Cellular Mechanisms of Singlet Oxygen in Photodynamic Therapy.
In this review, we delve into the realm of photodynamic therapy (PDT), an established method for combating cancer. The foundation of PDT lies in the activation of a photosensitizing agent using specific wavelengths of light, resulting in the generation of reactive oxygen species (ROS), notably singlet oxygen (1O2). We explore PDT's intricacies, emphasizing its precise targeting of cancer cells while sparing healthy tissue. We examine the pivotal role of singlet oxygen in initiating apoptosis and other cell death pathways, highlighting its potential for minimally invasive cancer treatment. Additionally, we delve into the complex interplay of cellular components, including catalase and NOX1, in defending cancer cells against PDT-induced oxidative and nitrative stress. We unveil an intriguing auto-amplifying mechanism involving secondary singlet oxygen production and catalase inactivation, offering promising avenues for enhancing PDT's effectiveness. In conclusion, our review unravels PDT's inner workings and underscores the importance of selective illumination and photosensitizer properties for achieving precision in cancer therapy. The exploration of cellular responses and interactions reveals opportunities for refining and optimizing PDT, which holds significant potential in the ongoing fight against cancer.


Mathematic modeling of human amniotic fluid dynamics.
We sought to develop a model quantifying the relative contributions of fetal swallowing and intramembranous flow to amniotic fluid dynamics during human gestation. We then used the model to simulate the impact of absent swallowing on amniotic fluid volume. The model was developed with published data for normal human amniotic fluid volume and composition, human fetal urine flow rate and composition (11 to 42 weeks), and extrapolated data from ovine lung fluid production. Fetal swallowing and intramembranous flow were calculated with assumptions that (1) swallowed fluid is isotonic to amniotic fluid, (2) intramembranous flow is free water diffusion, and (3) 50% of lung fluid is swallowed. The model was then applied to simulate absent fetal swallowing and variable (0%, 50%) proportions of swallowed lung fluid were used as a representation of esophageal atresia-tracheal fistula variations. Fetal swallowed volume and intramembranous flow linearly increase until 28 to 30 weeks. Daily swallowed volume then exponentially increases to a maximum of 1006 ml/day at term, whereas intramembranous flow continues on a linear trend to reach 393 ml/day at term. With absent swallowing and variable amounts of lung fluid swallowed (0%, 50%), predicted amniotic fluid volume is similar to normal values through 20 weeks, exceeds the 95% confidence interval for normal amniotic fluid volume at 29 to 30 weeks' gestation (approximately 2000 ml), and then exponentially increases. Predicted amniotic fluid osmolality (280 to 257 mOsm/kg) is slightly lower than actual values although within the clinically normal range. This model indicates that the normal reduction in amniotic fluid volume beginning at 34 weeks results from the marked increase in swallowed volume during the third trimester. Additionally, this model correlates well with the timing of the initial clinical presentation of polyhydramnios observed in some fetuses with conditions that result in absent or reduced swallowing or gastrointestinal atresia. Modeling of amniotic fluid dynamics can predict normal changes in fetal fluid exchange and may aid in understanding of amniotic fluid imbalances.


Experimental infection of chamois (Rupicapra pyrenaica parva) with Sarcoptes scabiei derived from naturally infected goats.
Two chamois (Rupicapra pyrenaica parva) out of a group of three were experimentally infected with Sarcoptes scabiei derived from a naturally infected domestic goat. One of the chamois presented the first clinical manifestations (papules and desquamation) at 7 days post-infection, after 22 days crusts and alopecia appeared and after 41 days pruritus. The other chamois presented desquamation after 15 days and papules after 21 days and crusts and alopecia after 31 days. All these clinical manifestations continued to spread and when the animals were treated at 84 days post-infection, pruritus, papules and crusts were first to disappear, there being no evidence of their presence at 99 days post-infection, when the second treatment dose was applied. The desquamation and alopecia disappeared at 114 days post-infection, by which stage both animals were considered to have been cured. The results of the skin scraping was negative in both chamois until 54 days post-infection and it became negative again after 84 days, when the first treatment dose was applied. Biopsies showed different levels of hyperkeratosis and a marked epidermic hyperplasia with formation of small crusts. Superficial epidermis presented marked vasodilatation and also infiltrated inflammation. None of the biopsies carried out showed the presence of parasites. The non-infected chamois, which was kept in the same compound as the other two, did not present any clinical manifestations compatible with infection by S. scabiei throughout the entire period of the experiment.


Volume-activated K+ and Cl- pathways of dissociated epithelial cells (MDCK): role of Ca2+.
Osmotic swelling of dissociated Madin-Darby canine kidney (MDCK) cells in NaCl medium is followed by shrinking (regulatory volume decrease, or RVD) or in KCl medium by secondary swelling. The cation ionophore gramicidin has little effect on volumes of isotonic cells but accelerates volume-activated changes in either medium. Immediately after hypotonic exposure, the membrane becomes transiently hyperpolarized followed by depolarization. The depolarization phase is diminished by the anion transport inhibitor 4,4'-diisothiocyanostilbene-2,2'-disulfonic acid (DIDS). Swelling is also associated with an almost immediate increase in Ca2+ influx and elevation of cytoplasmic Ca2+ ([Ca2+]i) preceding RVD. In Ca2(+)-free medium, [Ca2+]i rapidly declines to a low level. Osmotic swelling, under these circumstances, is associated with a small transient increase in [Ca2+]i, but RVD or secondary swelling (in KCl) are minimal. Under these conditions, addition of gramicidin or the Ca2(+)-ionophore A23187 induces significant volume changes, although not as large as those found in the presence of Ca2+. Quinine inhibits RVD in the absence of gramicidin, but not in its presence; oligomycin C, DIDS, and trifluoperazine, on the other hand, inhibit in the presence of the ionophore. These findings suggest that in MDCK cells RVD involves activation of distinct conductive K+ and Cl- pathways which allow escape of KCl and osmotically obligated water and that activation of both pathways is associated with elevated [Ca2+]i derived largely from volume activation of a Ca2(+)-influx pathway.


Cytolysins of Actinobacillus pleuropneumoniae serotype 9.
Cytolysin I (ClyI) and cytolysin II (ClyII), which are present in the culture supernatant of Actinobacillus pleuropneumoniae serotype 9, are thought to play an important role in the pathogenesis of pig pleuropneumonia. The purpose of this study was to clone and characterize the genetic determinants of these cytolysins. Cloning was accomplished by the screening of DNA libraries for the presence of cytolytic activity and for the presence of DNA sequences homologous to leukotoxin DNA of Pasteurella haemolytica. Both genetic determinants were found to be members of the RTX cytotoxin family. The ClyII determinant was characterized in more detail. It appeared that ClyII more closely resembled the leukotoxin of P. haemolytica than the alpha-hemolysin of Escherichia coli. The ClyII amino acid sequence was identical to a hemolysin gene sequence of A. pleuropneumoniae serotype 5; this finding indicates that the latter gene also codes for ClyII and not for ClyI, as has previously been suggested. The genetic organization of the ClyII determinant differed from the genetic organization of other RTX determinants. Genes responsible for secretion of ClyII were not contiguous with the toxin gene. Instead, secretion genes were present elsewhere in the genome. These secretion genes, however, belong to the ClyI operon. This indicates that the secretion genes of the ClyI operon are responsible for secretion of ClyI and ClyII.


[Illness and death: slaves in the city of Pelotas, 1870-1880].
The article analyzes diseases presented by slaves hospitalized at Santa Casa de Misericórdia in Pelotas. The focus is on the workers at 'charqueadas' (processing plants for dried meat), whose harsh and rigid work regimen had serious health consequences. Although we can find many descriptions of beef processing at 'charqueadas', we find less evidence of concerns about how slave labor was employed at these plants. By analyzing the period from 1870 to 1880, based on hospital records, travelers' observations, and newspaper reports, the article intends to contribute towards a better understanding of the health conditions of captives in the southern part of the state of Rio Grande do Sul.


Diversification of sympatric broadcast-spawning limpets (Cellana spp.) within the Hawaiian archipelago.
Speciation remains a central enigma in biology, and nowhere is this more apparent than in shallow tropical seas where biodiversity rivals that of tropical rainforests. Obvious barriers to gene flow are few and most marine species have a highly dispersive larval stage, which should greatly decrease opportunities for speciation via geographic isolation. The disparity in the level of geographic isolation for terrestrial and marine species is exemplified in Hawai'i where opportunities for allopatric speciation abound in the terrestrial realm. In contrast, marine colonizers of Hawai'i are believed to produce only a single endemic species or population, due to the lack of isolating barriers. To test the assertion that marine species do not diversify within Hawai'i, we examine the evolutionary origin of three endemic limpets (Cellana exarata, C. sandwicensis and C. talcosa) that are vertically segregated across a steep ecocline on rocky shores. Analyses of three mtDNA loci (12S, 16S, COI; 1565bp) and two nDNA loci (ATPSβ, H3; 709bp) in 26 Indo-Pacific Cellana species (N=414) indicates that Hawai'i was colonized once ∼3.4-7.2Ma from the vicinity of Japan. Trait mapping demonstrates that high-shore residence is the ancestral character state, such that mid- and low-shore species are the product of subsequent diversification. The Hawaiian Cellana are the first broadcast-spawners demonstrated to have speciated within any archipelago. The habitat stratification, extensive sympatry, and evolutionary history of these limpets collectively indicate a strong ecological component to speciation and support the growing body of evidence for non-allopatric speciation in the ocean.


Brain-type natriuretic peptide levels in the prediction of adverse outcome in patients with pulmonary embolism: a systematic review and meta-analysis.
The potential role of elevated brain-type natriuretic peptides (BNP) in the differentiation of patients suffering from acute pulmonary embolism at risk for adverse clinical outcome has not been fully established. We evaluated the relation between elevated BNP or N-terminal-pro-BNP (NT-pro-BNP) levels and clinical outcome in patients with pulmonary embolism. Articles reporting on studies that evaluated the risk of adverse outcome in patients with pulmonary embolism and elevated BNP or NT-pro-BNP levels were abstracted from Medline and EMBASE. Information on study design, patient and assay characteristics, and clinical outcome was extracted. Primary endpoints were overall mortality and predefined composite outcome of adverse clinical events. Data from 13 studies were included. In 51% (576/1,132) of the patients, BNP or NT-pro-BNP levels were increased. The different analyses were performed in subpopulations. Elevated levels of BNP or NT-pro-BNP were significantly associated with right ventricular dysfunction (P < 0.001). Patients with high BNP or NT-pro-BNP concentration were at higher risk of complicated in-hospital course (odds ratio [OR], 6.8; 95% confidence interval [CI], 4.4-10) and 30-day mortality (OR, 7.6; 95% CI, 3.4-17). Patients with a high NT-pro-BNP had a 10% risk of dying (68/671; 95% CI, 8.0-13%), whereas 23% (209/909; 95% CI, 20-26%) had an adverse clinical outcome. High concentrations of BNP distinguish patients with pulmonary embolism at higher risk of complicated in-hospital course and death from those with low BNP levels. Increased BNP or NT-pro-BNP concentrations alone, however, do not justify more invasive treatment regimens.


Wilms' tumours with intracaval involvement.
Since Gross established the basic rules for nephrectomy in Wilms' tumour (WT) in 1953, the management of nephroblastoma has been more straightforward. However, some cases with intravascular involvement, currently detected by ultrasound (US), may represent a daunting challenge for the surgeon. Inferior vena cava with tumour thrombus, induced by WT, can be asymptomatic and, if undetected, can contribute to poorer prognosis for two main reasons: possible neoplastic cells inside the thrombus and higher morbidity risk of surgery. From 1979 to 1993, 81 WT were studied by routine US. Intracaval thrombosis was diagnosed in four (5%), in one of which the thrombus extended to the right atrium. In our experience, the surgical strategy in each of the four cases (100% survival) depended on the length of the thrombus and whether or not it infiltrated the vena cava wall. If the thrombus can be easily removed: complete resection. However, in cases of atrial thrombus, and more particularly if the thrombus involves the intima, we suggest the thrombus not be touched since the problem may be solved by preoperative and postoperative chemotherapy. Thus the favourable prognosis would be maintained and superfluous risky surgery avoided.


Awareness and Acceptability of pre-exposure prophylaxis (PrEP) among gay, bisexual and other men who have sex with men (GBMSM) in Kenya.
Kenyan gay, bisexual, and other men who have sex with men (GBMSM) are significantly affected by HIV. Pre-exposure prophylaxis (PrEP) is an effective biomedical approach to HIV prevention. We conducted a cross-sectional survey of 459 HIV-negative Kenyan GBMSM to assess individual and interpersonal correlates of PrEP awareness/acceptability using univariate and hierarchical logistic regression modeling. We found that 64.3% of participants had heard of PrEP and 44.9% were willing to use PrEP. In hierarchical logistic regression models for PrEP awareness, condom use with regular partners, higher condom use self-efficacy, higher perceived ability to use PrEP, history of STI, and membership in LGBT organization were significantly associated with being aware of PrEP (χ2 = 69.6, p < .001). In hierarchical logistic regression models for PrEP acceptability, higher self-esteem, higher condom use self-efficacy, depression/anxiety, higher perceived ability to use PrEP, willingness to engage in PrEP follow-up visits, coercion at sexual debut, and family exclusion were significantly associated with being acceptable to PrEP (χ2 = 231.8, p < .001). Individual and interpersonal factors were significantly associated with PrEP awareness and acceptability. Our findings underscore the need to promote awareness and understanding of PrEP as an effective HIV prevention tool in combination with other safer-sex methods that are appropriate given an individual's personal circumstances.


Geographic Variation in the Initiation of Commonly Used Opioids and Dosage Strength in United States Nursing Homes.
To examine and quantify geographic variation in the initiation of commonly used opioids and prescribed dosage strength among older US nursing home residents. We merged 2011 Minimum Data Set 3.0 to Medicare claims and facility characteristics data to conduct a cross-sectional study among long-stay nursing home residents who initiated short-acting opioids commonly used in nursing homes (oxycodone, hydrocodone, or tramadol). We examined geographic variation in specific opioids initiated and potentially inappropriate doses (≥50 mg oral morphine equivalent/d) across hospital referral regions (HRRs). Multilevel logistic models quantified the proportional change in between-HRR variation and associations between commonly initiated opioids and inappropriate doses after adjusting for resident characteristics, facility characteristics, and state. Oxycodone (9.4%) was initiated less frequently than hydrocodone (56.2%) or tramadol (34.5%) but varied dramatically between HRRs (range, 0%-74.5%). In total, resident/facility characteristics and state of residence, respectively explained 84.1%, 58.2%, 59.1%, and 46.6% of the between-HRR variation for initiating oxycodone, hydrocodone, tramadol, and inappropriate doses. In all cases, state explained the largest proportion of between-HRR variation. Relative to hydrocodone, residents initiating oxycodone were more likely (adjusted odds ratio, 5.00; 95% confidence interval, 4.57-5.47) and those initiating tramadol were less likely (adjusted odds ratio, 0.28; 95% confidence interval, 0.25-0.31) to be prescribed potentially inappropriately high doses. We documented extensive geographic variation in the opioid and dose initiated for nursing home residents, with state explaining the largest proportion of the observed variation. Further work is needed to understand potential drivers of opioid prescribing patterns at the state level.


Myths and realities in cancer care: another point of view.
Cancer care is advancing, and the cancer community is right to celebrate that progress. Simultaneously, however, the cost of cancer therapy is rising along with all medical costs. In a matter of just a few years, the cost of health coverage is projected to reach heights that are simply unsustainable for most American families, overshadowing every other daily expense. Such an overwhelming burden will continue to undermine the progress made in developing new, more effective treatment and care for cancer patients. There cannot be a celebration of discoveries if there is no way to pay for them. It is from that perspective that I react and respond here to the commentary of Dana Goldman and Tomas Philipson, who debunk five myths in cancer care. I concur with the authors' assessment of the value and impact of progress in cancer care. However, I also argue that researchers and policy makers must pay more attention to the impact of cost on patients' adherence and experience and the speed with which new treatment and care are being developed.


The RB protein family in retinal development and retinoblastoma: new insights from new mouse models.
The Rb gene was isolated almost 20 years ago, but fundamental questions regarding its role in retinal development and retinoblastoma remain. What is the normal function of RB protein in retinogenesis? What is the cell-of-origin of retinoblastoma? Why do retinoblastoma tumors have recurrent genetic lesions other than Rb inactivation? Why is retinoblastoma not induced by defects in cell cycle regulators other than Rb? Why is the retina so sensitive to Rb loss? Recently developed conditional Rb knockout models provide new insight into some of these issues. The data suggest that RB protein may not control the rate of progenitor division, but is critical for cell cycle exit when dividing retinal progenitors differentiate into postmitotic transition cells. This finding focuses attention on the ectopically dividing transition cell, rather than the progenitor, as the cell-of-origin. Cell-specific analyses in the RB-deficient retina reveal that ectopically dividing photoreceptors, bipolar and ganglion cells die, but amacrine, horizontal and Muller cells survive and stop dividing when they terminally differentiate. Rare amacrine transition cells escape cell cycle exit and generate tumors. These data suggest that post-Rb mutations are required to overcome growth arrest associated with terminal differentiation, rather than apoptosis as previously suggested. To explain why perturbing cell cycle regulators other than RB does not initiate retinoblastoma, we speculate that mutations in other components of the RB pathway perturb cell cycle arrest, but only RB loss triggers genome instability in retinal transition cells, which may be critical to facilitate post-Rb mutations necessary for transformation. Cell-specific differences in the effect of Rb loss on genome stability may contribute to the tremendous sensitivity of retinal transition cells to tumorigenesis. The new mouse models of retinoblastoma will be invaluable for testing these possibilities.


Glial cell line-derived neurotrophic factor and somatostatin levels in cerebrospinal fluid of patients affected by chronic migraine and fibromyalgia.
The aim of the present study was to verify cerebrospinal fluid (CSF) levels of glial cell line-derived neurotrophic factor (GDNF) and somatostatin, both measured by sensitive immunoassay, in: 16 chronic migraine (CM) patients, 15 patients with an antecedent history of migraine without aura diagnosed as having probable chronic migraine (PCM) and probable analgesic-abuse headache (PAAH), 20 patients affected by primary fibromyalgia syndrome (PFMS), and 20 control subjects. Significantly lower levels of GDNF and somatostatin were found in the CSF of both CM and PCM + PAAH patients compared with controls (GDNF =P < 0.001, P < 0.002; somatostatin = P < 0.002, P < 0.0003), without significant difference between the two groups. PFMS patients, with and without analgesic abuse, also had significantly lower levels of both somatostatin and GDNF (P < 0.0002, P < 0.001), which did not differ from those of CM and PCM + PAAH patients. A significant positive correlation emerged between CSF values of GDNF and those of somatostatin in CM (r = 0.70, P < 0.02), PCM + PAAH (r = 0.78, P < 0.004), and PFMS patients (r = 0.68, P < 0.008). Based on experimental findings, it can be postulated that reduced CSF levels of GDNF and somatostatin in both CM and PCM + PAAH patients can contribute to sustained central sensitization underlying chronic head pain. The abuse of simple or combination analgesics does not seem to influence the biochemical changes investigated, which appear to be more strictly related to the chronic pain state, as demonstrated also for fibromyalgia.


Hypoxia modulates the differentiation potential of stem cells of the apical papilla.
Stem cells from the apical papilla (SCAP) are a population of mesenchymal stem cells likely involved in regenerative endodontic procedures and have potential use as therapeutic agents in other tissues. In these situations, SCAP are exposed to hypoxic conditions either within a root canal devoid of an adequate blood supply or in a scaffold material immediately after implantation. However, the effect of hypoxia on SCAP proliferation and differentiation is largely unknown. Therefore, the objective of this study was to evaluate the effect of hypoxia on the fate of SCAP. SCAP were cultured under normoxia (21% O2) or hypoxia (1% O2) in basal or differentiation media. Cellular proliferation, gene expression, differentiation, and protein secretion were analyzed by live imaging, quantitative reverse-transcriptase polymerase chain reaction, cellular staining, and enzyme-linked immunosorbent assay, respectively. Hypoxia had no effect on SCAP proliferation, but it evoked the up-regulation of genes specific for osteogenic differentiation (runt-related transcription factor 2, alkaline phosphatase, and transforming growth factor-β1), neuronal differentiation ( 2'-3'-cyclic nucleotide 3' phosphodiesterase, SNAIL, neuronspecific enolase, glial cell-derived neurotrophic factor and neurotrophin 3), and angiogenesis (vascular endothelial growth factor A and B). Hypoxia also increased the sustained production of VEGFa by SCAP. Moreover, hypoxia augmented the neuronal differentiation of SCAP in the presence of differentiation exogenous factors as detected by the up-regulation of NSE, VEGFB, and GDNF and the expression of neuronal markers (PanF and NeuN). This study shows that hypoxia induces spontaneous differentiation of SCAP into osteogenic and neurogenic lineages while maintaining the release of the proangiogenic factor VEGFa. This highlights the potential of SCAP to promote pulp-dentin regeneration. Moreover, SCAP may represent potential therapeutic agents for neurodegenerative conditions because of their robust differentiation potential.


Cleavage of transcriptional activator Oct-1 by poliovirus encoded protease 3Cpro.
In HeLa cells, RNA polymerase II mediated transcription is severely inhibited by poliovirus infection. Both basal and activated transcription are affected to bring about a complete shutoff of host cell transcription. We demonstrate here that the octamer binding transcription factor, Oct-1, is cleaved in HeLa cells infected with poliovirus. Incubation of Oct-1 with the purified, recombinant 3Cpro results in the generation of the cleaved Oct-1 product seen in virus infected cells. Poliovirus infection leads to the formation of altered Oct-1 DNA complexes that can also be generated by incubation of Oct-1 with purified 3Cpro. We also show that Oct-1 cleaved by 3Cpro loses its ability to inhibit transcriptional activation by the SV40 B enhancer. These results suggest that cleavage of Oct-1 in poliovirus infected cells leads to the loss of its activity.


Potential of Gentiana lutea for the Treatment of Obesity-associated Diseases.
Obesity, diabetes, and associated diseases are increasing all over the world, and pose a great burden on public health. According to the latest reports, 440 million people are suffering from diabetes. Diabetes is caused by impaired ability to produce or respond to the hormone insulin consequently resulting in hyperglycemia. Data used for this review was obtained by using PUBMED/MEDLINE (1987-2018). The main data search terms were: Gentiana lutea, Gentiana lutea extract, Gentiana lutea constituents, obesity, diabetes mellitus, diabetic complications. In the present review, we describe the potential of root powder of yellow gentian (Gentiana lutea) for the prevention of obesity and diabetes including complications related to this disease. Reasonably effective, low-cost alternatives could fulfill an important role for a large part of the human population and could be of great value for the food market. Even a modest reduction of morbidity and mortality with respect to this disease translates into millions of lives saved.


Prolonged CD4 depletion after sequential autologous peripheral blood progenitor cell infusions in children and young adults.
Administration of mobilized peripheral blood progenitor cells (PBPCs) after high-dose chemotherapy rapidly restores multilineage hematopoiesis, but the ability of such products to restore lymphocyte populations remains unclear. In this report, we evaluated immune reconstitution in a series of patients treated with sequential cycles of high-dose chemotherapy, followed by autologous PBPC infusions (median CD34(+) cell dose 7.2 x 10(6) cells/kg [range 2-29.3]). Although patients experienced rapid reconstitution of B cells and CD8(+) T cells, we observed CD4 depletion and diminished immune responsiveness in all patients for several months after completion of therapy. Mature CD4(+) T cells contained within the grafts did not appear to contribute substantially to immune reconstitution because CD4 counts did not differ between recipients of unmanipulated T-cell replete infusions versus CD34 selected, T-cell-depleted infusions. Rather, at 12 months after therapy, total CD4 count was inversely proportional to age (rho = -0.78, P =.04), but showed no relationship to CD34 cell dose (rho = -0.42, P =.26), suggesting that age-related changes within the host are largely responsible for the limited immune reconstitution observed. These results demonstrate that in the autologous setting, the infusion of large numbers of PBPCs is not sufficient to restore T-cell immune competence and emphasize that specific approaches to enhance immune reconstitution are necessary if immune-based therapy is to be used to eradicate minimal residual disease after autologous PBPC transplantation. (Blood. 2000;96:754-762)


New direct-acting antivirals for hepatitis C treatment and neuropsychiatric symptoms in psychiatric risk groups.
The new direct-acting antivirals (DAAs) have changed chronic hepatitis C virus (HCV) treatment perspectives by achieving success rates in all genotypes and by reducing the associated adverse effects in comparison to pegylated interferon α regimens. These adverse effects include depression, insomnia and suicidal intention, which make it difficult to treat psychiatric risk groups. The objectives of this study were to evaluate the neuropsychiatric symptoms during HCV treatment with DAAs in standard multidisciplinary clinical practice in psychiatric risk groups and to assess the risk factors associated with neuropsychiatric symptoms in this study population. A prospective cross-sectional study was performed of all consecutive patients with psychiatric risk and HCV infection who completed treatment with the new DAAs at the ambulatory care pharmacy. We recorded demographic and clinical data, neuropsychiatric symptoms, emergency consultations, admissions to the psychiatric unit and interventions to manage neuropsychiatric symptoms. Statistical analysis was used to assess the association between neuropsychiatric symptoms and clinical data. We included 48 patients with psychiatric risk and detected 36 neuropsychiatric symptoms in 17 (35.4%) patients, with a mean of 0.75 neuropsychiatric symptoms per patient. However, no studied risk factors for developing neuropsychiatric symptoms were found in this population. We found a moderate prevalence of neuropsychiatric symptoms in the psychiatric risk group. The neuropsychiatric symptoms more frequently reported were insomnia (17%), irritability (15%) and depression (13%). Neuropsychiatric symptoms in patients with psychiatric risk can occur during treatment with DAAs but less frequently than with pegylated interferon α regimens, and so is a safe treatment for these patients.


Female and male obese Zucker rats display differential inflammatory mediator and long non-coding RNA profiles.
The goal of this study was to identify mediators in peri-lymphatic adipose tissue (PLAT) that are altered in obese versus lean Zucker rats, with focus on potential sex differences MAIN METHODS: Mesenteric PLAT was analyzed with protein and lncRNA arrays. Additional RT-PCR confirmation was performed with epididymal/ovarian fat. MCP-1, TCK-1, Galectin-1, Galectin-3, and neuropilin-1 were elevated in PLAT from obese rats of both sexes. However, 11 additional proteins were elevated only in obese males while 24 different proteins were elevated in obese females. Profiling of lncRNAs revealed lean males have elevated levels of NEAT1, MALAT1 and GAS5 compared to lean females. NEAT1, MALAT1, and GAS5 were significantly reduced with obesity in males but not in females. Another lncRNA, HOTAIR, was higher in lean females compared to males, and its levels in females were reduced with obesity. Obese rats of both sexes had similar histologic findings of mesenteric macrophage crown-like structures and hepatocyte fat accumulation. While obese male and female Zucker rats both have increased inflammation, they have distinct signals. Future studies of the proteome and lncRNA landscape of obese males vs. females in various animal models and in human subjects are warranted to better guide development of therapeutics for obesity-induced inflammation.


Impaired Endothelium-Dependent Hyperpolarization Underlies Endothelial Dysfunction during Early Metabolic Challenge: Increased ROS Generation and Possible Interference with NO Function.
Endothelial dysfunction is a hallmark of diabetic vasculopathies. Although hyperglycemia is believed to be the culprit causing endothelial damage, the mechanism underlying early endothelial insult in prediabetes remains obscure. We used a nonobese high-calorie (HC)-fed rat model with hyperinsulinemia, hypercholesterolemia, and delayed development of hyperglycemia to unravel this mechanism. Compared with aortic rings from control rats, HC-fed rat rings displayed attenuated acetylcholine-mediated relaxation. While sensitive to nitric oxide synthase (NOS) inhibition, aortic relaxation in HC-rat tissues was not affected by blocking the inward-rectifier potassium (Kir) channels using BaCl2 Although Kir channel expression was reduced in HC-rat aorta, Kir expression, endothelium-dependent relaxation, and the BaCl2-sensitive component improved in HC rats treated with atorvastatin to reduce serum cholesterol. Remarkably, HC tissues demonstrated increased reactive species (ROS) in smooth muscle cells, which was reversed in rats receiving atorvastatin. In vitro ROS reduction, with superoxide dismutase, improved endothelium-dependent relaxation in HC-rat tissues. Significantly, connexin-43 expression increased in HC aortic tissues, possibly allowing ROS movement into the endothelium and reduction of eNOS activity. In this context, gap junction blockade with 18-β-glycyrrhetinic acid reduced vascular tone in HC rat tissues but not in controls. This reduction was sensitive to NOS inhibition and SOD treatment, possibly as an outcome of reduced ROS influence, and emerged in BaCl2-treated control tissues. In conclusion, our results suggest that early metabolic challenge leads to reduced Kir-mediated endothelium-dependent hyperpolarization, increased vascular ROS potentially impairing NO synthesis and highlight these channels as a possible target for early intervention with vascular dysfunction in metabolic disease. SIGNIFICANCE STATEMENT: The present study examines early endothelial dysfunction in metabolic disease. Our results suggest that reduced inward-rectifier potassium channel function underlies a defective endothelium-mediated relaxation possibly through alteration of nitric oxide synthase activity. This study provides a possible mechanism for the augmentation of relatively small changes in one endothelium-mediated relaxation pathway to affect overall endothelial response and highlights the potential role of inward-rectifier potassium channel function as a therapeutic target to treat vascular dysfunction early in the course of metabolic disease.


[Characteristics of the correction of the blood aggregate condition during extracorporeal hemosorption in hematologic patients].
The authors described experience in the use of the method of extracorporal hemosorption in patients with grave hemoblastoses. Nineteen of them were given 24 hemosorptions. Though severe disorders of the blood aggregate condition, thrombocytopenia (65%) and signs of disorder of thrombocyte function were present, no hemorrhagic complications during hemosorption were observed. Mean doses of heparin taking account of its consumption for sorbent treatment were 200 I.U./kg of body mass in thrombocytopenia and 300 I.U./kg of body mass with the normal level of thrombocytes. The refusal of protamine-sulfate administration to neutralize residual heparin at the end of operation for fear of possible complications made it possible to restore the initial indices of homeostasis 20 hrs. after sorption. This scheme for examination of the homeostasis system made it possible to control the adequacy of patient's heparinization and to avoid complications of the blood coagulation system.


Acceptance of the 2016 Eminent Parasitologist Award: Chemotherapy in the Context of Emerging Parasitic Diseases.
It seems inevitable that chemotherapy will continue, at least in the near-term, to be a significant factor in the control of infectious diseases. The effectiveness of new drugs has invariably been reduced by the emergence of drug resistance. Efforts to thwart resistance should continue, but there are also other areas that might profitably be re-examined. They include (1) methods of new-drug discovery; (2) selection of parasite life-cycle targets; (3) epidemiological determinants of effectiveness in large-scale control programs; and (4) the economic, managerial, psychological, and political ramifications of drug discovery and drug utilization.


Catatonia: "Fluctuat nec mergitur".
In the beginning of the 1900s, the prevalence of catatonia in inpatient samples was reported to be between 19.5% and 50%. From the mid-1900s, most clinicians thought that catatonia was disappearing. Advances in medical sciences, particularly in the field of neurology, may have reduced the incidence of neurological diseases that present with catatonic features or mitigated their severity. More active pharmacological and psychosocial treatment methods may have either eliminated or moderated catatonic phenomena. Moreover, the relatively narrow descriptive features in modern classifications compared with classical texts and ascribing catatonic signs and symptoms to antipsychotic-induced motor symptoms may have contributed to an apparent decline in the incidence of catatonia. The application of catatonia rating scales introduced in the 1990s revealed significantly more symptoms than routine clinical interviews, and within a few years, the notion of the disappearance of catatonia gave way to its un-expected resurgence. Several systematic investigations have found that, on average, 10% of acute psychotic patients present with catatonic features. In this editorial, the changes in the incidence of catatonia and the possible underlying causes are reviewed.


Longitudinal assessment of Integra in primary burn management: a randomized pediatric clinical trial.
Early excision with autograft-allograft closure is standard in severe burn management. Cadaver skin is associated with risks such as antigenicity, infection, and limited availability and shelf life. Previous studies have shown that Integra is safe to use in burns of <20% total body surface area. However, the suitability of its use in large burns (>50% total body surface area), its effects on postburn hypermetabolism, and the long-term cosmetic and functional results have not yet been evaluated. Twenty children with an average burn size of 73 +/- 15% total body surface area (71 +/- 15% full-thickness burn) were randomized to be treated with either Integra or with autograft-allograft technique. Outcome measures such as length of hospital stay, mortality, incidence of infection and sepsis, acute phase protein levels, and muscle fractional synthetic rate were compared between and within groups during the acute stay (admission to discharge). Outcome measures such as resting energy expenditure, body composition data (measured by dual-energy radiograph absorptiometry), cardiac function indexes, and number of reconstructive procedures were compared during acute hospital stay and at long-term follow-up (up to 2 yrs postinjury). Scar evaluation was performed at long-term follow-up. There were no significant differences between Integra and controls in burn size (70 +/- 5% vs. 74 +/- 4% total body surface area), mortality (40% vs. 30%), and length of stay (41 +/- 4 vs. 39 +/- 4 days). In the short term, resting energy expenditure significantly decreased (p < .01), and serum levels of constitutive proteins significantly increased (p < .03) in the Integra group compared with controls. Long-term follow-up revealed a significant increase in bone mineral content and density (24 months postburn, p < .05), as well as improved scarring in terms of height, thickness, vascularity, and pigmentation (12 months and 18-24 months, p < .01) in the Integra group. Integra can be used for immediate wound coverage in children with severe burns without the associated risks of cadaver skin.


Protein malnutrition during pregnancy in C57BL/6J mice results in offspring with altered circadian physiology before obesity.
The mechanisms linking intrauterine growth retardation (IUGR) with adulthood obesity and diabetes are unclear. These studies investigated energy homeostasis in 8- and 20-wk-old male and female mice subjected to protein deficiency in utero. Pregnant C57BL/6J female mice were fed a protein-deficient diet (6% protein). Undernourished offspring (UO) and controls (CO) were cross-fostered to lactating dams fed a 20% control diet. The 24-h profiles of energy expenditure, feeding behavior, physical activity, and whole-body substrate preference was assessed using 8-wk UO and CO weaned onto control diet. Blood chemistries, glucose tolerance, and expression of genes involved in hepatic lipid and glucose metabolism were analyzed in 8- and 20-wk-old CO and UO fed control or a high-fat diet. UO exhibited IUGR with catch-up growth at 8 wk of age and increased severity of diet-induced obesity and insulin resistance by 20 wk of age. Therefore, fetal malnutrition in the C57BL/6J mouse increases sensitivity to diet-induced obesity. Abnormal daily rhythms in food intake and metabolism, increased lipogenesis, and inflammation preceded obesity in the UO group. Arrhythmic expression of circadian oscillator genes was evident in brain, liver, and muscle of UO at 8 and 20 wk of age. Expression of the clock-associated nuclear receptor and transcription repressor Rev-erbalpha was reduced in liver and muscle of UO. Altered circadian physiology may be symptomatic of the metabolic dysregulation associated with IUGR, and altered feeding behavior and substrate metabolism may contribute to the obese phenotype.


YM155, a novel survivin suppressant, enhances taxane-induced apoptosis and tumor regression in a human Calu 6 lung cancer xenograft model.
Survivin, an apoptotic inhibitor, is overexpressed in the majority of human tumor types and represents a novel target for anticancer therapy. Taxanes induce a mitotic cell-cycle block through the inhibition of microtubule depolymerization, with subsequent elevated expression/stabilization of survivin. We investigated the administration of survivin suppressant YM155 monobromide (YM155), in combination with docetaxel, in a human non-small-cell lung cancer (NSCLC) xenograft model. Animals received a 7-day continuous infusion of YM155, 2 mg/kg, and/or three bolus doses of docetaxel, 20 mg/kg, according to three dosing schedules: YM155 administered concomitantly with docetaxel, before docetaxel, and after docetaxel. YM155 administered either concomitantly with or before docetaxel showed significant antitumor activity (tumor regression ≥ 99%), with complete regression of the established human NSCLC-derived tumors in mice (eight of eight and seven of eight animals, respectively). Significantly fewer complete responses (three of eight animals) were achieved when YM155 was administered after docetaxel. No statistically significant decreases in body weight were observed in the combination versus docetaxel groups. YM155 administered concomitantly with docetaxel resulted in significant decreases in mitotic and proliferative indices, and in a significant increase in the apoptosis index. Elevated survivin expression was seen in tumors from mice treated with docetaxel alone; a significant reduction in survivin expression was seen in tumors from mice treated with YM155 alone or in combination with docetaxel, but not in the control group. These results indicate that in a human NSCLC xenograft model YM155 in combination with docetaxel diminished the accumulation of survivin by docetaxel and induced more intense apoptosis and enhanced antitumor activity, compared with single-agent YM155 or docetaxel.


Unsuccessful and Successful Clinical Trials in Acute Respiratory Distress Syndrome: Addressing Physiology-Based Gaps.
The acute respiratory distress syndrome (ARDS) is a severe form of acute hypoxemic respiratory failure caused by an insult to the alveolar-capillary membrane, resulting in a marked reduction of aerated alveoli, increased vascular permeability and subsequent interstitial and alveolar pulmonary edema, reduced lung compliance, increase of physiological dead space, and hypoxemia. Most ARDS patients improve their systemic oxygenation, as assessed by the ratio between arterial partial pressure of oxygen and inspired oxygen fraction, with conventional intensive care and the application of moderate-to-high levels of positive end-expiratory pressure. However, in some patients hypoxemia persisted because the lungs are markedly injured, remaining unresponsive to increasing the inspiratory fraction of oxygen and positive end-expiratory pressure. For decades, mechanical ventilation was the only standard support technique to provide acceptable oxygenation and carbon dioxide removal. Mechanical ventilation provides time for the specific therapy to reverse the disease-causing lung injury and for the recovery of the respiratory function. The adverse effects of mechanical ventilation are direct consequences of the changes in pulmonary airway pressures and intrathoracic volume changes induced by the repetitive mechanical cycles in a diseased lung. In this article, we review 14 major successful and unsuccessful randomized controlled trials conducted in patients with ARDS on a series of techniques to improve oxygenation and ventilation published since 2010. Those trials tested the effects of adjunctive therapies (neuromuscular blocking agents, prone positioning), methods for selecting the optimum positive end-expiratory pressure (after recruitment maneuvers, or guided by esophageal pressure), high-frequency oscillatory ventilation, extracorporeal oxygenation, and pharmacologic immune modulators of the pulmonary and systemic inflammatory responses in patients affected by ARDS. We will briefly comment physiology-based gaps of negative trials and highlight the possible needs to address in future clinical trials in ARDS.


Gallic acid ameliorates behavioral dysfunction, oxidative damage, and neuronal loss in the prefrontal cortex and hippocampus in stressed rats.
Gallic acid (GA) is known to be a natural phenolic compound with antioxidant and neuroprotective effects. This study aims to investigate the impact of GA against restraint stress-induced oxidative damage, anxiety-like behavior, neuronal loss, and spatial learning and memory impairment in male Wistar rats. The animals were divided into four groups (n = 8) and subjected to restraint stress for 4 h per day for 14 consecutive days or left undisturbed (control without inducing stress). In the treatment group, the animals were treated with 2 mL normal saline plus 100 mg/kg GA per day for 14 consecutive days (STR + GA group). The animals received the drug or normal saline by gavage 2 h before inducing restraint stress. ELISA assay measured oxidative stress factors. Elevated-plus maze and Morris water maze tests assessed anxiety-like behavior and spatial learning and memory, respectively. Also, neuronal density was determined using Nissl staining. Restraint stress significantly increased MDA and reduced the activities of GPX and SOD in the stressed rats, which were reserved by treatment with 100 mg/kg GA. Restraint stress markedly enhanced the anxiety-like behavior and spatial learning and memory impairment that were reserved by GA. In addition, treatment with GA reduced the neuronal loss in the stressed rats in the hippocampus and prefrontal cortex (PFC) regions. Taken together, our findings suggest that GA has the potential to be used as a good candidate to attenuate neurobehavioral disorders as well as neuronal loss in the hippocampus and PFC induced by restraint stress via reducing oxidative damage.


Chemotherapy and complications in gynecologic cancer.
Amelioration of toxicity is of major concern to every physician treating patients suffering from gynecologic cancer. If dose escalation in treatment of ovarian cancer is to be considered, the highest priority should be trying to overcome the toxicities of cisplatin, the cornerstone of chemotherapy. So far, dose limiting side-effects are nephro-, neuro-, and ototoxicity. These toxicities may play an even more important role when doses, higher than conventionally given, are to be considered. At higher doses other toxicities such as myelosuppression may become dose limiting. However, nausea and vomiting, which up until now have been intractable, are the most outspoken acute adverse events suffered by the patients. Development of new antiemetic treatment modalities is therefore noteworthy.


Cerebrovascular Imaging: Which Test is Best?
Optimal diagnosis and characterization of cerebrovascular disease requires selection of the appropriate imaging exam for each clinical situation. In this review, we focus on intracranial arterial disease and discuss the techniques in current clinical use for imaging the blood vessel lumen and blood vessel wall, and for mapping cerebral hemodynamic impairment at the tissue level. We then discuss specific strategies for imaging intracranial aneurysms, arteriovenous malformations, dural arterial venous fistulas, and arterial steno-occlusive disease.


Surgical treatment of pulmonary sequestration in adults and children: long-term results.
Few experiences comparing paediatric and adult patients treated for pulmonary sequestration (PS) have been reported. Surgical treatment is considered the best choice, but the time of surgery is still controversial. We present our experience in this setting, comparing characteristics, histological results and outcome of paediatric and adult patients undergoing PS resection. Between 1998 and 2017, a total of 74 patients underwent lobectomy or sublobar resection for PS. Sixty patients were children (group A: ≤16 years old) and 14 were adults (group B: >16 years old). Preoperative diagnosis was radiological. PS was intralobar (42 cases) and extralobar (32 cases). The operation was a muscle-sparing lateral thoracotomy or video-assisted thoracoscopic surgery. Preoperative characteristics, histological results and short-/long-term results of the 2 groups were retrospectively analysed and compared. Thirty-seven percent of the patients in group A presented with respiratory symptoms and 79% in group B (P = 0.44). Most symptomatic patients were treated with a lobectomy. In group A, 2 patients (3%) had a malignant transformation of the lesion. Patients with a prenatal diagnosis treated after the age of 1 year became more symptomatic than those operated on before the age of 1 year (57% vs 23%; P = 0.08). No differences were found in postoperative complications. Long-term stable remission of respiratory symptoms was obtained in 91% of patients in group A and 100% in group B. Adulthood (P = 0.03) and the association with congenital cystic adenomatoid malformation (P = 0.03) were negative prognostic factors for the development of respiratory symptoms. Surgical treatment of PS is safe and feasible. Despite the small number of patients included, study results indicated that an early operation during childhood may prevent the subsequent development of respiratory symptoms. Surgical treatment is also recommended to prevent the rare transformation into malignancy.


Gender disparity in prevalence of depression among patient population: a systematic review.
Many people are unable to withstand the set point for usual vicissitudes of life and are overwhelmed by depression, especially when there is a potential stressor like a disease. Gender is very important in defining susceptibility and exposure to a number of mental health risks. The objective of this review is to systematically identify, appraise and synthesize the best available evidence on gender disparity in prevalence of depression among patient populations. Observational analytical studies done on patients of 18 years old were included. The JBI-MAStARI tool for extraction was used to pool quantitative data. Review Manager Software was used for meta-analysis and Odds ratios and their 95% confidence intervals were calculated. On Meta-analysis, a total of 19639 patients were involved, with male to female ratio of 1.14:1. The finding of the Meta analysis showed that male sex is 63% less likely to develop depression than female sex (Odds ratio=0.63, 95% Confidence Interval= 0.59, 0.68). The studies included were homogenous; Heterogeneity test: Chi(2) = 309.23, df = 30, (P < 0.00001). Depression is more common among females than male patients.


Using particle dimensionality-based modeling to estimate lung carcinogenicity of 3D printer emissions.
The use of 3D printing technologies by industry and consumers is expanding. However, the approaches to assess the risk of lung carcinogenesis from the emissions of 3D printers have not yet been developed. The objective of the study was to demonstrate a methodology for modeling lung cancer risk related to specific exposure levels as derived from an experimental study of 3D printer emissions for various types of filaments (ABS, PLA, and PETG). The emissions of 15 filaments were assessed at varying extrusion temperatures for a total of 23 conditions in a Class 1,000 cleanroom following procedures described by ANSI/CAN/UL 2904. Three approaches were utilized for cancer risk estimation: (a) calculation based on PM2.5 and PM10 concentrations, (b) a proximity assessment based on the pulmonary deposition fraction, and (c) modeling based on the mass-weighted aerodynamic diameter of particles. The combined distribution of emitted particles had the mass median aerodynamic diameter (MMAD) of 0.35 μm, GSD 2.25. The average concentration of PM2.5 was 25.21 μg/m3 . The spline-based function of aerodynamic diameter allowed us to reconstruct the carcinogenic potential of seven types of fine and ultrafine particles (crystalline silica, fine TiO2 , ultrafine TiO2 , ambient PM2.5 and PM10, diesel particulates, and carbon nanotubes) with a correlation of 0.999, P < 0.00001. The central tendency estimation of lung cancer risk for 3D printer emissions was found at the level of 14.74 cases per 10,000 workers in a typical exposure scenario (average cumulative exposure of 0.3 mg/m3 - years), with the lowest risks for PLA filaments, and the highest for PETG type.


Fully Refined Semisynthesis of the [FeFe] Hydrogenase H-Cluster.
[FeFe] hydrogenases contain a 6-Fe cofactor that serves as the active site for efficient redox interconversion between H2 and protons. The biosynthesis of the so-called H-cluster involves unusual enzymatic reactions that synthesize organometallic Fe complexes containing azadithiolate, CO, and CN- ligands. We have previously demonstrated that specific synthetic [Fe(CO)x(CN)y] complexes can be used to functionally replace proposed Fe intermediates in the maturation reaction. Here, we report the results from performing such cluster semisynthesis in the context of a recent fully defined cluster maturation procedure, which eliminates unknown components previously employed from Escherichia coli cell lysate and demonstrate this provides a concise route to H-cluster synthesis. We show that formaldehyde can be used as a simple reagent as the carbon source of the bridging adt ligand of H-cluster in lieu of serine/serine hydroxymethyltransferase. In addition to the actual H-cluster, we observe the formation of several H-cluster-like species, the identities of which are probed by cryogenic photolysis combined with EPR/ENDOR spectroscopy.


Breast-feeding and infant health in a rural US community.
The medical history for the first year of life of 106 infants in a rural community was reviewed. During the first month of life, a significant difference was found in the number of office visits for illness between breast-fed infants and infants who were bottle-fed from birth. For the first 6 months of life, breast-fed infants with and without bottle supplements had significantly fewer months with illness than bottle-fed infants, regardless of prior exposure to breast milk. Demographic data indicated no appreciable differences between feeding groups. It is concluded that breast-feeding offers a protective advantage against illness.


The transport of orthovanadate and similar oxyanions in relation to salt and water transport across the isolated intestine of the common eel, Anguilla anguilla.
An intestinal preparation from the common eel (Anguilla anguilla) was characterized for both mucosal to serosal and serosal to mucosal transport of water, alanine, sodium and chloride. The preparation showed a net outflux of water and a net influx of alanine, and no net fluxes of sodium or chloride. The influxes of all four substances and the outflux of water were inhibited by ouabain, while the outfluxes of alanine, sodium and chloride were increased by ouabain. Influxes and outfluxes of the oxyanions orthovanadate, phosphate, arsenate, sulphate, selenate and chromate were measured over a range of concentrations from 10(-8) M to 10(-4) M. The entry of each anion was dependent on the mucosal concentration, not saturable within the concentration range used or competitive with chloride. There were no net fluxes of these anions and pertechnecate from solutions containing equal mucosal and serosal concentrations of the anion.


Condition dependence and the nature of genetic variation for male sex comb bristle number in Drosophila melanogaster.
Genetic architecture of variation underlying male sex comb bristle number, a rapidly evolving secondary sexual character of Drosophila, was examined. First, in order to test for condition dependence, diet was manipulated in a set of ten Drosophila melanogaster full-sib families. We confirmed heightened condition dependent expression of sex comb bristle number and its female homologue (distal transverse row bristles) as compared to non-sex sternopleural bristles. Significant genotype by environment effects were detected for the sex traits indicating a genetic basis for condition dependence. Next we measured sex comb bristle number and sternopleural bristle number, as well as residual mass, a commonly used condition index, in a set of thirty half-sib families. Sire effect was not significant for sex comb and sternopleural bristle number, and we detected a strong dominance and/or maternal effect or X chromosome effect for both traits. A strong sire effect was detected for condition and its heritability was the highest as compared to sex comb and sternopleural bristles. We discuss our results in light of the rapid response to divergent artificial selection for sex comb bristle number reported previously. The nature of genetic variation for male sex traits continues to be an important unresolved issue in evolutionary biology.


An evaluation of production and economic efficiency of two beef systems from calving to slaughter.
A 3-yr experiment was conducted with cows and their calves to evaluate resource inputs, animal performance, and carcass characteristics of two production systems. In the control system, cows (CON; n = 99/yr) grazed pasture and were fed hay during the winter, and CON steer calves were finished in the feedlot for 211 d after weaning. In the treatment system (TRT; n = 100/yr), cows grazed pasture and crop residue during the winter and were fed hay. Treatment steer calves grazed crop residue after weaning, grazed pasture in the spring and summer, and were finished in the feedlot for 90 d. Body condition scores after TRT cows returned from crop residue grazing were greater (P < 0.01) for CON than for TRT cows. Calving rates were similar for both groups (CON = 91%; TRT = 93%). In the feedlot, CON steers had lower (P < 0.05) ADG and DMI, but were more efficient (P < 0.01) than TRT steers. Treatment steers had greater (P < 0.05) final weight, hot carcass weight and longissimus muscle area, and decreased marbling score. The cost per weaned calf and weaning breakeven were greater (P = 0.07) for the CON system than for the TRT system (CON = 455.12 dollars, 0.91 dollar/0.45 kg; TRT = 421.43 dollars, 0.84 dollar/0.45 kg). When steers were priced into the postweaning phase on an economic basis, slaughter breakeven was lower (P = 0.01), and profit potential tended (P = 0.14) to be greater for TRT steers when they were sold on a live basis. When steers were priced into the postweaning phase on a financial basis, slaughter breakeven was lower (P = 0.03) and profit potential from the sale of steers on a live basis was greater (P = 0.07) for TRT than for CON steers. Economic evaluation of the total system resulted in greater (P = 0.06) profit potential for the TRT system when steers were priced into the system on either an economic or a financial basis and when steers were sold on a live basis, but no differences were observed when steers were sold on a grid basis. Despite differences in cow weight and body condition, calving rates did not differ between systems. Although calves were herdmates, feedlot performance and carcass characteristics differed between systems. The TRT system had lower weaning and slaughter breakeven, lower cost per weaned calf, and greater profit potential when finished steers were sold on a live basis.


Pharmacological Characterization of the Endocannabinoid Sensor GRABeCB2.0.
Introduction: The endocannabinoids (eCBs), 2-arachidonoylglycerol (2-AG) and arachidonoyl ethanolamine (AEA), are produced by separate enzymatic pathways, activate cannabinoid (CB) receptors with distinct pharmacological profiles, and differentially regulate pathophysiological processes. The genetically encoded sensor, GRABeCB2.0, detects real-time changes in eCB levels in cells in culture and preclinical model systems; however, its activation by eCB analogues produced by cells and by phyto-CBs remains uncharacterized, a current limitation when interpreting changes in its response. This information could provide additional utility for the tool in in vivo pharmacology studies of phyto-CB action. Materials and Methods: GRABeCB2.0 was expressed in cultured HEK293 cells. Live cell confocal microscopy and high-throughput fluorescent signal measurements. Results: 2-AG increased GRABeCB2.0 fluorescent signal (EC50=85 nM), and the cannabinoid 1 receptor (CB1R) antagonist, SR141716 (SR1), decreased GRABeCB2.0 signal (IC50=3.3 nM), responses that mirror their known potencies at the CB1R. GRABeCB2.0 fluorescent signal also increased in response to AEA (EC50=815 nM), the eCB analogues 2-linoleoylglycerol and 2-oleoylglycerol (EC50=632 and 868 nM, respectively), Δ9-tetrahydrocannabinol (Δ9-THC), and Δ8-THC (EC50=1.6 and 2.0 μM, respectively), and the artificial CB1R agonist, CP55,940 (CP; EC50=82 nM); however their potencies were less than what has been described at CB1R. Cannabidiol (CBD) did not affect basal GRABeCB2.0 fluorescent signal and yet reduced the 2-AG stimulated GRABeCB2.0 responses (IC50=9.7 nM). Conclusions: 2-AG and SR1 modulate the GRABeCB2.0 fluorescent signal with EC50 values that mirror their potencies at CB1R, whereas AEA, eCB analogues, THC, and CP increase GRABeCB2.0 fluorescent signal with EC50 values significantly lower than their potencies at CB1R. CBD reduces the 2-AG response without affecting basal signal, suggesting that GRABeCB2.0 retains the negative allosteric modulator (NAM) property of CBD at CB1R. This study describes the pharmacological profile of GRABeCB2.0 to improve interpretation of changes in fluorescent signal in response to a series of known eCBs and CB1R ligands.


Dorsal striatopallidal system in anurans.
The dorsal striatopallidal system of tetrapods consists of the dorsal striatum (caudate-putamen in mammals) and the dorsal pallidum. Although the existence of striatal and pallidal structures has been well documented in anuran amphibians, the exact boundaries of these structures have so far been a matter of debate. To delineate precisely the dorsal striatopallidal system of anurans, we used quantitative analysis of leucine-enkephalin immunohistochemistry (in Bombina orientalis, Discoglossus pictus, Xenopus laevis, and Hyla versicolor), retrograde neurobiotin tracing studies (injections in the central and ventromedial thalamic nuclei in H. versicolor), and double-labeling tracing studies (injections in the lateral forebrain bundle and the caudal striatum in B. orientalis). Immunohistochemistry revealed that enkephalin-positive neurons are located mainly in the rostral and intermediate striatum. Neurobiotin tracing studies demonstrated that neurons projecting to the central and ventromedial thalamic nuclei are found in the intermediate and caudal striatum. Double-labeling studies revealed that the population of neurons in the rostral and intermediate striatum innervating the caudal striatum is separated from neurons projecting into the lateral forebrain bundle. Neurons that project to both the caudal striatum and the lateral forebrain bundle are found only in the dorsal part of the intermediate striatum. Taken together, our results suggest that the rostral striatum of anurans is homologous to the striatum proper of mammals, whereas the caudal striatum is comparable to the dorsal pallidum. The intermediate striatum represents a transition area between the two structures.


Obese and lean Zucker rats differ in preferences for sham-fed corn oil or sucrose.
Obese Zucker rats show greater preference for fatty foods in dietary selection studies than do lean rats. We wished to evaluate the preference for dietary fats in Zucker rats in the absence of postingestive cues. Zucker rats (6 lean and 6 obese) were fitted with gastric cannulas for sham feeding. All sham-feeding tests lasted for 30 min. On training days, rats were sham fed 100% corn oil or, on alternate days, rats were sham fed various concentrations of sucrose in volumes yoked to intakes of corn oil. On preference test days, rats were offered 100% corn oil and one sucrose concentration simultaneously for sham feeding. During preference tests, obese rats preferred 100% corn oil to 10% sucrose, whereas lean rats preferred 10% sucrose to 100% corn oil. Obese rats equally preferred 17.5% sucrose and 100% corn oil, whereas lean Zucker rats equally preferred 2.5% sucrose and 100% corn oil. When sucrose concentration was increased to 20% both obese and lean Zucker rats preferred sucrose to 100% corn oil. These results combined with our previously reported results showing that intraduodenally administered fats do not differ in satiating potency for obese and lean Zucker rats (6) strongly suggest that the preference of obese rats for fatty foods is largely mediated by orosensory mechanisms.


Blending polysaccharides with biodegradable polymers. II. Structure and biological response of chitosan/polycaprolactone blends.
Blends of polycaprolactone (PCL) and chitosan (CHT) were prepared by casting from the mixture of solutions of both components in suitable solvents. PCL, and CHT, form phase separated blends with improved mechanical properties and increased water sorption ability with respect to pure PCL. The morphology of the system was investigated by scanning electron microscopy (SEM), atomic force microscopy (AFM) and confocal microscopy. Dispersed domains of CHT in the semicrystalline PCL matrix were found in samples with less than 20% CHT but cocontinuous phase morphologies are found in blends with 20% or more CHT. This feature was corroborated by the temperature dependence of the elastic modulus measured by dynamic mechanical properties as a function of temperature. It was observed that for those blends above 20 wt% CHT, the mechanical stability of the system was kept even after melting of the PCL phase. Primary human chondrocytes were cultured on the different substrates. Cell morphology was studied by SEM and the viability and proliferation was investigated by the colorimetric MTT assay. Different protein conformations were found by AFM on CHT and PCL samples which were related to the biological performance of the substrates. Hydrophilicty of the material is not directly related to the biological response and the sample with 20 wt% CHT shows better results than the other blends with respect to chondrocyte viability and proliferation. However, the results obtained in the blends are worse than in pure PCL. It seems to be correlated with the surface energy of the different blends rather than hydrophilicity.


A workflow for improving estimates of microplastic contamination in marine waters: A case study from North-Western Australia.
Plastic pollution is ubiquitous throughout the marine environment, with microplastic (i.e. <5 mm) contamination a global issue of emerging concern. The lack of universally accepted methods for quantifying microplastic contamination, including consistent application of microscopy, photography, an spectroscopy and photography, may result in unrealistic contamination estimates. Here, we present and apply an analysis workflow tailored to quantifying microplastic contamination in marine waters, incorporating stereomicroscopic visual sorting, microscopic photography and attenuated total reflectance (ATR) Fourier transform infrared (FTIR) spectroscopy. The workflow outlines step-by-step processing and associated decision making, thereby reducing bias in plastic identification and improving confidence in contamination estimates. Specific processing steps include (i) the use of a commercial algorithm-based comparison of particle spectra against an extensive commercially curated spectral library, followed by spectral interpretation to establish the chemical composition, (ii) a comparison against a customised contaminant spectral library to eliminate procedural contaminants, and (iii) final assignment of particles as either natural- or anthropogenic-derived materials, based on chemical type, a compare analysis of each particle against other particle spectra, and physical characteristics of particles. Applying this workflow to 54 tow samples collected in marine waters of North-Western Australia visually identified 248 potential anthropogenic particles. Subsequent ATR-FTIR spectroscopy, chemical assignment and visual re-inspection of photographs established 144 (58%) particles to be of anthropogenic origin. Of the original 248 particles, 97 (39%) were ultimately confirmed to be plastics, with 85 of these (34%) classified as microplastics, demonstrating that over 60% of particles may be misidentified as plastics if visual identification is not complemented by spectroscopy. Combined, this tailored analysis workflow outlines a consistent and sequential process to quantify contamination by microplastics and other anthropogenic microparticles in marine waters. Importantly, its application will contribute to more realistic estimates of microplastic contamination in marine waters, informing both ecological risk assessments and experimental concentrations in effect studies.


Influence of continuous, isobaric rectal distention on smooth muscle tone and cardiovascular activation in healthy volunteers.
Interactions between visceral, intestinal stimulation and autonomic cardiovascular function have received increasing attention over the last years. The aim of the study was to investigate the effect of long-term continuous rectal distention on rectal tone and cardiovascular function at the threshold for first sensation of distention and moderate urge to defecate compared to a resting period without distention. Continuous isobaric rectal distention was performed in 13 healthy volunteers with a ten minute pressure tracking procedure at each subject's predetermined individual threshold for either moderate urge or first sensation, separated by a ten minute resting period. Rectal volume, heart rate, systolic and diastolic blood pressure were determined continuously. Rectal tone decreased significantly over time at the threshold for moderate urge and first sensation. Cardiovascular measures remained stable over the distention procedures, except for a significant increase in systolic blood pressure at the threshold for moderate urge. The previously reported rectal accommodation response to rectal distention was confirmed. The increase in systolic blood pressure is most likely mediated through sympathetic efferent pathways.


Measuring quality in cancer care: overview of initiatives in selected countries.
To inform the Austrian National Cancer Plan on possible generic quality indicators that might be derived from routine data a systematic literature search in three databases, followed by extensive hand-searching to locate initiatives and their publications was carried out in spring 2011. Twenty-one initiatives that developed indicators for measuring quality of cancer care were identified: longer standing and decentralised initiatives are characteristics of the USA. The Canadian province of Ontario publishes the Cancer System Quality Index, centralised audit and peer review programmes are undertaken in the National Health Service in the UK. Methodologically sound cancer type-specific pilot projects in Belgium have been implemented, the Netherlands and Denmark are running national initiatives. Germany recently started quality measurement activities, too. Generic indicators often focus on end-of-life care, multidisciplinarity, advance care planning and documentation. Indicators measuring the quality of care during an entire episode of cancer are rare, as are those for less common cancers and for care in the outpatient setting. Access, equity and the patient's perspective are only beginning to be incorporated into indicators. After having identified a range of candidate indicators that can be implemented with routinely collected data alone, piloting them in Austria would be the next step to go.


Social structure of the harem-forming promiscuous fruit bat, Cynopterus sphinx, is the harem truly important?
Bats are social animals and display a diverse variety of mating and social systems, with most species exhibiting some form of polygyny. Their social organization is fluid and individuals frequently switch partners and roosting sites. While harem-like social organization is observed in multiple tropical species, its importance is contested in many of them. In this study, we investigated the role of harems in the social organization of the old world fruit bat Cynopterus sphinx. Based on regular behavioural observations over a period of 20 months and genetic data from microsatellite markers, we observed that the social organization is flexible, individuals regularly shift between roosts and the social organization resembles a fission-fusion society. Behavioural and genetic analyses suggest that the harems are not strict units of social structure, and the colony does not show signatures of subdivision with harems as behavioural units. We also observed that there was no correlation between individuals with high association index and pairwise relatedness. Our findings indicate that similar to the mating system, the social organization of C. sphinx can also be categorized as a fission-fusion society, and hence the term 'harem' is a misnomer. We conclude that the social system of C. sphinx is flexible, with multi-male multi-female organization, and individuals tend to be loyal to a given area rather than a roost.


Increased Force Required With Proposed Standardized Enteral Feed Connector in Blenderized Tube Feeding.
Enteral tube misconnections resulting in enteral nutrition being provided into the wrong port or tube has been increasingly reported, prompting the development of new international design standards for medical device tubing connectors. Four sample enteral feeds with varying viscosity were placed into a syringe with either the current feeding connector or a prototype ENFit connector. The force (N) required to compress the syringe was gathered through a dynamic mechanical analyzer (RSA G2 Solids Analyzer; TA Instruments). The force needed to compress the syringe was lowest with a fiber-containing formula (Jevity 1 Cal, Abbott Nutrition, Columbus, OH), and a decline in force was detected with the ENFit compared with the current connector (8.61 ± 0.27 N vs 9.62 ± 0.23 N, P < .001). A commercial blenderized formula (Liquid Hope, Functional Formularies, Centerville, OH) required slightly higher force with the ENFit than the current connector (16.82 ± 1.23 N vs 14.5 ± 0.03 N, P < .001). Another commercial blenderized formula (Salmon, Oats, & Squash, Real Food Blends, Chesterton, IN) and a homemade blenderized recipe required significantly more force with the ENFit than the current tube (34.12 ± 0.95 N vs 22.91 ± 0.06 N, P < .001; 34.95 ± 0.06 N vs 27.72 ± 0.07 N, P < .001, respectively). ENFit adapter implementation is the largest change to home enteral nutrition in more than a decade. Additional rigorous testing is required to ensure that the needs of all patients receiving home enteral nutrition are met prior to widespread adoption.


Structural Insight into G Protein-Coupled Receptor Signaling Efficacy and Bias between Gs and β-Arrestin.
G protein-coupled receptors (GPCRs) form the largest family of membrane proteins involved in signal transduction. Because of their ability to regulate a wide range of cellular responses and their dysregulation being associated with many diseases, GPCRs remain a key therapeutic target for several clinical indications. In recent years, it has been demonstrated that ligands for a given receptor can engage distinct pathways with different relative efficacies, a concept known as biased signaling or functional selectivity. However, the structural determinants of this phenomenon remain poorly understood. Using the β2-adrenergic receptor as a model, we identified a linker residue (L1243.43) between the known PIF and NPxxY structural motifs, that plays a central role in the differential efficacy of biased ligands toward the Gs and β-arrestin pathways. Given the high level of conservation of this linker residue, the study provides structural explanations for biased signaling that can be extrapolated to other GPCRs.


Comorbid and co-occurring conditions in migraine and associated risk of increasing headache pain intensity and headache frequency: results of the migraine in America symptoms and treatment (MAST) study.
Migraine has many presumed comorbidities which have rarely been compared between samples with and without migraine. Examining the association between headache pain intensity and monthly headache day (MHD) frequency with migraine comorbidities is novel and adds to our understanding of migraine comorbidity. The MAST Study is a prospective, web-based survey that identified US population samples of persons with migraine (using modified International Classification of Headache Disorders-3 beta criteria) and without migraine. Eligible migraine participants averaged ≥1 MHDs over the prior 3 months. Comorbidities "confirmed by a healthcare professional diagnosis" were endorsed by respondents from a list of 21 common cardiovascular, neurologic, psychiatric, sleep, respiratory, dermatologic, pain and medical comorbidities. Multivariable binary logistic regression calculated odds ratios (OR) and 95% confidence intervals for each condition between the two groups adjusting for sociodemographics. Modeling within the migraine cohort assessed rates of conditions as a function of headache pain intensity, MHD frequency, and their combination. Analyses included 15,133 people with migraine (73.0% women, 77.7% White, mean age 43 years) and 77,453 controls (46.4% women, 76.8% White, mean age 52 years). People with migraine were significantly (P < 0.001) more likely to report insomnia (OR 3.79 [3.6, 4.0]), depression (OR 3.18 [3.0, 3.3]), anxiety (OR 3.18 [3.0 3.3]), gastric ulcers/GI bleeding (OR 3.11 [2.8, 3.5]), angina (OR 2.64 [2.4, 3.0]) and epilepsy (OR 2.33 [2.0, 2.8]), among other conditions. Increasing headache pain intensity was associated with comorbidities related to inflammation (psoriasis, allergy), psychiatric disorders (depression, anxiety) and sleep conditions (insomnia). Increasing MHD frequency was associated with increased risk for nearly all conditions and most prominent among those with comorbid gastric ulcers/GI bleeding, diabetes, anxiety, depression, insomnia, asthma and allergies/hay fever. In regression models controlled for sociodemographic variables, all conditions studied were reported more often by those with migraine. Whether entered into the models separately or together, headache pain intensity and MHD frequency were associated with increased risk for many conditions. Future work is required to understand the causal sequence of relationships (direct causality, reverse causality, shared underlying predisposition), the potential confounding role of healthcare professional consultation and treatment, and potential detection bias.


Alzheimer disease: postmortem neuropathologic correlates of antemortem 1H MR spectroscopy metabolite measurements.
To determine the neuropathologic correlates of antemortem hydrogen 1 ((1)H) magnetic resonance (MR) spectroscopy metabolite measurements in subjects with Alzheimer disease (AD)-type pathology. This study was approved by the institutional review board and was compliant with HIPAA regulations. Informed consent was obtained from each subject. The authors identified 54 subjects who underwent antemortem (1)H MR spectroscopy and were clinically healthy or had AD-type pathology with low to high likelihood of AD according to National Institute on Aging-Reagan neuropathologic criteria at autopsy. They investigated the associations between (1)H MR spectroscopy metabolite measurements and Braak neurofibrillary tangle stage (Braak stage), neuritic plaque score, and AD likelihood, with adjustments for subject age, subject sex, and time between (1)H MR spectroscopy and death. Decreases in N-acetylaspartate-to-creatine ratio, an index of neuronal integrity, and increases in myo-inositol-to-creatine ratio were associated with higher Braak stage, higher neuritic plaque score, and greater likelihood of AD. The N-acetylaspartate-to-myo-inositol ratio proved to be the strongest predictor of the pathologic likelihood of AD. The strongest association observed was that between N-acetylaspartate-to-myo-inositol ratio and Braak stage (R(N)(2) = 0.47, P < .001). Antemortem (1)H MR spectroscopy metabolite changes correlated with AD-type pathology seen at autopsy. The study findings validated (1)H MR spectroscopy metabolite measurements against the neuropathologic criteria for AD, and when combined with prior longitudinal (1)H MR spectroscopy findings, indicate that these measurements could be used as biomarkers for disease progression in clinical trials.


H19 antisense RNA can up-regulate Igf2 transcription by activation of a novel promoter in mouse myoblasts.
It was recently shown that a long non-coding RNA (lncRNA), that we named the 91H RNA (i.e. antisense H19 transcript), is overexpressed in human breast tumours and contributes in trans to the expression of the Insulin-like Growth Factor 2 (IGF2) gene on the paternal chromosome. Our preliminary experiments suggested that an H19 antisense transcript having a similar function may also be conserved in the mouse. In the present work, we further characterise the mouse 91H RNA and, using a genetic complementation approach in H19 KO myoblast cells, we show that ectopic expression of the mouse 91H RNA can up-regulate Igf2 expression in trans despite almost complete unmethylation of the Imprinting-Control Region (ICR). We then demonstrate that this activation occurs at the transcriptional level by activation of a previously unknown Igf2 promoter which displays, in mouse tissues, a preferential mesodermic expression (Pm promoter). Finally, our experiments indicate that a large excess of the H19 transcript can counteract 91H-mediated Igf2 activation. Our work contributes, in conjunction with other recent findings, to open new horizons to our understanding of Igf2 gene regulation and functions of the 91H/H19 RNAs in normal and pathological conditions.


Effects of dibutyryl cyclic adenosine 3':5'-monophosphate on the growth of cultured human small-cell lung carcinoma and the specific cellular activity of L-dopa decarboxylase.
High activity of L-dopa decarboxylase separates small (oat)-cell from non-small-cell lung cancer in cell culture. The present study investigates relationships between the specific cellular activity of this enzyme and: (a) cell growth kinetics of an established line (O-H-1) of human small cell lung carcinoma, and (b) responses of these cells to treatment with cyclic adenosine 3':5'-monophosphate and sodium butyrate. The O-H-1 cells, as for most other established small-cell lines, grow as suspended cell aggregates. During growth, the specific cellular activity of L-dopa decarboxylase parallels levels for [3H]thymidine labeling index and the ratio of cells in G2-M to those in G1-G0 phases of the cell cycle. Each of these parameters is 2- to 3-fold higher during exponential versus stationary growth. Continuous treatment with dibutyryl cyclic adenosine 3':5'-monophosphate (dcAMP; 0.1 or 1 mM) and 1 mM theophylline produces simultaneous cessation of cell growth and an increase in cellular L-dopa decarboxylase activity. During this period, analyses of DNA histograms reveal an increase in the number of cells in the G2-M phase; the rate of increase in the ratio of G2-M to G1-Go cells paralleled the rate of increase in specific activity of the enzyme. The effects of the dcAMP were promptly reversible; release of the apparent G2-M block preceded regrowth of the cells and was accompanied by a return of L-dopa decarboxylase activity to base-line levels. The changes in enzyme activity were specific for cyclic adenosine 3':5'-monophosphate; another cyclic adenosine 3':5'-monophosphate analogue, 8-bromo adenosine cyclic 3':5'-monophosphate yielded similar increases in L-dopa decarboxylase to those seen with dcAMP, while 0.01 to 1 mM butyrate alone produced the inhibition of cell growth but no changes in specific activity of L-dopa decarboxylase or percentage of cells in the different phases of the cell cycle. We conclude that the specific activity of L-dopa decarboxylase, a key neuroendocrine marker for cultured small-cell lung carcinoma, is highest during proliferative growth and/or when these cells are in the G2M phase of the cell cycle. The differential effects of dcAMP and sodium butyrate offer potential for exploring neuroendocrine differentiation in this important lung cancer and related endocrine neoplasms.


Developmental genetics of secretory vesicle acidification during Caenorhabditis elegans spermatogenesis.
Secretory vesicles are used during spermatogenesis to deliver proteins to the cell surface. In Caenorhabditis elegans, secretory membranous organelles (MO) fuse with the plasma membrane to transform spermatids into fertilization-competent spermatozoa. We show that, like the acrosomal vesicle of mammalian sperm, MOs undergo acidification during development. Treatment of spermatids with the V-ATPase inhibitor bafilomycin blocks both MO acidification and formation of functional spermatozoa. There are several spermatogenesis-defective mutants that cause defects in MO morphogenesis, including spe-5. We determined that spe-5, which is on chromosome I, encodes one of two V-ATPase B paralogous subunits. The spe-5 null mutant is viable but sterile because it forms arrested, multi-nucleate spermatocytes. Immunofluorescence with a SPE-5-specific monoclonal antibody shows that SPE-5 expression begins in spermatocytes and is found in all subsequent stages of spermatogenesis. Most SPE-5 is discarded into the residual body during spermatid budding, but a small amount remains in budded spermatids where it localizes to MOs as a discrete dot. The other V-ATPase B subunit is encoded by vha-12, which is located on the X chromosome. Usually, spe-5 mutants are self-sterile in a wild-type vha-12 background. However, an extrachromosomal transgene containing wild-type vha-12 driven by its own promoter allows spe-5 mutant hermaphrodites to produce progeny, indicating that VHA-12 can at least partially substitute for SPE-5. Others have shown that the X chromosome is transcriptionally silent in the male germline, so expression of the autosomally located spe-5 gene ensures that a V-ATPase B subunit is present during spermatogenesis.


Aminosilane micropatterns on hydroxyl-terminated substrates: fabrication and applications.
The technique to pattern aminosilanes on hydroxyl-terminated substrates will open up extensive applications in many fields. There are some existing methods to pattern aminosilanes, in particular, (3-aminopropyl)triethoxysilane (APTES) on SiO(2) and glass substrates through indirect routes. However, few reports focus on the direct patterning of APTES by microcontact printing (microCP), due to the volatility of "inks" which consist of APTES and organic solvents. This report shows that high-quality APTES patterns on hydroxyl-terminated substrates can be directly obtained by microCP using an APTES aqueous solution as "ink". Gold nanoparticles (Au NPs) have been used to verify the presence and quality of APTES patterns on which they are selectively adsorbed. Thus-obtained Au NP patterns can serve as templates for the growth of ZnO nanostructures. Lectins are also successfully immobilized on the APTES patterns, with glutaraldehyde as linker. We believe that our method will serve as a general approach and find a wide range of applications in the fabrication of patterns and devices.


Estimating energy expenditure from raw accelerometry in three types of locomotion.
Accuracy of estimating activity-related energy expenditure (AEE) from raw body acceleration may improve by using prediction equations that are specific for the type of activity. The current study aims to improve published equations by deriving an equation for overground walking and to evaluate whether overground cycling and stair walking require specific prediction equations. Participants (91 male/95 female, 8-81 yr old) were equipped with a triaxial accelerometer (DynaPort MiniMod; McRoberts BV, The Hague, The Netherlands) on their lower back. Total energy expenditure (TEE) was measured using a mobile oxygen analyzer (MetaMax 3b; Cortex Biophysik, Leipzig, Germany). Resting energy expenditure (REE) was measured for 30 min, following which a physical activity course was completed involving walking on level ground at slow (8 min), normal (8 min), and fast speed (3 min), stair walking (3 min), and cycling (8 min). AEE was calculated as TEE - REE, expressed in both absolute (kJ·min) and relative (J·min·kg) units. Mixed linear regression analysis was used for developing regression equations for walking, stair walking, and cycling. Acceleration contributed 76% and 93% (P < 0.001) to explained variance in walking AEE for absolute and relative AEE models, respectively. Age and gender improved estimation accuracy by <1%. Applying a conservative walking equation, AEE (J·min·kg) = -40.19 + 816.11 acceleration (g) (root-mean-square error = 34.00 J·min·kg), to cycling and stair walking resulted in mean bias (95% limits of agreement) of -253 (-449, 46) and -276 (-442, 109) J·min·kg, respectively (approximately 50% bias). Acceleration added 35% and 42% to explained variance in relative AEE (J·min·kg) during cycling and stair walking, respectively; this fraction was approximately 20% for absolute AEE (kJ·min) in both activities. AEE during walking can be predicted across a wide age range using raw acceleration, but activity-specific equations are needed for cycling and stair walking.


Improving information given to patients before endoscopy: a regional audit.
To improve the information given to patients before endoscopy an audit was performed in 16 of 18 endoscopy units in Northern region. Details of current endoscopy information leaflets provided by the 16 respondents were discussed by nurses and consultants from the participating units, and a standard, including 12 separate items, was agreed. Each unit was provided with a comparison of its current leaflet with the standard, which highlighted areas for potential improvement. Six months later the participating units were again asked to provide details of the information; 13 replied, 11 of which had produced new leaflets and two which were in the process of doing so. In the initial survey only 35% (range 8-67%) of the items in the standard were included in the leaflets. Particular omissions were an indication of risks of procedures (three units), notification of follow up procedures (two), details for obtaining the results of the endoscopy (five), advice for people with diabetes (two) and providing a contact number for the endoscopy unit (four). In the repeat audit all 11 units had made changes to their leaflets and, overall, 80% of the items were included. Through this simple audit the range of information given to patients attending for endoscopy in the region has improved.


Significantly increased CD70 up regulation on TEL-AML positive B cell precursor acute lymphoblastic leukemia cells following CD40 stimulation.
TEL-AML the most common genetic alteration in childhood precursor B acute lymphoblastic leukemia (BCP-ALL) is associated with a favorable prognosis. We studied the expression of nerve growth factor/tumor necrosis factor receptor (NGFR/TNFR)/ligand family members on 108 primary BCP-ALL samples by flow cytometry and compared both their baseline expression and CD40-induced modulation on TEL-AML positive and negative leukemia samples. Our findings demonstrate that TEL-AML positive patients exhibit a significantly higher percentage of CD40, CD27 and p75NTR positive blasts at diagnosis. This might well contribute to the improved relapse-free survival of these patients assessed in Kaplan Meier analysis as CD27 and p75NTR directly mediate apoptotic signals. Furthermore CD40 ligation enhances antigen presenting and T cell stimulatory capacity via significant up regulation of CD70 while adequate response to physiological maturation signals as indicated by concomitant down regulation of CD27 is retained in TEL-AML positive leukemia. These data provide novel insights in immunological control mechanisms preserved in this leukemia subtype and suggest that not only treatment with chemicals such as HDAC inhibitors but also retained in vivo response to CD40 ligation contributes to improved immune surveillance in these patients which may add to a superior relapse-free survival observed particularly in the presence of other risk factors.


Homocysteinemia in hypertensive patients with renal target organ damage (mild renal dysfunction).
The prevalence of high plasmatic levels of homocysteine in hypertensive patients with mild renal dysfunction (MRD) defined by 2003 European Hypertension Society Guidelines (men plasmatic creatinine between 1.3 and 1.5; women plasmatic creatinine between 1.2 and 1.4 mg/dl) has not been previously reported. To evaluate this item 18 MRD patients were recruited (54% males, mean age 59.2 +/- 17.3 years, mean plasmatic creatinine 1.30 +/- 0.12 mg/dl). They were compared with a control group of hypertensives with normal renal function (n = 87, 42,9% males, mean age 53.6 +/- 12.3 years, mean plasmatic creatinine 0.83 +/- 0.21 mg/dl) and a group of 29 chronic renal failure patients (51.7% males, mean age 56.9 +/- 15.0 years, mean plasmatic creatinine 2.39 +/- 0.95 mg/dl). Age and sex differences are not significant, plasmatic creatinine levels are different among three groups (p <0.001, t student test). Basal homocysteine levels of CRF (19.3 +/- 7.1 micromol/l) were higher than those of control group (11,0 +/- 4,3 micromol/l) and MRD patients (14.8 +/- 5.5 micromol/l; p = 0.027 vs. CRF and p = 0.007 vs. control, Mann-Whitney test). Mean creatinine clearance was 30.3 +/- 11.5 ml/min for CRF group, significantly lower than MRD patients creatinine clearance (54.5 +/- 9.4 ml/min, p <0.001, t student test) and control ones (88,9 +/- 18,9 ml/min, p <0.001, t student test). Hypertensive patients with mild renal dysfunction showed higher and pathological levels of homocysteinemia as compared with controls, this finding might be related to the higher cardiovascular risk described in this group of patients.


Serotyping of Canadian isolates of Treponema hyodysenteriae and description of two new serotypes.
A total of 30 isolates of Treponema hyodysenteriae collected in the Saint-Hyacinthe (Quebec, Canada) area were serotyped by agar gel double immunodiffusion by using extracted lipopolysaccharide and hyperimmune rabbit antisera. Only 17% (5 of 30) of the isolates were typed with antisera specific for each of the seven known serotypes of T. hyodysenteriae. Antisera raised against 11 untypeable local isolates were then produced and tested against each lipopolysaccharide extract. Results showed two serologically distinct groups among 21 of the 25 untypeable isolates. The isolates in each group shared identical antigens. No detectable reactions could be observed between antisera raised against these 11 isolates and the antigens extracted from 7 reference serotype strains. On the basis of these results, two new serotypes of T. hyodysenteriae, serotypes 8 and 9, are proposed. We also propose isolate FM 88-90 as the reference strain for serotype 8 and isolate FMV 89-3323 as the reference strain for serotype 9. These two new serotypes, which represented 70% of the isolates tested, seem to be the major serotypes found in the province of Quebec.


[A benign variant of the course of Duchenne muscular dystrophy in a child with short stature].
The authors describe a case of a benign variety of progressive Duchenne type muscular dystrophy in a 8-year-old short-stature boy. Provide the electromyographic and electroneuromyographic data, measurements of the growth hormone in blood serum and osseous age. Make suggestions about coupled inheritance of progressive Duchenne type muscular dystrophy and short stature . The latter one is likely to be attended by a decrease of anabolic processes, which may determine a more benign course of the myodystrophic process.