Skeletal Fluorosis: A Case of Inhalant Abuse Leading to a Diagnosis of Colon Cancer.
Skeletal fluorosis is a long-term bone disease that develops when prolonged fluoride toxicity leads to osteosclerosis and bone deformities that result in crippling pain and debility. The disease is endemic to many countries due to environmental or industrial exposures. However, rare cases in the United States have been reported from various causes including heavy toothpaste ingestion, excessive tea consumption, voriconazole use, and inhalant abuse. Here, we present a case of a 41-year-old man who presented for weight loss and severe joint pains due to bony sclerotic lesions found on X-rays. Social history revealed that he had been recreationally inhaling compressed air dusters used for cleaning electronics. Owing to concern for malignancy, he underwent an extensive work-up which led to a diagnosis of colon cancer, but positron emission tomography/computed tomography (PET/CT) and bone biopsy were unexpectedly negative for metastatic bone disease. Further characterization of his lesions by skeletal survey led to a diagnosis of skeletal fluorosis secondary to inhalant abuse. As in this patient, the disease can be difficult for clinicians to recognize as it can be mistaken for various boney diseases such as metastatic cancer. However, once there is clinical suspicion for skeletal fluorosis, various tests to help confirm the diagnosis can include serum and urine fluoride levels, skeletal survey, and bone ash fluoride concentration. Treatment of skeletal fluorosis primarily involves cessation of fluoride exposure, and recovery can take years. Ultimately, further study is required to develop recommendations and guidelines for diagnosis, management, and prognosis of the disease in the United States.


QM/MM calculations suggest a novel intermediate following the proton abstraction catalyzed by thymidylate synthase.
The cleavage of covalent C-H bonds is one of the most energetically demanding, yet biologically essential, chemical transformations. Two C-H bond cleavages are involved in the reaction catalyzed by thymidylate synthase (TSase), which provides the sole de novo source of thymidylate (i.e., the DNA base T) for most organisms. Our QM/MM free energy calculations show that the C-H → O proton transfer has three transition states that are energetically similar but structurally diverse. These characteristics are different from our previous calculation results on the C-H → C hydride transfer, providing an explanation for differences in temperature dependences of KIEs on these two C-H bond activation steps. The calculations also suggest that the traditionally proposed covalent bond between the protein and substrate (the C6-S bond) is very labile during the multistep catalytic reaction. Collective protein motions not only assist cleavage of the C6-S bond to stabilize the transition state of the proton transfer step but also rearrange the H-bond network at the end of this step to prepare the active site for subsequent chemical steps. These computational results illustrate functionalities of specific protein residues that reconcile many previous experimental observations and provide guidance for future experiments to examine the proposed mechanisms. The synchronized conformational changes in the protein and ligands observed in our simulations demonstrate participation of protein motions in the reaction coordinate of enzymatic reactions. Our computational findings suggest the existence of new reaction intermediates not covalently bound to TSase, which may lead to a new class of drugs targeting DNA biosynthesis.


Preoperative platelet lymphocyte ratio as independent predictors of prognosis in pancreatic cancer: A systematic review and meta-analysis.
Recently, the preoperative platelet to lymphocyte ratio (PLR) has been found reported to predict oncologic outcomes in multiple malignancies. However, its prognostic value in patients with pancreatic cancer (PC) remains controversial. The aim of this study was to assess the prognostic value of preoperative PLR in PC. MEDLINE, EMBASE, and Cochrane databases were searched to identify studies evaluating the prognostic significance of preoperative PLR in PC. Pooled hazard ratios (HRs) for overall survival (OS) were calculated using fixed-effects/random-effects models. A total of eight studies comprising 1,904 patients with PC were included in the meta-analysis. The pooled analysis demonstrated that elevated PLR had an association with decreased OS (HR: 1.22, 95% CI: 1.04-1.43, p = 0.02). Subgroup analysis showed that a high PLR significantly predicted poor OS in Asian studies (HR: 1.25, 95% CI: 1.03-1.52, p = 0.02), patients with metastatic disease (HR: 1.34, 95% CI: 1.01-1.77, p = 0.04) and patients with PLR >150 (HR: 1.73, 95% CI: 1.21-2.49, p = 0.003). The preoperative PLR may be a significant independent prognostic factor in patients with PC.


[Spectral-analytical studies of the action of etifoxin on the human EEG].
Spectral analysis of EEG in healthy volunteers shows that Etifoxin, the hydrochloride of 6-chloro-4-methyl-4-phenyl-2-ethylamino-4H-3,1-benzoxazine (Hoe 36,801), in an oral dosage of 100 mg has a differential effect in function of the individual alpha organization. This differential effect appearing in the course of the quantitative evaluation can be interpreted, from a morphological point of view, as an expression of various partial phases and patterns of a process leading to reduced vigilance.


A prospective study of PTSD and early-age heart disease mortality among Vietnam veterans: implications for surveillance and prevention.
To examine prospectively early-age heart disease (HD) among a national random sample of 4328 male Vietnam veterans, who did not have HD at baseline in 1985. Studies have suggested that posttraumatic stress disorder (PTSD) may result in cardiovascular disease. However, many past studies had important methodological limitations to their designs. Using Cox regressions, we assessed PTSD, age, race, intelligence, family history, obesity, smoking, alcohol abuse, antisocial personality, and depression in predicting HD mortality at follow-up in December 31, 2000. The men were <65 years old at follow-up. Using two PTSD measures, a Diagnostic and Statistical Manual of Mental Disorders, 3rd Edition (DSM-III) measure (D-PTSD) and one developed by Keane (K-PTSD), we found that among Vietnam theater and era veterans combined (era veterans had no Vietnam service), having PTSD was associated with HD mortality for D-PTSD (hazard ratio (HR) = 2.25, p = .045) and approached significance for K-PTSD (HR = 2.16, p = .066). However, having higher PTSD symptoms on either scale was associated with mortality, with a 5-point increase associated with approximately 20% increase in mortality risk (all p < .05). Controlling for lifetime depression only slightly altered the results. The effects for theater veterans alone were stronger (D-PTSD: HR = 2.58, p = .025; K-PTSD: HR = 2.73, p = .022). Among theater veterans, controlling for lifetime depression or combat exposure made little difference. PTSD was prospectively associated with HD mortality among veterans free of HD at baseline. This study suggests that early-age HD may be an outcome after military service among PTSD-positive veterans.


[Osteomyelitis, therapy and prognosis (author's transl)].
The application of antibiotics at the treatment of osteomyelitis resulted in a rise of the permanent successful treatment rate. This is especially true for the acute hematogenous osteomyelitis, to a certain extent also for the chronic exogenous one, at which even today the antibiotical therapy is not as important as a surgical intervention. Under consideration of the generally accepted therapeutical principles the special importance of a much individualized treatment adjusted to the clinical picture and disease stage is pointed out. The advantages and disadvantages of an application of antibiotics at the osteomyelitis are mentioned and we try to explain the reasons for occasional therapy failures. A special chapter is devoted to the methods of filling and sterilization of the osteomyelitis bone hollows. Our own experiences with an operative combination process are reported.


A refined method for molecular typing reveals that co-occurrence of PrP(Sc) types in Creutzfeldt-Jakob disease is not the rule.
Molecular typing in Creutzfeldt-Jakob disease (CJD) relies on the detection of distinct protease-resistant prion protein (PrP(Sc)) core fragments, which differ in molecular mass or glycoform ratio. However, the definition and correct identification of CJD cases with a co-occurrence of PrP(Sc) types remains a challenge. With antibodies recognizing a linear epitope in the octapeptide repeat PrP region, supposed to distinguish between the two major PrP(Sc) isoforms (ie, types 1 and 2), it was recently shown that all type 2 cases display an associated band with a type 1 migration pattern, which led to the conclusion that multiple PrP(Sc) types regularly coexist in CJD. We studied brain samples from 53 sporadic CJD and 4 variant CJD subjects using a high-resolution electrophoresis, a wide range of proteinase K (PK) activities, the 'type 1-selective' antibody 12B2, and several unselective antibodies. We found that the type 1-like band detected by 12B2 in all CJD subtypes associated with PrP(Sc) type 2 is not a PK-resistant PrP(Sc) core but rather matches the physicochemical properties of partially cleaved fragments, which result from the several PK cleavage sites included in the N-terminal portion of PrP(Sc). Furthermore, using gels with high resolution and a relatively high PK activity, we were able to increase the detection sensitivity of either type 1 or 2, when coexisting, to amount corresponding to 3-5% of the total PrP(Sc) signal (ie, weak band of one type/total PrP(Sc)). Our results show that there are many pitfalls associated with the use of 'type 1 selective' antibodies for CJD typing studies and that co-occurrence of PrP(Sc) types in CJD is not the rule. The present results further validate the rationale basis of current CJD classification and the qualitative nature of molecular typing in CJD.


Effects of nitric oxide synthase inhibitor on intraocular pressure and ocular inflammation following laser irradiation in rabbits.
To examine the effects of nitric oxide synthase (NOS) inhibitors on intraocular pressure (IOP) and ocular inflammation following laser irradiation of the rabbit iris, and to investigate the involvement of nitric oxide (NO). Thirty min after the intravenous administration of a nonselective inhibitor of NOS, N omega-nitro-L-arginine methyl ester (L-NAME, 1-100 mg/kg), or a selective inhibitor of iNOS, aminoguanidine (AG, 100 mg/kg), Q-switched Nd:YAG laser irradiation was applied to the iris of albino rabbits at an energy level of 48 mJ. IOP was measured prior to and for 24 h after irradiation. In separate groups of rabbits, aqueous humor was withdrawn 30 min after irradiation to determine protein and prostaglandin (PG) E2 concentrations. Intravenously administered L-NAME dose-dependently inhibited the acute increase in IOP, the peak of which was observed at 30 min, following laser irradiation. The IOP increase was completely abolished by 100 mg/kg of L-NAME. This dose of L-NAME significantly reduced the elevation of protein concentration in aqueous humor following irradiation; however, this dose failed to affect the increase in PGE2 concentration On the other hand, the inhibitory effects of AG (100 mg/kg) on the increase in IOP and aqueous protein following laser irradiation were not significant. Intravenous administration of L-NAME significantly inhibits the IOP rise and the increase in protein concentration in aqueous humor following laser irradiation, but AG does not, suggest the involvement of cNOS in these ocular responses to laser irradiation.


Pragmatic and idiosyncratic acts in human everyday routines: the counterpart of compulsive rituals.
Our daily activities are comprised of motor routines, which are behavioral templates with specific goals, typically performed in an automatic fixed manner and without much conscious attention. Such routines can seem to resemble pathologic rituals that dominate the motor behavior of obsessive-compulsive disorder (OCD) and autistic patients. This resemblance raises the question of what differentiates and what is common in normal and pathologic motor behavior. Indeed, pathologic motor performance is often construed as an extended stereotyped version of normal everyday routines. In this study we applied ethological tools to analyze six motor routines performed by 60 adult human volunteers. We found that longer normal everyday routines included more repetitions, but not more types of acts, and that in each routine, most acts were performed either by all individuals (pragmatic acts) or by only one individual (idiosyncratic components). Thus, normal routines consist in a relatively rigid part that is shared by all individuals that perform the routine, and a flexible part that varies among individuals. The present results, however, do not answer the question of whether the flexible individual part changes or remains constant over routine repetition by the same person. Comparing normal routines with OCD rituals revealed that the latter comprise an exaggeration of the idiosyncratic component. Altogether, the present study supports the view that everyday normal routines and pathologic rituals are opposite processes, although they both comprise rigid motor behavioral sequences.


Low Probability of Lymphatic Drainage to Cloquet's Node Is of Limited Value as Indicator for Pelvic Lymph Node Dissection in Patients with Lower Limb Melanoma.
For patients with melanoma, the choice between an inguinal lymph node dissection (ILND) alone and both an ILND and a pelvic lymph node dissection (PLND) is controversial. Although Cloquet's node (CN) is considered the sentinel pelvic node, evaluation of this factor to predict pelvic node status has produced varied results. We investigated inguinal and pelvic lymphatic drainage patterns and focused on CN to clarify whether CN status could be an indicator of PLND. Patients with primary cutaneous lower limb melanoma who underwent lymphatic mapping and sentinel lymph node biopsy (SLNB) using dynamic lymphoscintigraphy and SPECT/CT were retrospectively reviewed. Thirty-two patients underwent lymphatic mapping and SLNB. Each patient's CN was identified by SPECT/CT. A radioactive CN was detected in only 37.5% (12/32) of patients, and no lymphatic drainage to CN occurred in 62.5% (20/32). In 37.5% (12/32) of patients, the lymph drained directly from the inguinal to the pelvic nodes bypassing CN. In melanoma patients, lymphatic drainage from the lower extremity does not always pass from the inguinal node to the pelvic nodes via CN. Tumor-negative status of CN alone is of limited value as an indicator for avoiding PLND.


A novel technique for observing the internal ultrastructure of human chromosomes with known karyotype.
Observation of the internal ultrastructure of human chromosomes by transmission electron microscopy (TEM) has frequently been attempted in spite of the difficulties in detaching metaphase chromosome spreads from the glass slide for further processing. In this study we have used a method in which metaphase chromosome spreads were prepared on a flexible thermoplastic membrane (ACLAR) film. To assess chromosome identity, a diamidino-phenylindole staining and karyotying was first done using a conventional cytogenetic system. The chromosome spreads were then fixed with 1% osmium tetroxide, stained with freshly prepared 2% tannic acid, dehydrated, and flat-embedded in epoxy resin. The resin sheet was easily detachable and carried whole chromosome spreads. By this method, TEM observation of chromosomes from normal human lymphocytes allowed a thorough examination of the ultrastructure of centromeres, telomeres, fragile sites, and other chromosomal regions. Various ultrastructural patterns including thick electron dense boundaries, less dense internal regions, and extended chromatin loops at the periphery of the chromosomes were discernible. Application of the present method to chromosome research is expected to provide comprehensive information on the internal ultrastructure of different chromosomal regions in relation to function.


Pharmacophore Mapping, In Silico Screening and Molecular Docking to Identify Selective Trypanosoma brucei Pteridine Reductase Inhibitors.
Trypanosoma brucei Pteridine reductase (TbPTR1) is of vital importance and is an established drug target for dreaded Human African trypanosomiasis (HAT). Pharmacophore perception strategy has been employed to identify key chemical features responsible for the biological activity for TbPTR1. The findings suggest that three different pharmacophore features can be associated with T. brucei anti-PTR1 activity namely: H-bond donors (D), Hydrophobic aromatic (H) and Ring aromatic (R). The resulting hypothesis is able to predict the activity of other existing TbPTR1 inhibitors with a correlation coefficient (r) of 0.89. An in silico database screening, based on the best hypothesis, has been used to identify some potential nanomolar range TbPTR1 inhibitors. These compounds were then checked by molecular docking and subjected to ADMET analysis. Further, a detailed comparison of the pharmacophore behavior and differential analysis of binding pockets of T. brucei and L. major was made which revealed subtle differences in terms of their shape and charge properties. This investigation can form the basis for tweaking the specificity of compounds for generating new improved species specific inhibitor molecules for Pteridine reductase in these different parasitic protozoans.


Withaferin A induces mitochondrial-dependent apoptosis in non-small cell lung cancer cells via generation of reactive oxygen species.
Withaferin A (WA) is a bioactive lactone, isolated from natural sources, mainly found in Withania somnifera, and was known to be highly effective against a variety of tumor cells both in vitro and in vivo. Accumulating experimental evidence suggests the involvement of reactive oxygen species (ROS) in WA-mediated cytotoxicity against cancer cells. Hence, the purpose of this study was to investigate the effect of WA in non-small cell lung cancer (NSCLC) cells and also the role of ROC in WA-mediated cytotoxicity. In the present study we investigated the cytotoxic potential of WA against NSCLC cell line A549 and also highlighted the mechanism of cytotoxicity of this compound. Non-carcinoma WI-38 and PBMC cell lines were used as controls. WA treatment resulted in a dose-dependent cytotoxicity in A549 cells, while the non-carcinoma cells WI-38 and PBMC were unaffected. Further experimental approaches revealed that ROS plays a major role in WAinduced apoptosis in NSCLC cells. WA induces oxidative damage to NSCLC cells with minimum toxicity to normal cells.


Effect of slow versus fast desaturation on the ventilatory response to hypoxia.
The ventilatory response to progressive hypoxia was studied in 15 healthy adult males by employing two different rates of desaturation. In a slow hypoxic ramp, peripheral oxygen saturation was gradually reduced to 70% over 10-13 min, while in a fast ramp the saturation was reduced over 3-5 min. The ventilatory response to hypoxia was evaluated as the slope of the regression between oxygen desaturation and minute ventilation (l/min/%). There was no significant difference in the slope between the slow and fast ramps. The subjects were further divided into two subgroups; namely high and low responders, according to the value of their slopes. A slope of 0.6 l/min/% was considered the cut-off limit for this classification. In the group of high responders, the hypoxic ventilatory response was significantly higher with the slow ramp than with the fast ramp (p < 0.05). In the low responders, the response was significantly higher with the fast ramp (p < 0.05). We conclude that the hypoxic ventilatory responses are influenced by the rate of desaturation and they appear to be related to the individual sensitivity to hypoxia. These responses are reversed in the low compared with the high responders.


Initial binding and recellularization of decellularized mouse lung scaffolds with bone marrow-derived mesenchymal stromal cells.
Recellularization of whole decellularized lung scaffolds provides a novel approach for generating functional lung tissue ex vivo for subsequent clinical transplantation. To explore the potential utility of stem and progenitor cells in this model, we investigated recellularization of decellularized whole mouse lungs after intratracheal inoculation of bone marrow-derived mesenchymal stromal cells (MSCs). The decellularized lungs maintained structural features of native lungs, including intact vasculature, ability to undergo ventilation, and an extracellular matrix (ECM) scaffold consisting primarily of collagens I and IV, laminin, and fibronectin. However, even in the absence of intact cells or nuclei, a number of cell-associated (non-ECM) proteins were detected using mass spectroscopy, western blots, and immunohistochemistry. MSCs initially homed and engrafted to regions enriched in types I and IV collagen, laminin, and fibronectin, and subsequently proliferated and migrated toward regions enriched in types I and IV collagen and laminin but not provisional matrix (fibronectin). MSCs cultured for up to 1 month in either basal MSC medium or in a small airways growth media (SAGM) localized in both parenchymal and airway regions and demonstrated several different morphologies. However, while MSCs cultured in basal medium increased in number, MSCs cultured in SAGM decreased in number over 1 month. Under both media conditions, the MSCs predominantly expressed genes consistent with mesenchymal and osteoblast phenotype. Despite a transient expression of the lung precursor TTF-1, no other airway or alveolar genes or vascular genes were expressed. These studies highlight the power of whole decellularized lung scaffolds to study functional recellularization with MSCs and other cells.


[Optimization of combined brachy-teletherapy by the modification technic].
A method is presented for combined brachy-teletherapy by which the dose distribution of percutaneous irradiation is adapted to the distribution of short-distance irradiation with 192iridium by means of the so-called "field integrated dose modification". Insufficient or excessive doses within the area of superposition of both therapy modalities can so be avoided. Moreover, the therapist can define zones of deviation from the reference dose, if such a dose heterogeneity seems desirable in the individual case. The combined irradiation of prostata and cervix is given as an example to describe in detail the technical, clinical, and radiobiological aspects of this method.


The Correlation of SCL-90-R Anxiety, Depression, Somatization Subscale Scores with Chronic Low Back Pain.
To investigate the correlation of the Symptom Checklist-90-R (SCL-90-R) anxiety, depression, and somatization subscale scores with chronic low back pain (LBP). Seventy-five patients who were admitted with the complaint of chronic LBP (patient group) and 75 healthy persons (control group) were evaluated. SCL-90-R anxiety, depression, and somatization subscale scores of patients having chronic LBP and healthy persons were measured. The mean values were paired and using two tailed t test they were statistically evaluated. The difference between SCL-90-R anxiety subscale subscores of patients having chronic LBP and healthy persons was not statistically significant (p > 0.05). The difference between SCL-90-R depression subscale subscores of patients having chronic LBP and healthy persons was not statistically significant (p > 0.05). The difference between SCL-90-R somatization subscale subscores of patients having chronic LBP and healthy persons was statistically significant (p < 0.05). SCL-90-R somatization subscale subscores are higher in patients with low back pain. The treatment of low back pain can be more successful when combined with the treatment of somatization.


Skin Erythema, Pigmentation and Hydration Kinetics after Ultraviolet Radiation-induced Photodamage in Southern Chinese Women.
Although there have been some studies about changes of skin erythema and pigmentation following ultraviolet radiation in other races, the relevant data in Chinese have never been achieved. Thus, we evaluated the long-time course of skin erythema, pigmentation and hydration changes after different doses of solar-simulated ultraviolet (SSUV) irradiation in 26 Chinese women for 168 days. The erythema index increased abruptly and peaked during 3 days of SSUV exposure, then slowly returned to the baseline level starting at day 7 and completely recovered during 168-day course of this study only in one minimal erythema doses (MED) SSUV irradiation. The melanin index started to slowly increase at day 3 of SSUV exposure, peaking at day 14 and gradually returned to the baseline level thereafter, but did not return to the baseline level during 168-day course in all doses. Skin hydration slowly declined at day 3 of exposure, hitting the lowest point at day 7, then slowly recovered starting at day 14 and completely returned to the baseline level at day 28 only in 1.5MED. These results will serve as baseline data on Chinese skin and provide useful references for the treatment of serious skin photodamage in Chinese.


Delayed treatment of a posterior cruciate ligament tibial insertion avulsion fracture in a child with open physis: a case report with a 4-year follow-up.
Posterior cruciate ligament avulsion fracture from its tibial insertion is a rare pathology in children. It is usually caused by high-energy traumas and its diagnosis is not always easy. Late diagnosis leads to late treatment, which can result in suboptimal results. We present a case of a 13-year-old boy hit by a car, who had a delay in diagnosis and treatment because of severe head trauma. The patient was treated 4 months after the trauma with open reduction and internal fixation of the avulsion fracture. After 4 years of follow-up, he has no complaints and has achieved good functional outcome.


Alterations in the MA and NC domains modulate phosphoinositide-dependent plasma membrane localization of the Rous sarcoma virus Gag protein.
Retroviral Gag proteins direct virus particle assembly from the plasma membrane (PM). Phosphatidylinositol-(4,5)-bisphosphate [PI(4,5)P(2)] plays a role in PM targeting of several retroviral Gag proteins. Here we report that depletion of intracellular PI(4,5)P(2) and phosphatidylinositol-(3,4,5)-triphosphate [PI(3,4,5)P(3)] levels impaired Rous sarcoma virus (RSV) Gag PM localization. Gag mutants deficient in nuclear trafficking were less sensitive to reduction of intracellular PI(4,5)P(2) and PI(3,4,5)P(3), suggesting a possible connection between Gag nuclear trafficking and phosphoinositide-dependent PM targeting.


AUTOMATING ASSET KNOWLEDGE WITH MTCONNECT.
In order to maximize assets, manufacturers should use real-time knowledge garnered from ongoing and continuous collection and evaluation of factory-floor machine status data. In discrete parts manufacturing, factory machine monitoring has been difficult, due primarily to closed, proprietary automation equipment that make integration difficult. Recently, there has been a push in applying the data acquisition concepts of MTConnect to the real-time acquisition of machine status data. MTConnect is an open, free specification aimed at overcoming the "Islands of Automation" dilemma on the shop floor. With automated asset analysis, manufacturers can improve production to become lean, efficient, and effective. The focus of this paper will be on the deployment of MTConnect to collect real-time machine status to automate asset management. In addition, we will leverage the ISO 22400 standard, which defines an asset and quantifies asset performance metrics. In conjunction with these goals, the deployment of MTConnect in a large aerospace manufacturing facility will be studied with emphasis on asset management and understanding the impact of machine Overall Equipment Effectiveness (OEE) on manufacturing.


Ranibizumab for treatment of choroidal neovascularization secondary to age-related macular degeneration.
To evaluate the short-term outcomes after intravitreal ranibizumab (Lucentis; Genentech, Inc., South San Francisco, CA) injection in patients with neovascular age-related macular degeneration. A review of data for consecutive patients who received intravitreal ranibizumab injection was conducted. The main outcome measures were mean visual acuity and central macular thickness at 3 months compared with those at baseline. Response to ranibizumab therapy was evaluated with particular attention to prior treatment with bevacizumab (Avastin; Genentech, Inc.). Mean baseline visual acuity of 231 eyes of 231 patients was 20/152, and 189 patients (81.8%) had undergone prior treatment, with 153 (65.4%) having received intravitreal bevacizumab. Mean visual acuity at 3 months, available for 203 patients (88%), was 20/126 (P = 0.004). Mean visual acuity for 98 patients treated with bevacizumab within 3 months before ranibizumab injection was 20/100 at baseline and 20/98 at 3 months (P = 0.35). Mean baseline central macular thickness was 278 microm for all patients and improved to 211 microm at 3 months (P < 0.001). Macular thickness decrease was noted irrespective of previous bevacizumab therapy. Ranibizumab therapy was associated with significant improvements in mean visual acuity and central macular thickness for the group of all patients. Patients who had received bevacizumab treatment within 3 months before initiating ranibizumab treatment had stability of, but no improvement in, visual acuity.


Peripheral ARtery Atherosclerotic DIsease and SlEep disordered breathing (PARADISE) trial - protocol for an observational cohort study.
Peripheral arterial disease (PAD) is in fact a group of disease entities with different symptoms and course but a common underlying cause, i.e. atherosclerosis. Atherosclerosis is known to be aggravated by several cardiovascular risk factors, including obstructive sleep apnoea (OSA). Following paper is a protocol for the Peripheral ARtery Atherosclerotic DIsease and SlEep disordered breathing (PARADISE) trial, which aims to describe the prevalence of OSA in PAD patients scheduled for revascularisation, and to determine the effect of OSA on the procedure outcomes. The PARADISE study is an observational cohort trial. It plans to include 200 consecutive patients hospitalised for revascularisation due to PAD. In every patient an overnight sleep study will be performed to diagnose sleep disorders. Accord¬ing to the results of the test, patients will be divided into two groups: group A - patients with OSA, and group B - patients without OSA (control group). All patients will also be screened for classical and non-classical cardiovascular risk factors. In some of the patients, during surgery, a fragment of atherosclerotic plaque will be collected for further testing. Patients will be followed for one year for adverse events and end-points. Primary end-point of the study will be the failure of revascularisa¬tion defined as recurrence or new onset of the symptoms of ischaemia from the treated region, a need for re-operation or procedure revision, or recurrence of ischaemia signs on the imaging tests. The data obtained will help determine the incidence of OSA in the population of patients with PAD. The au¬thors expect to show that, as with other cardiovascular diseases associated with atherosclerosis, also in patients with PAD the incidence of undiagnosed OSA is high and its presence is associated with elevated cholesterol, inflammatory markers, and higher prevalence of arterial hypertension and poor control of other cardiovascular risk factors. In addition, due to increased oxidative stress and vascular endothelial injury associated with OSA, patients afflicted with this condition will not only have more advanced atherosclerotic lesions, but also in their histopathological examination their atherosclerotic plaque will exhibit evidence of greater instability and adverse morphology. We also expect to show that in patients with OSA, achieving cor¬rect control of cardiovascular risk factors will be more difficult. The study may improve PAD control through assuring better multispecialty care in PAD patients.


Mechanism of double ventricular response to a single atrial extrastimulus in patients with Wolff-Parkinson-White syndrome.
The purpose of this study is to elucidate electrophysiological determinants of double ventricular response (DVR) to a single atrial extrastimulus in Wolff-Parkinson-White (WPW) syndrome. DVR was observed in 5 (3.4%) out of 146 consecutive patients with WPW syndrome. The site of accessory pathway was located in left lateral free wall in four patients and posterior septum in one. DVR was induced by extrastimulus from coronary sinus in four patients with left-sided accessory pathway, and from both coronary sinus and high right atrium in a patient with septal accessory pathway. However, it was not possible to induce DVR from high right atrium in patients with left-sided accessory pathway, because 50 to 80 ms are needed for intra-atrial conduction from high right atrium to coronary sinus. Critical prolongation of normal AV conduction allowing DVR was seen in the slow pathway of AV node in four patients. In the remaining patients requisite conduction delay occurred in both AV node and His-Purkinje system. Single right ventricular extrastimulus could easily elicit orthodromic AV reciprocating tachycardia or echo beat in four out of five patients and incremental ventricular stimulation induced it in the remaining patient, indicating the presence of retrograde block in the normal AV pathway. As requisites of DVR to a single atrial extrastimulus in WPW syndrome: (1) slow antegrade conduction and retrograde block in the normal AV pathway; and (2) stimulation site in the vicinity of accessory pathway, are needed.


Expression of the cyclin kinase inhibitor, p27kip1, in developing and mature human kidney.
It has been shown that glomerular visceral epithelial cells (VEC) proliferate during glomerulogenesis, but differentiated VEC of the fetal kidney do not. It is also recognized that the proliferative capacity of the VEC in mature kidneys is very limited, and according to some investigators, may be completely absent. The basis for this remains unknown. Cell proliferation is controlled by cell cycle-related proteins, of which one class, the cyclin kinase inhibitors (CKI), cause cell cycle arrest and inhibit proliferation. A role for CKI in kidney development is not known. Accordingly, we examined the expression of the CKI p27kip1 (p27) in developing and mature human kidney tissue. Concomitant expression of markers of cell proliferation, Ki-67-related antigen (Ki-67) and proliferating cell nuclear antigen (PCNA), also were examined in fetal and mature human kidney tissue by immunocytochemical techniques. In developing kidney, Ki-67 and PCNA expression are most pronounced in the nephrogenic zone where expression correlates inversely with increasing glomerular maturation. In well-differentiated glomeruli, Ki-67 and PCNA expression is present in some parietal epithelial cells but is absent in the VEC. In contrast, p27 staining exhibits a reverse gradient of expression. p27 is absent in the proliferating tissue exhibiting the earliest stages of differentiation, whereas expression is widespread in the differentiated epithelial cells of more mature glomeruli, in which detectable cell proliferation has ceased. Expression of p27 was not identified in fetal mesangial or glomerular endothelial cells. In the mature human kidney, the pattern of p27 expression identified in differentiated fetal glomeruli persists and appears to be constitutive and specific for glomerular VEC. This pattern of p27 expression in terminally differentiated VEC may explain their limited proliferative capacity in response to injury. This is the first demonstration of a potential role for p27 in human renal development.


Tumor necrosis factor-alpha strongly potentiates interleukin-3 and granulocyte-macrophage colony-stimulating factor-induced proliferation of human CD34+ hematopoietic progenitor cells.
Previous studies have shown that tumor necrosis factors (TNFs) inhibit the proliferative effects of crude or purified colony-stimulating factors (CSFs) on low density human bone marrow cell fractions. In the present study we investigated the effects of TNF alpha on the growth of highly purified CD34+ human hematopoietic progenitor cells (HPC) in response to recombinant CSFs. In short-term liquid cultures (5 to 8 days), TNF alpha strongly potentiates interleukin-3 (IL-3) and granulocyte-macrophage-CSF (GM-CSF)-induced growth of CD34+ HPC, while it has no proliferative effect per se. Within 8 days, the number of viable cells obtained in TNF alpha-supplemented cultures is threefold higher than in cultures carried out with IL-3 or GM-CSF alone. Secondary liquid cultures showed that the potentiating effect of TNF alpha on IL-3-induced proliferation of CD34+ HPC does not result from an IL-3-dependent generation of TNF alpha responsive cells. Limiting dilution analysis indicates that TNF alpha increases both the frequency of IL-3 responding cells and the average size of the IL-3-dependent clones. The potentiating effect of TNF alpha on IL-3- and GM-CSF-dependent growth of CD34+ HPC is also observed in day 7 colony assays. Under these short-term culture conditions, TNF alpha does not appear to accelerate cell maturation as a precursor morphology is retained. Finally, TNF alpha inhibits the relatively weak growth-promoting effect of granulocyte-CSF (G-CSF), which acts on a more committed subpopulation of CD34+ HPC different from that recruited by IL-3 and GM-CSF. TNF beta displays the same modulatory effects as TNF alpha. Thus, TNFs appear to enhance the early stages of myelopoiesis.


Automated mechanical peripheral stimulation and postural control in subjects with Parkinson's disease and freezing of gait: a randomized controlled trial.
Individuals with Parkinson's disease (PD) and freezing of gait (FOG) have impaired postural control. Recent studies using foot sensory stimulation delivered by means of automated mechanical peripheral stimulation (AMPS) have demonstrated improvements of gait in individuals with PD. This study aimed to investigate the effects of AMPS on postural control in individuals with PD and FOG. Thirty-three subjects participated in this randomized controlled trial. Participants were allocated to two groups: AMPS and AMPS SHAM. Subjects underwent eight sessions of real (AMPS) or placebo AMPS (AMPS SHAM) once every three/four days. Postural control was assessed by means of posturography before the first and after the eighth session of treatment. We did not find positive effects of AMPS on center of pressure parameters. Thus, it seems that AMPS has no positive effect in terms of improving static postural control in individuals with PD and FOG.


[A gastroenterology unit experience with adjuvant chemotherapy for non-metastatic colon cancer: results from 24 cases].
A prospective study was carried from January 1996 to December 2000 including patients that received adjuvant chemotherapy type FUFOL for colon carcinoma after curative surgery. Chemotherapy was recommended for all stage Dukes B2 end C. Adjuvant chemotherapy was administered to 24 patients (13 men and 11 women, mean age 56.7 +/- 11.5 years) with surgically resected stage B 2 in 7 cases (29%), C in 17 cases (71%). Treatment was completed in 15 patients (62%). With a median follow-up of 31.6 months, 11 patients had no recurrence, one patient had locoregional recurrence and 2 patients developed liver metastasis. Adjuvant FUFOL chemotherapy is actually the recommended adjuvant post-surgical treatment for colon cancer. This valided protocol is easily realised with a low toxicity and it can be done in gastro-enterology unit.


Serum MicroRNAs as Diagnostic Biomarkers for Macrosomia.
Macrosomia is defined as an infant's birth weight of more than 4000 g. Although microRNAs (miRNAs) have been implicated in the pathogenesis of various diseases, the associations between serum miRNAs and macrosomia have been rarely reported. We used the Taqman Low Density Array followed by quantitative reverse transcriptase polymerase chain reaction assays to screen for miRNAs associated with macrosomia using serum samples collected 1 week before delivery. Profiling results showed that 1 miRNA was significantly upregulated and 10 miRNAs were significantly downregulated in serum samples of macrosomia (ΔΔCt > 3-fold). The expression levels of miR-21 were significantly decreased in macrosomia as compared to the controls in the third trimester. Receiver operating characteristic (ROC) curve analyses showed that the area under the ROC curve for miR-21 was 67.7% (sensitivity = 66.7% and specificity = 70.0%). miR-21 in maternal serum is differentially expressed between macrosomia and controls, and miR-21 could be used as a candidate biomarker to predict macrosomia.


[Identification of a new Puumala hantavirus subtype in rodents from China].
In order to detect the nucleic acid of Puumala hantavirus, RNA was extracted from lungs of bank voles captured in Northeast China, and partial S and M genome segments of Puumala virus were amplified by RT-PCR and sequenced. Phylogenetic analysis suggested that Chinese Puumala virus had diverged from the common node of PUUV, with accumulating nucleotide substitutions and formed a distinct lineage from other Puumala viruses. Newly found Puumala virus was most closely related to the Kamiiso-8Cr-95 and Tobetsu-60Cr-93 strains which came from Japan and the muju strains which came from South Korea. By analysis of S and M genome segments of Puumala virus, we deduced a new Puumala virus subtype did exist in Northeast China.


Structural and catalytic roles of the human 18S rRNA methyltransferases DIMT1 in ribosome assembly and translation.
rRNA-modifying enzymes participate in ribosome assembly. However, whether the catalytic activities of these enzymes are important for the ribosome assembly and other cellular processes is not fully understood. Here, we report the crystal structure of WT human dimethyladenosine transferase 1 (DIMT1), an 18S rRNA N6,6-dimethyladenosine (m26,6A) methyltransferase, and results obtained with a catalytically inactive DIMT1 variant. We found that DIMT1+/- heterozygous HEK 293T cells have a significantly decreased 40S fraction and reduced protein synthesis but no major changes in m26,6A levels in 18S rRNA. Expression of a catalytically inactive variant, DIMT1-E85A, in WT and DIMT1+/- cells significantly decreased m26,6A levels in 18S rRNA, indicating a dominant-negative effect of this variant on m26,6A levels. However, expression of the DIMT1-E85A variant restored the defects in 40S levels. Of note, unlike WT DIMT1, DIMT1-E85A could not revert the defects in protein translation. We found that the differences between this variant and the WT enzyme extended to translation fidelity and gene expression patterns in DNA damage response pathways. These results suggest that the catalytic activity of DIMT1 is involved in protein translation and that the overall protein scaffold of DIMT1, regardless of the catalytic activity on m26,6A in 18S rRNA, is essential for 40S assembly.


Cross-sectional study of the susceptibility of Candida isolates to antifungal drugs and in vitro-in vivo correlation in HIV-infected patients.
To investigate (1) the frequency of clinical resistance to oral polyenes or azole treatment for oral candidiasis, (2) the frequency of resistant in vitro Candida strains, (3) the relationship between the susceptibilities of in vitro Candida species and in vivo status in HIV patients. Prospective cross-sectional study. Tertiary care clinic at Bocage Hospital, Dijon, France. HIV-infected patients with and without oral candidiasis. Clinical examination, oral swab for mycologic investigations. Clinical diagnosis of oral candidiasis, identification of the antifungal treatment given within the previous month, identification of Candida species, antimycogramm and determination of the minimal inhibitory concentration (MIC) for fluconazole, and measurement of T-helper cell count. Within a 2-month period, 154 HIV-infected patients were studied: 46 heterosexuals, 51 intravenous drug users (IVDU), 52 homosexuals and five blood transfusion recipients. The percentages of patients with oral candidiasis were: 41, 80, 44 and 20%, respectively (P < 0.05); the mean T-helper cell counts were 200, 135, 210 and 238 x 10(6)/l cells, respectively (P < 0.05). Twenty-two patients (14.3%) had received recent azole treatment and 29 (18.8%) recent oral polyene treatment. Among the 84 patients with and the 70 patients without oral candidiasis, 78 and 28 Candida strains were isolated, respectively. Although Candida albicans represented the majority of Candida species (88 strains, 83%), the non-albicans strains were isolated more frequently in patients who had received recent antifungal treatment. No strains were resistant to ketoconazole, miconazole or econazole; however, six (5.6%), 16 (15%) and 10 (9.5%) were intermediately susceptible to the three drugs, respectively. Twelve (13.6%) of the 88 C. albicans, five of the six C. (Torulopsis) glabrata, one of the five C. tropicalis and all three C. krusei strains were resistant to fluconazole. These resistant strains were separated as follows: 41.1% of C. albicans strains resistant to fluconazole were isolated from patients who had received recent azole therapy, 17.6% from patients who received recent oral polyene, and 3.7% from patients who had not received any recent antifungal treatment (P = 0.004). The mean MIC of these three categories of isolates were 3.6, 1.6 and 0.6 mg/l, respectively (P = 0.06). Oral candidiasis and fluconazole-resistant Candida isolates are more frequently found in IVDU. Treatments using azoles select non-albicans strains and are associated with decreased susceptibilities of C. albicans strains to fluconazole in particular. These findings show that prolonged azole treatment in severely immunocompromised patients should be avoided.


Pharmacists' perceptions regarding the impact of hydrocodone rescheduling on prescription volume, workflow management, and patient outcomes.
To determine (1) pharmacists' perceptions of how rescheduling of hydrocodone combination products (HCPs) from Drug Enforcement Agency (DEA) Schedule III to DEA Schedule II has influenced prescription volume and revenue, pharmacy workflow management, and patient outcomes; and (2) whether perceptions differed between pharmacists who support versus those who oppose HCP rescheduling. A cross-sectional mail survey. Texas community pharmacies from October to December 2015. One thousand randomly selected, registered Texas community pharmacists drawn from the Texas State Board of Pharmacy registry. Pharmacists' perceptions, measured on a 5-point Likert scale of HCP rescheduling and its impact on prescription volume and revenue, workflow management, and patient outcomes. Measures were developed specifically for this study. The response rate was 17% (n = 164). The majority of pharmacists (70.4%) supported HCP rescheduling. More than 80% of respondents perceived that the volume of 2 alternative pain medications-tramadol (DEA Schedule IV) and acetaminophen with codeine (DEA Schedule III) prescriptions dispensed-either "increased" or "significantly increased" (82.0% and 85.8%, respectively) following rescheduling. Overall, pharmacists who opposed rescheduling were significantly more likely to report negative perceptions regarding revenue (P = 0.0142), inventory management (P = 0.0024), and drug shortages (P = 0.0005) than those who supported rescheduling. However, pharmacists who supported rescheduling had more positive perceptions about electronic prescribing (P <0.0115), patient safety (P <0.001), drug abuse (P <0.0001), and legitimate use (P <0.0001). Results showed that legislative efforts, such as rescheduling HCPs, influenced pharmacists' perceptions of practice and patient outcomes. Currently, little is known regarding the impact of HCP rescheduling on pharmacy practice. As new laws are passed to address the opioid epidemic in America, more research will be needed to determine whether legislation is an effective means for managing appropriate access to HCPs and other narcotic analgesics.


Cutaneous infection with Alternaria tenuissima in an immunocompromised patient.
A 28-year-old man, on immunosuppressive therapy for a recent kidney-pancreas transplant, presented to our department with an unusual violaceous nodule on his right knee. A biopsy specimen showed a granulomatous response in the dermis, surrounding numerous fungal cells and septate hyphae. Fungal cultures yielded dark grey colonies, which were identified as Alternaria tenuissima. To our knowledge this is the first report of A. tenuissima infection reported in the U.K.


Biology and mechanics of facial asymmetries in children and youths.
There is a need for further knowledge about the normal and abnormal growth patterns of bones and teeth. Genetic make-up, as well as various types of diseases and injuries such as trauma, inflammation, radiation and chemicals may affect skeletal and dental growth sites, thereby causing faulty growth. The degree of the subsequent deformity depends not only on the type, intensity, extent and chronology of the noxious agent, but also on the site and its particular involvement in growth activity. The knowledge of what happens to the stomatognathic system when some of its parts are altered is very important. Dentists are challenged to visualize the entire system in all its relationships: they must visualize the dynamics of how the system works. The relationship of the maxilla to the mandible and the mechanics of the stomatognathic system must be considered as a whole when considering the occlusion. The aim of this paper is to focus attention on the biologic basis of facial asymmetries and the functional reasons for their stabilization.


The effects of smartphone multitasking on gait and dynamic balance.
[Purpose] This study was performed to analyze the influence of smartphone multitasking on gait and dynamic balance. [Subjects and Methods] The subjects were 19 male and 20 female university students. There were 4 types of gait tasks: General Gait (walking without a task), Task Gait 1 (walking while writing a message), Task Gait 2 (walking while writing a message and listening to music), Task Gait 3 (walking while writing a message and having a conversation). To exclude the learning effect, the order of tasks was randomized. The Zebris FDM-T treadmill system (Zebris Medical GmbH, Germany) was used to measure left and right step length and width, and a 10 m walking test (10MWT) was conducted for gait velocity. In addition, a Timed Up and Go test (TUG) was used to measure dynamic balance. All the tasks were performed 3 times, and the mean of the measured values was analyzed. [Results] There were no statistically significant differences in step length and width. There were statistically significant differences in the 10MWT and TUG tests. [Conclusion] Using a smartphone while walking decreases a person's dynamic balance and walking ability. It is considered that accident rates are higher when using a smartphone.


Ratio of ellipticities between 192 and 208 nm (R1 ): An effective electronic circular dichroism parameter for characterization of the helical components of proteins and peptides.
Circular dichroism (CD) spectroscopy represents an important tool for characterization of the peptide and protein secondary structures that mainly arise from the conformational disposition of the peptide backbone in solution. In 1991 Manning and Woody proposed that, in addition to the signal intensity, the ratio between [θ]nπ* and [θ]ππ*ǁ ((R2 ) ≅ [θ]222 /[θ]208 ), along with [θ]ππ*⊥ and [θ]ππ*ǁ ((R1 ) ≅ [θ]192 /[θ]208 ), may be utilized towards identifying the peptide/protein conformation (especially 310 - and α-helices). However, till date the use of the ratiometric ellipticity component for helical structure analysis of peptides and proteins has not been reported. We studied a series of temperature dependent CD spectra of a thermally stable, model helical peptide and its related analogs in water as a function of added 2,2,2-trifluoroethanol (TFE) in order to explore their landscape of helicity. For the first time, we have experimentally shown here that the R1 parameter can characterize better the individual helices, while the other parameter R2 and the signal intensity do not always converge. We emphasize the use of the R1 ratio of ellipticities for helical characterization because of the common origin of these two bands (exciton splitting of the amide π→ π* transition in a helical polypeptide). This approach may become worthwhile and timely with the increasing accessibility of CD synchrotron sources.


Complete pathological response of multiple huge liver metastases of colon cancer: a case report.
We report a case of a pathological complete response (pCR) with chemotherapy for multiple huge liver metastases from colon cancer. A 59-year-old woman presented with anorexia and weight loss. Laboratory tests revealed elevated liver enzyme levels and tumor markers. A computed tomography/positron emission tomography-computed tomography scan revealed a transverse colon tumor and unresectable liver masses measuring 9.0 cm in maximum diameter in segments 7 and 8, with another mass in segment 6. She underwent laparoscopic colectomy and was administered FOLFOX + BV. After 11-cycles of chemotherapy, the liver masses became resectable with a partial response, so hepatectomy was performed. On the final histopathological analysis, all lesions were fibrotic without any viable cancer cells. The patient is alive without recurrence 2 years after resection. We believe this is the largest tumor of unresectable colorectal liver metastasis (CRLM) that has ever resulted in pCR with chemotherapy. FOLFOX + Bev was thus found to be an effective treatment for unresectable CRLM.


Stress echocardiography.
Stress echocardiography has a number of advantages over exercise electrocardiography. It has superior sensitivity and specificity as it detects wall motion abnormalities, an earlier and more specific marker for myocardial ischaemia than ST segment depression. It can also localise ischaemia and identify hibernating myocardium. Compared with radionuclide imaging, stress echocardiography has similar accuracy but is cheaper and does not involve exposure to ionising radiation. The disadvantage of stress echocardiograph is that it requires expertise and specialist experience of at least 100 studies. Current research aims at improving objective methods of analysing results. The technique is justifiably being adopted throughout the UK.


The Relationship between Serum and Gene Expression Levels of RANK, RANKL and Osteoprotegerin Inflammatory Pathway with Unstable Angina: A Case-control Study.
Osteoprotegerin (OPG), receptor activator of nuclear factor-kappa B (RANK) and receptor activator of nuclear factor-kappa B ligand (RANKL), the members of the tumor necrosis factor (TNF) family, have multiple effects on bone metabolism, endocrine functions and, as an inflammatory pathway, in the immune system. This study tried to determine the association of the OPG/RANKL/RANK axis with the severity of unstable angina (UA) as an inflammatory condition. Our study involved 50 patients with UA and 50 healthy people. Serum and peripheral blood mononuclear cells were isolated from all participants. Serum levels and gene expression of OPG, RANKL, and RANK in mononuclear cells were measured by enzyme-linked immunosorbent assay (ELISA) and real-time polymerase chain reaction (RT-PCR), respectively. For each patient with UA, the thrombolysis in myocardial infarction (TIMI) and the global registry of acute coronary events (GRACE) scores were determined to evaluate the severity of the disease. Then we analyzed the relation of OPG, RANKL, and RANK levels with TIMI and GRACE scores in patients with UA. Discriminate analysis was used to predict the combinational models of such factors on the prediction of UA. Serum levels of OPG and RANKL (p<0.001) and gene expression of RANKL (p<0.001) were significantly more in patients than those in healthy ones. No relation was seen between the OPG/RANKL/RANK axis and the severity of UA according to TIMI and GRACE scores. Our study shows that serum level, as well as gene expression of OPG/RANKL/RANK axis neither, predicts the occurrence of UA nor shows any relationship with its severity.


The relevance of cardio-pulmonary-vascular reflex to regulation of the brain vessels.
In 11 cordotomized (C2) rats with the vagi cut, monopolar electrical stimulation at a current intensity of 73 +/- 21 (S.E.) microA (n = 6) to the intermediate region of the solitary nucleus elicited an increase (p less than 0.05) in regional blood flow (iodoantipyrine technique) of 71% and 43% in the frontal cortex and caudate nucleus, respectively. The findings suggest that some of the cardio-pulmonary and cardio-vascular reflex mechanisms are involved in the regulation of the blood vessels of the brain.


Assessment of Sitting in Children with Cerebral Palsy from Videofilm.
The purpose of this study was to investigate the reliability of two dependent measures, (1) counting of pathologic movements (CPM) and (2) the Sitting Assessments Scale (SAS). The measures were intended for use in clinical physiotherapy practice, to provide both quantifiable and qualitative information from videofilm concerning functional sitting in children with cerebral palsy (CP). Two separate groups of final term physiotherapy students (three students in each group) tested one technique each for interrater reliability. The SAS-measure also was tested for interrater reliability. Videofilm of twelve children with CP ranging in age from 2 to 16 years were assessed. The students' results were compared to earlier results by experiences physiotherapist. The results demonstrated excellent interrater reliability for SAS (p<0.001) and good interrater reliability for the CPM technique. The degree of user experience seems to play a role in the use of the CPM measure. Considering evaluation of postural control and functional performance over all, SAS appears to be superior to CPM.


Effectiveness of physical activity as primary preventive care for lower urinary tract symptoms in elderly people through the "Muscle Enhancing Club".
[Purpose] The aim of this study was to examine the relationship between physical activity (PA) in elderly people as preventive exercise for lower urinary tract symptoms (LUTS) or overactive bladder (OAB), and how PA in primary preventive care can impact change upon LUTS. [Subjects and Methods] An interview sheet featuring LUTS domains (IPSS/QOL and OABSS questionnaire) was distributed to all participants (104 males and 494 females) who attended the public elderly people's physical exercise class "Muscle Enhancing Club 2010" in Asahikawa city. [Results] The interview sheet was collected from 65 males (75.4 ± 5.79 years of age) and 304 females (72.7 ± 5.23 years of age). In all cases, there was a statistically significant difference of QOL. In 77 LUTS cases, there was a statistically significant difference in the change of the following LUTS domains: incomplete bladder emptying, frequency, nocturia, QOL, urgency, and OABSS total score. In 61 OAB cases, there was a statistically significant difference in the change of the following LUTS domains: QOL, urgency, and OABSS total score. [Conclusion] PA, as a preventative exercise for elderly people, led to improvements in LUTS, especially storage symptoms and QOL. Our data show that gross movement and education was beneficial. From the point of preventive medicine, various exercise classes, organized by the local government and others, also indicated that these classes increase the possibility of improvement to LUTS.


A rapid, noninvasive method for evaluation of total menstrual loss.
Three types of packages containing enough sanitary supplies for one menstrual period were weighed and distributed to 100 subjects to determine adequacy of supplies and compliance to protocol. Returned packages were then reweighed to determine total menstrual loss. No subject used all the supplies in her package; therefore, packages contents were deemed ample. 7 subjects added their own purchased products to their packages; since added products were specified by name, weight corrections were easily made. 2 subjects lost unused supplies which were specified, and weight corrections were made accordingly. Evaporative loss from sealed and stored and frequently opened packages was measured. 100 g distilled water were added to contents inside 60 packages which were sealed and set aside 7 days. Average loss from these packages was 0.65 +/- 0.57 g. 5 g water were added to 60 packages 4 times daily for 5 days (100 g total). Then packages were set aside for 2 days and weighed on day 7. Average loss from these packages was 1.25 +/- 0.85 g. The combination of storage plus frequent opening resulted in an average loss of 1.90 +/- 0.87 g. Thus, the direct-weight method permits recovery of 97-98% of sample.


Demographic Characteristics and Disease Pattern Among Non-COVID-19 Patients Admitted to Medical Units of a Tertiary Care Hospital in Sri Lanka During Lockdown of the COVID-19 Pandemic: A Comparison During and Before Lockdown.
The COVID-19 infection was rapidly spreading almost all over the world, and the first case was confirmed on 27th January 2020 by a foreign tourist in Sri Lanka. The first Sri Lankan citizen with COVID-19 was confirmed on 11th March 2020. Soon after the confirmation of the disease, long days of lockdown were imposed in almost all parts of the world, including Sri Lanka, to control the spread of the disease. Objectives: To determine the demographic characteristics such as age, sex, number of patients, and disease pattern among non-COVID-19 patients admitted to the medical units during the lockdown of the COVID-19 pandemic and to compare these characteristics with the data before the lockdown. This was a cross-sectional analytical study. It was conducted at the Teaching Hospital in Batticaloa, Sri Lanka. All the non-COVID-19 patients admitted to medical wards and intensive care units (ICU) were included in this study. Patients admitted to the medical intensive care unit (MICU) and coronary care unit (CCU) were considered ICU admissions in this study. They were studied over a period of one month during lockdown (11th March 2020 to 10th April 2020) and compared with the patients admitted one month prior to the lockdown (11th February 2020 to 10th March 2020). Totally, 2340 non-COVID-19 patients (52.5% males) were admitted before the lockdown, and 1376 non-COVID-19 patients (56.2% males) were admitted during the lockdown. This reduction in admission is statistically significant (p-value is <0.001, df=3715). Patients admitted to the wards before lockdown were 2283 (97.6%) and during lockdown were 1309 (95.1%). ICU admissions were N=57 (2.4%) before lockdown and N=67 (4.9%) during lockdown. The common age distribution before the lockdown showed that 26.4% were 31-50 and 41.5% were 51-70 years. Similarly, during lockdown, the age distribution disclosed that 28.9% were 31-50 years and 42.9% were 51-70 years. The disease pattern demonstrated that before lockdown, the majority of patients were admitted for routine hemodialysis (13.2%), to get an injection (9.9%), ischemic heart disease (8.4%), chronic kidney disease (7.3%), and viral fever, including dengue (7.2%). Likewise, during lockdown, more patients were admitted for routine hemodialysis (10.7%), viral fever, including dengue (9.3%), ischemic heart disease (8.8%), to get an injection (8.5%), and chronic kidney disease (5.9%). Conclusion: There was a significant reduction in the number of non-COVID-19-related admissions during the period of lockdown. However, there was not much difference in the proportion of admissions according to gender, age, and disease pattern before and during lockdown. More number of male patients were admitted than female patients. Most of the admitted patients were under the age group of 51-70 years. The highest number of patients were admitted for routine hemodialysis before and during lockdown. However, a slightly higher number of patients were admitted to the ICU during lockdown. Therefore, strengthening the ICU facilities may be an important preparation to accommodate more patients in the future if a similar kind of emergency lockdown occurs in a pandemic situation. In addition, admissions due to non-communicable diseases (NCD) didn't fall in proportion during the pre-COVID-19 period and the lockdown period. Therefore, the redistribution of healthcare facilities needs to be done wisely to face the challenges caused by the NCDs.


FA2H-dependent fatty acid 2-hydroxylation in postnatal mouse brain.
2-Hydroxy fatty acids are relatively minor species of membrane lipids found almost exclusively as N-acyl chains of sphingolipids. In mammals, 2-hydroxy sphingolipids are uniquely abundant in myelin galactosylceramide and sulfatide. Despite the well-documented abundance of 2-hydroxy galactolipids in the nervous system, the enzymatic process of the 2-hydroxylation is not fully understood. To fill this gap, we have identified a human fatty acid 2-hydroxylase gene (FA2H) that is highly expressed in brain. In this report, we test the hypothesis that FA2H is the major fatty acid 2-hydroxylase in mouse brain and that free 2-hydroxy fatty acids are formed as precursors of myelin 2-hydroxy galactolipids. The fatty acid compositions of galactolipids in neonatal mouse brain gradually changed during the course of myelination. The relative ratio of 2-hydroxy versus nonhydroxy galactolipids was very low at 2 days of age ( approximately 8% of total galactolipids) and increased 6- to 8-fold by 30 days of age. During this period, free 2-hydroxy fatty acid levels in mouse brain increased 5- to 9-fold, and their composition was reflected in the fatty acids in galactolipids, consistent with a precursor-product relationship. The changes in free 2-hydroxy fatty acid levels coincided with fatty acid 2-hydroxylase activity and with the upregulation of FA2H expression. Furthermore, mouse brain fatty acid 2-hydroxylase activity was inhibited by anti-FA2H antibodies. Together, these data provide evidence that FA2H is the major fatty acid 2-hydroxylase in brain and that 2-hydroxylation of free fatty acids is the first step in the synthesis of 2-hydroxy galactolipids.


[Example of examination of allergic substances (egg, milk) in commercial foods].
Allergic substances (eggs, milk) in commercial foods were measured by means of the notified ELISA methods using FASTKIT ELISA kit (N kit) and "Tokuteigenzairyo sokutei kit" (M kit). Some samples were also analyzed with the notified western blot method using "Tokuteigenzairyo western blot kit". In the methods for detection of eggs, proteins of fish egg and chicken muscle were examined using the ELISA and Western blotting methods to check for cross-reaction. Sujiko was false positive with the M kit. Difficulties were also encountered in the notified ELISA and Western blotting methods with raw chicken. In foods labeled as containing eggs, it was difficult to detected in the heart-processed food using both notified methods. In the method for detection of milk, foods containing thickening polysaccharides, showed cross-reaction with the N kit. However, this cross-reaction was eliminated when an improved N kit was used. Foods labeled as containing milk did not present any difficulty.


An anomaly within the Latino epidemiological paradox: the Latino adolescent male mortality peak.
To describe the anomaly of the Latino adolescent male mortality peak in relation to the overall Latino epidemiological paradox and in relation to the need for new conceptual models describing the health of a culturally diverse population. Population-based study using California's 1989 to 1997 summary death files for death-related information and the State of California Department of Finance population estimates for population denominators for corresponding years. California's general population for 1989 to 1997, including California's 15- to 19-year-old and 20- to 24-year-old populations. In 1997, those 2 age groups numbered 4.3 million. Mortality rates for Latinos and African Americans compared with non-Hispanic whites expressed as relative risk (RR). Overall, the Latino RR of mortality follows the Latino epidemiological paradox in that it is lower (RR, <1.00) than that of non-Hispanic whites for most age groups and both sexes. The anomaly within this paradox is seen in Latino males aged 15 to 19 years (RR, 1.77; 95% confidence interval, 1.55-2.02) and 20 to 24 years (RR, 1.79; 95% confidence interval, 1.58-2.02). This period of elevated mortality risk is labeled the Latino adolescent male mortality peak, and it is an anomaly within the overall Latino epidemiological paradox.


The gene and messenger RNA for adenovirus polypeptide IX.
A small RNA species, distinct from the VA RNAs, has been identified in HeLa cells infected with adenovirus type 2. The RNA, which has been purified using a novel screening procedure, is polyadenylated, sediments at 9S and has an estimated length of 550 nucleotides. In a cell-free translation system, the 9S RNA directs the synthesis of virion polypeptide IX, molecular weight 12,000 daltons. The location of its gene has been established by hybridization of the RNA to fragments of viral DNA produced by cleavage with restriction endonucleases: it spans position 10.0 on the r strand of the viral genome. These results unexpectedly place the gene for a "late" protein within a region of the genome which is transcribed early during infection


A direct thrombin inhibitor studied by dynamic whole blood clot formation. Haemostatic response to ex-vivo addition of recombinant factor VIIa or activated prothrombin complex concentrate.
Direct thrombin inhibitors have proven efficacious in prevention of venous thromboembolism. Bleeding complications are rare, but in case of acute serious bleeding, an effective and instant haemostatic intervention may be required. In the present study it was demonstrated that the direct thrombin inhibitor melagatran induces dose-dependent abnormalities in whole blood (WB) clotting profiles as recorded by a recently described modified thrombelastographic model, and that rFVIIa or APCC are capable of improving the haemostatic capacity. Experiments were performed using WB from 30 healthy males. In-vitro titration experiments (n = 10) with addition of melagatran to WB corresponding to plasma concentrations ranging from 0 to 5.0 microM (12 steps) showed a dose-dependent prolongation of the clot initiation and characteristic decrease of the maximum rate of clot propagation. In-vitro intervention studies (n = 20) were completed with four different concentrations of melagatran as well as addition of four different levels of rFVIIa or APCC. At all tested concentrations of melagatran, rFVIIa significantly shortened the melagatran-induced prolonged clot initiation but induced only minor improvements of the reduced clot propagation. In contrast, APCC significantly and dose-dependently shortened the clot initiation and accelerated the clot propagation. In conclusion, our thrombelastographic model appears useful for evaluating the effect of direct thrombin inhibitors on dynamicWB clot formation and rFVIIa, but especially APCC significantly improved theWB clot formation. The pronounced stabilizing effect of APCC may be caused by its content of prothrombin and activated coagulation factors.


Multi-energy calibration for determining critical metals in nickel-metal hydride battery residues by microwave-induced plasma atomic emission spectrometry.
Nickel-metal hydride batteries (NiMH) are a secondary source of high aggregate value elements, such as nickel, manganese, cobalt, and rare earths, for which recycling typically involves acid lixiviation. Designing the recycling process requires accurate determination of the elements in the leachates, which is hindered by the high complexity of the matrix. In the present study, microwave-induced plasma atomic emission spectrometry (MIP-OES) was selected as the quantitative method for elemental analysis because of its environment friendliness and cost-effectiveness. Multi-energy calibration (MEC) was also pioneeringly evaluated to circumvent matrix effects and simplify the determination of Ce, La, Ni, Co, and Mn in sulfuric acid leachates of NiMH batteries by MIP-OES. The method's analytical performance and accuracy were critically compared with external standard calibration and the standard additions method. MEC yielded superior results, with analyte recoveries within 90-110%, precision (coefficients of variation) from 1.8% to 5.8%, and limits of detection of 10, 20, 1, 400, and 60 μg kg-1 for Ni, La, Mn, Ce, and Co, respectively. The results demonstrated the ability of MEC-MIP-OES to minimize matrix effects, as well as simplify and speed up the analysis of NiMH battery leachates, which is compatible with this high-demand analytical application.


The protease inhibitor JO146 demonstrates a critical role for CtHtrA for Chlamydia trachomatis reversion from penicillin persistence.
The Chlamydia trachomatis serine protease HtrA (CtHtrA) has recently been demonstrated to be essential during the replicative phase of the chlamydial developmental cycle. A chemical inhibition strategy (serine protease inhibitor JO146) was used to demonstrate this essential role and it was found that the chlamydial inclusions diminish in size and are lost from the cell after CtHtrA inhibition without formation of viable elementary bodies. The inhibitor (JO146) was used in this study to investigate the role of CtHtrA for penicillin persistence and heat stress conditions for Chlamydia trachomatis. JO146 addition during penicillin persistence resulted in only minor reductions (~1 log) in the final viable infectious yield after persistent Chlamydia were reverted from persistence. However, JO146 treatment during the reversion and recovery from penicillin persistence was completely lethal for Chlamydia trachomatis. JO146 was completely lethal when added either during heat stress conditions, or during the recovery from heat stress conditions. These data together indicate that CtHtrA has essential roles during some stress environments (heat shock), recovery from stress environments (heat shock and penicillin persistence), as well as the previously characterized essential role during the replicative phase of the chlamydial developmental cycle. Thus, CtHtrA is an essential protease with both replicative phase and stress condition functions for Chlamydia trachomatis.


Intestinal heterophyidiasis: an emerging food-borne parasitic zoonosis in southern Philippines.
Heterophyidiasis is an infection of the small bowel by minute intestinal flukes of the genus Heterophyes or related members of the family Heterophyidae. To provide a better understanding of this rarely reported condition, this study attempted to determine the infection rate, intensity of infection as well as the clinical spectrum of heterophyid infection. A stool survey was conducted in barangay San Isidro, Monkayo, Compostela Valley. Thirty-six percent of patients with history of bowel disturbance (abdominal discomfort/pain and/or diarrhea) in the past 4 weeks were found to have heterophyidiasis. All age groups were infected, with the youngest patient being 1 year 7 months of age, while the oldest patient was 73 years of age. Prevalence was highest in the 15 to 30 years old group at 55.3%. The most common clinical manifestations of heterophyidiasis were signs and symptoms of acid peptic or peptic ulcer disease. Early diagnosis and treatment are important to ensure prompt resolution of heterophyid infection, hence, decreased morbidity and decreased chances for complications like heart or brain involvement. Proficiency of laboratory staff should be enhanced to ensure accurate diagnosis which will then make appropriate treatment possible.


[Origins of French telemedicine legislation].
After the creation of the european institute of telemedicine on 10 July 1989 at Paul Sabatier university in Toulouse, it became clear that legislation was needed to provide a solid framework for this new medical practice. The French government's decision in 1993 to create an experimental telemedicine network in the Midi-Pyrenees region, bringing together all public and private healthcare establishments, led us to manage telemedicine in much the same way as a hospital department. This resulted in the creation of institutional systems suited to multicenter networking and requiring particular regulations to govern healthcare professionals' responsibilities, financing, data security, and the use of new communication and information technologies. Articles specific to telemedicine were integrated into the law of 13 August 2004 relating to the reorganisation of French healthcare insurance.


MHC class II antigens on lung epithelial of human fetuses and neonates. Ontogeny and expression in lungs with histologic evidence of infection.
An immunohistochemical study with anti-HLA-DP, anti-HLA-DQ and anti-HLA-DR monoclonal antibodies was carried out on lung tissue from 29 fetuses and 19 infants that died within the 1st year of life. The aim of the study was to verify the influence of lung maturation and lung inflammation, caused by intrauterine infection, on the expression of MHC class II antigens on lung epithelia. No positive reaction of lung epithelia was noticed in 6 fetuses of less than 21 weeks' gestation. In fetuses of more than 21 weeks' gestation, lung inflammation strongly correlated with the expression of MHC class II subregion products on lung epithelia. All 4 fetuses with histopathologic evidence of lung infection but only 4 out of 23 fetuses without lung inflammation bore HLA-DR antigens on their immature cuboidal alveolar epithelium. Within the bronchial epithelial layers only scattered MHC class II positive epithelia were found. After birth, MHC class II antigens were almost constantly detected on cuboidal type II pneumocytes of inflamed and noninflamed lung tissue as well. Only 3 infants that died within the 1st week of life did not show any MHC class II antigen expression on their lung epithelia.


The determination of megakaryocyte ploidy.
Methods which have been used to determine megakaryocyte ploidy in animals and humans are reviewed. Although the number of megakaryocyte nuclear units counted in bone marrow squashes is roughly proportional to ploidy, accurate determinations of DNA content require the use of microphotometry or flow cytometry. New techniques should make it possible to distinguish polyploidizing megakaryoblasts from promegakaryocytes and mature megakaryocytes which have arrested polyploidization. Only the latter should be included in histograms of the number of endoduplications, since only those have expressed their full polyploidization potential. Statistical techniques are available for analysis and comparison of both raw ploidy distributions or histograms of endoduplication numbers.


Inhibition of the NF-κB Signaling Pathway Alleviates Pyroptosis in Bladder Epithelial Cells and Neurogenic Bladder Fibrosis.
Most children with a neurogenic bladder (NB) have bladder fibrosis, which causes irreversible bladder dysfunction and damage to the upper urinary tract. However, the mechanism of bladder fibrosis remains unclear. This study aimed to investigate the underlying causes of bladder fibrosis. Here, the lumbar 6 (L6) and sacral 1 (S1) spinal nerves of Sprague Dawley rats were severed bilaterally to establish NB models. Using RNA-seq, we discovered that the NF-κB signaling pathway and inflammation were upregulated in spinal cord injury (SCI)-induced bladder fibrosis. Subsequent Western blotting, enzyme-linked immunosorbent assays, immunohistochemical staining, and immunofluorescence staining verified the RNA-seq findings. To further clarify whether the NF-κB signaling pathway and pyroptosis were involved in bladder fibrosis, a TGF-β1-treated urinary epithelial cell line (SV-HUC-1 cells) was used as an in vitro model. Based on the results of RNA-seq, we consistently found that the NF-κB signaling pathway and pyroptosis might play important roles in TGF-β1-treated cells. Further experiments also confirmed the RNA-seq findings in vitro. Moreover, using the NLRP3 inhibitor MCC950 rescued TGF-β1-induced fibrosis, and the NF-κB signaling pathway inhibitor BAY 11-7082 effectively rescued TGF-β1-induced pyroptosis and the deposition of extracellular matrix by SV-HUC-1 cells. In summary, our research demonstrated for the first time that the NF-κB signaling pathway inhibition rescued bladder epithelial cells pyroptosis and fibrosis in neurogenic bladders.


RNA interference targeting CITRON can significantly inhibit the proliferation of hepatocellular carcinoma cells.
CITRON (rho-interacting, serine/threonine kinase 21), which is a key component of the midbody, is essential for cytokinesis. However, the role of CITRON in hepatocellular carcinoma (HCC) is poorly understood. Here we first measured the expression of CITRON in HCC specimens by quantitative real-time RT-PCR and immunohistochemical staining. The results showed CITRON to be frequently up-regulated in HCC as compared with adjacent non-tumour tissues. Then we employed adenovirus-mediated RNA interference against CITRON to assess its anti-proliferation effect on SMMC-7721 cells, a representative HCC cell line. The resulting data demonstrated that CITRON knockdown was capable of inhibiting the proliferation and colony formation of SMMC-7721 cells, with an obvious increase of multinucleated cells. Furthermore, we subcutaneously injected the SMMC-7721 cells with the CITRON knockdown into nude mice to evaluate the tumourigenicity. The data indicated that adenovirus-mediated RNA interference can suppress tumourigenicity in vivo of HCC cells. Our data suggest that CITRON may be a potential target for therapeutic intervention in HCC.


Cranial remodeling to treat craniosynostosis by gradual distraction using a new device.
The authors describe their experience in remodeling the shape of the cranium in children with craniosynostosis by using gradual distraction. In half of the cases, a new distraction device developed by the authors was used. Twenty children with craniosynostosis underwent frontoorbital advancement and cranial remodeling assisted by gradual distraction. There were five children with brachycephaly, two with oxycephaly, five with scaphocephaly, two with plagiocephaly, one with combined scaphocephaly and plagiocephaly, and five with trigonocephaly. Four cases were syndromic and 16 were nonsyndromic. The patients ranged in age from 3 to 50 months (mean 14.5 months) at the first surgery. Simulated surgery was first performed on a three-dimensional solid model made of polyurethane, which accurately represented cranial flexibility, to determine the most favorable osteotomy line. Distraction was initiated 1 week postoperatively. The speed and extent of advancement (maximum extension 45 mm) were predetermined on the basis of previously reported criteria and the results of simulated surgery. Postoperatively, the cranial configuration was favorable in all cases. Spontaneous remodeling of irregularities and/or gaps apparent after distraction was found to occur 2 to 5 months after removal of the distraction devices, especially in patients with trigonocephaly or scaphocephaly. No major perioperative complication was observed in any patient. There were minor complications in six of the first 10 cases, including exposure of the device, shaft slippage, and fluid discharge. A new device was developed and used on the last 10 patients treated; it successfully eliminated device exposure and shaft slippage. A 3-year follow-up review confirmed that there was no relapse of advanced bones. Highly satisfactory results were achieved in cases of both syndromic and nonsyndromic craniosynostosis when gradual distraction was performed.


The involvement of interleukin-22 in the expression of pancreatic beta cell regenerative Reg genes.
In Type 1 diabetes, the insulin-producing β-cells within the pancreatic islets of Langerhans are destroyed. We showed previously that immunotherapy with Bacillus Calmette-Guerin (BCG) or complete Freund's adjuvant (CFA) of non-obese diabetic (NOD) mice can prevent disease process and pancreatic β-cell loss. This was associated with increased islet Regenerating (Reg) genes expression, and elevated IL-22-producing Th17 T-cells in the pancreas. We hypothesized that IL-22 was responsible for the increased Reg gene expression in the pancreas. We therefore quantified the Reg1, Reg2, and Reg3δ (INGAP) mRNA expression in isolated pre-diabetic NOD islets treated with IL-22. We measured IL-22, and IL-22 receptor(R)-α mRNA expression in the pancreas and spleen of pre-diabetic and diabetic NOD mice. Our results showed: 1) Reg1 and Reg2 mRNA abundance to be significantly increased in IL-22-treated islets in vitro; 2) IL-22 mRNA expression in the pre-diabetic mouse pancreas increased with time following CFA treatment; 3) a reduced expression of IL-22Rα following CFA treatment; 4) a down-regulation in Reg1 and Reg2 mRNA expression in the pancreas of pre-diabetic mice injected with an IL-22 neutralizing antibody; and 5) an increased islet β-cell DNA synthesis in vitro in the presence of IL-22. We conclude that IL-22 may contribute to the regeneration of β-cells by up-regulating Regenerating Reg1 and Reg2 genes in the islets.


'It is just the way it was in the past before I went to test': a qualitative study to explore responses to HIV prevention counselling in rural Tanzania.
Voluntary counselling and testing (VCT) for HIV first evolved in Western settings, with one aim being to promote behaviours which lower the risk of onward transmission or acquisition of HIV. However, although quantitative studies have shown that the impact of VCT on sexual behaviour change has been limited in African settings, there is a lack of qualitative research exploring perceptions of HIV prevention counselling messages, particularly among clients testing HIV-negative. We conducted a qualitative study to explore healthcare worker, community and both HIV-negative and HIV-positive clients' perceptions of HIV prevention counselling messages in rural Tanzania. This study was carried out within the context of an ongoing community HIV cohort study in Kisesa, northwest Tanzania. Nine group sessions incorporating participatory learning and action (PLA) activities were conducted in order to gain general community perspectives of HIV testing and counselling (HTC) services. Thirty in-depth interviews (IDIs) with HIV-negative and HIV-positive service users explored individual perceptions of HIV prevention counselling messages, while five IDIs were carried out with nurses or counsellors offering HTC in order to explore provider perspectives. Two key themes revolving around socio-cultural and contextual factors emerged in understanding responses to HIV prevention counselling messages. The first included constraints to client-counsellor interactions, which were impeded as a result of difficulties discussing private sexual behaviours during counselling sessions, a hierarchical relationship between healthcare providers and clients, insufficient levels of training and support for counsellors, and client concerns about confidentiality. The second theme related to imbalanced gender-power dynamics, which constrained the extent to which women felt able to control their HIV-related risk. Within the broader social context of a rural African setting, HIV prevention counselling based on a Western model of individual-level agency seems unlikely to make a significant contribution to sexual behaviour change until there is greater recognition by counsellors of the ways in which power dynamics within many relationships influence behaviour change. More culturally relevant counselling strategies and messages and infrastructural improvements such as additional training for counsellors and counselling rooms which ensure privacy and confidentiality, may lead to better outcomes in terms of sexual risk reduction.


A review of translational animal models for knee osteoarthritis.
Knee osteoarthritis remains a tremendous public health concern, both in terms of health-related quality of life and financial burden of disease. Translational research is a critical step towards understanding and mitigating the long-term effects of this disease process. Animal models provide practical and clinically relevant ways to study both the natural history and response to treatment of knee osteoarthritis. Many factors including size, cost, and method of inducing osteoarthritis are important considerations for choosing an appropriate animal model. Smaller animals are useful because of their ease of use and cost, while larger animals are advantageous because of their anatomical similarity to humans. This evidence-based review will compare and contrast several different animal models for knee osteoarthritis. Our goal is to inform the clinician about current research models, in order to facilitate the transfer of knowledge from the "bench" to the "bedside."


Crystal structure and photochemical behavior in solution of the 3'-N-sulfamate analogue of thymidylyl(3'-5')thymidine.
The 3'-N-sulfamate analogue of thymidylyl(3'-5')thymidine (TnsoT, 1) exhibits a preference for a C3'-endo conformation in the solution and solid states. Its photochemical behavior in solution is compared to that of its natural counterpart, thymidylyl(3'-5')thymidine (TpT, 2), to get further insight into the significance of the C3'-endo conformation on the photoproduct formation at the single-stranded dinucleotide level. Irradiation at 254 nm of 1 led to the same type of photoproducts as observed with 2. However, 1 was significantly more photoreactive than 2, and accordingly, the initial rate of photoproduct formation was enhanced in accordance with its propensity to base stack compared to 2. The corresponding quantum yields were determined and showed that the enhancement factor (1 compared to 2) is moderate for the cyclobutane pyrimidine dimer (CPD) (1.26) and much higher for the (6-4) photoproduct (1.8). These data strongly suggest that the CPD and (6-4) photoproduct arise from distinct minor stacked conformations.


Improvement of vascular function by acute and chronic treatment with the PDE-5 inhibitor sildenafil in experimental diabetes mellitus.
Diabetes-associated vascular dysfunction contributes to increased cardiovascular risk. We investigated whether the phosphodiesterase-5 inhibitor sildenafil would improve vascular function in diabetic rats. Male Wistar rats were injected with streptozotocin (50 mg kg(-1), i.v.) to induce insulin-deficient diabetes. Direct effects of sildenafil as well as modification of endothelium-dependent and -independent vasorelaxation were investigated in vitro. The effects of acute and chronic (2 week) treatment in vivo of sildenafil on vascular function were also characterized in isolated aortic segments in organ bath chambers 4 weeks after diabetes induction. Sildenafil induced a concentration-dependent vasorelaxation, which was attenuated by the nitric oxide (NO) synthase inhibitor, N(G)-nitro-L-arginine. Acetylcholine-induced endothelium-dependent as well as endothelium-independent relaxation induced by the NO donor, DEA-NONOate, was significantly reduced in aortae from diabetic rats. Incubation with sildenafil in vitro normalized both endothelium-dependent and -independent relaxation in aortae from diabetic rats. Acute as well as chronic in vivo treatment with sildenafil resulted in enhanced endothelium-dependent and -independent vasorelaxation. Superoxide formation was increased in diabetes, associated with enhanced membrane expression of the NAD(P)H oxidase subunit gp91(phox) and Rac, which were both reduced by chronic treatment with sildenafil. We demonstrate that sildenafil treatment rapidly and chronically improves vascular relaxation in diabetic rats. Treatment with sildenafil might provide a similarly beneficial effect in diabetic patients.


An improved method for the isolation and culture of rat pancreatic ductal epithelial cells.
This aim of this study was to explore a novel method that can be used to isolate and culture rat pancreatic ductal epithelial cells. Retrograde injection of indigo carmine solution into the bile duct of rats revealed the main pancreatic duct, which was isolated using the naked eye (without using a stereomicroscope). The main pancreatic duct was sequentially digested with three enzymes, and the digested cells were cultured in RPMI-1640 medium containing 10-15% fetal bovine serum. After 72 h, the primary pancreatic ductal epithelial cells started to adhere to the wall. The cells reached 70-80% confluence after approximately 7 days and were subsequently digested with 0.25% trypsin and subcultured. Cells of the second and fourth passages were harvested. Cytokeratin (CK)-7 and CK-19 protein expressions in the cells and pancreatic tissue were detected by Western blot analysis. q-PCR was employed to examine CK-7, CK-19, somatostatin, amylase, insulin, and glucagon mRNA expression in the cells and pancreatic tissue after the main pancreatic duct was removed. The rats had one or two main pancreatic ducts meeting the bile ducts at a right or an acute angle. Rat pancreatic ductal epithelial cells isolated by this method grew well and showed a cobblestone-like appearance via microscopy. Western blot analysis showed that both the second and fourth passages of pancreatic ductal epithelial cells expressed CK-7 and CK-19 protein. The q-PCR results showed the expression of CK-7 and CK-19 genes in the second and fourth passages of pancreatic ductal epithelial cells, while the somatostatin, amylase, insulin, and glucagon genes were not expressed. The pancreatic tissue harvested after the removal of the main pancreatic duct did not express CK-7 or CK-19, while the somatostatin, amylase, insulin, and glucagon genes were expressed. The preliminary results show that this method can be applied to successfully isolate and culture rat pancreatic ductal epithelial cells.


Rotational molecular dynamics of laser-manipulated bromotrifluoromethane studied by x-ray absorption.
We present a computational study of the rotational molecular dynamics of bromotrifluoromethane (CF(3)Br) molecules in gas phase. The rotation is manipulated with an off-resonant 800 nm laser. The molecules are treated as rigid rotors. Frequently, we use a computationally efficient linear rotor model for CF(3)Br, which we compare with selected results for full symmetric-rotor computations. The expectation value (cos(2) theta)(t) is discussed. Especially, the transition from impulsive to adiabatic alignment, the temperature dependence of the maximally achievable alignment, and its intensity dependence are investigated. In a next step, we examine resonant x-ray absorption as an accurate tool to study laser manipulation of molecular rotation. Specifically, we investigate the impact of the x-ray pulse duration on the signal (particularly its temporal resolution) and study the temperature dependence of the achievable absorption. Most importantly, we demonstrated that using picosecond x-ray pulses, one can accurately measure the expectation value (cos(2) theta)(t) for impulsively aligned CF(3)Br molecules. We point out that a control of the rotational dynamics opens up a novel way to imprint shapes onto long x-ray pulses on a picosecond time scale. For our computations, we determine the dynamic polarizability tensor of CF(3)Br using ab initio molecular linear-response theory in conjunction with wave function models of increasing sophistication: Coupled-cluster singles (CCS), second-order approximate coupled-cluster singles and doubles (CC2), and coupled-cluster singles and doubles (CCSD).


Effect of oxygen on viability and substrate utilization in Chromatium.
Chromatium D can be exposed to oxygen for prolonged periods without any loss in motility or viability. Oxygen did not affect the rate of thiosulfate disappearance from the media, the oxidation of the inner sulfur atom of thiosulfate to sulfate, or the conversion of the outer sulfur atom of thiosulfate to intracellular sulfur, but it did inhibit the oxidation of intracellular sulfur to sulfate. Oxygen partially inhibited the uptake of pyruvate from the medium, but had little effect on the uptake of acetate. The distribution of label from pyruvate-2-(14)C into various cell fractions under aerobic conditions differed only slightly from that obtained under anaerobic conditions. Cells utilizing acetate-2-(14)C aerobically converted the majority of the metabolized acetate into a cell fraction with the solubility characteristics of poly-beta-hydroxybutyric acid, whereas under anaerobic conditions the acetate was distributed throughout the other cell fractions. Oxygen completely prevented the synthesis of bacteriochlorophyll.


A sculpting effect of reading on later representational quality of phonology revealed by multi-voxel pattern analysis in young children.
Using univariate analysis, a previous study by Wang et al. (2020) found a scaffolding effect of earlier phonological representation in superior temporal gyrus (STG) on later reading skill but failed to observe a sculpting effect of earlier reading on later phonological representation. The current study applied multi-voxel pattern analysis (MVPA) to examine if both scaffolding and sculpting effects were present in young children. We found that better initial reading skill predicted higher decoding coefficient of brain activity patterns for phonological representations in STG. This sculpting effect was present only for decoding small grain sizes (phonemes) and in younger children (6- to 7.5-year-olds), as we did not find any effects for large grain sizes (rhymes) or older children (7.5- to 9.5-year-olds). Although a scaffolding effect was not observed, the current study provides the first neural evidence of how earlier reading sculpts later phonological awareness in beginning readers.