Xenon/hypothermia neuroprotection regimes in spontaneously breathing neonatal rats after hypoxic-ischemic insult: the respiratory and sedative effects.
Hypothermia (HT) reduces neuronal injury after perinatal asphyxia. The anesthetic gas xenon (XE) may enhance this effect. We investigated the sedative and respiratory effects of variable XE concentrations at 37 degrees C normothermia (NT) or 32 degrees C HT after a hypoxic-ischemic (HI) insult to determine the concentration at which XE was a respiratory depressant in spontaneously breathing 7-day-old rat pups. (I) In three control groups, the effects of fasting at NT and HT were investigated. (II) Six groups were subjected to a HI insult (left carotid ligation then 90 min breathing 8% oxygen); three then breathed Air, 50%Xe or 70%Xe for 5 h at NT (NT(Air), NT(50%Xe), NT(70%Xe)), while three breathed identical mixtures during HT (HT(Air), HT(50%Xe), or HT(70%Xe)), in addition to a control group. Blood gases, glucose, and lactate were measured. Sedation (spontaneous movement/respiratory rate) was recorded. Blood chemistry data were successfully obtained from 70 pups. (I) Pups maintained normal blood gas, glucose, and lactate values after 9 h fasting at NT or HT. (II) After HI insult, in comparison with control and NT(Air) groups, 70%Xe at both NT and HT produced higher PCO2 and lower pH values while the HT(Air) and HT(50%Xe) groups only had lower pH values. The HT(70%Xe) combination produced the highest PCO2 and lowest pH values (56.8 mm Hg, 7.35, respectively) and the greatest sedative effect. After HI insult, 70%Xe at both NT and HT induced sedation, respiratory depression, CO2 retention, and a decrease in pH relative to air and control groups. The effects were largely avoided with 50%Xe.


An outbreak of Mayaro virus disease in Belterra, Brazil. III. Entomological and ecological studies.
Results in entomological and vertebrate host investigations made during dual outbreaks of Mayaro (MAY) and yellow fever (YF) viruses in Belterra, Pará, Brazil in 1978 are reported. Over 9,000 insects representing 26 species were assayed in 396 pools for the presence of arboviruses. Pools of Haemagogus janthinomys Dyar yielded the only isolates of either MAY or YF virus. The minimum field infection rate for nine isolates of MAY virus from Hg. janthinomys was 1:82, and for two isolates of YF virus was 1:368. Analysis of collection data showed Hg. janthinomys to be attracted to man as a blood source and present in all habitats sampled, although most abundant in the forest canopy. Twelve hundred bird sera and 584 mammal sera were tested by hemagglutination-inhibition (HI) tests for antibody to MAY virus. Highest MAY antibody prevalence rates were found among marmosets (Calithrix argentata, 32 positive of 119 tested, 27%). Mayaro virus was also isolated from the blood of a sylvan marmoset captured at the peak of the MAY virus outbreak. Experimental infection of marmosets with MAY virus confirmed that a substantial viremia follows infection with this virus. Marmosets were also found with HI antibody to YF virus (5/119, 4%). The results presented indicate that Hg. janthinomys was the principal vector of both MAY and YF viruses and that marmosets were the main amplifying hosts for MAY virus, and perhaps for YF virus as well.


[Status of cerebral and central hemodynamics in patients with dyscirculatory encephalopathy].
On the basis of radiocardiographic and radiocirculatory examination of the central and cerebral hemodynamics in 130 patients with dyscirculatory encephalopathy (DE) caused by essential hypertension and atherosclerosis, the authors have demonstrated that the development of DE is facilitated by disturbances of the central and peripheral hemodynamics. In turn, DE reduces the function of the autoregulatory mechanisms of the cerebral blood flow and leads to insufficiency of compensatory adaptive possibilities of the cardiovascular system. It is recommended that not only cerebral disorders but also the type (hyperkinetic, hypokinetic, eukinetic) of the blood circulation in these patients be taken into consideration in selecting the optimal method of treatment.


Plasma drug concentrations and clinical effects of a peripheral alpha-2-adrenoceptor antagonist, MK-467, in horses sedated with detomidine.
To investigate plasma drug concentrations and the effect of MK-467 (L-659'066) on sedation, heart rate and gut motility in horses sedated with intravenous (IV) detomidine. Experimental randomized blinded crossover study. Six healthy horses. Detomidine (10 μg kg(-1) IV) was administered alone (DET) and in combination with MK-467 (250 μg kg(-1) IV; DET + MK). The level of sedation and intestinal sounds were scored. Heart rate (HR) and central venous pressure (CVP) were measured. Blood was collected to determine plasma drug concentrations. Repeated measures anova was used for HR, CVP and intestinal sounds, and the Student's t-test for pairwise comparisons between treatments for the area under the time-sedation curve (AUCsed ) and pharmacokinetic parameters. Significance was set at p < 0.05. A significant reduction in HR was detected after DET, and HR was significantly higher after DET + MK than DET alone. No heart blocks were detected in any DET + MK treated horses. DET + MK attenuated the early increase in CVP detected after DET, but later the CVP decreased with both treatments. Detomidine-induced intestinal hypomotility was prevented by MK-467. AUCsed was significantly higher with DET than DET + MK, but maximal sedations scores did not differ significantly between treatments. MK-467 lowered the AUC of the plasma concentration of detomidine, and increased its volume of distribution and clearance. MK-467 prevented detomidine induced bradycardia and intestinal hypomotility. MK-467 did not affect the clinical quality of detomidine-induced sedation, but the duration of the effect was reduced, which may have been caused by the effects of MK-467 on the plasma concentration of detomidine. MK-467 may be useful clinically in the prevention of certain peripheral side effects of detomidine in horses.


Chemical Characterization of Capsule-Brewed Espresso Coffee Aroma from the Most Widespread Italian Brands by HS-SPME/GC-MS.
Coffee capsules market is on the rise as it allows access to a wide selection of coffee, differing in taste and brand. However, few data about the chemical characterization of the capsule-brewed coffee aroma are available. In this work, an untargeted approach using headspace solid-phase microextraction (HS-SPME) coupled to gas chromatography-mass spectrometry (GC-MS) and combined to chemometrics was performed to study and compare aroma profile from 65 capsule-brewed espresso coffees (ECs) commercialized by five of the most representative brands in Italy. Volatile profiles obtained from ECs were subjected to multivariate statistical analysis, which generally did not show a significant variability among coffees belonging to the same brand, except for those modified after the addition of specific flavor additives or aromatic substances (such as caramel, chocolate, etc.). Similarities may be related to the starting coffee brew or the processing method, which is likely the same for each individual brand. Additionally, partial least squares discriminant analysis (PLS-DA) showed that capsules from a specific brand contain the highest concentration of pyrazines, thus characterized by an intense and characteristic aroma, and a stronger note than those from the other brands. This study supports that the chemical analysis in conjunction with chemometric tools is a useful approach for assessing flavor quality, even if the need remains to identify volatile markers of high-quality beverages.


Deficient TACI expression on B lymphocytes of newborn mice leads to defective Ig secretion in response to BAFF or APRIL.
Capsular polysaccharides of encapsulated bacteria do not induce immune response in newborns and the mechanism for this unresponsiveness is not clear. In adults, transmembrane activator and calcium-modulator and cyclophilin [corrected] ligand interactor (TACI) is a TNFR family member molecule with a pivotal role in Ab responses against polysaccharide vaccines. We investigated the expression and the functions of the TNF family cytokines, B cell-activating factor of the TNF family (BAFF) and a proliferation-inducing ligand (APRIL), and their receptors in newborn mice and found that TACI expression on B lymphocytes was dramatically reduced (p < 0.0001) in newborns as compared with adults. More importantly, TACI ligands BAFF or APRIL were unable to induce IgA/IgG/IgM secretion from newborn B lymphocytes. Additionally, TACI expression seems to be important in plasma cell development. Indeed, in contrast to adults, stimulation of newborn B lymphocytes with BAFF or APRIL did not result in up-regulation of CD138 expression. In vitro or in vivo exposure of newborn B lymphocytes to oligodeoxynucleotides (CpG ODN) led to up-regulation of TACI expression on newly formed, follicular, and marginal zone as well as B1 B lymphocyte populations, and rendered them responsive to BAFF- or APRIL-mediated CD138 expression and IgA/IgG secretion. Finally, immunization of newborn BALB/c mice but not TACI knockout mice with CpG ODN containing (4-hydroxy-3-nitrophenyl)acetyl-Ficoll led to development of IgG Abs against (4-hydroxy-3-nitrophenyl)acetyl. These findings demonstrate that low TACI expression may be a critical factor that determines the susceptibility of newborns to infections with encapsulated bacteria and the impaired immunogenicity of polysaccharide vaccines. Finally, CpG ODNs may correct deficient newborn response to polysaccharide vaccines by up-regulating TACI.


The number of vertebrae in the Southern African Negro, the American Negro and the Bushman (San).
The numbers of precoccygeal vertebrae (PCV) are compared in Southern African Negroes, Bushman (San) and American Negroes. The S.A. Negro and San males have an exceptionally high frequency of an increased total PCV number whilst the frequencies are significantly lower in a combined Caucasoid sample and the Mongoloid samples. The American Negro male, too, has a high frequency but not as high as in S.A. Negroes. The high frequency of increased PCV number in San, S.A. Negroes and American Negroes is in keeping with the view that the Khoisan peoples (here represented by the San), the Southern African Negroes and the African ancestors of American Blacks sprang from a common proto-negriform stock. Males have a significantly greater frequency of increased PCV to 30 or 31, whereas females are more likely to have a decreased number of 28 PCV. This sex difference is present in all the populations studied.


Expression of anatomical leaf traits in homoploid hybrids between deciduous and evergreen species of Vaccinium.
We investigated the anatomical expression of leaf traits in hybrids between evergreen Vaccinium vitis-idaea and deciduous V. myrtillus. We compared parents from four populations with their respective F1 hybrids and tested whether (i) transgression can be the source of novel anatomical traits in hybrids; (ii) expression of transgressive traits is more probable for traits with similar values in parents and intermediate for more distinct values, as predicted by theory; and (iii) independent origin of hybrids leads to identical trait expression profiles among populations. We found that anatomical leaf traits can be divided into four categories based on their similarity to parents: intermediate, parental-like, transgressive and non-significant. Contrary to the common view, parental-like trait values were equally important in shaping the hybrid profile, as were intermediate traits. Transgression was revealed in 17/144 cases and concerned mainly cell and tissue sizes. As predicted by theory, we observed transgressive segregation more often when there was little phenotypic divergence, but intermediate values when parental traits were differentiated. It is likely that cell and tissue sizes are phylogenetically more conserved due to stabilising selection, whereas traits such as leaf thickness and volume fraction of the intercellular spaces, showing a consistent intermediate pattern across populations, are more susceptible to directional selection. Hybrid populations showed little similarity in expression profile, with only three traits identically expressed across all populations. Thus local adaptation of parental species and specific genetic background may be of importance.


The influence of pCO2 and temperature on gene expression of carbon and nitrogen pathways in Trichodesmium IMS101.
Growth, protein amount, and activity levels of metabolic pathways in Trichodesmium are influenced by environmental changes such as elevated pCO(2) and temperature. This study examines changes in the expression of essential metabolic genes in Trichodesmium grown under a matrix of pCO(2) (400 and 900 µatm) and temperature (25 and 31°C). Using RT-qPCR, we studied 21 genes related to four metabolic functional groups: CO(2) concentrating mechanism (bicA1, bicA2, ccmM, ccmK2, ccmK3, ndhF4, ndhD4, ndhL, chpX), energy metabolism (atpB, sod, prx, glcD), nitrogen metabolism (glnA, hetR, nifH), and inorganic carbon fixation and photosynthesis (rbcL, rca, psaB, psaC, psbA). nifH and most photosynthetic genes exhibited relatively high abundance and their expression was influenced by both environmental parameters. A two to three orders of magnitude increase was observed for glnA and hetR only when both pCO(2) and temperature were elevated. CO(2) concentrating mechanism genes were not affected by pCO(2) and temperature and their expression levels were markedly lower than that of the nitrogen metabolism and photosynthetic genes. Many of the CO(2) concentrating mechanism genes were co-expressed throughout the day. Our results demonstrate that in Trichodesmium, CO(2) concentrating mechanism genes are constitutively expressed. Co-expression of genes from different functional groups were frequently observed during the first half of the photoperiod when oxygenic photosynthesis and N(2) fixation take place, pointing at the tight and complex regulation of gene expression in Trichodesmium. Here we provide new data linking environmental changes of pCO(2) and temperature to gene expression in Trichodesmium. Although gene expression indicates an active metabolic pathway, there is often an uncoupling between transcription and enzyme activity, such that transcript level cannot usually be directly extrapolated to metabolic activity.


A novel patient-oriented numerical procedure for glaucoma drainage devices.
The present work analyses the performance of four glaucoma drainage devices, by means of a novel patient-oriented numerical procedure. The procedure is based on the three-dimensional geometry reconstruction from the stacks of tomographic images of a human eye, at different angles, on meshing and on thermo-fluid dynamics modelling activities, carried out on the reconstructed computational domain. The current three-dimensional eye model considers anterior chamber (AC), trabecular meshwork, Schlemm's canal, and collector channels, making use of generalised porous medium approach for modelling ocular porous tissue and cavities. The intraocular pressure (IOP) management inside AC of human eye is analysed, by comparing the results obtained for four drainage devices implanted in a human eye for glaucoma treatment, ie, ExPRESS shunt, iStent inject, SOLX gold micro shunt, and the novel silicon shunt device. The numerical results allow predicting the effects of the installation of these implants on human eyes, in terms of IOP decrease, aqueous humour velocity, pressure, friction coefficient, and local Nusselt number, pointing out the clear distinction between pre-operative and post-operative eye conditions for different glaucoma surgical techniques.


Targeted gene delivery to human osteosarcoma cells with magnetic cationic liposomes under a magnetic field.
Gene delivery using cationic liposomes results in relatively low transfection, especially under in vivo conditions. This system, however, can overcome some of the problems associated with viral delivery systems. The present study was carried out in order to improve the transfection efficiency of cationic liposomes by preparing magnetic cationic liposomes (MCL). Small MCL approximately 40 nm in diameter and incorporating one or two magnetite particles were prepared with phosphatidylethanolamine and 3beta-[N-(N', N'-dimethylaminoethane)-carbamoyl] cholesterol. The efficiency of MCL in gene delivery was evaluated by using plasmid DNA containing a luciferase reporter gene and human osteosarcoma Saos-2 cells. Without a magnetic field, maximum luciferase activity was observed when DNA was mixed with MCL at a 1:5 ratio and incubated with cells for 6 h. Under a magnetic field, maximum luciferase activity was achieved by 30-min magnetic induction. This improvement in transfection efficiency by magnetic induction was approximately 3.5-fold. The feasibility of this active transgenic system was further shown by measuring apoptosis rates after transfection of the p53 gene to Saos-2 cells. Apoptosis rates increased to 18.9% from 2.4% by magnetic induction. In conclusion, a gene delivery system including MCL and magnetic induction was found to achieve rapid and enhanced gene delivery in vitro. Such a gene delivery system may be applicable under in vivo conditions, and is expected to offer numerous clinical advantages.


Discovering dad: paternal roles, responsibilities, and support needs as defined by fathers of children with complex cardiac conditions perioperatively.
Understanding perceptions of family caregivers' roles and responsibilities regarding their child with complex cardiac needs has potential to help care teams better support parents. Paternal experience has been under-explored in pediatric cardiac cohorts. Ten fathers of children undergoing cardiac surgery completed quantitative surveys on their knowledge needs and preferred format of communication. In face-to-face recorded interviews, they responded to open-ended questions about the definition of being a good father to a child with a complex cardiac condition, perceived paternal responsibilities, personal growth as a parent to a child with a complex heart condition, support needs, and recommendations to medical staff for paternal inclusion. Semantic content analysis was utilised. The study reports strictly followed COnsolidated criteria for REporting Qualitative research guidelines. The fathers reported high preference for knowledge about the child's heart condition, communication about the treatment plan, and desire for inclusion in the care of their child. Paternal role was defined thematically as: providing a supportive presence, being there, offering bonded insight, serving as strong provider, and acting as an informed advocate. The fathers revealed that their responsibilities sometimes conflicted as they strove to serve as an emotional and economic stabiliser for their family, while also wanting to be foundationally present for their child perioperatively. This study provides insight into paternal experience and strategies for paternal inclusion. This summary of the self-defined experience of the fathers of pediatric cardiac patients offers constructive and specific advice for medical teams.


Excystation and culture of Giardia spp. from human source.
In this paper, we report the hatching of Giardia spp. trophozoites from purified cysts harvested in Sheather's solution from human feces. Isolated cysts were induced by acidic Hank's Balanced Salt Solution (HBSS), incubated at 37 degrees C for 30 minutes, concentrated by centrifugation and washed with pH 7.5 HBSS. The cysts were placed in 8 ml. culture medium (Pancreatic digest of casein and yeast extracts) supplemented with human serum and bile in borosilicate glass tubes with screw caps. Trophozoite excystation was scanned in a Commandon chamber filled with the suspension, sealed with a cover glass and vaseline. The trophozoite emergence was recorded with phase contrast in a photomicroscope. The excystation began within the first 3 minutes, after inoculation into medium. The trophozoites emerged trough a small hole at one end of the cyst and simultaneously began cellular division, the whole process required 5-30 minutes. The trophozoites attached themselves to the walls of the culture tube, permitting the replacement and removal of contaminants without dislodging them from the tube walls. The isolates MM-INP and MG-INP have been maintained at 37 degrees C for more than six months.


Impact of traumatic dental injuries on quality of life in preschoolers and schoolchildren: A systematic review and meta-analysis.
Traumatic dental injuries (TDIs) in childhood and adolescence are a potential public health problem given their prevalence and consequences. The aim of this study was to assess the impact of TDIs on the oral health-related quality of life (OHRQoL) of preschoolers and schoolchildren, by synthesizing the available evidence. A systematic search was conducted in MEDLINE, EMBASE, Cochrane, ScieLo and Lilacs databases since January 1966 until March 2016. The included studies compared OHRQoL between groups with and without TDIs, using validated instruments. The selection process and data extraction were carried out by two researchers independently. A third reviewer resolved discrepancies. Methodological quality was assessed with the Effective Public Health Practice Project's Quality Assessment Tool. Meta-analyses were performed using random effect models, separately for preschoolers and schoolchildren. Of 213 identified articles, 26 studies (involving a total of 4582 patients and 13 601 controls between the ages of 1 and 15 years) met the inclusion criteria. Most of the studies had been published in the last 5 years, and their methodological quality was judged to be moderate. The TDIs group had a significantly higher chance of reporting any impact on OHRQoL than controls for both preschoolers (OR = 1.44; 95% confidence interval [CI]: 1.28-1.63; I2 = 0%) and schoolchildren (OR = 1.31; 95% CI: 1.04-1.66; I2 = 70%). In preschoolers, the OR for OHRQoL impact for complicated vs uncomplicated TDIs was 1.53 (95% CI: 1.04-2.26; I2 = 0%). The social domain was the most affected one in schoolchildren (standard mean difference = 0.34; 95% CI: 0.13, 0.55; I2 = 68%). Traumatic dental injuries have a negative impact on OHRQoL of both preschoolers and schoolchildren. Outcome standardization to measure OHRQoL impact, such as mean score differences and cut-off points, is needed. Prospective cohort studies are recommended to confirm these findings and to understand how TDIs' impact changes with time.


Evaluation of postoperative discomfort after impacted mandibular third molar surgery using three different types of flap.
The surgical extraction of an impacted third molar involves a wide range of consequences such as trismus, swelling, and pain, as well as more significant complications, temporary or permanent, that can manifest altered sensitivity of the tongue or lips. The purpose of this prospective study was to evaluate the effects of three different flaps on postoperative discomfort considering trismus, edema, and pain, after the extraction of impacted third molars. The data derived from the analysis of the surgical trials performed at the Oral Surgery Unit, Department of Surgical, Reconstructive and Diagnostic Sciences, IRCCS Policlinico, University of Milan, directed by Professor F. Santoro, MD. This study, developed over 2 years, involved 238 patients for a total of 238 extractions of impacted mandibular third molars. The 238 surgeries were performed on 114 men and 124 women: 54 avulsions were performed with the elevation of an envelope flap (Group 1), 48 avulsions through the elevation of a triangular flap (Group 2), and the remaining 136 avulsions were performed using a trapezoidal flap (Group 3). Trismus was significantly reduced (P < .05) in patients treated with envelope flap, as was the swelling perceived by the patient (P < .05). Pain was closely related to the elevation of a mucoperiosteal flap and osteotomy. Our study does not reveal statistically significant differences between the three types of flap used; however, the number of analgesic tablets taken was lower in cases of elevation of a less traumatic flap (envelope and triangular flaps). The data collected in this study indicate the envelope flap as the most suitable for the reduction of the expression of postoperative complications such as swelling and trismus.


The quick and the dead: "managing" inpatient care in British Columbia hospitals, 1969-1995/96.
To examine changes in hospital use in British Columbia during a decade of capacity reductions. The data used are all separation records for British Columbia hospitals for the years 1969, 1978, 1985/86, 1993/94, and 1995/96. Separation records include acute care, rehabilitation, extended care, and surgical day care hospital encounters in British Columbia that were concluded during the years of interest. Analyses were based on per capita use of services for five-year age groups of the population to ages 90+; the emphasis was on looking at changes in the use of specific types of hospital services over the 26 years of study, with a particular focus on the most recent decade. Data were extracted from hospital separations files owned by the British Columbia Ministry of Health and housed at the Centre for Health Services and Policy Research. All separation records for the years of interest were included in the study. Acute care use continued to fall over the last decade. The rate of decline increased during the last time period of study and affected seniors to the same degree as younger patients. At the same time, use of extended care decreased, compared to steady increases in earlier years. The result was that by 1995/96 nearly 40 percent of inpatient days were used by people who died in hospital, compared to 9 percent in 1969. These people, however, still represent a small proportion of separations. The "bed blocker" problem common to many hospital systems appears to have been largely alleviated in British Columbia over the decade 1985-95. The concurrent decrease in extended care use, however, makes it difficult to say where and how these people are now being cared for. Care for the dying has become a bigger issue for hospitals, but whether this is because of heroic interventions at the end of life is not clear. A "top-down," capacity-driven management approach to hospital use in British Columbia has produced effects that may seem familiar to those involved in more "bottom-up" managed care approaches in the United States.


A vessel noise budget for Admiralty Inlet, Puget Sound, Washington (USA).
One calendar year of Automatic Identification System (AIS) ship-traffic data was paired with hydrophone recordings to assess ambient noise in northern Admiralty Inlet, Puget Sound, WA (USA) and to quantify the contribution of vessel traffic. The study region included inland waters of the Salish Sea within a 20 km radius of the hydrophone deployment site. Spectra and hourly, daily, and monthly ambient noise statistics for unweighted broadband (0.02-30 kHz) and marine mammal, or M-weighted, sound pressure levels showed variability driven largely by vessel traffic. Over the calendar year, 1363 unique AIS transmitting vessels were recorded, with at least one AIS transmitting vessel present in the study area 90% of the time. A vessel noise budget was calculated for all vessels equipped with AIS transponders. Cargo ships were the largest contributor to the vessel noise budget, followed by tugs and passenger vessels. A simple model to predict received levels at the site based on an incoherent summation of noise from different vessels resulted in a cumulative probability density function of broadband sound pressure levels that shows good agreement with 85% of the temporal data.


Higher dementia incidence in older adults with type 2 diabetes and large reduction in HbA1c.
although type 2 diabetes increases risk of dementia by 2-fold, whether optimizing glycemic level in late life can reduce risk of dementia remains uncertain. We examined if achieving the glycemic goal recommended by the American Diabetes Association (ADA) within a year was associated with lower risk of dementia in 6 years. in this population-based observational study, we examined 2246 community-living dementia-free Chinese older adults with type 2 diabetes who attended the Elderly Health Centres in Hong Kong at baseline and followed their HbA1c level and cognitive status for 6 years. In line with the ADA recommendation, we defined the glycemic goal as HbA1c < 7.5%. The study outcome was incident dementia in 6 years, diagnosed according to the 10th revision of the International Statistical Classification of Diseases and Related Health Problems (ICD-10) or Clinical Dementia Rating of 1-3. those with HbA1c ≥ 7.5% at baseline and HbA1c < 7.5% in 1 year were associated with higher rather than lower incidence of dementia, independent of severe hypoglycemia, glycemic variability and other health factors. Sensitivity analyses showed that a relative reduction of ≥10%, but not 5-10%, in HbA1c within a year was associated with higher incidence of dementia in those with high (≥8%) and moderate (6.5-7.9%) HbA1c at baseline. a large reduction in HbA1c could be a potential predictor and possibly a risk factor for dementia in older adults with type 2 diabetes. Our findings suggest that optimizing or intensifying glycemic control in this population requires caution.


Epstein-Barr virus leader protein enhances EBNA-2-mediated transactivation of latent membrane protein 1 expression: a role for the W1W2 repeat domain.
The Epstein-Barr virus (EBV)-encoded leader protein EBNA-LP is made up of several 66-amino-acid repeats (the W1W2 domains) linked to a unique 45-amino-acid C-terminal sequence (the Y1Y2 domain). This protein is highly expressed along with a second nuclear antigen, EBNA-2, during the initial stages of virus-induced B-cell transformation. While EBNA-2's essential role in transformation as a transcriptional activatory is well documented, very little is known about EBNA-LP function except that recombinant viruses lacking the EBNA-LP Y1Y2 exons show reduced, but still detectable, transforming ability. This was taken as evidence that EBNA-LP plays an auxiliary role but is not essential for transformation. A recent study showed that EBNA-LP could cooperate with EBNA-2 in activating cyclin D2 transcription in resting B cells (A.J. Sinclair, L Palmero, G. Peters, and P.J. Farrell, EMBO J. 13:3321-3328, 1994). Here we report that EBNA-LP can also cooperate with EBNA-2 in up-regulating expression of the major EBV effector protein of B-cell transformation, latent membrane protein 1 (LMP1). In transient-transfection assays, EBNA-LP enhanced the level of EBNA-2-induced LMP1 expression by 5- to 10-fold in one Latency I Burkitt's lymphoma cell line, Eli-BL, and was absolutely required, along with EBNA-2, to induce LMP1 in a second line, Akata-BL. These changes in LMP1 protein expression appeared to be reflected at the transcriptional level. A study of EBNA-LP mutants showed that this cooperative function mapped to the W1W2 repeat domain rather than to Y1Y2. Because a Y1Y2-deleted form of EBNA-LP may therefore retain some aspects of wild-type function, the original data from virus recombinants leave open the possibility that EBNA-LP is actually an essential transforming gene.


The Bench-to-Bedside Story of IL-17 and the Therapeutic Efficacy of its Targeting in Spondyloarthritis.
TNF-blocking biologics have revolutionized the care of patients with spondyloarthritis, a group of clinically overlapping conditions that includes ankylosing spondylitis and psoriatic arthritis. However, incomplete response rates speak to the need for alternative therapeutic approaches. Over the last decade, animal models, genetics, and translational studies have implicated the excessive production of a pro-inflammatory cytokine interleukin-17 (IL-17) along with another IL-17-promoting cytokine IL-23 in the pathogenesis of spondyloarthritis. Genome-wide studies identified disease associations with multiple genes regulating IL-23/IL-17 immune pathway activity. Direct examination of the patient blood and tissues revealed excessive IL-17 and IL-23 production by diverse cell types. Murine models both underscored the sufficiency of excess IL-23 in driving disease phenotype and predicted utility in IL-23/IL-17 pathway blockade. However, the clinical efficacy of agents such as secukinumab and ustekinumab, which block IL-17 and IL-23/IL-12 respectively, provided exciting proof of concept.


Obesity education for front-line healthcare providers.
To assess the effect of an interprofessional educational activity on professional skills, attitudes, and perceived challenges toward obesity management among front-line healthcare providers. A one-day interprofessional obesity education activity was organized for healthcare providers across various disciplines. All participants were invited to complete an anonymous survey pre- and post-event, and at six-month post-event. The survey was created based on a comprehensive list of perceived skills, professional attitudes and challenges toward obesity intervention compiled from existing literature. Sixty-seven healthcare providers completed the survey pre- and post-event. Participants reported increases in professional skills such as their ability to assess weight (p = 0.04), to address weight management issues (p < 0.001), to teach/motivate patients toward physical activity (p < 0.001) and healthy eating practices (p = 0.001), to use behavior modification techniques (p < 0.001), and to deal with family issues (p < 0.001). Professional attitudes: practitioners felt more educated/competent in obesity management (P < 0.001), learned where to refer patients (p < 0.001), were more comfortable in discussing obesity in managing obesity (p < 0.001), were less likely to avoid the topic (p = 0.004) and felt less frustrated with the low success rate (p = 0.030). Enhancement in professional attitudes remained 6 months after the event. Improvements were mainly associated with male gender, younger age, fewer years of professional practice and healthcare professionals other than physicians. No statistically significant changes in perceived challenges were found after the educational event. Results of this study showed that this interprofessional learning activity contributed to the improvement of professional skills and attitudes of front-line healthcare providers caring for those who are obese or at risk for obesity. The positive results of this interprofessional learning activity aligns with the training needs identified by healthcare practitioners in previous studies, and suggest that this design and content could be used to guide future educational programming in the care of obese people.


A reassessment of P2X7 receptor inhibition as a neuroprotective strategy in rat models of contusion injury.
These experiments were completed as part of an NIH "Facilities of Research Excellence in Spinal Cord Injury" contract to support independent replication of published studies that could be considered for eventual clinical testing. Recent studies have reported that selective inhibition of the P2X7 receptor improves both the functional and histopathological consequences of a contusive spinal cord injury (SCI) in rats. We repeated two published studies reporting the beneficial effects of pyridoxal-5'-phosphate-6-azophenyl-2'-4'-disulphonic acid (PPADS) or Brilliant blue G (BBG) treatment after SCI (Wang et al., 2004 and Peng et al., 2009). Mild thoracic SCI was first produced in Experiment 1 by means of the MASCIS impactor at T10 (height 6.25 mm, weight 10 g) followed by intraspinal administration of a P2X7 antagonist (2 μl/10 mM) after injury. Treatment with PPADS or another highly selective P2X7R antagonist Brilliant Blue G (BBG) (2 μl/02 mM) did not improve locomotive (BBB rating scale) over a 7 week period compared to vehicle treated rats. Also, secondary histopathological changes in terms of overall lesion and cavity volume were not significantly different between the PPADS, BBG, and vehicle treated animals. In the second experiment, the systemic administration of BBG (10 or 50 mg/kg, iv) 15 min, 24 and 72 h after moderate (12.5 mm) SCI failed to significantly improve motor recovery or histopathological outcome over the 6 week observational period. Although we cannot conclude that there will be no long-term beneficial effects in other spinal cord injury models using selective P2X7 receptor antagonists at different doses or treatment durations, we caution researchers that this potentially exciting therapy requires further preclinical investigations before the implementation of clinical trials targeting severe SCI patients.


Iohexol versus metrizamide for cervical myelography: double-blind trial.
Cervical myelography was performed by lateral C1-C2 puncture in 60 patients. Thirty patients received iohexol (an investigational aqueous contrast agent) and 30 received metrizamide in order to provide a prospective, randomized, double-blind trial comparing these contrast media. The two media produced radiographs of equal quality. Contrast-related morbidity was suffered by four patients (13%) in the iohexol group and by 11 patients (37%) in the metrizamide group. These features indicate that iohexol is superior to metrizamide as a contrast agent for cervical myelography.


Two-Dimensional NMR Lineshape Analysis.
NMR titration experiments are a rich source of structural, mechanistic, thermodynamic and kinetic information on biomolecular interactions, which can be extracted through the quantitative analysis of resonance lineshapes. However, applications of such analyses are frequently limited by peak overlap inherent to complex biomolecular systems. Moreover, systematic errors may arise due to the analysis of two-dimensional data using theoretical frameworks developed for one-dimensional experiments. Here we introduce a more accurate and convenient method for the analysis of such data, based on the direct quantum mechanical simulation and fitting of entire two-dimensional experiments, which we implement in a new software tool, TITAN (TITration ANalysis). We expect the approach, which we demonstrate for a variety of protein-protein and protein-ligand interactions, to be particularly useful in providing information on multi-step or multi-component interactions.


[Radiation-induced changes in the genome of pine seeds, forming under conditions of chronic irradiation in the Chernobyl exclusion zone].
Comparative study of long-term existing genome damages in the chronically irradiated pine seeds from different parts of Chernobyl zone have been performed by a pulsed field gel electrophoresis and hydroxyapatite chromatography. Some differences were established for DNA degradation kinetic from protoplasts with various accumulated doses of irradiation. Results obtained could testify the existence of long-living changes in the experimental plants genome.


PET Neuroimaging: Insights on Dystonia and Tourette Syndrome and Potential Applications.
Primary dystonia (pD) is a movement disorder characterized by sustained or intermittent muscle contractions causing abnormal, often repetitive, movements, postures, or both. Gilles de la Tourette syndrome (GTS) is a childhood-onset neuropsychiatric developmental disorder characterized by motor and phonic tics, which could progress to behavioral changes. GTS and obsessive-compulsive disorders are often seen in comorbidity, also suggesting that a possible overlap in the pathophysiological bases of these two conditions. PET techniques are of considerable value in detecting functional and molecular abnormalities in vivo, according to the adopted radioligands. For example, PET is the unique technique that allows in vivo investigation of neurotransmitter systems, providing evidence of changes in GTS or pD. For example, presynaptic and post-synaptic dopaminergic studies with PET have shown alterations compatible with dysfunction or loss of D2-receptors bearing neurons, increased synaptic dopamine levels, or both. Measures of cerebral glucose metabolism with (18)F-fluorodeoxyglucose PET ((18)F-FDG PET) are very sensitive in showing brain functional alterations as well. (18)F-FDG PET data have shown metabolic changes within the cortico-striato-pallido-thalamo-cortical and cerebello-thalamo-cortical networks, revealing possible involvement of brain circuits not limited to basal ganglia in pD and GTS. The aim of this work is to overview PET consistent neuroimaging literature on pD and GTS that has provided functional and molecular knowledge of the underlying neural dysfunction. Furthermore, we suggest potential applications of these techniques in monitoring treatments.


Complications of aortic aneurysm surgery: prevention and treatment.
The most common complications associated with resection of aneurysms of the thoracic and abdominal aorta are: hemorrhage, acute renal failure, ischemic colitis, distal emboli, graft thrombosis, infection, pseudoaneurysm formation, aorto-caval and aorto-enteric fistulae, neurologic deficits, ureteral obstruction, sexual dysfunction, chylous ascites and perigraft seroma. Complications can be immediate or late sequelae, insiduous or life threatening. Their prevention and treatment, should they occur, will be discussed.


Use of IGK gene rearrangement analysis for clonality assessment of lymphoid malignancies: a single center experience.
Diagnosis of B-non Hodgkin lymphomas (NHLs) is based on clinical, morphological and immunohistochemi-cal features. However, in up to 10-15% of cases, analysis of immunoglobulin heavy (IGH) or light (IGK/IGL) chains genes is required to discriminate between malignant and reactive lymphoid proliferations. In this study, we evaluated the feasibility and efficiency of IGK analysis in the routine diagnostic of B-cell lymphoproliferative disorders (B-LD) when applied to formalin-fixed paraffin-embedded (FFPE) tissues. Clonality patterns were studied in 59 B-LD using the BIOMED-2 protocol for IGK assays, after failure of the IGH assay. PCR products were evaluated by both heterodu-plex and GeneScan analysis. IGK analysis was technically successful in all cases. Overall, it supported the histopa-thological suspicion in 52/59 cases (88%), the sensitivity and specificity being 83% and 80%, respectively. Further, positive and negative predictive values were 95% and 50%, respectively. Interestingly, among various lymphoma subtypes, marginal zone lymphoma and follicular lymphoma most frequently required IGK analysis. In conclusion, IGK study according to the BIOMED-2 protocol resulted feasible and extremely useful in supporting challenging diagnosis of B-LD even if applied on FFPE samples. Accordingly, when NHL is suspected, negative results at IGH analysis should not be considered as conclusive and further investigation of IGK is appropriate.


Carrageenan-induced acute inflammation in the mouse air pouch synovial model. Role of tumour necrosis factor.
We used the mouse air pouch model of inflammation to study the interaction between cytokines, prostaglandin E(2) (PGE(2)) and cell migration during the various phases of acute local inflammation induced by carrageenan. In serum, the levels of interleukin 1 (IL-1), interleukin 6 (IL-6), tumour necrosis factor (TNF), serum amiloid-A (SAA) and Fe(++) were never different from controls, indicating that no systemic inflammatory changes were induced. Locally the exudate volume and the number of leukocytes recruited into the pouch increased progressively until 7 days after carrageenan. The same was true for PGE(2) production. We could not measure IL-1 but the production of IL-6 and TNF reached a maximum after 5-24 h then quickly decreased. Anti-TNF antibodies inhibited cell migration by 50% 24 h after treatment. Pretreatment with interleukin 10 (IL-10) inhibited TNF production almost completely and cell migration by 60%. Carrageenan-induced inflammation was modulated by anti-inflammatory drugs. Pretreatment with dexamethasone (DEX) or indomethacin (INDO) inhibited cell migration and reduced the concentration of TNF in the exudate. Production of PGE(2) or vascular permeability did not correlate with the number of cells in the pouch. Local TNF seems to play an important role in this model, particularly for leukocyte migration in the first phase of the inflammatory process. In conclusion, the air pouch seems to be a good model for studying the regulation of the early events of local inflammation, particularly the role of cytokines and cell migration.


Cytokine response during hyperoxia: sequential production of pulmonary tumor necrosis factor and interleukin-6 in neonatal rats.
Exposure of newborn animals to high concentrations of oxygen leads to diffuse alveolar damage similar to that seen in bronchopulmonary dysplasia in human infants. Therefore, neonatal rats are a suitable practical model of hyperoxic lung damage in human infants. To determine the involvement of tumor necrosis factor-alpha and interleukin-6 in lung injury in neonatal rats exposed to 100% O2 concentration. A randomized controlled study was designed in which litters of term Sprague-Dawley rat pups were assigned to experimental or control groups. The pups in the experimental group were placed in 100% O2 from birth for 9 days, while the control pups were placed in room air. Twelve to 15 pups from each group were sacrificed on day 1, 3, 6, 9 and 13 after birth for bronchoalveolar lavage collection and lung histologic study. The bronchoalveolar lavage fluid was assayed for TNF alpha and IL-6. Newborn rats exposed to 100% O2 for the first 9 days of life showed severe pulmonary edema and hypercellularity on days 1 and 3, which then improved to nearly complete resolution on days 6 and 9. Pulmonary TNF alpha was produced early on O2 exposure (day 3) and pulmonary IL-6 later (days 6 and 9). Hyperoxia induces sequential production of pulmonary TNF alpha and IL-6, which corresponds to the severity of the pathological findings and the known inflammatory and anti-inflammatory role of these cytokines.


Memory suppressor T cells in latent Japanese encephalitis virus infection.
The generation of secondary suppressor T (Ts) cells has been studied during latent Japanese encephalitis virus (JEV) infection of mice. The mice infected with JEV 27 weeks earlier, on challenge with the homologous virus, showed accelerated generation of secondary Ts cells; these appeared on Day 6, with peak activity on Day 8, and lasted for 27 days. The secondary Ts cells were Thy1.2+, Ly1-2+, antigen-specific, and acted in a dose-dependent manner. The secondary Ts cells could also be generated by reactivation of the JEV in latently infected mice. The findings thus show the presence of memory suppressor T cells in mice latently infected with JEV that can be stimulated to produce secondary Ts cells by exogenous or endogenous virus challenge. This phenomenon could help to persistence of the virus.


Direct immunoelectron microscopy of transmissible gastroenteritis virus with immunoglobulins G and A and guinea pig complement.
Porcine colostral immunoglobulin (Ig)G and IgA, isolated from transmissible gastroenteritis virus-infected sows, were compared by direct immunoelectron microscopy. It was estimated, using antibodies with a less than a twofold difference in virus-neutralizing activity, that IgG was 500 times more efficient than was IgA for coating transmissible gastroenteritis virions. Guinea pig complement enhanced the antibody coating with IgG, but did not increase virus-neutralizing activity of IgG or IgA.


Lactate oxidation and sodium reabsorption by dog kidney in vivo.
Lactate uptake (Qlact) and oxidation (QCO2lact), oxygen consumption (QO2) and net tubular Na+ reabsorption (TNa), were estimated in pentobarbital-anesthetized dogs under control conditions and following unilateral intrarenal injection of ouabain or intravenous infusion of acetazolamide, ethacrynic acid, or furosemide. QCO2lact accounted for approximately half of simultaneous Qlact and for about one-third of QO2 in control periods. Ouabain injection resulted in significant decreases in several functions of the injected kidney: TNa, 46%; TNa/FNa, 36%; QO2, 40%; Qlact, 59%; and QCO2lact, 70%. Acetazolamide infusion decreased TNa, 33%; TNa/FNa, 12%; QO2, 10%; and QCO2lact, 38%; but did not change Qlact. Ethacrynic acid diminished TNa, 60%; TNa/FNa, 36%; QO2, 45%; Qlact, 31%; and QCO2lact, 73%. Furosemide lowered TNa, 37%; TNa/FNa, 28%; QO2, 25%; and Qlact, 48%; but did not change QCO2lact, 2%. Results indicate that decarboxylation is a major pathway of renal lactate metabolism, that lactate oxidation is a substantial source of aerobic energy for the kidney, and that QCO2lact is probably functionally related to sodium reabsorption. This relationship appears to be closer for a fraction of TNa associated with ouabain- and ethacrynic acid-sensitive mechanisms.


Skin-specific expression of ictacalcin, a homolog of the S100 genes, during zebrafish embryogenesis.
Full-length cDNA coding for the ictacalcin gene, a homolog of the S100 genes, was isolated in zebrafish and mapped on linkage group 16 using the LN54 radiation hybrid panel. The homology and phylogenetic analyses, based on the deduced amino acid sequences, showed the orthologous relationship of ictacalcin genes between zebrafish and other fish species. However, ictacalcin genes constitute an out-group with respect to other members of the S100 gene family. This result supports the findings that fish ictacalcin genes are new members of the S100 gene family and may have evolved after the divergence of teleosts and tetrapods. The zebrafish ictacalcin gene was zygotically transcribed from 12 hours postfertilization onward and was stably expressed throughout adulthood. During zebrafish embryogenesis, the ictacalcin gene was specifically expressed in striated epidermal cells covering the entire embryo. The ictacalcin staining in keratinocytes of striated epithelia was absent in the cytoplasm surrounding the nuclei, but it was highly concentrated in the peripheral margin. Tissues enriched with epithelia folds, such as olfactory epithelium, hatching gland, pectoral fin buds, urogenital opening, and pharynx, showed a robust ictacalcin expression. The strikingly heavy staining of ictacalcin in the pharyngeal region provides us with an early marker to follow the pharynx formation in zebrafish embryos.


[Study of the structure of immunoglobins by small-angle x-ray diffraction. I. The structure of IgMCep in solution].
The data of small-angle X-ray scattering from monoclonal immunoglobulin MCep (IgM) enabled the shape and geometrical parameters of the molecule in solution at 23 degrees C to be established. The molecule is a flat, strongly anisometric particle with radius of gyration 115 A, volume 1,8 X 10(6) A3, maximum size 380 A, thickness 35-40 A. The most probable molecular model in the approximation of homogeneous electron density in the molecule was suggested, its geometry fitting the experimental parameters. The five IgM subunits are located in the equatorial plane, low-electronic-density regions are located in the centre and at the periphery of the macromolecule. In addition, the absence of fixed angle values between Fab-regions in each subunit is indicative of rather high structural mobility at the periphery of the IgM molecule.


Exploring Factors Regarding Transit-related Walking and Walking Duration.
Transit-related walking provides a potential opportunity to promote general walking behavior, yet few studies have examined this issue. Since people's decisions tend to vary as they walk between home and transit and between transit and destination, this study separated trips made in each direction. This study identified the associations between sociodemographics and the 2-step process of transit-related walking: 1) whether transit users walked for home-transit trip or transitdestination trip, and 2) the walking duration for home-transit trip or transit-destination trip among those who walked. This cross-sectional study used the 2009 National Household Travel Survey and used the Heckman 2-step selection model by including 4042 respondents (10,105 trips) who walked all portions for home-transit trip and 3756 (8075 trips) for transitdestination trip. The mean walking duration for home-transit trips (7.60 minutes) was shorter than transit-destination trips (7.87 minutes). Hispanics were more likely to walk for both directions and had higher walking durations than did whites. Respondents living in low-income households were more likely to walk for home-transit trip, but not for transit-destination trips. This study illustrated several implications regarding to transit-related walking, such as creating short home-transit distances and targeting whites in promoting transit-related walking.


Immunolocalization of a human basal epithelium specific keratin in benign and malignant breast disease.
This report describes the immunocytochemical localization of a human basal- or myoepithelial-specific anti-keratin antibody in benign and malignant breast disease. Reactivity patterns with this antibody have demonstrated the lack of myoepithelial or basal epithelial participation in most benign breast specimens examined including those displaying cystic disease, fibrosis, or hyperplasia. However, in specimens of sclerosing adenosis, strong reactivity with the majority of cells in most ducts suggests a major participation of the myoepithelial cell type. Analysis of 118 breast carcinoma specimens has demonstrated strong, homogeneous reactivity in 4% of the specimens, suggesting a role for the basal epithelial cell in malignancy of the human mammary gland and implications for the prognosis of such tumors. Antigenic characterization of the malignant and benign mammary specimens which are uniformly reactive with the antibody has demonstrated the presence of a 51 kd keratin polypeptide not found in the non-reactive specimens.


Physical Determinants of Division 1 Collegiate Basketball, Women's National Basketball League, and Women's National Basketball Association Athletes: With Reference to Lower-Body Sidedness.
Spiteri, T, Binetti, M, Scanlan, AT, Dalbo, VJ, Dolci, F, and Specos, C. Physical determinants of Division 1 Collegiate basketball, Women's National Basketball League, and Women's National Basketball Association athletes: with reference to lower-body sidedness. J Strength Cond Res 33(1): 159-166, 2019-In female basketball, the assumed components of success include power, agility, and the proficiency at executing movements using each limb. However, the importance of these attributes in discriminating between playing levels in female basketball has yet to be determined. The purpose of this study was to compare lower-body power, change of direction (COD) speed, agility, and lower-body sidedness between basketball athletes participating in Division 1 Collegiate basketball (United States), Women's National Basketball League (WNBL) (Australia), and Women's National Basketball Association (WNBA) (United States). Fifteen female athletes from each league (N = 45) completed a double- and single-leg countermovement jump (CMJ), static jump, drop jump, 5-0-5 COD test, and an offensive and a defensive agility test. One-way analysis of variance with post hoc comparisons were conducted to compare differences in physical characteristics (height, body mass, age) and performance outcomes (jump, COD, agility assessments) between playing levels. Separate dependent t-tests were performed to compare lower-body sidedness (left vs. right lower limbs) during the single-leg CMJ jumps (vertical jump height) and 5-0-5 COD test for each limb within each playing level. WNBA athletes displayed significantly greater lower-body power (p = 0.01-0.03) compared with WNBL athletes, significantly faster COD speed (p = 0.02-0.03), and offensive and defensive agility performances (p = 0.02-0.03) compared with WNBL and Collegiate athletes. The WNBL athletes also produced a faster defensive agility performance compared with Collegiate athletes (p = 0.02). Furthermore, WNBA and WNBL athletes exhibited reduced lower-body sidedness compared with Collegiate athletes. These findings indicate the importance of lower-body power, agility, and reduced lower-body imbalances to execute more proficient on-court movements required to compete at higher playing levels.


Specific interaction of polypyrimidine tract-binding protein with the extreme 3'-terminal structure of the hepatitis C virus genome, the 3'X.
We previously identified a highly conserved 98-nucleotide (nt) sequence, the 3'X, as the extreme 3'-terminal structure of the hepatitis C virus (HCV) genome (T. Tanaka, N. Kato, M.-J. Cho, and K. Shimotohno, Biochem. Biophys. Res. Commun. 215:744-749, 1995). Since the 3' end of positive-strand viral RNA is the initiation site of RNA replication, the 3'X should contribute to HCV negative-strand RNA synthesis. Cellular factors may also be involved in this replication mechanism, since several cellular proteins have been shown to interact with the 3'-end regions of other viral genomes. In this study, we found that both 38- and 57-kDa proteins in the human hepatocyte line PH5CH bound specifically to the 3'-end structure of HCV positive-strand RNA by a UV-induced cross-linking assay. The 57-kDa protein (p57), which had higher affinities to RNA probes, recognized a 26-nt sequence including the 5'-terminal 19 nt of the 3'X and 7 flanking nt, designated the transitional region. This sequence contains pyrimidine-rich motifs and shows similarity to the consensus binding sequence of the polypyrimidine tract-binding protein (PTB), which has been implicated in alternative pre-mRNA splicing and cap-independent translation. We found that this 3'X-binding p57 is identical to PTB. The 3'X-binding p57 was immunoprecipitated by anti-PTB antibody, and recombinant PTB bound to the 3'X RNA. In addition, p57 bound solely to the 3'-end region of positive-strand RNA, not to this region of negative-strand RNA. We suggest that 3'X-PTB interaction is involved in the specific initiation of HCV genome replication.


A physiologically based model of 1-nitropyrene metabolism after inhalation or ingestion.
The nitrogen-containing polycyclic aromatic hydrocarbon 1-nitropyrene (NP), a bacterial mutagen and mammalian carcinogen, is a ubiquitous environmental pollutant. A physiologically based toxicokinetic model was developed describing the disposition of NP after oral administration (ingestion) or after inhalation. The model incorporated the following compartments: blood, upper respiratory tract, lung, liver, kidney, gastrointestinal (GI) tract, and a general tissue compartment. First-order rate constants for absorption of NP from the GI tract (2 h-1), metabolism by the liver (30 h-1), excretion of metabolites in bile and urine (2 and 4 h-1, respectively), and covalent binding of NP metabolites to tissue macromolecules (0.05, 0.05, 0.1, and 0.001 h-1 for lung, liver, kidney, and general tissue compartment, respectively) determined from model simulations were used to describe absorption, biotransformation, and excretion of NP. Physiological parameters such as alveolar ventilation, cardiac output, blood flow to organs volume, and tissue/blood partition coefficients described movement of NP and metabolites among compartments. Model predictions for concentrations of NP and metabolites in tissues were compared to experimentally determined data obtained in rats after inhalation of NP. Model predictions for concentrations of NP metabolites covalently bound to tissue macromolecules agreed with experimentally determined data in rats. Levels of bound material in lung and liver were about one-tenth that found in kidney. Results indicated that NP movement among tissue compartments could be described to a large extent by blood flow and organ volume alone (e.g., tissue/blood partition coefficients = 1). The use of physiologically realistic parameters will enable scaling of the model developed using animal studies to predict disposition of NP in humans.


Genome sequence of the cyanobacterium Prochlorococcus marinus SS120, a nearly minimal oxyphototrophic genome.
Prochlorococcus marinus, the dominant photosynthetic organism in the ocean, is found in two main ecological forms: high-light-adapted genotypes in the upper part of the water column and low-light-adapted genotypes at the bottom of the illuminated layer. P. marinus SS120, the complete genome sequence reported here, is an extremely low-light-adapted form. The genome of P. marinus SS120 is composed of a single circular chromosome of 1,751,080 bp with an average G+C content of 36.4%. It contains 1,884 predicted protein-coding genes with an average size of 825 bp, a single rRNA operon, and 40 tRNA genes. Together with the 1.66-Mbp genome of P. marinus MED4, the genome of P. marinus SS120 is one of the two smallest genomes of a photosynthetic organism known to date. It lacks many genes that are involved in photosynthesis, DNA repair, solute uptake, intermediary metabolism, motility, phototaxis, and other functions that are conserved among other cyanobacteria. Systems of signal transduction and environmental stress response show a particularly drastic reduction in the number of components, even taking into account the small size of the SS120 genome. In contrast, housekeeping genes, which encode enzymes of amino acid, nucleotide, cofactor, and cell wall biosynthesis, are all present. Because of its remarkable compactness, the genome of P. marinus SS120 might approximate the minimal gene complement of a photosynthetic organism.


Isoform-selective disruption of AKAP-localized PKA using hydrocarbon stapled peptides.
A-kinase anchoring proteins (AKAPs) play an important role in the spatial and temporal regulation of protein kinase A (PKA) by scaffolding critical intracellular signaling complexes. Here we report the design of conformationally constrained peptides that disrupt interactions between PKA and AKAPs in an isoform-selective manner. Peptides derived from the A Kinase Binding (AKB) domain of several AKAPs were chemically modified to contain an all-hydrocarbon staple and target the docking/dimerization domain of PKA-R, thereby occluding AKAP interactions. The peptides are cell-permeable against diverse human cell lines, are highly isoform-selective for PKA-RII, and can effectively inhibit interactions between AKAPs and PKA-RII in intact cells. These peptides can be applied as useful reagents in cell-based studies to selectively disrupt AKAP-localized PKA-RII activity and block AKAP signaling complexes. In summary, the novel hydrocarbon-stapled peptides developed in this study represent a new class of AKAP disruptors to study compartmentalized RII-regulated PKA signaling in cells.


Intralesional steroid injection therapy in the management of resistant gastrointestinal strictures.
Esophageal strictures are a problem frequently encountered by gastroenterologists. Dilation has been the customary treatment for benign esophageal strictures, and dilation techniques have advanced over the years. Depending on their characteristics and the response to treatment, esophageal strictures can be classified into two types: 1, simple (Schatzki rings, webs, peptic injury, and following sclerotherapy) - these are easily amenable to dilation, with a low recurrence rate after initial treatment; and 2, complex (caused by caustic ingestion, radiation injury, anastomotic strictures, and photodynamic therapy) - these are difficult to dilate and are associated with higher recurrence rates. Refractory strictures are those in which it is not possible to relieve the anatomic restriction successfully up to a diameter of 14 mm over five sessions at 2-weekly intervals, due to cicatricial luminal compromise or fibrosis; and recurrent strictures are those in which it is not possible to maintain a satisfactory luminal diameter for 4 wk once the target diameter of 14 mm has been achieved. There are no standard recommendations for the management of refractory strictures. The various techniques used include intralesional steroid injection combined with dilation; endoscopic incisional therapy, with or without dilation; placement of self-expanding metal stents, Polyflex stents, or biodegradable stents; self-bougienage; and endoscopic surgery. This review discusses the indications, technique, results, and complications of the use of intralesional steroid injections combined with dilation and endoscopic incisional therapy with dilation in refractory strictures.


Expression of microRNAs in human post-mortem amyotrophic lateral sclerosis spinal cords provides insight into disease mechanisms.
Amyotrophic lateral sclerosis is a late-onset and terminal neurodegenerative disease. The majority of cases are sporadic with unknown causes and only a small number of cases are genetically linked. Recent evidence suggests that post-transcriptional regulation and epigenetic mechanisms, such as microRNAs, underlie the onset and progression of neurodegenerative disorders; therefore, altered microRNA expression may result in the dysregulation of key genes and biological pathways that contribute to the development of sporadic amyotrophic lateral sclerosis. Using systems biology analyses on postmortem human spinal cord tissue, we identified dysregulated mature microRNAs and their potential targets previously implicated in functional process and pathways associated with the pathogenesis of ALS. Furthermore, we report a global reduction of mature microRNAs, alterations in microRNA processing, and support for a role of the nucleotide binding protein, TAR DNA binding protein 43, in regulating sporadic amyotrophic lateral sclerosis-associated microRNAs, thereby offering a potential underlying mechanism for sporadic amyotrophic lateral sclerosis.


Dissecting the plant transcriptome and the regulatory responses to phosphate deprivation.
Inorganic phosphate (Pi) is an essential nutrient for plants, and the low bioavailability of Pi in soils is often a limitation to growth and development. Consequently, plants have evolved a range of regulatory mechanisms to adapt to phosphorus-starvation in order to optimise uptake and assimilation of Pi. Recently, significant progress has been made in elucidating these mechanisms. The coordinated expression of a large number of genes is important for many of these adaptations. Several global expression studies using microarray analysis have been conducted in Arabidopsis thaliana. These studies provide a valuable basis for the identification of new regulatory genes and promoter elements to further the understanding of Pi-dependent gene regulation. With focus on the Arabidopsis transcriptome, we extract common findings that point to new groups of putative regulators, including the NAC, MYB, ethylene response factor/APETALA2, zinc-finger, WRKY and CCAAT-binding families. With a number of new discoveries of regulatory elements, a complex regulatory network is emerging. Some regulatory elements, e.g. the transcription factor PHR1 and the microRNA (miRNA) miR399 and associated factors are well documented, yet not fully understood, whereas other suggested components need further characterisation. Here, we evaluate the contribution of the regulatory elements to the P-responses and present a model comprising factors directly or indirectly involved in transcriptional regulation and the role of miRNAs as regulators and long-distance signals. A striking feature is a series of feedback loops and parallel mechanisms that can modify and attenuate responses. We suggest that these mechanisms are instrumental in providing an accurate response and in keeping P-homeostasis.


Rationalized DNA sequencing-based protocol for genotyping patients receiving coumarin therapy.
During the last decade genetic factors affecting coumarin therapy have been extensively investigated. The most important genes appear to be CYP2C9 and VKORC1, and different studies have shown that DNA testing can dramatically improve the safety and effectiveness of the therapy. However, the implementation of pharmacogenetic testing in everyday practice is still not a reality. Facilities and ability to get results before the start of therapy are very important. The implementation of specific methodology and equipment for particular type of diagnostics can represent a serious, even impossible, financial hurdle to overcome (especially in developing countries). For this reason, the use of every tool that contributes to rationalization of the existing methods can be a considerable asset. Therefore, we set the goal to rationalize our current DNA sequencing based protocol for analysis of the VKORC1 c.-1639G> A, CYP2C9*2 and CYP2C9*3 variant alleles, in order to obtain shorter and easier procedure. Simplification of the protocol was achieved by setting up multiplex PCR and omitting DNA extraction. This rationalization of the existing DNA sequencing based procedure allows getting results in 12 hours. The new protocol was tested on 118 samples. Obtained results have shown full accordance to those obtained with previous, non-modified protocol. Therefore, given the circumstances, we consider that protocol for pharmocogenetic testing should be made more accessible - both to doctors and patients. It is one of the prerequisites in order to make genotyping prior to the therapy common practice.


Inability of the activated partial thromboplastin time to predict heparin levels. Time to reassess guidelines for heparin assays.
In treating venous thromboembolic disorders, patient outcomes appear to correlate with heparin levels. Due to pharmacokinetic and pharmacodynamic variations, a relationship between heparin dose and level cannot be reliably predicted in individual patients. Some patients have low heparin levels despite therapeutic activated partial thromboplastin times (aPTTs), which may increase their risk for recurrent thromboembolism. Patients with high heparin requirements appear to have fewer bleeding episodes with heparin level-guided therapy. The aPTT does not reliably correlate with heparin blood concentrations or antithrombotic effects. Consequently, heparin therapy monitored with heparin levels may be more effective and safer. To prospectively determine whether (1) the aPTT therapeutic range adequately predicts heparin levels in 38 patients used to establish the therapeutic aPTT range as is currently recommended and (2) whether 3 paired sets of aPTT-antifactor Xa levels provide the basis for using aPTTs to predict subsequent heparin levels in individual patients (n = 27) receiving intravenous heparin for coronary artery disease or venous thromboembolic disease. In the therapeutic aPTT range established, the R2 value for the relationship was 0.4. Prediction intervals were wide. For an aPTT of 60 seconds, the 95% prediction interval estimates were heparin levels of 0.05 to 1.0 U/mL. In individual patients, the aPTT-antifactor Xa relationship had an average R2 value of 0.75. There was no consistent relationship between the aPTT and anti-factor Xa level in a significant number of patients. The aPTT does not appear to be a useful surrogate for heparin levels. These findings suggest that the current recommendations on the use of heparin levels should be expanded.


Alterations in Metabolic Status and Headshaking Behavior Following Intravenous Administration of Hypertonic Solutions in Horses with Trigeminal-Mediated Headshaking.
Trigeminal-nerve-mediated headshaking represents a major welfare challenge for owners and veterinarians and is caused by a low threshold firing of the trigeminal nerve resulting in pain manifested as violent head jerking that often terminates the horse's career and life due to poor quality of life and suffering. As metabolic changes such as acid⁻base status and electrolytes play a role in nerve firing, this study sought to assess the effects following administration of hypertonic solutions on headshaking behavior in affected horses. This prospective randomized controlled crossover design utilized six horses affected with trigeminal-mediated headshaking and three treatment groups receiving intravenous administration of 5% dextrose solution at 2 mL/kg bwt (DS), NaCl 7.5% at 4 mL/kg bwt (HS), or NaHCO₃ 8.4% at 2 mmol/kg bwt (HB). Horses were assessed for headshaking behavior changes at times T0 (baseline, before infusion) and T15, 30, 60, 120 min post infusion. Venous blood variables: pH, HCO₃−, standard base excess (SBE), Na⁺, Cl−, K⁺, Ca2+, Mg2+, total magnesium (tMg), glucose, and lactate were measured at T0 (baseline, before infusion) and T5, 15, 30, 60, 120 min post infusion. Strong ion difference (SID) and anion gap (AG) were calculated for each time point. With HB treatment, there was greater than 50% reduction in headshaking rate. There was an effect of time on increasing headshaking rate. There was an effect of breed on headshaking rate. Changes in blood parameters following DS were virtually absent. Infusion of HS caused mild changes and did not vary much from baseline except for SID and AG. Only infusion of HB caused blood pH and HCO₃− to be outside of the physiologic range (alkalemia and metabolic alkalosis, respectively), SBE to double or triple, AG to decrease, and SID to increase compared to baseline. Infusion of DS was followed by increase in blood glucose and decrease in blood Na⁺. Infusion of HS was followed by increase in Na⁺ and Cl− and decrease in Mg2+. Infusion of HB was followed by decrease in Mg2+. Blood tMg, K⁺, and Ca2+ decreased slightly, but did not vary greatly from baseline following any of the treatments, remaining within physiologic ranges. Changes in blood composition were transient. Among all treatments, only HB had an effect on headshaking rate. The limited effects following these fluids were likely due to normal mechanisms of regulation of blood levels of pH and electrolytes. Further investigations of changes in electrolytes that might affect nerve firing should be explored.


Comparisons of predicted genetic modules: identification of co-expressed genes through module gene flow.
A question of fundamental importance is the definition and identification of modules from microarray experiments. A wide variety of techniques have been used to gain insight into the elucidation of such modules. One problem, however, is the inability to directly compare results between the different data sets produced due to the inherent parameterizations of their approaches. We first aim to provide a mechanism by which different approaches to module finding can be directly compared. Moreover, the same approach can be used to internally compare the modules predicted by the same technique, but at different parameterizations. We apply this approach to analyze the flow of genes through modules at different module thresholds of the Barkai Signature method, thereby further resolving the modules into sets of co-expressed genes.


[ICU-acquired neuromyopathy, delirium and sedation in intensive care unit].
ICU-acquired neuromyopathy (NMAR) and delirium are the two most frequent and severe neurological complications of intensive care medicine. Their mechanisms still remain to be elucidated. The objective of this review is to address the potential role of sedation in occurrence of these complications. There is no evidence that sedation is involved in NMARs. However, the hypothesis that muscle inactivity induced by sedation fosters NMAR is an argument to discontinue or reduce sedatives infusion whenever possible. It is also recommended not to administer propofol more than 48 h at an infusion rate above 5 mg/kg per hour in patients with systemic inflammatory response syndrome, because of the risk of propofol infusion syndrome, which includes notably rhabdomyolysis. The relationship between delirium and sedation are controversial because in most studies, patients were considered delirious though being still sedated and multivariate analysis was lacking. One study showed that lorazepam given continuously was an independent risk factor for daily transition to delirium 24 h later with a 20% increase risk of every unit dose (expressed as log(e)mg). The impact of deepness, daily interruption or titration of sedation on the prevalence of delirium has never been assessed but it seems that deep sedation has to be avoided.


Detection of extracellular RNAs in cancer and viral infection via tethered cationic lipoplex nanoparticles containing molecular beacons.
Noninvasive early detection methods have the potential to reduce mortality rates of both cancer and infectious diseases. Here, we present a novel assay by which tethered cationic lipoplex nanoparticles containing molecular beacons (MBs) can capture cancer cell-derived exosomes or viruses and identify encapsulated RNAs in a single step. A series of ultracentrifugation and Exoquick isolation kit were first used to isolate exosomes from the cell culture medium and human serum, respectively. Cationic lipoplex nanoparticles linked onto the surface of a thin glass plate capture negatively charged viruses or cell-secreted exosomes by electrostatic interactions to form larger nanoscale complexes. Lipoplex/virus or lipoplex/exosome fusion leads to the mixing of viral/exosomal RNAs and MBs within the lipoplexes. After the target RNAs specially bind to the MBs, exosomes enriched in target RNAs are readily identified by the fluorescence signals of MBs. The in situ detection of target extracellular RNAs without diluting the samples leads to high detection sensitivity not achievable by existing methods, e.g., quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Here we demonstrate this concept using lentivirus and serum from lung cancer patients.


Eosinophilic cystitis associated with Glanzmann's thrombasthenia. A case report.
Eosinophilic cystitis is a rare condition, only 41 cases having been recorded in the literature. Glanzmann's thrombasthenia has been documented more than 100 times. The presence of these two conditions in one patient has, to our knowledge, not yet been reported in the English-language literature.


Immunomodulators targeting MARCO expression improve resistance to postinfluenza bacterial pneumonia.
Downregulation of the alveolar macrophage (AM) receptor with collagenous structure (MARCO) leads to susceptibility to postinfluenza bacterial pneumonia, a major cause of morbidity and mortality. We sought to determine whether immunomodulation of MARCO could improve host defense and resistance to secondary bacterial pneumonia. RNAseq analysis identified a striking increase in MARCO expression between days 9 and 11 after influenza infection and indicated important roles for Akt and Nrf2 in MARCO recovery. In vitro, primary human AM-like monocyte-derived macrophages (AM-MDMs) and THP-1 macrophages were treated with IFNγ to model influenza effects. Activators of Nrf2 (sulforaphane) or Akt (SC79) caused increased MARCO expression and a MARCO-dependent improvement in phagocytosis in IFNγ-treated cells and improved survival in mice with postinfluenza pneumococcal pneumonia. Transcription factor analysis also indicated a role for transcription factor E-box (TFEB) in MARCO recovery. Overexpression of TFEB in THP-1 cells led to marked increases in MARCO. The ability of Akt activation to increase MARCO expression in IFNγ-treated AM-MDMs was abrogated in TFEB-knockdown cells, indicating Akt increases MARCO expression through TFEB. Increasing MARCO expression by targeting Nrf2 signaling or the Akt-TFEB-MARCO pathway are promising strategies to improve bacterial clearance and survival in postinfluenza bacterial pneumonia.


CTL effector function within the central nervous system requires CD4+ T cells.
CTL responses induced during most viral infections are independent of help derived from the CD4+ T cell population. However, clearance of virus from the central nervous system (CNS) during infection with the neurotropic JHM strain of mouse hepatitis virus is inhibited in the absence of CD4+ T cells. Adoptive transfer of activated CD8+ T cells with virus-specific cytolytic activity into CD4+ T cell-depleted hosts demonstrated that CD4+ T cells were one component of the host response required for expression of CTL effector function(s) within the CNS. Analysis of mice infected with the JHM strain of mouse hepatitis virus demonstrated that, in contrast to CD8+ T cells, few CD4+ T cells entered the brain parenchyma. Although fewer CD8+ T cells entered the brain parenchyma in mice depleted of CD4+ T cells, access of CTL was not inhibited in the absence of CD4+ T cells. The number of apoptotic lymphocytes in the CNS increased in the absence of CD4+ T cells, suggesting that CTL enter the CNS during viral infection in a CD4-independent manner. However, these cells rapidly undergo apoptosis, indicating that expression of CTL effector function with the parenchyma of the CNS is CD4 dependent. These data raise the possibility that programmed cell death of CD8+ T cells within the CNS is due to the increased Ag present in the CNS of infected CD4 depleted mice or that autocrine cytokines, which maintain CTL activity within peripheral tissues, are inhibited in the microenvironment of the CNS.


Economic and clinical effects of evaluating rapid viral response to peginterferon alfa-2b plus ribavirin for the initial treatment of chronic hepatitis C.
Evaluation of 12-wk viral response to initial antiviral therapy for chronic hepatitis C has been recommended to minimize antiviral-associated morbidity and costs. The aim of this study was to examine the economic and clinical effects of evaluating rapid viral response during antiviral therapy for treatment naive chronic hepatitis C patients. We applied viral response and drug dosage from an international randomized clinical trial of ribavirin plus peginterferon alfa-2b or ribavirin plus interferon alfa-2b to a previously published computer cohort simulation to project lifelong clinical and economic outcomes. Natural history and economic estimates were based on published literature, expert panel estimates, and actual variable and reimbursement cost data. The assessment of 12-wk rapid viral response reduced antiviral treatment duration by 40-44% and antiviral costs by 44-45% (savings of $15,116-16,268 for peginterferon plus ribavirin and $8300 for interferon plus ribavirin) compared to full 48-wk dosing. With the 12-wk evaluation, the marginal cost-effectiveness of peginterferon plus ribavirin versus interferon plus ribavirin was $13,600-22,800 compared with $14,600-25,000 per discounted quality adjusted life-year gained with the 24-wk evaluation. For genotype 1, hepatitis C infected patients, 12-wk testing for peginterferon plus ribavirin remaining preferred and cost-effective compared with interferon plus ribavirin. For genotype 2 or 3, hepatitis C infected patients, 12-wk testing yielded similar results to those of 24-wk treatment. Assessment of 12-wk viral response in genotype 1, hepatitis C infected patients should reduce peginterferon plus ribavirin morbidity and costs and improve its cost-effectiveness; however, for genotype 2 and 3, hepatitis C infected patients, 12-wk testing and 24-wk treatment have similar outcomes. Decisions regarding continuation of antiviral treatment should also consider the variability in the accuracy of quantitative viral assays as well as patient preferences and other potential benefits of the same treatments.


Learning outcomes of leader and leadership development: Method of Soft and Female leadership (SFL) empirical framework for driving the gender equality change in Mediterranean Engineering Education.
The foundation of gender equality was built some decades ago, but higher education institutions are far from achieving it. Perhaps the leadership needs to integrate new narratives for a greater commitment especially in engineering, and new tools for the existing toolbox. This study aims to share the outcomes of soft female leadership framework (SFL) development for gender equality at the RMEI network, entailing the commitment of top-managers from engineering schools, creation of new leaders to lead the change at their institutions, students acting as change-agents, and an active community of practice. The SFL toolbox comprises self-awareness, humanistic care, intuition, creativity, and trust. The transformation of mindset, skillset, and culture entails using Sustainable Development Goals (SDGs), 'Systemic View of Life' arguments, and drawing knowledge from organizational learning frameworks, scientific phenomena, and mechanisms, such as a) 4I-Intuiting-Interpreting-Integrating-Institutionalizing organizational learning process starting from intuition to achieve an institutional change; b) 'Stigmergy' scientific mechanism of self-organized collective schemes with coordinated actions and interactions, in which the action performed by an agent leaves a trace in the environment that stimulates subsequent actions; c) 'Spillover' phenomenon advocating that the behavior of an agent can bring the adoption of related behaviors by other agents. The RMEI gender equality plan (GEP) was evaluated by the HORIZON2020 TARGET project consortium. The SFL excelled as successful in setting goals, articulating a policy that integrates systems approach frameworks, insights from science and technology, innovation, ecology, philosophy, self-awareness, ethics, and values. The Covid-19 pandemic disrupted physical meetings, but the process of change was not ceased at the network because we disrupted the disruption by boosting collaborative knowledge, consolidation and dissemination processes. The SFL framework integrates context, regional, and temporal characteristics, alongside cognitive, affective, and motivational outcomes over behavioral outcomes, new mindsets beyond organizational skills, and collaborative learning over individual learning.


Antidepressant-like and anxiolytic-like effects of cannabidiol: a chemical compound of Cannabis sativa.
Anxiety and depression are pathologies that affect human beings in many aspects of life, including social life, productivity and health. Cannabidiol (CBD) is a constituent non-psychotomimetic of Cannabis sativa with great psychiatric potential, including uses as an antidepressant-like and anxiolytic-like compound. The aim of this study is to review studies of animal models using CBD as an anxiolytic-like and antidepressant-like compound. Studies involving animal models, performing a variety of experiments on the above-mentioned disorders, such as the forced swimming test (FST), elevated plus maze (EPM) and Vogel conflict test (VCT), suggest that CBD exhibited an anti-anxiety and antidepressant effects in animal models discussed. Experiments with CBD demonstrated non-activation of neuroreceptors CB1 and CB2. Most of the studies demonstrated a good interaction between CBD and the 5-HT1A neuro-receptor.


ALK alterations and inhibition in lung cancer.
The advent of precision medicine in non-small cell lung cancer has remarkably altered the direction of research and improved clinical outcomes. The identification of molecular subsets with differential response to targeted therapies began with the identification of epidermal growth factor receptor mutated tumors in subsets of non-small cell lung cancer (NSCLC). Emboldened by unprecedented response rates to kinase inhibitors seen in that subset, the oncologic community searched for other molecular subsets featuring oncogene addiction. An early result of this search was the discovery of NSCLC driven by activating rearrangements of the anaplastic lymphoma kinase (ALK) gene. In an astoundingly brief period following the recognition of ALK-positive NSCLC, details of the biology, clinicopathologic features, development of targeted inhibitors, mechanisms of therapeutic resistance, and new generations of treatment were elucidated. This review summarizes the current understanding of the pathologic features, diagnostic approach, treatment options, resistance mechanisms, and future research areas for ALK-positive NSCLC.


High-energy phosphate transfer in human muscle: diffusion of phosphocreatine.
The creatine kinase (CK) reaction is central to muscle energetics, buffering ATP levels during periods of intense activity via consumption of phosphocreatine (PCr). PCr is believed to serve as a spatial shuttle of high-energy phosphate between sites of energy production in the mitochondria and sites of energy utilization in the myofibrils via diffusion. Knowledge of the diffusion coefficient of PCr (D(PCr)) is thus critical for modeling and understanding energy transport in the myocyte, but D(PCr) has not been measured in humans. Using localized phosphorus magnetic resonance spectroscopy, we measured D(PCr) in the calf muscle of 11 adults as a function of direction and diffusion time. The results show that the diffusion of PCr is anisotropic, with significantly higher diffusion along the muscle fibers, and that the diffusion of PCr is restricted to a ∼28-μm pathlength assuming a cylindrical model, with an unbounded diffusion coefficient of ∼0.69 × 10(-3) mm(2)/s. This distance is comparable in size to the myofiber radius. On the basis of prior measures of CK reaction kinetics in human muscle, the expected diffusion distance of PCr during its half-life in the CK reaction is ∼66 μm. This distance is much greater than the average distances between mitochondria and myofibrils. Thus these first measurements of PCr diffusion in human muscle in vivo support the view that PCr diffusion is not a factor limiting high-energy phosphate transport between the mitochondria and the myofibrils in healthy resting myocytes.


Psychiatric education in the correctional setting: challenges and opportunities.
As the need for mental healthcare services within correctional settings in the US increases, so does the need for a mental health workforce that is motivated to work within such systems. One potentially effective method by which to increase the number of psychiatrists working in jails, prisons, and parole clinics is to provide exposure to these environments during their training. Correctional settings can serve as unique training sites for medical students and psychiatric residents and fellows. Such training experiences can provide a host of benefits to both trainees and staff within the correctional mental health system. Alongside many potential benefits exist substantial potential barriers to coordinating correctional training experiences, including both programme directors' and residents' concerns regarding safety and enjoyment and negative perceptions of inmate and prisoner patients. The establishment of academic affiliations with correctional institutions and didactic instruction on commonly encountered clinical issues with inmate populations may be methods of diffusing these concerns. Improving residents' and fellows' training experiences offers a hope for increasing the attractiveness of a career in correctional psychiatry.


Suppression of electron leakage by inserting a thin undoped InGaN layer prior to electron blocking layer in InGaN-based blue-violet laser diodes.
InGaN-based blue-violet laser diodes (LDs) suffer from electron leakage into the p-type regions, which could be only partially alleviated by employing the electron blocking layer (EBL). Here, a thin undoped InGaN interlayer prior to EBL is proposed to create an additional forbidden energy range above the natural conduction band edge, which further suppresses the electron leakage and thus improve the characteristics of LDs. Numerical device simulations reveal that when the proper composition and thickness of InGaN interlayer are chosen, the electron leakage could be efficiently eliminated without inducing any severe accumulation of electrons at the interlayer, resulting in a maximum output power of the device.


Insulin resistance is associated with an exaggerated blood pressure response to ischemic rhythmic handgrip exercise in nondiabetic older adults.
Patients with type 2 diabetes display an exaggerated pressor response to exercise. However, evidence supporting the association between the magnitude of the pressor response to exercise and insulin resistance-related factors including hemoglobin A1c (HbA1c) or homeostatic model assessment of insulin resistance (HOMA-IR) in nondiabetic subjects has remained sparse and inconclusive. Thus we investigated the relationship between cardiovascular responses to exercise and insulin resistance-related factors in nondiabetic healthy men (n = 23) and women (n = 22) above 60 yr old. We measured heart rate (HR) and blood pressure (BP) responses during: isometric handgrip (IHG) exercise of 30% maximal voluntary contraction, a period of skeletal muscle ischemia (SMI) induced by tourniqueting the arm after IHG, and rhythmic dynamic handgrip (DHG) exercise during SMI. Greater diastolic BP (DBP) responses to DHG with SMI was associated with male sex (r = 0.44, P = 0.02) and higher HbA1c (r = 0.33, P = 0.03), heart-ankle pulse wave velocity (haPWV) (r = 0.45, P < 0.01), and resting systolic BP (SBP) (r = 0.36, P = 0.02). HbA1c persisted as a significant determinant explaining the variance in the DBP response to DHG with SMI in multivariate models despite adjustment for sex, haPWV, and resting SBP. It was also determined that the DBP response to DHG with SMI in a group in which HOMA-IR was abnormal (Δ33 ± 3 mmHg) was significantly higher than that of groups in which HOMA-IR was at intermediate (Δ20 ± 4 mmHg) and normal (Δ23 ± 2 mmHg) levels. These data suggest that even in nondiabetic older adults, insulin resistance is related to an exaggerated pressor response to exercise especially when performed under ischemic conditions.NEW & NOTEWORTHY The diastolic blood pressure response to rhythmic dynamic handgrip exercise under ischemic conditions was demonstrated to be correlated with insulin resistance-related factors in nondiabetic older adults. This finding provides important insight to the prescription of exercise in this particular patient population as the blood pressure response to exercise, especially under ischemic conditions, could be exaggerated to nonsafe levels.


Naringenin confers protection against experimental autoimmune encephalomyelitis through modulating the gut-brain axis: A multiomics analysis.
Multiple sclerosis (MS) is a disease of the central nervous system that involves the immune system attacking the protective covering of nerve fibers. This disease can be influenced by both environmental and genetic factors. Evidence has highlighted the critical role of the intestinal microbiota in MS and its animal model, experimental autoimmune encephalomyelitis (EAE). The composition of gut microflora is mainly determined by dietary components, which, in turn, modulate host homeostasis. A diet rich in naringenin at 0.5% can effectively mitigate the severity of EAE in mice. However, there is little direct data on the impact of naringenin at optimal doses on EAE development, as well as its intestinal microbiota and metabolites. Our study revealed that 2.0% naringenin resulted in the lowest clinical score and pathological changes in EAE mice, and altered the gene expression profiles associated with inflammation and immunity in spinal cord tissue. We then used untargeted metabolomics and 16S rRNA gene sequences to identify metabolites and intestinal microbiota, respectively. Naringenin supplementation enriched gut microbiota in EAE mice, including increasing the abundance of Paraprevotellaceae and Comamonadaceae, while decreasing the abundance of Deltaproteobacteria, RF39, and Desulfovibrionaceae. Furthermore, the changes in gut microbiota affected the production of metabolites in the feces and brain, suggesting a role in regulating the gut-brain axis. Finally, we conducted a fecal transplantation experiment to validate that gut microbiota partly mediates the effect of naringenin on EAE alleviation. In conclusion, naringenin has potential immunomodulatory effects that are influenced to some extent by the gut microbiome.