated synthetic oligopeptides containing different π-conjugated cores (quaterthiophene and perylene diimide) using a combination of particle tracking microrheology, confocal fluorescence microscopy, optical spectroscopy, and electron microscopy. Our results show that π-conjugated oligopeptides self-assemble into β-sheet-rich fiber-like structures at neutral pH, even in the absence of electrostatic screening of charged residues. A critical fiber formation concentration cfiber and a critical gel concentration cgel are determined for fiber-forming π-conjugated oligopeptides, and the linear viscoelastic moduli (storage modulus G' and loss modulus G″) are determined across a wide range of peptide concentrations. These results suggest that the underlying chemical structure of the synthetic π-conjugated cores greatly influences the self-assembly process, such that oligopeptides appended to π-conjugated cores with greater torsional flexibility tend to form more robust fibers upon increasing peptide concentration compared to oligopeptides with sterically constrained cores. Overall, our work focuses on the molecular assembly of π-conjugated oligopeptides driven by concentration, which is controlled by a combination of enthalpic and entropic interactions between oligopeptide subunits.


Mechanisms of selenium chemoprevention and therapy in prostate cancer.
Prostate cancer (PC) chemoprevention has generated considerable interest in the last decade and selenium and combinations of selenium have been recognized as one of the most efficacious chemopreventive agents against PC. This review focuses on a discussion of the knowledge hitherto gained about the mechanisms of action of the various in vitro and in vivo used selenium compounds and their effects on cellular processes and signaling pathways. We also describe the clinical and preclinical studies that have contributed enormously to the knowledge about dose, duration of exposure, and the chemical form of selenium effective in different scenarios. Even though the jury is still out about whether selenium can be used as a chemopreventive agent in the clinic and whether studies with cell lines and populations at low, medium, or high risk can adequately represent the physiological behavior of this micronutrient, it can safely be said to offer the most diverse spectrum of protective effects against this particular type of cancer which may augur well for its future as a chemopreventive agent.


The ototoxic effect of boric acid solutions applied into the middle ear of guinea pigs.
This study analyzed the ototoxic effects of boric acid solutions. Boric acid solutions have been used as otologic preparations for many years. Boric acid is commonly found in solutions prepared with alcohol or distilled water but can also be found in a powder form. These preparations are used for both their antiseptic and acidic qualities in external and middle ear infections. We investigated the ototoxic effect of boric acid solutions on guinea pigs. We are unaware of any similar, previously published study of this subject in English. The study was conducted on 28 young albino guinea pigs. Prior to application of the boric acid solution under general anesthesia, an Auditory Brainstem Response (ABRs) test was applied to the right ear of the guinea pigs. Following the test, a perforation was created on the tympanic membrane of the right ear of each guinea pig and small gelfoam pieces were inserted into the perforated area. Test solutions were administered to the middle ear for 10 days by means of a transcanal route. Fifteen days after inserting the gelfoams in all of the guinea pigs, we anasthesized the guinea pigs and removed the gelfoams from the perforated region of the ear and then performed an ABRs on each guinea pig. The ABRs were within the normal range before the applications. After the application, no significant changes were detected in the ABRs thresholds in neither the saline group nor the group administered boric acid and distilled water solution; however, significant changes were detected in the ABRs thresholds of the Gentamicine and boric acid and alcohol solution groups. We believe that a 4% boric acid solution prepared with distilled water can be a more reliable preparation than a 4% boric acid solution prepared with alcohol.


Aquatic Activity-Related Craniofacial Injuries Presenting to United States Emergency Departments, 2010 to 2019.
Aquatic activities are some of the most widely enjoyed sports and recreational activities in the United States. This study aimed to analyze the risks and types of craniofacial injuries associated with various aquatic activities. We retrospectively analyzed aquatic activity-related craniofacial injuries between 2010 and 2019 using the National Electronic Injury Surveillance System database. Aquatic activities included swimming, diving, surfing, water skiing, and water tubing. Risks and types of craniofacial injuries were analyzed according to the types of aquatic activities, age, and sex. Among 48,112 patients with aquatic activity-related injuries, 9,529 (19.8%) had craniofacial injuries. In decreasing order of frequency, the causes of craniofacial injuries were swimming (79.6%), diving (7.5%), surfing (5.9%), water skiing (3.6%), and water tubing (3.5%). The proportion of craniofacial injuries among all injuries was higher in males than in females (22.8 vs 16.3%, P < .001), and decreased with age: 27.7% in 0 to 5 years, 21.5% in 6 to 11 years, 20.5% in 12 to 17 years, and 15.2% in ≥18 years (P < .001). Additionally, the proportion of craniofacial injuries was highest in diving (38.7%), and lowest in swimming (18.1%, P < .001). The male-to-female ratio in the proportion of patients with craniofacial injuries was highest in swimming at 1.42, and close to unity (0.97-1.13) in other activities. The most common type of craniofacial injury was laceration (43.5%), followed by concussion/internal organ injury (38.9%), contusion/abrasion (11.3%), and fracture (3%). Among all craniofacial injuries, the proportion of craniofacial fractures was highest in water tubing (8.5%) and lowest in swimming (2.2%, P < .001). Craniofacial injuries are a frequent cause of morbidity related to aquatic activities. Substantial variability exists in the risk and pattern of craniofacial injuries depending on the type of activity, age, and sex. These findings may aid in instituting educational programs and preventive measures against aquatic activity-related craniofacial injuries.


Contribution of single genes within the unique short region of Aujeszky's disease virus (suid herpesvirus type 1) to virulence, pathogenesis and immunogenicity.
Pigs (3 and 10 weeks old) were infected intranasally with Aujeszky's disease virus (ADV) mutants that functionally lacked one of the non-essential genes in the unique short region of the genome (except the gene encoding the 11K protein). Virus excretion in oropharyngeal fluid and disease symptoms were monitored. Some pigs were killed to study pathogenesis, whereas others were challenged with virulent ADV 8 weeks after the primary infection. Mutants lacking protein kinase, or glycoproteins gp63 or gI showed reduced virulence, but mutants lacking gX or the 28K protein showed normal virulence. Glycoprotein gI appears to affect the tissue tropism of ADV in pigs, presumably by facilitating the spread of the virus through the central nervous system. In this study, there was no correlation between virulence and virus multiplication in either cultured cells or in the oropharynx in vivo. All mutants induced neutralizing antibody and complete or partial protection against challenge infection. Complete protection was obtained by inoculation with the gI and gX mutants, whereas incomplete protection was obtained using gp63 and protein kinase mutants. Complete clinical and virological protection was associated with the absence of secondary antibody responses in the serum.


Regulation of TG accumulation and lipid droplet morphology by the novel TLDP1 in Aurantiochytrium limacinum F26-b.
Thraustochytrids are marine single-cell protists that produce large amounts of PUFAs, such as DHA. They accumulate PUFAs in lipid droplets (LDs), mainly as constituent(s) of triacylglycerol (TG). We identified a novel protein in the LD fraction of Aurantiochytrium limacinum F26-b using 2D-difference gel electrophoresis. The protein clustered with orthologs of thraustochytrids; however, the cluster was evolutionally different from known PAT family proteins or plant LD protein; thus, we named it thraustochytrid-specific LD protein 1 (TLDP1). TLDP1 surrounded LDs when expressed as a GFP-tagged form. Disruption of the tldp1 gene decreased the content of TG and number of LDs per cell; however, irregular and unusually large LDs were generated in tldp1-deficient mutants. Although the level of TG synthesis was unchanged by the disruption of tldp1, the level of TG degradation was higher in tldp1-deficient mutants than in the WT. These phenotypic abnormalities in tldp1-deficient mutants were restored by the expression of tldp1 These results indicate that TLDP1 is a thraustochytrid-specific LD protein and regulates the TG accumulation and LD morphology in A. limacinum F26-b.


Genetic and hormonal regulation of esterase 6 activity in male Drosophila melanogaster.
The esterase 6 locus in Drosophila melanogaster is the structural gene for a carboxylesterase (E.C. 18.104.22.168) and is polymorphic for two major electrophoretic variants (slow and fast). Isogenic lines containing X chromosomes extracted from natural populations and substituted into a common genetic background were used to detect unlinked factors that affect the activity of the Est 6 locus. Twofold activity differences of esterase 6 were found among males from these derived lines, which differ only in their X chromosome. These unlinked activity modifiers identify possible regulatory elements. Immunoelectrophoresis was used to estimate quantitatively the levels of specific cross-reacting material in the derived lines. The results show that the variation in activity is due to differences in the amount of EST 6 present. Physiological studies of the control of EST 6 levels reveal that both juvenile hormone and 20 hydroxyecdysone stimulate the production of EST 6 activity in adult males. These results suggest that the effects of X chromosomes on EST 6 activity may be effected by modulating the level of adult hormone titers in Drosophila.


Phenol sulphotransferase SULT1A1 polymorphism: molecular diagnosis and allele frequencies in Caucasian and African populations.
Sulphation, catalysed by members of the sulphotransferase (SULT) enzyme family, is an important component of the body's chemical defence mechanism, but also acts to bioactivate mutagens such as hydroxylated aryl and heterocyclic amines. A major human sulphotransferase, SULT1A1 (P-PST), metabolizes and/or bioactivates many drugs, iodothyronines and hydroxylated aromatic amines. The enzyme activity varies widely within the population and is under genetic control. We have developed an assay detecting a G-->A transition in SULT1A1 that causes an Arg213-->His substitution associated with low SULT activity and altered enzyme properties, and have used it to assess the SULT1A1 genotype in Caucasian (n=293) and African (Nigerian, n=52) populations. We show that the mutant SULT1A1*2 allele is present at frequencies of 0.321 and 0.269 in the Caucasian and African populations respectively. We also demonstrate a significant age-related difference in SULT1A1 genotype within our Caucasian population, with increasing incidence of SULT1A1*1 homozygosity and decreasing incidence of SULT1A1*2 homozygosity with increasing age, indicating a potential association of SULT1A1*1 allozyme(s) with protection against cell and/or tissue damage during aging.


A systematic review and meta-analysis of pregnancy outcomes in patients with systemic lupus erythematosus and lupus nephritis.
Studies of the impact of systemic lupus erythematosus (SLE) and its pregnancy complications have yielded conflicting results. Major limitations of these studies relate to their small numbers of patients and retrospective designs. The aim of this study was to perform a systematic literature review of pregnancy outcomes in women with SLE and a meta-analysis of the association of lupus nephritis with adverse pregnancy outcomes. We searched electronic databases from 1980 to 2009 and reviewed papers with validity criteria. Random-effects analytical methods were used to evaluate pregnancy complications rates. Thirty-seven studies with 1842 patients and 2751 pregnancies were included. Maternal complications included lupus flare (25.6%), hypertension (16.3%), nephritis (16.1%), pre-eclampsia (7.6%), and eclampsia (0.8%). The induced abortion rate was 5.9%, and when excluded, fetal complications included spontaneous abortion (16.0%), stillbirth (3.6%), neonatal deaths (2.5%), and intrauterine growth retardation (12.7%). The unsuccessful pregnancy rate was 23.4%, and the premature birth rate was 39.4%. Meta-regression analysis showed statistically significant positive associations between premature birth rate and active nephritis and increased hypertension rates in subjects with active nephritis or a history of nephritis. History of nephritis was also associated with pre-eclampsia. Anti-phospholipid antibodies were associated with hypertension, premature birth, and an increased rate of induced abortion. In patients with SLE, both lupus nephritis and anti-phospholipid antibodies increase the risks for maternal hypertension and premature births. The presented evidence further supports timing of pregnancy relative to SLE activity and multispecialty care of these patients.


Estimating the lost benefits of antiretroviral drug use in South Africa.
South Africa is one of the countries most severely affected by HIV/AIDS. At the peak of the epidemic, the government, going against consensus scientific opinion, argued that HIV was not the cause of AIDS and that antiretroviral (ARV) drugs were not useful for patients and declined to accept freely donated nevirapine and grants from the Global Fund. Using modeling, we compared the number of persons who received ARVs for treatment and prevention of mother-to-child HIV transmission between 2000 and 2005 with an alternative of what was reasonably feasible in the country during that period. More than 330,000 lives or approximately 2.2 million person-years were lost because a feasible and timely ARV treatment program was not implemented in South Africa. Thirty-five thousand babies were born with HIV resulting in 1.6 million person-years lost by not implementing a mother-to-child transmission prophylaxis program using nevirapine. The total lost benefits of ARVs are at least 3.8 million person-years for the period 2000-2005.


Seroprevalence and Molecular Detection of Brucellosis in Hospitalized Patients in Lahore Hospitals, Pakistan.
Brucellosis is one of the most notorious zoonoses worldwide. The disease is common and endemic in humans and animals of Pakistan, but lack of awareness and lack of research have resulted in an increased incidence in the human population. The present study aimed to determine the seroprevalence and at molecular detection of brucellosis in patients with clinical symptoms in six different hospitals from Lahore, which is the capital city of Punjab province. A total of 218 blood samples were collected from hospitalized patients. The samples were initially screened by the Rose Bengal Plate Test (RBPT), and then quantitative real-time PCR (qRT-PCR) was applied. An overall seroprevalence of 17% (37/218) was found. The highest prevalence was found at the Lady Health center (36.53%), which was followed by the Lady Willingdon Hospital (28.6%). Female patients showed a higher seroprevalence than males and peaked at 34% (n = 32) for women who suffered from abortion. In total, 16.8% of patients younger than 30 years showed seropositive reactions, while the prevalence was 19% in patients between 31 and 50. Thirty-three DNA samples from 24 seropositive and nine seronegative patients tested positive, 32 samples were found positive for B. abortus DNA, and one sample failed to be identified at the species level. Almost all positive cases had direct contact with animals and consumed unpasteurized dairy products. Research on human brucellosis is still scarce in Pakistan. For the diagnosis of brucellosis, serology and molecular tools should be combined if isolation by culture is not possible. Nationwide control activities and increasing awareness for zoonotic brucellosis are needed.


Transforming growth factor-beta profiles correlate with clinical symptoms and parameters of haemostasis and inflammation in a controlled human malaria infection.
After a controlled human malaria infection (CHMI), presentation of clinical signs and symptoms and host responses is heterogeneous. Transforming growth factor-beta (TGF-β) is the first serum cytokine that changes in malaria-naïve volunteers after CHMI. We studied a possible relation between TGF-β changes, pro-inflammatory cytokines, activation of haemostasis and endothelial cells and clinical symptoms. A panel of cytokines including TGF-β, and markers of activation of haemostasis and endothelial cells were measured in blood samples of 15 volunteers at baseline before CHMI and during CHMI at day of treatment. The change of the parameters on the day of treatment was examined for a significant alteration during infection. Nine of 15 volunteers showed a significant decrease in TGF-β compared to baseline, with concomitant increased concentrations of D-dimer (p = 0.012), Von Willebrand factor (p = 0.017), IL-6 (p = 0.012) and IFN-γ (0.028) and a significantly decreased platelet count (p = 0.011). In contrast, 6 of 15 volunteers showed sustained or increased TGF-β concentrations without change in the aforementioned parameters. The sustained responders presented with less moderate and severe clinical symptoms than the negative responders (p = 0.036) and had a higher baseline lymphocyte count (p = 0.026). TGF-β concentrations did not correlate with the parasitaemia on day of treatment. Early decreases of serum TGF-β might function a marker for a pro-inflammatory host response and downstream clinical symptoms and pathology during CHMI.


[Biochemical indices of the allantoic fluid of chick embryos infected with the influenza virus].
Virus-containing allantoic fluid (VCAF) serves as raw material for influenza virus manufacture in the USSR. VCAF composition varies greatly from batch to batch. The causes of instability of its composition were studied. The main high molecular admixtures determining VCAF instability were found to include proteins and lipids getting into VCAF with amniotic fluid, blood, and yolk of the embryo when VCAF is collected. Physicochemical characteristics (pH, protein and lipid concentrations, protein composition) for VCAF control at pooling are proposed.


Vaccine strategies for prevention of community-acquired pneumonia in Canada: Who would benefit most from pneumococcal immunization?
To describe the burden of pneumococcal disease and associated risk factors in the Canadian adult population, delineate available pneumococcal vaccines and associated efficacy and effectiveness data, and review current pneumococcal vaccine recommendations and community-acquired pneumonia (CAP) prevention strategies in Canada. Pneumococcal vaccination guidelines from the Canadian National Advisory Committee on Immunization in 2013 and 2016 constitute level III evidence for CAP prevention in the Canadian adult population. It is recommended that immunosuppressed adults of all ages receive the 13-valent pneumococcal conjugate vaccine (PCV13) (grades A and B recommendations). In 2016, the National Advisory Committee on Immunization also recommended that all adults aged 65 years and older receive PCV13 (grade A recommendation) on an individual basis, followed by the 23-valent pneumococcal polysaccharide vaccine (grade B recommendation). This update is based on a large clinical study that demonstrated PCV13 efficacy against vaccine-type CAP in this population. Physicians should focus on improving pneumococcal vaccination rates among adults, which remain low. Vaccination with PCV13 should also be considered for adults with chronic conditions, whose baseline risk is often higher than that for healthy individuals aged 65 years and older.


Interaction of Epididymal Epithelia and their Secretions with Spermatozoa Supports Functional and Morphological Changes During Long-Term Storage in the Chinese Soft-Shelled Turtle (Pelodiscus sinensis).
Post-testicular maturation of spermatozoa is crucial for attaining the morphological and functional capabilities needed for successful fertilization. Epididymal epithelia offer a favorable environment for spermatozoa that are stored long term in the turtle epididymis; however, sperm-epithelial interactions during storage, which are enormously important for sperm functional and morphological maturation, are still largely unknown in turtles. The present study examined the epididymis during the sperm-storage period (November-April) in the Chinese soft-shelled turtle (Pelodiscus sinensis). Light and transmission electron microscopy were used to determine the cellular features of each epididymal segment (caput, corpus, and cauda) and their epithelial interactions with the spermatozoa. Spermatozoa were mainly located in the lumena of caput, corpus, and cauda epididymides. Numerous spermatozoa were bound to apical surfaces of the epithelia, and several were even embedded in the epithelial cytoplasm of the caput and corpus epididymides. No embedded spermatozoa were found in the cauda epididymis. In all epididymal segments, principal and clear cells showed the synthetic activity, evidenced by a well-developed endoplasmic reticulum network and high and low electron-dense secretory materials, respectively. Principal and clear cells in the caput and corpus segments showed embedded spermatozoa in electron-dense secretions and in the lipid droplets within the cytoplasm. No lysosomes were observed around the embedded spermatozoa. The lumena of the caput and corpus segments showed few apocrine and low electron density secretions. In the lumen of the cauda epididymidis, different secretions, such as holocrine with low and high electron density and their fragmentation, apocrine, and dictyosome, were found and are summarized. Altogether, sperm physical interactions with secretions either in the cytoplasm of epithelium or in the lumen may support the viability, morphological maintenance, and transfer of various proteins involved in long-term sperm storage in the turtle. This interaction could help us to understand the mechanisms of long-term sperm storage and provide more insights into the reproductive strategies of turtle sperm preservation.


[Neuroleptics in psychogeriatrics. A proposal for a more rational prescription policy].
All neuroleptics have a cerebral dopamine receptor blocking effect. This is the best documented explanation for their anti-psychotic effect. Furthermore, they act to a widely differing extent in a blocking manner to other central neurotransmitters, such as noradrenaline, acetylcholine, histamine, and serotonin. During the last couple of years quantitative data have been produced, enabling a more rational choice of neuroleptics. Based on these data and a number of pharmacokinetic considerations, a provisional scheme has been elaborated for the choice of neuroleptics in some clinical situations in psychogeriatrics. Our own experiences with this scheme are promising. It allowed us to restrict the use of neuroleptics to five drugs. The neuroleptics with a strong sedative and anticholinergic action, such as alimemazine, promazine and levomepromazine, being potentially dangerous to particularly elderly patients, are no longer used.


ADP ribose polymerase inhibitors for treating non-small cell lung cancer: new additions to the pharmacotherapeutic armamentarium.
Poly (ADP-ribose) polymerase inhibitors (PARPi) are already part of the armamentarium of drugs available against ovarian and breast cancer. There is less data available on the efficacy of these drugs in the treatment of non-small cell lung cancer (NSCLC). The authors have analyzed the preclinical studies that justified the use of PARPi in NSCLC. They then evaluate the in vivo efficacy of the combination of these drugs with chemotherapy, radiotherapy, and immunotherapy. Data from clinical trials available to date have discouraged the use of PARPi in association with chemotherapy or radiotherapy in NSCLC. The knowledge available to date opens the door to the use of PARPi in association with immunotherapy. In fact, the activity of these drugs would not be based only on direct cytotoxic action, but also on the modification of the intra-tumor microenvironment, in particular by increasing the expression of PD-L1 on tumor cells. This action might potentially enhance available treatments with a modest increase in toxicity.


Digital dermatitis: report of an outbreak.
An outbreak of digital dermatitis in a dairy herd in the Gauteng province of South Africa is described in which 72% of the lactating herd was affected. Many of the affected cows showed a severe lameness and a drop in milk production. Other complications included heel horn erosion, under-running of heel horn and boxy claws. The diagnosis was made on the typical clinical appearance of the condition and the presence of spirochaetes on histopathological sections from biopsy material. The condition responded to spraying affected feet with an oxytetracycline mixture after cleaning with water using a high pressure hose. The prevalence was reduced to 28% after one month of therapy. A repeat outbreak occurred on the same farm 7 months later during which time 37% of the lactating herd was affected. Of the cows affected, 48% represented new cases, whereas the rest were reinfections.


Increased Incidence of juvenile onset systemic lupus erythematosus in children with atopic dermatitis.
Herein, we investigated the risk of juvenile-onset systemic lupus erythematosus (JSLE) in children with atopic dermatitis (AD). From 2000 to 2007, 192,357 children with newly diagnosed AD and 769,428 matched non-AD controls were identified. By the end of 2008, incidences and hazard ratios (HRs) of JSLE were measured. JSLE incidence in the AD cohort was 2.90-fold greater than that in the non-AD cohort (3.25 vs. 1.12 per 100,000 person-years), with a Cox model-measured adjusted HR of 2.92 (95% CI: 1.85-4.60); the risk of JSLE was greater for older children and girls. The AD-to-non-AD cohort HR was 6.6 (95% CI: 2.88-13.1) for children aged >12 years compared with 1.81 (95% CI: 0.98-3.32) for children aged ≤ 12 years. The HR of JSLE in AD children increased from 1.55 (95% CI: 0.88-2.76) for those with ≤ 3 clinical visits to 66.3 (95% CI: 33.1-132.8) for those with >6 visits (p < 0.0001, by trend test). The risk of developing SLE in the AD cohort was the highest within five years after AD diagnosis (HR: 4.02; 95% CI: 2.83-7.08). Children with AD are at a high risk of developing JSLE during their growth period.


Use of the cuffed oropharyngeal airway for manual ventilation by nonanaesthetists.
We studied the use of the cuffed oropharyngeal airway in 100 ASA I and II anaesthetised patients. In the first 50 patients (group A), an experienced anaesthetist inserted the airway. The optimum sizes and cuff volumes for manual ventilation in adult males and females were found to be sizes 11 and 10 with up to 60 ml and 50 ml in each cuff, respectively. Manual ventilation was clinically successful in 49/50 (98%) of these patients. Using these recommendations and following a brief tutorial, a group of 50 nonanaesthetic, basic life-support providers attempted to insert a cuffed oropharyngeal airway and manually ventilate the lungs of a subsequent 50 patients (group NA). Clinically adequate tidal volumes were achieved within 45 s in 47/50 (94%) patients in this group. A persistent leak was present in 21/49 (43%) and 24/47 (51%) of the successful insertions in each group, but this did not affect the ability to ventilate the lungs adequately. The cuffed oropharyngeal airway may offer an effective method of providing adequate ventilation during resuscitation by nonanaesthetic hospital staff.


The impact of formulation attributes and process parameters on black seed oil loaded liposomes and their performance in animal models of analgesia.
This study aimed to formulate black seed oil (Nigella sativa) loaded liposomes using the ethanol injection method to enhance oral bioavailability and improve therapeutic activity in small animal studies of analgesia. The impact of formulation attributes and process parameters on the liposomal system was evaluated with key quality attributes being particle size, morphology, and entrapment efficiency. The particle size and entrapment efficiency of the liposome preparation were found to be between the range of 50-900 nm and 34-87% respectively. Particle size distribution data suggested that increasing the percentage of oil, up to a certain concentration, reduced the size of the liposomes significantly from 520 ± 81.2 nm to 51.48 ± 1.31 nm. Stirring and injection rate were shown to have marked impact on the average particle size of liposome. It was observed that entrapment efficiency of liposomes was greatly influenced by the amount of cholesterol and type of cryoprotectant used during formulation. The stability study indicated that the liposomal preparation was stable at ambient conditions for one month. In vivo studies showed that the liposomal preparation demonstrated significant analgesic activity in mice.


Aquaporin 11 variant associates with kidney disease in type 2 diabetic patients.
Kidney disease, a common complication of diabetes, associates with poor prognosis. Our previous animal model studies linked aquaporin (AQP)11 to acute kidney injury, hyperglycemia-induced renal impairment, and kidney disease in diabetes. Here, we report the AQP11 rs2276415 variant as a genetic factor placing type 2 diabetic patients at greater risk for the development of kidney disease. We performed two independent retrospective case-control studies in 1,075 diabetic and 1,619 nondiabetic individuals who were identified in the Synthetic Derivative Database with DNA samples in the BioVU DNA repository at Vanderbilt University (Nashville, TN). A χ(2)-test and multivariable logistic regression analysis with adjustments for age, sex, baseline serum creatinine, and underlying comorbid disease covariates showed a significant association between rs2276415 and the prevalence of any event of acute kidney injury and chronic kidney disease (CKD) in diabetic patients but not in patients without diabetes. This result was replicated in the second independent study. Diabetic CKD patients over 55 yrs old with the minor AQP11 allele had a significantly faster progression of estimated glomerular filtration rate decline than patients with the wild-type genotype. Three-dimensional structural analysis suggested a functional impairment of AQP11 with rs2276415, which could place diabetic patients at a higher risk for kidney disease. These studies identified rs2276415 as a candidate genetic factor predisposing patients with type 2 diabetes to CKD.


Beneficial effects of Apelin-13 on metabolic diseases and exercise.
Apelin, a novel endogenous ligand of the G-protein-coupled receptor APJ, is encoded by the APLN gene and can be hydrolyzed into multiple subtypes, with Apelin-13 being one of the most active subtypes of the Apelin family. Recent studies have revealed that Apelin-13 functions as an adipokine that participates in the regulation of different biological processes, such as oxidative stress, inflammation, apoptosis, and energy metabolism, thereby playing an important role in the prevention and treatment of various metabolic diseases. However, the results of recent studies on the association between Apelin-13 and various metabolic states remain controversial. Furthermore, Apelin-13 is regulated or influenced by various forms of exercise and could therefore be categorized as a new type of exercise-sensitive factor that attenuates metabolic diseases. Thus, in this review, our purpose was to focus on the relationship between Apelin-13 and related metabolic diseases and the regulation of response movements, with particular reference to the establishment of a theoretical basis for improving and treating metabolic diseases.


Studying the complex expression dependences between sets of coexpressed genes.
Organisms simplify the orchestration of gene expression by coregulating genes whose products function together in the cell. The use of clustering methods to obtain sets of coexpressed genes from expression arrays is very common; nevertheless there are no appropriate tools to study the expression networks among these sets of coexpressed genes. The aim of the developed tools is to allow studying the complex expression dependences that exist between sets of coexpressed genes. For this purpose, we start detecting the nonlinear expression relationships between pairs of genes, plus the coexpressed genes. Next, we form networks among sets of coexpressed genes that maintain nonlinear expression dependences between all of them. The expression relationship between the sets of coexpressed genes is defined by the expression relationship between the skeletons of these sets, where this skeleton represents the coexpressed genes with a well-defined nonlinear expression relationship with the skeleton of the other sets. As a result, we can study the nonlinear expression relationships between a target gene and other sets of coexpressed genes, or start the study from the skeleton of the sets, to study the complex relationships of activation and deactivation between the sets of coexpressed genes that carry out the different cellular processes present in the expression experiments.


Diet and inflammation.
The emerging role of chronic inflammation in the major degenerative diseases of modern society has stimulated research into the influence of nutrition and dietary patterns on inflammatory indices. Most human studies have correlated analyses of habitual dietary intake as determined by a food frequency questionnaire or 24-hour recall with systemic markers of inflammation like high-sensitivity C-reactive protein (HS-CRP), interleukin-6 (IL-6), and tumor necrosis factor alpha (TNF-α). An occasional study also includes nutrition analysis of blood components. There have been several controlled interventions which evaluated the effect of a change in dietary pattern or of single foods on inflammatory markers in defined populations. Most studies reveal a modest effect of dietary composition on some inflammatory markers in free-living adults, although different markers do not vary in unison. Significant dietary influences have been established for glycemic index (GI) and load (GL), fiber, fatty acid composition, magnesium, carotenoids, and flavonoids. A traditional Mediterranean dietary pattern, which typically has a high ratio of monounsaturated (MUFA) to saturated (SFA) fats and ω-3 to ω-6 polyunsaturated fatty acid (PUFAs) and supplies an abundance of fruits, vegetables, legumes, and grains, has shown anti-inflammatory effects when compared with typical North American and Northern European dietary patterns in most observational and interventional studies and may become the diet of choice for diminishing chronic inflammation in clinical practice.


Cryoablation of unresectable pancreatic cancer.
Most patients with pancreatic cancer are not candidates for curative resection. The goal of this study was to evaluate the safety of an intraoperative ultrasound-guided cryosurgical procedure in a phase I study of unresectable pancreatic cancer. From March 1995 to March 1999, 10 cryosurgeries using intraoperative ultrasound were performed on 9 patients with unresectable cancers at laparotomy. Four patients had a concurrent gastrojejunostomy, 2 had a chemical splanchnicectomy, and 1 underwent a concurrent hepatic cryosurgical procedure. There was no intraoperative morbidity or mortality. No patients developed postoperative pancreatitis or fistula. All patients had good pain control postoperatively and were tolerating a regular diet at the time of discharge. Pain control at discharge was achieved with an oral formulation (4/9), transdermal patch (3/9), no pain medication (1/9), and intravenous patient controlled analgesia (1/9). Ultrasound-guided cryoablation for unresectable pancreatic cancer appears safe and may contribute to improved postoperative pain control. Future studies to determine its therapeutic role in the management of unresectable pancreatic cancer are indicated.


Modulation of active Ca2+ uptake by the islet-cell endoplasmic reticulum.
The possible effects of calmodulin and cyclic AMP on active Ca2+ uptake by the islet-cell endoplasmic reticulum were investigated. Neither calmodulin nor cyclic AMP affected the rate of active Ca2+ uptake, or the steady-state filling capacity of the endoplasmic reticulum when measured in the absence of oxalate. Consistent with these results, calmodulin did not activate the Ca2+-stimulated ATPase activity associated with this cell fraction. During the course of these experiments., it was unexpectedly discovered that the rate of Ca2+ uptake, as well as the steady-state Ca2+ filling capacity of the endoplasmic reticulum, were markedly increased by unidentified factor(s) in the cytosol. This effect could be demonstrated by reconstitution of the membranes in cytosol, or by direct addition of fresh or dialysed cytosol to the Ca2+ uptake assays. The degree of activation by the cytosol indicates that the endoplasmic reticulum may play a prominent role in controlling beta-cell Ca2+ concentrations and that the unidentified activator(s) present in the cytosol may be involved in regulation of this function.


Plaque formation by Chlamydia in L cells.
Chlamydiae were found capable of producing plaques in several cell lines. Mouse fibroblast cells, L-929, proved the most sensitive to infection and yielded plaques of the highest clarity. Assay of chlamydial infectivity by plaque titration was at least as sensitive as egg ld(50) determination. Among chlamydial isolates of avian, mammalian, and human origin, only slow-growing trachoma-inclusion-conjunctivitis agents did not produce plaques. The plaque assay is highly sensitive, reproducible, and offers a potential tool for investigations requiring accurate measurement of small changes in chlamydial infectivity.


Antibiotic susceptibility of coagulase-negative staphylococci isolated from very low birth weight babies: comprehensive comparisons of bacteria at different stages of biofilm formation.
Coagulase-negative staphylococci are major causes of bloodstream infections in very low birth weight babies cared for in Neonatal Intensive Care Units. The virulence of these bacteria is mainly due to their ability to form biofilms on indwelling medical devices. Biofilm-related infections often fail to respond to antibiotic chemotherapy guided by conventional antibiotic susceptibility tests. Coagulase-negative staphylococcal blood culture isolates were grown in different phases relevant to biofilm formation: planktonic cells at mid-log phase, planktonic cells at stationary phase, adherent monolayers and mature biofilms and their susceptibilities to conventional antibiotics were assessed. The effects of oxacillin, gentamicin, and vancomycin on preformed biofilms, at the highest achievable serum concentrations were examined. Epifluorescence microscopy and confocal laser scanning microscopy in combination with bacterial viability staining and polysaccharide staining were used to confirm the stimulatory effects of antibiotics on biofilms. Most coagulase-negative staphylococcal clinical isolates were resistant to penicillin G (100%), gentamicin (83.3%) and oxacillin (91.7%) and susceptible to vancomycin (100%), ciprofloxacin (100%), and rifampicin (79.2%). Bacteria grown as adherent monolayers showed similar susceptibilities to their planktonic counterparts at mid-log phase. Isolates in a biofilm growth mode were more resistant to antibiotics than both planktonic cultures at mid-log phase and adherent monolayers; however they were equally resistant or less resistant than planktonic cells at stationary phase. Moreover, for some cell-wall active antibiotics, concentrations higher than conventional MICs were required to prevent the establishment of planktonic cultures from biofilms. Finally, the biofilm-growth of two S. capitis isolates could be enhanced by oxacillin at the highest achievable serum concentration. We conclude that the resistance of coagulase-negative staphylococci to multiple antibiotics initially remain similar when the bacteria shift from a planktonic growth mode into an early attached mode, then increase significantly as the adherent mode further develops. Furthermore, preformed biofilms of some CoNS are enhanced by oxacillin in a dose-dependent manner.


Effect of Divalent Cations on RED Performance and Cation Exchange Membrane Selection to Enhance Power Densities.
Reverse electrodialysis (RED) is a membrane-based renewable energy technology that can harvest energy from salinity gradients. The anticipated feed streams are natural river and seawater, both of which contain not only monovalent ions but also divalent ions. However, RED using feed streams containing divalent ions experiences lower power densities because of both uphill transport and increased membrane resistance. In this study, we investigate the effects of divalent cations (Mg2+ and Ca2+) on RED and demonstrate the mitigation of those effects using both novel and existing commercial cation exchange membranes (CEMs). Monovalent-selective Neosepta CMS is known to block divalent cations transport and can therefore mitigate reductions in stack voltage. The new multivalent-permeable Fuji T1 is able to transport divalent cations without a major increase in resistance. Both strategies significantly improve power densities compared to standard-grade CEMs when performing RED using streams containing divalent cations.


Urinary tract injury associated with pelvic fractures.
Pelvic trauma associated with urinary tract injury is a severe trauma, mostly caused by traffic accidents and falls from heights. These injuries require urgent treatment and close teamwork between urologic and orthopaedic surgeons. In this retrospective study there were analyzed patients with pelvic trauma and extraperitoneal injury of urinary tract, treated surgically at Clinic for Ortopaedic Surgery and Traumatology and Urology Clinic in Clinical Center Nis. Surgical intervention in these patients had been realized as the synchronized work of both orthopaedic and urologic surgeons. The pelvis was treated by external and internal fixation. Mitkovic type external fixator was used for pelvic external fixation. Plating was used for pelvic internal fixation. Pelvic fractures were classified using Tile's classification system. The final functional results had been scored using Majeed score system. There were 42 patients with the injury of pelvic ring, treated at Clinic for Ortopaedic Surgery and Traumatology and at Urology Clinic, Clinical Center Nis, in the period of 01.01.2011. to 31.12.2013, 30 males and 12 females, with average age of 53.69 (19-84) years old. In 80% of cases pelvic fractures were caused by high energy trauma in traffic accidents. According to Tile's classification, 9 patients (21,42%) had pelvic fracture type A, 23 patients (54,46%) had pelvic fracture type B and 10 patients (23,80%) had pelvic fracture type C. Urinary tract injury was diagnosed in 9 patients (21,42%): 5 patients (11,9%) with bladder injury, 3 patients (7,14%) with posterior urethra injury and 1 patient (2,38%) with both bladder and posterior urethra injury. Urgent repair of extraperitoneal urinary tract injury by urologic surgeons and synchronized pelvic reduction and fixation using external or internal fixation by ortopaedic surgeon, in the same surgical procedure, is the standard method for treatment of this severe injury.


Subclinical hypothyroidism and the risk of hypercholesterolemia.
Subclinical hypothyroidism (SCH) is a condition wherein there are small elevations in thyroid stimulating hormone (TSH), yet normal circulating levels of free thyroid hormones (FT4). Alterations in thyroid function result in changes in composition and transport of lipoproteins. The study was undertaken in the outpatient department of Lourdes hospital, Ernakulam. 110 patients between 40-69 years were studied, of which 60 were identified to have SCH based on cross-sectional survey, 50 patients were randomly selected to represent the EU group. Data based on interview, clinical examination, thyroid function, lipid profiles. Student's t, chi-square tests used for computation of p values. SCH were seen in females (66.66%) and in the age group of 40-49 years (61.66%). Mean cholesterol values were elevated in the subclinical hypothyroid group and in relation to age (60-69) and gender. Statistical analysis showed significant difference in total cholesterol(TC) (P<0.005) and triglycerides (P<0.05) in relation to age (40-49 yrs.) between two groups. Based on TSH, group A 5-7.49Mu/L and group B 7.5-10Mu/L were compared to the euthyroid group N<5Mu/L. Mean cholesterol values were raised in both subgroups. Statistically significant difference seen (P<0.0005) in cholesterol values between the subclinical hypothyroid group B and the euthyroid group N. SCH appears to be associated with increased mean cholesterol levels in females and of age > 60 years. The TC values were elevated in both subgroups of patients with SCH (A and B) based on TSH values.


Measuring functional and quality of life outcomes following major head injury: common scales and checklists.
Traumatic brain injury (TBI) is a major public health issue, which results in significant mortality and long term disability. The profound impact of TBI is not only felt by the individuals who suffer the injury but also their care-givers and society as a whole. Clinicians and researchers require reliable and valid measures of long term outcome not only to truly quantify the burden of TBI and the scale of functional impairment in survivors, but also to allow early appropriate allocation of rehabilitation supports. In addition, clinical trials which aim to improve outcomes in this devastating condition require high quality measures to accurately assess the impact of the interventions being studied. In this article, we review the properties of an ideal measure of outcome in the TBI population. Then, we describe the key components and performance of the measurement tools most commonly used to quantify outcome in clinical studies in TBI. These measurement tools include: the Glasgow Outcome Scale (GOS) and extended Glasgow Outcome Scale (GOSe); Disability Rating Scale (DRS); Functional Independence Measure (FIM); Functional Assessment Measure (FAM); Functional Status Examination (FSE) and the TBI-specific and generic quality of life measures used in TBI patients (SF-36 and SF-12, WHOQOL-BREF, SIP, EQ-5D, EBIQ, and QOLIBRI).


Rapid microwave-stimulated fixation of entire prostatectomy specimens. Biomed-II MPC Study Group.
Conventional fixation of large solid surgical specimens is a slow process. Consequently, autolytic damage to tissues may occur if the fixative does not reach the central part of the specimen in time. However, as there is also a time relationship between formalin fixation and antigen masking, fixation for too long can also be detrimental. In seeking the optimum balance for fixation, microwave irradiation might be of assistance. This study set out to evaluate methods for fixing entire prostate glands within a brief period of time, using microwave-stimulated formalin fixation. The results show that entire prostates can be optimally fixed if formalin is present throughout the tissue as the temperature is increased by microwave irradiation. This is achieved by injecting the fixative into the prostate at multiple sites immediately following prostatectomy. The technique described ensures standardization of a critical step during tissue processing, leading to uniform microscopic results with both routine and immunohistochemical stains. It is a simple, rapid method, suitable for routine diagnostic use. Using this modified approach, DNA of much larger sizes can be extracted from paraffin-embedded material, which could expand the possibilities for molecular analysis.


Redox modulatory site of the NMDA receptor-channel complex: regulation by oxidized glutathione.
We monitored increases in both intracellular calcium concentration [( Ca2+]i) and whole-cell current responses induced by N-methyl-D-aspartate (NMDA), applied with co-agonist glycine, using fura-2 digital imaging and patch-clamp recording techniques. Extracellular application of oxidized glutathione (GSSG), but not reduced glutathione (GSH), inhibited responses mediated by activation of the NMDA subtype of glutamate receptor in cultures of rat cortical and retinal ganglion cell neurons. The NMDA responses were persistently inhibited by GSSG (500 microM to 10 mM) until introduction of a selective sulfhydryl reducing agent, dithiothreitol, which resulted in complete recovery of the responses. Exposure of the neurons to 5,5-dithio-bis-2-nitrobenzoic acid (DTNB), an efficacious oxidizing agent, also resulted in persistently smaller responses to NMDA. The addition of GSSG following exposure to DTNB, however, did not result in a further decrement in NMDA responses in our experimental paradigm. These findings suggest that a predominant action of GSSG is oxidation of vicinal thiol groups to form a peptide disulfide bond(s) comprising the redox modulatory site of the NMDA receptor-channel complex. Evidence for such regulatory sulfhydryl centers associated with the NMDA receptor has been presented previously. Moreover, the fact that DTNB produced little if any additional attenuation of the NMDA [Ca2+]i response when administered after GSSG implies that GSSG is also an efficacious oxidant at this site. GSSG displayed little or no effect on [Ca2+]i responses elicited by high extracellular K+ or by kainate, suggesting that, at least under the conditions of the present experiments, GSSG was somewhat selective for the NMDA redox modulatory site. Based on these observations, we suggest that GSSG exerts its NMDA-specific redox effects in a novel extracellular manner.


Belimumab therapy for refractory immune thrombocytopenia in systemic lupus erythematosus patients with anti-phospholipid antibodies.
To observe the effect of belimumab treatment in refractory anti-phospholipid antibody-associated immune thrombocytopenia with systemic lupus erythematosus (SLE). Four SLE patients with refractory anti-phospholipid antibody-associated immune thrombocytopenia were included in this one-arm observational study. Inclusion criteria were a diagnosis of SLE according to 1997 American College of Rheumatology criteria, severe immune thrombocytopenia (platelets <30 × 109/L), no bleeding symptoms, lack of satisfactory response to traditional treatment, and high-titre anti-phospholipid antibodies. All patients received belimumab (Benlysta®) for 6 months. The mean platelet count was 21.8 × 109 cells/L, ranging between 16 and 29 × 109/L at baseline, 123.3 × 109/L at 1 month, and 172.5 × 109/L at the end of 6 months after belimumab treatment. No bleeding complications occurred during the entire follow-up period. In this study, belimumab reduced the anti-phospholipid antibodies while increasing the platelet count in SLE patients with anti-phospholipid antibody-associated immune thrombocytopenia.


Peripheral muscle ergoreceptors and ventilatory response during exercise recovery in heart failure.
Recent studies have suggested that the increased ventilatory response during exercise in patients with chronic heart failure was related to the activation of muscle metaboreceptors. To address this issue, 23 patients with heart failure and 7 normal subjects performed arm and leg bicycle exercises with and without cuff inflation around the arms or the thighs during recovery. Obstruction slightly reduced ventilation and gas exchange variables at recovery but did not change the kinetics of recovery of these parameters compared with nonobstructed recovery: half-time of ventilation recovery was 175 +/- 54 to 176 +/- 40 s in patients and 155 +/- 66 to 127 +/- 13 s in controls (P < 0.05, patients vs. controls, not significant within each group from baseline to obstructed recovery). We conclude that muscle metaboreceptor activation does not seem to play a role in the exertion hyperventilation of patients with heart failure.


Structure-activity relationships of the melanocortin tetrapeptide Ac-His-DPhe-Arg-Trp-NH(2) at the mouse melanocortin receptors: part 2 modifications at the Phe position.
The melanocortin pathway is an important participant in skin pigmentation, steroidogenesis, obesity, energy homeostasis and exocrine gland function. The centrally located melanocortin-3 and melanocortin-4 receptors (MC3R, MC4R) are involved in the metabolic and food intake aspects of energy homeostasis and are stimulated by melanocortin agonists such as alpha-melanocyte stimulation hormone (alpha-MSH). The melanocortin agonists contain the putative message sequence "His-Phe-Arg-Trp," and it has been well-documented that inversion of chirality of the Phe to DPhe results in a dramatic increase in melanocortin receptor potency. Herein, we report a tetrapeptide library, based upon the template Ac-His-DPhe-Arg-Trp-NH(2), consisting of 26 members that have been modified at the DPhe(7) position (alpha-MSH numbering) and pharmacologically characterized for agonist and antagonist activity at the mouse melanocortin receptors MC1R, MC3R, MC4R, and MC5R. The most notable results of this study include the identification of the tetrapeptide Ac-His-(pI)DPhe-Arg-Trp-NH(2) that is a full nanomolar agonist at the mMC1 and mMC5 receptors, a mMC3R partial agonist with potent antagonist activity (pA(2) = 7.25, K(i) = 56 nM) and, but unexpectedly, is a potent agonist at the mMC4R (EC(50) = 25 nM). This ligand possesses novel melanocortin receptor pharmacology, as compared to previously reported peptides, and is potentially useful for in vivo studies to differentiate MC3R vs MC4R physiological roles in animal models, such as primates, where "knockout" animals are not viable options. The DNal(2') substitution for DPhe resulted in a mMC3R partial agonist with antagonist activity (pA(2) = 6.5, K(i) = 295 nM) and a mMC4R (pA(2) = 7.8, K(i) = 17 nM) antagonist possessing 60- and 425-fold decreased potency, respectively, as compared with SHU9119 at these receptors. Examination of this DNal(2')-containing tetrapeptide at the F254S and F259S mutant mMC4Rs resulted in agonist activity of this mMC4R tetrapeptide antagonist, similar to that observed for the SHU9119 peptide, supporting our previously proposed hypothesis that the Phe 254 and 259 transmembrane six receptor residues are important for differentiating melanocortin sequence-based MC4R antagonists vs the agouti-related protein (AGRP) sequence-based antagonists.