Analysis of mecamylamine stereoisomers on human nicotinic receptor subtypes.
Because mecamylamine, a nicotinic receptor antagonist, is used so often in nicotine research and because mecamylamine may have important therapeutic properties clinically, it is important to fully explore and understand its pharmacology. In the present study, the efficacy and potency of mecamylamine and its stereoisomers were evaluated as inhibitors of human alpha 3 beta 4, alpha 3 beta 2, alpha 7, and alpha 4 beta 2 nicotinic acetylcholine receptors (nAChRs), as well as mouse adult type muscle nAChRs and rat N-methyl-D-aspartate (NMDA) receptors expressed in Xenopus oocytes. The selectivity of mecamylamine for neuronal nAChR was manifested primarily in terms of slow recovery rates from mecamylamine-induced inhibition. Neuronal receptors showed a prolonged inhibition after exposure to low micromolar concentrations of mecamylamine. Muscle-type receptors showed a transient inhibition by similar concentrations of mecamylamine, and NMDA receptors were only transiently inhibited by higher micromolar concentrations. Mecamylamine inhibition of neuronal nAChR was noncompetitive and voltage dependent. Although there was little difference between S-(+)-mecamylamine and R-(-)-mecamylamine in terms of 50% inhibition concentration values for a given receptor subtype, there appeared to be significant differences in the off-rates for the mecamylamine isomers from the receptors. Specifically, S-(+)-mecamylamine appeared to dissociate more slowly from alpha 4 beta 2 and alpha 3 beta 4 receptors than did R-(-)-mecamylamine. In addition, it was found that muscle-type receptors appeared to be somewhat more sensitive to R-(-)-mecamylamine than to S-(+)-mecamylamine. Together, these findings suggest that in chronic (i.e., therapeutic) application, S-(+)-mecamylamine might be preferable to R-(-)-mecamylamine in terms of equilibrium inactivation of neuronal receptors with decreased side effects associated with muscle-type receptors.


Plaque-removal efficacy of four types of dental floss.
Effective plaque removal is essential for gingival health, and dental floss is used to augment plaque removal achieved with a toothbrush. This randomized, controlled, examiner-masked, five-period crossover study examined plaque removal in 25 subjects following single use with an American Dental Association reference manual toothbrush alone and in combination with four floss products: three traditional (unwaxed, woven, and shred-resistant) and one powered flosser. Plaque was scored before and after brushing for 1 minute. The Rustogi modified Navy plaque index was used to focus scores on tooth areas contacted during the proper use of dental floss. Mean plaque reductions (baseline minus postbrushing) in floss contact areas were as follows: 0.181 with the toothbrush alone; 0.228, 0.217, and 0.210 for the toothbrush in combination with the three traditional flosses, unwaxed, woven, and shred-resistant, respectively; and 0.252 for the toothbrush plus powered flosser. No statistically significant differences were found between the three traditional floss treatments. All four floss treatments showed greater (P <0.05) mean plaque removal than the toothbrush alone. Mean plaque removal with the powered flosser combination was greater than for the woven combination and shred-resistant combination (both P < or =0.006) and fell just short of significance compared to the unwaxed combination (P = 0.051). All four floss products in combination with a manual toothbrush removed plaque significantly better than the toothbrush alone. Among floss types, there was evidence of superiority for the powered flosser, but there were no significant treatment differences between the three traditional floss products.


Selecting rep-PCR markers to source track fecal contamination in Laguna Lake, Philippines.
Fecal contamination is one of the factors causing deterioration of Laguna Lake. Although total coliform levels are constantly monitored, no protocol is in place to identify their origin. This can be addressed using the library-dependent microbial source tracking (MST) method, repetitive element sequence-based polymerase chain reaction (rep-PCR) fingerprinting. Serving as a prerequisite in developing the host-origin library, we assessed the discriminatory power of three fingerprinting primers, namely BOX-A1R, (GTG)5, and REP1R-1/2-1. Fingerprint profiles were obtained from 290 thermotolerant Escherichia coli isolated from sewage waters and fecal samples of cows, chickens, and pigs from regions surrounding the lake. Band patterns were converted into binary profiles and were classified using the discriminant analysis of principal components. Results show that: (1) REP1R-1/2-1 has a low genotyping success rate and information content; (2) increasing the library size led to more precise estimates of library accuracy; and (3) combining fingerprint profiles from BOX-A1R and (GTG)5 revealed the best discrimination (average rate of correct classification (ARCC) = 0.82 ± 0.06) in a two-way categorical split; while (4) no significant difference was found between the combined profiles (0.74 ± 0.15) and using solely BOX-A1R (0.76 ± 0.09) in a four-way split. Testing the library by identifying known isolates from a separate dataset has shown that a two-way classification performed better (ARCC = 0.66) than a four-way split (ARCC = 0.29). The library can be developed further by adding more representative isolates per host source. Nevertheless, our results have shown that combining profiles from BOX-A1R and (GTG)5 is recommended in developing the MST library for Laguna Lake.


Exploring nurse preceptors' perceptions of benefits and support of and commitment to the preceptor role in the Western Cape Province.
A preceptor is a specialised tutor who provides practical training to students in the practice setting. They are frequently used to orientate nursing students to prepare them for their duties as professional nurses. In the Western Cape Province professional nurses attend a training programme to prepare them for the role of preceptor. After completion of the training it is unclear how the trained nurse preceptors perceive their preparation for the role. The perceptions of preceptors may influence their commitment to their role. The purpose of this study was to explore nurse preceptors' perceptions of benefits, support and commitment to the preceptor role. A conceptual framework guided the study which replicated previous studies that explored nurse preceptors' perceptions. A quantitative approach utilising a descriptive correlational design was used in this study to address the research questions. A convenience sample was drawn from preceptors (n = 60) who had completed a preceptor training programme at the University of the Western Cape. Instrumentation for the study included the following scales: preceptors' perceptions of benefits and rewards, preceptors' perceptions of support, and commitment to the preceptor role. Data analysis was performed using SPSS 20.0. The findings indicate that nurse preceptors were committed to their role. The findings of this study are supported by Kanter's model of structural determinants of behaviour in organisations, since preceptors who see themselves as having access to opportunity and support are likely to be committed to their role.


Stage II carcinoma of the endometrium: results of therapy and prognostic factors.
A retrospective analysis is reported of 116 patients with Stage II carcinoma of the endometrium treated definitively with combined radiation and total abdominal hysterectomy and bilateral salpingo-oophorectomy (TAH-BSO) or irradiation alone from January 1960 through December 1981. At 5 and 10 years, the overall survival for all patients was 71 and 52% and the disease-free survival was 73 and 69%, respectively. Of 90 patients in the combined therapy group, most received a preoperative intracavitary insertion (3500 mgh to the uterus and 2000 mgh to the upper vagina) and preoperative external beam pelvic irradiation (2000 cGy whole pelvis, additional 3000 cGy to parametria, with midline shield) followed in 4 to 6 weeks by a TAH-BSO. The 5 and 10 year disease-free survival for this group was 78 and 75%, respectively. The incidence of major complications was 7% for the combined therapy group. Twenty-six patients were treated with irradiation alone; most of them received two intracavitary insertions (5000 mgh to the uterus and 3000 mgh to the upper vagina) and external beam pelvic irradiation (2000 cGy whole pelvis, additional 3000 cGy to parametria). The 5 and 10 year disease-free survival was 53 and 45%, respectively. The incidence of major complications was 19%. Factors found to influence the prognosis were histologic grade of tumor, clinical and histologic degree of tumor involvement of the ectocervix, presence of residual tumor in the hysterectomy specimen and the depth of myometrial invasion.


Mind your rhythms: an important role for circadian genes in neuroprotection.
Circadian rhythms govern nearly every physiological process in our brains and bodies. At the most basic level, the molecular clockwork in each cell interacts with metabolic cycles to influence the redox state, allowing for increased cellular activity at specific times of day. In this issue of the JCI, Musiek et al. show that genetic disruptions in the positive arm of the molecular clock can lead to severe astrogliosis, which likely occurs through disruptions in output genes that keep oxidative stress in check. This study demonstrates the importance of proper circadian protein function in the maintenance of neuronal integrity.


On the fate of pumice rafts formed during the 2012 Havre submarine eruption.
Pumice rafts are floating mobile accumulations of low-density pumice clasts generated by silicic volcanic eruptions. Pumice in rafts can drift for years, become waterlogged and sink, or become stranded on shorelines. Here we show that the pumice raft formed by the impressive, deep submarine eruption of the Havre caldera volcano (Southwest Pacific) in July 2012 can be mapped by satellite imagery augmented by sailing crew observations. Far from coastal interference, the eruption produced a single >400 km(2) raft in 1 day, thus initiating a gigantic, high-precision, natural experiment relevant to both modern and prehistoric oceanic surface dispersal dynamics. Observed raft dispersal can be accurately reproduced by simulating drift and dispersal patterns using currents from an eddy-resolving ocean model hindcast. For future eruptions that produce potentially hazardous pumice rafts, our technique allows real-time forecasts of dispersal routes, in addition to inference of ash/pumice deposit distribution in the deep ocean.


Therapeutic switching: from antidermatophytic essential oils to new leishmanicidal products.
This study examined whether the antidermatophytic activity of essential oils (EOs) can be used as an indicator for the discovery of active natural products against Leishmania amazonensis. The aerial parts of seven plants were hydrodistilled. Using broth microdilution techniques, the obtained EOs were tested against three strains of dermatophytes (Trichophyton mentagrophytes, Microsporum gypseum and Microsporum canis). To compare the EOs antifungal and antiparasitic effects, the EOs activities against axenic amastigotes of L. amazonensis were concurrently evaluated. For the most promising EOs, their antileishmanial activities against parasites infecting peritoneal macrophages of BALB/c mice were measured. The most interesting antifungal candidates were the EOs from Cymbopogon citratus, Otacanthus azureus and Protium heptaphyllum, whereas O. azureus, Piper hispidum and P. heptaphyllum EOs exhibited the lowest 50% inhibitory concentration (IC50) values against axenic amastigotes, thus revealing a certain correspondence between both activities. The P. hispidum EO was identified as the most promising product in the results from the infected macrophages model (IC50: 4.7 µg/mL, safety index: 8). The most abundant compounds found in this EO were sesquiterpenes, notably curzerene and furanodiene. Eventually, the evaluation of the antidermatophytic activity of EOs appears to be an efficient method for identifying new potential drugs for the treatment of L. amazonensis.


Biotechnology of extremely thermophilic archaea.
Although the extremely thermophilic archaea (Topt ≥ 70°C) may be the most primitive extant forms of life, they have been studied to a limited extent relative to mesophilic microorganisms. Many of these organisms have unique biochemical and physiological characteristics with important biotechnological implications. These include methanogens that generate methane, fermentative anaerobes that produce hydrogen gas with high efficiency, and acidophiles that can mobilize base, precious and strategic metals from mineral ores. Extremely thermophilic archaea have also been a valuable source of thermoactive, thermostable biocatalysts, but their use as cellular systems has been limited because of the general lack of facile genetics tools. This situation has changed recently, however, thereby providing an important avenue for understanding their metabolic and physiological details and also opening up opportunities for metabolic engineering efforts. Along these lines, extremely thermophilic archaea have recently been engineered to produce a variety of alcohols and industrial chemicals, in some cases incorporating CO2 into the final product. There are barriers and challenges to these organisms reaching their full potential as industrial microorganisms but, if these can be overcome, a new dimension for biotechnology will be forthcoming that strategically exploits biology at high temperatures.


Systematic Analysis of 2'-O-Alkyl Modified Analogs for Enzymatic Synthesis and Their Oligonucleotide Properties.
Enzymatic oligonucleotide synthesis is used for the development of functional oligonucleotides selected by in vitro selection. Expanding available sugar modifications for in vitro selection helps the functional oligonucleotides to be used as therapeutics reagents. We previously developed a KOD DNA polymerase mutant, KOD DGLNK, that enzymatically synthesized fully-LNA- or 2'-O-methyl-modified oligonucleotides. Here, we report a further expansion of the available 2'-O-alkyl-modified nucleotide for enzymatic synthesis by KOD DGLNK. We chemically synthesized five 2'-O-alkyl-5-methyluridine triphosphates and incorporated them into the oligonucleotides. We also enzymatically synthesized a 2'-O-alkyl-modified oligonucleotide with a random region (oligonucleotide libraries). The 2'-O-alkyl-modified oligonucleotide libraries showed high nuclease resistance and a wide range of hydrophobicity. Our synthesized 2'-O-alkyl-modified oligonucleotide libraries provide novel possibilities that can promote the development of functional molecules for therapeutic use.


Splanchnic input to thoracic spinal neurons and its supraspinal modulation in the rat.
The urinary responses of 62 T8-T11 spinal neurons were recorded extracellularly following electrical stimulation of the greater splanchnic nerve (GSN) in chloralose-anesthetized rats. Recorded neurons were found in both the dorsal and ventral horns. Fifty-seven neurons increased their firing rate in response to GSN stimulation; 8 of these exhibited biphasic responses consisting of excitations followed by inhibitions. Excitatory responses to GSN stimulation consisted of either one or two bursts with latencies consistent with activation by either A delta or C fibers. GSN stimulation inhibited 5 neurons. The effects of reversible spinalization on spontaneous activity and on both synchronous and non-synchronous (afterdischarge) GSN-evoked responses were investigated using a cooling probe on the spinal cord between C1 and C2. Of 19 neurons tested in this way, 9 exhibited opposite directional changes in their spontaneous activities and their GSN-evoked responses upon spinalization. Differential effects of cold-block on first and second bursts, or on A delta- and C-fiber mediated responses, were not usually observed. However, differential effects of cold-block on synchronous and non-synchronous portions of the overall GSN-evoked response were often observed in that their magnitudes often changed independently of one another. Supraspinal pathways contributed to GSN-evoked responses of several neurons because their responses were diminished during cooling while spontaneous activity was increased or unchanged. These decreases in the magnitude of the GSN-evoked response were not always accounted for by decreases in the synchronous portions of the responses. However, most neurons did exhibit decreases in the number of non-synchronous responses, or afterdischarges, during spinal cooling, exhibiting in some cases biphasic responses. This study provides evidence for strong supraspinal regulation of splanchnic afferent input to the spinal cord of the rat. Further, this regulation exhibits some specificity toward different portions of splanchnic-evoked responses in spinal neurons.


Common and specific proteins and pathways in heart and cerebral ischemia.
To understand the similarities and differences between acute ischemic stroke and acute myocardial infarction (AMI) to help in the development of specific or common treatment strategies. Using an aptamer-based proteomic array, we measured and compared 1310 circulating proteins in the blood of 40 patients with AIS, 9 patients with AMI, and 31 healthy controls. Pathway enrichment analysis was performed using GSEA and g:profiler. Ninety-four proteins were differentially expressed in AIS, and 284 were differentially expressed in AMI. Of these, 8 were specific to cerebral ischemia, and 197 were specific to myocardial infarction. Forty-two proteins were altered in both ischemia processes. Most altered pathways in AIS could be classified as immune response, cell cycle processing, molecular transport, or signaling. Pathways altered in AMI were mostly related to lipid metabolism and transport, highlighting cholesterol metabolic processes and estrogen signaling. In both types of ischemia, we found pathways related to metabolism, specifically purine metabolism, and signaling processes, such as TNF signaling or MAPK1/3. The present study revealed proteins and pathways that were specifically altered in cerebral ischemia, in cardiac ischemia, or in both diseases, providing information on the similarities and differences of ischemic conditions. The role of common and specific proteins and pathways should be explored in detail to find possible therapeutic targets.


Essential Intracrine Androgenic Action in Lung Development for Both Sexes.
Albeit their recognized negative effects on lung maturation, androgens have been proposed to play an essential positive role in lung development. This work aimed to evaluate the impact of blocking endogenous androgen and estrogen actions and to study the effect of an excess of androgen and estrogen during the end of saccular stage and the beginning of the alveolar stage on lung development. This was performed with normal oxygen atmosphere and with hyperoxia, a model of alveolar simplification, which is observed in new bronchopulmonary dysplasia. Mouse lung samples were collected on postnatal day 9 after exposure to 21% or 80% oxygen (postnatal days 1 to 4), and after administration (postnatal days 3 to 8) of vehicle, pure antiandrogen (flutamide), dihydrotestosterone, pure antiestrogen (fulvestrant), or 17β-estradiol. With 21% oxygen, the major effects on morphometric parameters were induced by flutamide. In contrast, with hyperoxia, both flutamide and dihydrotestosterone had similar effects on several morphometric parameters. For instance, a decrease in the relative frequency of closed areas (mainly composed of saccules/alveoli) < 1000 μm2 and an increase for those > 2500 μm2 were observed after flutamide administration. In conclusion, during the junction between the saccular and the alveolar stages, endogenous androgens play an essential intracrine role in lung development for both sexes while an excess of androgens are deleterious when combined with a hyperoxia treatment, but not with normal oxygen levels. Endogenous estrogens have no effects on the lungs during the developmental window studied, while exogenous estrogens had only isolated effects on some morphometric parameters.


Nuclear receptor atlas of female mouse liver parenchymal, endothelial, and Kupffer cells.
The liver consists of different cell types that together synchronize crucial roles in liver homeostasis. Since nuclear receptors constitute an important class of drug targets that are involved in a wide variety of physiological processes, we have composed the hepatic cell type-specific expression profile of nuclear receptors to uncover the pharmacological potential of liver-enriched nuclear receptors. Parenchymal liver cells (hepatocytes) and liver endothelial and Kupffer cells were isolated from virgin female C57BL/6 wild-type mice using collagenase perfusion and counterflow centrifugal elutriation. The hepatic expression pattern of 49 nuclear receptors was generated by real-time quantitative PCR using the NUclear Receptor Signaling Atlas (NURSA) program resources. Thirty-six nuclear receptors were expressed in total liver. FXR-α, EAR2, LXR-α, HNF4-α, and CAR were the most abundantly expressed nuclear receptors in liver parenchymal cells. In contrast, NUR77, COUP-TFII, LXR-α/β, FXR-α, and EAR2 were the most highly expressed nuclear receptors in endothelial and Kupffer cells. Interestingly, members of orphan receptor COUP-TF family showed a distinct expression pattern. EAR2 was highly and exclusively expressed in parenchymal cells, while COUP-TFII was moderately and exclusively expressed in endothelial and Kupffer cells. Of interest, the orphan receptor TR4 showed a similar expression pattern as the established lipid sensor PPAR-γ. In conclusion, our study provides the most complete quantitative assessment of the nuclear receptor distribution in liver reported to date. Our gene expression catalog suggests that orphan nuclear receptors such as COUP-TFII, EAR2, and TR4 may be of significant importance as novel targets for pharmaceutical interventions in liver.


A randomized controlled trial to test the effectiveness of a peer-based social mobile game intervention to reduce smoking in youth.
Smoking is a major cause of worldwide morbidity and mortality. Almost no evidence-based intervention programs are available to help youth quit smoking. We argue that ineffective targeting of peer influence and engagement difficulties are significant barriers to successful youth smoking cessation. To address these barriers, we developed the mobile game intervention HitnRun. A two-armed randomized controlled trial (RCT; n = 144) was conducted and young smokers (Mage = 19.39; SDage = 2.52) were randomly assigned to either play HitnRun or read a psychoeducational brochure. Prior to, directly following the intervention period, and after three-month follow-up, weekly smoking behavior, abstinence rates, intervention dose, and peer- and engagement-related factors were assessed. Results indicated similar reductions in weekly smoking levels and similar abstinence rates for both groups. Yet, we found a dose effect with HitnRun only: The longer participants played HitnRun, the lower their weekly smoking levels were. In the brochure group, a higher dose was related to higher weekly smoking levels at all measurement moments. Exploratory analyses showed the most powerful effects of HitnRun for participants who connected with and were engaged by the intervention. Future work should build on the promising potential of HitnRun by increasing personalization efforts and strengthening peer influence components.


Hydroxy-α sanshool induces colonic motor activity in rat proximal colon: a possible involvement of KCNK9.
Various colonic motor activities are thought to mediate propulsion and mixing/absorption of colonic content. The Japanese traditional medicine daikenchuto (TU-100), which is widely used for postoperative ileus in Japan, accelerates colonic emptying in healthy humans. Hydroxy-α sanshool (HAS), a readily absorbable active ingredient of TU-100 and a KCNK3/KCNK9/KCNK18 blocker as well as TRPV1/TRPA1 agonist, has been investigated for its effects on colonic motility. Motility was evaluated by intraluminal pressure and video imaging of rat proximal colons in an organ bath. Distribution of KCNKs was investigated by RT-PCR, in situ hybridization, and immunohistochemistry. Current and membrane potential were evaluated with use of recombinant KCNK3- or KCNK9-expressing Xenopus oocytes and Chinese hamster ovary cells. Defecation frequency in rats was measured. HAS dose dependently induced strong propulsive "squeezing" motility, presumably as long-distance contraction (LDC). TRPV1/TRPA1 agonists induced different motility patterns. The effect of HAS was unaltered by TRPV1/TRPA1 antagonists and desensitization. Lidocaine (a nonselective KCNK blocker) and hydroxy-β sanshool (a geometrical isomer of HAS and KCNK3 blocker) also induced colonic motility as a rhythmic propagating ripple (RPR) and a LDC-like motion, respectively. HAS-induced "LDC," but not lidocaine-induced "RPR," was abrogated by a neuroleptic agent tetrodotoxin. KCNK3 and KCNK9 were located mainly in longitudinal smooth muscle cells and in neural cells in the myenteric plexus, respectively. Administration of HAS or TU-100 increased defecation frequency in normal and laparotomy rats. HAS may evoke strong LDC possibly via blockage of the neural KCNK9 channel in the colonic myenteric plexus.


The role of Tregs and CD11c(+) macrophages/dendritic cells in ischemic preconditioning of the kidney.
Dendritic cells have the potential to induce tolerance and here we attempted to identify their role in the tolerance seen in ischemic pre-conditioning. We induced bilateral renal ischemic preconditioning in mice and then challenged them with an ischemic insult 7 days later. Compared to sham-operated controls, preconditioned mice were found to have reduced injury with less inflammation, but had an increased number of regulatory T cells (Tregs) in their kidneys after the delayed insult. Splenocytes from these mice had more Tregs and mature CD11c(+) cells, but reduced proliferative and cytokine-secretory responses, suggesting a state of immunosuppression compared to control mice. Anti-CD25 depletion followed by adoptive transfer of Tregs partially mitigated and then restored the protective effect of preconditioning. Depletion of CD11c(+) cells with liposomes containing clodronate was associated with partial loss of preconditioning benefits. The increased numbers of Tregs or impaired immune response found in splenocytes from preconditioned mice were partially reversed in splenocytes from liposome clodronate-treated animals, suggesting that CD11c(+) cells contribute to immune cell-mediated ischemic preconditioning. Hence, our results show that ischemic preconditioning of the kidney provides a negative signal to the peripheral immune system, partially mediating the tissue-protective and anti-inflammatory effects of this maneuver.


Body composition status and the risk of migraine: A meta-analysis.
To evaluate the association between migraine and body composition status as estimated based on body mass index and WHO physical status categories. Systematic electronic database searches were conducted for relevant studies. Two independent reviewers performed data extraction and quality appraisal. Odds ratios (OR) and confidence intervals (CI) were pooled using a random effects model. Significant values, weighted effect sizes, and tests of homogeneity of variance were calculated. A total of 12 studies, encompassing data from 288,981 unique participants, were included. The age- and sex-adjusted pooled risk of migraine in those with obesity was increased by 27% compared with those of normal weight (odds ratio [OR] 1.27; 95% confidence interval [CI] 1.16-1.37, p < 0.001) and remained increased after multivariate adjustments. Although the age- and sex-adjusted pooled migraine risk was increased in overweight individuals (OR 1.08; 95% CI 1.04, 1.12, p < 0.001), significance was lost after multivariate adjustments. The age- and sex-adjusted pooled risk of migraine in underweight individuals was marginally increased by 13% compared with those of normal weight (OR 1.13; 95% CI 1.02, 1.24, p < 0.001) and remained increased after multivariate adjustments. The current body of evidence shows that the risk of migraine is increased in obese and underweight individuals. Studies are needed to confirm whether interventions that modify obesity status decrease the risk of migraine.


The evaluation of central corneal thickness and intraocular pressure in conjunction with tear IGF-1 levels in patients with acromegaly.
To compare the central corneal thickness (CCT), intraocular pressure (IOP), and tear insulin-like growth factor 1 (IGF-1) levels between patients with acromegaly and a control group and to evaluate the possible effect of tear IGF-1 and duration of the disease on CCT and IOP. We included 31 patients with acromegaly (study group) and 40 age- and sex-matched controls in the study. Patients with acromegaly were divided into 2 subgroups based on disease status (active/inactive). All participants underwent complete ophthalmologic evaluation including CCT and IOP values. Basal tear samples were collected from both groups and tear IGF-1 levels were measured. The CCT, IOP, and tear IGF-1 levels were compared between groups and subgroups and the association between tear IGF-I levels and ocular parameters (CCT, IOP) and disease duration were also evaluated. Central corneal thickness, IOP, and tear IGF-1 levels did not show a significant difference between study and control groups. We also did not find a significant difference in terms of CCT, IOP, or tear IGF-1 levels between subgroups of patients. Correlation analysis did not show an association between the duration of disease and tear IGF-1 levels with CCT or IOP. There was no significant difference in tear IGF-1 levels between patients with acromegaly and controls. Additionally, there was no correlation between disease duration and tear IGF-1 levels with CCT or IOP levels. This lack of association may suggest that tear IGF-1 levels might not have an effect on CCT or IOP findings in patients with acromegaly.


Design of triple-shape memory polyurethane with photo-cross-linking of cinnamon groups.
A triple-shape memory polyurethane (TSMPU) with poly(ε-caprolactone) -diols (PCL-diols) as the soft segments and diphenyl methane diisocyanate (MDI), N,N-bis (2-hydroxyethyl) cinnamamide (BHECA) as the hard segments was synthesized via simple photo-crosslinking of cinnamon groups irradiated under λ > 280 nm ultraviolet (UV) light. Fourier transform infrared spectroscopy (FT-IR), proton nuclear magnetic resonance ((1)H-NMR) and ultraviolet-visible absorption spectrum (UV-vis) confirmed the chemical structure of the material. Differential scanning calorimetry (DSC) and dynamic mechanical analysis (DMA) results demonstrated that the photo-crosslinked polymer possessed two transition temperatures, one is due to the melting point of the soft segment PCL-diols, and the other is due to the glass transition temperature. All these contributed to the cross-linked structure of the hard segments and resulted in an excellent triple-shape memory effect. Alamar blue assay showed that the material has good non-cytotoxicity and can be potentially used in biomaterial devices.


Evidence of extensive lymphocyte death in sheep Peyer's patches. II. The number and fate of newly-formed lymphocytes that emigrate from Peyer's patches.
The emigration of newly produced lymphocytes from Peyer's patches (PP) of lambs was studied. Mesenteric lymph nodes (MLN) were excised from most animals a few weeks after birth, and then at 8 to 10 wk of age, the dividing cells in 3 to 4 m of the small intestine were labeled in situ with [3H]thymidine. An extracorporeal perfusion system was used to restrict the 15-min period of labeling to the perfused lengths of intestine, which included either the large continuous ileal PP or a number of smaller jejunal PP. One or 3 days later, the number of labeled cells in the perfused tissue and in other lymphoid organs was studied by autoradiography. In the perfused tissues, labeled lymphocytes accounted for 63.7% of ileal PP cells by 1 day and for 86.7% by 3 days compared with only 9.6% of lymphocytes in the perfused MLN. Labeled lymphoid cells in the perfused PP were nearly all in the follicles. Labeled lymphocytes that must have been produced in the segments of ileum or jejunum at the time of the perfusion, subsequently emigrated via the lymphatics, and were identified in the spleen, MLN, other lymph nodes, blood, jejunal PP, and at a lower frequency in the thymus, nonperfused ileal PP, and bone marrow. In lymph nodes, spleen, and nonperfused PP, more than 80% of the immigrant newly formed PP-derived cells were small- and medium-sized lymphocytes, and about 15% were large lymphocytes. The nature of the labeled cells in the lamina propria of the nonperfused small intestine was quite different in that approximately 50% were plasma cells as early as 24 hr after the cells were born in the perfused gut. It is proposed that terminal B cell differentiation was most likely initiated within the PP in response to the entry of antigen. It was estimated that at both 1 and 3 days after perfusion there were about 100 times more labeled cells in the perfused ileal PP than could be accounted for by emigration to other organs. It was concluded that these results provide additional support for the view that PP in lambs produce a tremendous number of lymphocytes, but relatively few leave their site of production; most apparently die in situ.


Factors for graft-versus-host disease after donor lymphocyte infusions with an escalating dose regimen: lack of association with cell dose.
We investigated the risk factors for graft-versus-host disease (GVHD) in 82 patients treated with donor lymphocyte infusions (DLI) using an escalating dose regimen for chronic myeloid leukaemia in relapse following conventional allografting. Two factors emerged as predictors of both acute and chronic GVHD: the infusion of male recipients with lymphocytes from a female donor and the interval between transplant and last DLI, but only the first remained significant at multivariate analysis. Surprisingly, lymphocyte dose did not influence the incidence of GVHD. Our results suggest that DLI can be given in large cell doses without increasing the risk of GVHD.


Barium borohydride chlorides: synthesis, crystal structures and thermal properties.
Here we report the synthesis, mechanism of formation, characterization and thermal decomposition of new barium borohydride chlorides prepared by mechanochemistry and thermal treatment of MBH4-BaCl2, M = Li, Na or K in ratios 1 : 1 and 1 : 2. Initially, orthorhombic barium chloride, o-BaCl2 transforms into o-Ba(BH4)xCl2-x, x ∼ 0.15. Excess LiBH4 leads to continued anion substitution and a phase transformation into hexagonal barium borohydride chloride h-Ba(BH4)xCl2-x, which accommodates higher amounts of borohydride, possibly x ∼ 0.85 and resembles h-BaCl2. Thus, two solid solutions are in equilibrium during mechano-chemical treatment of LiBH4-BaCl2 (1 : 1) whereas LiBH4-BaCl2 (2 : 1) converts to h-Ba(BH4)0.85Cl1.15. Upon thermal treatment at T > ∼200 °C, h-Ba(BH4)0.85Cl1.15 transforms into another orthorhombic barium borohydride chloride compound, o-Ba(BH4)0.85Cl1.15, which is structurally similar to o-BaBr2. The samples with M = Na and K have lower reactivity and form o-Ba(BH4)xCl2-x, x ∼ 0.1 and a solid solution of sodium chloride dissolved in solid sodium borohydride, Na(BH4)1-xClx, x = 0.07. The new compounds and reaction mechanisms are investigated by in situ synchrotron radiation powder X-ray diffraction (SR-PXD), Fourier transform infrared spectroscopy (FT-IR) and simultaneous thermogravimetric analysis (TGA), differential scanning calorimetry (DSC), mass spectroscopy (MS) and temperature programmed photographic analysis (TPPA).


Understanding non-coding DNA regions in yeast.
Non-coding transcripts play an important role in gene expression regulation in all species, including budding and fission yeast. Such regulatory transcripts include intergenic ncRNA (non-coding RNA), 5' and 3' UTRs, introns and antisense transcripts. In the present review, we discuss advantages and limitations of recently developed sequencing techniques, such as ESTs, DNA microarrays, RNA-Seq (RNA sequencing), DRS (direct RNA sequencing) and TIF-Seq (transcript isoform sequencing). We provide an overview of methods applied in yeast and how each of them has contributed to our knowledge of gene expression regulation and transcription.


Flavonoid-Coated Gold Nanoparticles as Efficient Antibiotics against Gram-Negative Bacteria-Evidence from In Silico-Supported In Vitro Studies.
Flavonoids are a class of bioactive plant-derived natural products that exhibit a broad range of biological activities, including antibacterial ones. Their inhibitory activity toward Gram-positive bacterial was found to be superior to that against Gram-negative ones. In the present study, a number of flavonoid-coated gold nanoparticles (GNPs) were designed to enhance the antibacterial effects of chrysin, kaempferol, and quercetin against a number of Gram-negative bacteria. The prepared GNPs were able to conjugate to these three flavonoids with conjugation efficiency ranging from 41% to 80%. Additionally, they were able to exert an enhanced antibacterial activity in comparison with the free flavonoids and the unconjugated GNPs. Quercetin-coated GNPs were the most active nano-conjugates and were able to penetrate the cell wall of E. coli. A number of in silico experiments were carried out to explain the conjugation efficiency and the antibacterial mechanisms of these flavonoids as follows: (i) these flavonoids can efficiently bind to the glutathione linker on the surface of GNPs via H-bonding; (ii) these flavonoids, particularly quercetin, were able to increase the bacterial membrane rigidity, and hence decrease its functionality; (iii) these flavonoids can inhibit E. coli's DNA gyrase (Gyr-B) with IC50 values ranging from 0.9 to 3.9 µM. In conclusion, these bioactive flavonoid-based GNPs are considered to be very promising antibiotic candidates for further development and evaluation.


Inhibition of a protein tyrosine phosphatase using mesoporous oxides.
The feasibility of utilizing mesoporous matrices of alumina and silica for the inhibition of enzymatic activity is presented here. These studies were performed on a protein tyrosine phosphatase by the name chick retinal tyrosine phosphotase-2 (CRYP-2), a protein that is identical in sequence to the human glomerular epithelial protein-1 and involved in hepatic carcinoma. The inhibition of CRYP-2 is of tremendous therapeutic importance. Inhibition of catalytic activity was examined using the sustained delivery of p-nitrocatechol sulfate (pNCS) from bare and amine functionalized mesoporous silica (MCM-48) and mesoporous alumina (Al(2)O(3)). Among the various mesoporous matrices employed, amine functionalized MCM-48 exhibited the best release of pNCS and also inhibition of CRYP-2. The maximum speed of reaction v(max) (=160 +/- 10 micromol/mnt/mg) and inhibition constant K(i) (=85.0 +/- 5.0 micromol) estimated using a competitive inhibition model were found to be very similar to inhibition activities of protein tyrosine phosphatases using other methods.


Key findings from a prospective trauma registry at a regional hospital in Southwest Cameroon.
Trauma is a leading cause of morbidity and mortality worldwide. Data characterizing the burden of trauma in Cameroon is limited. Regular, prospective injury surveillance can address the shortcomings of existing hospital administrative logs and medical records. This study aims to characterize trauma as seen at the emergency department (ED) of Limbe Regional Hospital (LRH) and assess the completeness of data obtained by a trauma registry. From January 2008 to October 2013, we prospectively captured data on injured patients using a strategically designed, context-relevant trauma registry instrument. Indicators around patient demographics, injury characteristics, delays in accessing care, and treatment outcomes were recorded. Descriptive, bivariate, and multivariate statistical analyses were conducted. About 5,617 patients, aged from 0.5-95years (median age of 26 years), visited the LRH ED with an injury; 67% were male. Students (27%) were the most affected occupation category. Road traffic injuries (RTIs) (56%), assault (22%), and domestic injuries (13%) were the leading causes of injury. Two-thirds of RTIs were motorcycle-related. Working in transportation (AOR 4.42, p<0.001) and law enforcement (AOR 1.73, p = 0.004) were significant predictors of having a RTI. The trauma registry showed a significant improvement in completeness of all data (p<0.001) and it improved over time compared with previous administrative records. However, proportions of missing data still ranged from 0.5% to 8.2% and involved respiratory rate or Glasgow Coma scale. Implementation of a context-appropriate trauma registry in resource-constrained settings is feasible. Providing valuable, high-quality data, the trauma registry can inform trauma care quality improvement efforts and policy development. Study findings indicate the need for injury prevention interventions and policies that will prioritize high-risks groups, such as those aged 20-29 years, and those in occupations requiring frequent road travel. The high incidence of motorcycle-related injuries is concerning and calls for a proactive solution.


Risk for endometrial cancer in relation to occupational physical activity: a nationwide cohort study in Sweden.
Notwithstanding its biologic plausibility, the association between physical activity and endometrial cancer has been analyzed in only a few epidemiological studies. Retrospective assessment of exposure and small sample size often hampers interpretation of published data. We studied risk for endometrial cancer in relation to physical activity at work in a large cohort of Swedish women identified in the nationwide censuses in 1960 and 1970, with jobs that could be consistently classified into one of 4 levels of physical demands. Follow-up from 1971 through 1989 was accomplished through record linkages. Multivariate Poisson regression models were used to estimate relative risk. The risk for endometrial cancer increased regularly with decreasing level of occupational physical activity (p for trend < 0.001), and was associated more strongly with activity in 1970 than in 1960. In multivariate analyses, adjusted for age at follow-up, place of residence, calendar year of follow-up, and social class, the relative risk among women with the same physical activity level in 1960 and in 1970 was 30% higher for sedentary as compared with high/very high activity level; (p for trend=0.04). The protective effect of physical activity appeared to be confined to women aged 50 to 69, among whom sedentary work was associated with a 60% higher risk than that observed among women estimated to be physically most active. The excess seemed to disappear within 10 years after a change in physical activity level. Although confounding cannot be ruled out in our data, occupational physical activity appears to reduce the risk for endometrial cancer.


Numerical simulation of a BPH thermal therapy--a case study involving TUMT.
The use of numerical simulation as a means to predict the outcome of transurethral microwave thermotherapy (TUMT) is set forth in detail. The simulation was carried out as a case study of a specific TUMT procedure. The selection of the case study was based on the availability of extensive medical records which documented an extraordinary application of TUMT. Predictions were made of the time-varying temperature patterns within the prostate, the bladder, the sphincter, the pelvic floor, and the fat and connective tissue which envelop these organs. These temperature patterns provided the basis of maps which highlighted those locations where necrosis occurred. An injury integral was used to predict the extent of the necrotic tissue produced by the therapy. It was found that, for the specific case being considered, necrosis occurred not only within the prostate but also extended to the neck of the bladder and to the fatty tissue. A special feature of the simulation was the accounting of the liquid-to-vapor phase change of the interstitial water. The vapor generated by the phase change is believed to significantly enlarge the region of necrosis. By the same token, the vapor pressure is expected to cause motion of the high-temperature liquid to deep-tissue regions. The damage predicted by the numerical simulation was compared, in detail, with post-operative medical examinations and found to be corroborated.


Evolutionary principles of modular gene regulation in yeasts.
Divergence in gene regulation can play a major role in evolution. Here, we used a phylogenetic framework to measure mRNA profiles in 15 yeast species from the phylum Ascomycota and reconstruct the evolution of their modular regulatory programs along a time course of growth on glucose over 300 million years [corrected]. We found that modules have diverged proportionally to phylogenetic distance, with prominent changes in gene regulation accompanying changes in lifestyle and ploidy, especially in carbon metabolism. Paralogs have significantly contributed to regulatory divergence, typically within a very short window from their duplication. Paralogs from a whole genome duplication (WGD) event have a uniquely substantial contribution that extends over a longer span. Similar patterns occur when considering the evolution of the heat shock regulatory program measured in eight of the species, suggesting that these are general evolutionary principles. DOI:http://dx.doi.org/10.7554/eLife.00603.001.


Evaluation of antioxidant and inhibitory activities for different subclasses flavonoids on enzymes for rheumatoid arthritis.
Antioxidant activities of flavonoids were decreased in the order of flavonols > flavanones > flavones. Inhibitory intensities for hyaluronidase and collagenase reaction differed clearly according to flavonoid subclasses. Kaempferol, quercetin, myricetin, and rutin in flavonols inhibited hyaluronidase reaction specifically, while apigenin, luteolin, baicalin, and baicalein in flavones showed specific inhibition to collagenase reaction. In addition, the flavonoids, except baicalin and catechin, inhibited potently LPS-induced nitrite production in a dose-dependent manner, which might be mainly due to the suppression of inducible nitric oxide (NO) synthase. Quercetin and luteolin showed the strongest inhibitory activities on 15-lipoxygenase (LOX), and quercetin showed relatively potent inhibition on cyclooxygenase-1 (COX-1) reaction. Otherwise, all tested flavonoids possessed the inhibitory activity to COX-2 reaction, and especially luteolin, kaempferol, hesperetin, and naringin showed relatively the potent inhibition on COX-2 reaction. This report elucidated the anti-inflammatory activities, such as the antioxidant property, inhibition of NO production, and inhibition of inflammatory enzymes (hyaluronidase, collagenase, LOX, and COXs) of several subclass flavonoids.


Neoantigen-Based Nanovaccine In Combination with Immune Checkpoint Inhibitors Abolish Postsurgical Tumor Recurrence and Metastasis.
The notorious limitation of conventional surgical excision of primary tumor is the omission of residual and occult tumor cells, which often progress to recurrence and metastasis, leading to clinical treatment failure. The therapeutic vaccine is emerging as a promising candidate for dealing with the issue of postsurgical tumor residuals or nascent metastasis. Here, a flexible and modularized nanovaccine scaffold based on the SpyCatcher003-decorated shell (S) domain of norovirus (Nov) is employed to support the presentation of varied tumor neoantigens fused with SpyTag003. The prepared tumor neoantigen-based nanovaccines (Neo-NVs) are able to efficiently target to lymph nodes and engage with DCs in LNs, triggering strong antigen-specific T-cell immunity and significantly inhibiting the growth of established orthotopic 4T1 breast tumor in mice. Further, the combination of Neo-NVs and anti-PD-1 monoclonal antibody (mAb) produces significant inhibition on postsurgical tumor recurrence and metastasis and induces a long-lasting immune memory. In conclusion, the study provides a simple and reliable strategy for rapid preparing personalized neoantigens-based cancer vaccines and engaging checkpoint treatment to restore the capability of tumor immune surveillance and clearance in surgical patients.


The assessment of total antioxidant capacity and superoxide dismutase levels, and the possible role of manganese superoxide dismutase polymorphism in acromegaly.
Oxidative status is attributed to endothelial dysfunction and might be one of the key mechanisms of endothelial dysfunction in acromegaly. In this study, we aimed to investigate the effect of acromegaly on superoxide dismutase (SOD) and total antioxidant capacity (TAC) levels, and the possible influence of human manganese superoxide dismutase (MnSOD) polymorphism on these levels. 51 acromegaly patients and 57 age and sex matched healthy subjects were recruited to the study in Bezmialem Vakif University Hospital between 2011 and 2014. The median SOD and TAC levels were 42.7 (33-60) pg/mL and 1,313.7 (155-1,902) μM in acromegaly; and 46.3 (38-95) pg/mL and 1,607.3 (195-1,981) μM in healthy subjects (p < 0.001, p < 0.001). SOD levels were decreased in controlled and uncontrolled patients compared to healthy subjects (p = 0.05 and p = 0.002, respectively). Controlled and uncontrolled acromegaly displayed significantly decreased levels of TAC compared to healthy subjects (p < 0.05 and p < 0.001, respectively). SOD levels were not associated with MnSOD polymorphisms in acromegaly. In conclusion, this study showed that acromegaly was associated with decreased levels of SOD and TAC, and controlling the disease activity could not adequately improve these levels.


Ocrelizumab-induced inflammatory bowel disease-like illness characterized by esophagitis and colitis.
Ocrelizumab is an intravenous anti-CD20 monoclonal antibody, approved for use in primary progressive multiple sclerosis due to its selective depletion of B-lymphocytes. Herein we describe the case of a 56-year-old female who developed odynophagia and bloody diarrhea following treatment with ocrelizumab. This was characterized endoscopically by ulcerations in the esophagus and colon. The patient was treated with high-dose intravenous glucocorticoids with good clinical response.


The impact of gynaecological cancer treatment on physical activity levels: a systematic review of observational studies.
The natural history of physical activity levels during and following gynaecological cancer treatment is not well understood. This is required in order to establish the time at which physical activity levels are lowest in order to target cancer rehabilitation or exercise interventions in gynaecological cancer population accordingly. To conduct a systematic review to evaluate the impact of gynaecological cancer treatments on physical activity levels and to summarise the pattern of changes in physical activity levels over time among patients with gynaecological cancer. A comprehensive literature search was performed via MEDLINE (1946-2018), CINAHL (1982-2018), EMBASE (1947-2018), Ovid Emcare (1947-2018), PsycINFO (1806-2018) and the Cochrane Library (1991-2018). Studies were eligible for inclusion if they had assessed changes in physical activity levels during and after gynaecological cancer treatment. The methodological quality of the eligible studies was assessed by two independent reviewers using the Joanna Briggs Institute Critical Appraisal Tools. In total, six studies (three cohort studies and three cross-sectional studies) with 1607 participants were included. All studies used patient-reported physical activity measures. Two of the three cohort studies measured patient-recalled physical activity levels before diagnosis (baseline), and length of follow-up varied across all studies. The majority of participants were treated surgically±adjuvant therapy. Physical activity levels decreased at 6 months following surgery when compared with pre-treatment levels. Approximately 91% of participants did not meet physical activity guidelines 2 years following diagnosis, and 58% reported being less physically active 3 years after diagnosis, compared with the pre-diagnosis levels. Despite the paucity of evidence and limitations in the current body of literature, this review demonstrated that compared to pre-diagnosis, levels of physical activity remain low in gynaecological cancer survivors up to 3 years after diagnosis. More research is warranted to better characterise the pattern of change of physical activity levels across the disease trajectory and identify changes in physical activity patterns by cancer treatments and gynaecological tumour streams in order to target interventions accordingly.


A clathrin-dynamin-dependent endocytic pathway for the uptake of HIV-1 by direct T cell-T cell transmission.
Cellular contacts between HIV-1-infected cells and target primary T CD4+ lymphocytes trigger the formation of a structure known as the virological synapse. As a consequence, viral production in HIV-1-infected cells is polarized towards the virological synapse and nascent viral particles are directly transferred to target T CD4+ lymphocytes. In this study, we performed short time cocultures of target primary T CD4+ lymphocytes with effector T cells infected by either HIV-1 NL4-3 or BaL. Using flow cytometry and immuno-confocal analyses, we investigated the transfer of HIV-1 virion antigens. We found that after 3 h of coculture, unstimulated T CD4+ lymphocytes captured complete HIV-1 virions from infected T cells during cell-cell contacts. Virus transfer occurred through a dynamin-dependent pathway and could be inhibited by chlorpromazine, an inhibitor of clathrin-dependent endocytosis. Transferred HIV-1 virions were located in compartments close to the surface of the target cell in a polarized manner. These compartments were positive for clathrin and the early endosomal marker EEA1 but were negative for caveolin-1. Furthermore, the great majority of internalized HIV-1 particles did not colocalize with Lamp1, a well-known marker for the lysosomal-degradative pathway. Similar results were observed when stimulated primary T CD4+ lymphocytes were the target cells. Our results suggest a mechanism of cell to cell HIV-1 transfer through a clathrin- and dynamin-dependent early endocytic pathway where internalized HIV-1 particles would not reach Lamp1 positive compartments, suggesting that during HIV-1 transfer by cell-cell contacts, virions can be taken up by endocytosis but not be degraded in lysosomes.


The management of gingival recession.
This, the second paper in a short series on gingival recession, discusses the various modalities of treatment for gingival recession and the indications for treatment. Special consideration is given to the surgical treatment techniques. Factors affecting the outcome of treatment and success rates of the treatment options are also discussed.


Age-related expression analysis of mouse liver nuclear protein binding to 3'-untranslated region of Period2 gene.
In mammals, both circadian rhythm and aging play important roles in regulating time-dependent homeostasis. We previously discovered an age-related increase element binding protein, hnRNP A3, which binds to the 3'-untranslated region (UTR) of blood coagulation factor IX (FIX). Here, we describe other members of this protein family, hnRNP C and hnRNP H, which bind to the 3'-UTR of the mouse circadian clock gene Period 2 (mPer2). RNA electrophoretic mobility shift assays using a (32)P-labeled Per2 RNA probe coupled with two-dimensional gel electrophoresis followed by MALDI-TOF/MS peptide mass fingerprint analysis was used to analyze these proteins. Western blotting suggested that the total expression of these proteins in mouse liver cell nuclei does not increase with age. Two-dimensional gel electrophoresis analysis of age-related protein expression showed that many isoforms of these proteins exist in the liver and that each protein exhibits a complex age-related expression pattern. These results suggest that many isoforms of proteins are regulated by different aging systems and that many age regulation systems function in the liver.


Immune cytokine regulation of sympathetic ganglion response to injury.
Without appropriate response to traumatic injury, the nervous system cannot survive. The superior cervical (sympathetic) ganglion of the neonatal rat has been a uniquely valuable model system in which to study neuronal response to traumatic injury. Explantation of the ganglion into culture involves severing both presynaptic fibers (compromising preganglionic input) and axotomy (damaging efferent axons of ganglionic neurons). Since traumatic injury of any kind involves the elaboration of inflammatory cytokines, we have been interested in examining the action of these inflammatory cytokines on neurotransmitter gene expression in injured sympathetic ganglia.


Bacillus infernus sp. nov., an Fe(III)- and Mn(IV)-reducing anaerobe from the deep terrestrial subsurface.
Bacillus infernus sp. nov. was isolated from ca. 2,700 m below the land surface in the Taylorsville Triassic Basin in Virginia. B. infernus was a strict anaerobe that grew on formate or lactate with Fe(III), MnO2, trimethylamine oxide, or nitrate (reduced to nitrite) as an electron acceptor, and it also grew fermentatively on glucose. Type strain TH-23 and five reference strains were gram-positive rods that were thermophilic (growth occurred at 61 degrees C), halotolerant (good growth occurred in the presence of Na+ concentrations up to 0.6 M), and very slightly alkaliphilic (good growth occurred at pH 7.3 to 7.8). A phylogenetic analysis of its 16S rRNA indicated that B. infernus should be classified as a new species of the genus Bacillus. B. infernus is the only strictly anaerobic species in the genus Bacillus.


Anti-proteinase 3 antineutrophil cytoplasmic antibody reflects disease activity and predicts the response to steroid therapy in ulcerative colitis.
Serum anti-proteinase 3 antineutrophil cytoplasmic antibody (PR3-ANCA) is a disease-specific antibody against granulomatosis with polyangiitis. PR3-ANCA is a useful serological marker for disease severity in ulcerative colitis (UC). The purpose of this study was to investigate whether PR3-ANCA levels could also predict the success of induction therapy and to compare its performance against other markers, including serum CRP and fecal hemoglobin. This was a multicenter retrospective study. In total, 159 patients with active-phase UC underwent colonoscopy. Disease activity was measured using the Mayo endoscopic subscore (MES). PR3-ANCA positivity and the response to induction therapy, either 5-aminosalicylic acid or steroid, were assessed. PR3-ANCA, CRP, and fecal hemoglobin were measured during the active phase, and during clinical remission. Eighty-five (53.5%) of 159 patients with active UC were positive for PR3-ANCA. PR3-ANCA titers were significantly higher in the group of patients with MES 3 compared to patients with MES 1 (P = 0.002) or MES 2 (P = 0.035). Steroid therapy was administered to 56 patients with a median partial Mayo score of 7 (5-9), which is equivalent to moderate-to-severe disease activity. PR3-ANCA positivity of non-responders to steroid therapy was significantly higher than that of responders (71.9% vs, 41.7%, P = 0.030), whereas CRP and fecal hemoglobin were not predictive of steroid response. Multivariate analysis demonstrated that PR3-ANCA positivity was associated with non-response to steroid therapy (odds ratio 5.19; 95% confidence interval, 1.54-17.5; P = 0.008). Of the 37 patients treated to clinical remission who were also positive for PR3-ANCA during the active phase, 27 had an MES of ≥ 1, and 10 patients had an MES of 0. In clinical remission, the proportion of patients with MES 0 in 17 patients whose PR3-ANCA became negative was significantly higher than that in 20 patients whose PR3-ANCA remained positive (47.1% vs. 10.0%, P = 0.023). PR3-ANCA not only serves as a marker of disease activity, but also predicts the failure of steroid therapy in moderate-to-severe UC. This study was retrospectively registered in the UMIN Clinical Trials Registry System (000039174) on January 16, 2020.


A combined phylogenetic strategy illuminates the evolution of Goniodorididae nudibranchs (Mollusca, Gastropoda, Heterobranchia).
Goniodorididae is a family of small dorid nudibranchs distributed worldwide that feed on entoprocts, ascidians, and bryozoans. The evolutionary relationships between its taxa have been uncertain due to the limited taxa available for phylogenetic analyses; some genera being paraphyletic. The family includes a remarkable number of synonymized genera in which the species richness is unequally distributed, while some genera have dozens of species others are monospecific. Some clades are very uniform morphologically while others are considered highly variable. To increase backbone phylogenetic resolution a target enrichment approach of ultra-conserved elements was aimed at representative Goniodorididae species for the first time. Additionally, we increase species representation by including mitochondrial markers cytochrome c oxidase subunit I and ribosomal RNA 16S as well as nuclear Histone 3 and ribosomal RNA 18S from 109 Goniodorididae species, out of approximately 160 currently valid species. Maximum likelihood and Bayesian inference analyses were performed to infer the phylogeny of the family. As a result, two subfamilies and eleven genera were elucidated. The synonymized genera Bermudella, Cargoa, and Ceratodoris are here resurrected and a new genus, Naisdoris gen. nov., is described. The clades included taxa with shared prey preference, showing that trophic behavior could have driven species evolution and morphological uniqueness within the family Goniodorididae.


Metabolomic investigations in cerebrospinal fluid of Parkinson's disease.
The underlying mechanisms of Parkinson´s disease are not completely revealed. Especially, early diagnostic biomarkers are lacking. To characterize early pathophysiological events, research is focusing on metabolomics. In this case-control study we investigated the metabolic profile of 31 Parkinson´s disease-patients in comparison to 95 neurologically healthy controls. The investigation of metabolites in CSF was performed by a 12 Tesla SolariX Fourier transform-ion cyclotron resonance-mass spectrometer (FT-ICR-MS). Multivariate statistical analysis sorted the most important biomarkers in relation to their ability to differentiate Parkinson versus control. The affected metabolites, their connection and their conversion pathways are described by means of network analysis. The metabolic profiling by FT-ICR-MS in CSF yielded in a good group separation, giving insights into the disease mechanisms. A total number of 243 metabolites showed an affected intensity in Parkinson´s disease, whereas 15 of these metabolites seem to be the main biological contributors. The network analysis showed a connection to the tricarboxylic cycle (TCA cycle) and therefore to mitochondrial dysfunction and increased oxidative stress within mitochondria. The metabolomic analysis of CSF in Parkinson´s disease showed an association to pathways which are involved in lipid/ fatty acid metabolism, energy metabolism, glutathione metabolism and mitochondrial dysfunction.


Pulmonary gas exchange is well preserved during robot assisted surgery in steep Trendelenburg position.
During robot assisted hysterectomies and prostatectomies, surgical exposure demands the application of a CO2 pneumoperitoneum with a very steep Trendelenburg position (40 degrees). The extent to which oxygenation and ventilation might be compromised intra-operatively remains poorly documented. Dead-space ventilation and venous admixture were determined in 18 patients undergoing robot assisted hysterectomy (n = 6) or prostatectomy (n = 12). Anesthesia was maintained with desflurane in O2 or O2/air, with the inspired O2 fraction left at the discretion of the attending anesthesiologist. Controlled mechanical ventilation was used, but 15 min after assuming the Trendelenburg position and up until resuming the supine position pressure controlled ventilation was used. Dead-space ventilation and venous admixture were determined using Bohr's formula and Nunn's iso-shunt diagram, respectively, at the following 7 stages of the procedure: 15 min after induction; 5 min after applying the CO2 pneumoperitoneum (intra-abdominal pressure 12 mm Hg) but while still supine; 5, 60, and 120 min after assuming the Trendelenburg positioning; and 5 and 15 min after reassuming the supine position. Venous admixture did not change. Dead-space ventilation increased after Trendelenburg positioning, and returned to baseline values after resuming the supine position. However, individual patterns varied widely. The lung has a remarkable yet incompletely understood capacity to withstand the effects of a CO2 pneumoperitoneum and steep Trendelenburg position during general anesthesia. While individual responses vary and should be monitored, effects on dead-space ventilation and venous admixture are small and should not be an obstacle to provide optimal surgical exposure during robot assisted prostatectomy or hysterectomy.


Nek6 mediates human cancer cell transformation and is a potential cancer therapeutic target.
We investigated the role of Nek6, a member of the NIMA-related serine/threonine kinase family, in tumorigenesis. Transcript, protein, and kinase activity levels of Nek6 were highly elevated in the malignant tumors and human cancer cell lines compared with normal tissue and fibroblast cells. Expression of exogenous wild-type Nek6 increased anchorage-independent growth of a variety of human cancer cell lines, whereas overexpression of the kinase-dead Nek6 and RNAi knockdown of endogenous Nek6 suppressed cancer cell transformation and induced apoptosis. Additionally, in in vivo xenograft nude mouse model, knockdown of Nek6 in HeLa cells resulted in reduction of tumor size relative to control siRNA tumors. Most importantly, knocking down endogenous Nek6 levels or exogenous expression of the kinase-dead form did not inhibit cell proliferation, nor did it induce apoptosis in normal fibroblast cells. Taken together, our data indicate a pivotal role for Nek6 in tumorigenesis and establish Nek6 as a potential target for treatment of a variety of human cancers.


Preparing the health workforce in Ethiopia: A Cross-sectional study of competence of anesthesia graduating students.
Efforts to address shortages of health workers in low-resource settings have focused on rapidly increasing the number of higher education programs for health workers. This study examines selected competencies achieved by graduating Bachelor of Science and nurse anesthetist students in Ethiopia, a country facing a critical shortage of anesthesia professionals. The study, conducted in June and July 2013, assessed skills and knowledge of 122 students graduating from anesthetist training programs at six public universities and colleges in Ethiopia; these students comprise 80% of graduates from these institutions in the 2013 academic year. Data was collected from direct observations of student performance, using an objective structured clinical examination approach, and from structured interviews regarding the adequacy of the learning environment. Student performance varied, with mean percentage scores highest for spinal anesthesia (80%), neonatal resuscitation (74%), endotracheal intubation (73%), and laryngeal mask airway insertion check (71%). Average scores were lowest for routine anesthesia machine check (37%) and preoperative screening assessment (48%). Male graduates outscored female graduates (63.2% versus 56.9%, P = 0.014), and university graduates outscored regional health science college graduates (64.5% versus 55.5%, P = 0.023). Multivariate linear regression found that competence was associated with being male and attending a university training program. Less than 10% of the students believed that skills labs had adequate staff and resources, and only 57.4% had performed at least 200 endotracheal intubations at clinical practicum sites, as required by national standards. Ethiopia has successfully expanded higher education for anesthetists, but a focus on quality of training and assessment of learners is required to ensure that graduates have mastered basic skills and are able to offer safe services.


Effect of bezafibrate on incidence of type 2 diabetes mellitus in obese patients.
To assess the effect of fibric acid derivative bezafibrate on incidence of type 2 diabetes in obese patients over a median 6.3 years follow-up period. The study sample comprised 339 non-diabetic obese patients (body mass index > or = 30.0 kg/m2) aged 42-74. Patients received either bezafibrate retard 400 mg (178 patients) or placebo (161 patients) once daily. Development of new diabetes was recorded in 98 patients: in 56 (37.0%) from the placebo group vs. 42 (27.1%) from the bezafibrate group, (P log-rank=0.01). The median time (interquartile range) until onset of new diabetes was significantly delayed in patients on bezafibrate when compared with those on placebo: 4.0 (2.1-5.0) vs. 2.0 (0.5-3.5) years, P=0.002. Multivariable analysis identified bezafibrate treatment as an independent predictor of reduced risk of new diabetes with hazard ratio (HR) 0.59 [95% confidence interval (CI) 0.39-0.91]. Other significant variables associated with future overt type 2 diabetes in obese patients were triglycerides (50 mg/dL increment) with HR 1.15 (95% CI 1.02-1.28) and fasting glucose (10 mg/dL increment) with HR 2.27 (95% CI 1.83-2.81). Bezafibrate, when compared with placebo, reduced the incidence and delayed the onset of type 2 diabetes in obese patients over a long-term follow-up period.


Sustained-release morphine sulfate with sequestered naltrexone for moderate to severe pain: a new opioid analgesic formulation and beyond.
Opioid usage during chronic nonmalignant pain has increased substantially over the past two decades. Prescription opioids have become the second most misused drug in the USA and prescription opioid abuse has escalated into a widespread public health problem. It is hoped that abuse-deterrent opioid formulations will take an important role in reducing opioid abuse, misuse and diversion. Embeda (sustained-release morphine sulfate with sequestered naltrexone)represents a new opioid formulation with an intended abuse-deterrent feature, now available on the market. Although Embeda seems to be a successful formulation by passing the efficacy trial, safety trial, pharmacokinetic study and abuse liability study, etc., it will require some long-term prospective epidemiological studies to substantiate fully its abuse-deterrent benefit. Embeda represents a new opioid formulation, adding to our arsenal to treat moderate to severe pain and playing its potential role in discouraging opioid abuse, misuse and diversion. Faced with an overwhelmingly expanding public health burden due to prescription opioid abuse, clinicians should always keep in mind the balance of maximizing pain relief and minimizing prescription opioid abuse.


Perinatal maternal mental health and infant socio-emotional development: A growth curve analysis using the MPEWS cohort.
Pregnancy and the early post partum period are widely understood as a critical period for the infant's emotional development and the earliest influence shaping social interaction. The present study aims to understand the potential influence of both antenatal and postnatal maternal anxiety and depressive symptoms on socio-emotional outcomes in offspring aged 12 months. The study used longitudinal data from a prospective cohort study on Australian pregnant women and their children. Data were available for 282 mothers and their children. Maternal depressive and anxiety symptoms were measured in early pregnancy, trimester three of pregnancy, six and 12 months postpartum. Social and emotional development in children was measured using the Brief Infant and Toddler Social Emotional Assessment (BITSEA) at 12 months. Using growth curve analysis of 4 waves of repeated measurement to examine intercept and slope, we found that both initial maternal depression and anxiety symptom levels, and the growth of these symptoms over time, predicted more problems with children's social and emotional development. In the final model anxiety accounted for 19% of the variance in child socio-emotional problems and depression 23% of variance. The results emphasise the importance of perinatal maternal mental health as a potential risk factor for child development. This carries important implications for policy development, such as the need to build early identification and early intervention models in to the current clinical practice for perinatal care, specifically, to develop targeted screening, assessment and interventions to address maternal mental health issues for at-risk parents during pregnancy, and continuing monitoring of young children whose mothers have experienced perinatal mental health difficulties.


Positive attitudes of farmers and pen-group conservation reduce adverse reactions of bulls during transfer for slaughter.
Transfer to slaughter may be very stressful for cattle and negatively affect their ultimate carcass pH. A potential link between farmers' attitudes, farmers' behaviour, responses of animals to handling and carcass pH in beef bull production was questioned. Whether carcass pH depends on experiences in handling and social mixing was analysed. We conducted a survey on commercial farms where we questioned farmers on their work and beliefs about bulls. Farmers' behaviour with bulls during a test and bulls' behaviour during loading in the truck for transport to the slaughterhouse were observed. The ultimate carcass pH was measured. Farmers tended to behave more gently with their bulls when they had positive attitudes towards gentle contacts with bulls (P = 0.07). The loading of bulls in the truck tended to be more difficult when the farmer was more ready to approach his bulls (P = 0.07). Carcass pH was higher for bulls that had not been transferred from a breeding to a finishing unit (P = 0.03). It tended to be higher when the farmer did not display a gentle behaviour (P = 0.09). The link between farmers' attitudes and farmers' behaviour and the lower meat pH resulting from a gentle farmers' behaviour during finishing are consistent with previous findings in pig, veal or dairy productions. However, the present links were weaker than in the other productions, probably due to the low frequency of close contacts between farmers and beef bulls. When loading bulls into a truck, handlers use the tendency of animals to avoid people, hence overly positive behaviour with the animals during finishing may result in more difficulties at loading. In that case, use of alternative driving aids should be recommended. Our results on carcass pH suggest some habituation to transport among bulls transferred between breeding and finishing, even though the two experiences are several months apart. In view of our results, it seems that contacts with bulls, when they are needed, should be gentle.


[Low birth weight, adrenal and sex hormones and their correlation with carbohydrate metabolism and cardiovascular physiology, investigated in young adulthood] .
It is known that the prevalence of cardiovascular diseases, hypertension, noninsulin dependent diabetes mellitus and dyslipidemia in the late adulthood are in connection with intrauterine retardation, characterized by low birth weight. One possible explanation of this phenomenon is the abnormality of hypothalamus-hypophysis-adrenal cortex axis due to the accelerated growth. The authors investigated the steroid levels of young adults; whom birth weight were under 2500 g, and examined the relationship between hormone levels and some parameters of glucose metabolism and cardiovascular system. 75 subjects (43 female and 32 male patients, mean age: 19.6 and 19.8 years, respectively; range 18-22 ys) with low birth weight and without any sign of chronic disease, and 30 healthy, age-matched controls with normal birth weight were investigated. The basal serum cortisol, dehydroepiandrosterone (DHEA), dehydroepiandrosterone sulphate (DHEAS), androstenedione (AD), 17-hydroxyprogesterone (17OHP), estradiol (OE), sex-hormone binding globulin (SHBG), FSH, LH and insulin levels were determined. Moreover, oral glucose tolerance test with 75 g glucose (OGTT), impedance cardiography as well as ambulatory blood pressure monitoring were done by all subjects. In both sexes in subjects with low birth weight the mean serum cortisol level was significantly higher, than in the normal controls. In female patients the serum DHEA, DHEAS, AD, and 17OHP levels were significantly higher than in the controls. Moreover, among these females a relationship was found between the elevations of adrenal and gonadal steroids and hyperinsulinemia, characterized by increased insulin response during OGTT. In male subjects a significant correlation was found between serum cortisol levels and systolic blood pressure and heart rate. In females there was a positive relationship between serum DHEA and heart rate. Summarized, the basic abnormality in patients with low birth weight seems to be a relative hypercortisolism, and in females because of hyperinsulinemia exists a mild hyperandrogenism as well. The hypercortisolism may cause cardiovascular abnormalities in males directly, while in females indirectly through the hyperinsulinemia and hyperandrogenism. These subtle abnormalities can be detected when no clinical signs present themselves, in young adulthood, giving the opportunity of taking preventive actions.


Host-parasite relationship of Ergasilus labracid Krøyer (Cyclopidea, Ergasilidae) and the striped bass, Morone saxatilis (Walbaum) from the lower Chesapeake Bay.
Epithelial hyperplasia was the most distinctive pathological change in gill filaments of striped bass, Morone saxatilis (Walbaum), from the lower Chesapeake Bay infested by Ergasilus labracis Krøyer. This tissue reaction initiated in the filament area most adjacent to the mouth of the attached parasite. Interruption of parasite egg sac production and eventual dislodgement associated with swelling of the gill filaments occurred unilaterally on gill arches of either side of the branchial basket. The largest number of parasites in all fish was found on the first outer hemibranch while abundance on successive hemibranches decreased in rank order. Copepods on the inner hemibranch, however, were least numerous on the first inner hemibranch and more numerous on the fourth inner hemibranch. On the whole, the number of parasites attached to the outer hemibranch outnumbered the number of parasites established on the inner hemibranchs. This differential settlement was most evident in low to moderate infections and became less apparent in heavy infections. Newly settled copepods showed highest abundance on the third arch and least abundance on the first and fourth arches. The causes for this differential distribution on the gill arches are discussed.


Mantle cell lymphoma salvage regimen: synergy between a reprogrammed oncolytic virus and two chemotherapeutics.
Measles virus (MV)-PNP H(blind)antiCD20 is a CD20-targeted and prodrug convertase-armed MV that temporarily controls growth of lymphoma xenografts in severe combined immunodeficiency (SCID) mice in combination with fludarabine phosphate (fludarabine). Herein, we examine the replication of this targeted virus and of a vaccine-lineage MV in disease bulks and circulating cells from mantle cell lymphoma (MCL) patients, and show that only the targeted virus is specific for CD20-expressing cells. We then assessed the efficacy of different regimens of administration of this virus in combination with fludarabine and cyclophosphamide (CPA) in an MCL xenograft model. We show that CPA administration before the beginning of virus treatment enhances oncolytic efficacy, likely through temporary immunosuppression. An interval of 1 week between intravenous virus administration and fludarabine treatment further enhanced oncolysis, by synchronizing maximum prodrug convertase expression with fludarabine availability. Finally, three 23-day courses of triple sequential treatment with CPA, virus and fludarabine treatment resulted in complete regression of the xenografts. Secondary disease symptoms interfered with survival, but average survival times increased from 22 to 77 days. These studies document a reprogrammed oncolytic virus, consolidating the effects of two chemotherapeutics, a concept well suited for a phase I clinical trial for MCL patients for whom conventional therapies have failed.


Burn scar contracture release surgery effectively improves functional range of motion, disability and quality of life: A pre/post cohort study with long-term follow-up in a Low- and Middle-Income Country.
Burn scar contractures limit range of motion (ROM) of joints and have substantial impact on disability and the quality of life (QoL) of patients, particularly in a Low- and Middle-Income Country (LMIC) setting. Studies on the long-term outcome are lacking globally; this study describes the long-term impact of contracture release surgery performed in an LMIC. This is a pre-post cohort study, conducted in a referral hospital in Tanzania. Patients who underwent burn scar contracture release surgery in 2017-2018 were eligible. ROM (goniometry), disability (WHODAS 2.0) and QoL (EQ-5D) were assessed. The ROM data were compared to the ROM that is required to perform activities of daily living without compensation, i.e. functional ROM. Assessments were performed preoperatively and at 1, 3, 6 and 12 months postoperatively. In total, 44 patients underwent surgery on 115 affected joints. At 12 months, the follow-up rate was 86%. The mean preoperative ROM was 37.3% of functional ROM (SD 31.2). This improved up to 108.7% at 12 months postoperatively (SD 42.0, p < 0.001). Disability-free survival improved from 55% preoperatively to 97% at 12 months (p < 0.001) postoperatively. QoL improved from 0.69 preoperatively, to 0.93 (max 1.0) at 12 months postoperatively (p < 0.001). Patients who regained functional ROM in all affected joints reported significantly less disability (p < 0.001) and higher QoL (p < 0.001) compared to patients without functional ROM. Contracture release surgery performed in an LMIC significantly improved functional ROM, disability and QoL. Results showed that regaining a functional joint is associated with less disability and higher QoL.


Biochemical Features of Beneficial Microbes: Foundations for Therapeutic Microbiology.
Commensal and beneficial microbes secrete myriad products which target the mammalian host and other microbes. These secreted substances aid in bacterial niche development, and select compounds beneficially modulate the host and promote health. Microbes produce unique compounds which can serve as signaling factors to the host, such as biogenic amine neuromodulators, or quorum-sensing molecules to facilitate inter-bacterial communication. Bacterial metabolites can also participate in functional enhancement of host metabolic capabilities, immunoregulation, and improvement of intestinal barrier function. Secreted products such as lactic acid, hydrogen peroxide, bacteriocins, and bacteriocin-like substances can also target the microbiome. Microbes differ greatly in their metabolic potential and subsequent host effects. As a result, knowledge about microbial metabolites will facilitate selection of next-generation probiotics and therapeutic compounds derived from the mammalian microbiome. In this article we describe prominent examples of microbial metabolites and their effects on microbial communities and the mammalian host.


Cost and effectiveness of Chlamydia screening among male military recruits: Markov modeling of complications averted through notification of prior female partners.
Despite rising rates of female screening, a high economic burden remains associated with Chlamydia infection from high rates of undetected asymptomatic disease and its associated sequelae of pelvic inflammatory disease (PID) and chronic pelvic pain (CP). Males comprise the majority of US military recruits and represent an ideal population in which to achieve identification and interruption of sexually transmitted infection among infected partners through mass tandem screening. We developed a static decision tree incorporating a calibrated Markov model to predict the differences in healthcare payer direct healthcare costs, cases of PID and CP averted among female partners of male recruits through implementation of either selective (aged 24 and younger) or universal recruit screening policies incorporating partner notification. A policy of selective male screening added $10.30 in direct costs per recruit, whereas universal male screening added an additional $1.60. A policy of selective male screening yielded an incremental cost-effectiveness ratio of $3.7K per case of PID averted, and $7.3K per case of CP averted, whereas universal screening yielded an incremental cost-effectiveness ratio of $8.2K per additional case of PID and $16.4K per additional case of CP averted beyond selective screening. Neither policy was dominant, and results were qualitatively robust to single-variable and probabilistic sensitivity analysis. In consonance with other studies of mass tandem screening, we found both selective and universal male recruit screening cost-effective as compared with other interventions. Our results argue in favor of universal screening of male recruits for Chlamydia infection, linked to partner notification.


Diagnosis and treatment of minimal brain dysfunction in adults: a preliminary report.
Minimal brain dysfunction (MBD) has long been considered a disorder limited to childhood. A number of longitudinal and adoption studies suggest that MBD may persist into adult life where its existence is concealed by the application of a variety of diagnostic labels. In order to test the hypothesis that MBD does persist into adulthood, 15 putative MBD adults were identified on the basis of current MBD-like complaints, self-description of MBD characteristics in childhood, and a parental rating (on a standardized form) of "hyperactivity" in childhood. Eleven of the 15 subjects were given a double-blind trial of methylphenidate hydrochloride, and all 15 were given an open trial of pemoline, imipramine hydrochloride, or amitryptiline hydrochloride. Eight of the 11 showed a significant response to the double-blind trial of methylphenidate. Of the 15, eight showed a good response to stimulants or tricyclic antidepressants, two showed a moderately favorable response, and five were unresponsive to drug therapy.


Drug Interactions at the Human Placenta: What is the Evidence?
Pregnant women (and their fetuses) are treated with a significant number of prescription and non-prescription medications. Interactions among those drugs may affect their efficacy and toxicity in both mother and fetus. Whereas interactions that result in altered drug concentrations in maternal plasma are detectable, those involving modulation of placental transfer mechanisms are rarely reflected by altered drug concentrations in maternal plasma. Therefore, they are often overlooked. Placental-mediated interactions are possible because the placenta is not only a passive diffusional barrier, but also expresses a variety of influx and efflux transporters and drug-metabolizing enzymes. Current data on placental-mediated drug interactions are limited. In rodents, pharmacological or genetic manipulations of placental transporters significantly affect fetal drug exposure. In contrast, studies in human placentae suggest that the magnitude of such interactions is modest in most cases. Nevertheless, under certain circumstances, such interactions may be of clinical significance. This review describes currently known mechanisms of placental-mediated drug interactions and the potential implications of such interactions in humans. Better understanding of those mechanisms is important for minimizing fetal toxicity from drugs while improving their efficacy when directed to treat the fetus.


Cytosolic processing of proteasomal cleavage products can enhance the presentation efficiency of MHC-1 epitopes.
The vertebrate immune system is able to detect abnormal body cells by the specific repertoire of 8 - 12 residues long peptides (= epitopes or peptide antigens) presented at the cell surface by the MHC-1 molecule complex. The generation of an epitope starts with the degradation of endogenous proteins into primary oligomeric fragments by cytosolic proteases, predominantly the proteasome. These primary fragments may be further attacked by various amino peptidases resident in the cytosol or, alternatively, may escape from this attack by entering the endoplasmic reticulum (ER) by the transporter associated with antigen presentation (TAP). To study the possible consequences of this scenario for the efficiency of antigen presentation we have applied kinetic modelling. The mathematical model comprises the generation of primary oligomeric fragments containing the definitive epitope, the successive N-terminal shortening of these primary fragments by cytosolic amino peptidases and the TAP-mediated transport of cytosolic peptides into the ER. Because the number of peptide molecules may become very small we have performed deterministic and stochastic simulations of the kinetic model. Our simulations show that cytosolic N-terminal trimming of primary fragments may drastically increase loading epitope precursors into the ER. In particular, a primary fragment generated with a low rate of TAP transport into the ER may nevertheless become a potent epitope precursor if at least one of its N-terminal trimming products will be efficiently transported.


Kinetic analysis of the thermal processing of silica and organosilica.
The incorporation of an organic group into sol-gel-derived silica causes significant changes in the structure and properties of these materials. Therefore, the thermal treatment of organosilica materials may require a different approach. In the present paper, kinetic parameters (activation energy, pre-exponential constant, and reaction models) have been determined from mass loss data for the dehydration, dehydroxylation, and decomposition reactions that take place upon heating silica and organosilica. Parameters were obtained by employing model-free isoconversional methods to data obtained under multiple heating rates as well as by multivariate analysis of the kinetics using a multistep reaction model with distributed activation energy. For silica, it can be concluded that the reaction atmosphere (i.e., inert or thermo-oxidative) has no influence on the reaction rate of the dehydration and dehydroxylation reactions that are responsible for the densification of the material. Under inert atmosphere, full dehydration can be reached without affecting the organic moiety. Achieving complete dehydroxylation of the organosilica is practically impossible as decomposition does manifest itself under commonly employed calcination temperatures. This indicates that prudence is required in designing a heat treatment program for these hybrid materials. To aid in optimizing the thermal treatment, a predictive model was developed, which can be used to forecast the extent of dehydration, dehydroxylation, and decomposition reactions under a multitude of temperature programs.


siRNA targeting the leader sequence of SARS-CoV inhibits virus replication.
SARS-CoV (the SARS-Associated Coronavirus) was reported as a novel virus member in the coronavirus family, which was the cause of severe acute respiratory syndrome. Coronavirus replication occurs through a unique mechanism employing Leader sequence in the transcripts when initiating transcription from the genome. Therefore, we cloned the Leader sequence from SARS-CoV(BJ01), which is identical to that identified from SARS-CoV(HKU-39849), and constructed specific siRNA targeting the Leader sequence. Using EGFP and RFP reporter genes fused with the cloned SARS-CoV Leader sequence, we demonstrated that the siRNA targeting the Leader sequence decreased the mRNA abundance and protein expression levels of the reporter genes in 293T cells. By stably expressing the siRNA in Vero E6 cells, we provided data that the siRNA could effectively and specifically decrease the mRNA abundance of SARS-CoV genes as analyzed by RT-PCR and Northern blot. Our data indicated that the siRNA targeting the Leader sequence inhibited the replication of SARS-CoV in Vero E6 cells by silencing gene expression. We further demonstrated, via transient transfection experiments, that the siRNA targeting the Leader sequence had a much stronger inhibitory effect on SARS-CoV replication than the siRNAs targeting the Spike gene or the antisense oligodeoxynucleotides did. This report provides evidence that targeting Leader sequence using siRNA could be a powerful tool in inhibiting SARS-CoV replication.


iFoldRNA v2: folding RNA with constraints.
A key to understanding RNA function is to uncover its complex 3D structure. Experimental methods used for determining RNA 3D structures are technologically challenging and laborious, which makes the development of computational prediction methods of substantial interest. Previously, we developed the iFoldRNA server that allows accurate prediction of short (<50 nt) tertiary RNA structures starting from primary sequences. Here, we present a new version of the iFoldRNA server that permits the prediction of tertiary structure of RNAs as long as a few hundred nucleotides. This substantial increase in the server capacity is achieved by utilization of experimental information such as base-pairing and hydroxyl-radical probing. We demonstrate a significant benefit provided by integration of experimental data and computational methods. http://ifoldrna.dokhlab.org email@example.com.


Vitamin D receptor genotype is associated with cortical bone loss in Japanese patients with primary hyperparathyroidism.
Vitamin D receptor (VDR) genotype was determined in 66 Japanese patients with primary hyperparathyroidism (pHPT). In contrast to previous report showing an association between VDR genotype bb and the development of pHPT in a Swedish population, we did not find any differences in the frequencies of the VDR genotypes between pHPT and the control. The bone mineral density in the radius was significantly lower (Z score, -1.723 +/- 1.819) in bb genotype than in BB/Bb genotype (+0.255 +/- 2.344) in pHPT patients. Based on the fact that PTH requires vitamin D3 to take effect on calcium mobilization from bone, it is possible that VDR polymorphism may influence the PTH action on bone.


Effects of a Saccharomyces cerevisiae fermentation product on fecal characteristics, metabolite concentrations, and microbiota populations of dogs subjected to exercise challenge.
The objective of this study was to determine the fecal characteristics, microbiota, and metabolites of dogs fed a Saccharomyces cerevisiae fermentation product (SCFP) and subjected to exercise challenge in untrained and trained states. Thirty-six adult dogs (18 male, 18 female; mean age: 7.1 yr; mean body weight: 29.0 kg) were randomly assigned to control or SCFP-supplemented (250 mg/dog/d) diets and fed for 10 wk. After 3 wk, dogs were given an exercise challenge (6.5 km run), with fresh fecal samples collected pre- and post-challenge. Dogs were then trained by a series of distance-defined running exercise regimens over 7 wk (two 6.4 km runs/wk for 2 wk; two 9.7 km runs/wk for 2 wk; two 12.9 km runs/wk for 2 wk; two 3.2 km runs/wk). Dogs were then given exercise challenge (16 km run) in the trained state, with fresh fecal samples collected pre- and post-challenge. Fecal microbiota data were evaluated using QIIME2, while all other data were analyzed using the Mixed Models procedure of SAS. Effects of diet, exercise, and diet*exercise were tested with P < 0.05 considered significant. Exercise challenge reduced fecal pH and ammonia in both treatments, and in untrained and trained dogs. After the exercise challenge in untrained dogs, fecal indole, isobutyrate, and isovalerate were reduced, while acetate and propionate were increased. Following the exercise challenge in trained dogs, fecal scores and butyrate decreased, while isobutyrate and isovalerate increased. SCFP did not affect fecal scores, pH, dry matter, or metabolites, but fecal Clostridium was higher in controls than in SCFP-fed dogs over time. SCFP and exercise challenge had no effect on alpha or beta diversity in untrained dogs. However, the weighted principal coordinate analysis plot revealed clustering of dogs before and after exercise in trained dogs. After exercise challenge, fecal Collinsella, Slackia, Blautia, Ruminococcus, and Catenibacterium were higher and Bacteroides, Parabacteroides, Prevotella, Phascolarctobacterium, Fusobacterium, and Sutterella were lower in both untrained and trained dogs. Using qPCR, SCFP increased fecal Turicibacter, and tended to increase fecal Lactobacillus vs. controls. Exercise challenge increased fecal Turicibacter and Blautia in both untrained and trained dogs. Our findings show that exercise and SCFP may affect the fecal microbiota of dogs. Exercise was the primary cause of the shifts, however, with trained dogs having more profound changes than untrained dogs.