Experimental investigation of a moving averaging algorithm for motion perpendicular to the leaf travel direction in dynamic MLC target tracking.
In dynamic multileaf collimator (MLC) motion tracking with complex intensity-modulated radiation therapy (IMRT) fields, target motion perpendicular to the MLC leaf travel direction can cause beam holds, which increase beam delivery time by up to a factor of 4. As a means to balance delivery efficiency and accuracy, a moving average algorithm was incorporated into a dynamic MLC motion tracking system (i.e., moving average tracking) to account for target motion perpendicular to the MLC leaf travel direction. The experimental investigation of the moving average algorithm compared with real-time tracking and no compensation beam delivery is described. The properties of the moving average algorithm were measured and compared with those of real-time tracking (dynamic MLC motion tracking accounting for both target motion parallel and perpendicular to the leaf travel direction) and no compensation beam delivery. The algorithm was investigated using a synthetic motion trace with a baseline drift and four patient-measured 3D tumor motion traces representing regular and irregular motions with varying baseline drifts. Each motion trace was reproduced by a moving platform. The delivery efficiency, geometric accuracy, and dosimetric accuracy were evaluated for conformal, step-and-shoot IMRT, and dynamic sliding window IMRT treatment plans using the synthetic and patient motion traces. The dosimetric accuracy was quantified via a tgamma-test with a 3%/3 mm criterion. The delivery efficiency ranged from 89 to 100% for moving average tracking, 26%-100% for real-time tracking, and 100% (by definition) for no compensation. The root-mean-square geometric error ranged from 3.2 to 4.0 mm for moving average tracking, 0.7-1.1 mm for real-time tracking, and 3.7-7.2 mm for no compensation. The percentage of dosimetric points failing the gamma-test ranged from 4 to 30% for moving average tracking, 0%-23% for real-time tracking, and 10%-47% for no compensation. The delivery efficiency of moving average tracking was up to four times higher than that of real-time tracking and approached the efficiency of no compensation for all cases. The geometric accuracy and dosimetric accuracy of the moving average algorithm was between real-time tracking and no compensation, approximately half the percentage of dosimetric points failing the gamma-test compared with no compensation.


Immune response genes in the post-Q-fever fatigue syndrome, Q fever endocarditis and uncomplicated acute primary Q fever.
The influence of immune response gene variations on the development of chronic complications of Q fever is presently unclear. To compare the frequencies of allelic polymorphisms in immune response genes in different Q fever patient groups. Genetic association study. We measured the frequencies of immune response gene variants in: (i) an expanded group of 31 post-Q-fever fatigue patients (QFS); (ii) 22 Q fever endocarditis patients (QFE); and (iii) 22 patients who made an uncomplicated recovery from their initial attack of primary acute Q fever, comparing them with various standard control panels from the general population. There were significant differences between the three Q fever groups. QFS patients differed from both QFE and uncomplicated patients and controls in the frequency of carriage of HLA-DRB1*11 and of the 2/2 genotype of the interferon-gamma intron1 microsatellite. Carriage of the HLA DRB1*11 allele was associated with reduced interferon-gamma and IL-2 responses from PBMC stimulated with ligand in short-term culture. QFE showed differences in the IL-10 promoter microsatellites R and G and had higher frequencies of the TNF-alpha receptor II 196R polymorphism. Q fever patients who had made an uncomplicated recovery differed from those with QFS or QFE, but were not significantly different in allelic frequencies to the control panels. These immunogenetic differences support the concept of different immune states in chronic Q fever, determined by genetic variations in host immune responses, rather than by solely properties of Coxiella burnetii.


Photovoltaic effect in individual asymmetrically contacted lead sulfide nanosheets.
Solution-processable, two-dimensional semiconductors are promising optoelectronic materials which could find application in low-cost solar cells. Lead sulfide nanocrystals raised attention since the effective band gap can be adapted over a wide range by electronic confinement and observed multi-exciton generation promises higher efficiencies. We report on the influence of the contact metal work function on the properties of transistors based on individual two-dimensional lead sulfide nanosheets. Using palladium we observed mobilities of up to 31 cm(2) V(-1) s(-1). Furthermore, we demonstrate that asymmetrically contacted nanosheets show photovoltaic effect and that the nanosheets' height has a decisive impact on the device performance. Nanosheets with a thickness of 5.4 nm contacted with platinum and titanium show a power conversion efficiency of up to 0.94% (EQE 75.70%). The results underline the high hopes put on such materials.


Regulation of oncogenic genes by MicroRNAs and pseudogenes in human lung cancer.
Lung cancer is one of the most common mortal cancer types both for men and women. Several different biomarkers have been analyzed to reveal lung cancer prognosis pathways for developing efficient therapeutics and diagnostic agents. microRNAs (miRNAs) and pseudogenes are critical biomarkers in lung cancer and alteration of their expression levels has been identified in each step of lung cancer tumorigenesis. miRNAs and pseudogenes are crucial gene regulators in normal cells as well as in lung cancer cells, and they have both oncogenic and tumor-suppressive roles in lung cancer tumorigenesis. In this study, we have determined the relationship between lung cancer related oncogenes and miRNAs along with pseudogenes in lung cancer, and the results indicate their potential as biological markers for diagnostic and therapeutic purposes.


[Vertical HIV-I-transmission. I. Risk and prevention in pregnancy].
Without anti-HIV treatment, mother to child HIV-I transmission occurs in 15-30% of HIV positive pregnancies. Transmission occurs mostly in the last trimester or at birth. The maternal virus load in the last trimester and around birth is strongly related to the risk of HIV transmission to the child. This risk can be reduced during pregnancy by anti-HIV treatment and in certain cases by performing a caesarean section. It is recommended to determine the plasma virus load several times during pregnancy. If the virus load is found to be high, measurement of plasma anti-HIV drug concentrations and anti-HIV drug resistance may prompt modification of the anti-HIV drug regimen with the objective of achieving maximal suppression of virus replication in the last trimester.


Molecular probes for malignant melanoma imaging.
Malignant melanoma represents a serious public health problem and is a deadly disease when it is diagnosed at late stage. Though (18)F-fluorodeoxyglucose ((18)F-FDG) positron emission tomography (PET) has been widely used clinically for melanoma imaging, other approaches to specifically identify, characterize, monitor and guide therapeutics for malignant melanoma are still needed. Consequently, many probes targeting general molecular events including metabolism, angiogenesis, hypoxia and apoptosis in melanoma have been successfully developed. Furthermore, probes targeting melanoma associated targets such as melanocortin receptor 1 (MC1R), melanin, etc. have undergone active investigation and have demonstrated high melanoma specificity. In this review, these molecular probes targeting diverse melanoma biomarkers have been summarized. Some of them may eventually contribute to the improvement of personalized management of malignant melanoma.


Exploring Supramolecular Assembly Space of Cationic 1,2,4-Selenodiazoles: Effect of the Substituent at the Carbon Atom and Anions.
Chalcogenodiazoles have been intensively studied in recent years in the context of their supramolecular chemistry. In contrast, the newly discovered cationic 1,2,4-selenodiazole supramolecular building blocks, which can be obtained via coupling between 2-pyridylselenyl halides and nitriles, are virtually unexplored. A significant advantage of the latter is their facile structural tunability via the variation of nitriles, which could allow a fine tuning of their self-assembly in the solid state. Here, we explore the influence of the substituent (which derives from the nitrile) and counterions on the supramolecular assembly of cationic 1,2,4-selenodiazoles via chalcogen bonding.


Duration of general anaesthetic exposure in early childhood and long-term language and cognitive ability.
The anaesthetic dose causing neurotoxicity in animals has been evaluated, but the relationship between duration of volatile anaesthetic (VA) exposure and neurodevelopment in children remains unclear. Data were obtained from the Western Australian Pregnancy Cohort (Raine) Study, with language (Clinical Evaluation of Language Fundamentals: Receptive [CELF-R] and Expressive [CELF-E] and Total [CELF-T]) and cognition (Coloured Progressive Matrices [CPM]) assessed at age 10 yr. Medical records were reviewed, and children divided into quartiles based on total VA exposure duration before age three yr. The association between test score and exposure duration quartile was evaluated using linear regression, adjusting for patient characteristics and comorbidity. Of 1622 children with available test scores, 148 had documented VA exposure and were split into the following quartiles: ≤25, >25 to ≤35, >35 to ≤60 and >60 min. Compared with unexposed children, CELF-T scores for children in the first and second quartiles did not differ, but those in the third and fourth quartiles had significantly lower scores ([3 rd quartile - Unexposed] -5.3; 95% confidence interval [CI], (-10.2 - -0.4), [4 th quartile - Unexposed] -6.2; 95% CI, (-11.6 - -0.9). CELF-E showed similar findings, but significant differences were not found in CELF-R or CPM for any quartile. Children with VA exposures ≤35 min did not differ from unexposed children, but those with exposures >35 min had lower total and expressive language scores. It remains unclear if this is a dose-response relationship, or if children requiring longer exposures for longer surgeries have other clinical reasons for lower scores.


Paederus fuscipes dermatitis: a report of nine cases observed in Italy and review of the literature.
Paederus sp. is a beetle belonging to Staphylinidae family and Coleoptera order. Its distribution is worldwide, especially in hot climates. Over 600 species of Paederus are known, approximately 50 are able to cause an irritant contact dermatitis. When the beetle is accidentally crushed on the skin, it releases pederin, a potent toxin with vesicating action. In Europe, only anecdotical cases of Paederus sp. dermatitis have been reported. Since 1993, we have observed approximately 25 patients with suspected Paederus sp. dermatitis. In 9 the clinical diagnosis was confirmed because Paederus fuscipes were found. The case list includes 6 males and 3 females, ages ranging from 6 to 53 years (mean age: 26.3 years). Six patients presented with one lesion and 3 with 2 lesions. Eyelids (3 patients), shoulders (3), neck (2), cheek (1), breast (1), back (1) and calf (1) were involved. All patients showed erythema, 2 oedema, 2 blisters, 1 vesicles, 1 pustules and 1 crusts. In the patient with pustules, bacteriological examinations were negative. A biopsy was carried out in 4 patients. In the early stages, spongiosis with exocytosis of neutrophils was observed. Vesicles with neutrophils and necrosis of the epidermis were typical of the advanced stage. Oedema of the dermis and a mixed infiltrate, with eosinophils, neutrophils and lymphocytes, in the papillary and upper reticular dermis, were also observed. Direct immunofluorescence was negative in all patients. Paederus sp. should also be considered in temperate climates as a possible aetiological agent in cases of acute dermatitis.


Myopathy as the persistently isolated symptomatology of primary autoimmune hypothyroidism.
Although disorders of thyroid function may cause a wide range of muscle disturbances, an overt myopathy has been rarely reported as an isolated clinical presentation of hypothyroidism. We observed 10 patients (5 males and 5 females) who had been referred to the department of neurology because of muscular fatigability, myalgia, cramps, or proximal weakness. Laboratory investigation showed that all patients had hypothyroidism due to Hashimoto's thyroiditis (atrophic variant in 9/10). Classic symptoms/signs of hypothyroidism such as lethargy, constipation, cold intolerance, myxedematous facies, and/or bradycardia were absent, as assessed independently by the three coauthoring thyroidologists. Muscular complaints improved greatly and then disappeared after substitutive levothyroxine treatment. Muscle biopsy revealed nonspecific changes. Nicotinamide adenine dinucleotide reductase (NADH-TR)-hyporeactive cores were present in two patients (10% and 90% of type 1 fibers). On electron microscopy, the core areas showed disorganized myofibrils, Z-band streaming, rod formation, and paucity of mitochondria and glycogen granules. Desmin intermediate filaments were overexpressed only in some cores. The similarity of the pattern of desmin expression between hypothyroid cores and target lesions of denervated fibers supports the hypothesis that, at least in some of our patients, myopathy was the result of an impaired nerve-mediated action of thyroid hormones on skeletal muscle. Our observations suggest that an isolated myopathy as the sole manifestation of hypothyroidism is not a rare event. We postulate that our cases may constitute a peculiar subgroup of Hashimoto's thyroiditis patients: (1) the strikingly abnormal F/M ratio of 1:1; (2) the relatively younger age; (3) the rarity of the goitrous variant; (4) the unusual finding of antithyroglobulin (Tg-Ab) > antithyroid peroxidase (TPO-Ab). Thorough evaluation of thyroid function is appropriate in patients with myopathy of uncertain origin.


Overcoming the influence of chronic pain on older patients' difficulty with recommended self-management activities.
Many older patients with common chronic conditions also experience chronic pain. We examined how chronic pain affects patients' difficulty with recommended self-management activities and the potential intervening role of self-efficacy (the level of confidence in one's own ability to perform a specific task). We obtained data from a cross-sectional nationwide survey of older patients, primarily older men, with chronic health conditions (N = 543). We defined chronic pain as pain present most of the time for 6 months or more during the past year. We assessed ability to follow self-management recommendations by asking respondents to rate their level of difficulty in performing three commonly recommended activities as suggested by their doctor. More than 60% of survey respondents reported chronic pain. Chronic pain was significantly associated with difficulty exercising regularly (odds ratio [OR] = 1.57, 95% confidence interval [CI] = 1.04-2.37) and taking prescribed medications (OR = 3.08, 95% CI = 1.10-8.59) but not with following a recommended eating plan (OR = 1.16, 95% CI = 0.76-1.76). However, when we took self-efficacy into account, chronic pain was no longer significantly associated with either exercise or taking medications. Chronic pain is a prevalent condition among older patients and is associated with greater reported difficulty performing certain essential self-management activities. Self-efficacy, however, plays an important intervening role. Specifically, higher self-efficacy negated or reduced the association between chronic pain and reported difficulty exercising and taking medications. Promoting self-efficacy among older adults with multiple chronic health problems is a promising strategy to improve their ability to follow self-management recommendations.


Scleral fixation of intraocular lens in eyes with history of open globe injury.
To report the results and complications of scleral fixation of intraocular lens (SF-IOL) in traumatized eyes of children with iris defects and inadequate capsular support with technique customized to the condition of the eye. Retrospective review of pediatric eyes with a history of open globe injury and SF-IOLs. All eyes had undergone pars plana vitrectomy and lensectomy after primary repair of the laceration. Site of scleral fixation and limbal incision were selected according to the site of traumatic scar, remaining capsular support, and keratometry. Ten patients with a mean age of 6.1 years were included. Mean duration of aphakia was 18.7 months. Five eyes had adequate capsular support for one haptic of the IOL. The only intraoperative complication was mild ciliary body hemorrhage. Mean follow-up was 11.8 months. Uncorrected visual acuity did not improve postoperatively in only one eye due to severe corneal astigmatism. Best-corrected visual acuity improved in 6 eyes. Mean postoperative sphere and cylinder were 1.8 and -3.05 diopters, respectively. The only postoperative complication was decentration of a sulcus-fixed haptic needing reoperation in one eye. SF-IOL is a viable option for correcting traumatic aphakia and can have good results if customized to the condition of the eye.


Inhibitory renorenal reflexes: a role for substance P or other capsaicin-sensitive neurons.
In anesthetized rats, we examined whether inhibitory renorenal reflex responses to renal pelvic mechanoreceptor (MR) and chemoreceptor (CR) stimulation were mediated by substance P (SP)-containing neurons. Capsaicin (0.5 ng to 5 micrograms) injected into the renal pelvis increased afferent renal nerve activity (ARNA) dose dependently, from 60 +/- 19 to 333 +/- 105%. For a given ARNA response, a 100-fold higher dose was required when capsaicin was injected into the renal interstitium compared with the renal pelvis. Renal pelvic administration of SP (25 ng) increased ipsilateral ARNA by 126 +/- 34% and contralateral urine flow rate and urinary sodium excretion by 21 +/- 4 and 28 +/- 7%, respectively, a response similar to that produced by renal MR and CR stimulation. Mean arterial pressure was unaffected. Ipsilateral renal denervation abolished the contralateral diuresis and natriuresis produced by SP. In rats treated with capsaicin (950 mg/kg subcutaneously over 1 wk) to deplete sensory neurons of SP, renal MR and CR stimulation failed to elicit a renorenal reflex response. The data suggest that the renorenal reflex responses to renal MR and CR stimulation are mediated at least, in part, by SP neurons or other sensory neurons susceptible to depletion by capsaicin.


Integrating machine learning techniques into robust data enrichment approach and its application to gene expression data.
The availability of enough samples for effective analysis and knowledge discovery has been a challenge in the research community, especially in the area of gene expression data analysis. Thus, the approaches being developed for data analysis have mostly suffered from the lack of enough data to train and test the constructed models. We argue that the process of sample generation could be successfully automated by employing some sophisticated machine learning techniques. An automated sample generation framework could successfully complement the actual sample generation from real cases. This argument is validated in this paper by describing a framework that integrates multiple models (perspectives) for sample generation. We illustrate its applicability for producing new gene expression data samples, a highly demanding area that has not received attention. The three perspectives employed in the process are based on models that are not closely related. The independence eliminates the bias of having the produced approach covering only certain characteristics of the domain and leading to samples skewed towards one direction. The first model is based on the Probabilistic Boolean Network (PBN) representation of the gene regulatory network underlying the given gene expression data. The second model integrates Hierarchical Markov Model (HIMM) and the third model employs a genetic algorithm in the process. Each model learns as much as possible characteristics of the domain being analysed and tries to incorporate the learned characteristics in generating new samples. In other words, the models base their analysis on domain knowledge implicitly present in the data itself. The developed framework has been extensively tested by checking how the new samples complement the original samples. The produced results are very promising in showing the effectiveness, usefulness and applicability of the proposed multi-model framework.


[Acute severe pancreatitis caused by anti-HIV drugs].
Acute pancreatitis in patients with HIV infection is infrequent and often not recognized. Autopsies performed on patients died for different pathologies show pancreatic lesions approximately in 50% of cases. The etiological factors associated with acute pancreatitis are related to HIV infection and particularly to drugs used to treat the syndrome or to prevent opportunistic infections. We report a case in a 38 years old male patient with AIDS without risk factors for acute pancreatitis in which the disease developed while taking Stavudine, Lamivudine and Indinavir. His conditions improved with intensive medical therapy. The subsequent medical course became complicated. The patient died after 75 days for severe abdominal haemorrhage. A review of the literature reports several cases of A.P. in HIV patients mainly related to opportunistic infections. Many authors regard the medical treatment of HIV syndrome as etiological factor but a casual relationship was never demonstrated. The aim of the present paper is to underline that besides Lamivudine also Stavudine and Indinavir might be related to acute pancreatitis. Clinical and laboratory investigations in HIV patients are therefore indicated for an earlier diagnosis and treatment.


Defect Engineering of ZnO Nanoparticles for Bioimaging Applications.
Many promising attributes of ZnO nanoparticles (nZnO) have led to their utilization in numerous electronic devices and biomedical technologies. nZnO fabrication methods can create a variety of intrinsic defects that modulate the properties of nZnO, which can be exploited for various purposes. Here we developed a new synthesis procedure that controls certain defects in pure nZnO that are theorized to contribute to the n-type conductivity of the material. Interestingly, this procedure created defects that reduced the nanoparticle band gap to ∼3.1 eV and generated strong emissions in the violet to blue region while minimizing the defects responsible for the more commonly observed broad green emissions. Several characterization techniques including thermogravimetric analysis, Fourier-transform infrared spectroscopy, X-ray photoelectron spectroscopy, transmission electron microscopy, Raman, photoluminescence, and inductively coupled plasma mass spectrometry were employed to verify the sample purity, assess how modifications in the synthesis procedure affect the various defects states, and understand how these alterations impact the physical properties. Since the band gap significantly decreased and a relatively narrow visible emissions band was created by these defects, we investigated utilizing these new nZnO for bioimaging applications using traditional fluorescent microscopy techniques. Although most nZnO generally require UV excitation sources to produce emissions, we demonstrate that reducing the band gap allows for a 405 nm laser to sufficiently excite the nanoparticles to detect their emissions during live-cell imaging experiments using a confocal microscope. This work lays the foundation for the use of these new nZnO in various bioimaging applications and enables researchers to investigate the interactions of pure nZnO with cells through fluorescence-based imaging techniques.


Searching HPV genome for methylation sites involved in molecular progression to cervical precancer.
Background: Human Papilloma Virus has been considered as the main cause for cervical cancer. In this study we investigated epigenetic changes and especially methylation of specific sites of HPV genome. The main goal was to correlate methylation status with histological grade as well as to determine its accuracy in predicting the disease severity by establishing optimum methylation cutoffs. Methods: In total, sections from 145 cases genotyped as HPV16 were obtained from formalin- fixed, paraffin-embedded tissue of cervical biopsies, conization or hysterectomy specimens. Highly accurate pyrosequencing of bisulfite converted DNA, was used to quantify the methylation percentages of UTR promoter, enhancer and 5' UTR, E6 CpGs 494, 502, 506 and E7 CpGs 765, 780, 790. The samples were separated in different groupings based on the histological outcome. Statistical analysis was performed by SAS 9.4 for Windows and methylation cutoffs were identified by MATLAB programming language. Results: The most important methylation sites were at the enhancer and especially UTR 7535 and 7553 sites. Specifically for CIN3+ (i.e. HSIL or SCC) discrimination, a balanced sensitivity vs. specificity (68.1%, 66.2% respectively) with positive predictive value (PPV) and negative predictive value (NPV) (66.2%, 68.2% respectively) was achieved for UTR 7535 methylation of 6.1% cutoff with overall accuracy 67.1%, while for UTR 7553 a sensitivity 60.9%, specificity 69.0%, PPV=65.6%, NPV=64.5% and overall accuracy=65.0% at threshold 10.1% was observed. Conclusion: Viral HPV16 genome was found methylated in NF-1 binding sites of UTR in cases with high grade disease. Methylation percentages of E6 and E7 CpG sites were elevated at the cancer group.


Development and application of purified tissue dissociation enzyme mixtures for human hepatocyte isolation.
Human hepatocyte transplantation is gaining acceptance for the treatment of liver diseases. However, the reagents used to isolate hepatocytes from liver tissue are not standardized and show lot-to-lot variability in enzyme activity and endotoxin contamination. For clinical application, highly purified reagents are preferable to crude digest preparations. A purified tissue dissociating enzyme (TDE) preparation (CIzyme(TM) purified enzymes) was developed based on the enzyme compositions found in a superior lot of collagenase previously used by our group for human hepatocyte isolation. The performance of this enzyme preparation was compared to collagenase type XI on 110 liver cases by assessing hepatocyte yield, viability, and seven other functional assays that included plating efficiency, basal and induced CYP450 activities, phase II conjugation activity, and ammonia metabolism. No statistically significant difference was observed between these TDEs when they were used to isolate hepatocytes from liver resections or organ donor tissue on 54 hepatocyte isolations with type XI enzyme and 56 isolations using CIzyme(TM). These results show that a highly purified and defined TDE preparation can be formulated that provides excellent performance with respect to viability, yield, and functional activity of the isolated cells. In addition to reproducible formulation, these purified enzyme products have only 2-3% of the endotoxin of crude enzyme preparations. These results show that purified enzymes such as CIzyme(TM) will be a safe and effective for the isolation of human hepatocytes for clinical transplants.


A lexicon for measuring maintenance of behavior change.
To establish a workgroup within the NIH-funded Health Maintenance Consortium (HMC) to examine how "maintenance" of behavior change was conceptualized and measured across and within behaviors. Multiple meetings were held by the workgroup to reach consensus definitions of maintenance and maintenance-related constructs across diet/nutrition, tobacco, substance abuse, and physical activity. Once consensus was reached, a survey assessed how maintenance was operationalized across 16 HMC intervention studies. Seventy-five percent of 16 studies assessed are using a criterion to assess maintenance and are tracking maintenance as a continuous measure. Eighty-one percent are assessing facilitators and barriers, and conceptualizing maintenance as both an intermediate and primary outcome measure. All 16 studies are assessing maintenance at the individual level with fewer at the organizational (N = 3), environmental (N = 3), and policy levels (N = 1). This survey found similarities and differences in measurement across behaviors that have important implications for advancing the quality of transbehavioral research.


Prostaglandin E2 activates outwardly rectifying Cl(-) channels via a cAMP-dependent pathway and reduces cell motility in rat osteoclasts.
We examined changes in electrical and morphological properties of rat osteoclasts in response to prostaglandin (PG)E(2). PGE(2) (>10 nM) stimulated an outwardly rectifying Cl(-) current in a concentration-dependent manner and caused a long-lasting depolarization of cell membrane. This PGE(2)-induced Cl(-) current was reversibly inhibited by 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS), 5-nitro-2-(3-phenylpropylamino)-benzoic acid (NPPB), and tamoxifen. The anion permeability sequence of this current was I(-) > Br(-) approximately Cl(-) > gluconate(-). When outwardly rectifying Cl(-) current was induced by hyposmotic extracellular solution, no further stimulatory effect of PGE(2) was seen. Forskolin and dibutyryl adenosine 3',5'-cyclic monophosphate (DBcAMP) mimicked the effect of PGE(2). The PGE(2)-induced Cl(-) current was inhibited by pretreatment with guanosine 5'-O-2-(thiodiphosphate) (GDPbetaS), Rp-adenosine 3',5'-cyclic monophosphorothioate (Rp-cAMPS), N-(2-[p-bromocinnamylamino]ethyl)-5-isoquinolinesulfonamide dihydrochloride (H-89), and protein kinase A inhibitors. Even in the absence of nonosteoclastic cells, PGE(2) (1 microM) reduced cell surface area and suppressed motility of osteoclasts, and these effects were abolished by Rp-cAMPS or H-89. PGE(2) is known to exert its effects through four subtypes of PGE receptors (EP1-EP4). EP2 and EP4 agonists (ONO-AE1-259 and ONO-AE1-329, respectively), but not EP1 and EP3 agonists (ONO-DI-004 and ONO-AE-248, respectively), mimicked the electrical and morphological actions of PGE(2) on osteoclasts. Our results show that PGE(2) stimulates rat osteoclast Cl(-) current by activation of a cAMP-dependent pathway through EP2 and, to a lesser degree, EP4 receptors and reduces osteoclast motility. This effect is likely to reduce bone resorption.


Comparison of Acute and Chronic Surgical Complications Following Robot-Assisted, Laparoscopic, and Traditional Open Radical Prostatectomy Among Men in Taiwan.
Few studies have evaluated long-term surgical complications in patients with prostate cancer (PC) who receive open radical prostatectomy (ORP), laparoscopic radical prostatectomy (LRP), or robot-assisted radical prostatectomy (RARP). To examine the perioperative and postoperative surgical complications among patients with PC who underwent ORP, LRP, or RARP. This cohort study included patients who received a diagnosis of resectable PC and underwent RP between January 1 and December 31, 2015. Participants were enrolled in the Taiwan Cancer Registry. The index date was the date of surgery, and the follow-up duration was the period from the index date to December 31, 2018. Data analysis was performed in September 2020. ORP, LRP, or RARP. Two multivariate mixed models accounting for hospital clusters were fitted to ascertain the association of RARP with treatment outcomes (ie, hospital stay, blood transfusion, postoperative pain, erectile dysfunction, urinary incontinence, and hernia); general linear regression models were used for continuous outcomes, the amount of blood transfused, and hospital stay, and logistic regression models were used for analyzing postoperative outcomes and surgical complications. Of the 1407 patients included in this study, 315 (22.4%) received ORP (mean [SD] age, 66.4 [6.8] years), 276 (19.6%) received LRP (mean [SD] age, 66.8 [6.4] years), and 816 (58.0%) received RARP (mean [SD] age, 66.1 [6.7] years). Mean (SD) follow-up in the full cohort was 36.7 (4.6) months. No statistically significant differences were observed in age, clinical tumor stage, pathological tumor stage, Gleason score, Gleason grade group, preoperative prostate-specific antigen concentration, D'Amico risk classification, and hospital level. A shorter hospital stay was observed for patients undergoing RARP vs those undergoing ORP (mean [SE] difference, -1.64 [0.22] days; P < .001) and LRP (mean [SE] difference, -0.57 [0.23] days; P = .01). Patients undergoing RARP had lower odds of receiving a blood transfusion (RARP vs ORP: adjusted odds ratio [aOR], 0.25; 95% CI, 0.17-0.36; RARP vs LRP: aOR, 0.58; 95% CI, 0.37-0.91). For postoperative pain, RARP was associated with a decrease in the odds of moderate to severe postoperative pain for as long as 12 weeks compared with both ORP and LRP (eg, RARP vs LRP at week 12: aOR, 0.40; 95% CI, 0.19-0.85; P = .02). The aORs for RARP vs those for ORP and LRP in the third year after RP were, for erectile dysfunction, 0.74 (95% CI, 0.45-0.92) and 0.60 (95% CI, 0.36-0.98), respectively; for urinary incontinence, 0.93 (95% CI, 0.65-0.99) and 0.60 (95% CI, 0.42-0.86), respectively; and for hernia, 0.51 (95% CI, 0.31-0.84) and 0.82 (95% CI, 0.46-0.92), respectively. In this study, undergoing RARP was associated with fewer acute and chronic postoperative complications than undergoing ORP or LRP.


Stabilizing Lead-Free All-Inorganic Tin Halide Perovskites by Ion Exchange.
Because of its thermal stability, lead-free composition, and nearly ideal optical and electronic properties, the orthorhombic CsSnI3 perovskite is considered promising as a light absorber for lead-free all-inorganic perovskite solar cells. However, the susceptibility of this three-dimensional perovskite toward oxidation in air has limited the development of solar cells based on this material. Here, we report the findings of a computational study which identifies promising Rb y Cs1-y Sn(Br x I1-x )3 perovskites for solar cell applications, prepared by substituting cations (Rb for Cs) and anions (Br for I) in CsSnI3. We show the evolution of the material electronic structure as well as its thermal and structural stabilities upon gradual substitution. Importantly, we demonstrate how the unwanted yellow phase can be suppressed by substituting Br for I in CsSn(Br x I1-x )3 with x ≥ 1/3. We predict that substitution of Rb for Cs results in a highly homogeneous solid solution and therefore an improved film quality and applicability in solar cell devices.


Gastric adenocarcinoma burden and late-stage diagnosis in Latino and non-Latino populations in the United States and Texas, during 2004-2016: A multilevel analysis.
Gastric cancer disproportionately affects Latinos, but little is known about regional effects and risk factors. We compared primary incidence, late-stage diagnosis, and risk factors for gastric adenocarcinoma (GCA) from 2004 to 2016 in Latinos and non-Latinos in the United States, Texas (TX), and South Texas (STX). We collected case data from Surveillance, Epidemiology, and End Results (SEER) and the Texas Cancer Registry. We generated average annual age-adjusted incidence rates, rate ratios (RRs), and 95% confidence intervals (CIs) using SEER*Stat software and analyzed the cases by anatomic site, demographics, and county-level risk factors using SAS 9.4. We constructed multilevel logistic regression models for late-stage GCA, adjusting for patient- and county-level characteristics. Latinos had higher overall GCA incidence rates in all regions, with the greatest disparities in overlap GCA in STX males (RR 4.39; 95% CI: 2.85, 6.93). There were no differences in cardia GCA rates for non-Hispanic Whites (NHWs) and Latino women in all regions. Younger patients, patients with overlapping or not otherwise specified (NOS) lesions, and patients diagnosed during 2012-2016 had higher odds of late-stage GCA. The stratification by location showed no differences in late-stage disease between NHWs and Latinos. The stratification by anatomic site showed Latinos with cardia GCA were more likely to have late-stage GCA than NHWs (OR: 1.13, p = 0.008). At the county level, higher odds of late-stage GCA were associated with medium and high social deprivation levels in TX without STX (OR: 1.25 and 1.20, p = 0.007 and 0.028, respectively), and medium social deprivation index (SDI) in patients with NOS GCA (OR: 1.21, p = 0.01). STX Latinos experience greater GCA disparities than those in TX and the United States. Younger age and social deprivation increase the risk for late-stage GCA, while Latinos and women are at higher risk specifically for late-stage cardia GCA. There is a need for population-specific, culturally responsive intervention and prevention measures, and additional research to elucidate contributing risk factors.


Erythropoietin directly stimulates osteoclast precursors and induces bone loss.
Erythropoietin (EPO) primarily regulates red blood cell formation, and EPO serum levels are increased on hypoxic stress (e.g., anemia and altitude). In addition to anemia, recent discoveries suggest new therapeutic indications for EPO, unrelated to erythropoiesis. We investigated the skeletal role of EPO using several models of overexpression (Tg6 mice) and EPO administration (intermittent/continuous, high/low doses) in adult C57Bl6 female mice. Using microcomputed tomography, histology, and serum markers, we found that EPO induced a 32%-61% trabecular bone loss caused by increased bone resorption (+60%-88% osteoclast number) and reduced bone formation rate (-19 to -74%; P < 0.05 throughout). EPO targeted the monocytic lineage by increasing the number of bone monocytes/macrophages, preosteoclasts, and mature osteoclasts. In contrast to the attenuated bone formation in vivo, EPO treatment in vitro did not inhibit osteoblast differentiation and activity, suggesting an indirect effect of EPO on osteoblasts. However, EPO had a direct effect on preosteoclasts by stimulating osteoclastogenesis in isolated cultures (+60%) via the Jak2 and PI3K pathways. In summary, our findings demonstrate that EPO negatively regulates bone mass and thus bears significant clinical implications for the potential management of patients with endogenously or therapeutically elevated EPO levels.


'I heard about this study on the radio': using community radio to strengthen Good Participatory Practice in HIV prevention trials.
During the Microbicides Development Program (MDP) 301, a clinical trial of a candidate microbicide amongst women in Johannesburg, South Africa, we used community radio to promote awareness of the trial, to inform community members about specific medical research procedures and terminologies, and to stimulate dialogue between researchers and local citizens. We used mixed methods to undertake a retrospective analysis of the social responses to the radio shows, focusing specifically on recruitment and participation in the MDP301 trial. We collected quantitative data that describes the themes and listener responses, the costs per broadcast, and the impact of the radio broadcasts on trial recruitment. Qualitative data on local reactions to the shows was gleaned from in-depth interviews with trial participants. Over a seven-year period, 205 individual broadcasts were made on two separate community radio stations. Show themes were either specifically related to medical research issues (36%), or focused on general health issues (46%), and sexual and reproductive health, including HIV prevention (18%). 403 listeners made telephone calls to the radio station, and 12% of women enrolled as participants in MDP301 (n = 9, 385) reported that they had first heard about the trial from the radio. Qualitative interviews (n = 401) with female MDP301 participants highlighted the effects of the radio shows in making women aware of the trial, impressing them with the importance of health screening and knowledge, legitimizing trial participation, and stimulating dialogue between trial participants and their male partners. Community radio is a potent tool for raising awareness and local knowledge about medical research and, in addition to other methodologies, can be used to promote recruitment into clinical trials. We suggest that future HIV prevention trials consider an investment in community radio beyond recruitment advertisements that incorporates this into the broader community engagement plan as a key element of Good Participatory Practice in clinical trial settings.


Infant Feeding Practices and Weight Gain in Toddlers Born Very Preterm: A Pilot Study.
Long-term consequences of prematurity are a public health concern. A pattern of slow initial weight gain followed by a period of rapid weight gain has been associated with poor cardiometabolic health outcomes. The purpose of this study was to examine the relationships between infant feeding practices and weight gain in a sample of 18-to-24-month olds corrected age born very preterm. A cross-sectional design was used to examine the relationships between infant feeding practices and weight gain. Estimates of effect sizes and model fit estimates were the primary parameters of interest. Most of the participants received human milk after birth, but most had transitioned to formula before three months. Slightly less than half received complementary foods prior to four months corrected age. Gains in weight and head circumference were rapid after discharge from the neonatal intensive care unit, while gains in length lagged behind. Infant feeding practices did not have a clinically meaningful effect on weight gain. While the initiation of human milk feedings was encouraging, the duration fell short of recommendations. Practices such as the early introduction of complementary feedings and the addition of rice cereal to the bottle are troubling. Additionally, the rapid increase in weight gain may have a negative impact on future cardiometabolic health. Clinical recommendations include ensuring support for the use of human milk before and after hospital discharge, close monitoring of physical growth, and ensuring adherence to the guidelines for the introduction of complementary foods.


Effects of Hypoxia and Bed Rest on Markers of Cardiometabolic Risk: Compensatory Changes in Circulating TRAIL and Glutathione Redox Capacity.
In chronic diseases, hypoxia and physical inactivity are associated with atherosclerosis progression. In contrast, a lower mortality from coronary artery disease and stroke is observed in healthy humans residing at high altitude in hypoxic environments. Eleven young, male volunteers completed the following 10-day campaigns in a randomized order: hypoxic ambulatory, hypoxic bed rest and normoxic bed rest. Before intervention, subjects were evaluated in normoxic ambulatory condition. Normobaric hypoxia was achieved in a hypoxic facility simulating 4000 m of altitude. Following hypoxia, either in bed rest or ambulatory condition, markers of cardiometabolic risk shifted toward a more atherogenic pattern consisting of: (a) lower levels of total HDL cholesterol and HDL2 sub-fraction and decreased hepatic lipase; (b) activation of systemic inflammation, as determined by C-reactive protein and serum amyloid A; (c) increased plasma homocysteine; (d) decreased delta-5 desaturase index in cell membrane fatty acids, a marker of insulin sensitivity. Bed rest and hypoxia additively decreased total HDL and delta-5 desaturase index. In parallel to the pro-atherogenic effects, hypoxia activated selected anti-atherogenic pathways, consisting of increased circulating TNF-related apoptosis-inducing ligand (TRAIL), a protective factor against atherosclerosis, membrane omega-3 index and erythrocyte glutathione availability. Hypoxia mediated changes in TRAIL concentrations and redox glutathione capacity (i.e., GSH/GSSG ratio) were greater in ambulatory conditions (+34 ± 6% and +87 ± 31%, respectively) than in bed rest (+17 ± 7% and +2 ± 27% respectively). Hypoxia-induced cardiometabolic risk is blunted by moderate level of physical activity as compared to bed rest. TRAIL and glutathione redox capacity may contribute to the positive interaction between physical activity and hypoxia. Highlights: - Hypoxia and bed rest activate metabolic and inflammatory markers of atherogenesis. - Hypoxia and physical activity activate selected anti-atherogenic pathways. - Hypoxia and physical activity positive interaction involves TRAIL and glutathione.


[Separation of fractions of phosphoprotein phosphatase and its protein inhibitors by isoelectrofocusing].
Individual molecular forms of phosphoprotein phosphatase from albino rat cardiac muscle were separated by isoelectrofocusing, resulting in a few fractions differing in pI (5.1, 5.4, 5.9-6.1 (double peak) and 7.1, respectively). Isoelectrofocusing of a purified enzyme preparation also allowed to isolate and characterize two protein inhibitors of the enzyme. The first one with pH 6.5-6.7 is similar to the thermostable phosphoprotein phosphatase inhibitor known from literature. This protein inhibits the enzyme activity by 90%; its effect is not decreased after 5-min heating at 95 degrees. The other inhibitor protein with pH 5.6-5.8 is thermolabile. When the enzyme activity was decreased 2.5-fold prior to thermal treatment, the latter protein lost this ability after heating at 95 degrees and inhibited the enzyme only by 9%. It is assumed that inhibitory proteins beside low molecular weight effectors can be involved in the mechanisms of the post-synthetical operative modification of phosphoprotein phosphatase function.


[Multi-sequence relation attention network for diagnosing HLA-B27-negative axial spondyloarthritis].
To develop a new 3D multi-sequence relation attention network for exploring the complementary and correlation information of different magnetic resonance imaging (MRI) modalities and improving the diagnostic performance of HLA-B27-negative axial spondyloarthropathy (axSpA). We retrospectively collected T1-weighted imaging (T1WI) and fat suppuration MRI (FS-MRI) data and clinical data of 375 HLA-B27-negative patients from the Third Affiliated Hospital of Southern Medical University (including 164 axSpA and 211 non-axSpA patients) and 49 patients from Nanhai Hospital (including 27 axSpA and 22 non-axSpA patients) between January, 2010 and August, 2021.A 3D relation attention network MSFANet based on multi-sequence MRI was used for automatic diagnosis of axSpA against non-axSpA in these patients.MSFANet consisted of a shallow shared feature learning module and a class-aware feature learning module, and latter module used a 3D multi-sequence relation attention mechanism to refine and fuse multi-sequence MRI features.A hybrid loss function was used to enhance the recognition ability of MSFANet by learning the loss weight coefficients of different branches to improve the classification performance. The experimental results demonstrated that MSFANet outperformed several state-of-the-art fusion algorithms (P<0.05) with AUC, accuracy, sensitivity, and specificity of 0.840, 77.93%, 83.70%, and 70.29% in the internal validation set, and of 0.783, 74.47%, 82.43% and 70.40% in the independent external validation set, respectively.The ablation studies showed that under the same architecture, the fusion model was superior to single-sequence models, which confirmed the effectiveness and necessity of fusing multi-sequence MRI.The visualization results demonstrated that MSFANet could focus on learning information from abnormal areas on MRI during the classification. We successfully constructed a 3D deep neural network based on multi-sequence MRI for differential diagnosis of HLA-B27-negative axSpA against nonaxSpA and verified the effectiveness of the multisequence relation attention mechanism for promoting classification performance of the network.


No H- and L-type cases in Belgium in cattle diagnosed with bovine spongiform encephalopathy (1999-2008) aging seven years and older.
The bovine spongiform encephalopathy (BSE) epidemic presented homogeneity of the phenotype. This classical BSE (called C-type) was probably due to the contamination of the food chain by a single prion strain. However, due to the active surveillance and better techniques, two rare variants of BSE have been recently reported in different continents without a clear correlation to the BSE epidemic. These emerging types behave as different strains of BSE and were named H-type and L-type according to the high and low molecular mass of the unglycosylated fragment of their proteinase K resistant prion protein (PrPres). In these types, the proportion of the un-, mono- and di-glycosylated fragments of PrP (glycoprofile) is also atypical and represents an effective diagnostic parameter. This study evaluated the presence of such types in bovine of 7 years and older in Belgium. The Belgian BSE archive contained 41 bovines of at least 7 years of age. The biochemical features of their PrPres were analyzed by Western blot with five antibodies recognising different regions of PrPres, from N- to C-terminus: 12B2, 9A2, Sha31, SAF84 and 94B4. All antibodies clearly detected PrPres except 12B2 antibody, which is specific for N-terminal region 101-105, a PrP region that is only retained in H-types. The glycoprofiles did correspond to that of C-type (with more than 55% of diglycosylated PrPres using antibody 94B4). Therefore, all cases have the features of C-type BSE. This study supports that, among the BSE cases of 7 years and older identified in Belgium, none was apparently of the H- or L- type. This is consistent with the very rare occurrence of atypical BSE and the restricted dimension of Belgium. These results shed some light on the worldwide prevalence of atypical BSE.


Urinary tract infection following ritual Jewish circumcision.
Circumcision seems to reduce the overall incidence of urinary tract infections (UTI), although a few studies have suggested that ritual circumcision may be a predisposing factor for UTI within the first 2 weeks following the procedure. The aim of this study was to investigate a possible causal relationship between ritual circumcision and UTI. The study comprised 82 infants with UTI, 55 females and 27 males under the age of 1 year. All males were circumcised on the eighth day of life. The median age of infection was 0.75 and 7.0 months for males and females, respectively. Fifty-two percent (14/27) of UTI episodes were diagnosed within the 2 weeks following circumcision. A significantly lower incidence in Escherichia coli-induced UTI was observed in males compared to females, 67% and 93%, respectively. Similarly, the incidence of E. coli-induced UTI was also significantly lower in males presenting within 2 weeks following circumcision (57%) compared to infants presenting prior or more than 2 weeks following the procedure (92%). Positive blood cultures of an identical microorganism were observed in 6/27 males compared to 2/55 females. The incidence of urinary tract malformations and their severity were similar in both sexes. We conclude that the high incidence of UTI following a ritual Jewish circumcision, as well as the relatively high preponderance of bacteria other than E. coli, may suggest a causal relationship between circumcision and UTI.


Temporal and spatial heterogeneity of mtDNA polymorphisms in natural populations of Drosophila mercatorum.
Restriction endonuclease analysis of mtDNA was used to examine the genetic relatedness of several geographically separated isolines of the Drosophila mercatorum subgroup. In addition, we examined the temporal and spatial distribution of two mtDNA restriction site polymorphisms produced by the enzymes BstEII and BstNI at a single locality--Kamuela, Hawaii. Due to small sample sizes of some collections and the undesirable dependance of the estimation of polymorphism frequency on its variance, an arcsin square root transformation of the frequency data was used. We also use an Fst estimator of our transformed frequencies to demonstrate considerable spatial and temporal differentiation within the Kamuela population. In contrast, isozyme data from the same population reveals no pattern of differentiation. The temporal and geographic heterogeneity and population subdivision detected with mtDNA analysis also is consistent with the known dispersal behavior and ecological constraints of this species. The mtDNA data in conjunction with the isozyme data show that the population structure of the Kamuela D. mercatorum is close to the boundary line separating panmixia from subdivision, a conclusion that could not be made from isozyme data alone.


The orphan nuclear receptor Rev-erbalpha recruits the N-CoR/histone deacetylase 3 corepressor to regulate the circadian Bmal1 gene.
Transcriptional regulation plays a fundamental role in controlling circadian oscillation of clock gene expression. The orphan nuclear receptor Rev-erbalpha has recently been implicated as a major regulator of the circadian clock. Expression of Bmal1, the master regulator of circadian rhythm in mammals, is negatively correlated with Rev-erbalpha mRNA level, but the molecular mechanism underlying this regulation is largely unknown. Here we show that Rev-erbalpha dramatically represses the basal activity of the mouse Bmal1 gene promoter via two monomeric binding sites, both of which are required for repression and are conserved between mouse and human. Rev-erbalpha directly binds to the mouse Bmal1 promoter and recruits the endogenous nuclear receptor corepressor (N-CoR)/histone deacetylase 3 (HDAC3) complex, in association with a decrease in histone acetylation. The endogenous N-CoR/HDAC3 complex is also associated with the endogenous Bmal1 promoter in human HepG2 liver cells, where a reduction in cellular HDAC3 level markedly increases the expression of Bmal1 mRNA. These data demonstrate a new function for the N-CoR/HDAC3 complex in regulating the expression of genes involved in circadian rhythm by functioning as corepressor for Rev-erbalpha.


Tobacco Rattle Virus as a Tool for Rapid Reverse-Genetics Screens and Analysis of Gene Function in Cannabis sativa L.
Medical cannabis (Cannabis sativa L.) is quickly becoming a central agricultural crop as its production has continued to increase globally. The recent release of the cannabis reference genomes provides key genetic information for the functional analysis of cannabis genes. Currently, however, the established tools for in vivo gene functional analysis in cannabis are very limited. In this study, we investigated the use of the tobacco rattle virus (TRV) as a possible tool for virus-induced gene silencing (VIGS) and virus-aided gene expression (VAGE). Using leaf photobleaching as a visual marker of PHYTOENE DESATURASE (PDS) silencing, we found that VIGS was largely restricted to the agro-infiltrated leaves. However, when agro-infiltration was performed under vacuum, VIGS increased dramatically, which resulted in intense PDS silencing and an increased photobleaching phenotype. The suitability of TRV as a vector for virus-aided gene expression (VAGE) was demonstrated by an analysis of DsRed fluorescence protein. Interestingly, a DsRed signal was also observed in glandular trichomes in TRV2-DsRed-infected plants, which suggests the possibility of trichome-related gene function analysis. These results indicate that TRV, despite its limited spread, is an attractive vector for rapid reverse-genetics screens and for the analysis of gene function in cannabis.


An integrated model for the effect of budesonide on ACTH and cortisol in healthy volunteers.
Budesonide, a glucocorticosteroid, is used as a first-line treatment for asthma. The aim of the study was to develop a PK/PD model for the effect of budesonide on ACTH and cortisol. The modelling data were generated by conducting a single-blind, randomized, placebo-controlled cross-over study. Ten healthy volunteers inhaled placebo (Placebo Turbohaler) and 1600 microg budesonide (Pulmicort Turbohaler), with a wash-out period of 7 days between treatments. Baseline concentrations of cortisol and ACTH were measured after placebo treatment and concentrations of cortisol, ACTH and budesonide were assessed after budesonide treatment. A one-compartment disposition model was used for budesonide disposition. Based on indirect response models, two types of models, distinguishing between production driven by a sum of cosine functions and production driven by surges, were used in parallel to describe the data. The surge-based approach was the most appropriate, based on goodness-of-fit, objective function values and number of parameters. The surge-based model that integrated both ACTH and cortisol data was chosen as the final model. The estimated half-lives of endogenous ACTH and cortisol were 9 and 113 min, respectively. The budesonide and ACTH concentrations producing 50% of the maximal response (IC(50) and A(50)) were 0.325 microg l(-1) and 4.96 pmol l(-1). The present PK/PD model of the effect of budesonide on ACTH and cortisol can serve as a tool for further understanding of the hypothalamic-pituitary-adrenal (HPA) axis and be useful in the development of drugs interacting with the axis.


Effect of method of collection on seminal characteristics of the domestic cat.
The effect of level of voltage and method of collection on seminal characteristics were studied in the domestic cat. A Latin square design was used to determine the effects of voltage (1, 2, 4, or 6 V) on seminal characteristics of electroejaculates for 8 cats (experiment 1). There was a significant effect of cat on the total volume (P less than 0.005), number of spermatozoa (P less than 0.05), pH (P less than 0.05), and osmolality (P less than 0.025). There was an effect of week for the pH (P less than 0.05) and osmolality (P less than 0.005) of semen. Voltage of stimulation affected the total volume (P less than 0.005), total number of spermatozoa (P less than 0.025), and osmolality (P less than 0.005) of semen. There were trends (P less than 0.1) for an effect of cat and an effect of voltage on spermatozoal motility. Urine contamination was not observed in any of the electroejaculates. A 2, 2 X 2 Latin square design was used in experiment 2 to determine the effect of method of collection (artificial vagina or electroejaculation) on seminal characteristics for 4 cats. Electroejaculation was performed, using the 6-V electrical stimulus selected from experiment 1. There was a significant (P less than 0.025) effect of cat on the motility of the spermatozoa in the viability preparation and a trend (P less than 0.1) for an effect of cat on spermatozoal viability.(ABSTRACT TRUNCATED AT 250 WORDS)


Actin genes in the Mediterranean fruit fly, Ceratitis capitata.
We have undertaken the study of actin gene organization and expression in the genome of the Mediterranean fruit fly (medfly), Ceratitis capitata. Actin genes have been extensively characterized previously in a wide range of eukaryotic organisms, and they have valuable properties for comparative studies. These genes are typically highly conserved in coding regions, represented in multiple copies per genome and regulated in expression during development. We have isolated a gene in the medfly using the cloned Drosophila melanogaster 5C actin gene as a probe. This medfly gene detects abundant messages present during late larval and late pupal development as well as in thoracic and leg tissue preparations from newly emerged adults. This pattern of expression is consistent with what has been seen for actin genes in other organisms. Using either the D. melanogaster 5C actin gene or the medfly gene as a probe identifies five common cross reacting EcoRI fragments in genomic DNA, but only under less than fully stringent hybridization conditions.


Development and validation of novel biomarker assays for osteoarthritis.
Osteoarthritis (OA) is the most common chronic joint disease usually diagnosed at relatively advanced stages when there is irreparable damage to the joint(s). Recently, we have identified two novel biomarkers C3f and V65 which appear to be OA-specific and therefore potential markers of early disease. We report the development of immunoassays for quantitative measure of these two novel biomarkers. Monoclonal and polyclonal antibodies were generated by immunising mouse and rabbits respectively with peptide-carrier conjugates of C3f and V65. Affinity purified antibodies were used for immunoassays development and assays validated using serum from OA patients and controls. The ELISAs developed showed spiked recovery of up to 96% for C3f and V65 peptides depending on serum dilutions with a coefficient of variation (CV) <10%. The intra- and inter-assay CVs for C3f and V65 were 1.3-10.8% and 4.2-10.3% respectively. Both assays were insensitive for measurements of the peptides in patients and the use of different signal amplification systems did not increase assay sensitivity. We have developed two immunoassays for measurements of C3f and V65 peptides biomarkers discovered by our earlier proteomic study. These assays could detect the endogenous peptides in serum samples from patients and controls but lacked sensitivity for accurate measurements of the peptides in patients. Our study highlights the difficulties and challenges of validating biomarker from proteomic studies and demonstrates how to overcome some of the technical challenges associated with developing immunoassays for small peptides.


Relationship between physical activity and type of smoking behavior among adolescents and young adults in Cyprus.
Research suggests that physical activity protects against youth smoking. As a result, prevention programs that focus on physical activity to address the problem of smoking hold promise. However, more research is needed to examine the relationship between physical activity and smoking and how physical activity might be used in smoking prevention and intervention programs. The present study examined the relationship between physical activity and smoking among adolescents and young adults using a sample from Cyprus. We investigated the association between level of physical activity (very active, active, moderately active, or inactive) and type of smoking behavior (nonsmoker; occasional, regular light, or regular heavy smoker). Participants were 1,390 Cypriot adolescents and young adults who completed a packet of questionnaires related to smoking and physical activity. Physical activity and demographic factors were analyzed in terms of their effect on smoking behavior. A consistent and negative relationship between physical activity and smoking emerged across both sex and age. The greater the intensity of youths' physical activity, the less likely they were to smoke. Compared with inactive individuals, physically active individuals smoked fewer cigarettes and were more likely to be nonsmokers or occasional smokers. Among physically active individuals, those engaging in more intense activity were less likely to be heavy or light smokers. Results of the present study indicate that physical activity may be used as a preventive measure against smoking in adolescents and young adults.


Chondrogenesis of mesenchymal stem cells in a novel hyaluronate-collagen-tricalcium phosphate scaffolds for knee repair.
Scaffolds are expected to play a key role in the induction of chondrogenesis of mesenchymal stem cells (MSCs) for cartilage tissue regeneration. Here, we report the development of a novel tricalcium phosphate-collagen-hyaluronate (TCP-COL-HA) scaffold that can function as a stem cell carrier to induce chondrogenesis and promote cartilage repair, and the investigation of chondroinductive properties of scaffolds containing varying amounts of TCP, COL and HA. TCP-COL-HA scaffolds, as well as TCP-COL scaffolds at two different TCP/COL ratios (50:50 and 25:75), were evaluated for their ability to induce cartilage regeneration from rabbit mesenchymal stem cells (rMSCs) in vitro and in vivo. Chondrogenic differentiation was evaluated by sulphated glycosaminoglycan quantification, collagen type II immunohistochemistry, and qRT-PCR. Mechanical strength was evaluated by the compression test. The results showed that the TCP-COL-HA scaffolds enhanced rMSC chondrogenesis to a greater degree than did the TCP-COL scaffolds; for the latter, the scaffold with the lower TCP/COL ratio (25:75) was superior in terms of promoting rMSC chondrogenesis. Similar results were obtained in an ectopic implantation model in nude mice. In a critical-size rabbit osteochondral defect-repair model, rMSCs seeded on TCP-COL-HA scaffolds showed greater cartilage regeneration and integration into surrounding tissue than the TCP-COL groups, in which cartilage repair was more efficient at the 25:75 than at the 50:50 ratio. These results indicate that the addition of HA and different TCP/COL ratios can affect the chondroinductive capacity of scaffolds, and suggest that the TCP-COL-HA scaffold can serve as an effective cell carrier for cartilage regeneration.


Neurotoxicity by synthetic androgen steroids: oxidative stress, apoptosis, and neuropathology: A review.
Anabolic-androgenic steroids (AAS) are synthetic substances derived from testosterone that are largely employed due to their trophic effect on muscle tissue of athletes at all levels. Since a great number of organs and systems are a target of AAS, their adverse effects are primarily on the following systems: reproductive, hepatic, musculoskeletal, endocrine, renal, immunological, infectious, cardiovascular, cerebrovascular, and hematological. Neuropsychiatric and behavioral effects as a result of AAS abuse are well known and described in the literature. Mounting evidence exists suggesting that in addition to psychiatric and behavioral effects, non-medical use of AAS carries neurodegenerative potential. Although, the nature of this association remains largely unexplored, recent animal studies have shown the recurrence of this AAS effect, ranging from neurotrophin unbalance to increased neuronal susceptibility to apoptotic stimuli. Experimental and animal studies strongly suggest that apoptotic mechanisms are at least in part involved in AAS-induced neurotoxicity. Furthermore, a great body of evidence is emerging suggesting that increased susceptibility to cellular oxidative stress could play a pivotal role in the pathogenesis of many neurodegenerative disorders and cognitive impairment. As in other drug-evoked encephalopathies, the key mechanisms involved in AAS - induced neuropathology could represent a target for future neuroprotective strategies. Progress in the understanding of these mechanisms will provide important insights into the complex pathophysiology of AAS-induced neurodegeneration, and will pave the way for forthcoming studies. Supplementary to abandoning the drug abuse that represents the first step in reducing the possibility of irreversible brain damage in AAS abusers, neuroprotective strategies have to be developed and implemented in future.


Escherichia coli capsule bacteriophages. IV. Free capsule depolymerase 29.
The free host capsule depolymerase, induced by Escherichia coli capsule bacteriophage no. 29, and causing the formation of haloes around its plaques, has been purified to homogeneity. As judged from the following facts, this "enzyme" consists of free phage 29 spikes. (i) Detached phage organelles and depolymerase 29 particles exhibit the same molecular weight (about 245,000, as determined from the sedimentation equilibrium), contain polypeptide chains of the same two sizes (57,000 plus or minus 3,000 and 29,500 plus or minus 2,000, as determined by SDS-PAA gel electrophoresis), and have (within experimental error) the same sedimentation coefficient, isoelectric point, and amino acid composition. (ii) Isolated depolymerase and phage spikes in situ both catalyze the hydrolysis of glucosidic bonds in host capsular polysaccharide, leading ultimately to the formation of oligosaccharide fragments of one, two, and three hexasaccharide repeating units. (iii) Depolymerase 29 and phage 29 spikes have roughly the same electron optical dimensions. As tentatively estimated from the total and the virus-associated capsule depolymerase activity in the lysates, phage 29 infection seems to produce eight to seventeen times more free than incorporated spikes.


β-blockers augment L-type Ca2+ channel activity by targeting spatially restricted β2AR signaling in neurons.
G protein-coupled receptors (GPCRs) transduce pleiotropic intracellular signals in mammalian cells. Here, we report neuronal excitability of β-blockers carvedilol and alprenolol at clinically relevant nanomolar concentrations. Carvedilol and alprenolol activate β2AR, which promote G protein signaling and cAMP/PKA activities without action of G protein receptor kinases (GRKs). The cAMP/PKA activities are restricted within the immediate vicinity of activated β2AR, leading to selectively enhance PKA-dependent phosphorylation and stimulation of endogenous L-type calcium channel (LTCC) but not AMPA receptor in rat hippocampal neurons. Moreover, we have engineered a mutant β2AR that lacks the catecholamine binding pocket. This mutant is preferentially activated by carvedilol but not the orthosteric agonist isoproterenol. Carvedilol activates the mutant β2AR in mouse hippocampal neurons augmenting LTCC activity through cAMP/PKA signaling. Together, our study identifies a mechanism by which β-blocker-dependent activation of GPCRs promotes spatially restricted cAMP/PKA signaling to selectively target membrane downstream effectors such as LTCC in neurons.


PPARγ Pro12Ala and ACE ID polymorphisms are associated with BMI and fat distribution, but not metabolic syndrome.
Metabolic Syndrome (MetS) results from the combined effect of environmental and genetic factors. We investigated the possible association of peroxisome proliferator-activated receptor-γ2 (PPARγ2) Pro12Ala and Angiotensin Converting Enzyme (ACE) I/D polymorphisms with MetS and interaction between these genetic variants. Three hundred sixty four unrelated Caucasian subjects were enrolled. Waist circumference, blood pressure, and body mass index (BMI) were recorded. Body composition was estimated by impedance analysis; MetS was diagnosed by the NCEP-ATPIII criteria. A fasting blood sample was obtained for glucose, insulin, lipid profile determination, and DNA isolation for genotyping. The prevalence of MetS did not differ across PPARγ2 or ACE polymorphisms. Carriers of PPARγ2 Ala allele had higher BMI and fat-mass but lower systolic blood pressure compared with Pro/Pro homozygotes. A significant PPARγ2 gene-gender interaction was observed in the modulation of BMI, fat mass, and blood pressure, with significant associations found in women only. A PPARγ2-ACE risk genotype combination for BMI and fat mass was found, with ACE DD/PPARγ2 Ala subjects having a higher BMI (p = 0.002) and Fat Mass (p = 0.002). Pro12Ala was independently associated with waist circumference independent of BMI and gender. Carriers of PPARγ2 Ala allele had higher BMI and fat-mass but not a worse metabolic profile, possibly because of a more favorable adipose tissue distribution. A gene interaction exists between Pro12Ala and ACE I/D on BMI and fat mass. Further studies are needed to assess the contribution of Pro12Ala polymorphism in adiposity distribution.


Effects of long-term estrogen implants in beef heifers.
Crossbred beef heifers (n = 78) were assigned randomly to one of three treatments. Heifers received either no implant, one estradiol-releasing implant (Compudose), or two estradiol-releasing implants. Heifers were implanted at birth and then reimplanted every 150 d. Calves were maintained with the cows until weaning at approximately 200 d of age. Heifers were placed in the feedlot as one group and fed a growing diet for 56 d. Following the growing phase, heifers were segregated into their respective treatment groups and fed until selected by industry buyers for harvest. Beginning at 1 yr of age and continuing every 14 d until puberty or harvest, heifers were palpated per rectum, and blood samples were collected for determination of ovarian activity and attainment of puberty. Serum progesterone of > or =1 ng/mL and(or) palpation of a detectable corpus luteum were criteria of puberty. At weaning and again at harvest, an x-ray was taken of the left front leg of six heifers selected randomly from each group. Metacarpal bones III and IV from the same animals were collected at harvest and transected for determination of epiphyseal plate closure. The x-ray scores and the actual measurements had a correlation of .94. Epiphyseal plate closure occurred in a dose-related manner, with heifers on the higher dose of estrogen having earlier plate closure than heifers on the lower dose. At harvest, reproductive tissues and carcass data were collected for all heifers. Eighteen of 25 untreated control heifers (P < .05), 6 of 26 heifers treated with one implant, and 2 of 26 heifers treated with two implants attained puberty by the end of the experiment. No differences (P > .10) were detected among treatment groups for carcass traits. These data suggest that early and continuous treatment of heifers with estradiol implants can retard reproductive function.


Leadership in applied psychology: Three waves of theory and research.
Although in the early years of the Journal leadership research was rare and focused primarily on traits differentiating leaders from nonleaders, subsequent to World War II the research area developed in 3 major waves of conceptual, empirical, and methodological advances: (a) behavioral and attitude research; (b) behavioral, social-cognitive, and contingency research; and (c) transformational, social exchange, team, and gender-related research. Our review of this work shows dramatic increases in sophistication from early research focusing on personnel issues associated with World War I to contemporary multilevel models and meta-analyses on teams, shared leadership, leader-member exchange, gender, ethical, abusive, charismatic, and transformational leadership. Yet, many of the themes that characterize contemporary leadership research were also present in earlier research. (PsycINFO Database Record


Slow light in multi-line Brillouin gain spectrum.
We present a method to achieve flat-top gain spectrum through overlapping multiple gain lines, which can be used to increase slow light bandwidth and relative pulse delay. A tunable gain bandwidth can be realized by changing the number of spectral lines and frequency separation between adjacent spectral lines. We demonstrate the method in a SBS-based slow light system. A phase modulator is used to modulate the phase of the pump wave, generating a pump wave with multi-line spectrum and achieving a Brillouin gain bandwidth of ~ 330 MHz.


The impact of patient weight on burn resuscitation.
Optimal fluid resuscitation of burn patients with burns greater than 20% total body surface area is critical to prevent burn shock during the initial 24 hours to 48 hours postburn. Currently, most resuscitation formulas incorporate the patient's weight when estimating 24-hour fluid requirements. The objective of this study was to determine the impact of weight on fluid resuscitation requirements and outcomes during the initial 24 hours after admission. We performed a retrospective review of patients admitted to our burn intensive care unit from December 2007 to April 2013, resuscitated with a computerized decision support system. We classified patients into body mass index (BMI) categories of underweight (BMI: <18.5), normal (BMI: 18.5-24.9), overweight (BMI: 25.0-29.9), or obese (BMI: >30.0). We also calculated the percent difference from ideal body weight (IBW) and compared 24-hour fluid volumes received. Patients with missing weight and/or height values were excluded from the study, resulting in a final cohort of 161 patients for analysis. Mean total body surface area was 42 ± 20% with a full thickness burn of 18 ± 23%. Mean age, weight, and height were 47 ± 19 years, 83 ± 19 kg, and 68 ± 4 inches, respectively. IBW for this cohort was 68 ± 11 kg with a BMI of 28 ± 6. Univariate analysis showed significant differences in 24-hour resuscitation volumes (mL/kg) between normal and obese patients (p < 0.05). Further analysis revealed that increasing percent difference from IBW was associated with lower fluid volumes. Although obesity was not associated with inhalation injury or renal replacement therapy, it was correlated to an increased risk for mortality (p < 0.05). This analysis showed that increasing weight was associated with lower fluid resuscitation volume requirements and a higher mortality rate, despite the low incidence of inhalation injury and renal replacement therapy in our obese patients. The use of actual body weight to drive resuscitation volumes may result in overresuscitation of obese patients, depending on the resuscitation formula. Further studies are needed to better explain the relationship between mortality and obesity in burn patients. Therapeutic/care management, level IV.


Bovine tuberculosis in Rwanda: Prevalence and economic impact evaluation by meat inspection at Société des Abattoirs de Nyabugogo-Nyabugogo Abattoir, Kigali.
Despite the significant public health burden of bovine tuberculosis (bTB) in Rwanda, the prevalence of bTB is poorly documented. This study was conducted to estimate the prevalence of bTB in cattle using gross examination of granulomatous lesions, to identify mycobacteria species in suspected samples, and to evaluate the economic impact of meat condemnation based on bTB-like lesions in the meat industry in Rwanda. Routine meat inspection was conducted at Société des Abattoirs de Nyabugogo (SABAN)-Nyabugogo Abattoir. Tissue samples including 31 lymph nodes, 3 lungs and 2 livers were obtained from cattle of different ages with gross tuberculous lesions. Mycobacterium bovis was identified using microscopy with Kinyoun staining and isolation of mycobacterial species in culture on Löwenstein-Jensen and Colestos media, further identified using biochemical tests. Our findings, based on culture and postmortem results, show that the prevalence of bTB is 0.5%(0.587*148/16753), with an overall gross tuberculous lesion prevalence of 0.9% (148/16753). The presence of lesions were higher in cattle aged 2 years and older (1.6% vs. 0.6%, p < 0.05) and higher in females than in males (1.4% vs. 0.6%, p < 0.05). Of the 36 samples tested, 26 (72.2%) were positive by microscopic examination with Kinyoun staining while M. bovis was culture-confirmed in 21 (58.7%) cases. Bovine tuberculosis caused condemnation of 1683.5 kg of meat, resulting in an estimated loss of $4810. Our findings indicate that the prevalence of bTB in Rwanda is significant, and that bTB is a major cause of meat condemnation requiring continued implementation of surveillance and control measures. Furthermore, the results from this study also show important variations in sensitivity of the different tests that were used to determine the prevalence of bTB in cattle in Rwanda.


Evolutionary genetics of Pasteurella haemolytica isolates recovered from cattle and sheep.
Genetic diversity and relationships among 194 Pasteurella haemolytica isolates, which were recovered predominantly from cattle (39%) and sheep (58%) suffering from pneumonic pasteurellosis in the United Kingdom, Germany, and the United States, were estimated by examination of allelic variation at 18 enzyme-encoding loci detected by multilocus enzyme electrophoresis. The isolates formed two major divisions. One included 178 Pasteurella haemolytica sensu stricto strains representing serotypes A1, A2, A5 to A9, A12 to A14, and A16; the other was composed of 16 isolates belonging to the A11 taxon. P. haemolytica isolates were classified into 22 electrophoretic types (ETs) that formed three primary phylogenetic lineages. One lineage was represented by ovine serotype A2 isolates, a second lineage consisted of bovine serotype A2, together with serotype A7 and A13 isolates, and the third lineage included isolates representing all of the other serotypes, as well as a second group of serotype A7 strains. Electrophoretic types were nonrandomly associated with specific capsular serotypes, lipopolysaccharide (LPS) types, outer membrane protein (OMP) types, and host species. Bovine isolates were represented by only three serotypes (A1, A2, and A6) in 5 ETs, whereas ovine isolates were represented by all of the serotypes in 19 ETs. The majority (76%) of bovine isolates were of serotypes A1 or A6 and belonged to a single ET that marked a virulent, cattle-specific clonal group. Among the ovine isolates, 40% were of serotype A2 and belonged to two ETs that represented two virulent, sheep-specific clonal groups. Bovine A1 and A6 isolates and bovine A2 isolates were phylogenetically distinct from ovine isolates of the same serotypes, indicating that different subpopulations of these serotypes are associated with disease in cattle and sheep. Consistent differences in the OMP profiles of strains of the bovine and ovine lineages of these three serotypes suggest that certain OMPs are involved in host specificity and virulence. Evolutionary relationships among P. haemolytica isolates indicate that the ancestral host is the sheep and that several distinct clonal lineages have crossed the species barrier into cattle. The A11 taxon is a heterogeneous group of opportunistic pathogens of sheep that represents a separate species.


A novel hypomorphic allele of Spag17 causes primary ciliary dyskinesia phenotypes in mice.
Primary ciliary dyskinesia (PCD) is a human condition of dysfunctional motile cilia characterized by recurrent lung infection, infertility, organ laterality defects and partially penetrant hydrocephalus. We recovered a mouse mutant from a forward genetic screen that developed many of the hallmark phenotypes of PCD. Whole-exome sequencing identified this primary ciliary dyskinesia only (Pcdo) allele to be a nonsense mutation (c.5236A>T) in the Spag17 coding sequence creating a premature stop codon (K1746*). The Pcdo variant abolished several isoforms of SPAG17 in the Pcdo mutant testis but not in the brain. Our data indicate differential requirements for SPAG17 in different types of motile cilia. SPAG17 is essential for proper development of the sperm flagellum and is required for either development or stability of the C1 microtubule structure within the central pair apparatus of the respiratory motile cilia, but not the brain ependymal cilia. We identified changes in ependymal ciliary beating frequency, but these did not appear to alter lateral ventricle cerebrospinal fluid flow. Aqueductal stenosis resulted in significantly slower and abnormally directed cerebrospinal fluid flow, and we suggest that this is the root cause of the hydrocephalus. The Spag17Pcdo homozygous mutant mice are generally viable to adulthood but have a significantly shortened lifespan, with chronic morbidity. Our data indicate that the c.5236A>T Pcdo variant is a hypomorphic allele of Spag17 that causes phenotypes related to motile, but not primary, cilia. Spag17Pcdo is a useful new model for elucidating the molecular mechanisms underlying central pair PCD pathogenesis in the mouse.This article has an associated First Person interview with the first author of the paper.


MicroRNA-200b and -301 are associated with gemcitabine response as biomarkers in pancreatic carcinoma cells.
Chemotherapy resistance (congenital or acquired) is one of the principal challenges for the treatment of pancreatic carcinoma. Recent evidence has demonstrated that epithelial to mesenchymal transition (EMT) is associated with chemoresistance in pancreatic carcinoma cells. However, the molecular mechanism underlying the development of chemoresistance remains unknown, and limited therapeutic options are available. Therefore, to anticipate individual chemosensitivity or acquired chemoresistance for patients with pancreatic carcinoma, predictive biomarkers are urgently required. Extensive evidence suggests that microRNAs (miRNAs) serve a crucial role in regulating EMT. The aim of this study was to examine the potential role of miRNA (miR)‑200b and miR‑301 in predicting the chemo‑responses to treatment for pancreatic carcinoma. The present results demonstrate that miR‑200b expression predicted chemo‑sensitivity and may have potential as a biomarker. In six different pancreatic carcinoma cell lines (Capan‑1, Capan‑2, Panc‑1, MIAPaCa‑2, BxPC‑3 and PL45 cells), the expression of miR‑200b correlated positively with chemosensitivity. Moreover, the enhanced expression of miR‑200b increased chemosensitivity and induced mesenchymal to epithelial transition. Conversely, miR‑301 modulated gemcitabine resistance and induced EMT through the downregulation of cadherin 1 expression. In addition, gemcitabine‑resistant cells (Capan‑2 and Panc‑1) exhibited upregulated miR‑301 expression and downregulated gemcitabine‑induced apoptosis. In summary, these two miRNAs may serve roles as biomarkers in pancreatic carcinoma, miR‑200b expression may predict chemosensitivity, and elevated miR‑301 expression may have potential applications in the prediction of acquired gemcitabine resistance.


How rapidly do self-compatible populations evolve selfing? Mating system estimation within recently evolved self-compatible populations of Azorean Tolpis succulenta (Asteraceae).
Genome-wide genotyping and Bayesian inference method (BORICE) were employed to estimate outcrossing rates and paternity in two small plant populations of Tolpis succulenta (Asteraceae) on Graciosa island in the Azores. These two known extant populations of T. succulenta on Graciosa have recently evolved self-compatibility. Despite the expectation that selfing would occur at an appreciable rate (self-incompatible populations of the same species show low but nonzero selfing), high outcrossing was found in progeny arrays from maternal plants in both populations. This is inconsistent with an immediate transition to high selfing following the breakdown of a genetic incompatibility system. This finding is surprising given the small population sizes and the recent colonization of an island from self-incompatible colonists of T. succulenta from another island in the Azores, and a potential paucity of pollinators, all factors selecting for selfing through reproductive assurance. The self-compatible lineage(s) likely have high inbreeding depression (ID) that effectively halts the evolution of increased selfing, but this remains to be determined. Like their progeny, all maternal plants in both populations are fully outbred, which is consistent with but not proof of high ID. High multiple paternity was found in both populations, which may be due in part to the abundant pollinators observed during the flowering season.


A population genetic comparison of argali sheep (Ovis ammon) in Mongolia using the ND5 gene of mitochondrial DNA; implications for conservation.
We sequenced 556 bp of the mitochondrial ND5 gene to infer aspects of population structure and to test subspecific designations of argali sheep (Ovis ammon) in Mongolia. Analysis of molecular variance (AMOVA) revealed greater variation within than among putative subspecies and populations, suggesting high levels female-mediated gene flow. Compared with bighorn sheep (O. canadensis) in North America, substantially less differentiation in mitochondrial DNA was found among argali populations over 1200 km than was found among bighorn populations over 250 km. This result is consistent with differences in argali and bighorn life history traits. Argali run for long distances across open terrain in the presence of a threat rather than running up into steep escape terrain like bighorn sheep do. Our results suggest recognizing only one Evolutionary Significant Unit (subspecies) of argali in Mongolia, but they may support recognizing two Management Units, because two regions do exhibit slightly different haplotype frequencies at the ND5 gene of mtDNA.


Low frequency of p53 gene mutation in tumors induced by aflatoxin B1 in nonhuman primates.
Aflatoxin B1 has been suggested as a causative agent for a G to T mutation at codon 249 in the p53 gene in human hepatocellular carcinomas from southern Africa and Qidong in China. To test this hypothesis, nine tumors induced by aflatoxin B1 in nonhuman primates were analyzed for mutations in the p53 gene. These included four hepatocellular carcinomas, two cholangiocarcinomas, a spindle cell carcinoma of the bile duct, a hemangioendothelial sarcoma of the liver, and an osteogenic sarcoma of the tibia. None of the tumors showed changes at the third position of codon 249 by cleavage analysis of the HaeIII enzyme site at codon 249. A point mutation was identified in one hepatocellular carcinoma at the second position of codon 175 (G to T transversion) by sequencing analysis of the four conserved domains (II to V) in the p53 gene. These data suggest that mutations in the p53 gene are not necessary in aflatoxin B1 induced hepatocarcinogenesis in nonhuman primates. The occurrence of mutation in codon 249 of the p53 gene in selective samples of human hepatocellular cancers may indicate involvement of environmental carcinogens other than aflatoxin B1 or that hepatitis B virus-related hepatitis is a prerequisite for aflatoxin B1 induction of G to T transversion in codon 249.


Tracing Sydenham's chorea: historical documents from a British paediatric hospital.
Sydenham's chorea (SC) became a well defined nosological entity only during the second half of the nineteenth century. Such progress was promoted by the availability of large clinical series provided by newly founded paediatric hospitals. This paper analyses the demographic and clinical features of patients with chorea admitted to the first British paediatric hospital (the Hospital for Sick Children, Great Ormond Street, London) between 1852 and 1936. The seasonal and demographic characteristics of SC during this time appear strikingly similar to those observed today, and witness the introduction of modern "statistically averaging" techniques in the approach to complex paediatric syndromes. Great Ormond Street (GOS) hospital case notes provide detailed descriptions of the "typical cases" of SC, and show that British physicians working in the early age of paediatric hospitals succeeded in recognising the most distinctive clinical features of this fascinating condition.


The effects of a calorie-reduced diet on periodontal inflammation and disease in a non-human primate model.
Low-calorie diets are commonplace for reducing body weight. However, no information is available on the effects of a reduced-calorie diet on periodontal inflammation and disease. The purpose of this study was to evaluate the clinical effects of a long-term calorie-restriction (CR) diet on periodontitis in an animal model of periodontitis. Periodontitis was induced in 55 young, healthy, adult rhesus monkeys (Macaca mulatta) by tying 2.0 silk ligatures at the gingival margins of maxillary premolar/molar teeth. Animals on a CR diet (30% CR; N = 23) were compared to ad libitum diet controls (N = 32). Clinical measures, including the plaque index (PI), probing depth (PD), clinical attachment level (CAL), modified gingival index (GI), and bleeding on probing (BOP) were recorded at baseline and 1, 2, and 3 months after ligature placement. Significant effects of CR were observed on the development of inflammation and the progression of periodontal destruction in this model. Compared to controls, CR resulted in a significant reduction in ligature-induced GI (P <0.0001), BOP (P <0.0015), PD (P <0.0016), and CAL (P <0.0038). Periodontal destruction, as measured by CAL, progressed significantly more slowly in the CR animals than in the controls (P <0.001). These clinical findings are consistent with available evidence that CR has anti-inflammatory effects. Moreover, these experimental findings are the first observations, to the best of our knowledge, that CR dampens the inflammatory response and reduces active periodontal breakdown associated with an acute microbial challenge.


Protocol for Past BP: a randomised controlled trial of different blood pressure targets for people with a history of stroke of transient ischaemic attack (TIA) in primary care.
Blood pressure (BP) lowering in people who have had a stroke or transient ischaemic attack (TIA) leads to reduced risk of further stroke. However, it is not clear what the target BP should be, since intensification of therapy may lead to additional adverse effects. PAST BP will determine whether more intensive BP targets can be achieved in a primary care setting, and whether more intensive therapy is associated with adverse effects on quality of life. This is a randomised controlled trial (RCT) in patients with a past history of stroke or TIA. Patients will be randomised to two groups and will either have their blood pressure (BP) lowered intensively to a target of 130 mmHg systolic, (or by 10 mmHg if the baseline systolic pressure is between 125 and 140 mmHg) compared to a standard group where the BP will be reduced to a target of 140 mmHg systolic. Patients will be managed by their practice at 1-3 month intervals depending on level of BP and followed-up by the research team at six monthly intervals for 12 months.610 patients will be recruited from approximately 50 general practices. The following exclusion criteria will be applied: systolic BP <125 mmHg at baseline, 3 or more anti-hypertensive agents, orthostatic hypotension, diabetes mellitus with microalbuminuria or other condition requiring a lower treatment target or terminal illness.The primary outcome will be change in systolic BP over twelve months. Secondary outcomes include quality of life, adverse events and cardiovascular events.In-depth interviews with 30 patients and 20 health care practitioners will be undertaken to investigate patient and healthcare professionals understanding and views of BP management. The results of this trial will inform whether intensive blood pressure targets can be achieved in people who have had a stroke or TIA in primary care, and help determine whether or not further research is required before recommending such targets for this population. ISRCTN29062286.


Assessing the clinical utility of biomarkers using the intervention probability curve (IPC).
Assessing the clinical utility of biomarkers is a critical step before clinical implementation. The reclassification of patients across clinically relevant subgroups is considered one of the best methods to estimate clinical utility. However, there are important limitations with this methodology. We recently proposed the intervention probability curve (IPC) which models the likelihood that a provider will choose an intervention as a continuous function of the probability, or risk, of disease. To assess the potential impact of a new biomarker for lung cancer using the IPC. The IPC derived from the National Lung Screening Trial was used to assess the potential clinical utility of a biomarker for suspected lung cancer. The summary statistics of the change in likelihood of intervention over the population can be interpreted as the expected clinical impact of the added biomarker. The IPC analysis of the novel biomarker estimated that 8% of the benign nodules could avoid an invasive procedure while the cancer nodules would largely remain unchanged (0.1%). We showed the benefits of this approach compared to traditional reclassification methods based on thresholds. The IPC methodology can be a valuable tool for assessing biomarkers prior to clinical implementation.


A rare case of hyaline-type Castleman disease in the liver.
Castleman disease often develops in the neck, mediastinum and pulmonary hilum. Its onset in the peritoneal cavity is very rare. The patient, a woman in her 70s, was referred to our department for a detailed examination of an abdominal mass. On abdominal ultrasonography, computed tomography scan, magnetic resonance imaging and positron emission tomography, a mass approximately 15 mm in diameter was noted in the hepatic S6. We attempted radical treatment and conducted a laparoscope-assisted right lobectomy. On the basis of histopathological findings, the patient was diagnosed as having hyaline type Castleman disease in the liver, a very rare condition.


Perindopril, converting enzyme blockade, and peripheral arterial hemodynamics in the healthy volunteer.
The effects of three doses (4, 8, and 16 mg) of perindopril, a new angiotensin I converting enzyme inhibitor, on systemic blood pressure, heart rate, brachial and carotid artery flow and diameter (assessed by the pulsed Doppler technique), forearm vascular resistance, plasma converting enzyme and renin activities, and plasma aldosterone were investigated in the normal volunteer and compared with those of a placebo over a 24-h period following oral drug intake in a double-blind, crossover trial. Perindopril dose-dependently decreased plasma converting enzyme activity, an effect that peaked at 3-4 h and persisted up to at least 48 h. Plasma renin activity increased for 12 h and plasma aldosterone was slightly decreased. Systemic blood pressure and heart rate were not drug-affected but perindopril dose-dependently augmented brachial and carotid artery flow, indicating an increase in peripheral arterial compliance. These vasodilating effects, which lasted up to 10 h after drug intake, affected both large arteries and arterioles, the latter being more sensitive, however, and were more marked in the muscular resistance vessels.