Testosterone replacement therapy: improved sexual desire and erectile function in men with type 2 diabetes following a 30-week randomized placebo-controlled study.
Although testosterone replacement treatment (TRT) can improve sexual function in many hypogonadal (HG) men with type 2 diabetes (T2DM), some show either no improvement or only in a limited number of domains. Indeed, it is often difficult for the clinician to offer an indication of the likely efficacy of TRT as little data exist on the proportion of TRT-treated men who will demonstrate improvement in domains such as sexual desire (SxD) and erectile function (EF). We describe in men with T2DM: firstly, the likelihood of improved sexual desire (SxD) and erectile function (EF) following TRT at various time points, and secondly, if probability of SxD change predicted likelihood of subsequent EF change. During a 30-week randomized controlled study of testosterone undecanoate (TU), 199 T2DM men with HG (189 men completing) identified from primary care registers (placebo (P): 107, TU: 92) were stratified using baseline total testosterone (TT)/free testosterone (FT) into Mild (TT 8.1-12 nmol/L or FT 0.18-0.25 nmol/L) and Severe HG groups (TT ≤8 nmol/L and FT ≤0.18 nmol/L) and placebo (P)- and TU-treated groups. Associations between TU, SxD and EF were investigated using chi-square and logistic regression analysis. The proportion of men with improved SxD after 6 weeks and EF improvement after 30 weeks was significantly higher following TU treatment compared to P, this particularly evident in Severe HG men. Changes in SxD and EF were significantly associated in all groups. Logistic regression showed that SxD change at 6 weeks predicted of EF change after 30 weeks. Our study confirms TRT leads to changes in SxD and EF at different time points and suggests SxD and EF changes are related. SxD change after 6 weeks predicting EF change at 30 weeks is possibly a useful clinical finding.


[Clinical study of the cerebrovascular action of pentoxifylline].
Clinical trials of pentoxiphylline (trental) used in the treatment of 164 patients were carried out. Of these patients, 32 were suffering from acute cerebral circulation disorders, 25 from chronic cerebrovascular insufficiency, 50 from cerebrovascular dystonia of various genesis, 12 from peripheral vasculopathy, and 5 from migraine. A favourable effect of pentoxiphylline on the course of the cerebrovascular disorders (except migraine) was noted. Rheographic examinations confirmed the improvement of the cerebral and peripheral circulation and revealed a two-phase effect of the drug on the vascular tone that manifested in alternations of periods of short-time moderate vasodilatation and vasoconstriction. Pentoxiphylline was also found to produce a venotonic effect. The pharmacodynamic mechanisms of the hemovascular effects of the drug are discussed. The optimal dose of pentoxyìphylline for a single intravenous injection is assumed to be 10 ml of solution containing 200 mg of the drug dissolved in physiological saline. The optimal dose for drop infusion is up to 15--20 ml of the same solution. In addition the drug is to be given orally in a dose of 400 mg (two dragees of 200 mg) two or three times a day. The treatment lasts 3 to 5 weeks.


Tracking Gold Nanorods' Interaction with Large 3D Pancreatic-Stromal Tumor Spheroids by Multimodal Imaging: Fluorescence, Photoacoustic, and Photothermal Microscopies.
Pancreatic cancer is one of the most complex types of cancers to detect, diagnose, and treat. However, the field of nanomedicine has strong potential to address such challenges. When evaluating the diffusion and penetration of theranostic nanoparticles, the extracellular matrix (ECM) is of crucial importance because it acts as a barrier to the tumor microenvironment. In the present study, the penetration of functionalized, fluorescent gold nanorods into large (>500 μm) multicellular 3D tissue spheroids was studied using a multimodal imaging approach. The spheroids were generated by co-culturing pancreatic cancer cells and pancreatic stellate cells in multiple ratios to mimic variable tumor-stromal compositions and to investigate nanoparticle penetration. Fluorescence live imaging, photothermal, and photoacoustic analysis were utilized to examine nanoparticle behavior in the spheroids. Uniquely, the nanorods are intrinsically photoacoustic and photothermal, enabling multi-imaging detection even when fluorescence tracking is not possible or ideal.


Bean and Pea Plastoglobules Change in Response to Chilling Stress.
Plastoglobules (PGs) might be characterised as microdomains of the thylakoid membrane that serve as a platform to recruit proteins and metabolites in their spatial proximity in order to facilitate metabolic channelling or signal transduction. This study provides new insight into changes in PGs isolated from two plant species with different responses to chilling stress, namely chilling-tolerant pea (Pisum sativum) and chilling-sensitive bean (Phaseolus coccineus). Using multiple analytical methods, such as high-performance liquid chromatography and visualisation techniques including transmission electron microscopy and atomic force microscopy, we determined changes in PGs' biochemical and biophysical characteristics as a function of chilling stress. Some of the observed alterations occurred in both studied plant species, such as increased particle size and plastoquinone-9 content, while others were more typical of a particular type of response to chilling stress. Additionally, PGs of first green leaves were examined to highlight differences at this stage of development. Observed changes appear to be a dynamic response to the demands of photosynthetic membranes under stress conditions.


Elasticity of the supraspinatus tendon-muscle unit is preserved after acute tendon tearing in the rabbit.
Supraspinatus tendon tearing is one of the most common causes of the shoulder pain and dysfunction, which often requires a surgical repair. In this situation, proximal tendon stump is usually retracted medially from its original insertion. For successful reduction of the retracted tendon stump to its original insertion, the elasticity of the tendon-muscle unit should be preserved by the time of surgery. The purpose of the present study was to clarify the chronological changes in the elasticity of the supraspinatus tendon-muscle unit after acute tendon tearing to determine the optimal timing for the surgery. Right supraspinatus tendon was detached (detached side) in 40 male Japanese white rabbits, with left shoulders served as controls (control side). Eight animals were euthanized at 3 days and 1, 2, 4, or 8 weeks after surgery. Tissue sound speed that closely correlates to its elasticity was measured with a scanning acoustic microscope. In the supraspinatus tendon, tissue sound speed at 3 days after surgery was 1691.1 m/s, compared to 1714.3 m/s at the control side, but the difference was not statistically significant at any postoperative time period up to 8 weeks. In the supraspinatus muscle, tissue sound speed was not affected at all by the detachment of the tendon. The present study indicated that the elasticity of the supraspinatus tendon-muscle unit was well preserved for 8 weeks after the detachment. In the clinical practice, the retracted supraspinatus tendon stump could be repaired without excessive tension by 8 weeks from the acute tendon tearing.


Maternal MTHFR genotype and haplotype predict deficits in early cognitive development in a lead-exposed birth cohort in Mexico City.
Maternal folate nutritional status and prenatal lead exposure can influence fetal development and subsequent health. The methylenetetrahydrofolate reductase (MTHFR) gene is important for folate metabolism, and 2 common polymorphisms, C677T and A1298C, reduce enzymatic activity; C677T is present at high penetrance in Mexican populations. The objective of this study was to examine potential links between maternal and child MTHFR polymorphisms and child neurodevelopment in a lead-exposed population. Data regarding MTHFR polymorphisms C677T and A1298C, peri- and postnatal lead measures, and Bayley Mental Development Index at 24 mo of age (MDI-24) scores were available for 255 mother-child pairs who participated in the ELEMENT (Early Life Exposures in Mexico to Environmental Toxicants) study during 1994-1995. In covariate-adjusted regression models, maternal MTHFR 677 genotype predicted MDI-24 scores, in which each copy of the maternal MTHFR 677T variant allele was associated with lower MDI-24 scores (beta = -3.52; 95% CI: -6.12, -0.93; P = 0.004). Maternal MTHFR haplotype also predicted MDI-24 scores (mean +/- SE: 93.3 +/- 1.2 for 677C-1298A compared with 89.9 +/- 0.8 for 677T-1298A; P < 0.05). MDI-24 scores were not associated with maternal MTHFR 1298 genotype or child MTHFR genotypes. We did not observe significant MTHFR genotype x lead interactions with respect to any of the subject biomarkers of lead exposure. The maternal MTHFR 677T allele is an independent predictor of poorer child neurodevelopment at 24 mo. These results suggest that maternal genetic variations in folate metabolism during pregnancy may program offspring neurodevelopment trajectories. Further research is warranted to determine the generalizability of these results across other populations.


Multiple outbreaks of acute hemorrhagic conjunctivitis due to a variant of coxsackievirus A24: Guangdong, China, 2007.
Acute hemorrhagic conjunctivitis (AHC) is usually caused by enterovirus 70, coxsackievirus A24(CA24v) and adenoviruses. Several outbreaks of AHC caused by a CA24v have occurred since it was imported into China in 1971. Multiple outbreaks of AHC reappeared in 10 cities of Guangdong during June to November in 2007. The epidemic began in the June, and spread extensively, with a peak in the September. A total of 31,659 cases were reported to center for disease control and prevention of Guangdong, it was estimated that the number of actual AHC was >200 thousands. Forty conjunctival swab specimens were collected from the cases diagnosed clinically with AHC. (RT)-PCR testing on these conjunctival specimens revealed the presence of an enterovirus, and this was confirmed by 16 isolates. We demonstrated the most likely etiological agent for the multiple outbreaks was a variant of coxsackievirus A24 by molecular typing using a partial VP1 sequence. Sequence comparison and phylogenetic analyses of the VP1 and 3Cpro gene regions were performed by Neighbor-joining method, the strains from different outbreaks and different geographical areas within Guangdong had no sequence divergence in 2007. The representative isolates from mainland of China including Hangzhou, Ningbo, Beijing, Yunnan, Liaoning, and Henan were analyzed in this study. Phylogenetic analysis revealed theses isolates were located in different clusters, a close phylogenetic and chronological relationship with Singaporean, South Korean and Thailand isolates had been observed. This confirms CA24v circulated in China's mainland has not evolved independently, but co-evolved with the isolates of Southeast Asia.


Learning Local Descriptors by Optimizing the Keypoint-Correspondence Criterion: Applications to Face Matching, Learning From Unlabeled Videos and 3D-Shape Retrieval.
Current best local descriptors are learned on a large data set of matching and non-matching keypoint pairs. However, data of this kind are not always available, since the detailed keypoint correspondences can be hard to establish. On the other hand, we can often obtain labels for pairs of keypoint bags. For example, keypoint bags extracted from two images of the same object under different views form a matching pair, and keypoint bags extracted from images of different objects form a non-matching pair. On average, matching pairs should contain more corresponding keypoints than non-matching pairs. We describe an end-to-end differentiable architecture that enables the learning of local keypoint descriptors from such weakly labeled data. In addition, we discuss how to improve the method by incorporating the procedure of mining hard negatives. We also show how our approach can be used to learn convolutional features from unlabeled video signals and 3D models.


Successful use of intra-aortic counter pulsation therapy for intractable ventricular arrhythmia in patient with severe left ventricular dysfunction and normal coronary arteries.
Intra-aortic balloon pumps (IABP) are commonly used in the setting of an acute myocardial infarction that is complicated by cardiogenic shock or mechanical complications such as a ventricular septal defect or papillary muscle rupture. IABP has also been shown to be useful in patients with refractory and hemodynamically unstable ventricular arrhythmias and refractory post-myocardial infarction angina. We report a case in which IABP was used in a patient with dilated cardiomyopathy and normal coronary arteries, who presented with persistent, recurrent and refractory ventricular tachycardia. His ventricular tachycardia settled immediately with the use of IABP therapy. He subsequently had an implantable defibrillator. The use of IABP is associated with favorable changes in the left ventricular wall tension and reduction in afterload, which could reduce the excitability of the myocardium, thus making it less prone to arrhythmias. The use of IABP is relatively safe and should be considered in patients with refractory ventricular arrhythmias, even if it is not associated with ischemia.


Increases in DNA lesions and the DNA damage indicator Gadd45 following transient cerebral ischemia.
Transient global or focal ischemia leads to the production of several types of lesions in the DNA backbone including alkali-labile sites, and both single-stranded (ss) and double-stranded (ds) breaks. The ds breaks result in high molecular weight fragments of 10-50 kbp that contain both 3'- and 5'-OH end-groups, suggesting that more than one endonuclease is involved. This lesioning of DNA is followed by the appearance of the damage-response indicator Gadd45 in the ischemic hemisphere following middle cerebral artery occlusion. By 6 h, gadd45 mRNA was shown to increase by semi-quantitative reverse transcriptase - polymerase chain reaction. In situ hybridization histochemistry indicated that these increases in gadd45 mRNA occurred in pyramidal neurons located on the edge of the infarcted cortex. Gadd45 immunostaining yielded similar findings with maximal protein staining detected at 18 h after occlusion. In neurons, in the infarct core with frank DNA fragmentation shown by in situ TdT-mediated dUTP-biotin nick end labeling (TUNEL) at 24 h, the Gadd45 immunostaining was not visible. Taken together, these findings suggest that Gadd45 responds to DNA damage following ischemia as part of a repair response mounted by brain cells attempting to survive the insult.


Relationship of Indoor, Outdoor and Personal Air (RIOPA) study: study design, methods and quality assurance/control results.
The Relationship of Indoor, Outdoor and Personal Air (RIOPA) Study was undertaken to evaluate the contribution of outdoor sources of air toxics, as defined in the 1990 Clean Air Act Amendments, to indoor concentrations and personal exposures. The concentrations of 18 volatile organic compounds (VOCs), 17 carbonyl compounds, and fine particulate matter mass (PM(2.5)) were measured using 48-h outdoor, indoor and personal air samples collected simultaneously. PM2.5 mass, as well as several component species (elemental carbon, organic carbon, polyaromatic hydrocarbons and elemental analysis) were also measured; only PM(2.5) mass is reported here. Questionnaires were administered to characterize homes, neighborhoods and personal activities that might affect exposures. The air exchange rate was also measured in each home. Homes in close proximity (<0.5 km) to sources of air toxics were preferentially (2:1) selected for sampling. Approximately 100 non-smoking households in each of Elizabeth, NJ, Houston, TX, and Los Angeles, CA were sampled (100, 105, and 105 respectively) with second visits performed at 84, 93, and 81 homes in each city, respectively. VOC samples were collected at all homes, carbonyls at 90% and PM(2.5) at 60% of the homes. Personal samples were collected from nonsmoking adults and a portion of children living in the target homes. This manuscript provides the RIOPA study design and quality control and assurance data. The results from the RIOPA study can potentially provide information on the influence of ambient sources on indoor air concentrations and exposure for many air toxics and will furnish an opportunity to evaluate exposure models for these compounds.


Pathophysiology of cerebral oedema in acute liver failure.
Cerebral oedema is a devastating consequence of acute liver failure (ALF) and may be associated with the development of intracranial hypertension and death. In ALF, some patients may develop cerebral oedema and increased intracranial pressure but progression to life-threatening intracranial hypertension is less frequent than previously described, complicating less than one third of cases who have proceeded to coma since the advent of improved clinical care. The rapid onset of encephalopathy may be dramatic with the development of asterixis, delirium, seizures and coma. Cytotoxic and vasogenic oedema mechanisms have been implicated with a preponderance of experimental data favouring a cytotoxic mechanism. Astrocyte swelling is the most consistent neuropathological finding in humans with ALF and ammonia plays a definitive role in the development of cytotoxic brain oedema. The mechanism(s) by which ammonia induces astrocyte swelling remains unclear but glutamine accumulation within astrocytes has led to the osmolyte hypothesis. Current evidence also supports an alternate 'Trojan horse' hypothesis, with glutamine as a carrier of ammonia into mitochondria, where its accumulation results in oxidative stress, energy failure and ultimately astrocyte swelling. Although a complete breakdown of the blood-brain barrier is not evident in human ALF, increased permeation to water and other small molecules such as ammonia has been demonstrated resulting from subtle alterations in the protein composition of paracellular tight junctions. At present, there is no fully efficacious therapy for cerebral oedema other than liver transplantation and this reflects our incomplete knowledge of the precise mechanisms underlying this process which remain largely unknown.


[A retrospective clinical analysis of 16 patients with scleroderma renal crisis].
To analyze the clinical features, laboratory tests, treatments and outcome of patients with scleroderma renal crisis (SRC). We retrospectively reviewed the clinical and laboratory data of 16 patients with scleroderma renal crisis in Peking Union Medical College Hospital from May 2004 to May 2013. The treatment and outcome of SRC patients were also retrospectively analyzed. There were a total of 16 SRC patients including 5 male patients and 11 females. The median age at SRC onset was (49.9 ± 12.3) years. It usually took 3.2 years from the diagnosis of systemic sclerosis (SSc) to SRC attack. Ten SRC patients belonged to diffuse cutaneous systemic sclerosis (dcSSc), and 6 patients were limited cutaneous systemic sclerosis (lcSSc). Among SRC patients, 16/16 were negative of anti-centromere antibodies (ACAs). All these 16 patients had hypertension and renal insufficiency, including 8 dialysis dependent after the onset of SRC and 7 with thrombotic microangiopathy. There were 3 patients receiving renal biopsy. The pathological findings were mainly summarized as intimal thickening and stenosis of renal arterioles. Among 13 patients with long-term followed-up, 11 patients received angiotensin converting enzyme inhibitors (ACEI), 5 patients died, 2 patients were dialysis dependent. Only 1 patient stopped dialysis after the combination treatment of ACEI and endothelin receptor antagonist. Another 5 patients didn't need dialysis. SRC usually occurred at the early course of SSc. dcSSc was more frequent than lcSSc. ACAs were rarely found in SRC patients. The immediate and sufficient use of ACEIs was still the cornerstone of SRC treatment. Future studies are needed to evaluate the efficacy of endothelin receptor antagonist in the treatment of SRC.


Caring for hemodialysis patients in Saudi Arabia. Past, present and future.
There are currently 5706 patients receiving hemodialysis therapy in the Kingdom of Saudi Arabia - a 15 fold increase when compared to 1983. The annual increase in the number of patients on dialysis for 1999 is 696 (10 fold increase when compared to 1983). Besides the massive increase in the number of patients in the last 20 years, we have noticed a marked increase in the mean age of patients (51.3 years in 1999 as compared to 37.9 years in the early 80s). Diabetes mellitus which was an insignificant contributory etiology (4%) in the early 80s is now a major cause (16-25%). Similarly mortality has increased from 4% annually to 11-14% annually. This is largely due to increasing age and prevalence of diabetes mellitus. Within the expired cohort the mean age was 62.3 years compared to 51.3 years of the total dialysis population, and diabetes mellitus was present in 60.5% in those who expired. Moreover, ischemic heart disease was diagnosed in 50% before death. Tuberculosis and Hepatitis C virus incidences, however, have not improved over the years but the degree of rehabilitation has, largely due to better hemoglobin level and due to the technological advances in dialysis delivery. This article describes these changes, their causes and implications.


Striking Elevations in Aminotransferases in a Case of Congestive Hepatopathy Without Concurrent Hypotension.
Congestive hepatopathy results from passive venous congestion often in the setting of right heart failure. Injury to the liver due to congestion is often asymptomatic and may be difficult to recognize and diagnose. The degree of elevations in aminotransferases varies in cases of congestive hepatopathy but usually stays within two to three times the upper limit of normal. Here, we report an interesting case of congestive hepatopathy that presented with striking elevations of aminotransferases in the 2000s international units/liter a few days after admission without concurrent hypotension.


Antinociceptive effects of monoamine reuptake inhibitors administered alone or in combination with mu opioid agonists in rhesus monkeys.
Cocaine, which non-selectively blocks the reuptake of the monoamines serotonin, dopamine and norepinephrine, produces weak antinociceptive effects and increases the antinociceptive effects of low- to intermediate-efficacy mu opioid agonists in rhesus monkeys. In the present study, the antinociceptive effects of more selective monoamine reuptake inhibitors administered alone and in combination with mu opioid agonists were evaluated in rhesus monkeys using a warm-water tail-withdrawal assay of thermal nociception. Like cocaine, the selective serotonin reuptake inhibitors clomipramine (0.01-3.2 mg/kg) and fluoxetine (0.1-10 mg/kg) produced weak antinociceptive effects. Pretreatment with the serotonin receptor antagonist mianserin (0.032-0.32 mg/kg) produced rightward and downward shifts in the clomipramine dose-effect curve, suggesting that the effects of clomipramine were mediated by serotonin receptors. Combination of clomipramine with the low efficacy mu agonist nalbuphine or the intermediate efficacy mu agonist morphine produced more antinociception than did the mu agonists alone. Fluoxetine also produced a small leftward shift in the morphine dose-effect curve. The selective norepinephrine reuptake inhibitors nisoxetine (0.1-10 mg/kg) and tomoxetine (0.1-10 mg/kg) and the selective dopamine reuptake inhibitors bupropion (0.032-3.2 mg/kg) and GBR 12909 (0.1-10 mg/kg) did not produce antinociception or increase antinociception induced by nalbuphine or morphine. In fact, GBR 12909 produced dose-dependent allodynia and reduced the maximal antinociceptive effects of morphine. These results suggest that inhibition of serotonin reuptake is sufficient to produce weak antinociceptive effects and enhance the antinociceptive effects of low efficacy mu opioid agonists. These results also suggest that the effects of cocaine on serotonin reuptake may contribute to cocaine's antinociceptive effects in rhesus monkeys.


Patient and Clinician Characteristics Associated With Secure Message Content: Retrospective Cohort Study.
Good communication has been shown to affect patient outcomes; however, the effect varies according to patient and clinician characteristics. To date, no research has explored the differences in the content of secure messages based on these characteristics. This study aims to explore characteristics of patients and clinic staff associated with the content exchanged in secure messages. We coded 18,309 messages that were part of threads initiated by 1031 patients with hypertension, diabetes, or both conditions, in communication with 711 staff members. We conducted four sets of analyses to identify associations between patient characteristics and the types of messages they sent, staff characteristics and the types of messages they sent, staff characteristics and the types of messages patients sent to them, and patient characteristics and the types of messages they received from staff. Logistic regression was used to estimate the strength of the associations. We found that younger patients had reduced odds of sharing clinical updates (odds ratio [OR] 0.77, 95% CI 0.65-0.91) and requesting prescription refills (OR 0.77, 95% CI 0.65-0.90). Women had reduced odds of self-reporting biometrics (OR 0.78, 95% CI 0.62-0.98) but greater odds of responding to a clinician (OR 1.20, 95% CI 1.02-1.42) and seeking medical guidance (OR 1.19, 95% CI 1.01-1.40). Compared with White patients, Black patients had greater odds of requesting preventive care (OR 2.68, 95% CI 1.30-5.51) but reduced odds of requesting a new or changed prescription (OR 0.72, 95% CI 0.53-0.98) or laboratory or other diagnostic procedures (OR 0.66, 95% CI 0.46-0.95). Staff had lower odds of sharing medical guidance with younger patients (OR 0.83, 95% CI 0.69-1.00) and uninsured patients (OR 0.21, 95% CI 0.06-0.73) but had greater odds of sharing medical guidance with patients with public payers (OR 2.03, 95% CI 1.26-3.25) compared with patients with private payers. Staff had reduced odds of confirming to women that their requests were fulfilled (OR 0.82, 95% CI 0.69-0.98). Compared with physicians, nurse practitioners had greater odds of sharing medical guidance with patients (OR 2.74, 95% CI 1.12-6.68) and receiving prescription refill requests (OR 3.39, 95% CI 1.49-7.71). Registered nurses had greater odds of deferred information sharing (OR 1.61, 95% CI 1.04-2.49) and receiving responses to messages (OR 3.93, 95% CI 2.18-7.11) than physicians. The differences we found in content use based on patient characteristics could lead to the exacerbation of health disparities when content is associated with health outcomes. Disparities in the content of secure messages could exacerbate disparities in patient outcomes, such as satisfaction, trust in the system, self-care, and health outcomes. Staff and administrators should evaluate how secure messaging is used to ensure that disparities in care are not perpetuated via this communication modality.


[Proliferative capacity of mesenchymal stem cells from human fetal bone marrow and their ability to differentiate into the derivative cell types of three embryonic germ layers].
Strong proliferative capacity and the ability to differentiate into the derivative cell types of three embryonic germ layers are the two important characteristics of embryonic stem cells. To study whether the mesenchymal stem cells from human fetal bone marrow (hfBM-MSCs) possess these embryonic stem cell-like biological characteristics, hfBM-MSCs were isolated from bone barrows and further purified according to the different adherence of different kinds of cells to the wall of culture flask. The cell cycle of hfBM-MSCs and MSC-specific surface markers such as CD29, CD44, etc were identified using flow cytometry. The expressions of human telomerase reverse transcriptase (hTERT), the embryonic stem cell-specific antigens, such as Oct4 and SSEA-4 were detected with immunocytochemistry at the protein level and were also tested by RT-PCR at the mRNA level. Then, hfBM-MSCs were induced to differentiate toward neuron cells, adipose cells, and islet B cells under certain conditions. It was found that 92.3% passage-4 hfBM-MSCs and 96.1% passage-5 hfBM-MSCs were at G(0)/G(1) phase respectively. hfBM-MSCs expressed CD44, CD106 and adhesion molecule CD29, but not antigens of hematopoietic cells CD34 and CD45, and almost not antigens related to graft-versus-host disease (GVHD), such as HLA-DR, CD40 and CD80. hfBM-MSCs expressed the embryonic stem cell-specific antigens such as Oct4, SSEA-4, and also hTERT. Exposure of these cells to various inductive agents resulted in morphological changes towards neuron-like cells, adipose-like cells, and islet B-like cells and they were tested to be positive for related characteristic markers. These results suggest that there are plenty of MSCs in human fetal bone marrow, and hfBM-MSCs possess the embryonic stem cell-like biological characteristics, moreover, they have a lower immunogenic nature. Thus, hfBM-MSCs provide an ideal source for tissue engineering and cellular therapeutics.


Towards inclusive notions of 'patient' and 'nurse'.
Nursing is commonly characterized by the process of caring in the context of nurse-patient interactions in clinical and non-clinical settings. This interactive context of nursing however should not be seen to only involve encounters of an interpersonal nature but also must be extended to include interactions set in a social-relational context. (The growing realization that most health problems in our society cannot be fully understood as only an individual phenomena lends support to this notion.) Continuity and quality of time are described as being important aspects of nursing, which make possible a holistic and unified approach to care that is quite unique to nursing. A patient-centred approach to nursing is contrasted with an illness/disease-based approach. Growing recognition of the positive outcomes of an approach to nursing which is responsive to patients (rather than primarily focused on illness/disease) suggests that patterns of nursing care should be more closely aligned with this model. The authors believe this can be accomplished through a supportive emotional atmosphere of interpersonal and social-relational involvement. The modes by which nurses will be accountable to the community will then necessarily need to reflect this broader, more inclusive context.


Potential effects of plant protease inhibitors, oryzacystatin I and soybean Bowman-Birk inhibitor, on the aphid parasitoid Aphidius ervi Haliday (Hymenoptera, Braconidae).
Protease inhibitors (PIs) have been shown to cause lethal and sublethal effects on aphids depending on the kind of PI and aphid species. Therefore, these proteins might affect aphid parasitoids directly by inhibiting their digestive proteolysis or indirectly via their development in a less suitable host. In our study, the risk of exposure and the potential effects of soybean Bowman-Birk inhibitor (SbBBI) and oryzacystatin I (OCI) on the aphid endoparasitoid Aphidius ervi were investigated using artificial diet to deliver PIs. Immunoassays showed that both SbBBI and OCI were detected in the honeydew of aphids reared on artificial diet containing these recombinant proteins at 100 microg/mL. However, only SbBBI was detected in parasitoid larvae, while this PI could not be detected in adult parasitoids emerged from PI-intoxicated aphids. Enzymatic inhibition assays showed that digestive proteolytic activity of larvae and adults of A. ervi predominantly relies on serine proteases and especially on chymotrypsin-like activity. Bioassays using SbBBI and OCI on artificial diet were performed. A. ervi that developed on intoxicated aphids had impaired fitness. Thus development and parasitism success of parasitoids exposed to OCI were severely affected. On the contrary, SbBBI only altered significantly female size and sex ratio. Direct exposure to PIs through adult food intake did not affect female's longevity, while SbBBI and OCI (100 microg/mL) induced 69% and 30% inhibition of digestive protease activity, respectively. These studies made it possible to estimate the risk of exposure to plant PIs and the sensitivity of the aphid parasitoid A. ervi to these entomotoxins, by combining immunological, biochemical and biological approaches. First it pointed out that only immature stages are affected by PIs. Secondly, it documented two different modes of effect, according to the nature of the PIs and both host and parasitoid susceptibility. OCI prevented the development of A. ervi mainly due to the host susceptibility, whereas SbBBI only induced sublethal effects on the parasitoid, possibly due to both direct action on the parasitoid susceptible proteases, and host-mediated action through size reduction.


Professional carers' knowledge and response to depression among their aged-care clients: the care recipients' perspective.
Depression is an under-diagnosed disorder among the elderly, even in those who are in receipt of aged-care services. One factor associated with this under diagnosis has been identified as a reluctance amongst the elderly to discuss their mood and emotions with their medical practitioners. The current study focused on why depression is not recognised and acted on by those providing residential or home-based care to older people. We interviewed 15 elderly people residing in high-level or low-level aged-care facilities, and three elderly people who were receiving personal care in their homes. All participants had been identified by their care agencies as depressed. Participants reported their perceptions of their personal carers' knowledge and practices in managing the residents' depression. Although the participants described their carers in positive terms, they were critical of their knowledge and skills in recognising depression, and indicated that the communication between personal carers and care recipients about depressive symptomatology was seriously flawed. Training for personal carers in these areas, and efforts to change organisational culture are recommended.


Vaccination with post-translational modified, homocitrullinated peptides induces CD8 T-cell responses that mediate antitumor immunity.
Post-translational modification of proteins has the potential to alter the ability of T cells to recognize major histocompatibility complex (MHC) class -I and class-II restricted antigens, thereby resulting in altered immune responses. One such modification is carbamylation (homocitrullination) that results in the formation of homocitrulline (Hcit) residues in a non-enzymatic reaction of cyanate with the lysine residues in the polypeptide chain. Homocitrullination occurs in the tumor microenvironment and CD4-mediated immune responses to Hcit epitopes can target stressed tumor cells and provide a potent antitumor response in mouse models. Homocitrullinated peptides were identified and assessed in vitro for HLA-A2 binding and in vivo in human leukocyte antigen (HLA) transgenic mouse models for immunogenicity. CD8 responses were assessed in vitro for cytotoxicity and in vivo tumor therapy. Human tumor samples were analyzed by targeted mass spectrometry for presence of homocitrullinated peptides. Homocitrullinated peptides from aldolase and cytokeratin were identified, that stimulated CD8-mediated responses in vivo. Modified peptides showed enhanced binding to HLA-A2 compared with the native sequences and immunization of HLA-A2 transgenic mice generated high avidity modification specific CD8 responses that killed peptide expressing target cells. Importantly, in vivo the homocitrullinated aldolase specific response was associated with efficient CD8 dependent antitumor therapy of the aggressive murine B16 tumor model indicating that this epitope is naturally presented in the tumor. In addition, the homocitrullinated aldolase epitope was also detected in human tumor samples. This is the first evidence that homocitrullinated peptides can be processed and presented via MHC-I and targeted for tumor therapy. Thus, Hcit-specific CD8 T-cell responses have potential in the development of future anticancer therapy.


Assessments of variability in cortical and subcortical measurements and within-network connectivity of the brain using test-retest data.
The evaluation and diagnosis of structural changes in brain caused by disease or treatment over time has become one of the important applications of medical imaging methods, in particular MRI, and it is growing. It is critical to evaluate the reliability of the changes in measurements observed in an individual patient for any clinical decision-making. In this paper, we calculated the repeatability coefficient (RC) as a measure of uncertainty for MRI measurements of subcortical volumes and cortical thickness, and within-network connectivity using test-retest data of 20 healthy subjects. We also evaluated changes in 13 patients who received 20 sessions of transcranial magnetic stimulation as a treatment. The most reliable measure seems to be in the thickness of the left occipital with RC% of 3.5 and the least reliable measure is the brain connectivity within visual network using Yeo's atlas with RC% of 29.4. The most sensitive measure to the percentage of true changes in treated patients is the connectivity within subcortical network of AAL with 76.9%.Clinical Relevance- The results of this study can be useful for evaluating changes in the gray matter structures or functional connectivity of the brain due to a neurological disease such as Alzheimer's or Parkinson's. Also, the obtained results can be used to evaluate the changes caused by any intervention or treatment that may have any positive or negative effect on the brain.


Involvement of Src tyrosine kinases (SFKs) and of focal adhesion kinase (FAK) in the injurious mechanism in rat primary neuronal cultures exposed to chemical ischemia.
Src family of kinases (SFKs) and focal adhesion kinase (FAK) are two important cellular signaling components known to act cooperatively in the transduction of death and survival signals. We investigated the involvement of these proteins in the mechanism of the injurious response in rat primary neuronal cultures exposed to an insult composed of chemical ischemia (poisoning with iodoacetic acid; 100 muM, for 150 min) followed by 1 h of incubation in the regular medium, an insult shown before to be associated with generation of reactive oxygen species and with the depletion of adenosine triphisphate. The exposure of the neuronal cultures to the insult resulted in cell injury, assessed by the increased release of cytoplasmic lactate dehydrogenase (LDH) into the culture media, which could be attenuated markedly by the presence of the antioxidant LY 231617. The insult resulted in the decreased level of phosphorylation of the SFKs members Src, Fyn, and Yes at the Src Y416-equivalent activation sites and of the FAK Y397 activation site, degradation of FAK to a p85 fragment, and disassembling of the FAK-SFKs complexes. The inhibition of SFKs was found to be responsible for part of the insult-induced cell damage manifested in increased LDH release. Pervanadate, an inhibitor of the phosphotyrosine phosphatases (PTPs), abrogated the inactivation of SFKs and attenuated cell injury, indicating that insult-induced activation of PTPs is involved in SFKs inhibition and the ensued damage. The inhibition of SFKs and FAK is probably the cause of the disassembling of SFKs-FAK complexes, a process known to be associated with apoptosis.


Does cognition-specific computer training have better clinical outcomes than non-specific computer training? A single-blind, randomized controlled trial.
The purpose of this study was to investigate differences between non-specific computer training (NCT) and cognition-specific computer training (CCT). Randomized controlled experimental study. Local community welfare center. A total of 78 subjects with mild cognitive impairment (MCI) were randomly assigned to the NCT ( n = 39) or CCT group ( n = 39). The NCT group underwent NCT using Nintendo Wii for improving functional performance, while the CCT group underwent CCT using CoTras for improving function of the cognitive domain specifically. Subjects in both groups received 30-minute intervention three times a week for 10 weeks. To identify effects on cognitive function, the Wechsler Adult Intelligence Scale (WAIS) digit span subtests, Rey Auditory Verbal Learning Test (RAVLT), Trail Making Test-Part B (TMT-B), Rey-Osterrieth Complex Figure Test, and Modified Taylor Complex Figure (MTCF) were used. Health-related quality of life (HRQoL) was assessed using the Short-Form 36-item questionnaire. After 10 weeks, the WAIS subtests (digit span forward: 0.48 ± 0.08 vs. 0.12 ± 0.04; digit span backward: 0.46 ± 0.09 vs. 0.11 ± 0.04) and HRQoL (vitality: 9.05 ± 1.17 vs. 2.69 ± 1.67; role-emotional: 8.31 ± 1.20 vs. 4.15 ± 0.71; mental health: 11.62 ± 1.63 vs. 6.95 ± 1.75; bodily pain: 4.21 ± 2.17 vs. 0.10 ± 0.38) were significantly higher in the NCT group ( P < 0.05). NCT was superior to CCT for improving cognitive function and HRQoL of elderly adults with MCI.


Comparative Study of Molecular Basket Sorbents Consisting of Polyallylamine and Polyethylenimine Functionalized SBA-15 for CO2 Capture from Flue Gas.
Polyallylamine (PAA)-based molecular basket sorbents (MBS) have been studied for CO2 capture in comparison with polyethylenimine (PEI)-based MBS. The characterizations including N2 physisorption, diffuse reflectance infrared Fourier transform spectroscopy (DRIFTS), and thermogravimetric analysis (TGA) showed that PAA (Mn =15 000) is more rigid and has more steric hindrance inside SBA-15 pores than PEI owing mainly to its different polymer structure. The effects of temperature and PAA loading on the CO2 sorption capacity of PAA-based MBS have been examined by TGA by using 100 % CO2 gas stream and compared with PEI/SBA-15. It was found that the capacity of the PAA/SBA-15 sorbent increased with increasing temperature. The optimum capacity of 88 mgCO2 gsorb-1 was obtained at 140 °C for PAA(50)/SBA-15 whereas the optimum sorption temperature was 75 and 90 °C for PEI-I(50)/SBA-15 (PEI-I, Mn =423) and PEI-II(50)/SBA-15 (PEI-II, Mn =25 000), respectively. The capacity initially increased with the increase of PAA loading and then dropped at high amine contents, owing to the increased diffusion barrier. The highest CO2 capacity of 109 mgCO2 gsorb-1 was obtained at a PAA loading of 65 wt %, whereas the PAA(50)/SBA-15 sorbent gave the best amine efficiency of 0.23 molCO2 molN-1 . The effect of moisture was examined in a fixed-bed flow system with simulated flue gas containing 15 % CO2 and 4.5 % O2 in N2 . It was found that the presence of moisture significantly enhanced CO2 sorption over PAA(50)/SBA-15 and greatly improved its cyclic stability and regenerability. Compared with PEI/SBA-15, PAA/SBA-15 possesses a better thermal stability and higher resistance to oxidative degradation. However, the CO2 sorption rate over the PAA(50)/SBA-15 sorbent was much slower.


Full field tabletop EUV coherent diffractive imaging in a transmission geometry.
We demonstrate the first general tabletop EUV coherent microscope that can image extended, non-isolated, non-periodic, objects. By implementing keyhole coherent diffractive imaging with curved mirrors and a tabletop high harmonic source, we achieve improved efficiency of the imaging system as well as more uniform illumination at the sample, when compared with what is possible using Fresnel zone plates. Moreover, we show that the unscattered light from a semi-transparent sample can be used as a holographic reference wave, allowing quantitative information about the thickness of the sample to be extracted from the retrieved image. Finally, we show that excellent tabletop image fidelity is achieved by comparing the retrieved images with scanning electron and atomic force microscopy images, and show superior capabilities in some cases.


Cardioprotection of Eucommiae Folium: the extract and pharmaceutically active compounds attenuate hyperglycemia, mitochondrial dysfunction, calcium dyshomeostasis, insulin resistance, and oxidative stress in diabetic cardiomyopathy db/db mice.
A biolabel research based on multi-omics, informatics, molecular docking, and experimental verification was used to investigate the cardioprotective effect and pharmaceutically active compounds of Eucommiae Folium (EF). Based on the biolabel research pattern, metabonomics, proteomics, and bioinformatics indicated that EF has a therapeutic potential for a variety of heart diseases, especially cardiomyopathy, and the most critical mechanism involved is the diabetic cardiomyopathy pathway. Bioinformatics, cheminformatics, and molecular docking showed that 24 EF compounds may play a therapeutic role in diabetic cardiomyopathy via this pathway. Among which, four compounds (kaempferol, esculetin, (+)-catechin, and astragalin) showed appropriate pharmacokinetic parameters and formed stable binding with biolabels in the pathway. In diabetic cardiomyopathy db/db mice, histopathological analysis, mitochondrial swelling and membrane potential assay, ELISA, and biochemical analysis demonstrated that EF and four active compounds had obvious hypoglycemic effects and attenuated myocardial damage and related pathological processes, such as mitochondrial dysfunction, calcium dyshomeostasis, insulin resistance, and oxidative stress. This study provides new evidence and insights into the effect, mechanism, and material basis of EF in treating diabetic cardiomyopathy.


Resolving Chemical Modifications to a Single Amino Acid within a Peptide Using a Biological Nanopore.
While DNA sequencing is now amply available, fast, and inexpensive, protein sequencing remains a tremendous challenge. Nanopores may allow for developing a protein sequencer with single-molecule capabilities. As identification of 20 different amino acids currently presents an unsurmountable challenge, fingerprinting schemes are pursued, in which only a subset of amino acids is labeled and detected. This requires modification of amino acids with chemical structures that generate a distinct nanopore ionic current signal. Here, we use a model peptide and the fragaceatoxin C nanopore to characterize six potential tags for a fingerprinting approach using nanopores. We find that labeled and unlabeled proteins can be clearly distinguished and that sensitive detection is obtained for labels with a spectrum of different physicochemical properties such as mass (427-1275 Da), geometry, charge, and hydrophobicity. Additionally, information about the position of the label along the peptide chain can be obtained from individual current-blockade event features. The results represent an important advance toward the development of a single-molecule protein-fingerprinting device with nanopores.


Comparative analysis of DNA methylation patterns in transgenic Drosophila overexpressing mouse DNA methyltransferases.
DNA methyltransferases (Dnmts) mediate the epigenetic modification of eukaryotic genomes. Mammalian DNA methylation patterns are established and maintained by co-operative interactions among the Dnmt proteins Dnmt1, Dnmt3a and Dnmt3b. Owing to their simultaneous presence in mammalian cells, the activities of individual Dnmt have not yet been determined. This includes a fourth putative Dnmt, namely Dnmt2, which has failed to reveal any activity in previous assays. We have now established transgenic Drosophila strains that allow for individual overexpression of all known mouse Dnmts. Quantitative analysis of genomic cytosine methylation levels demonstrated a robust Dnmt activity for the de novo methyltransferases Dnmt3a and Dnmt3b. In addition, we also detected a weak but significant activity for Dnmt2. Subsequent methylation tract analysis by genomic bisulphite sequencing revealed that Dnmt3 enzymes preferentially methylated CpG dinucleotides in a processive manner, whereas Dnmt2 methylated isolated cytosine residues in a non-CpG dinucleotide context. Our results allow a direct comparison of the activities of mammalian Dnmts and suggest a significant functional specialization of these enzymes.


Reproducibility of DNA ploidy and S-phase values from paraffin-embedded tissue.
To determine the degree of variability of DNA flow cytometric results between two types of flow cytometers. Single-cell suspensions were made from 40 paraffin blocks of ovarian carcinoma using a modified Hedley procedure. These samples were consecutively analyzed in both Coulter's Profile II and Elite Flow cytometers. Gated and ungated data were collected and analyzed using Phoenix flow system's multicycle software. There were 16 (40%) DNA diploid, 23 (58%) DNA aneuploid and 1 DNA tetraploid tumor. The degree of variability in the DNA index coefficient of variation of the G0/G1 diploid peak, percentage of S phase, percentage of G2M, percentage of debris plus percentage of clumps were compared. Excellent correlations of the results were obtained in the DNA index (r = .999) and in percentage of S phase (r = .946). It is feasible to standardize variables of flow cytometric instruments to obtain reproducible results.


Inhibition of Excision of Oxidatively Generated Hydantoin DNA Lesions by NEIL1 by the Competitive Binding of the Nucleotide Excision Repair Factor XPC-RAD23B.
The interplay between nucleotide excision repair (NER) and base excision repair (BER) of nonbulky, oxidatively generated DNA lesions has long been a subject of significant interest. The hydantoin oxidation products of 8-oxoguanine, spiroiminodihydantoin (Sp) and 5-guanidinohydantoin (Gh), are substrates of both BER and NER in HeLa cell extracts and human cells [Shafirovich, V., et al. (2019) Chem. Res. Toxicol. 32, 753-761]. The primary factor that recognizes DNA lesions is the DNA damage-sensing factor XPC-RAD23B (XPC), while the glycosylase NEIL1 is known to remove Gh and Sp lesions from double-stranded DNA. It is shown here that in aqueous solutions containing nanomolar concentrations of proteins, XPC and NEIL1 compete for binding to 147-mer oligonucleotide duplexes that contain single Gh or Sp lesions under conditions of [protein] ≫ [DNA], thus inhibiting the rate of BER catalyzed by NEIL1. The non-covalently bound NEIL1 molecules can be displaced by XPC at concentration ratios R = [XPC]/[NEIL1] > 0.2, while full displacement of NEIL1 is observed at R ≥ 0.5. In the absence of XPC and under single-turnover conditions, only the burst phase is observable. However, with a progressive increase in the XPC concentration, the amplitude of the burst phase decreases gradually, and a slower time-dependent phase of incision product formation manifests itself with rate constants of 3.0 × 10-3 s-1 (Gh) and 0.90 × 10-3 s-1 (Sp). These slow kinetics are attributed to the dissociation of XPC-DNA complexes that allow for the rebinding of NEIL1 to the temporarily exposed Gh or Sp lesions, and the incisions observed under these steady-state conditions.


Distribution pattern of hepatitis C virus genotypes and correlation with viral load and risk factors in chronic positive patients.
Hepatitis C virus (HCV) has emerged as a leading cause of chronic hepatitis, liver cirrhosis and hepatocellular carcinoma worldwide. The purpose of this study was to describe the distribution pattern of HCV genotypes in chronic hepatitis patients in the Campania region of southern Italy and estimate their association with risk factors and viral load. 404 consecutive HCV ribonucleic acid-positive patients were included in the study. HCV genotyping was carried out by the HCV line probe assay test and viral load estimation by the TaqMan real-time PCR system. The predominant genotype was 1 (63.6%), followed by genotype 2 (29.4%), 3 (6.2%) and 4 (0.8%). Subtype 1b was more frequent in females than in males. Conversely, genotype 3 was more frequent in males. No significant difference was observed in age distribution of HCV genotypes. Surgery and dental therapy were the most frequent risk factors for genotype 1 and intravenous drug abuse and tattooing for genotype 3. Patients with genotype 1 more frequently showed high HCV viral load when compared to those with genotypes 2 and 3. The present study revealed that HCV genotypes 1 and 2 accounted for over 95% of all HCV infections in the Campania region, and genotype 1 was more frequently associated with a higher viral load when compared to genotypes 2 and 3.


Gene expression profiling of experimental saccular aneurysms using deoxyribonucleic acid microarrays.
The molecular characteristics of the pathophysiology of saccular aneurysms remain poorly understood. The purpose of the current study was to investigate the expression of various groups of genes at different stages of aneurysm age in elastase-induced saccular aneurysms in rabbits through the use of deoxyribonucleic acid (DNA) microarrays. A microarray consisting of genes related to cell adhesion, apoptosis, cell signaling, growth, inflammation, vascular remodeling, and oxidative stress was constructed by using rabbit nucleotide sequences. Elastase-induced saccular aneurysms were created at the origin of the right common carotid artery (CCA) in 12 rabbits. Two weeks (n=6) and 12 weeks (n=6) after aneurysm creation, ribonucleic acid (RNA) was isolated from the aneurysm and the control unoperated left CCA and was used for microarray experiments. Real-time polymerase chain reaction (RT-PCR) was performed for validation of microarray results. Of 209 genes, 157 (75%) at 2 weeks and 88 (42%) at 12 weeks demonstrated statistically significant differential expression between aneurysm tissue and the control left CCA tissue (P < .05). Multiple genes implicated in vessel wall remodeling were found to be elevated at 2 weeks and at 12 weeks. Expression of cell adhesion molecules and antioxidant enzymes was down-regulated at 2 weeks but was not significantly different from that of controls at 12 weeks. Most transcription factors, inflammatory genes, and structural genes showed underexpression at both time points. The expression profiles of selected genes were confirmed by RT-PCR. Multiple genes in diverse pathways have been differentially expressed in the rabbit aneurysm model.


Accurate prostate volume estimation using multifeature active shape models on T2-weighted MRI.
Accurate prostate volume estimation is useful for calculating prostate-specific antigen density and in evaluating posttreatment response. In the clinic, prostate volume estimation involves modeling the prostate as an ellipsoid or a spheroid from transrectal ultrasound, or T2-weighted magnetic resonance imaging (MRI). However, this requires some degree of manual intervention, and may not always yield accurate estimates. In this article, we present a multifeature active shape model (MFA) based segmentation scheme for estimating prostate volume from in vivo T2-weighted MRI. We aim to automatically determine the location of the prostate boundary on in vivo T2-weighted MRI, and subsequently determine the area of the prostate on each slice. The resulting planimetric areas are aggregated to yield the volume of the prostate for a given patient. Using a set of training images, the MFA learns the most discriminating statistical texture descriptors of the prostate boundary via a forward feature selection algorithm. After identification of the optimal image features, the MFA is deformed to accurately fit the prostate border. An expert radiologist segmented the prostate boundary on each slice and the planimetric aggregation of the enclosed areas yielded the ground truth prostate volume estimate. The volume estimation obtained via the MFA was then compared against volume estimations obtained via the ellipsoidal, Myschetzky, and prolated spheroids models. We evaluated our MFA volume estimation method on a total 45 T2-weighted in vivo MRI studies, corresponding to both 1.5 Tesla and 3.0 Tesla field strengths. The results revealed that the ellipsoidal, Myschetzky, and prolate spheroid models overestimated prostate volumes, with volume fractions of 1.14, 1.53, and 1.96, respectively. By comparison, the MFA yielded a mean volume fraction of 1.05, evaluated using a fivefold cross-validation scheme. A correlation with the ground truth volume estimations showed that the MFA had an r(2) value of 0.82, whereas the clinical volume estimation schemes had a maximum value of 0.70. Our MFA scheme involves minimal user intervention, is computationally efficient and results in volume estimations more accurate than state of the art clinical models.


Instant Detection of Synthetic Cannabinoids on Physical Matrices, Implemented on a Low-Cost, Ultraportable Device.
Synthetic cannabinoids (SCs) make up a class of novel psychoactive substances (NPS), used predominantly in prisons and homeless communities in the U.K. SCs can have severe side effects, including psychosis, stroke, and seizures, with numerous reported deaths associated with their use. The chemical diversity of SCs presents the major challenge to their detection since approaches relying on specific molecular recognition become outdated almost immediately. Ideally one would have a generic approach to detecting SCs in portable settings. The problem of SC detection is more challenging still because the majority of SCs enter the prison estate adsorbed onto physical matrices such as paper, fabric, or herb materials. That is, regardless of the detection modality used, the necessary extraction step reduces the effectiveness and ability to rapidly screen materials on-site. Herein, we demonstrate a truly instant generic test for SCs, tested against real-world drug seizures. The test is based on two advances. First, we identify a spectrally silent region in the emission spectrum of most physical matrices. Second, the finding that background signals (including from autofluorescence) can be accurately predicted is based on tracking the fraction of absorbed light from the irradiation source. Finally, we demonstrate that the intrinsic fluorescence of a large range of physical substrates can be leveraged to track the presence of other drugs of interest, including the most recent iterations of benzodiazepines and opioids. We demonstrate the implementation of our presumptive test in a portable, pocket-sized device that will find immediate utility in prisons and law enforcement agencies around the world.


Aberrant DNA methylation at genes associated with a stem cell-like phenotype in cholangiocarcinoma tumors.
Genetic abnormalities of cholangiocarcinoma have been widely studied; however, epigenomic changes related to cholangiocarcinogenesis have been less well characterized. We have profiled the DNA methylomes of 28 primary cholangiocarcinoma and six matched adjacent normal tissues using Infinium's HumanMethylation27 BeadChips with the aim of identifying gene sets aberrantly and epigenetically regulated in this tumor type. Using a linear model for microarray data, we identified 1610 differentially methylated autosomal CpG sites, with 809 hypermethylated (representing 603 genes) and 801 hypomethylated (representing 712 genes) in cholangiocarcinoma versus adjacent normal tissues (false-discovery rate ≤ 0.05). Gene ontology and gene set enrichment analyses identified gene sets significantly associated with hypermethylation at linked CpG sites in cholangiocarcinoma including homeobox genes and target genes of PRC2, EED, SUZ12, and histone H3 trimethylation at lysine 27. We confirmed frequent hypermethylation at the homeobox genes HOXA9 and HOXD9 by bisulfite pyrosequencing in a larger cohort of cholangiocarcinoma (n = 102). Our findings indicate a key role for hypermethylation of multiple CpG sites at genes associated with a stem cell-like phenotype as a common molecular aberration in cholangiocarcinoma. These data have implications for cholangiocarcinogenesis, as well as possible novel treatment options using histone methyltransferase inhibitors.


Dubious Heritage: Tobacco, History, and the Perils of Remembering the Rural Past .
In 1994 the Virginia legislature created a vehicle license plate to memorialize the state's long history of tobacco agriculture. Other states have likewise created plates to allow drivers to voice support for farmers. Like Virginia's "Tobacco Heritage" plate, many use traditional imagery and direct appeals to the history of agriculture. Given the United States' automobile culture, the plates are prominent, if small, works of public history that offer official sanction to a particular view of agriculture's past. In creating the license plate, the Virginia legislature valorized a particular telling of tobacco's past and is best understood as part of a much larger effort to employ history to deflect criticism of the tobacco industry. This effort, like the license plate, relied on simplified imagery that elided the complex history of tobacco in Virginia


ONJ in two dental practice-based research network regions.
The incidence of osteonecrosis of the jaw (ONJ) in the population is low, but specifics are unknown. Potential risk factors include bisphosphonate treatment, steroid treatment, osteoporosis, and head/neck radiation. This Dental Practice-Based Research Network study estimated ONJ incidence and odds ratios from bisphosphonate exposure and other risk factors using a key word search and manual chart reviews of electronic records for adults aged > or = 35 years enrolled during 1995-2006 in 2 large health care organizations. We found 16 ONJ cases among 572,606 cohort members; 7 additional cases were identified through dental plan resources. Among 23 cases (0.63 per 100,000 patient years), 20 (87%) had at least 1 risk factor, and 6 (26%) had received oral bisphosphonates. Patients with oral bisphosphonates were 15.5 (CI, 6.0-38.7) more likely to have ONJ than non-exposed patients; however, the sparse number of ONJ cases limits firm conclusions and suggests that the absolute risks for ONJ from oral bisphosphonates is low.


Assembly of the poorly differentiated Verasper variegatus W chromosome by different sequencing technologies.
The assembly of W and Y chromosomes poses significant challenges in vertebrate genome sequencing and assembly. Here, we successfully assembled the W chromosome of Verasper variegatus with a length of 20.48 Mb by combining population and PacBio HiFi sequencing data. It was identified as a young sex chromosome and showed signs of expansion in repetitive sequences. The major component of the expansion was Ty3/Gypsy. The ancestral Osteichthyes karyotype consists of 24 protochromosomes. The sex chromosomes in four Pleuronectiformes species derived from a pair of homologous protochromosomes resulting from a whole-genome duplication event in teleost fish, yet with different sex-determination systems. V. variegatus and Cynoglossus semilaevis adhere to the ZZ/ZW system, while Hippoglossus stenolepis and H. hippoglossus follow the XX/XY system. Interestingly, V. variegatus and H. hippoglossus derived from one protochromosome, while C. semilaevis and H. stenolepis derived from another protochromosome. Our study provides valuable insights into the evolution of sex chromosomes in flatfish and sheds light on the important role of whole-genome duplication in shaping the evolution of sex chromosomes.


Role of primary substrate composition and concentration on attenuation of trace organic chemicals in managed aquifer recharge systems.
This study was undertaken to investigate the role of primary substrate composition and concentration on the attenuation of biodegradable emerging trace organic chemicals (TOrCs) in simulated managed aquifer recharge (MAR) systems. Four sets of soil columns were established in the laboratory, each receiving synthetic feed solutions comprising different ratios and concentrations of peptone-yeast and humic acid as the primary substrate to investigate the effect on removal of six TOrCs (atenolol, caffeine, diclofenac, gemfibrozil, primidone, and trimethoprim). Based on abiotic control experiments, adsorption was not identified as a significant attenuation mechanism for primidone, gemfibrozil and diclofenac. Caffeine, atenolol and trimethoprim displayed initial adsorptive losses, however, adsorption coefficients derived from batch tests confirmed that adsorption was limited and in the long-term experiment, biodegradation was the dominant attenuation process. Within a travel time of 16 h, caffeine - an easily degradable compound exhibited removal exceeding 75% regardless of composition or concentration of the primary substrate. Primidone - a poorly degradable compound, showed no removal in any column regardless of the nature of the primary substrate. The composition and concentration of the primary substrate, however, had an effect on attenuation of moderately degradable TOrCs, such as atenolol, gemfibrozil and diclofenac, with the primary substrate composition seeming to have a larger impact on TOrC attenuation than its concentration. When the primary substrate consisted mainly of refractory substrate (humic acid), higher removal of the moderately degradable TOrCs was observed. The microbial communities in the columns receiving more refractory carbon, were noted to be more diverse and hence likely able to express a wider range of enzymes, which were more suitable for TOrC transformation. The effect of the primary substrate on microbial community composition, diversity and gene expression potential confirmed its influence on TOrC degradation.


Male breast cancer in Singapore: 15 years of experience at a single tertiary institution.
Male breast cancer is a rare disease entity, with little data from the Southeast Asian perspective. Hence, this study aims to review the data from our local experience in order to better delineate the disease characteristics in our population. Male patients with histologically proven breast cancer were identified from a prospectively collected database. The clinical, histopathological and survival data were reviewed retrospectively and analysed. Twenty-one patients were identified. The median age at diagnosis was 68 years. Eighteen patients underwent simple mastectomy with curative intent, with the remaining patients having metastatic disease at presentation. Almost half of the patients presented with stage III or IV disease. At the time of analysis, median overall survival was 50 months and median disease-free survival was 47.5 months. None of the patients had any documented family history or risk factors for male breast cancer. The disease appears to be a sporadic and rare occurrence in the local male population. A high index of suspicion should be maintained in males presented with a unilateral breast lump so that appropriate treatment can be instituted.


Frequency of nasal carriage of Staphylococcus aureus and its antimicrobial resistance pattern in patients on hemodialysis.
Staphylococcus aureus is currently the most common cause of infection in hospitalized patients. Patients on hemodialysis are at increased risk due to their immunocompromised state. The present study was designed to determine the frequency of S aureus nasal carriage in patients on hemodialysis. This study was undertaken in 2 dialysis centers to establish the frequency of S aureus nasal carriage at teaching hospitals of Mazandaran University of Medical Sciences, in the north of Iran. Standardized nose swabs were rotated into the anterior nares of the patients, and the samples were cultured on a blood-agar medium. Having grown the colony, gram stain, catalase, manitol, DNAase, and coagulase tests were all performed. Pattern of antibacterial sensitivity was determined by using the disc diffusion method. Also, agar dilution method was used to determine minimal inhibitory concentration of oxacillin and vancomycin. Of 84 patients on hemodialysis, 31 (36.9%) were nasal carriers of S aureus, of whom 23 (74.2%) were resistant to methicillin. Of the methicillin-resistant S aureus isolates, 3 (13.0%) were resistant to vancomycin and 7 (56.5%) had reduced susceptibility to vancomycin in agar dilution method. Resistance frequencies to clindamycin, ciprofloxacin, and trimethoprim-sulfamethoxsazole were 12.9%, 9.7%, and 19.3%, respectively. However, all microorganisms were sensitive to rifampicin. Patients on hemodialysis are at an increased risk of S aureus infections; thus, screening of these susceptible patients should be considered as a health priority. Meanwhile, microbial sensitivity tests should be ordered for all cases in order to optimize treatment options.


EZH2 inhibitors transcriptionally upregulate cytotoxic autophagy and cytoprotective unfolded protein response in human colorectal cancer cells.
Enhancer of zeste homolog 2 (EZH2) has been emerged as novel anticancer target. Various EZH2 small-molecule inhibitors have been developed in recent years. A major class of EZH2 inhibitors are S-adenosyl-L-methionine (SAM)-competitive inhibitors, such as EPZ005687, EI1, GSK126, UNC1999 and GSK343. Autophagy, a physiological process of self-digestion, is involved in the turnover of proteins or intracellular organelles. It can serve as cytoprotective or cytotoxic function in cancer. Our previous study has found that UNC1999 and GSK343 are potent autophagy inducers. In this study, the underlying molecular mechanisms were further investigated. Our results showed that UNC1999 and GSK343 transcriptionally upregulated autophagy of human colorectal cancer (CRC) cells through inducing LC3B gene expression. Besides, UNC1999/GSK343-induced autophagy was partially dependent on ATG7 but independent to EZH2 inhibition. Microarray and PCR array analyses identified that UNC1999 and GSK343 also induced endoplasmic reticulum (ER) stress and unfolded protein response (UPR). UNC1999/GSK343-induced ER stress/UPR contributed to the survival of cancer cells, which was opposite to UNC1999/GSK343-induced autophagy that promoted cell death.


Epidemiology of Noble Pen Shell (Pinna nobilis L. 1758) Mass Mortality Events in Adriatic Sea Is Characterised with Rapid Spreading and Acute Disease Progression.
From May to October 2019, multiple mass mortality events (MMEs) of Pinna nobilis were observed along Croatian coastline starting from the south-east and rapidly progressing in north-western direction. Time dynamics of the MMEs closely followed general speed and direction patterns of surface sea-currents, advancing approximately 350 km in less than 3 months. Surveillance, clinical evaluation, and sample collection were performed on multiple sites with various degrees of mortality rates. Moribund P. nobilis individuals were collected and subjected to pathological, molecular, and microscopical investigation. Affected animals were positive for Mycobacterium in 70% of the individuals, and Haplosporidium pinnae was present in 58% of the cases. Observed pathological lesions were most severe where concurrent presence of both pathogens was confirmed (in 45.8% of moribund individuals). Moderate to strong lesions were observed in animals positive for Mycobacterium only (25% of cases), and lesions were absent or minor to moderate when only H. pinnae was confirmed (16% of cases). Considering the rapid and severe spread of the MMEs, the areas less exposed to major sea currents appeared to be at lower risk of pathogen transmission. Surveillance activities along the Croatian coastline identified several P. nobilis populations in such "lower risk" areas without apparent mortality or clinical symptoms. Such areas are of particular interest as source of potentially healthy individuals to support active recovery actions.


The clinical effects of the venomous Lessepsian migrant fish Plotosus lineatus (Thunberg, 1787) in the Southeastern Mediterranean Sea.
Plotosus lineatus is a venomous fish that has migrated from the Indo-Pacific region to the Mediterranean Sea (Lessepsian migrant). Its presence in the Mediterranean Sea was first recorded in 2002 and was observed in growing schools. Its spines contain toxins with lytic, hemolytic and edematous activities. To characterize the injuries caused by Plotosus lineatus in the Southeastern Mediterranean Sea. A prospective observational case series of consultations provided by a national Poison Center pertaining to Plotosus lineatus from 2007 to 2016. Demographic and clinical data and method of fish identification were retrieved from the medical toxicological records, and described. Eighty four cases were included; the main findings are: median age 35 (range 3-80) years, 91.7% males, 51.2% fishermen, 78.6% palm injuries, 94% and 4.8% were mildly and moderately injured, respectively. Main local manifestations included pain, puncture wound, swelling, and erythema (90.5%, 70.2%, 33.3%, and 16.7%, respectively). Systemic signs were minor and infrequent (≤7.1%), including hypertension, tachycardia, vomiting, chills, and weakness. Management included wound disinfection, immersion in hot water, tetanus prophylaxis, and analgesics. No patient required hospital admission. The fish was identified mostly by the victim with the aid of the Poison Center (mainly by typical description, and a picture), and some by marine biologists. Plotosus lineatus is a new fish in the Southeastern Mediterranean Sea. It affects fishermen handling fishing nets, and beach hikers stepping on or holding it. Injuries caused by its spines usually result in minor effects; pain may be intense. Treatment includes disinfection, analgesics, and antitetanus and antibiotics as needed. No lethal cases were recorded, unlike exposure of animals to the venom of the Indo-Pacific species; reason is unclear. Our series illustrates the consequences of manmade disruption of ecosystem resulting in invasion of toxic species to a new environment, affecting human health.


Antemortem diagnosis and successful management of noncompressive segmental myelopathy in a Siberian-Bengal mixed breed tiger.
A 10-yr-old female spayed mixed breed tiger presented for a 9-day history of acute and nonprogressive paralysis of the pelvic limbs. Magnetic resonance imaging revealed a lesion suggestive of fibrocartilaginous embolic myelopathy with regional spinal cord edema, decreased disk signal intensity at L2-L3, and mild intervertebral disk protrusion at L1-L2 and L2-L3. Cerebral spinal fluid analysis showed no overt evidence of infection or neoplasia. Medical therapy was instituted, including corticosteroids and gastroprotectants as well as nursing care and physical therapy. The tiger began showing clinical improvement 2 wk after initiating treatment, progressing to the point where the animal was standing and intermittently walking. Three months after diagnosis, the tiger had regained muscle strength of its hind limbs and walked regularly with improving coordination. This case is the first report of antemortem diagnosis and successful medical management of suspected fibrocartilaginous embolic myelopathy in a large exotic felid.


[Drug prices: how they are established and existing price control systems].
Price is one of the main barriers of access to medicines. It is therefore important to understand how prices are formed and what factors determine the amount, as well as what interventions and regulations are the most appropriate considering their effects on access, innovation, local production and other potential objectives of drug policy. Economic analysis has developed a set of market models that can explain the behavior of prices, although actual markets diverge substantially from the theoretical models. Price regulation is justified by the so-called "market failures." Price regulation based on the cost of production, the most traditional form of price control, has fallen into disuse in favor of systems of international reference pricing and value-based pricing.


Modifying behaviour to reduce over-speeding in work-related drivers: an objective approach.
The goal of this study was to utilise an objective measurement tool, via an on-board Diagnostic tool (OBDII), to explore the effectiveness of a behaviour modification intervention designed to reduce over-speed violations in a group of work-related drivers. It was predicted that over-speed violations would be decreased following participation in a behaviour modification intervention where drivers received weekly feedback on their speeding performance and goal setting exercises. The final analysis included the on-road behaviour of 16 drivers, all of whom completed each stage of the intervention programme. As predicted, over-speed violations significantly decreased from pre-test to post-test, after controlling for kilometres driven. These findings offer practical guidance for industry in developing interventions designed to improve work-related driving behaviour.


Treatment of intestinal pythiosis in a dog with a combination of marginal excision, chemotherapy, and immunotherapy.
A 1.5-year-old mixed-breed dog was examined because of a 1-month history of anorexia, vomiting, diarrhea, and weight loss. The dog was very thin on physical examination (body condition score, 3/9). Results of all diagnostic tests were within reference limits except intestinal thickening and lymphadenopathy were identified on abdominal ultrasound examination. During exploratory laparotomy, thickening at the ileocecal-colic junction and within the transverse colon and mesenteric lymphadenopathy were identified, and the ileocecal-colic junction was resected. Histopathologic evaluation of the ileocecal-colic junction and full-thickness biopsy specimens from other sites as well as results of a serum ELISA were diagnostic for gastrointestinal Pythium insidiosum infection. Pythiosis was initially treated medically with administration of itraconazole and terbinafine by mouth, but the colonic lesion was progressive with this regimen. Two months after diagnosis, a subtotal colectomy was performed; marginal excision (0.6 cm) was obtained at the aboral margin. The dog was treated with 3 doses of a pythiosis vaccine beginning approximately 2 weeks after surgery and was continued on itraconazole and terbinafine for 5 months. Parenteral and enteral nutrition as well as considerable general supportive care were required postoperatively. Six months after treatment, the dog had a normal serum ELISA titer. Two years after treatment, the dog had returned to preoperative weight and was clinically normal. This patient had an unusually positive therapeutic response to chronic, extensive, marginally excised gastrointestinal pythiosis.


Risk factors of heart failure among patients with hypertension attending a tertiary hospital in Ibadan, Nigeria: The RISK-HHF case-control study.
Hypertension is the leading cause of heart failure (HF) in sub-Saharan Africa. Preventive public health approach to reduce the scourge of HF must seek to understand the risk factors of HF in at-risk populations. The aim of this study was to characterize the risk factors of HF among patients with hypertension attending a cardiology clinic. One hundred and one (101) case-control age- and sex-matched pairs were recruited. The study population were adults with a clinical diagnosis of hypertensive HF (cases) and individuals with systemic hypertension without HF. They were interviewed and evaluated for cardiovascular risk factors. Associations between variables were tested with chi square test, Fisher's exact test and independent sample t test as appropriate. Logistic regression modelling was used to determine the independent risk factors of hypertensive HF (HHF) in the study population while 'punafcc' package in stata12 was used to calculate the population attributable fraction (PAF) of the risk factors. Suboptimal medication adherence was the strongest adverse risk factor of HHF (medium adherence aOR: 3.53, 95%CI: 1.35-9.25; low adherence aOR: 9.44, 95%CI: 3.41-26.10) with a PAF of 67% followed by dipstick proteinuria (aOR: 4.22, 95%CI: 1.62-11.02; PAF: 34%) and alcohol consumption/day per 10grams (aOR: 1.23, 95%CI: 1.02-1.49; PAF: 22%). The protective risk factors of HHF were use of calcium channel blockers (aOR 0.25, 95%CI: 0.11-0.59; PAF: 59%), then daily fruits and vegetable consumption (aOR 0.41, 95%CI: 0.17-1.01; PAF: 46%), and eGFR (aOR 0.98, 95%CI: 0.96-0.99; PAF: 5.3%). The risk factors of HHF are amenable to lifestyle and dietary changes. Public health interventions and preventive cardiovascular care to improve medication adherence, promote fruit and vegetable consumption and reduce alcohol consumption among patients with hypertension are recommended. Renoprotection has utility in the prevention of HF among hypertensives.


Movement Component Analysis of Reaching Strategies in Individuals With Stroke: Preliminary Study.
Upper limb motor paresis is a major symptom of stroke, which limits activities of daily living and compromises the quality of life. Kinematic analysis offers an in-depth and objective means to evaluate poststroke upper limb paresis, with anticipation for its effective application in clinical settings. This study aims to compare the movement strategies of patients with hemiparesis due to stroke and healthy individuals in forward reach and hand-to-mouth reach, using a simple methodology designed to quantify the contribution of various movement components to the reaching action. A 3D motion analysis was conducted, using a simplified marker set (placed at the mandible, the seventh cervical vertebra, acromion, lateral epicondyle of the humerus, metacarpophalangeal [MP] joint of the index finger, and greater trochanter of the femur). For the forward reach task, we measured the distance the index finger's MP joint traveled from its starting position to the forward target location on the anterior-posterior axis. For the hand-to-mouth reach task, the shortening of the vertical distance between the index finger MP joint and the position of the chin at the start of the measurement was measured. For both measurements, the contributions of relevant upper limb and trunk movements were calculated. A total of 20 healthy individuals and 10 patients with stroke participated in this study. In the forward reach task, the contribution of shoulder or elbow flexion was significantly smaller in participants with stroke than in healthy participants (mean 52.5%, SD 24.5% vs mean 85.2%, SD 4.5%; P<.001), whereas the contribution of trunk flexion was significantly larger in stroke participants than in healthy participants (mean 34.0%, SD 28.5% vs mean 3.0%, SD 2.8%; P<.001). In the hand-to-mouth reach task, the contribution of shoulder or elbow flexion was significantly smaller in participants with stroke than in healthy participants (mean 71.8%, SD 23.7% vs mean 90.7%, SD 11.8%; P=.009), whereas shoulder girdle elevation and shoulder abduction were significantly larger in participants with stroke than in healthy participants (mean 10.5%, SD 5.7% vs mean 6.5%, SD 3.0%; P=.02 and mean 16.5%, SD 18.7% vs mean 3.0%, SD 10.4%; P=.02, respectively). Compared with healthy participants, participants with stroke achieved a significantly greater distance via trunk flexion in the forward reach task and shoulder abduction and shoulder girdle elevation in the hand-to-mouth reach task, both of these differences are regarded as compensatory movements. Understanding the characteristics of individual motor strategies, such as dependence on compensatory movements, may contribute to tailored goal setting in stroke rehabilitation.


Field Research and Parametric Analysis in a Medical-Surgical Unit.
To study the workplace in a medical-surgical (med-surg) unit and to identify suboptimal environmental conditions that can be improved in the current unit and avoided in future design, through rapidly deployed field research and timely simulation. Literature emphasizes the importance of the healthcare workplace and the effect on patient outcomes. What is lacking are studies conducted on-site and used for immediate application in design to assess and improve workplace conditions. A rapidly deployed field research and simulation study was conducted in a 40-bed med-surg unit of a large healthcare system as part of the process of designing a new medical tower. Online surveys, systematic behavioral observations, semi-structured interviews, sound studies, and advanced spatial analysis through parametric modeling were conducted. The following created challenges for patient monitoring, care coordination, and management: (1) waste and variability in walking, (2) limited point-of-use access to supplies, (3) large distances traveled for minor tasks, and (4) low visibility and connectivity. The corridor is used as a workspace/communication hub. There is a distinct difference in beginning of day and night shift patterns and between walking "distance" and walking "sequence." There is a tendency for nurses to multitask, but a simulation exercise shows that for key tasks like medication delivery, multitasking may not always reduce walking distances. Co-location of medications, supplies, and nourishment; accommodation for work on wheels; and spatial and technological connectivity between care team and patients should be considered while designing a med-surg unit. Understanding the key activity sequences helps determine the proximity of spaces in relationship to patient rooms and each other.


Once-weekly D-cycloserine effects on negative symptoms and cognition in schizophrenia: an exploratory study.
Daily dosing with d-cycloserine has inconsistently improved negative symptoms in schizophrenia patients, whereas intermittent dosing significantly facilitated exposure-based therapy in two studies of patients with phobic anxiety. In animal models, single-dose administration enhances memory consolidation, but tachyphylaxis develops with repeated dosing. The objective of this exploratory study was to assess whether once-weekly dosing with d-cycloserine will produce persistent improvements in negative symptoms and cognition. Fifty stable adult schizophrenia outpatients treated with any antipsychotic except clozapine were enrolled and 38 were randomized, double-blind, in a parallel-group, eight-week add-on trial of d-cycloserine 50 mg or placebo administered once-weekly. Symptom rating scales and a cognitive battery were administered at baseline and week 8 before the dose of study drug. As an exploratory analysis of memory consolidation, the Logical Memory Test, modified to measure recall after 7 days, was administered at baseline and after the first weekly dose of d-cycloserine. The primary outcome measures were change from baseline to week 8 on the SANS total score and on a composite cognitive score. Thirty-three subjects (87%) completed the trial. d-cycloserine significantly improved SANS total scores compared to placebo at week 8. Cognitive performance did not improve with d-cycloserine at 8 weeks. Delayed thematic recall on the Logical Memory Test was significantly improved with the first dose of d-cycloserine compared to placebo. Performance on immediate thematic recall and item recall on the Logical Memory Test did not differ between treatments. Once-weekly dosing with d-cycloserine for 8 weeks produced persistent improvement of negative symptoms compared to placebo, although statistical significance was, in part, the result of worsening of negative symptoms with placebo. Consistent with animal models, a single dose of d-cycloserine facilitated memory consolidation tested after 7 days on a test of thematic recall. These results must be considered preliminary since a number of outcomes were examined without correction for multiple tests. These findings suggest that once-weekly dosing with d-cycloserine for the treatment of negative symptoms merits further study, as do d-cycloserine effects on memory consolidation.


Agonist of growth hormone-releasing hormone as a potential effector for survival and proliferation of pancreatic islets.
Therapeutic strategies for transplantation of pancreatic islet cells are urgently needed to expand beta-cell mass by stimulating islet cell proliferation and/or prolonging islet cell survival. Control of the islets by different growth factors provides a potential venue for augmenting beta-cell mass. In the present study, we show the expression of the biologically active splice variant-1 (SV-1) of growth hormone-releasing hormone (GHRH) receptor in rat insulinoma (INS-1) cells as well as in rat and human pancreatic islets. In studies in vitro of INS-1 cells, the GHRH agonist JI-36 caused a significant increase in cell proliferation and a reduction of cell apoptosis. JI-36 increased islet size and glucose-stimulated insulin secretion in isolated rat islets after 48-72 h. At the ultrastructural level, INS-1 cells treated with agonist JI-36 revealed a metabolic active stimulation state with increased cytoplasm. Coincubation with the GHRH antagonist MIA-602 reversed the actions of the agonist JI-36, indicating the specificity of this agonist. In vivo, the function of pancreatic islets was assessed by transplantation of rat islets under the kidney capsule of streptozotocin-induced diabetic non-obese diabetic-severe combined immunodeficiency (NOD-SCID) mice. Islets treated with GHRH agonist JI-36 were able to achieve normoglycemia earlier and more consistently than untreated islets. Furthermore, in contrast to diabetic animals transplanted with untreated islets, insulin response to an i.p. glucose tolerance test (IPGTT) in animals receiving islets treated with agonist Jl-36 was comparable to that of normal healthy mice. In conclusion, our study provides evidence that agonists of GHRH represent a promising pharmacological therapy aimed at promoting islet graft growth and proliferation in diabetic patients.


Development and regulation of the phosphorylase-glycogen complex in myogenic cells of the L6 line.
The development of the phosphorylase-glycogen complex was studied in cells of the myogenic line L6 at different stages of differentiation. Our results indicate that the complex is already present in mononucleated myoblasts. However, several features of the complex observed in the myoblasts, such as the total amount of phosphorylase bound to the polysaccharide, and the type of glycogen agglomerates present in the cells, differ from those of the myotubes. It is postulated that at the moment when the myoblasts differentiate, definitive metabolic events take place. At this time the protein-glycogen complex is completed, phosphorylase being bound to small glycogen particles, and the multinucleated myotubes begin to accumulate a reserve of free glycogen. This latter constituent is mobilized first upon glucose starvation.


Structure and physical properties of new iron hydrogenophosphates: K2Fe(HP2O7)(H2PO4)2, LiH3Fe2(P2O7)2, and FeH2P2O7.
The exploration of the systems Fe-H-P-O, Li-Fe-H-P-O, and K-Fe-H-P-O using soft chemistry methods has allowed three new hydrogenophosphates to be synthesized, whose structures have been determined by ab initio calculations. The structures of two of them--FeH(2)P(2)O(7) and LiH(3)Fe(P(2)O(7))(2)--exhibit close relationships: their 3D framework consists of [FeO(4)](infinity) and [LiFe(2)O(12)](infinity) chains of edge-sharing octahedra, respectively, interconnected through diphosphate groups. These two diphosphates exhibit a paramagnetic to antiferromagnetic transition at low temperatures. The former phase exhibits intrachain ferromagnetic interactions (theta(p) > 0) in competition with the antiferromagnetic interchain ordering, whereas for the second one, the ferromagnetic interactions have disappeared due to the presence of Li in the chains. Differently, the third phosphate, K(2)Fe(HP(2)O(7))(H(2)PO(4))(2), exhibits a chain structure, involving isolated FeO(6) octahedra, and consequently is paramagnetic in the whole temperature range (4-300 K). Among these three phosphates, only the latter exhibits ionic conductivity, which may originate from the proton mobility.


Upcycling a plastic cup: one-pot synthesis of lactate containing metal organic frameworks from polylactic acid.
Waste PLA can be upcycled to metal organic frameworks of potential high value in a one-pot synthesis scheme, where PLA depolymerisation occurs in situ. Three homochiral lactate based frameworks were successfully synthesised and characterised from PLA as a feed source, including ZnBLD. The chiral separation ability of ZnBLD was maintained.


Fos-like immunoreactivity in the medulla after acute and chronic angiotensin II infusion.
Acute and chronic angiotensin (Ang) II hypertension are reported to have different mechanisms that involve differential contributions of the peripheral vasculature and the nervous system. Acute Ang II hypertension is mediated primarily by Ang acting at vascular smooth muscle, whereas chronic Ang II hypertension appears to have a neural component. In our experiments, the transition from a peripheral to a neural effect occurs over 10 hr of Ang II infusion in rats. To identify the role of the central nervous system in this transition, we measured Fos immunoreactivity, an indicator of neural activity, in the nucleus of the solitary tract (NTS), caudal ventrolateral medulla (CVL) and rostral ventrolateral medulla (RVL) in normal, sinoaortic denervated (SAD) and sham SAD rats after 2- or 18-hr Ang II infusion (50 ng/kg/min intravenously). Vehicle (5% dextrose) was infused in normal rats as control. Comparable increases in arterial pressure were produced by 2- and 18-hr Ang II infusion in all groups. Fos was increased in the NTS in sham SAD rats by 2- and 18-hr Ang II infusion (P < .05 vs. vehicle control). In the CVL, only 2-hr Ang II infusion was associated with increased Fos in normal and sham SAD rats (P < .05 vs. vehicle control) but not in SAD rats. In the RVL, 18-hr Ang II infusion elevated Fos in all groups (P < .05 vs. vehicle control). Activation of NTS during Ang II infusion is baroreceptor mediated and independent of infusion duration. Acute Ang II infusion produced a baroreceptor-mediated activation of the CVL, a region associated with baroreflex sympathoinhibition. Chronic Ang II infusion produced a baroreceptor-independent activation of the RVL, a brain area associated with sympathoexcitation, suggesting a centrally mediated increase in sympathetic outflow that may be associated with chronically infused Ang II.


Ageing in Escherichia coli requires damage by an extrinsic agent.
Evidence for ageing in symmetrically dividing bacteria such as Escherichia coli has historically been conflicting. Early work found weak or no evidence. More recent studies found convincing evidence, but negative results are still encountered. Because bacterial ageing is believed to result from non-genetic (e.g. oxidative) damage, we tested the possibility that the negative outcomes resulted from the lack of an extrinsic damage agent. We found that streptomycin, which produces mistranslated proteins that are more vulnerable to oxidation, was able to induce both damage and ageing in bacterial populations. A dosage effect relating the level of damage to the concentration of streptomycin was observed. Our results explain the previous inconsistencies, because all studies that failed to find evidence for bacterial ageing did not use a damage agent. However, all studies that succeeded in finding evidence utilized fluorescent proteins as a visual marker. We suggest that ageing in those studies was induced by the harmful effects of an extrinsic factor, such as the proteins themselves or the excitation light. Thus, all of the earlier studies can be reconciled and bacterial ageing is a real phenomenon. However, the study and observation of bacterial ageing require the addition of an extrinsic damage agent.


Constitutive Notch signalling promotes CD4 CD8 thymocyte differentiation in the absence of the pre-TCR complex, by mimicking pre-TCR signals.
Notch1 signalling is essential for the commitment of multipotent lymphocyte precursors towards the alphabeta T-cell lineage and plays an important role in regulating beta-selection in CD4(-)CD8(-) double-negative (DN) thymocytes. However, the role played by Notch in promoting the development of CD4(+)CD8(+) double-positive (DP) thymocytes is poorly characterized. Here, we demonstrate that the introduction of a constitutively active Notch1 (ICN1) construct into RAG(-/-) lymphocyte precursors resulted in the generation of DP thymocytes in in vitro T-cell culture systems. Notably, developmental rescue was dependent not only on the presence of an intact Notch1 RAM domain but also on Delta-like signals, as ICN1-induced DP development in RAG(-/-) thymocytes occurred within an intact thymus or in OP9-DL1 co-cultures, but not in OP9-control co-cultures. Interestingly, ICN1 expression in SLP-76(-/-) precursors resulted in only a minimal developmental rescue to the immature CD8(+) single-positive stage, suggesting that Notch is utilizing the same signalling pathway as the pre-TCR complex. In support of this, ICN1 introduction resulted in the activation of the ERK-MAPK-signalling cascade in RAG(-/-) thymocytes. Taken together, these studies demonstrate that constitutive Notch signalling can bypass beta-selection during early T-cell development by inducing pre-TCR-like signals within a T-cell-promoting environment.


Responses to sounds in the central auditory system of the frog: an advanced electrophysiology laboratory in sensory processing.
Frogs rely upon vocal communication to advertise for potential mates, to defend territory and to alarm neighbors of danger. Cells in the auditory midbrain of an awake frog display tuning to the spectral energy present in calls based upon discharge rate and encode the temporal properties of calls in the timing of their discharges. This laboratory experiment is designed to allow students to explore the relationship between stimulus amplitude or frequency and response rate, and how the timing of responses can also be used to encode behaviorally relevant features of the stimulus. Action potentials in the midbrain auditory nucleus, the torus semicularis, are evoked by delivery of free field sounds and recorded. Most cells are broadly tuned to frequency, yet some can be fairly precise in preserving periodic structure. The use of a comparative model of study should help students understand principles common among all sensory systems, and an appreciation that the architecture of each system is adaptively matched to the ethological task at hand.