Oxaliplatin transport mediated by organic cation/carnitine transporters OCTN1 and OCTN2 in overexpressing human embryonic kidney 293 cells and rat dorsal root ganglion neurons.
The organic cation/carnitine transporters OCTN1 and OCTN2 are related to other organic cation transporters (OCT1, OCT2, and OCT3) known for transporting oxaliplatin, an anticancer drug with dose-limiting neurotoxicity. In this study, we sought to determine whether OCTN1 and OCTN2 also transported oxaliplatin and to characterize their functional expression and contributions to its neuronal accumulation and neurotoxicity in dorsal root ganglion (DRG) neurons relative to those of OCTs. [(14)C]Oxaliplatin uptake, platinum accumulation, and cytotoxicity were determined in OCTN-overexpressing human embryonic kidney (HEK) 293 cells and primary cultures of rat DRG neurons. Levels of mRNA and functional activities of rat (r)Octns and rOcts in rat DRG tissue and primary cultures were characterized using reverse transcription-polymerase chain reaction and uptake of model OCT/OCTN substrates, including [(3)H]1-methyl-4-phenylpyridinium (MPP(+)) (OCT1-3), [(14)C]tetraethylammonium bromide (TEA(+)) (OCT1-3 and OCTN1/2), [(3)H]ergothioneine (OCTN1), and [(3)H]l-carnitine (OCTN2). HEK293 cells overexpressing rOctn1, rOctn2, human OCTN1, and human OCTN2 showed increased uptake and cytotoxicity of oxaliplatin compared with mock-transfected HEK293 controls; in addition, both uptake and cytotoxicity were inhibited by ergothioneine and L-carnitine. The uptake of ergothioneine mediated by OCTN1 and of L-carnitine mediated by OCTN2 was decreased during oxaliplatin exposure. rOctn1 and rOctn2 mRNA was readily detected in rat DRG tissue, and they were functionally active in cultured rat DRG neurons, more so than rOct1, rOct2, or rOct3. DRG neuronal accumulation of [(14)C]oxaliplatin and platinum during oxaliplatin exposure depended on time, concentration, temperature, and sodium and was inhibited by ergothioneine and to a lesser extent by L-carnitine but not by MPP(+). Loss of DRG neuronal viability during oxaliplatin exposure was inhibited by ergothioneine but not by L-carnitine or MPP(+). OCTN1 and OCTN2 both transport oxaliplatin and are functionally expressed by DRG neurons. OCTN1-mediated transport of oxaliplatin appears to contribute to its neuronal accumulation and treatment-limiting neurotoxicity more so than OCTN2 or OCTs.


Angiogenesis in the Septum and Inner Membrane of Refractory Chronic Subdural Hematomas: Consideration of Findings after Middle Meningeal Artery Embolization with Low-concentration n-butyl-2-cyanoacrylate.
The middle meningeal artery (MMA) is suggested to play an important role in the recurrence of chronic subdural hematomas (CSDHs). However, the exact mechanisms involved in the recurrence of CSDHs still remain unknown. For recurring CSDHs, MMA embolization is performed using low-concentration n-butyl-2-cyanoacrylate (NBCA) at our hospital. We report new findings and a discussion related to the mechanism of CSDH recurrence based on the imaging findings after MMA embolization, and cases that required craniotomy due to recurrence. The study included eight patients with recurrent CSDH, defined as ipsilateral hematoma re-enlargement within 3 months, and treated with MMA embolization. MMA embolization was performed successfully in all eight patients with no complications. Of the eight patients treated, one patient required craniotomy for the evacuation of CSDHs due to hematoma re-enlargement. NBCA casts were observed in the inner membrane of CSDHs in five of the eight patients by postoperative computed tomography scans. In the case that required craniotomy, the formation of neovasculatures was observed in the inner membrane of the CSDH. Our results suggested that angiogenesis also occurs in the septum and inner membrane through the MMA and the outer membrane of the CSDH. Additionally, these neovascular vessels may be involved in the recurrence of CSDH after MMA embolization. In a future larger study, it is necessary to elucidate in detail the vascular architecture of the CSDH membrane associated with the hematoma re-enlargement, and the effectiveness of MMA embolization that embolized to these peripheral neovascular vessels.


[Current status of high-resolution scrotal echography].
To review the main images that can be observed by ultrasound, trying to highlight what decision to take after their finding: exploratory surgery, orchyectomy, or follow-up. We performed a bibliographic review about the topic and incorporated our personal experience with scrotal ultrasound. Given that there is an extensive variety of anomalies that can be observed we divide them in liquid and solid lesions, and these in intra or extratesticular, plus a group of lesions that do not qualify in the previous groups. The development of high frequencies transducers (8-10 Mhz) and a greater experience today allows to diagnose not only testicular tumors but also a number of benign clinical entities, with enough specificity in some of them as to avoid surgery. Ultrasound is a simple test, painless, and can be repeated without trouble, so that it is the first test that should be ordered when managing any intra scrotal problem.


Plasma circRNA HIPK2 as a putative biomarker for the diagnosis and prediction of therapeutic effects in major depressive disorder.
Circular RNAs (circRNAs) are a prevalent type of non-coding RNAs exhibiting extensive expression in mammalian cells. Owing to their involvement in diverse pathophysiological mechanisms of major depressive disorder (MDD) and their inherent stability in peripheral blood, circRNAs have emerged as potential biomarkers of considerable significance. This study aimed to identify and validate circular RNA HIPK2 (circHIPK2) in MDD patients and to investigate its potential as a biomarker for the diagnosis and prognosis of MDD. Patients with MDD (n = 81) and healthy controls (HCs) (n = 48) were recruited for our study (October 2022 to June 2023). The expression of circHIPK2 in plasma was assessed using absolute quantitative polymerase chain reaction (qPCR). The expression of circHIPK2 in plasma of patients with MDD exhibited a significant increase compared to HCs. The circHIPK2 levels showed an area under the curve (AUC) of 0.796, corresponding to a specificity of 97.9% and a sensitivity of 60.4% in diagnosing MDD. Additionally, the rate of change in circHIPK2 over a 14-day period exhibited an AUC curve of 0.819, indicating its predictive value for antidepressive effects. CircHIPK2 could serve as a potential biomarker for diagnosing MDD and predicting therapeutic effects of MDD.


Free Will, Black Swans and Addiction.
The current dominant perspective on addiction as a brain disease has been challenged recently by Marc Lewis, who argued that the brain-changes related to addiction are similar to everyday changes of the brain. From this alternative perspective, addictions are bad habits that can be broken, provided that people are motivated to change. In that case, autonomous choice or "free will" can overcome bad influences from genes and or environments and brain-changes related to addiction. Even though we concur with Lewis that there are issues with the brain disease perspective, we also argue that pointing to black swans can be important, that is: there can be severe cases where addiction indeed tips over into the category of brain disease, but obviously that does not prove that every case of addiction falls into the disease category, that all swans are black. We argue that, for example, people suffering from Korsakoff's syndrome, can be described as having a brain disease, often caused by alcohol addiction. Moreover, the brain changes occurring with addiction are related to choice-behaviour (and the related notions of willed action), habit formation and insight, hence essential mental abilities to break the addiction. We argue for a more graded perspective, where both black swans (severe brain disease which makes recovery virtually impossible) and white swans (unaffected brain) are rare, and most cases of addiction come as geese in different shades of gray.


A study on the protective effect of Silybum marianum extract on hepatic ischemia-reperfusion injury.
The objective of the study was to study the protective effect of Silybum marianum extract on hepatic ischemia-reperfusion injury. Rats were randomly divided into five groups; namely Silybum marianum extract high-, medium-, and low-dose protection groups, model group and control group. Hepatic ischemia-reperfusion injury model was prepared. Serum or plasma AST, ALT, MDA, TNF-α, IL-1β, IL-6 levels were measured. The results revealed that after liver injury, AST, ALT, MDA, TNF-α, IL-1β, and IL-6 levels significantly increased in succession, showing significant differences. We concluded that inflammatory cytokines participate in liver injury and that Silybum marianum extract can reduce the production of inflammatory cytokines, and thus can have a protective effect on hepatic ischemia and reperfusion.


'The path through the unknown': the experience of being a relative of a dementia-suffering spouse or parent.
To examine the experiences of relatives of a spouse or parent who suffers from dementia and examines whether there are similarities or differences between these experiences. Dementia is an increasing illness in the world. Dementia affects not only the person with dementia but also the relatives. There is a lack of knowledge about the experience of being a relative to a dementia-suffering person. Twenty-one stories from relatives were included in this study, and these stories were analysed by employing Kirsti Malterud's method 'systematic text condensation'. The relatives were divided into four groups: sons, husbands, daughters and wives. Eight themes were identified in their stories, two in each of the four groups. From these eight themes, it was identified that they all experienced change, grief and negative personal sentiments. However, differences were also found, one of them being that the sons found it easier to adapt to new roles during the course of the illness, while the daughters found it more difficult. The husbands experienced being attacked by the people around them, while the wives were found to submit their dementia-suffering husbands to physical abuse. The wives also suffered from self-criticism. The similarities between the four groups are more significant than the differences. Sons, husbands, daughters and wives of a person suffering from dementia should be considered on an equal basis in terms of their experiences of grief, change and personal negative sentiments. Relatives play a significant role in the well-being of their parent or spouse suffering from dementia. Therefore, professionals need to focus on both relatives and patient when they meet a person with dementia in clinical practice.


Acute exercise stress reveals cerebrovascular benefits associated with moderate gains in cardiorespiratory fitness.
Elevated cardiorespiratory fitness improves resting cerebral perfusion, although to what extent this is further amplified during acute exposure to exercise stress and the corresponding implications for cerebral oxygenation remain unknown. To examine this, we recruited 12 moderately active and 12 sedentary healthy males. Middle cerebral artery blood velocity (MCAv) and prefrontal cortical oxyhemoglobin (cO(2)Hb) concentration were monitored continuously at rest and throughout an incremental cycling test to exhaustion. Despite a subtle elevation in the maximal oxygen uptake (active: 52±9 ml/kg per minute versus sedentary: 33±5 ml/kg per minute, P<0.05), resting MCAv was not different between groups. However, more marked increases in both MCAv (+28±13% versus +18±6%, P<0.05) and cO(2)Hb (+5±4% versus -2±3%, P<0.05) were observed in the active group during the transition from low- to moderate-intensity exercise. Collectively, these findings indicate that the long-term benefits associated with moderate increase in physical activity are not observed in the resting state and only become apparent when the cerebrovasculature is challenged by acute exertional stress. This has important clinical implications when assessing the true extent of cerebrovascular adaptation.


Clinical sensitivity and specificity of a real-time PCR assay for Campylobacter fetus subsp venerealis in preputial samples from bulls.
To determine clinical sensitivity and specificity of a quantitative real-time PCR (qRT-PCR) assay for Campylobacter fetus subsp venerealis (Cfv) in preputial samples of bulls. 313 beef bulls. Preputial samples were collected from 300 virgin bulls and 13 Cfv-infected bulls. Specificity of the qRT-PCR assay, determined on the basis of results for samples collected from virgin bulls, was compared with specificity of bacteriologic culture performed with transport enrichment medium (TEM). Sensitivity of the qRT-PCR assay, determined on the basis of results for multiple samples collected at weekly intervals from infected bulls, was compared with sensitivity of the direct fluorescent antibody test (DFAT), bacteriologic culture, and bacteriologic culture with TEM. Specificity was 85% for the qRT-PCR assay and 100% for bacteriologic culture; results were significantly different. Mean sensitivity was 85.4% for the qRT-PCR assay, 82.3% for direct culture in blood agar, 72.1% for the DFAT, 32.7% for direct culture in Skirrow agar, 30% for bacteriologic culture with TEM and blood agar, and 38.1% for bacteriologic culture with TEM and Skirrow agar. Differences in sensitivity among tests varied with ambient outdoor temperature. Repeated sampling significantly increased sensitivity of the qRT-PCR assay. Use of the qRT-PCR assay as a screening test on direct preputial samples had comparable sensitivity to bacteriologic culture, and repeated sampling improved sensitivity. Although improved performance of the qRT-PCR assay, compared with direct bacteriologic culture, was dependent on temperature, transport times that allow direct culture are unlikely under field conditions. The qRT-PCR assay would provide a fast and sensitive screening method for Cfv in bulls.


MYC/NBS1-Mediated DNA Damage Response is Involved in the Inhibitory Effect of Hydroxysafflor Yellow A on Glioma Cells.
The role of Hydroxysafflor Yellow A (HSYA) in glioma is less studied, this research determined the effect of HSYA on glioma cells. The expressions of MYC and NBS1 in glioma tissues were detected by bioinformatics analysis and verified by RT-qPCR. The target relationship between MYC and NBS1 was predicted by bioinformatics. After treating the cells with HSYA, silenced MYC, or overexpressed NBS1, the viability, apoptosis, proliferation, invasion, migration, and DNA damage of the glioma cells were detected by MTT, flow cytometry, colony formation, transwell, wound healing, and γH2AX immunofluorescence assays, respectively. IC50 of HSYA in glioma cells was analyzed by Probit regression analysis. The expressions of MYC, NBS1, factors related to migration, invasion, apoptosis, and DNA damage of the glioma cells were determined by Western blot or RT-qPCR. MYC and NBS1 were high-expressed in glioma, and NBS1 was targeted by MYC. HSYA and siRNA targeting MYC inhibited the cell viability, proliferation, invasion, migration, and induced the cell apoptosis of glioma cells. HSYA upregulated the expressions of MYC, γH2AX, E-Cadherin, Bax, and Cleaved-PARP1, stimulated the activation of NBS1, MRE11, RAD50, and ATM, and downregulated the expressions of N-Cadherin and Bcl2 in glioma cells. SiMYC decreased the IC50 of HSYA in the glioma cells, enhanced the sensitivity of glioma cells to HSYA, and inhibited the activation of NBS1 and ATM. NBS1 overexpression reversed the effect of siRNA targeting MYC on glioma cells. MYC silencing inhibited the DNA damage response via regulation of NBS1, leading to DNA repair deficiency, and subsequently enhanced the sensitivity of glioma cells to HSYA.


Sequential measurements of the tibial plateau angle in large-breed, growing dogs.
To determine the earliest age that canine tibial plateau angles (TPAs) can be reliably measured and determine whether TPAs change during long bone growth. 10 Labrador Retrievers and 20 Labrador Retriever-hound crossbreeds. Stifle joints were radiographed every 2 months from 8 weeks of age to radiographic closure of the tibial physes. Four examiners radiographically evaluated TPA, physeal closure status (ie, complete or incomplete) of the proximal and distal tibial physis, and whether anatomic TPA measurement landmarks were sufficiently visible (LSV) or insufficiently visible (LIV) for accurate measuring. Linear regression analysis was performed to detect change in TPAs over time. Mean ages with 95% confidence intervals (CIs) were determined for dogs with radiographs classified as LIV and LSV. TPAs did not change from 90 days of age to physeal closure. Mean age for dogs with radiographs classified as LIV was 70.2 days (95% CI, 68.12 to 72.28 days), with no dog with LIV radiographs over 81 days of age. Mean age for dogs with radiographs classified as LSV was 85.5 days (CI, 76.73 to 94.27 days). TPAs in Labrador Retrievers and Labrador Retriever-hound crossbreeds can be measured accurately after 90 days of age, and earlier attempts to measure result in falsely low TPA measurements. Measuring TPAs in growing dogs may allow earlier detection of premature physeal closures. As more is learned about the role of theTPA in cranial cruciate ligament injury, early treatment may be possible for growing dogs with cruciate ligament injuries and excessive tibial slope.


How to give away your cake and eat it too: Relinquishing control prompts reciprocal generosity.
Many resource allocations confer two rewards, but these rewards typically work in opposition to one another: Reputational rewards come to those who give and material rewards to those who receive. Eight studies reveal that abdicating a resource allocation decision-that is, giving away one's right to choose to someone else-may allow these two rewards to work in tandem. We found that people frequently abdicated to others, and abdication often prompted others to reciprocate by giving away the better of two items. This occurred in part because people perceived abdication to be generous; in fact, individuals who abdicated seemed nearly as generous as individuals who gave away the better item to begin with. Paradoxically, abdicating confers both the reputational benefits of giving and (often) the material benefits of getting. This finding has implications for everyday resource sharing behavior and as well as for theories of fairness and reciprocity. (PsycINFO Database Record (c) 2018 APA, all rights reserved).


Frequency specificity of chirp-evoked auditory brainstem responses.
This study examines the usefulness of the upward chirp stimulus developed by Dau et al. [J. Acoust. Soc. Am. 107, 1530-1540 (2000)] for retrieving frequency-specific information. The chirp was designed to produce simultaneous displacement maxima along the cochlear partition by compensating for frequency-dependent traveling-time differences. In the first experiment, auditory brainstem responses (ABR) elicited by the click and the broadband chirp were obtained in the presence of high-pass masking noise, with cutoff frequencies of 0.5, 1, 2, 4, and 8 kHz. Results revealed a larger wave-V amplitude for chirp than for click stimulation in all masking conditions. Wave-V amplitude for the chirp increased continuously with increasing high-pass cutoff frequency while it remains nearly constant for the click for cutoff frequencies greater than 1 kHz. The same two stimuli were tested in the presence of a notched-noise masker with one-octave wide spectral notches corresponding to the cutoff frequencies used in the first experiment. The recordings were compared with derived responses, calculated offline, from the high-pass masking conditions. No significant difference in response amplitude between click and chirp stimulation was found for the notched-noise responses as well as for the derived responses. In the second experiment, responses were obtained using narrow-band stimuli. A low-frequency chirp and a 250-Hz tone pulse with comparable duration and magnitude spectrum were used as stimuli. The narrow-band chirp elicited a larger response amplitude than the tone pulse at low and medium stimulation levels. Overall, the results of the present study further demonstrate the importance of considering peripheral processing for the formation of ABR. The chirp might be of particular interest for assessing low-frequency information.


Positive selection for bone morphogenetic protein receptor type-IB promotes differentiation and specification of human adipose-derived stromal cells toward an osteogenic lineage.
Adipose tissue represents an abundant and easily accessible source of multipotent cells that may serve as an excellent building block for tissue engineering. However, adipose-derived stromal cells (ASCs) are a heterogeneous group and subpopulations may be identified with enhanced osteogenic potential. Human ASC subpopulations were prospectively isolated based on expression of bone morphogenetic protein receptor type-IB (BMPR-IB). Unsorted, BMPR-IB(+), and BMPR-IB(-) cells were analyzed for their osteogenic capacity through histological staining and gene expression. To evaluate their in vivo osteogenic potential, critical-sized calvarial defects were created in immunocompromised mice and treated with unsorted, BMPR-IB(+), or BMPR-IB(-) cells. Healing was assessed using microcomputed tomography and pentachrome staining of specimens at 8 weeks. Increased osteogenic differentiation was noted in the BMPR-IB(+) subpopulation, as demonstrated by alkaline phosphatase staining at day 7 and extracellular matrix mineralization with Alizarin red staining at day 14. This was also associated with increased expression for osteocalcin, a late marker of osteogenesis. Radiographic analysis demonstrated significantly enhanced healing of critical-sized calvarial defects treated with BMPR-IB(+) ASCs compared with unsorted or BMPR-IB(-) cells. This was confirmed through pentachrome staining, which revealed more robust bone regeneration in the BMPR-IB(+) group. BMPR-IB(+) human ASCs have an enhanced ability to form bone both in vitro and in vivo. These data suggest that positive selection for BMPR-IB(+) and manipulation of the BMP pathway in these cells may yield a highly osteogenic subpopulation of cells for bone tissue engineering.


National trends in incidence, prevalence and disability-adjusted life years of invasive aspergillosis in Iran: a systematic review and meta-analysis.
Objectives: We aimed to study the epidemiology, prevalence, incidence, clinical manifestations, underlying diseases, treatments, outcomes, and societal impact through disability-adjusted life years (DALYs) of IA in Iran. Methods: A random-effect meta-analytic model was fitted to estimate the prevalence and incidence of IA in Iran. We also calculated DALYs. Results: Out of 79 published studies during the past 25 years from Iran, 23 met the inclusion criteria. A total of 2947 patients were included, of whom 396 (13.4%) patients were diagnosed with IA according to EORTC/MSG and ICU criteria. The main underlying condition for IA was hematologic disorders (39.4%). A. flavus 86 (43%) was the most common isolate. The pooled prevalence and incidence rates were 20.5 (95% CI 12.5 to 29.9) and 4.8 (95% CI 2.3-8.2) per 100,000 population, respectively. Total DALYs was estimated 164.13 per 100,000 population. YLLs constitute the majority of IA burden compared to YLDs (162.80 YLLs/100,000 population vs 1.33 YLDs per 100,000 population). The highest YLL rates were found in people aged 45-49 (62.9 YLLs/100,000 population) and 30-34 years (45.2 YLLs/100,000 population), respectively. Conclusion: This study indicates an increasing burden of IA in Iran, despite the extensive use of prophylaxis, challenging the public health, especially immunocompromised patients.


Effect of human atrial natriuretic peptide on blood pressure after sodium depletion in essential hypertension.
Human atrial natriuretic peptide was infused over four hours in three patients with essential hypertension. When the patients had a sodium intake of 200 mmol (mEq) daily an infusion of 0.5 micrograms atrial natriuretic peptide/min caused no significant change in blood pressure, whereas an infusion of 1.0 micrograms/min caused a gradual decrease in blood pressure and an increase in heart rate. After two to three hours of infusion with the higher dose two patients showed a sudden decrease in heart rate, with symptomatic hypotension. When the same patients had an intake of 50 mmol sodium daily their blood pressure was more sensitive to infusion of atrial natriuretic peptide; one patient again developed symptomatic hypotension, this time during an infusion of 0.5 micrograms/min. During all infusions distinct natriuresis occurred irrespective of whether blood pressure was affected. Prolonged, relatively low dose infusions of atrial natriuretic peptide can cause unwanted symptomatic hypotension. The effect on blood pressure is enhanced after sodium depletion, and blood pressure should be monitored carefully during longer infusions of atrial natriuretic peptide in patients with essential hypertension.


Upper airway motor outputs during vomiting versus swallowing in the decerebrate cat.
Swallowing and vomiting are antagonistic motor acts; nevertheless, vomiting can be immediately followed by swallowing. The purpose of this study was to clarify the interrelationship between these two behaviors, particularly in regard to comparing the upper airway motor patterns at the end of the expulsion phase with those during subsequent swallowing. Experiments were conducted using both paralyzed and non-paralyzed decerebrate cats, in which recordings were obtained either from upper airway muscles, the diaphragm and abdominal muscles or from the nerves that innervate those muscles. The activity patterns of most nerves recorded in paralyzed animals were consistent with the behavior recorded in non-paralyzed animals from the muscles innervated by those nerves, with the exception of the cricothyroid and stylopharyngeus muscles. Vomiting can be divided into a series of retches followed by expulsion, which itself can be further subdivided into three phases. The final stage of expulsion, characterized by burst-like exaggerated activity of the laryngeal elevator thyrohyoid and the pharyngeal constrictors, proved to be different from pharyngeal swallowing, as judged from differences in the spatio-temporal patterns of the upper airway motor outputs. However, post-vomiting swallowing activity was still observed even after total deafferentation of the laryngeal and pharyngeal areas in paralyzed animals. It is therefore likely that the central processes for vomiting and swallowing closely relate in generating these two behaviors.


Assessment of Growth Using Mandibular Canine Calcification Stages and Its Correlation with Modified MP3 Stages.
Orthodontic diagnosis and treatment planning for growing children must involve growth prediction, especially in the treatment of skeletal problems. Studies have shown that a strong association exists between skeletal maturity and dental calcification stages. The present study was therefore taken up to provide a simple and practical method for assessing skeletal maturity using a dental periapical film and standard dental X-ray machine, to compare the developmental stages of the mandibular canine with that of developmental stages of modified MP3 and to find out if any correlation exists, to determine if the developmental stages of the mandibular canine alone can be used as a reliable indicator for assessment of skeletal maturity. A total of 160 periapical radiographs (80 males and 80 females), of the mandibular right canine and the MP3 region was taken and assessed according to the Dermirjian's stages of dental calcification and the modified MP3 stages. The correlation between the developmental stages of MP3 and the mandibular right canine in male and female groups, is of high statistical significance (p = 0.001). The correlation coefficient between MP3 stages and developmental stages of mandibular canine and chronological age in male and females was found to be not significant. The correlation between the mandibular canine calcification stages and MP3 stages was found to be significant. The developmental stages of the mandibular canine could be used very reliably as a sole indicator for assessment of skeletal maturity.


Tribal gaps in Malaria in relation to KAP in East Godavari District, Andhra Pradesh.
To assess the level of GAPS in knowledge, attitudes and practices among tribal people regarding malaria disease. To assess the interassociation among knowledge, attitude and practices of scores. A cross-sectional study was performed in tribal areas of East Godavari District for a period of one year during 2013. Sample size was calculated as 1136. Multistage Random sampling method was used to identify the 4 PHC's in 4 directions of tribal areas and sample was equally distributed. In that study subjects were selected by using a simple Random method. Data were collected by using a pre-tested, semi-structured questionnaire and results were displayed. Setting: Tribal area of east Godavari district, Andhra Pradesh State, India. Participants: Study subjects were selected by simple random method. Knowledge-practice gap for source of mosquito breeding 5.99%, for the prevention of mosquito breeding sources and for the implementation of minor engineering measures it is 10.51%. For bush cutting, the knowledge practice gap is -20.07%. For potted plants, the knowledge practice gap is -8.45%. For broken pots, it is 68.93%. IRS is performed outside the house not inside the house. The KAP-GAP for screening is nil. Covering oneself with a blanket to prevent man-mosquito contact is 6.96%. In our study, we found Know-do gap or KAP-GAP in all aspects of prevention of mosquito breeding places and personal protective measures. Regarding the interassociation among knowledge, attitudes and practices, they were associated with each other with significant difference.


Social determinants of psychological distress in a nationally-representative sample of South African adults.
There is substantial evidence from developed countries that lower socioeconomic status (SES) is associated with increased occurrence of mental illness, and growing interest in the role of social support and social capital in mental health. However, there are few data on social determinants of mental health from low- and middle-income nations. We examined the association between psychological distress and SES, social support and bonding social capital in a nationally-representative sample of South African adults. As part of a national survey of mental health, a probability sample of 4,351 individuals was interviewed between 2002 and 2004. Non-specific psychological distress was measured using the Kessler K-10 scale. SES was assessed from an aggregate of household income, individual educational and employment status, and household material and financial resources. Social support, bonding social capital and traumatic life events were measured using multi-item scales. The mean age in the sample was 37 years and 76% of participants were black African. Measures of SES and social capital were inversely associated (p<0.001). Both recent and traumatic life events were more common among individuals with low levels of SES and social support. After adjusting for participant demographic characteristics and life events, high levels of psychological distress were most common among individuals with lower levels of SES and social capital. There was no independent association between levels of social support and psychological distress. The occurrence of recent life events appeared to partially mediate the association between SES and psychological distress (p=0.035) but not the association involving social capital (p=0.40). These data demonstrate persistent associations between levels of SES, social capital and psychological distress in South Africa. The increased frequency of recent life events appears to only partially explain higher levels of psychological distress among individuals of lower SES. Additional research is required to understand the temporality of this association as well as mechanisms through which SES and social capital influence mental health in low- and middle-income settings where high levels of poverty and trauma may contribute to excess burden of mental illness.


Textile Aging Characterization on New Generations of Explanted Commercial Endoprostheses: A Preliminary Study.
The goal of the present study was to assess the aging phenomena on second-generation textile endoprostheses (EPs) through explant analysis and to establish a preliminary classification of observed defects and material damages. From January 2011 to June 2016 110 second- and recent-generation EPs were collected as a part of a European collaborative retrieval program. The analysis focused on the first 41 consecutive commercial EPs collected between 2011 and 2014 and made from polyethylene terephthalate. Explants were submitted to a standardized evaluation protocol, which included data recording, eye-naked evaluation, cleaning of organic remnants, and structural analysis under numerical optical microscopy. Observations were reported using a classification based on 15 features evaluating the fabric, the stitches between the fabric and the stents, and the stents. The total surface area of the holes within the fabric was measured. EPs were implanted for thoracic and abdominal procedures in 12 and 29 cases, respectively. The mean ± SD duration of implantation was 34 ± 26 months (range 2 days-8 years). Sixty-four percent of the samples demonstrated at least one defect caused by compression damage potentially related to the insertion of the EP within the delivery system, which promoted holes and tears. Ninety-five percent of all EPs demonstrated at least one type of abrasion on the stitches. The degradation of the stitches and the number of ruptures increased with duration of implantation. Stent degradation was rare and consisted of corrosion and rupture. Cumulated holed surface area increased with time and was measured up to 13.5 mm2. Various aging-related phenomena on commercial textile EPs were identified and classified. Main damaging mechanisms were related to compression and abrasion leading to tears and holes in the fabric and rupture of stitches.


Evaluating Association Between Coronary Calcifications Visualized During Invasive Coronary Angiography With Total Mortality.
High calcium score is independently associated with a greater cardiac event rate. Using a large database of patients who underwent coronary angiography for clinical reasons, we evaluated the association between reported degree of coronary calcification with mortality and baseline risk factors. Using angiographic data of 1917 patients from 1993 to 1997, we studied any association between the locations of coronary calcium that were seen during coronary angiography with coronary artery risk factors. Furthermore, we correlated the locations of calcium with all cause mortality. A total of 1917 patients who underwent cardiac catheterization from 1993 to 1997 were studied. Total mortality was 22.9%. There was no association between the classic coronary risk factors (history of hypertension, hyperlipidemia, smoking, diabetes mellitus and family history) or race (White, Black, Hispanic, and Asian) with the occurrence of angiographic visible calcium in any location. Furthermore, we did not find any association between the locations of coronary calcium with all cause mortality. (All cause mortality occurred in 21.8% of patients with left main calcification vs. 23.3%, P = 0.63, in 24.6% of patients with left anterior descending artery calcification vs. 22.7%, P = 0.48, in 25.6% of patients with circumflex calcification vs. 23.1%, P = 0.52, in 25.7% of right coronary calcification vs. 22.7%, P = 0.47, in 24.6 of any coronary calcification vs. 22.5%, P = 0.4). Race, coronary risk factors, and all cause mortality are not associated with angiographic documented coronary calcification in any location in patients undergoing diagnostic coronary angiography.


Bacterial Adaptation during Chronic Respiratory Infections.
Chronic lung infections are associated with increased morbidity and mortality for individuals with underlying respiratory conditions such as cystic fibrosis (CF) and chronic obstructive pulmonary disease (COPD). The process of chronic colonisation allows pathogens to adapt over time to cope with changing selection pressures, co-infecting species and antimicrobial therapies. These adaptations can occur due to environmental pressures in the lung such as inflammatory responses, hypoxia, nutrient deficiency, osmolarity, low pH and antibiotic therapies. Phenotypic adaptations in bacterial pathogens from acute to chronic infection include, but are not limited to, antibiotic resistance, exopolysaccharide production (mucoidy), loss in motility, formation of small colony variants, increased mutation rate, quorum sensing and altered production of virulence factors associated with chronic infection. The evolution of Pseudomonas aeruginosa during chronic lung infection has been widely studied. More recently, the adaptations that other chronically colonising respiratory pathogens, including Staphylococcus aureus, Burkholderia cepacia complex and Haemophilus influenzae undergo during chronic infection have also been investigated. This review aims to examine the adaptations utilised by different bacterial pathogens to aid in their evolution from acute to chronic pathogens of the immunocompromised lung including CF and COPD.


The effectiveness of glucocorticoids in treating croup: meta-analysis.
To determine the effectiveness of glucocorticoid treatment in children with croup. Meta-analysis of randomised controlled trials that examine the effectiveness of glucocorticoid treatment in children with croup. Score on scale measuring severity of croup, use of cointerventions (adrenaline (epinephrine), antibiotics, or supplemental glucocorticoids), length of stay in accident and emergency or in hospital, and rate of hospitalisation. Twenty four studies met the inclusion criteria. Glucocorticoid treatment was associated with an improvement in the croup severity score at 6 hours with an effect size of -1.0 (95% confidence interval -1.5 to -0.6) and at 12 hours -1.0 (-1.6 to -0.4); at 24 hours this improvement was no longer significant (-1.0, -2.0 to 0.1). There was a decrease in the number of adrenaline treatments needed in children treated with glucocorticoids: a decrease of 9% (95% confidence interval 2% to 16%) among those treated with budesonide and of 12% (4% to 20%) among those treated with dexamethasone. There was also a decrease in the length of time spent in accident and emergency (-11 hours, 95% confidence interval -18 to 4 hours), and for inpatients hospital stay was reduced by 16 hours (-31 to 1 hour). Publication bias seems to play a part in these results. Dexamethasone and budesonide are effective in relieving the symptoms of croup as early as 6 hours after treatment. Fewer cointerventions are used and the length of time spent in hospital is decreased in patients treated with glucocorticoids.


Grouping of complex substances using analytical chemistry data: A framework for quantitative evaluation and visualization.
A detailed characterization of the chemical composition of complex substances, such as products of petroleum refining and environmental mixtures, is greatly needed in exposure assessment and manufacturing. The inherent complexity and variability in the composition of complex substances obfuscate the choices for their detailed analytical characterization. Yet, in lieu of exact chemical composition of complex substances, evaluation of the degree of similarity is a sensible path toward decision-making in environmental health regulations. Grouping of similar complex substances is a challenge that can be addressed via advanced analytical methods and streamlined data analysis and visualization techniques. Here, we propose a framework with unsupervised and supervised analyses to optimally group complex substances based on their analytical features. We test two data sets of complex oil-derived substances. The first data set is from gas chromatography-mass spectrometry (GC-MS) analysis of 20 Standard Reference Materials representing crude oils and oil refining products. The second data set consists of 15 samples of various gas oils analyzed using three analytical techniques: GC-MS, GC×GC-flame ionization detection (FID), and ion mobility spectrometry-mass spectrometry (IM-MS). We use hierarchical clustering using Pearson correlation as a similarity metric for the unsupervised analysis and build classification models using the Random Forest algorithm for the supervised analysis. We present a quantitative comparative assessment of clustering results via Fowlkes-Mallows index, and classification results via model accuracies in predicting the group of an unknown complex substance. We demonstrate the effect of (i) different grouping methodologies, (ii) data set size, and (iii) dimensionality reduction on the grouping quality, and (iv) different analytical techniques on the characterization of the complex substances. While the complexity and variability in chemical composition are an inherent feature of complex substances, we demonstrate how the choices of the data analysis and visualization methods can impact the communication of their characteristics to delineate sufficient similarity.


Whole blood GRHL2 expression as a prognostic biomarker in metastatic hormone-sensitive and castration-resistant prostate cancer.
As potent systemic therapies transition earlier in the prostate cancer disease course, molecular biomarkers are needed to guide optimal treatment selection for metastatic hormone-sensitive prostate cancer (mHSPC). The value of whole blood RNA to detect candidate biomarkers in mHSPC remains largely undefined. In this cohort study, we used a previously optimised whole blood reverse transcription polymerase chain reaction assay to assess the prognostic utility [measured by seven-month undetectable prostate-specific antigen (PSA) and time to castration-resistance (TTCR)] of eight prostate cancer-associated gene transcripts in 43 mHSPC patients. Transcripts with statistically significant associations (P<0.05) were further investigated in a metastatic castration-resistant prostate cancer (mCRPC) cohort (n=119) receiving contemporary systemic therapy, exploring associations with PSA >50% response (PSA50), progression-free survival (PFS) and overall survival (OS). Clinical outcomes were prospectively collected in a protected digital database. Kaplan-Meier estimates and multivariable Cox proportional-hazards models assessed associations between gene transcripts and clinical outcomes (mHSPC covariates: disease volume, docetaxel use and haemoglobin level; mCRPC covariates: prior exposure to chemotherapy or ARPIs, haemoglobin, performance status and presence of visceral disease). Follow-up was performed monthly during ARPI treatment, three-weekly during taxane chemotherapy, and three-monthly during androgen deprivation therapy (ADT) monotherapy. Serial PSA measurements were performed before each follow-up visit and repeat imaging was at the discretion of the investigator. Detection of circulating Grainyhead-like 2 (GRHL2) transcript was associated with poor outcomes in mHSPC and mCRPC patients. Detectable GRHL2 expression in mHSPC was associated with a lower rate of seven-month undetectable PSA levels (25% vs. 65%, P=0.059), and independently associated with shorter TTCR (HR 7.3, 95% CI: 1.5-36, P=0.01). In the mCRPC cohort, GRHL2 expression predicted significantly lower PSA50 response rates (46% vs. 69%, P=0.01), and was independently associated with shorter PFS (HR 3.1, 95% CI: 1.8-5.2, P<0.001) and OS (HR 2.9, 95% CI: 1.6-5.1, P<0.001). Associations were most apparent in patients receiving ARPIs. Detectable circulating GRHL2 was a negative prognostic biomarker in our mHSPC and mCRPC cohorts. These data support further investigation of GRHL2 as a candidate prognostic biomarker in metastatic prostate cancer, in addition to expanding efforts to better understand a putative role in therapeutic resistance to AR targeted therapies.


Histopathologic and immunohistochemical features in human skin after exposure to nitrogen and sulfur mustard.
N-methyl-2,2'-dichlorodiethylamine (HN2)is a topical chemotherapeutic agent used as therapy for cutaneous T-cell lymphomas (CTCL). Di(2-chloroethyl)sulfide (SM), and less often HN2, have been used as chemical weapons, with the skin being a principle target. The mechanisms by which these chemicals produce their therapeutic and toxic effects in skin, however, are not clearly defined. We exposed human skin explants to two doses of HN2 and SM. At 18 hours after exposure, histopathologic features were compared. In addition, immunohistochemical markers to basement membrane proteins were used to evaluate the effects of both chemicals on the basement membrane zone. Gross vesication was not seen. Pyknotic nuclei with or without dyskeratotic changes within epidermal keratinocytes were present at both doses. These changes varied more between skin specimens than they did between doses. Ballooning degeneration was more marked after SM exposures. Diffuse dermal-epidermal separation was present only at high-dose exposures and did not appear to correlate with the degree of changes locally in the overlying epidermis. Antibodies to laminin-5 showed decreased immunoreactivity after exposure to HN2 and SM. Immunoreactivity for laminin- was decreased to a lesser extent, and immunoreactivity for collagen IV and VII was unchanged. HN2 and SM produce similar histopathologic and immunohistochemical features after cutaneous exposure. These features suggest that part of mechanism of action of HN2 and SM is a direct effect on the basement membrane zone. Understanding the effects of HN2 and SM separate from their effect on DNA may be important in designing therapies and in advancing our understanding of the pathophysiologic changes induced by these chemicals when delivered topically.


Inhalation injury to tracheal epithelium in an ovine model of cotton smoke exposure. Early phase (30 minutes).
The purpose of this study was to evaluate lung cell injury during the acute phase of smoke inhalation injury. A group of 10 sheep were anesthetized with halothane and pancuronium followed by endotracheal intubation. In the first experiment 5 sheep were given air (sham group) and 5 were insufflated with cooled cotton smoke with a modified bee smoker. In the second part of our study (Experiment 2) the animals were insufflated with the following number of smoke breaths: 1 x 12 (n = 3); 2 x 12 (n = 4); 3 x 12 (n = 4) 4 x 12 (n = 4); and sham control (n = 1). After 30 min the animals were killed with KCl and the trachea prepared for scanning, transmission electron, and light microscopy. Our initial observation with scanning electron microscopy revealed a large amount of mucus on the surface of the epithelia. Numerous ciliated cells had been sloughed from the epithelium and were observed on the surface of the remaining ciliated cells. The sloughed cells were intact, and the cilia remained on the apical cell surface. Light and transmission electron microscopy revealed that most goblet cells were in the process of extruding mucus. The cytoplasm of goblet and basal cells appeared normal. Ciliated cells had a slightly vesiculated cytoplasm, and many were in the process of being sloughed from the epithelial surface. In these cells desmosomal attachment had been separated. The light microscope evaluation of the tracheal epithelium showed there was no dose-dependent effect between the four treatment groups.(ABSTRACT TRUNCATED AT 250 WORDS)


Does the wedge wing in the neck screw prevent cut-out failure in Ota/AO 31-A2 trochanteric fractures in elderly patients?
The purpose of this retrospective study was to evaluate the effects of Dyna Locking Trochanteric (DLT) nail (U & I Corporation 529-1, Yonghyun-dong, Uijungbu Kyunggi-Do, Korea 480-050) in 31-A1 and 31-A2 intertrochanteric femur fractures. Eighty-seven patients (63 female and 24 male, mean age 77 years) were treated. The mean duration of followup was 16.6 +/- 3.1 months (range, 12-24 months). There were 36 cases of 31-A1 and 51 cases of 31-A2 fractures. The duration of surgery, fluoroscopy screening time, the blood loss, the hospital stay, the tip-apex distance were significantly less in the 31-A1 fracture group compared with the 31-A2 fracture group (p < 0.001). Cut-out was observed in 13 patients (14.9%) above 80 years old with 31-A2 fractures. Especially in elderly patients with unstable 31-A2 fractures, the wedge wing on the neck screw does not work properly. Therefore further biomechanical investigations and also prospective clinical studies with larger series are needed to improve the device.


Refractory Hepatic Hydrothorax in Chronic Hepatitis C Controlled by Direct-acting Antivirals.
Hepatic hydrothorax is a transudative pleural effusion that complicates advanced liver cirrhosis. Patients refractory to medical treatment plus salt restriction and diuretics are considered to have refractory hepatic hydrothorax and may require transjugular intrahepatic portosystemic shunt (TIPS) or liver transplant. Successful antiviral therapy reduces the incidence of some complications of cirrhosis secondary to HCV infection. We report a case of hepatic hydrothorax in a 55-year-old female patient with HCV cirrhosis, which exhibited a spontaneous decrease in pleural effusion after direct antiviral agent (DAA) therapy. In cases of HCV cirrhosis, DAAs are worth administering before treatment by TIPS or liver transplantation.


Stable lead isotopes in environmental health with emphasis on human investigations.
There has been widespread use of stable lead isotopes in the earth sciences for more than 40 years focussed on the origin and age of rocks and minerals with lesser application in environmental investigations where the emphasis has been directed typically to the source of lead in environmental media such as air, water and soils. In contrast, the number of environmental health investigations focussed on humans (and primates) is limited in spite of the demonstrated utility of the approach in pioneering studies in the early 1970's. This paper reviews the status of lead isotopes in human investigations especially over the past 2 decades, the period over which most activity has taken place. Following a brief introduction to the method, examples are provided illustrating the use of lead isotopes in a wide spectrum of activities including sources and pathways of lead in diverse environments from urban to mining communities, various applications associated with pregnancy, the contribution of bone lead to blood lead including in the elderly, the half-life of lead in blood, and lead in bones and other media. A brief outline of critical research on non-human primates is also given. The lead isotope method is a powerful technique for tracing lead and could be employed more widely in human investigations.


Nutritional and health benefits of pulses.
Pulses (beans, peas, and lentils) have been consumed for at least 10 000 years and are among the most extensively used foods in the world. A wide variety of pulses can be grown globally, making them important both economically as well as nutritionally. Pulses provide protein and fibre, as well as a significant source of vitamins and minerals, such as iron, zinc, folate, and magnesium, and consuming half a cup of beans or peas per day can enhance diet quality by increasing intakes of these nutrients. In addition, the phytochemicals, saponins, and tannins found in pulses possess antioxidant and anti-carcinogenic effects, indicating that pulses may have significant anti-cancer effects. Pulse consumption also improves serum lipid profiles and positively affects several other cardiovascular disease risk factors, such as blood pressure, platelet activity, and inflammation. Pulses are high in fibre and have a low glycemic index, making them particularly beneficial to people with diabetes by assisting in maintaining healthy blood glucose and insulin levels. Emerging research examining the effect of pulse components on HIV and consumption patterns with aging populations indicates that pulses may have further effects on health. In conclusion, including pulses in the diet is a healthy way to meet dietary recommendations and is associated with reduced risk of several chronic diseases. Long-term randomized controlled trials are needed to demonstrate the direct effects of pulses on these diseases.


Epilepsy Risk Following Bariatric Surgery for Weight Loss.
A previous study reported finding that epilepsy risk is elevated after bariatric surgery for weight loss; however, this association has not been adequately explored. Our objectives were to (1) estimate the risk of epilepsy after bariatric surgery for weight loss relative to a nonsurgical cohort of patients with an obesity diagnosis and (2) identify epilepsy risk factors among bariatric surgery recipients. We conducted a population-based retrospective cohort study using linked health administrative databases in Ontario, Canada. Participants were accrued between July 1, 2010, and December 31, 2016, and followed until December 31, 2019. All Ontario residents aged 18 years and older who had bariatric surgery during the accrual period were eligible for inclusion in our exposed cohort. Patients hospitalized with a diagnosis of obesity and who did not have bariatric surgery during the accrual period were eligible for inclusion in our unexposed cohort. We excluded patients with a history of seizures, epilepsy, various seizure or epilepsy risk factors, psychiatric disorders, or drug or alcohol abuse/dependence. In our primary analysis, we used inverse probability of treatment weighting to control for confounding. A marginal Cox proportional hazards model was then used to estimate the risk of epilepsy associated with bariatric surgery. A Cox model was also used to identify epilepsy risk factors among exposed participants. The final sample included 16,958 exposed participants and 622,514 unexposed participants. After inverse probability of treatment weighting, the estimated rates of epilepsy were 50.1 and 34.1 per 100,000 person-years among those who did and did not have bariatric surgery, respectively. The hazard ratio for developing epilepsy after bariatric surgery was 1.45 (95% CI = 1.35, 1.56). Among participants who received bariatric surgery, stroke during follow-up increased epilepsy risk (HR = 14.03, 95% CI = 4.26, 46.25). In this study, we found that patients with a history of bariatric surgery were at increased risk of developing epilepsy. These findings suggest that epilepsy is a long-term risk associated with bariatric surgery for weight loss.


Inhibitory effect of 764-3 on Ca2+ uptake in rat brain synaptosomes.
In an attempt to examine the effect of 764-3, a purified component extracted from Radix Salviae Miltiorrhizae (RSM), on calcium channel, Ca2+ uptake in rat brain synaptosome stimulated by high-K+ and Glutamate (Glu) was investigated by using radioisotope labeled 45CaCl2. It was found that 764-3 could decrease the uptake of Ca2+ induced by high-K+ (60 mM) and 0.1 mM glutamate in a concentration-dependent manner. The maximum inhibiting rates were 45.22 +/- 4.88% and 37.58 +/- 4.52%, respectively. The results indicated that neuroprotective effect of RSM on cerebral ischemic injury may be related to its blocking effects on calcium channel.


The physics of MRI safety.
The main risks associated with magnetic resonance imaging (MRI) have been extensively reported and studied; for example, everyday objects may turn into projectiles, energy deposition can cause burns, varying fields can induce nerve stimulation, and loud noises can lead to auditory loss. The present review article is geared toward providing intuition about the physical mechanisms that give rise to these risks. On the one hand, excellent literature already exists on the practical aspect of risk management, with clinical workflow and recommendations. On the other hand, excellent technical articles also exist that explain these risks from basic principles of electromagnetism. We felt that an underserved niche might be found between the two, ie, somewhere between basic science and practical advice, to help develop intuition about electromagnetism that might prove of practical value when working around MR scanners. Following a wide-ranging introduction, risks originating from the main magnetic field, the excitation RF electromagnetic field, and switching of the imaging gradients will be presented in turn. 5 Technical Efficacy: 1 J. Magn. Reson. Imaging 2018;47:28-43.


Early exposure leads to repeatable preferences for male song in female zebra finches.
This study addressed the question of how early learning processes in females influence later preferences for a male trait. I tested whether exposure to song alone (of a male other than the father) was sufficient for inducing a stable (repeatable) preference in female zebra finches (Taeniopygia guttata) by limiting early exposure to tape tutoring. A group of controls heard no songs before also being tested in adulthood. Repeated tests for preferences for tutor or unfamiliar song were made, interspersed with additional tests involving new songs. Preferences were tested in an operant task where pecking of response keys led to song playback. Most females significantly preferred one of the two songs in a given test. In the first test, the relative preference for the tutor song was significantly higher for the tutored than for the control females. Subsequently, tutored females' preferences for the tutor song remained higher on average, but the two groups did not differ significantly. However, tutored, but not untutored females' preferences were highly repeatable between tests, suggesting that early exposure to song might lead to a consolidation ol choice behaviour, a previously unknown effect of early exposure to song in female songbirds.


The Helicobacter pylori ureC gene codes for a phosphoglucosamine mutase.
The function of UreC, the product of a 1,335-bp-long open reading frame upstream from the urease structural genes (ureAB) of Helicobacter pylori, was investigated. We present data showing that the ureC gene product is a phosphoglucosamine mutase. D. Mengin-Lecreulx and J. van Heijenoort (J. Biol. Chem. 271:32-39, 1996) observed that UreC is similar (43% identity) to the GlmM protein of Escherichia coli. Those authors showed that GlmM is a phosphoglucosamine mutase catalyzing interconversion of glucosamine-6-phosphate into glucosamine-1-phosphate, which is subsequently transformed into UDP-N-acetylglucosamine. The latter product is one of the main cytoplasmic precursors of cell wall peptidoglycan and outer membrane lipopolysaccharides. The present paper reports that, like its E. coli homolog glmM, the H. pylori ureC gene is essential for cell growth. It was known that growth of a lethal conditional glmM mutant of E. coli at a nonpermissive temperature can be restored in the presence of the ureC gene. We showed that complete complementation of the glmM mutant can be obtained with a plasmid overproducing UreC. The peptidoglycan content and the specific phosphoglucosamine mutase activity of such a complemented strain were measured; these results demonstrated that the ureC gene product functions as a phosphoglucosamine mutase. Homologs of the UreC and GlmM proteins were identified in Haemophilus influenzae, Mycobacterium leprae, Clostridium perfringens, Synechocystis sp. strain PCC6803, and Methanococcus jannaschii. Significant conservation of the amino acid sequence of these proteins in such diverse organisms suggests a very ancient common ancestor for the genes and defines a consensus motif for the phosphoglucosamine mutase active site. We propose renaming the H. pylori ureC gene the glmM gene.


Noncardiogenic mechanisms of right heart dysfunction.
Alterations in right ventricular (RV) performance are critical to the cardiac dysfunction witnessed in adult respiratory distress syndrome (ARDS), septic shock (SS), and as a consequence of positive end-expiratory pressure (PEEP) administration during mechanical ventilation. The authors review evidence for right heart dysfunction in these circumstances. In ARDS, an increase in RV afterload with the onset of pulmonary artery hypertension is the predominant factor promoting RV dysfunction. In SS, most investigators agree that a primary decrease in myocardial contractility is the major factor limiting RV performance. The application of PEEP during mechanical ventilation can potentiate alterations in RV preload, afterload, and/or contractility, all of which promote RV dysfunction and compromise left ventricular filling. As RV dysfunction may seriously affect global myocardial performance in all of these settings, the clinician must identify that RV function is impaired, discern the contributing mechanism, and select an appropriate therapeutic regimen targeted at addressing this predominant mechanism. Assessment and management strategies are described.


[Efficacy and adverse reactions of antipsychotics for neuropsychiatric symptoms in dementia: a systematic review].
To assess the efficacy and adverse reactions of typical and atypical antipsychotics in the treatment of neuropsychiatric symptoms in dementia, and to examine the evidence for the cerebrovascular events warning for atypical antipsychotics. Systematic review. Using Medline, Cinahl, PsyclNFO, Embase and the Cochrane central register of controlled trials (1980-2005), double-blind randomized controlled trials with intention-to-treat analysis were selected, which evaluated efficacy and adverse reactions of antipsychotics in the treatment of neuropsychiatric symptoms in dementia. The studies underwent a standardised validity assessment. After screening 950 studies, 14 studies on the effect of haloperidol, risperidone, olanzapine, quetiapine, tiapride, loxapine and perphenazine were selected. In 7 out of 10 studies, haloperidol, risperidone and olanzapine appeared to be more effective than placebo in the treatment of aggression and psychosis. Direct comparison between typical and atypical antipsychotics revealed no statistically significant difference. The most common adverse reactions were extrapyramidal symptoms and somnolence. These adverse reactions were less frequent with low-dose risperidone than with haloperidol or olanzapine, but risperidone and olanzapine were found to be associated with a higher risk of cerebrovascular events in two studies. The efficacy of typical and atypical antipsychotics is comparable, but only low-dose risperidone seems to be associated with fewer (extrapyramidal) side effects. The adverse reactions are inadequately described in the published data and consequently the warning of an increased risk of mortality could not be confirmed.


Job insecurity as a predictor of physiological indicators of health in healthy working women: an extension of previous research.
Job insecurity has been linked to different negative outcomes, such as negative work attitudes and health problems, with most studies including self-reported outcomes. Extending earlier research, the present study includes both self-reported and physiological indicators of health and sets out to investigate whether higher levels of job insecurity are related to higher levels of allostatic load, higher levels of morning cortisol, more physician-diagnosed symptoms of ill-health and poorer self-rated health. The study also investigated whether self-rated health mediated the relation between job insecurity and physiological outcomes. This was cross-sectionally studied in a cohort of Swedish women who participated in a large-scale longitudinal study focusing on life span development and adaptation. The results showed that job insecurity was related to self-rated health and morning cortisol, and, contrary to expectations, that job insecurity was unrelated to allostatic load and physician ratings, both directly and indirectly. The results indicate that, in healthy working women, job insecurity may be less detrimental to long-term physiological health than originally hypothesized.


Technical aspects and postoperative sequelae of spinal and epidural anesthesia: a prospective study of 3,230 orthopedic patients.
Major complications after spinal or epidural anesthesia are extremely rare. The occurrence of less serious and transient sequelae and complaints may be underestimated if there is no established organization for the systematic and continuous surveillance of patients after anesthesia. This study was designed to evaluate the possible relationship between various block-related occurrences and the intra- and postoperative side effects and complaints. This prospective study included 3,230 orthopedic patients operated on under neuraxial block. The block was performed by single-dose (single-shot spinal anesthesia [SPIN], 80.6%), continuous spinal anesthesia (CSA, 10.3%), or combined spinal and epidural anesthesia (CSE, 9.1%) technique. The patient position during surgery and a detailed description of block performance and equipment, as well as all intraoperative problems, were immediately recorded. Every patient was given a standardized questionnaire to be completed and returned after 1 week. The overall incidence of paresthesia was 12.8%, being most frequent during spinal catheter insertion with multiple attempts. Postoperative sensory disturbances (numbness, dysesthesia) occurred unrelated to paresthesia elicited by the puncture or catheterization. The failure rates (SPIN, 1.0%; CSA, 1.5%; and CSE, 1.0%) were quite low. Asystole with successful resuscitation, occurred in 1 patient with CSE but, overall, hemodynamic perturbations were more common with the continuous techniques. On the first postoperative day, decreased sensation of the skin was significantly more frequent after the continuous techniques, still present in 6.5% of SPIN patients, 6.1% of CSA patients, and 17.2% of CSE patients 1 week after surgery. These disturbances were also related to the use of a tourniquet and lateral position of the patient during surgery. The frequency of postdural puncture headache (PDPH) was similar for the different techniques (SPIN, 0.9%; CSA, 1.5%; and CSE, 1.7%). About one third of the patients reported strong postoperative pain on the day after the operation, and 5.6% continued to report this at 1 week. Although sensory changes were quite frequent, they were mild and transient. Rather than having been caused by the anesthetic technique, per se, a "nonanesthetic" reason (position, tourniquet, immobilization) should also be considered as their origin. Sensory disturbances, as well as strong pain for at least a week, were reported by several patients, and to become aware of these problems and improve the quality of treatment, a universal regional anesthesia follow-up system is recommended.


[A comparison of alpha 1-acid glycoprotein (AAG) concentration and disopyramide binding in Chinese and Japanese].
The mean concentration of alpha 1-acid glycoprotein (AAG) in plasma and disopyramine binding capacity to AAG (bound disopyramine/AAG) were compared between fourteen Chinese and sixteen Japanese subjects. As a results, the author observed that the mean AAG-concentrations in Chinese and in Japanese were 0.55 +/- 0.11 mg/ml and 0.64 +/- 0.13 mg/ml, respectively. When disopyramine concentration was at 5.0 micrograms/ml, the bound disopyramine/AAG was 7.3 +/- 1.3 micrograms/mg in Chinese and 6.5 +/- 1.3 micrograms/mg in Japanese. Unbound fractions of disopyramine in Chinese and Japanese were 0.23 +/- 0.05% and 0.19 +/- 0.07%, respectively. These results indicated that there was no significant difference in the AAG-concentration and the bound disopyramine/AAG between Chinese and Japanese subjects. In male subjects, however, the AAG-concentration and the bound disopyramine/AAG in Chinese (0.57 +/- 0.14 mg/ml and 7.47 +/- 1.78 micrograms/mg) were significantly different from those in Japanese (0.72 +/- 0.11 mg/ml and 5.84 +/- 0.76 micrograms/mg, p < 0.05), though unbound fraction of disopyramide was not found different between Chinese and Japanese subjects. In female, the AAG-concentration, the bound disopyramine/AAG and unbound fraction of disopyramide did not significantly differ between Chinese and Japanese subjects. These findings suggest a possible structural difference of monosaccharides in plasma-AAG between Chinese and Japanese male subjects.


[Analysis of the efficacy and influencing factors of sodium channel blockers in the treatment of focal epilepsy in infants under 6 months of age].
Objective: To analyze the efficacy and safety of the sodium channel blockers (SCB) antiseizure medication in the treatment of focal epilepsy in infants under 6 months of age. Methods: This was a case series study. Infants with focal epilepsy with onset within 6 months of age and treated with SCB attending the Department of Neurology of Beijing Children's Hospital from June 2016 to April 2022 were collected. The clinical data, auxiliary examinations, SCB application, efficacy, adverse reactions, and prognosis were analyzed retrospectively. Patients were grouped according to type of seizure and epileptic syndrome, age of onset and etiology. Chi square test and Fisher exact test were used to analyze the differences between groups statistically. Results: A total of 118 infants were enrolled, 65 males and 53 females, with an age of epilepsy onset of 56 (4, 114) days. Developmental and epileptic encephalopathy was diagnosed in 60 infants, 39 had self-limited neonatal and (or) infantile epilepsy, and 19 had non-syndromic focal epilepsy. Application of SCB: 106 used oxcarbazepine, 2 used lacosamide, 9 switched from oxcarbazepine to lacosamide or a combination of 2 SCB, and 1 used oxcarbazepine, lacosamide, and lamotrigine successively; oxcarbazepine was the first choice in 46 cases. The age at which SCB was applied was 103 (53, 144) days. The children were followed up for 6 months to 6 years. SCB was effective in 89 cases (75.4%), including 70 cases (59.3%) who achieved seizure freedom. The seizure-free rate was higher in the focal epilepsy only group than in the group with other seizure types (64.4% (65/101) vs. 4/17, χ²=9.99, P<0.05). The responder and seizure-free rates were all higher in the group with the onset age of >3-6 months than the group >1-3 months (84.4% (38/45) vs. 62.5% (20/32), 73.3% (33/45) vs. 46.9% (15/32), χ²=4.85 and 5.58, both P<0.05). With the exception of variants in the PRRT2 gene, those with variants in sodium or potassium channels had higher responder and seizure-free rates than those with variants in other genes(86.2% (25/29) vs. 45.5% (10/22), 62.1% (18/29) vs. 22.7% (5/22), χ²=9.65 and 7.82,both P<0.05). The most common adverse event was transient hyponatremia, which happened in 66 cases (55.9%). There were 9 cases of rash, which subsided in 6 cases after discontinuing oxcarbazepine and switching to lacosamide, and 7 cases of electrocardiogram abnormalities, which improved after withdrawing oxcarbazepine and changing to lacosamide in 1 case. Conclusion: SCB are effective and tolerable in the treatment of focal epilepsy in infants under 6 months of age, with better efficacy in patients with genetic variants of the sodium or potassium channel, focal seizures only, and seizure onset >3-6 months of age.


Respiratory Interventions, Hospital Utilization, and Clinical Outcomes of Persons with COPD and COVID-19.
Coronavirus disease 2019 (COVID-19) impacted outcomes of persons with chronic respiratory diseases such as chronic obstructive pulmonary disease (COPD). This study investigated the differences in respiratory interventions, hospital utilization, smoking status, and 30-day readmission in those with COPD and COVID-19 based on hospital survival status. A retrospective cross-sectional study was conducted from February 2020 to October 2020 and included persons with COPD and COVID-19 infection. We examined respiratory interventions, hospital utilization and outcomes, and 30-day hospital readmission. Chi-square test analysis was used to assess categorical variables, and t-test or Mann-Whitney was used to analyze continuous data based on normality. Ninety persons were included in the study, 78 (87%) were survivors. The most common comorbidity was hypertension 71 (78.9%) (p = 0.003). Twenty-two (24%) persons were intubated, from whom 12 (15%) survived (p < 0.001). There were 25 (32.1%) and 12 (100%), (p < 0.001) persons who required an ICU admission from the survivor and non-survivor groups, respectively. Among the survivor group, fifteen (19%) persons required 30-day hospital readmission. Persons with COPD and COVID-19 had a lower mortality rate (13%) compared to other studies in the early pandemic phase. Non-survivors had increased ICU utilization, endotracheal intubation, and more frequent application of volume control mode. Discharging survivors to long-term acute care facilities may reduce 30-day hospital readmissions.


Simultaneous characterization of rotational and translational diffusion of optically anisotropic particles by optical microscopy.
We probe the roto-translational Brownian motion of optically anisotropic particles suspended in water with a simple and straightforward optical microscopy experiment that does not require positional or rotational particle tracking. We acquire a movie of the suspension placed between two polarizing elements and we extract the translational diffusion coefficient D T and the rotational diffusion coefficient D R from the analysis of the temporal correlation properties of the spatial Fourier modes of the intensity fluctuations in the movie. Our method is successfully tested with a dilute suspension of birefringent spherical colloidal particles obtained by polymerizing an emulsion of droplets of liquid crystal in a nematic phase, whose roto-translational dynamics is found to be well described by theory. The simplicity of our approach makes our method a viable alternative to particle tracking and depolarized dynamic light scattering.


Identification of Translocation Inhibitors Targeting the Type III Secretion System of Enteropathogenic Escherichia coli.
Infections with enteropathogenic Escherichia coli (EPEC) cause severe diarrhea in children. The noninvasive bacteria adhere to enterocytes of the small intestine and use a type III secretion system (T3SS) to inject effector proteins into host cells to modify and exploit cellular processes in favor of bacterial survival and replication. Several studies have shown that the T3SSs of bacterial pathogens are essential for virulence. Furthermore, the loss of T3SS-mediated effector translocation results in increased immune recognition and clearance of the bacteria. The T3SS is, therefore, considered a promising target for antivirulence strategies and novel therapeutics development. Here, we report the results of a high-throughput screening assay based on the translocation of the EPEC effector protein Tir (translocated intimin receptor). Using this assay, we screened more than 13,000 small molecular compounds of six different compound libraries and identified three substances which showed a significant dose-dependent effect on translocation without adverse effects on bacterial or eukaryotic cell viability. In addition, these substances reduced bacterial binding to host cells, effector-dependent cell detachment, and abolished attaching and effacing lesion formation without affecting the expression of components of the T3SS or associated effector proteins. Moreover, no effects of the inhibitors on bacterial motility or Shiga-toxin expression were observed. In summary, we have identified three new compounds that strongly inhibit T3SS-mediated translocation of effectors into mammalian cells, which could be valuable as lead substances for treating EPEC and enterohemorrhagic E. coli infections.


Edge Factors: Scientific Frontier Positions of Nations.
A key decision in scientific work is whether to build on novel or well-established ideas. Because exploiting new ideas is often harder than more conventional science, novel work can be especially dependent on interactions with colleagues, the training environment, and ready access to potential collaborators. Location may thus influence the tendency to pursue work that is close to the edge of the scientific frontier in the sense that it builds on recent ideas. We calculate for each nation its position relative to the edge of the scientific frontier by measuring its propensity to build on relatively new ideas in biomedical research. Text analysis of 20+ million publications shows that the United States and South Korea have the highest tendencies for novel science. China has become a leader in favoring newer ideas when working with basic science ideas and research tools, but is still slow to adopt new clinical ideas. Many locations remain far behind the leaders in terms of their tendency to work with novel ideas, indicating that the world is far from flat in this regard.


Crizotinib changes the metabolic pattern and inhibits ATP production in A549 non-small cell lung cancer cells.
Crizotinib, an inhibitor of the hepatocyte growth factor receptor oncogene, has been studied extensively regarding its antitumor and clinically beneficial effects in non-small cell lung cancer (NSCLC). However, crizotinib's effects on cancer cell energy metabolism, which is linked with tumor proliferation and migration, in NSCLC are unclear. Therefore, the present study focused on crizotinib's effect on NSCLC glucose metabolism. Crizotinib's effects on glucose metabolism, proliferation, migration and apoptosis in A549 cells were explored. Several other inhibitors, including 2-DG, rotenone and MG132, were used to define the mechanism of action in further detail. Data showed that crizotinib treatment reduced A549 cell viability, increased glucose consumption and lactate production, while decreased mitochondrial transmembrane potential (Δψm) and ATP production. Crizotinib treatment, combined with rotenone and MG132 treatment, further inhibited ATP production and Δψm and increased reactive oxygen species content. However, crizotinib did not suppress cell proliferation, migration, ATP production, Δψm or mitochondrial-related apoptosis signals further following 2-DG-mediated inhibition of glycolysis. These results indicated that crizotinib induced low mitochondrial function and compensatory high anaerobic metabolism, but failed to maintain sufficient ATP levels. The alternation of metabolic pattern and insufficient ATP supply may serve important roles in the metabolic antitumor mechanism of crizotinib in A549 cells.


Malnutrition Risk, Physical Function Decline and Disability in Middle-Aged and Older Adults Followed Up in the Singapore Longitudinal Ageing Study.
Malnutrition and population ageing are doubly global healthcare challenges. It is widely recognized that disability is a major contributor to malnutrition among older people, but the importance of reducing malnutrition risk (MR) for disability prevention is given little attention. We investigated the association between MR and risk of incident disability and functional decline among community-dwelling older persons. Prospective cohort study of community-dwelling older adults aged 55 and above (Singapore Longitudinal Ageing Study) with 3-5 years follow-up. MR was measured at baseline using an appropriate and validated index (ENIGMA); physical and functional measures at baseline and follow-up included instrumental and basic activity of daily living (ADL), knee extension (KES), gait velocity (GV), timed up-and-go (TUG). Compared to low MR, mild-moderate MR (OR = 1.43, 95% CI = 1.02-2.01) and severe MR (OR=1.74, 95% CI=1.10-2.74) were associated with higher risks of incident disability. Severe MR was associated with functional decline (OR = 1.69, 95% CI = 1.11-2.57). Estimates were adjusted for demographic, social, lifestyle behaviour and health variables. In particular, eating difficulty was associated with incident disability and functional decline, and low lymphocyte with functional decline. Severe MR and low albumin were associated with 37% worse GV and 126% worse TUG declines; consuming few vegetables or fruits with 34% worse KES decline; polypharmacy with 56% worse TUG decline. Malnutrition risk increased the risk of incident disability and functional decline in non-disabled persons, it worsened physical performance declines. Further studies should investigate the effectiveness of nutritional interventions in reducing the risk of disability among older people.


Total Ankle Arthroplasty Survivorship, Complication, and Revision Rates in Patients Younger Than 55 Years.
Total ankle replacement has become a viable alternative to ankle arthrodesis in the surgical management of advanced ankle arthritis. Total ankle replacement has generally been reserved for patients who are older and for those who will have a lower demand on the replacement. The purpose of the current study is to review patient outcomes, complications, and implant survival in patients younger than 55 years who underwent total ankle replacement at a single institution. A single-center chart and radiographic review was performed of consecutive patients who underwent total ankle replacement for treatment of end-stage ankle arthritis. All surgeries were performed by 1 of 5 fellowship-trained foot and ankle surgeons at a single institution. A total of 51 patients met inclusion criteria with a mean follow-up of 31.2 months (SD = 16.2). Implant survival was 94%, There were 7 major complications (13%) requiring an unplanned return to the operating room and 8 minor complications (15%) that resolved with conservative care. The results of this study show that total ankle replacement is a viable treatment option for patients younger than 55 years.Levels of Evidence: A retrospective case series.


Risk Factors for Hemorrhagic Transformation in Patients with Acute Middle Cerebral Artery Infarction.
Hemorrhagic transformation (HT) after acute ischemic stroke (AIS) can be seen at any time following ischemic stroke. Although HT usually occurs as a complication of antithrombotic, anticoagulant, or thrombolytic treatments, it can also occur spontaneously. We aimed to investigate the occurrence of early HT and its relevant risk factors in patients diagnosed with acute middle cerebral artery (MCA) infarction who were not treated with thrombolytic agents. We recruited 171 patients with acute MCA infarction between January 2011 and July 2012 who were not treated with thrombolytic agents and were suitable to our inclusion criteria. Controlled neuroimaging was performed immediately in patients with deterioration, otherwise on day 7 following stroke. All patients were investigated for AIS risk factors and biochemical analyses were performed. Patients with HT in controlled neuroimaging were grouped both clinically (i.e., symptomatic or asymptomatic) and radiologically, according to "European Cooperative Acute Stroke Radiological Study" (ECASS), and risk factors were examined. We enrolled 171 patients [94 men (55%) and 77 women (45%)] in the study. HT developed in 37 patients (21.63%). In terms of risk factor analysis, the most frequent etiological factor was atherosclerosis in AIS patients (50.3%). National Institutes of Health Stroke Scale scores were significantly higher both in sHT patients according to asHT patients and in HT patients on day 7 compared with their initial scores. Serum low-density lipoprotein (LDL-C), triglycerides (TG), and total cholesterol (TC) levels were significantly lower in patients with HT (p<.001). HT is a major complication in AIS that considerably increases the morbidity and mortality. To reduce the occurrence of HT, risk factors for each patient population should be determined. Acute thrombolytic therapy should be used cautiously in high-risk patients, and appropriate alternative therapies should be revised in them.


Mannheim Peritonitis Index (MPI) and elderly population: prognostic evaluation in acute secondary peritonitis.
Acute Secondary Peritonitis due to abdominal visceral perforation is characterized by high mortality and morbidity risk. Risk stratification allows prognosis prediction to adopt the best surgical treatment and clinical care support therapy. In Western countries elderly people represent a significant percentage of population Aim. Evaluation of Mannheim Peritonitis Index (MPI) and consideration upon old people. Retrospective study on 104 patients admitted and operated for "Acute Secondary Peritonitis due to visceral perforation". MPI was scored. In our study we want to demonstrate efficacy of MPI and the possibility to consider older age an independent prognostic factor. Mortality was 25.96%. Greatest sensitivity and specificity for the MPI score as a predictor of mortality was at the score of 20. MPI score of <16 had 0.15 times lower risk of mortality compared to patients with MPI score 17 - 21 and 0.61 lower than patients with MPI >22. Patients with MPI score 17-21 had 0.46 times lower risk of mortality compared to patients with MPI score >21. In the group of patients with MPI score of >20 the mortality rate was 48.5% for patients older than 80 years old and 12.1% for younger patients (p < 0.005); in the group with MPI score of < 20 mortality rate was respectively 8.4% and 1.4% (p < 0.005). Data confirm the accuracy of the test. MPI score and age over 80 years old resulted independent predictors of mortality at multivariate analysis.


Do Verbal Interactions with Infants During Electronic Media Exposure Mitigate Adverse Impacts on their Language Development as Toddlers?
The goal of this study was to determine whether verbal interactions between mothers and their 6-month-old infants during media exposure ('media verbal interactions') might have direct positive impacts, or mitigate any potential adverse impacts of media exposure, on language development at 14 months. For 253 low-income mother-infant dyads participating in a longitudinal study, media exposure and media verbal interactions were assessed using 24-hour recall diaries. Additionally, general level of cognitive stimulation in the home [StimQ] was assessed at 6 months and language development [Preschool Language Scale-4] was assessed at 14 months. Results suggest that media verbal interactions play a role in the language development of infants from low-income, immigrant families. Evidence showed that media verbal interactions moderated adverse impacts of media exposure found on 14-month language development, with adverse associations found only in the absence the these interactions. Findings also suggest that media verbal interactions may have some direct positive impacts on language development, in that media verbal interactions during the co-viewing of media with educational content (but not other content) were predictive of 14-month language independently of overall level of cognitive stimulation in the home.


Placental transforming growth factor-beta is a downstream mediator of the growth arrest and apoptotic response of tumor cells to DNA damage and p53 overexpression.
The p53 tumor suppressor gene and members of the transforming growth factor-beta (TGF-beta) superfamily play central roles in signaling cell cycle arrest and apoptosis (programmed cell death) in normal development and differentiation, as well as in carcinogenesis. Here we describe a distantly related member of the TGF-beta superfamily, designated placental TGF-beta (PTGF-beta), that is up-regulated in response to both p53-dependent and -independent apoptotic signaling events arising from DNA damage in human breast cancer cells. PTGF-beta is normally expressed in placenta and at lower levels in kidney, lung, pancreas, and muscle but could not be detected in any tumor cell line studied. The PTGF-beta promoter is activated by p53 and contains two p53 binding site motifs. Functional studies demonstrated that one of these p53 binding sites is essential for p53-mediated PTGF-beta promoter induction and specifically binds recombinant p53 in gel mobility shift assays. PTGF-beta overexpression from a recombinant adenoviral vector (AdPTGF-beta) led to an 80% reduction in MDA-MB-468 breast cancer cell viability and a 50-60% reduction in other human breast cancer cell lines studied, including MCF-7 cells, which are resistant to growth inhibition by recombinant wild-type p53. Like p53, PTGF-beta overexpression was seen to induce both G(1) cell cycle arrest and apoptosis in breast tumor cells. These results provide the first evidence for a direct functional link between p53 and the TGF-beta superfamily and implicate PTGF-beta as an important intercellular mediator of p53 function and the cytostatic effects of radiation and chemotherapeutic cancer agents.


The HPV16 E6 binding protein Tip-1 interacts with ARHGEF16, which activates Cdc42.
Guanidine exchange factor (GEF)-catalysed activation of Rho proteins such as Cdc42 has been shown to have a crucial role in cellular transformation, malignant progression and invasion. We have previously shown that the HPV16 E6 oncoprotein binds to the PDZ domain protein Tax-interacting-protein 1 (Tip-1) and we now report identification and functional analysis of a novel Tip-1 binding GEF. Yeast two-hybrid, in vitro pull-down, site-directed mutagenesis, semiquantitative PCR, co-immunoprecipitation and western blotting were used to identify/confirm novel Tip-1 binding partners and analyse cellular expression levels. In vitro kinetic analyses of recombinant proteins, siRNA gene silencing and in cell assays were used to measure Rho protein activation. Tax-interacting-protein 1 was shown to interact with ARHGEF16 by its carboxyl PDZ binding motif. Levels of ARHGEF16 were increased in transformed and immortalised cells expressing ectopic HPV16 E6 and Cdc42 was co-immunoprecipitated by ARHGEF16 in the presence of high-risk HPV E6. In vitro kinetic analysis confirmed that recombinant ARHGEF16 activates Cdc42 and this was increased by the addition of recombinant Tip-1 and E6. Cells expressing HPV16 E6 had higher levels of Cdc42 activation, which was decreased by siRNA silencing of either Tip-1 or ARHGEF16. These data suggest that HPV16 E6, Tip-1 and ARHGEF16 may cooperate to activate Cdc42 and support a potential link between the expression of HPV16 E6 and Cdc42 activation.


Specificity and evolutionary conservation of the Escherichia coli RNA pyrophosphohydrolase RppH.
Bacterial RNA degradation often begins with conversion of the 5'-terminal triphosphate to a monophosphate by the RNA pyrophosphohydrolase RppH, an event that triggers rapid ribonucleolytic attack. Besides its role as the master regulator of 5'-end-dependent mRNA decay, RppH is important for the ability of pathogenic bacteria to invade host cells, yet little is known about how it chooses its targets. Here, we show that Escherichia coli RppH (EcRppH) requires at least two unpaired nucleotides at the RNA 5' end and prefers three or more such nucleotides. It can tolerate any nucleotide at the first three positions but has a modest preference for A at the 5' terminus and either a G or A at the second position. Mutational analysis has identified EcRppH residues crucial for substrate recognition or catalysis. The promiscuity of EcRppH differentiates it from its Bacillus subtilis counterpart, which has a strict RNA sequence requirement. EcRppH orthologs likely to share its relaxed sequence specificity are widespread in all classes of Proteobacteria, except Deltaproteobacteria, and in flowering plants. By contrast, the phylogenetic range of recognizable B. subtilis RppH orthologs appears to be restricted to the order Bacillales. These findings help to explain the selective influence of RppH on bacterial mRNA decay and show that RppH-dependent degradation has diversified significantly during the course of evolution.


Intermittent complete androgen blockade in metastatic prostate cancer.
Intermittent hormonal treatment of prostate cancer was first developed based upon experimental study results. Using the Shionogi mouse breast cancer model, it was shown that the tumor grows rapidly in the presence of androgens, then undergoes apoptotic regression when androgens are removed. This apoptotic potential can be reinduced several times by cyclic replacement and withdrawal of androgens. These results led to the concept being evaluated in clinical trials. In most of the clinical studies a protocol is used in which the patient receives 36 weeks of androgen deprivation. For those patients whose prostate-specific antigen (PSA) drops to less than 4 ng/ml within 32 weeks of therapy, the androgen withdrawal is stopped at 36 weeks, and not reintroduced until PSA increases to 20 ng/ml. This cycle is then repeated until the patient's tumor becomes hormone-sensitive. Akakura et al. in 1993, reported on 7 patients who had a total of 12 episodes off-hormone therapy after achieving PSA complex remission, with further response after each re-exposure. Bruchovsky et al. in 1997, reported on 47 patients who entered a study of intermittent hormone therapy to evaluate the effect of cyclic withdrawal and replacement therapy in 14 D2, 10 D, 19 C, 2 B2, 2 A2 patients. Treatment was initiated with combined androgen blockade and continued for at least 6 months until a serum PSA nadir was observed. The first two treatment cycles lasted 73 and 75 weeks, with a mean time-off therapy of 30 and 33 weeks and an overall mean percentage time-off therapy of 41 and 45%. Serum testosterone returned to the normal range within 8 weeks (range: 1-26 weeks) of stopping treatment. The off-treatment period in both cycles was associated with an improvement in sense of well-being, and the recovery of libido and potency in the men who reported normal or near-normal sexual function before the start of therapy. In 7 patients with stage D2 disease, the cancer progressed to an androgen-independent state. The mean and median times to progression were 128 and 108 weeks. Seven patients died, one from a non-cancer-related illness, with mean and median overall survival times of 210 and 166 weeks. These studies demonstrated that the androgen-dependent state of prostate cancer can be maintained during a course of intermittent androgen suppression, supporting the possibility of multiple apoptotic regressions under well-regulated conditions. Oliver et al. in 1997, conducted a retrospective study of 20 patients and concluded that intermittent androgen deprivation reduced induction of hormone-resistant prostate cancer, with no acute or major risk associated with the use of intermittent androgen suppression. More clinical studies are required to clarify the indication for intermittent hormone therapy and evaluate improvement in quality of life and survival. In the future, approaches to the improvement of therapeutic apoptosis could include intermittent hormone therapy, associated with additive cytotoxic therapeutic strategies.


A Statistical Procedure for Genome-Wide Detection of QTL Hotspots Using Public Databases with Application to Rice.
Genome-wide detection of quantitative trait loci (QTL) hotspots underlying variation in many molecular and phenotypic traits has been a key step in various biological studies since the QTL hotspots are highly informative and can be linked to the genes for the quantitative traits. Several statistical methods have been proposed to detect QTL hotspots. These hotspot detection methods rely heavily on permutation tests performed on summarized QTL data or individual-level data (with genotypes and phenotypes) from the genetical genomics experiments. In this article, we propose a statistical procedure for QTL hotspot detection by using the summarized QTL (interval) data collected in public web-accessible databases. First, a simple statistical method based on the uniform distribution is derived to convert the QTL interval data into the expected QTL frequency (EQF) matrix. And then, to account for the correlation structure among traits, the QTL for correlated traits are grouped together into the same categories to form a reduced EQF matrix. Furthermore, a permutation algorithm on the EQF elements or on the QTL intervals is developed to compute a sliding scale of EQF thresholds, ranging from strict to liberal, for assessing the significance of QTL hotspots. With grouping, much stricter thresholds can be obtained to avoid the detection of spurious hotspots. Real example analysis and simulation study are carried out to illustrate our procedure, evaluate the performances and compare with other methods. It shows that our procedure can control the genome-wide error rates at the target levels, provide appropriate thresholds for correlated data and is comparable to the methods using individual-level data in hotspot detection. Depending on the thresholds used, more than 100 hotspots are detected in GRAMENE rice database. We also perform a genome-wide comparative analysis of the detected hotspots and the known genes collected in the Rice Q-TARO database. The comparative analysis reveals that the hotspots and genes are conformable in the sense that they co-localize closely and are functionally related to relevant traits. Our statistical procedure can provide a framework for exploring the networks among QTL hotspots, genes and quantitative traits in biological studies. The R codes that produce both numerical and graphical outputs of QTL hotspot detection in the genome are available on the worldwide web http://www.stat.sinica.edu.tw/chkao/.