The interface of teaching, research and extension in undergraduate courses in health.
This article aimed to describe and evaluate the extension program "Integrating knowledge to the family integral care" composed of five projects that aimed to enrich knowledge and develop skills in the academic field, to accomplish health actions, based on the families' and community' needs. A total of 33 faculties from the nursing, medical and biomedical schools at the "Triângulo Mineiro Federal University" participated in the projects during eight months in the "Family Health Program", out-patient clinics and at the School Hospital, attending 2,000 clients. The faculty members were subjects in their own learning process, experiencing and reflecting upon this social reality. The program also favored inter-institutional integration, as well as, showed connection between teaching-research-extension.


Postdiagnosis Loss of Skeletal Muscle, but Not Adipose Tissue, Is Associated with Shorter Survival of Patients with Advanced Pancreatic Cancer.
Pancreatic cancer is associated with development of cachexia, a wasting syndrome thought to limit survival. Few studies have longitudinally quantified peripheral tissues or identified biomarkers predictive of future tissue wasting. Adipose and muscle tissue were measured by computed tomography (CT) at diagnosis and 50 to 120 days later in 164 patients with advanced pancreatic cancer. Tissue changes and survival were evaluated by Cox proportional hazards regression. Baseline levels of circulating markers were examined in relation to future tissue wasting. Compared with patients in the bottom quartile of muscle change per 30 days (average gain of 0.8 ± 2.0 cm2), those in the top quartile (average loss of 12.9 ± 4.9 cm2) had a hazard ratio (HR) for death of 2.01 [95% confidence interval (CI), 1.12-3.62]. Patients in the top quartile of muscle attenuation change (average decrease of 4.9 ± 2.4 Hounsfield units) had an HR of 2.19 (95% CI, 1.18-4.04) compared with those in the bottom quartile (average increase of 2.4 ± 1.6 Hounsfield units). Changes in adipose tissue were not associated with survival. Higher plasma branched chain amino acids (BCAA; P = 0.004) and lower monocyte chemoattractant protein-1 (MCP-1; P = 0.005) at diagnosis were associated with greater future muscle loss. In patients with advanced pancreatic cancer, muscle loss and decrease in muscle density in 2 to 4 months after diagnosis were associated with reduced survival. BCAAs and MCP-1 levels at diagnosis were associated with subsequent muscle loss. BCAAs and MCP-1 levels at diagnosis could identify a high-risk group for future tissue wasting.


Reinke's edema: investigations on the role of MIB-1 and hepatocyte growth factor.
Reinke's edema is a benign disease of the human vocal fold, which mainly affects the sub-epithelial layer of the vocal fold. Microscopic observations show a strongly oedematous epithelium with loosened intercellular junctions, a disruption of the extracellular connections between mucosal epithelium and connective tissue, closely adherent to the thyroarytenoid muscle. Thickening of the basal layer of epithelium, known as Reinke's space, high deposition of fibronectin and chronic inflammatory infiltration it is also visible. We analyzed, together with the hepatocyte growth factor (HGF), the expression level of MIB-1 in samples harvested from patients affected by Reinke's edema, in order to define its biological role and consider it as a possible prognostic factor in the follow-up after surgical treatment. We observed a moderate expression of HGF in the lamina propria of the human vocal fold and in the basal membrane of the mucosal epithelium. Our finding suggests that this growth factor acts as an antifibrotic agent in Reinke's space and affects the fibronectin deposition in the lamina propria. MIB-1, on the contrary, showed a weak expression in the basement membrane of the mucosal epithelium and a total absence in the lamina propria deep layer, thus suggesting that only the superficial layer is actively involved in the reparatory process with a high regenerative capacity, together with a high deposition of fibronectin. The latter is necessary for the cellular connections reconstruction, after the inflammatory infiltration.


Renal function in the elderly.
Renal mass, glomerular filtration rate and renal blood flow decline with age. Since the renal tubules also decrease in mass, the aging kidney is less able to compensate for changes in fluid balance. Because of changes in the renin-aldosterone system, the elderly may have problems maintaining sodium and potassium balance. Physicians must be aware of the implications of these changes in renal function when they prescribe medications or order intravenous fluids for elderly patients.


Treatment of Class II malocclusion with 2-maxillary premolar extractions: Monthly vs biweekly appointments, which is more efficient?
This study compared whether there is a difference in treatment time (TT) and efficiency when appointments are held once a month or at 2-week intervals in patients with Class II malocclusion treated with 2-premolar extractions METHODS: The patients of this retrospective sample were treated with the same orthodontic mechanics and divided into 2 groups according to frequencies of orthodontic appointments. Group 1 consisted of 18 patients (10 males, 8 females), with an initial mean age of 14.38 ± 1.38 years and appointments once a month. Group 2 consisted of 19 patients (9 males, 10 females), with an initial mean age of 14.12 ± 1.38 years and biweekly appointments. Edgewise fixed appliances with a slot size of 0.022 × 0.028 mm were used, and the anterior teeth were retracted en-masse with a rectangular wire and elastic chains. The Peer Assessment Rating and Objective Grading System indexes were measured in the dental models at the beginning and the end of treatment. Efficiency was assessed by dividing the percentage of improvement of each occlusal index concerning the multiplication of TT and the number of appointments. Both groups presented similar Peer Assessment Rating and Objective Grading System indexes at the end of treatment. Groups 1 and 2 presented different TTs (28.06 and 22.05 months, respectively); however, there were no differences regarding the efficiency of both protocols. Patients with Class II malocclusion treated with 2-maxillary premolars had significantly shorter treatment times when seen on biweekly appointments than those with monthly appointments; however, there was no difference in efficiency between protocols.


Bcl-Xshort is elevated following severe global ischemia in rat brains.
Hippocampal CA1 neurons are highly susceptible to short periods of transient global ischemia. We have previously reported in a rat model of transient forebrain global ischemia that activation and nuclear localization of NF-kB occurs in the CA1 neurons at 24 and 72 h post reperfusion. Events following NF-kB activation would ultimately determine whether damaged cells will undergo programmed cell death. We have selected bcl-x gene expression for study because there is increasing evidence that proteins encoded by the bcl-2 gene family (bcl-2, bcl-x, bax etc) play a role in the regulation of programmed cell death. We have observed that the bcl-x gene promoter contains a putative consensus sequence for NF-kB/CS4 responsive activation. We also can show that other members of the bcl-2 multigene family contain the NF-kB/CS4 sequence in their five prime regulatory regions. In this study, we show that NF-kB p50 and NF-kB p65 act in synergy to transactivate the bcl-x promoter in co-transfected 293 cells. We also report that following ischemia and NF-kB activation, bcl-x messenger RNA levels increase in the CA1 hippocampal region. As a result of this transcriptional increase, surprisingly, it is bcl-xs, the apoptotic form of bcl-x, that is elevated. These results suggest that activation of NF-kB can lead to increased expression of bcl-x as manifested by the increase in the short form of bcl-x.


The functional DRD3 Ser9Gly polymorphism (rs6280) is pleiotropic, affecting reward as well as movement.
Abnormalities of motivation and behavior in the context of reward are a fundamental component of addiction and mood disorders. Here we test the effect of a functional missense mutation in the dopamine 3 receptor (DRD3) gene (ser9gly, rs6280) on reward-associated dopamine (DA) release in the striatum. Twenty-six healthy controls (HCs) and 10 unmedicated subjects with major depressive disorder (MDD) completed two positron emission tomography (PET) scans with [(11)C]raclopride using the bolus plus constant infusion method. On one occasion subjects completed a sensorimotor task (control condition) and on another occasion subjects completed a gambling task (reward condition). A linear regression analysis controlling for age, sex, diagnosis, and self-reported anhedonia indicated that during receipt of unpredictable monetary reward the glycine allele was associated with a greater reduction in D2/3 receptor binding (i.e., increased reward-related DA release) in the middle (anterior) caudate (p<0.01) and the ventral striatum (p<0.05). The possible functional effect of the ser9gly polymorphism on DA release is consistent with previous work demonstrating that the glycine allele yields D3 autoreceptors that have a higher affinity for DA and display more robust intracellular signaling. Preclinical evidence indicates that chronic stress and aversive stimulation induce activation of the DA system, raising the possibility that the glycine allele, by virtue of its facilitatory effect on striatal DA release, increases susceptibility to hyperdopaminergic responses that have previously been associated with stress, addiction, and psychosis.


An observational study of the success and complications of 2546 external cephalic versions in low-risk pregnant women performed by trained midwives.
To evaluate the success of an external cephalic version (ECV) training programme, and to determine the rates of successful ECV, complications, and caesarean birth in a low-risk population. Prospective observational study. Primary health care and hospital settings throughout the Netherlands (January 2008-September 2011). Low-risk women with a singleton fetus in breech presentation, without contraindications to ECV, were offered ECV at approximately 36 weeks of gestation. Data were collected for all ECVs performed by midwives, and were entered into a national online database. Successful ECV was defined as the fetus having a cephalic presentation immediately following the procedure and at birth. Complications were observed at ≤ 30 minutes and between 30 minutes and 48 hours after the ECV procedure. All serious pregnancy outcomes that occurred after the ECV procedure until birth were reported. A total of 47% had a successful ECv and a cephalic at the time of birth: 34% of nulliparous and 66% of multiparous women. After ECV, 57% of women gave birth vaginally: 45% of nulliparous women and 76% of multiparous women. Within 30 minutes after ECV, and between 30 minutes and 48 hours after ECV, the proportion of women experiencing a complication or serious pregnancy outcome was 0.9% and 1.8%, respectively. Serious pregnancy outcome at any time following ECV until birth was experienced by 58 (2.5%) of the women. The success rate of ECVs performed by trained midwives in primary health care or hospital settings is comparable with that of other providers, and the procedure is safe for low-risk women.


How did the swiss cheese plant get its holes?
Adult leaf fenestration in "Swiss cheese" plants (Monstera Adans.) is an unusual leaf shape trait lacking a convincing evolutionary explanation. Monstera are secondary hemiepiphytes that inhabit the understory of tropical rainforests, where photosynthesis from sunflecks often makes up a large proportion of daily carbon assimilation. Here I present a simple model of leaf-level photosynthesis and whole-plant canopy dynamics in a stochastic light environment. The model demonstrates that leaf fenestration can reduce the variance in plant growth and thereby increase geometric mean fitness. This growth-variance hypothesis also suggests explanations for conspicuous ontogenetic changes in leaf morphology (heteroblasty) in Monstera, as well as the absence of leaf fenestration in co-occurring juvenile tree species. The model provides a testable hypothesis of the adaptive significance of a unique leaf shape and illustrates how variance in growth rate could be an important factor shaping plant morphology and physiology.


Correlation of sister chromatid exchange frequency with mutation induction and cell survival in Chinese hamster ovary cells exposed to N-oxidized arylamines.
In this study the relationships between sister chromatid exchange (SCE) frequency, mutation induction at the hypoxanthineguanine phosphoribosyl transferase locus, and cell survival were established in Chinese hamster ovary cells exposed to one of the three N-oxidized arylamines. The toxicants used were N-hydroxy-2-aminofluorene, N-hydroxy-N'-acetylbenzidine, and 1-nitrosopyrene. Mutation induction as measured by resistance to 6-thioguanine related poorly to reduced cloning ability and SCE frequency. Although SCEs were formed at concentrations of toxicants which produced no measurable reduction in cell survival, a strong correlation was observed between these two biological responses. This relationship was strengthened when the results obtained in this study were combined with those from a previous study which examined the relationship between SCE induction and cell survival in Chinese hamster ovary cells exposed to simple alkylating agents. These results support the contention that a common step is involved in the induction of SCE and cellular toxicity in Chinese hamster ovary cells exposed to certain classes of chemical toxicants.


Cathepsin K analysis in a pycnodysostosis cohort: demographic, genotypic and phenotypic features.
To characterize cathepsin K (CTSK) mutations in a group of patients with pycnodysostosis, who presented with either short stature or atypical fractures to pediatric endocrinology or dysmorphic features to pediatric genetics clinics. Seven exons and exon/intron boundaries of CTSK gene for the children and their families were amplified with PCR and sequenced. Sixteen patients from 14 families with pycnodysostosis, presenting with typical dysmorphic features, short stature, frequent fractures and osteosclerosis, were included in the study. We identified five missense mutations (M1I, I249T, L7P, D80Y and D169N), one nonsense mutation (R312X) and one 301 bp insertion in intron 7, which is revealed as Alu sequence; among them, only L7P and I249 were described previously. The mutations were homozygous in all cases, and the families mostly originated from the region where consanguineous marriage rate is the highest. Patients with M1I mutation had fractures, at younger ages than the other pycnodysostosis cases in our cohort which were most probably related to the severity of mutation, since M1I initiates the translation, and mutation might lead to the complete absence of the protein. The typical finding of pycnodysostosis, acroosteolysis, could not be detected in two patients, although other patients carrying the same mutations had acroosteolysis. Additionally, none of the previously described hot spot mutations were seen in our cohort; indeed, L7P and R312X were the most frequently detected mutations. We described a large cohort of pycnodysostosis patients with genetic and phenotypic features, and, first Alu sequence insertion in pycnodysostosis.


Building a Culture of Intimate Partner Violence Prevention in Alberta, Canada Through the Promotion of Healthy Youth Relationships.
This article explores the design and implementation of the Alberta Healthy Youth Relationships (AHYR) Strategy. The AHYR is a province-wide practice and policy change initiative in Alberta, Canada, that aims to prevent intimate partner violence by promoting service provider and systems capacity to support healthy relationships in adolescence and beyond. Developed in 2012 within a broader initiative called Shift: The Project to End Domestic Violence, the design of the AHYR began with work with provincial policy-makers, in order to create a policy climate that championed primary prevention practice. This policy climate subsequently supported the province-wide implementation of three evidence-based/evidence-informed programs that focus on building the skills and capacities required for healthy youth relationships. Through these programs, the AHYR has reached over 62,000 youth in grades 7-9, 900 teachers, 850 parents, and 1300 adults that work with youth across the province. In addition to these three programs, the AHYR also works with larger systems (e.g., policymakers, local funders, post-secondary institutions) to advance primary prevention practice. In this paper, we describe how the AHYR contributed to a culture of intimate partner violence prevention practice in the province by improving the readiness of funders and system leaders, community organizations, and practitioners to support healthy relationships best practices. We also describe how we used process evaluation to explore the potential for practice change and to inform the design of the next iteration of the AHYR. The article concludes with implications for other researchers and practitioners aiming to build a culture of intimate partner violence prevention practice within their province or state.


Role of nox2-based NADPH oxidase in bone marrow and progenitor cell function involved in neovascularization induced by hindlimb ischemia.
Bone marrow (BM) is the major reservoir for endothelial progenitor cells (EPCs). Postnatal neovascularization depends on not only angiogenesis but also vasculogenesis, which is mediated through mobilization of EPCs from BM and their recruitment to the ischemic sites. Reactive oxygen species (ROS) derived from Nox2-based NADPH oxidase play an important role in postnatal neovascularization; however, their role in BM and EPC function is unknown. Here we show that hindlimb ischemia of mice significantly increases Nox2 expression and ROS production in BM-mononuclear cells (BMCs), which is associated with an increase in circulating EPC-like cells. Mice lacking Nox2 show reduction of ischemia-induced flow recovery, ROS levels in BMCs, as well as EPC mobilization from BM. Transplantation of wild-type (WT)-BM into Nox2-deficient mice rescues the defective neovascularization, whereas WT mice transplanted with Nox2-deficient BM show reduced flow recovery and capillary density compared to WT-BM transplanted control. Intravenous infusion of WT- and Nox2-deficient BMCs into WT mice reveals that neovascularization and homing capacity are impaired in Nox2-deficient BMCs in vivo. In vitro, Nox2-deficient c-kit+Lin- BM stem/progenitor cells show impaired chemotaxis and invasion as well as polarization of actins in response to stromal derived factor (SDF), which is associated with blunted SDF-1-mediated phosphorylation of Akt. In conclusion, Nox2-derived ROS in BM play a critical role in mobilization, homing, and angiogenic capacity of EPCs and BM stem/progenitor cells, thereby promoting revascularization of ischemic tissue. Thus, NADPH oxidase in BM and EPCs is potential therapeutic targets for promoting neovascularization in ischemic cardiovascular diseases.


Quantifying ruminal digestion of organic matter and neutral detergent fiber using the omasal sampling technique in cattle--a meta-analysis.
A data set from 32 studies (122 diets) was used to evaluate the accuracy and precision of the omasal sampling technique by investigating the relationships between ruminal and total digestion of neutral detergent fiber (NDF), between intake and apparent and true ruminal digestion of organic matter (OM), and between omasal NAN flow and milk protein yield. A mixed model regression analysis with random study effect was used to evaluate the relationships. The data were obtained when feeding North American diets (n=36) based on alfalfa silage, corn silage, and corn grain and North European diets (n=86) comprising grass silage supplemented with barley-based concentrates. In all studies, digesta flow was quantified using a triple-marker approach. Standard deviations of ruminal NDF and true OM digestibility were smaller than typically reported in duodenal sampling studies using only chromic oxide as a flow marker. The relationship between total and ruminal NDF digestion was consistent, indicating little variation in the proportion of total-tract NDF digestion that occurred in the rumen. Furthermore, the slope of this regression indicated that 94.7% (+/-2.7%) of total NDF digestion occurred in the rumen. The slopes of mixed model regression equations between OM intake and amount digested indicated that 42% (+/-2.4%) and 74% (+/-3.1%) of OM was apparently and truly digested in the rumen, respectively. The contribution of the rumen to total-tract apparent OM digestion was 62% (+/-2.6%). The close relationship between omasal flow of nonammonia crude protein and milk protein yield (with adjusted residual mean squared error=31 g) provided further confidence in the reliability of omasal flow measurements.


Theoretical discovery of novel two-dimensional VA-N binary compounds with auxiticity.
Auxetic materials, which possess a negative Poisson's ratio (NPR), have been a hot topic in materials science research. Through atomistic simulations, we theoretically rediscover a few novel two-dimensional (2D) VA-nitride (VA-N) binary compounds with δ-phosphorene-like structures. The structures in the δ-phase (except for δ-PN) exhibit better stability in terms of energy, thermodynamics, and mechanics with respect to their counterparts in the α- and β-phases. The structures in the δ-phase show semiconducting behaviors with direct band gaps falling in the visible light region. Interestingly, most structures in the α- and δ-phases (except for δ-BiN) exhibit large in-plane NPRs and excellent mechanical properties. The maximum NPR occurs along the zigzag (x) direction for the δ-phases and along the diagonal direction for the α-phases. Particularly, for α- and δ-SbN, the NPRs are -0.628 and -0.296, respectively. δ-SbN can sustain tensile strains of up to 22% and 35% with maximum stresses of 12.1 and 9.8 GPa in the zigzag and armchair directions, respectively. In addition, the transverse response can reach up to 6.6% at a strain of ∼18% along the armchair (y) direction for δ-SbN, which is considerably higher than those of other 2D auxetic materials. Our results reveal that 2D VA-N binary compounds have potential applications in designing 2D electromechanical and optoelectronic devices.


New twist on the regulation of NKG2D ligand expression.
The NK cell-activating receptor NKG2D plays a prominent role in antitumor immune responses. Expression of the multiple NKG2D ligands must be tightly controlled to guarantee that NK cells attack tumors but not healthy cells. New data reveal a novel mechanism of posttranslational regulation of the mouse NKG2D ligand MULT1, in which MULT1 is ubiquitinated and degraded in healthy cells. In response to UV stress or heat shock, ubiquitination of MULT1 decreases and cell surface expression increases. Thus, targeting the ubiquitination machinery in cancer cells might increase the susceptibility of tumors to NK cell-mediated killing.


Transient Dynamics of d-Wave Superconductors after a Sudden Excitation.
Motivated by recent ultrafast pump-probe experiments on high-temperature superconductors, we discuss the transient dynamics of a d-wave BCS model after a quantum quench of the interaction parameter. We find that the existence of gap nodes, with the associated nodal quasiparticles, introduces a decay channel which makes the dynamics much faster than in the conventional s-wave model. For every value of the quench parameter, the superconducting gap rapidly converges to a stationary value smaller than the one at equilibrium. Using a sudden approximation for the gap dynamics, we find an analytical expression for the reduction of spectral weight close to the nodes, which is in qualitative agreement with recent experiments.


Spontaneous radiographic resolution and subsequent redemonstration of an untreated glioblastoma.
The authors present the first case of spontaneous radiographic resolution and subsequent redemonstration of Gd enhancement of an untreated glioblastoma. They also review the literature on MR imaging enhancement patterns of high-grade gliomas and speculate that this phenomenon could be attributed to the dynamic biology of glioblastomas but could also represent pseudoregression following successful control of seizure activity. A 57-year-old woman presented with left-sided paresthesias and numbness. Initial Gd-enhanced MR images exhibited T2 and FLAIR signal changes in the right insular region without mass effect or contrast enhancement. Electroencephalography revealed intermittent sharp wave activity in this area. Antiepileptic medication was started, and the patient's symptoms resolved. Follow-up MR imaging 6 weeks later revealed enlargement of the lesion and contrast enhancement in the superior temporal gyrus. Consequently, the patient was scheduled for a brain biopsy. However, surgical planning MR images obtained on the day of surgery 4 weeks later showed that the enhancement had spontaneously resolved, and so the operation was cancelled. Repeat MR imaging performed 2 weeks later as a result of increased seizure frequency redemonstrated contrast enhancement. The patient then underwent a craniotomy, and final histopathology was consistent with glioblastoma.


Impact of Optimal Fenestration Size on Outcomes of High-Risk Fontan Patients.
We aimed to analyze mid-term outcomes of the fenestrated Fontan procedure, focusing on the fenestration size. We retrospectively reviewed the outcomes of the fenestrated Fontan procedure. Among 165 patients who underwent the Fontan procedure from 2011 to 2021, fenestration was created in 27 patients with the highest risks, including those with hypoplastic left heart syndrome, hypoplastic pulmonary arteries, heterotaxy syndrome with high pulmonary vascular resistance, and pulmonary arterial pressure >15 mm Hg. The patients underwent the procedure at a median age of three years (body weight, 11.4 kg; body surface area, 0.54 m2). Fenestration sizes were 3.5 to 5 mm. Spontaneous fenestration closure occurred within one year postoperatively in nine patients. Among them, three experienced Fontan failure, necessitating refenestration. Although fenestration size did not differ, the size corrected by body surface area at the time of surgery was smaller in patients with fenestration closure (6.4 mm/m2 vs 8.3 mm/m2, P < .05). Patients with a fenestration <7 mm/m2 were more likely to have fenestration closure within one year postoperatively, and those with a fenestration >9 mm/m2 were more likely to have severe desaturation and require home oxygen therapy following discharge. Spontaneous fenestration closure affected the frequency of Fontan complications. A very small fenestration size corrected by body surface area was a significant risk factor for spontaneous closure. Conversely, a very large fenestration size corrected by body surface area resulted in severe desaturation. The optimal fenestration size to prevent early spontaneous closure and severe desaturation is approximately 8 mm/m2.


Grinding With Diamond Burs and Hydrothermal Aging of a Y-TZP Material: Effect on the Material Surface Characteristics and Bacterial Adhesion.
The aim of this study was to evaluate the effect of grinding with diamond burs and low-temperature aging on the material surface characteristics and bacteria adhesion on a yttrium-stabilized tetragonal zirconia polycrystalline (Y-TZP) surface. Y-TZP specimens were made from presintered blocks, sintered as recommended by the manufacturer, and assigned into six groups according to two factors-grinding (three levels: as sintered, grinding with extra-fine diamond bur [25-μm grit], and grinding with coarse diamond bur [181-μm grit]) and hydrothermal aging-to promote low-temperature degradation (two levels: presence/absence). Phase transformation (X-ray diffractometer), surface roughness, micromorphological patterns (atomic force microscopy), and contact angle (goniometer) were analyzed. Bacterial adhesion (colony-forming units [CFU]/biofilm) was quantified using an in vitro polymicrobial biofilm model. Both the surface treatment and hydrothermal aging promoted an increase in m-phase content. Roughness values increased as a function of increasing bur grit sizes. Grinding with a coarse diamond bur resulted in significantly lower values of contact angle (p<0.05) when compared with the extra-fine and control groups, while there were no differences (p<0.05) after hydrothermal aging simulation. The CFU/biofilm results showed that neither the surface treatment nor hydrothermal aging simulation significantly affected the bacteria adherence (p>0.05). Grinding with diamond burs and hydrothermal aging modified the Y-TZP surface properties; however, these properties had no effect on the amount of bacteria adhesion on the material surface.


Brachyury gene copy number gain and activation of the PI3K/Akt pathway: association with upregulation of oncogenic Brachyury expression in skull base chordoma.
OBJECTIVE Chordoma is a slow-growing but clinically malignant tumor, and the prognosis remains poor in many cases. There is a strong impetus to develop more effective targeted molecular therapies. On this basis, the authors investigated the potential of Brachyury, a transcription factor involved in notochord development, as a candidate molecular target for the treatment of chordoma. METHODS Brachyury gene copy number and expression levels were evaluated by quantitative polymerase chain reaction in 27 chordoma samples, and the transcriptomes of Brachyury high-expression tumors (n = 4) and Brachyury low-expression tumors (n = 4) were analyzed. A chordoma cell line (U-CH2) was used to investigate the signaling pathways that regulate Brachyury expression. RESULTS All chordoma specimens expressed Brachyury, and expression levels varied widely. Patients with higher Brachyury expression had significantly shorter progression-free survival (5 months, n = 11) than those with lower expression (13 months, n = 16) (p = 0.03). Somatic copy number gain was confirmed in 12 of 27 (44%) cases, and copy number was positively correlated with Brachyury expression (R = 0.61, p < 0.001). Expression of PI3K/Akt pathway genes was upregulated in Brachyury high-expression tumors, and suppression of PI3K signaling led to reduced Brachyury expression and inhibition of cell growth in the U-CH2 chordoma cell line. CONCLUSIONS Activation of the PI3K/Akt pathway and Brachyury copy number gain are strongly associated with Brachyury overexpression, which appears to be a key event in chordoma growth regulation. These findings suggest that targeting Brachyury and PI3K/Akt signaling may be an effective new approach for treating chordoma.


Cellular contractility and extracellular matrix stiffness regulate matrix metalloproteinase activity in pancreatic cancer cells.
The pathogenesis of cancer is often driven by local invasion and metastasis. Recently, mechanical properties of the tumor microenvironment have been identified as potent regulators of invasion and metastasis, while matrix metalloproteinases (MMPs) are classically known as significant enhancers of cancer cell migration and invasion. Here we have been able to sensitively measure MMP activity changes in response to specific extracellular matrix (ECM) environments and cell contractility states. Cells of a pancreatic cancer cell line, Panc-1, up-regulate MMP activities between 3- and 10-fold with increased cell contractility. Conversely, they down-regulate MMP activities when contractility is blocked to levels seen with pan-MMP activity inhibitors. Similar, albeit attenuated, responses are seen in other pancreatic cancer cell lines, BxPC-3 and AsPC-1. In addition, MMP activity was modulated by substrate stiffness, collagen gel concentration, and the degree of collagen cross-linking, when cells were plated on collagen gels ranging from 0.5 to 5 mg/ml that span the physiological range of substrate stiffness (50-2000 Pa). Panc-1 cells showed enhanced MMP activity on stiffer substrates, whereas BxPC-3 and AsPC-1 cells showed diminished MMP activity. In addition, eliminating heparan sulfate proteoglycans using heparinase completely abrogated the mechanical induction of MMP activity. These results demonstrate the first functional link between MMP activity, contractility, and ECM stiffness and provide an explanation as to why stiffer environments result in enhanced cell migration and invasion.


Prophylactic co-trimoxazole in biliary surgery.
A double-blind study was conducted to test the prophylactic effect of a single dose of co-trimoxazole on the incidence of septic complications after elective cholecystectomy. Forty-eight patients received co-trimoxazole and 47 placebo. Wound sepsis occurred in 10 (21%) of the controls but in only 2 (4%) of the treated group, and the incidences of pulmonary complications were 49% (23 cases) and 19% (9) respectively. These differences were significnat. Wound sepsis after cholecystectomy occurs mostly in patients with infected bile. Co-trimoxazole given by intravenous infusion rapidly achieves a high concentration in the palsma and is effective against most biliary pathogens.


Writers' award winner experiences of family members acting as primary caregivers for patients with traumatic brain injury.
Research on caregiving experiences of families of patients with traumatic brain injuries has been limited to studies emphasizing negative aspects of care. During this study, families caring for patients with traumatic brain injury (TBI) shared their experiences, allowing us to collect basic data about factors related to their psychosocial adjustment. Fifteen primary caregivers were interviewed using semistructured interviews, and content analysis was performed on obtained data. One thousand eighteen recording units were grouped into seven categories: (1) spending time with the patient with TBI and understanding invisible disabilities, (2) evaluating medical professionals' handling of the case, (3) devoting oneself to caring for the patient with TBI despite feeling psychological distress, (4) accepting the disability and constructing a care system, (5) seeking out and participating in specialized treatment (rehabilitation) for TBIs, (6) hoping for better understanding and creation of an appropriate response system for TBIs, and (7) making efforts to influence people around the caregiver and those in similar circumstances to promote deeper understanding of TBIs.


Sensitivity measures of visuospatial attention.
Measuring visual sensitivity has become popular to determine the spatial deployment of visual attention. Critically, the accuracy of the measurement depends on the quality of the stimulus used. We evaluated the strengths and weaknesses of six commonly used stimuli for assessing visual attention. While preparing an eye movement to a cued item, participants discriminated a stimulus-specific visual feature, either at the cued location or at other equidistant uncued locations. Stimuli differed in their visual features (digital letters, Gabors, crosses, pink noise, random dot kinematograms, and Gabor streams) and their presentation mode (static or dynamic stimuli). We evaluated these stimuli regarding their temporal and spatial specificity and their impact on saccade preparation. We assessed presaccadic visual sensitivity as a correlate of visual spatial attention and discuss the stimulus-specific time course, spatial specificity, and magnitude of the measured attention modulation. Irrespective of the stimulus type, we observed a clear increase of visual sensitivity at the cued location. Time course, spatial specificity, and magnitude of this improvement, however, were specific to each stimulus. Based on our findings, we present guidelines to select the stimulus best suited to measure visuospatial attention depending on the respective research question.


The interaction of fingermark deposits on metal surfaces and potential ways for visualisation.
The interaction of fingermark deposits on metals has been examined by a variety of techniques. Visualisation by film growth has been the main area of investigation through: thermal oxidation, anodising, peroxide solution, and the interaction with vapour of iodine and ammonium sulphide. Corrosion of the underlying metal has also been examined as an alternative means of visualisation. Confocal microscopy was used to look at the film thickness and corrosion products around the prints. Scanning electron microscopy and energy dispersion of X-rays (SEM-EDX) examined a number of metal samples to investigate film growth and the elemental distribution. The observations suggest that differential oxidation was occurring as well as corrosion into the metal. Fingermark deposits on metals can corrode into the metal depending on the reactivity of the metal and leave a recoverable mark. However, fingermark deposits can also alter the rate of chemical reaction of the substrate metal by oxidation. In some cases organic matter can inhibit reaction, both when forming an oxide layer and when corroding the metal. However, signs of third level detail from pore contact may also be visible and the monovalent ions from salts could also influence film growth. Whilst further work would need to be carried out to decide whether any of these techniques may have application in fingermark recovery, this study does suggest that fingermarks on metals may be recoverable after incidents such as fires or immersion in water.


Compact and low loss silicon-on-insulator rib waveguide 90 degrees bend.
A compact and low loss silicon-on-insulator rib waveguide 90 degrees bend is designed and demonstrated. An interface realized by a trench filled with SU8 at the corner of a waveguide bend effectively reflects incoming light through total internal reflection (TIR). In order to accurately position the SU8-filled trench relative to the waveguide and reduce sidewall roughness of the interface, electron beam lithography (EBL) is employed while inductively coupled plasma reactive ion etching (ICP RIE) is used to achieve a vertical sidewall. The measured loss for TE polarization is 0.32 dB +/- 0.02 dB/bend at a wavelength of 1.55 microm.


Glucose positions affect the phloem mobility of glucose-fipronil conjugates.
In our previous work, a glucose-fipronil (GTF) conjugate at the C-1 position was synthesized via click chemistry and a glucose moiety converted a non-phloem-mobile insecticide fipronil into a moderately phloem-mobile insecticide. In the present paper, fipronil was introduced into the C-2, C-3, C-4, and C-6 positions of glucose via click chemistry to obtain four new conjugates and to evaluate the effects of the different glucose isomers on phloem mobility. The phloem mobility of the four new synthetic conjugates and GTF was tested using the Ricinus seedling system. The results confirmed that conjugation of glucose at different positions has a significant influence on the phloem mobility of GTF conjugates.


Serum creatinine is associated with coronary disease risk even in the absence of metabolic disorders.
In view of recent evidence that serum creatinine and dysfunctional apolipoprotein (apo)A-I may serve as inflammation mediators in people with enhanced inflammation, we studied whether or not these molecules were interrelated and associated with coronary heart disease (CHD) likelihood even in subjects without metabolic syndrome (MetS) or type-2 diabetes. Among unselected middle-aged Turkish adults with available serum apo A-I, lipoprotein(a) and creatinine measurements, 697 participants (designated as 'healthy') were enrolled, after exclusion of the stated metabolic disorders. CHD was identified in 87 subjects, roughly half during 3.1 years' follow-up. 'Healthy' individuals were overweight and had partly impaired fasting glucose but otherwise normal serum creatinine and other biochemical measurements. Being consistent with lacking anti-inflammatory activity, apoA-I was linearly and positively associated with apoB, in women further with creatinine. Logistic regression analyses showed that, beyond age, not non-HDL-cholesterol, systolic blood pressure and smoking status, but serum creatinine in each sex (OR in men 1.63 [95% CI 1.14; 2.31]) and CRP in women were significantly associated with CHD likelihood. The combined highest and lowest creatinine quartiles in women displayed an OR 2.14 (1.02; 4.51) compared with the intermediate quartiles, after similar adjustments. Elevated creatinine levels within normal range, linked to apoA-I dysfunctionality, are independently associated with CHD likelihood even in non-diabetic subjects without MetS. In such women the lowest creatinine quartile is also linked to CHD risk.


Elevation of cAMP in mesenchymal stem cells transiently upregulates neural markers rather than inducing neural differentiation.
The aims of this research were to examine the neural expression profile of undifferentiated mesenchymal stem cells (MSCs), to define a serum-free environment that would support the survival of MSCs, and to assess the effects of elevated cyclic adenosine monophosphate (cAMP) levels on MSC morphology and expression of neural markers. The neural profile of MSCs was assessed using immunocytochemistry and real-time polymerase chain reaction (PCR) techniques. These MSCs were then cultured in varying serum-free environments, with flow cytometry analysis of cell viability and apoptosis. The effects of forskolin and 8 bromo-cAMP treatment on MSC morphology and expression of neural markers were assessed using light microscopy, immunocytochemistry and real-time PCR analysis. Expression of the neural markers nestin, vimentin, beta III tubulin, glial fibrillary acidic protein, and the tropomyosin-related kinases was demonstrated in normal undifferentiated MSCs. Furthermore, MSCs cultured in serum-free conditions containing ascorbic acid 2-phosphate demonstrated greater long-term survival and reduced apoptosis. In contrast, forskolin increased the percentage of cells undergoing apoptosis. Culture in the presence of forskolin induced a 6-fold increase in beta III tubulin expression after a 6-h exposure time, which was accompanied by dramatic changes in morphology. However, this effect was transient, with the cells reverting to their normal morphology by 24 h. Treatment with 8 bromo-cAMP induced similar increases in beta III tubulin expression without such dramatic morphological changes. Factors which increase the concentration of cAMP induce transient changes in expression of neural markers that appear to be the result of cellular adaptation to changes in culture conditions rather than a real process of neural differentiation.


[Bacterial keratitis. Microbiological analysis as a principle for therapeutic recommendations].
The study presented differentiates between the aetiological agents of bacterial keratitis in patients with and without a history of contact lens wear. Based on these results, recommendations are given for optimal antibiotic primary therapy. Swabs and corneal scrapings were taken from 218 patients referred to the University Eye Hospital in Munich with a diagnosis of bacterial keratitis from 1989 to 1997. Ninety-two of these patients had a history of contact lens wear; 126 had none. The germs were isolated and identified by staining and microscopy. Observing polymicrobial growth in 51 patients, a total of 275 germs was isolated. The most frequent pathogens were Staphylococcus epidermidis (44%), S. aureus (18%), Streptococcus spp. (10%), Propionibacterium acnes (7%) and Pseudomonas aeruginosa (6%). Gram-negative germs were nearly exclusively isolated from contact lens wearers, gram-positive germs were predominant in non-contact lens wearers. Keratitis in patients with a history of contact lens wear is often caused by aggressive gram-negative germs. Those cases should immediately be treated with quinolones and erythromycin. In keratitis caused by gram-positive pathogens, a combination with aminoglycosides and erythromycin is sufficient.


A very fast algorithm for simultaneously performing connected-component labeling and euler number computing.
Labeling connected components and calculating the Euler number in a binary image are two fundamental processes for computer vision and pattern recognition. This paper presents an ingenious method for identifying a hole in a binary image in the first scan of connected-component labeling. Our algorithm can perform connected component labeling and Euler number computing simultaneously, and it can also calculate the connected component (object) number and the hole number efficiently. The additional cost for calculating the hole number is only O(H) , where H is the hole number in the image. Our algorithm can be implemented almost in the same way as a conventional equivalent-label-set-based connected-component labeling algorithm. We prove the correctness of our algorithm and use experimental results for various kinds of images to demonstrate the power of our algorithm.


Kayexalate Intake (in Sorbitol) and Jejunal Diverticulitis, a Causative Role or an Innocent Bystander?
Small intestine diverticulosis is a rare entity that is asymptomatic in the majority of cases. However, it may cause serious complications, such as infection, hemorrhage, intestinal obstruction and diverticulitis. Kayexalate (sodium polystyrene sulfonate) in sorbitol has been associated with colonic necrosis and less frequently with upper gastrointestinal injuries in a subset of uremic patients treated for hyperkalemia. We report a case of jejunal diverticulosis with mucosal injury and diverticulitis in a uremic patient treated with Kayexalate and discuss the potential role of Kayexalate in the pathogenesis of diverticulitis.


Preload Responsiveness in Patients With Acute Respiratory Distress Syndrome Managed With Extracorporeal Membrane Oxygenation.
A restrictive fluid strategy is recommended in patients with acute respiratory distress syndrome (ARDS) managed with venovenous extracorporeal membrane oxygenation (VV ECMO). However, there are no established predictors for preload responsiveness in these patients. In 20 ARDS patients managed with VV ECMO, transesophageal echocardiography was used to repeatedly evaluate dynamic parameters of the left (velocity and stroke volume variation) and right ventricular outflow tract (velocity [respiratory variations of the maximal Doppler velocity in the truncus pulmonalis {ΔVmaxTP}] and velocity time integral [respiratory variation of the velocity time integral measured in the truncus pulmonalis {ΔVTI_TP}] variation in the truncus pulmonalis), the diameter variation in the superior and inferior vena cava and stroke volume variation measured by pulse contour analysis (SVV_PCA). Patients were categorized as responders and nonresponders according to an increase in stroke volume measured by echocardiography during a Passive Leg Raise Test with a cutoff value ≥10%. The final analysis includes 86 measurements. Predictive values for preload responsiveness were found for ΔVmaxTP (area under the curve [AUC] of 0.64), ΔVTI_TP (AUC 0.67), and SVV_PCA (AUC 0.74). In conclusion, SVV_PCA and, to a lesser extent, ΔVmaxTP and ΔVTI_TP are the most accurate parameters to predict preload responsiveness in ARDS patients managed with VV ECMO. Transesophageal echocardiography offers no advantages over pulse contour analysis for predicting preload responsiveness and provides only intermittent monitoring and assessment.


3-Monoiodothyronamine: the rationale for its action as an endogenous adrenergic-blocking neuromodulator.
The investigations reported here were designed to gain insights into the role of 3-monoiodothyronamine (T1AM) in the brain, where the amine was originally identified and characterized. Extensive deiodinase studies indicated that T1AM was derived from the T4 metabolite, reverse triiodothyronine (revT3), while functional studies provided well-confirmed evidence that T1AM has strong adrenergic-blocking effects. Because a state of adrenergic overactivity prevails when triiodothyronine (T3) concentrations become excessive, the possibility that T3's metabolic partner, revT3, might give rise to an antagonist of those T3 actions was thought to be reasonable. All T1AM studies thus far have required use of pharmacological doses. Therefore we considered that choosing a physiological site of action was a priority and focused on the locus coeruleus (LC), the major noradrenergic control center in the brain. Site-directed injections of T1AM into the LC elicited a significant, dose-dependent neuronal firing rate change in a subset of adrenergic neurons with an EC(50)=2.7 microM, a dose well within the physiological range. Further evidence for its physiological actions came from autoradiographic images obtained following intravenous carrier-free (125)I-labeled T1AM injection. These showed that the amine bound with high affinity to the LC and to other selected brain nuclei, each of which is both an LC target and a known T3 binding site. This new evidence points to a physiological role for T1AM as an endogenous adrenergic-blocking neuromodulator in the central noradrenergic system.


Insoluble/soluble fraction ratio determines effects of dietary fiber on gut microbiota and serum metabolites in healthy mice.
Both soluble dietary fiber (SDF) and insoluble dietary fiber (IDF) play pivotal roles in maintaining gut microbiota homeostasis; whether the effects of the different ratios of IDF and SDF are consistent remains unclear. Consequently, we selected SDFs and IDFs from six representative foods (apple, celery, kale, black fungus, oats, and soybeans) and formulated nine dietary fiber recipes composed of IDF and SDF with a ratio from 1 : 9 to 9 : 1 (NDFR) to compare their impact on microbial effects with healthy mice. We discovered that NDFR treatment decreased the abundance of Proteobacteria and the ratio of Firmicutes/Bacteroidetes at the phylum level. The α diversity and relative richness of Parabacteroides and Prevotella at the genus level showed an upward trend along with the ratio of IDF increasing, while the relative abundance of Akkermansia at the genus level and the production of acetic acid and propionic acid exhibited an increased trend along with the ratio of SDF increasing. The relative abundance of Parabacteroides and Prevotella in the I9S1DF group (the ratio of IDF and SDF was 9 : 1) was 1.72 times and 5.92 times higher than that in the I1S9DF group (the ratio of IDF and SDF was 1 : 9), respectively. The relative abundance of Akkermansia in the I1S9DF group was 17.18 times higher than that in the I9S1DF group. Moreover, a high ratio of SDF (SDF reaches 60% or more) enriched the glycerophospholipid metabolism pathway; however, a high ratio of IDF (IDF reaches 80% or more) regulated the tricarboxylic acid cycle. These findings are helpful in the development of dietary fiber supplements based on gut microbiota and metabolites.


Broad and potently neutralizing monoclonal antibodies isolated from human survivors of New World hantavirus infection.
New World hantaviruses (NWHs) are endemic in North and South America and cause hantavirus cardiopulmonary syndrome (HCPS), with a case fatality rate of up to 40%. Knowledge of the natural humoral immune response to NWH infection is limited. Here, we describe human monoclonal antibodies (mAbs) isolated from individuals previously infected with Sin Nombre virus (SNV) or Andes virus (ANDV). Most SNV-reactive antibodies show broad recognition and cross-neutralization of both New and Old World hantaviruses, while many ANDV-reactive antibodies show activity for ANDV only. mAbs ANDV-44 and SNV-53 compete for binding to a distinct site on the ANDV surface glycoprotein and show potently neutralizing activity to New and Old World hantaviruses. Four mAbs show therapeutic efficacy at clinically relevant doses in hamsters. These studies reveal a convergent and potently neutralizing human antibody response to NWHs and suggest therapeutic potential for human mAbs against HCPS.


Long-term antitrypanosomal effect of quinapyramine sulphate-loaded oil-based nanosuspension in T. evansi-infected mouse model.
The goal of the present work consisted of the formulation development and evaluation of quinapyramine sulphate (QS)-loaded long-acting oil-based nanosuspension for improved antitrypanosomal effect. QS was transformed into a hydrophobic ionic complex using anionic sodium cholate (Na.C). The complex was characterized by FTIR, DSC, and XRD. Oil-based nanosuspension was prepared by dispersing the QS-Na.C complex in thixotropically thickened olive oil. The nanoformulation was found to be cytocompatible (82.5 ± 5.87% cell viability at the minimum effective concentration [MEC]) in THP-1 cell lines and selectively trypanotoxic (p < 0.0001). The pharmacokinetic studies of QS-Na.C complex-loaded oily nanosuspension showed 13.54-fold, 7.09-fold, 1.78-fold, and 17.35-fold increases in t1/2, AUC0-∞, Vz/F, and MRT0-ꝏ, respectively, as compared to free QS. Moreover, a 7.08-fold reduction in plasma clearance was observed after the treatment with the optimized formulation in Wistar rats. Furthermore, treatment with QS-Na.C complex-loaded oily nanosuspension (7.5 mg/kg) in T. evansi-infected mice model showed the absence of parasitaemia for more than 75 days after the treatment during in vivo efficacy studies. The efficacy of the treatment was assessed by observation of blood smear and PCR assay for DNA amplification. To conclude, our findings suggest that the efficient delivery of QS from the developed QS-Na.C complex-loaded oily nanosuspension could be a promising treatment option for veterinary infections against trypanosomiasis.


Carers' Experiences, Needs, and Preferences During Inpatient Stroke Rehabilitation: A Systematic Review of Qualitative Studies.
To report and synthesize the experiences, needs, and preferences of carers of stroke survivors undergoing inpatient rehabilitation. MEDLINE, CINAHL, Embase, PsycINFO, and Web of Science were searched to March 2016. Reference lists of relevant publications were searched. No language restrictions were applied. Eligible qualitative studies reported the experiences of carers of stroke survivors who underwent inpatient rehabilitation. The search yielded 3532 records; 93 full-text publications were assessed for eligibility, and 34 documents (33 studies) were included. Comprehensiveness of reporting was assessed using the Consolidated Criteria for Reporting Qualitative Health Research framework. Data on the characteristics of included studies were independently extracted by 2 authors. Differences in data extraction between authors were resolved through discussion or by a third author. All text in studies' results and discussion sections were extracted for analysis. Extracted texts were analyzed inductively using thematic synthesis. Seven analytical themes were developed that related to the carers' experiences, needs, and preferences: (1) overwhelmed with emotions; (2) recognition as a stakeholder in recovery; (3) desire to be heard and informed; (4) persisting for action and outcomes; (5) being legitimate clients; (6) navigating an alien culture and environment; and (7) managing the transition home. This systematic review provides new insights into the experiences, needs, and preferences of carers of stroke survivors undergoing inpatient rehabilitation. Carers experienced distress as they navigated a foreign culture and environment without adequate communication and processes in place for their inclusion. We recommend deliberate efforts to provide a more inclusive environment that better supports and prepares carers for their new role.


[Epilepsy and sleep disorders].
Sleep disorders often occur in patients with epilepsy. Every neurologist is familiar with the postictal drowsiness after tonicoclonic convulsions and likewise with the provocation of an attack after sleep deprivation that is often combined with alcohol consumption. It is more difficult to differentiate between motor disturbances and epileptic episodes during sleep. For example, nocturnal paroxysmal dystonia, which are frequently an expression of frontal lobe epilepsy, are long not recognized as non-epileptic, sleep-associated attacks. In addition, nocturnal episodes in context of a REM sleep behavioral disturbance can occasionally be differentiated from frontal lobe epileptic seizures only with great difficulty. A clear differential diagnostics between epileptic and non-epileptic attacks during sleep is, however, a requirement for a selective treatment.


Identifying almost invariant sets in stochastic dynamical systems.
We consider the approximation of fluctuation induced almost invariant sets arising from stochastic dynamical systems. The dynamical evolution of densities is derived from the stochastic Frobenius-Perron operator. Given a stochastic kernel with a known distribution, approximate almost invariant sets are found by translating the problem into an eigenvalue problem derived from reversible Markov processes. Analytic and computational examples of the methods are used to illustrate the technique, and are shown to reveal the probability transport between almost invariant sets in nonlinear stochastic systems. Both small and large noise cases are considered.


Iron-sulfur stoichiometry and structure of iron-sulfur clusters in three-iron proteins: evidence for [3Fe-4S] clusters.
Beef heart aconitase contains 3Fe clusters in its inactive and 4Fe clusters in its active form. The fully active form can be restored from the inactive one by insertion of Fe(2+), whereas S(2-) is not required. Chemical analyses for iron and labile sulfide yield Fe/S(2-) ratios of 0.66-0.74 for the inactive and 0.90-1.03 for the active form. Sulfane sulfur (S(0)) was not detected. We propose on the basis of these data that the inactive form may arise from the active one by loss of one iron only per cluster with the sulfur remaining as S(2-) in a [3Fe-4S] structure. Measurements by extended x-ray absorption fine structure (EXAFS) spectroscopy on the 3Fe form of aconitase yield a Fe..S distance of 2.24 A and a Fe..Fe distance of 2.71 A. This Fe..Fe distance is in agreement with that obtained by EXAFS on ferredoxin II of Desulfovibrio gigas, another 3Fe protein, but disagrees with Fe..Fe distances observed for the 3Fe cluster of Azotobacter vinelandii ferredoxin I by x-ray diffraction-namely, 4.1 A. We suggest that this difference may be due to the presence of a [3Fe-3S] structure in the Azotobacter ferredoxin I crystals vs. a [3Fe-4S] structure in liquid or frozen solutions of aconitase. The [3Fe-3S] cluster has been shown to have a relatively flat twist-boat structure, whereas a [3Fe-4S] cluster could be expected to essentially maintain the compact structure of the [4Fe-4S] cluster. This would explain the differences in Fe..Fe distances. Two possible structural models for a [3Fe-4S] cluster are discussed.


Analysis of treatment adherence in patients undergoing phototherapy.
Phototherapy is a convenient and effective treatment alternative for a range of skin diseases. However, a major challenge in patient adherence to phototherapy may be the necessity of visiting a phototherapy center regularly over an extended period of time. The aims of this study were (i) to investigate the adherence rate to phototherapy and (ii) to determine factors associated with adherence to narrow-band ultraviolet B phototherapy (nbUVB) treatment. A retrospective review of patient records who underwent nbUVB phototherapy between January 1, 2018, and March 31, 2023, was performed. Patient records were reviewed for age, gender, skin type, diagnosis, type of phototherapy applied, duration of treatment, total number of sessions, presence of side effects, reasons for discontinuation of treatment, and perceived benefits of treatment. Of a total of 729 patients undergoing phototherapy, 281 (38.5%) discontinued treatment before completing 20 sessions. In particular, younger patients and those who experienced fewer side effects tended to discontinue treatment prematurely. The most common reason for discontinuing treatment was difficulty in visiting the hospital regularly. The patient's compliance with phototherapy was 61.5%. These results indicate that phototherapy is still one of the preferred treatment methods, although many new treatment agents have been developed in dermatology in recent years. Identifying and addressing factors that affect patient adherence will certainly help increase the effectiveness of treatment.


NRG1/ERBB3 Pathway Activation Induces Acquired Resistance to XPO1 Inhibitors.
XPO1 inhibitors have shown promise in cancer treatment, but mechanisms of resistance to these drugs are not well understood. In this study, we established selective inhibitors of nuclear export (SINE)-resistant ovarian cancer cell lines from in vivo mouse tumors and determined the mechanisms of adaptive XPO1 inhibitor resistance using protein and genomic arrays. Pathway analyses revealed upregulation of the NRG1/ERBB3 pathway in SINE-resistant cells. Depletion of ERBB3 using siRNAs restored the antitumor effect of SINE in vitro and in vivo Furthermore, exogenous NRG1 decreased the antitumor effect of SINE in ovarian cancer cell lines with high ERBB3 expression, but not in those with low expression. These results suggest that NRG1 and ERBB3 expression is a potential biomarker of response to SINE treatment. The antitumor effect of SINE was reduced by exogenous NRG1 in an ERBB3-dependent manner. These findings suggest that NRG1 and ERBB3 are effective biomarkers that should be evaluated in future clinical trials and are relevant therapeutic targets for the treatment of SINE-resistant cancers.


Electrical synapses in the mammalian brain.
Many neurons in the mammalian central nervous system communicate through electrical synapses, defined here as gap junction-mediated connections. Electrical synapses are reciprocal pathways for ionic current and small organic molecules. They are often strong enough to mediate close synchronization of subthreshold and spiking activity among clusters of neurons. The most thoroughly studied electrical synapses occur between excitatory projection neurons of the inferior olivary nucleus and between inhibitory interneurons of the neocortex, hippocampus, and thalamus. All these synapses require the gap junction protein connexin36 (Cx36) for robust electrical coupling. Cx36 appears to interconnect neurons exclusively, and it is expressed widely along the mammalian neuraxis, implying that there are undiscovered electrical synapses throughout the central nervous system. Some central neurons may be electrically coupled by other connexin types or by pannexins, a newly described family of gap junction proteins. Electrical synapses are a ubiquitous yet underappreciated feature of neural circuits in the mammalian brain.


Characterization of MPF activation by okadaic acid in Xenopus oocyte.
Okadaic acid (OA), a specific inhibitor of protein phosphatases, induces a rapid activation (30 min) of MPF when microinjected into the Xenopus oocyte. Neither protein synthesis inhibitors nor cAMP counteract the action of OA. These results indicate that the inhibition of protein phosphatase(s) is sufficient for the in vivo activation of MPF even after the full activation of cAMP-dependent protein kinase. In all experimental conditions (plus or minus inhibitors of protein synthesis; normal or elevated cAMP levels) OA induces a burst of protein phosphorylation together with the activation of MPF. Cytological analysis shows that OA provokes the breakdown of the nuclear envelope, the depolymerization of lamin and the condensation of the chromosomes. However, no metaphase spindles are organized, indicating that inhibition of protein phosphatases strongly affects the function of the microtubule organizing center.


Ammonium transport in the kidney: new physiological concepts and their clinical implications.
This article is based on a Basic Science Symposium presented at the 23rd Annual Meeting of the American Society of Nephrology. New information on the segmental transport of ammonium by the proximal tubule, the thick ascending limb of Henle's loop, and the collecting duct is integrated into a thesis that NH4+ excretion is controlled by the rate of production, by diffusion of NH3 along gradients established by proton secretion, and by active transport of NH4+. These new concepts are applied to a novel explanation of the pathogenesis of distal renal tubular acidosis.


Alternative TrkAIII splicing: a potential regulated tumor-promoting switch and therapeutic target in neuroblastoma.
An association between elevated tyrosine kinase receptor (Trk)-A expression and better prognosis; the absence of mutation-activated TrkA oncogenes; the induction of apoptosis, growth arrest, morphological differentiation and inhibition of xenograft growth; and angiogenesis by TrkA gene transduction, provide the basis for the current concept of an exclusively tumor-suppressor role for TrkA in the aggressive pediatric tumor, neuroblastoma. This concept, however, has recently been challenged by the discovery of a novel hypoxia-regulated alternative TrkAIII splice variant, initial data for which suggest predominant expression in advanced-stage neuroblastoma. TrkAIII exhibits neuroblastoma xenograft tumor-promoting activity associated with the induction of a more angiogenic and stress-resistant neuroblastoma phenotype and antagonises nerve growth factor/TrkAI antioncogenic signaling. In this short review, the authors integrate this novel information into a modified concept that places alternative TrkA splicing as a potential pivotal regulator of neuroblastoma behavior and identifies the TrkAIII alternative splice variant as a potential biomarker of patient prognosis and novel therapeutic target.


A membrane bioreactor for the removal of dimethyl sulphide and toluene from waste air.
In biotrickling filters, mass transfer of hydrophobic compounds is the limiting factor. Biofilters are static systems, and so control and regulation of operational parameters such as pH and nutrient supply can be a problem. In membrane bioreactors, these drawbacks can be avoided. The hydrophobic membrane separates the waste air from the aqueous phase, thus avoiding mass-transfer limitation, while pH and nutrient supply can be directly controlled. In this contribution, an overview will be given of results obtained during a four-year project. First, the physical chemical characteristics (solubility, permeability, diffusivity) and microbial adhesion of different membranes were tested. This led to the selection of a composite membrane consisting of a porous polyvinylidenefluoride (PVDF) support layer coated with a thin (1 or 2.5 microm) dense polydimethylsiloxane (PDMS) top layer. This membrane was mounted into a module provided with four parallel rectangular channels for gas flow (in contact with the porous layer) and nutrient solution (in contact with the dense layer) respectively. After inoculation, a biofilm developed on the dense layer. Experiments were performed with dimethyl sulphide and toluene as target VOCs. Operational characteristics such as elimination capacity as a function of the volumetric load and residence time, effect of nutrient supply, long-term performance) were determined. Mass transfer was studied by measuring concentration profiles along the channels of the module in different conditions.


A role for MCH neuron firing in modulating hippocampal plasticity threshold.
It has been revealed that hypothalamic neurons containing the peptide, melanin-concentrating hormone (MCH) can influence learning [1] and memory formation [2], but the cellular mechanisms by which they perform this function are not well understood. Here, we examine the role of MCH neural input to the hippocampus, and show in vitro that optogenetically increasing MCH axon activity facilitates hippocampal plasticity by lowering the threshold for synaptic potentiation. These results align with increasing evidence that MCH neurons play a regulatory role in learning, and reveal that this could be achieved by modulating plasticity thresholds in the hippocampus.


Women's help-seeking patterns for depression.
Depression tends not to be accurately identified and treated in primary care settings. Women of color are especially likely to use these settings for mental health issues but are less likely to be diagnosed appropriately. A study was conducted within four Florida primary care clinics serving primarily low-income families. Participants included 321 women (Black, 22%, Hispanic, 23.5%, White, 38.6%) who completed a confidential questionnaire while waiting to be seen by clinic staff. Ten percent reported recent major depression, with 26.7% indicating depressive symptoms during the past two years. Depressed women were significantly more likely to report physical violence during the past year. Respondents turned primarily to family, friends, and medical clinics for their depression. They found turning to clinic staff to be helpful and described psychosocial interventions as useful. Members of all ethnic/racial groups reported barriers to seeking help, including perceived separation between mental health and general health and stigma. Implications are discussed in terms of appropriate community education and screening procedures.


At risk of population decline? An ecological and genetic approach to the threatened palm species Butia eriospatha (Arecaceae) of Southern Brazil.
To estimate the risk of population decline for the threatened palm species Butia eriospatha, we investigated the patterns of demography, natural regeneration, herbivory, and the levels of genetic diversity using 9 microsatellite loci from both adults and seedlings sampled from 4 populations in Southern Brazil (n = 330). Our results indicate that cattle grazing in B. eriospatha population areas severely affect their demographic structure. Three B. eriospatha populations showed a bimodal age structure made up of adult plants and seedlings and high rates (>77%) of livestock herbivory. For 1 population, we describe and quantify for the first time the occurrence of 6 ontogenetic stages for this threatened palm species. Populations of B. eriospatha showed high levels of genetic differentiation (F ST adult plants = 0.287, F ST seedlings = 0.175). The amount of observed heterozygosity differed significantly between small (H O = 0.329) and large populations (H O = 0.461), indicating that small populations can be more susceptible to genetic drift. With no recruitment and a mortality rate of 2.0%, we show that the populations investigated in this study would be at an extremely high risk of local extinction, with a greater than 50% reduction in the effective population size, in the next 40 years. Although this study highlights the importance of analyzing both population ecology parameters and genetic data to better understand the level of risk facing threatened species, we emphasize that policy actions are urgently needed for effective conservation of this vulnerable biological resource.


A MIF haplotype is associated with the outcome of patients with severe sepsis: a case control study.
Macrophage migration inhibitory factor (MIF) plays an important regulatory role in sepsis. In the promoter region a C/G single nucleotide polymorphism (SNP) at position -173 (rs755622) and a CATT5-8 microsatellite at position -794 are related to modified promoter activity. The purpose of the study was to analyze their association with the incidence and outcome of severe sepsis. Genotype distributions and allele frequencies in 169 patients with severe sepsis, 94 healthy blood donors and 183 postoperative patients without signs of infection or inflammation were analyzed by real time PCR and Sequence analysis. All included individuals were Caucasians. Genotype distribution and allele frequencies of severe sepsis patients were comparable to both control groups. However, the genotype and allele frequencies of both polymorphisms were associated significantly with the outcome of severe sepsis. The highest risk of dying from severe sepsis was detectable in patients carrying a haplotype with the alleles -173 C and CATT7 (p = 0.0005, fisher exact test, RR = 1,806, CI: 1.337 to 2.439). The haplotype with the combination of the -173 C allele and the -794 CATT7 allele may not serve as a marker for susceptibility to sepsis, but may help identify septic patients at risk of dying.


Rhamnose synthase activity is required for pathogenicity of the vascular wilt fungus Verticillium dahliae.
The initial interaction of a pathogenic fungus with its host is complex and involves numerous metabolic pathways and regulatory proteins. Considerable attention has been devoted to proteins that play a crucial role in these interactions, with an emphasis on so-called effector molecules that are secreted by the invading microbe to establish the symbiosis. However, the contribution of other types of molecules, such as glycans, is less well appreciated. Here, we present a random genetic screen that enabled us to identify 58 novel candidate genes that are involved in the pathogenic potential of the fungal pathogen Verticillium dahliae, which causes vascular wilt diseases in over 200 dicotyledonous plant species, including economically important crops. One of the candidate genes that was identified concerns a putative biosynthetic gene involved in nucleotide sugar precursor formation, as it encodes a putative nucleotide-rhamnose synthase/epimerase-reductase (NRS/ER). This enzyme has homology to bacterial enzymes involved in the biosynthesis of the nucleotide sugar deoxy-thymidine diphosphate (dTDP)-rhamnose, a precursor of L-rhamnose, which has been shown to be required for virulence in several human pathogenic bacteria. Rhamnose is known to be a minor cell wall glycan in fungi and has therefore not been suspected as a crucial molecule in fungal-host interactions. Nevertheless, our study shows that deletion of the VdNRS/ER gene from the V. dahliae genome results in complete loss of pathogenicity on tomato and Nicotiana benthamiana plants, whereas vegetative growth and sporulation are not affected. We demonstrate that VdNRS/ER is a functional enzyme in the biosynthesis of uridine diphosphate (UDP)-rhamnose, and further analysis has revealed that VdNRS/ER deletion strains are impaired in the colonization of tomato roots. Collectively, our results demonstrate that rhamnose, although only a minor cell wall component, is essential for the pathogenicity of V. dahliae.


Action of acid on oligoribonucleotide phosphotriester intermediates. Effect of released vicinal hydroxy functions.
When 2'-O-methoxytetrahydropyranyl-5'-O-(9-phenylxanthen-9-yl) uridylyl-(3'----5')-(2',3'-di-O-acetyluridine) 2-chlorophenyl ester (9) is treated with zinc bromide in dichloromethane-propan-2-ol (85:15 v/v) at room temperature, under stringently anhydrous conditions, the corresponding 5'-unblocked dinucleoside phosphate (10) is obtained in 86% isolated yield; however, when no special precautions are taken to exclude moisture, (10) is obtained in only 72% yield. The removal of the 5'-O-(9-phenylxanthen-9-yl) protecting group from (10) with a protic acid (phenyl dihydrogen phosphate) appears to be much less selective and efficient. 80% Acetic acid promoted removal of the methoxytetrahydropyranyl protecting group from the isomeric fully-protected uridylyl-(3'----5')- and uridylyl-(2'----5')-uridine derivatives [(11) and (21c), respectively] leads to virtually identical mixtures [Figures 1a and 1b, respectively] of the partially-protected dinucleoside phosphates [(14) and (15)], 2',3'-di-O-acetyluridine (8), 5'-O-acetyluridine 2',3'-cyclic phosphate (16), and 5'-O-acetyluridine 2'(3')-phosphates [(18) and (17)].


Roles for the Synechococcus elongatus RNA-Binding Protein Rbp2 in Regulating the Circadian Clock.
The cyanobacterial circadian oscillator, consisting of KaiA, KaiB, and KaiC proteins, drives global rhythms of gene expression and compaction of the chromosome and regulates the timing of cell division and natural transformation. While the KaiABC posttranslational oscillator can be reconstituted in vitro, the Kai-based oscillator is subject to several layers of regulation in vivo. Specifically, the oscillator proteins undergo changes in their subcellular localization patterns, where KaiA and KaiC are diffuse throughout the cell during the day and localized as a focus at or near the pole of the cell at night. Here, we report that the CI domain of KaiC, when in a hexameric state, is sufficient to target KaiC to the pole. Moreover, increased ATPase activity of KaiC correlates with enhanced polar localization. We identified proteins associated with KaiC in either a localized or diffuse state. We found that loss of Rbp2, found to be associated with localized KaiC, results in decreased incidence of KaiC localization and long-period circadian phenotypes. Rbp2 is an RNA-binding protein, and it appears that RNA-binding activity of Rbp2 is required to execute clock functions. These findings uncover previously unrecognized roles for Rbp2 in regulating the circadian clock and suggest that the proper localization of KaiC is required for a fully functional clock in vivo.


Comparison of torsional properties between a Fixateur Externe du Service de Santé des Armées and an acrylic tie-in external skeletal fixator in a red-tailed hawk (Buteo jamaicensis) synthetic tibiotarsal bone model.
To compare the torsional mechanical properties of 2 external skeletal fixators (ESFs) placed with 2 intramedullary pin (IP) and transfixation pin (TP) size combinations in a model of raptor tibiotarsal bone fracture. 24 ESF-synthetic tibiotarsal bone model (polyoxymethylene) constructs. Synthetic bone models were fabricated with an 8-mm (simulated fracture) gap. Four types of ESF-synthetic bone model constructs (6/group) were tested: a FESSA with a 1.6-mm IP and 1.6-mm TPs, a FESSA with a 2.0-mm IP and 1.1-mm TPs, an acrylic connecting bar with a 1.6-mm IP and 1.6-mm TPs, and an acrylic connecting bar with a 2.0-mm IP and 1.1-mm TPs. Models were rotated in torsion (5°/s) to failure or the machine angle limit (80°). Mechanical variables at yield and at failure were determined from load deformation curves. Effects of overall construct type, connecting bar type, and IP and TP size combination on mechanical properties were assessed with mixed-model ANOVAs. Both FESSA constructs had significantly greater median stiffness and median torque at yield than both acrylic bar constructs; FESSA constructs with a 1.6-mm IP and 1.6-mm TPs had greatest stiffness of all tested constructs and lowest gap strain at yield. No FESSA constructs failed during testing; 7 of 12 acrylic bar constructs failed by fracture of the connecting bar at the interface with a TP. Although acrylic bar ESFs have been successfully used in avian patients, the FESSA constructs in this study were mechanically superior to acrylic bar constructs, with greatest benefit resulting from use with the larger TP configuration.


Essential genes from Arctic bacteria used to construct stable, temperature-sensitive bacterial vaccines.
All bacteria share a set of evolutionarily conserved essential genes that encode products that are required for viability. The great diversity of environments that bacteria inhabit, including environments at extreme temperatures, place adaptive pressure on essential genes. We sought to use this evolutionary diversity of essential genes to engineer bacterial pathogens to be stably temperature-sensitive, and thus useful as live vaccines. We isolated essential genes from bacteria found in the Arctic and substituted them for their counterparts into pathogens of mammals. We found that substitution of nine different essential genes from psychrophilic (cold-loving) bacteria into mammalian pathogenic bacteria resulted in strains that died below their normal-temperature growth limits. Substitution of three different psychrophilic gene orthologs of ligA, which encode NAD-dependent DNA ligase, resulted in bacterial strains that died at 33, 35, and 37 degrees C. One ligA gene was shown to render Francisella tularensis, Salmonella enterica, and Mycobacterium smegmatis temperature-sensitive, demonstrating that this gene functions in both Gram-negative and Gram-positive lineage bacteria. Three temperature-sensitive F. tularensis strains were shown to induce protective immunity after vaccination at a cool body site. About half of the genes that could be tested were unable to mutate to temperature-resistant forms at detectable levels. These results show that psychrophilic essential genes can be used to create a unique class of bacterial temperature-sensitive vaccines for important human pathogens, such as S. enterica and Mycobacterium tuberculosis.


Preadoptive factors predicting lesbian, gay, and heterosexual couples' relationship quality across the transition to adoptive parenthood.
The authors examined preadoptive factors as predictors of relationship quality (love, ambivalence, and conflict) among 125 couples (44 lesbian couples, 30 gay male couples, and 51 heterosexual couples) across the 1st year of adoptive parenthood. On average, all new parents experienced declines in their relationship quality across the 1st year of parenthood regardless of sexual orientation, with women experiencing steeper declines in love. Parents who, preadoption, reported higher levels of depression, greater use of avoidant coping, lower levels of relationship maintenance behaviors, and less satisfaction with their adoption agencies reported lower relationship quality at the time of the adoption. The effect of avoidant coping on relationship quality varied by gender. Parents who, preadoption, reported higher levels of depression, greater use of confrontative coping, and higher levels of relationship maintenance behaviors reported greater declines in relationship quality. These findings have implications for professionals who work with adoptive parents both pre- and postadoption.


Effects of induced clinical mastitis during preovulation on endocrine and follicular function.
The objective of the study was to determine if experimentally induced clinical mastitis before ovulation resulted in alterations of endocrine function, follicular growth, or ovulation. On d 8 (estrus = d 0), cows were challenged (TRT; n = 19) with Streptococcus uberis or were not challenged (control; n = 14). Forty-eight hours after induction of luteal regression on d 12, blood samples were collected to determine estradiol-17beta, LH pulse frequency, and occurrence of the LH surge. Ovaries were scanned to monitor follicular growth and ovulation. Cows with clinical mastitis (n = 12) had elevated rectal temperatures, somatic cell counts, and mammary scores. Estrus and ovulation occurred in 4 of 12 clinically infected cows and in all control cows. Cows that were challenged but did not develop clinical mastitis (n = 5) displayed estrus and ovulated. Due to differences in expression of estrus, cows were further subdivided for analyses into 4 groups: control, TRT-EST (infected cows that displayed estrus; n = 4), TRT-NOEST (infected cows that did not display estrus; n = 8), and NOMAS (cows that were inoculated but did not develop mastitis; n = 4). Ovulation rate was 100% for CON, NOMAS, and TRT-EST compared with 0% for TRT-NOEST cows. Size of the ovulatory follicle ("presumed" ovulatory follicle in TRT-NOEST cows) was similar for all groups. Frequency of LH pulses was decreased in TRT-NOEST compared with CON, TRT-EST, and NO-MAS. Estradiol-17beta increased over time in CON, NO-MAS, and TRT-EST cows, but did not increase in TRT-NOEST cows. Cows with clinical mastitis may exhibit estrus and ovulate normally or have disruptions in normal physiology including decreased LH pulsatility, absence of an LH surge and estrous behavior, suppressed estradiol-17beta, and failure to ovulate.


Machine learning based classification of yogurt aroma types with flavoromics.
Traditional sensory evaluation, relying on human assessors, is vulnerable to subjective error and lacks automation. Nonetheless, the complexity of human sensation makes it challenging to develop a computational method in place of human sensory evaluation. To tackle this challenge, this study constructed logistic regression classification models that could predict yogurt aroma types based on aroma-active compound concentrations with high classification accuracy (AUC ROC > 0.8). Furthermore, indicator compounds discovered from feature importance analysis of classification models led to the derivation of classification criteria of yogurt aroma types. Through constructing and analyzing machine learning models on yogurt aroma types, this study provides an automated pipeline to monitor sensory properties of yogurts.


Clarifying self-harm through evolutionary concept analysis.
Clarification of the concept self-harm is needed in order to enable research and theory development and facilitate the development and evaluation of medical interventions and nursing care for individuals who self-harm. This study presents such a conceptual analysis. Articles from 1997 to 2007 were sought from the Medline, PubMed, Cinahl, and PsychINFO search engines by entering the search words 'self-harm', 'self-harming', and 'psychiatric care'. 25 medicine and 23 nursing science articles were chosen for inclusion and analysed. Rodgers' evolutionary concept analysis process was used to delineate and clarify the concept's context, surrogate terms, antecedents, attributes, and consequences, as well as to determine implications for further research. Attributes of self-harm may include repetitive injury of mouth or exterior body, that is to say the infliction of physical pain to alleviate mental pain, and time spent self-harming. Antecedents may be gender, mental pain, substance abuse and relational problems. Consequences often include the need for medication and help with altering coping behaviour. Some self-harm patients met with negative attitudes from nurses. Individualized care and treatment is recommended. Accordingly, inter-professional collaboration and postgraduate education is needed in order to provide better care and treatment for self-harm patients. Furthermore, better understanding is needed to help enable health care personnel understand why individuals self-harm. The conceptual analysis presented in this study may be helpful as regards theory development within this still rather unexplored field.


Enhanced anticancer activity and endocytic mechanisms by polymeric nanocarriers of n-butylidenephthalide in leukemia cells.
The purpose of this study was to investigate the antitumor mechanisms of n-butylidenephthalide (BP) and to further examine the delivery efficacy of polycationic liposome containing PEI and polyethylene glycol complex (LPPC)-encapsulated BP in leukemia cells. MTS, flow cytometric and TUNEL assays were performed to assess cell viability and apoptosis. BP and BP/LPPC complex delivery efficiency was analyzed by full-wavelength fluorescent scanner and fluorescence microscope. The expressions of cell cycle- and apoptosis-related proteins were conducted by Western blotting. The results showed that BP inhibited leukemia cell growth by inducing cell cycle arrest and cell apoptosis. LPPC-encapsulated BP rapidly induced endocytic pathway activation, resulting in the internalization of BP into leukemia cells, causing cell apoptosis within 1 h. LPPC encapsulation enhanced the cytotoxic activity of BP and did not influence the effects of BP induction that suggested LPPC-encapsulated BP might be developed as anti-leukemia drugs in future.


Association between selective serotonin-reuptake inhibitor therapy and heart valve regurgitation.
The identification of an association between fenfluramines and valvular disease has raised the possibility of a similar association between another class of medications that increases local levels of serotonin, the selective serotonin-reuptake inhibitors (SSRIs). The objective of this study was to examine the association between heart valve regurgitation and treatment with SSRIs. We examined 5,437 consecutive patients who underwent echocardiography. Patients with a similar likelihood of SSRI treatment were identified by propensity models. The prevalence of regurgitation according to treatment was compared after adjusting for clinical characteristics associated with regurgitation. We also blindly reinterpreted a subset of 2,000 echocardiograms to identify characteristics associated with fenfluramine-associated valvular heart disease such as posterior mitral leaflet restriction. Among 5,437 consecutively hospitalized patients, we identified 292 who had taken SSRIs before admission. Patients taking SSRIs tended to be younger, female, Caucasian, unmarried, and more likely to have psychiatric illness and hypertension (p < or = 0.05). The overall prevalence of regurgitation meeting Food and Drug Administration criteria (at least moderate mitral regurgitation or mild aortic regurgitation) was 30%, with no significant difference in prevalence between those receiving SSRIs (26.7%) and controls (30.4%) (p = 0.19). The association remained negative when comparing SSRI-treated patients to controls with similar characteristics. Furthermore, the prevalence of features described in conjunction with fenfluramine exposure, such as posterior mitral leaflet restriction, was not higher in SSRI-treated patients. Among a large consecutive cohort of patients, the prevalence of mitral and aortic regurgitation in patients taking SSRIs was not different from that of controls, suggesting that SSRIs are not associated with valvular disease.


miRNA modulation of SOCS1 using an influenza A virus delivery system.
Difficulties associated with efficient delivery and targeting of miRNAs to cells is hampering the real world application of miRNA technology. This study utilized an influenza A-based delivery system to express miR-155 in order to knockdown SOCS1 mRNA. Using qPCR and dual luciferase technology we show that miR-155 delivery resulted in a significant increase in cellular miR-155 which facilitated a downregulation of SOCS1 gene expression and a functional increase in IL-6 and IFN-β cytokines.


Lower extremity functional tests and risk of injury in division iii collegiate athletes.
Functional tests have been used primarily to assess an athlete's fitness or readiness to return to sport. The purpose of this prospective cohort study was to determine the ability of the standing long jump (SLJ) test, the single-leg hop (SLH) for distance test, and the lower extremity functional test (LEFT) as preseason screening tools to identify collegiate athletes who may be at increased risk for a time-loss sports-related low back or lower extremity injury. A total of 193 Division III athletes from 15 university teams (110 females, age 19.1 ± 1.1 y; 83 males, age 19.5 ± 1.3 y) were tested prior to their sports seasons. Athletes performed the functional tests in the following sequence: SLJ, SLH, LEFT. The athletes were then prospectively followed during their sports season for occurrence of low back or LE injury. Female athletes who completed the LEFT in $118 s were 6 times more likely (OR=6.4, 95% CI: 1.3, 31.7) to sustain a thigh or knee injury. Male athletes who completed the LEFT in #100 s were more likely to experience a time-loss injury to the low back or LE (OR=3.2, 95% CI: 1.1, 9.5) or a foot or ankle injury (OR=6.7, 95% CI: 1.5, 29.7) than male athletes who completed the LEFT in 101 s or more. Female athletes with a greater than 10% side-to-side asymmetry between SLH distances had a 4-fold increase in foot or ankle injury (cut point: >10%; OR=4.4, 95% CI: 1.2, 15.4). Male athletes with SLH distances (either leg) at least 75% of their height had at least a 3-fold increase (OR=3.6, 95% CI: 1.2, 11.2 for the right LE; OR=3.6, 95% CI: 1.2, 11.2 for left LE) in low back or LE injury. The LEFT and the SLH tests appear useful in identifying Division III athletes at risk for a low back or lower extremity sports injury. Thus, these tests warrant further consideration as preparticipatory screening examination tools for sport injury in this population. The single-leg hop for distance and the lower extremity functional test, when administered to Division III athletes during the preseason, may help identify those at risk for a time-loss low back or lower extremity injury. 2.


Metabolite profiling reveals abiotic stress tolerance in Tn5 mutant of Pseudomonas putida.
Pseudomonas is an efficient plant growth-promoting rhizobacteria (PGPR); however, intolerance to drought and high temperature limit its application in agriculture as a bioinoculant. Transposon 5 (Tn5) mutagenesis was used to generate a stress tolerant mutant from a PGPR Pseudomonas putida NBRI1108 isolated from chickpea rhizosphere. A mutant NBRI1108T, selected after screening of nearly 10,000 transconjugants, exhibited significant tolerance towards high temperature and drought. Southern hybridization analysis of EcoRI and XhoI restricted genomic DNA of NBRI1108T confirmed that it had a single Tn5 insertion. The metabolic changes in the polar and non-polar extracts of NBRI1108 and NBRI1108T were examined using 1H, 31P nuclear magnetic resonance (NMR) spectroscopy and gas chromatography-mass spectrometry (GC-MS). Thirty six chemically diverse metabolites consisting of amino acids, fatty acids and phospholipids were identified and quantified. Insertion of Tn5 influenced amino acid and phospholipid metabolism and resulted in significantly higher concentration of aspartic acid, glutamic acid, glycinebetaine, glycerophosphatidylcholine (GPC) and putrescine in NBRI1108T as compared to that in NBRI1108. The concentration of glutamic acid, glycinebetaine and GPC increased by 34%, 95% and 100%, respectively in the NBRI1108T as compared to that in NBRI1108. High concentration of glycerophosphatidylethanolamine (GPE) and undetected GPC in NBRI1108 indicates that biosynthesis of GPE may have taken place via the methylation pathway of phospholipid biosynthesis. However, high GPC and low GPE concentration in NBRI1108T suggest that methylation pathway and phosphatidylcholine synthase (PCS) pathway of phospholipid biosynthesis are being followed in the NBRI1108T. Application of multivariate principal component analysis (PCA) on the quantified metabolites revealed clear variations in NBRI1108 and NBRI1108T in polar and non-polar metabolites. Identification of abiotic stress tolerant metabolites from the NBRI1108T suggest that Tn5 mutagenesis enhanced tolerance towards high temperature and drought. Tolerance to drought was further confirmed in greenhouse experiments with maize as host plant, where NBRI1108T showed relatively high biomass under drought conditions.


Gastrointestinal metabolism and transport of pesticidal carbamates.
Published evidence pertaining to gastrointestinal absorption and metabolism of pesticidal carbamates will be reviewed. The major body of evidence will come form investigations conducted by the author on N-methylcarbamate pesticides with rats; few investigations directly involving the gastrointestinal system have been reported by other research groups. Results derived from in vitro and isolated in vivo preparations show that pesticidal carbamates are absorbed and metabolized by gastrointestinal tissues. Numerous gastrointestinal metabolites have been separated; some have been identified and their behavior in the gut characterized; these data will be included in the review to the extent they have been published. The absorptive and metabolic capacities of the gastrointestinal system vary from one region to another. The author draws from evidence obtained from in vitro and in vivo studies and proposes the events that occur between carbamate ingestion and the appearance of the compound and/or its metabolites in the portal blood or mesenteric lymph. In this endeavor, the author acknowledges that superimposition of the metabolic precursory substrate-product sequence on the sequential physiologic steps from ingestion to defecation produces a complex situation which is difficult if not impossible to study meaningfully by investigation of one tissue component at a time (i.e., epithelial uptake, epithelial metabolism, mucosal to serosal transport, etc.).


Study of properties of phosphorylase kinase using hydrophobic chromatography.
The structure of nonactivated and activated forms of phosphorylase kinase has been investigated. The enzyme activation was achieved by phosphorylation with cAMP-dependent protein kinase as well as by incubation of the enzyme in an alkaline medium (pH 8.8). For structural comparison of the enzymic forms, hydrophobic chromatography on phenyl-Sepharose and polyacrylamide gel electrophoresis were used. It has been shown that the enzyme activation results in a release of a low molecular weight component (Mr 16 000). The properties of this component resemble those of calmodulin. Evidence for the formation of an unstable nonactivated phosphorylase kinase - calmodulin complex may be important for the correct understanding of the mechanism of enzyme activation.