A rapid and simple DNA fingerprinting method using RFLP and SSCP analysis of the hypervariable noncoding region of human mitochondrial DNA.
A simple method for identification of individuals and maternity testing has been developed. This technique includes PCR amplification of the 199-bp hypervariable portion of the noncoding region of human mtDNA, digestion with RsaI and subsequent SSCP analysis of restriction fragments with DNA silver staining. Using this approach we have analysed the DNA fingerprint patterns of the family members. The fingerprints of maternally related individuals appeared to be identical in three generations, while maternally unrelated members of the family showed differences in their fingerprints, either in SSCP or both RFLP and SSCP patterns. Sequencing data have confirmed the results obtained. Further DNA fingerprinting analysis of 19 unrelated mother-child pairs by means of the method described revealed complete identity of the fingerprint patterns within the pairs. The probability of a random fingerprint match for two maternally unrelated individuals was estimated as 8%.


Cannabidiol (CBD) Dosing: Plasma Pharmacokinetics and Effects on Accumulation in Skeletal Muscle, Liver and Adipose Tissue.
Oral cannabidiol (CBD) consumption is widespread in North America and Europe, as it has analgesic, neuroprotective and antitumor effects. Although oral CBD consumption in humans affords beneficial effects in epileptic and inflammatory states, its pharmacokinetics and subsequent uptake into tissue are largely unknown. This study investigated plasma pharmacokinetics and accumulation of CBD in gastrocnemius muscle, liver and adipose tissue in adult rats following oral gavage. CBD was fed relative to body mass at 0 (control), 30, 115, or 230 mg/Kg/day for 28 days; with 6 males and 6 females per dosing group. Pharmacokinetics were assessed on day 1 and day 28 in the group receiving CBD at 115 mg/Kg/day. The rise in tissue CBD was closely related to specific pharmacokinetic parameters, and adipose tissue levels were ~10 to ~100 fold greater than liver or muscle. Tissue CBD levels were moderately correlated between adipose and muscle, and adipose and liver, but were highly correlated for liver and muscle. CBD feeding resulted in several gender-specific effects, including changes in pharmacokinetics, relationships between pharmacokinetic parameters and tissue CBD and differences in tissue CBD levels. CBD accumulation in mammalian tissues has the potential to influence receptor binding and metabolism; therefore, the present findings may have relevance for developing oral dosing regimens.


Hybridoma antibodies to the murine coronavirus JHM: characterization of epitopes on the peplomer protein (E2).
A panel of hybridoma antibodies that react with the surface peplomer glycoprotein (E2) of the murine coronavirus JHM were produced to characterize major antigenic domains associated with functions related to virulence. Three groups of hybridoma antibodies were differentiated by immunoprecipitation of lysates from JHM-infected cells. One group precipitated the virion structural proteins gp170 and gp98 together with the intracellular form of E2, gp150. A second group reacted with gp98 and gp150, and a third group precipitated gp150 only. Competition assays with biotinylated hybridoma antibodies allowed the definition of at least six different epitope groups. Only those antibodies which immunoprecipitated both gp170 and gp98 neutralized infectivity, inhibited cell fusion and protected infected rats against acute disease. Another class of antibodies binding to gp170 and gp98 also neutralized JHM virus, but did not inhibit fusion and did not protect against disease. Antibodies that immunoprecipitated gp150 and gp98 revealed only weak neutralization and did not inhibit cell fusion or protect animals. Four epitopes were defined by antibodies that immunoprecipitated gp150, but revealed no biological activity. These data indicate that the site responsible for cell fusion is associated with an epitope group carried by gp170 and gp98. Neutralizing antibodies bind to this and another epitope. Furthermore, protection of JHM-infected rats against acute disease requires both inhibition of cell fusion and neutralization of virus.


A wild goose metapneumovirus containing a large attachment glycoprotein is avirulent but immunoprotective in domestic turkeys.
The genomic structure and composition of an avian metapneumovirus (aMPV) recently isolated from wild Canada geese (goose 15a/01) in the United States, together with its replication, virulence, and immunogenicity in domestic turkeys, were investigated. The sizes of seven of the eight genes, sequence identity, and genome organization of goose aMPV were similar to those of turkey aMPV subtype C (aMPV/C) strains, indicating that it belonged to the subtype. However, the goose virus contained the largest attachment (G) gene of any pneumovirus or metapneumovirus, with the predicted G protein of 585 amino acids (aa) more than twice the sizes of G proteins from other subtype C viruses and human metapneumovirus and more than 170 aa larger than the G proteins from the other aMPV subtypes (subtypes A, B, and D). The large G gene resulted from a 1,015-nucleotide insertion at 18 nucleotides upstream of the termination signal of the turkey aMPV/C G gene. Three other aMPV isolates from Canada geese had similarly large G genes, whereas analysis of recent aMPV strains circulating in U.S. turkeys did not indicate the presence of the goose virus-like strain. In vitro, the goose virus replicated to levels (2 x 10(5) to 5 x 10(5) 50% tissue culture infective dose) comparable to those produced by turkey aMPV/C strains. More importantly, the virus replicated efficiently in the upper respiratory tract of domestic turkeys but with no clinical signs in either day-old or 2-week-old turkeys. The virus was also horizontally transmitted to naïve birds, and turkey infections with goose 15a/01 induced production of aMPV-specific antibodies. Challenging day-old or 2-week-old turkeys vaccinated with live goose aMPV resulted in lower clinical scores in 33% of the birds, whereas the rest of the birds had no detectable clinical signs of the upper respiratory disease, suggesting that the mutant virus may be a safe and effective vaccine against aMPV infection outbreaks in commercial turkeys.


High-Throughput Assay and Discovery of Small Molecules that Interrupt Malaria Transmission.
Preventing transmission is an important element of malaria control. However, most of the current available methods to assay for malaria transmission blocking are relatively low throughput and cannot be applied to large chemical libraries. We have developed a high-throughput and cost-effective assay, the Saponin-lysis Sexual Stage Assay (SaLSSA), for identifying small molecules with transmission-blocking capacity. SaLSSA analysis of 13,983 unique compounds uncovered that >90% of well-characterized antimalarials, including endoperoxides and 4-aminoquinolines, as well as compounds active against asexual blood stages, lost most of their killing activity when parasites developed into metabolically quiescent stage V gametocytes. On the other hand, we identified compounds with consistent low nanomolar transmission-blocking activity, some of which showed cross-reactivity against asexual blood and liver stages. The data clearly emphasize substantial physiological differences between sexual and asexual parasites and provide a tool and starting points for the discovery and development of transmission-blocking drugs.


A new approach in the first-line treatment of bacterial and mycotic vulvovaginitis with topical lipohydroperoxides and glycyrrhetic acid: a comparative study.
PURPOSE OF INVESTIGATIONS: The aim of this randomized controlled trial was to evaluate efficacy and tolerability of a new association of lipohydroperoxides and glycyrrhetic acid on topical treatment of bacterial and mycotic vulvovaginitis. One hundred consecutive patients with bacterial or mycotic vulvovaginitis were randomly assigned to a study group treated with vaginal lipohydroperoxides and a derivative of glycyrrhetic acid for three days (n = 50), and a control group using vaginal antibacterial metronidazole (500 mg) or antimycotic econazole (150 mg) for six days (n = 50). A clinical and microbiological response was achieved in 80.4% and 88.9% in investigational and control group, respectively (p > 0.05). Compared to traditional antimicrobial drugs, the effect appears to be faster and safer, even if not significantly. The 6-month recurrence rate was 7.7% and 5.6% in the investigational and control group, respectively. Topical medication based on lipohydroperoxides and glycyrrhetic acid showed a clinical and microbiological efficacy in the first-line treatment of bacterial and mycotic vulvovaginitis, comparable to conventional drugs.


The therapeutic alliance--a proteus in disguise.
The therapeutic alliance is a vital component of not only the psychoanalytic relation but of all therapeutic encounters between psychotherapist and patient. Despite the universal application and realization of the alliance concept in therapeutic endeavors, it is often ignored as an operative concept in the therapeutic theoretical armamentarium or is formulated in alternative terms. It also comes into play implicitly, even when the concept is formally dismissed as irrelevant. This discussion addresses the meaning and variations of expression of the alliance in the clinical setting and focuses particularly on ways in which the alliance is actually formulated in alternate terms that usually address some partial aspect of the alliance without acknowledging its relevance or importance in the therapeutic relationship and interaction between therapist and patient. I conclude that even when its role in therapy is ignored, minimized, or denied, the alliance continues to play a vital role that requires therapeutic attention and processing in its own right. (PsycINFO Database Record (c) 2010 APA, all rights reserved).


Glutaraldehyde fixation of the cAMP-dependent Na+/H+ exchanger in trout red cells.
It has been shown that the addition of a beta-adrenergic catecholamine to a trout red blood cell suspension induces a 60-100-fold increase of sodium permeability resulting from the activation of a cAMP-dependent Na+/H+ antiport. Subsequent addition of propranolol almost instantaneously reduces the intracellular cAMP concentration, and thus the Na permeability, to their basal values (Mahé et al., 1985). If glutaraldehyde (0.06-0.1%) is added when the Na+/H+ exchanger is activated after hormonal stimulation, addition of propranolol no longer inhibits Na permeability: once activated and fixed by glutaraldehyde, the cAMP dependence disappears. Glutaraldehyde alone causes a rapid decrease in the cellular cAMP concentration. In its fixed state the antiporter is fully amiloride sensitive. The switching on of the Na+/H+ exchange by cAMP is rapidly (2 min) followed by acute but progressive desensitization of the exchanger (Garcia-Romeu et al., 1988). The desensitization depends on the concentration of external sodium, being maximal at a normal Na concentration (145 mM) and nonexistent at a low Na concentration (20 mM). If glutaraldehyde is added after activation in nondesensitizing conditions (20 mM Na), transfer to a Na-rich medium induces only a very slight desensitization: thus the fixative can "freeze" the exchanger in the nondesensitizing conformation. NO3- inhibits the activity of the cAMP-dependent Na+/H+ antiporter of the trout red blood cell (Borgese et al., 1986). If glutaraldehyde is added when the cells are activated by cAMP in a chloride-containing medium, the activity of the exchanger is no longer inhibited when Cl- is replaced by NO3-. Conversely, after fixation in NO3- medium replacement of NO3- by Cl- has very little stimulatory effect. This indicates that the anion dependence is not a specific requirement for the exchange process but that the anion environment is critical for the switching on of the Na+/H+ exchanger and for the maintenance of its activated configuration.


Analysis of the Orchidaceae Diversity in the Pululahua Reserve, Ecuador: Opportunities and Constraints as Regards the Biodiversity Conservation of the Cloud Mountain Forest.
The Pululahua Geobotanical Reserve is a protected natural area in the cloud mountain forest of Ecuador, so rich in orchid species despite being a volcanic area still under threat of volcanic activity. A comparative biodiversity study of orchids was carried out in two different sectors, Chaupisacha (CH) and La Reventazón (LR). Data were collected in 1 ha plots in each sector, in which all the orchids found were counted and two individuals of each species were retained. Immature individuals were conserved in a plant nursery until flowering. In CH, there were 922 individuals grouped into 24 genera and 55 species, while LR had 9196 individuals grouped into 26 genera and 46 species; only 14 species were found in both sectors. Different density and diversity indexes were calculated. The density (ind./100 m2) of CH was 0.96, while that of LR was 185.92. Simpson's diversity (1 - λ) attained CH 0.903 ± 0.01 and LR 0.85 ± 0.01. The orchid diversity measured by the Shannon-Wiener diversity index (H') was 1.29 for CH, differing significantly from that of LR (H' 1.02). The medium equity (Jaccard's J') found was 0.61 in CH and 90.78 in LR. Limitations as regards the natural dispersion of orchids seemed to favor endemism. Some species, such as Dracula felix and Restrepia guttulate, are threatened with disappearance from the wild or are vulnerable, as is the case for Epidendrum polyanthogastrium. A lack of information on the phenology and anthropic impacts in the area limit the conservation of species, signifying that new protected figures and seed banks are necessary, especially in CH, owing to its high diversity of orchids.


Letter: Adenocarcinoma of the lung associated with pityriasis rubra pilaris.
We describe a case of pityriasis rubra pilaris refractory to conventional treatment, found to be associated with an unrecognized primary adenocarcinoma of the lung. Complete resolution of the eruption followed surgical resection of the tumor.


CorRelator: Interactive software for real-time high precision cryo-correlative light and electron microscopy.
Cryo-correlative light and electron microscopy (CLEM) is a technique that uses the spatiotemporal cues from fluorescence light microscopy (FLM) to investigate the high-resolution ultrastructure of biological samples by cryo-electron microscopy (cryo-EM). Cryo-CLEM provides advantages for identifying and distinguishing fluorescently labeled proteins, macromolecular complexes, and organelles from the cellular environment. Challenges remain on how correlation workflows and software tools are implemented on different microscope platforms to support automated cryo-EM data acquisition. Here, we present CorRelator: an open-source desktop application that bridges between cryo-FLM and real-time cryo-EM/ET automated data collection. CorRelator implements a pixel-coordinate-to-stage-position transformation for flexible, high accuracy on-the-fly and post-acquisition correlation. CorRelator can be integrated into cryo-CLEM workflows and easily adapted to standard fluorescence and transmission electron microscope (TEM) system configurations. CorRelator was benchmarked under live-cell and cryogenic conditions using several FLM and TEM instruments, demonstrating that CorRelator reliably supports real-time, automated correlative cryo-EM/ET acquisition, through a combination of software-aided and interactive alignment. CorRelator is a cross-platform software package featuring an intuitive Graphical User Interface (GUI) that guides the user through the correlation process. CorRelator source code is available at: https://github.com/wright-cemrc-projects/corr.


Cytotoxicities of sodium benzoate in primary culture of hepatocytes from adult rat liver.
The cytotoxicities of sodium benzoate was studied using primary culture of hepatocytes established from adult rat liver by a collagenase perfusion technique and maintained as a monolayer in serum-free culture medium. The activities of ornithine transcarbamylase (as a marker of mitochondria) and tyrosine aminotransferase (as a marker of cytosol) were clearly suppressed by sodium benzoate at concentration in excess of 500 micrograms/ml. Intracellular protein synthesis and DNA synthesis were also suppressed, and the suppression of DNA synthesis was observed even with a lower concentration of benzoate (100 micrograms/ml).


Molecular basis of cardiac hypertrophy.
Cardiac hypertrophy is an adaptive process to an increased hemodynamic overload. When cardiomyocytes cultured on silicone dishes were stretched, second messengers such as protein kinase C (PKC), Raf-1 kinase, and mitogen-activated protein (MAP) kinases were activated, which were followed by increased protein synthesis. Moreover, pretreatment with an angiotensin II (AngII) type 1 receptor antagonist dimished an increase in protein synthesis, MAP kinase activity, and c-fos gene expression induced by the stretching of cardiomyocytes. These suggest the linkage of the cardiac renin-angiotensin system to the formation of pressure-overload hypertrophy. Indeed, in the stretch-conditioned medium the levels of AngII concentration were increased. Also, mechanical stretch enhanced endothelin (ET)-1 release from the cardiomyocytes and activated the Na(+)/H(+) exchanger independently of these vasoactive peptides. In the second part, we examined AngII-induced signaling pathways both in cardiac myocytes and in cardiac fibroblasts. AngII-evoked signal transduction pathways differed between cell types. In cardiac fibroblasts AngII activated MAP kinases through a pathway including the Gbetagamma subunit of Gi protein, Src, Shc, Grb2, and Ras, while Gq and PKC activation was necessary in cardiac myocytes. We further explored norepinephrine (NE)-induced signaling pathways in cardiac myocytes. NE activated Raf-1 kinase and MAP kinases and increased amino acid uptake in cardiomyocytes of neonatal rats. beta-adrenoceptor (AR) stimulation as well and alpha1-AR stimulation was involved in NE-induced MAP kinase activation. It is noteworthy that unlike in other cell types not only PKC activation but also protein kinase A (PKA) activation increased the activities of Raf-1 kinase and MAP kinases in cardiac myocytes and induced cell growth. Finally, we observed that beta-AR-induced activation of MAP kinases is dependent on both Gs/cAMP/PKA and Gi/Src/Ras signaling pathways and that phosphorylation of beta-AR is critical to the cross talk between these signaling pathways.


Multiscale imaging and characterization of the effect of mixing temperature on asphalt concrete containing recycled components.
When producing asphalt concrete mixture with high amounts of reclaimed asphalt pavement (RAP), the mixing temperature plays a significant role in the resulting spatial distribution of the components as well as on the quality of the resulting mixture, in terms of workability during mixing and compaction as well as in service mechanical properties. Asphalt concrete containing 50% RAP was investigated at mixing temperatures of 140, 160 and 180°C, using a multiscale approach. At the microscale, using energy dispersive X-ray spectroscopy the RAP binder film thickness was visualized and measured. It was shown that at higher mixing temperatures this film thickness was reduced. The reduction in film thickness can be attributed to the loss of volatiles as well as the mixing of RAP binder with virgin binder at higher temperatures. X-ray computer tomography was used to characterize statistically the distribution of the RAP and virgin aggregates geometric features: volume, width and shape anisotropy. In addition using X-ray computer tomography, the packing and spatial distribution of the RAP and virgin aggregates was characterized using the nearest neighbour metric. It was shown that mixing temperature may have a positive effect on the spatial distribution of the aggregates but did not affect the packing. The study shows a tendency for the RAP aggregates to be more likely distributed in clusters at lower mixing temperatures. At higher temperatures, they were more homogeneously distributed. This indicates a higher degree of blending both at microscale (binder film) and macroscale (spatial distribution) between RAP and virgin aggregates as a result of increasing mixing temperatures and the ability to quantify this using various imaging techniques.


Detrusor underactivity is prevalent after radical prostatectomy: A urodynamic study including risk factors.
The objective was to determine the prevalence of, and factors that predict, detrusor underactivity (DU) in patients presenting with incontinence or lower urinary tract symptoms (LUTS) following radical prostatectomy (RP). We also determined the prevalence of bladder outlet obstruction (BOO) and detrusor overactivity (DO) in this population. Patients who underwent urodynamics post-RP were identified. Detrusor underactivity was defined as a maximum flow rate (Qmax) of ≤15 mL/s and detrusor pressure (Pdet) Qmax <20 cmH20 or maximum Pdet <20 cmH20 during attempted voiding. Abdominal voiding (AV) was defined as sustained increase in abdominal pressure during voiding. Bladder outlet obstruction and DO were identified using the Abrams-Griffiths nomogram and the International Continence Society criteria. Univariate logistic regression was used to determine factors predicting DU. The following factors were analyzed: age, year of RP, procedure type (minimally-invasive surgery [MIS] or open), postoperative radiation, nerve-sparing, clinical stage, biopsy Gleason grade and interval between RP and evaluation. Between 2005 and 2008, 264 patients underwent urodynamics post-RP. Detrusor underactivity was observed in 108 patients (41%; 95% CI 35%, 47%), of whom 48% demonstrated AV. Overall, BOO and DO were present in 17% (95% CI 12%, 22%) and 27% (95% CI 22%, 33%), respectively. On univariate analysis, only MIS RP was predictive of DU (univariate odds ratio 2.05 for MIS vs. open; p = 0.009). Detrusor underactivity and AV are common in patients presenting for evaluation of incontinence or LUTS following RP. The etiology of DU in this setting is likely related to the surgical approach. Because DU may affect the success of male incontinence treatment with the male sling or artificial urinary sphincter, it is useful to document its presence prior to treatment. More studies are needed to elucidate the influence of DU on treatment success for male urinary incontinence following RP.


Selecting a pharmacy layout design using a weighted scoring system.
A weighted scoring system was used to select a pharmacy layout redesign. Facilities layout design techniques were applied at a local hospital pharmacy using a step-by-step design process. The process involved observing and analyzing the current situation, observing the current available space, completing activity flow charts of the pharmacy processes, completing communication and material relationship charts to detail which areas in the pharmacy were related to one another and how they were related, researching applications in other pharmacies or in scholarly works that could be beneficial, numerically defining space requirements for areas within the pharmacy, measuring the available space within the pharmacy, developing a set of preliminary designs, and modifying preliminary designs so they were all acceptable to the pharmacy staff. To select a final layout that could be implemented in the pharmacy, those layouts were compared via a weighted scoring system. The weighted aspect further allowed additional emphasis on categories based on their effect on pharmacy performance. The results produced a beneficial layout design as determined through simulated models of the pharmacy operation that more effectively allocated and strategically located space to improve transportation distances and materials handling, employee utilization, and ergonomics. Facilities layout designs for a hospital pharmacy were evaluated using a weighted scoring system to identify a design that was superior to both the current layout and alternative layouts in terms of feasibility, cost, patient safety, employee safety, flexibility, robustness, transportation distance, employee utilization, objective adherence, maintainability, usability, and environmental impact.


The minimal important change for measures of balance and postural control in older adults: a systematic review.
The minimal important change and analogous terms (MIC) can provide a measure of change in health outcome variables that is associated with a level of importance for participant/patient. This review explores the availability of the MIC for different balance measures used with older adults in research and clinical settings. PubMed, ProQuest and Web of Science search engines were used and based on the inclusion and exclusion criteria, 11 studies were deemed suitable for data extraction and analysis. The results demonstrated that MIC is available for the following balance-associated tests: Berg Balance Scale, Timed Up and Go, Short Physical Performance Battery, BESTest and the Tinetti test. A range of MIC values were shown, reflective of different older adult health conditions, calculation methods and anchors used. It was also evident that the responsiveness of the test was not always available or appropriately determined, questioning the validity of the MIC value published. Greater research is needed to establish MIC for balance measurements for use with older adults with different health conditions, preferably using objective measures such as falls. The calculation of such statistics will improve the evaluation of intervention effectiveness.


The effect of volume ingested on rehydration and gastric emptying following exercise-induced dehydration.
The purpose of this study was to examine the effects of different drink volumes on rehydration, gastric emptying, and markers of fluid balance following exercise-induced dehydration. Nine male subjects (27.3 +/- 5.47 yr of age, 77.8 +/- 7.9 kg) exercised for 90 min (or until 2.5% of initial body weight was lost) on a cycle ergometer in a hot environment (30 degrees C with 60% RH). Following exercise, subjects were moved to a neutral environment (23 degrees C 50% RH) and rested for 30 min prior to beginning a 3-h rehydration period. During rehydration, subjects were serially fed with an electrolyte solution (14.98 mmol.l-1 Na+, 13.51 mmol.l-1 Cl-, and 7.95 mmol.l-1 K+) every 30 min with either 100% or 150% of the fluid lost during exercise. Gastric contents were determined every 15 min using double sampling. Blood samples, urine samples, and body weights were taken before and after exercise and at 1-h intervals throughout rehydration. Blood samples were analyzed for percent change in plasma volume, electrolyte concentration, aldosterone levels, and renin activity. Urine electrolyte concentrations were also measured. The final percent rehydration was 48.11 and 67.90 for the 100% and 150% conditions, respectively. During rehydration, the subjects emptied 98.9 and 86.0% of the fluid ingested, and the % emptied and used for weight gain at the end of rehydration was 55.1 and 54.6 for the 100% and 150% trials, respectively. Urine production was significantly higher in the 150 compared with the 100% condition while renin and aldosterone levels did not differ significantly.(ABSTRACT TRUNCATED AT 250 WORDS)


Costs of antibiotic resistance - separating trait effects and selective effects.
Antibiotic resistance can impair bacterial growth or competitive ability in the absence of antibiotics, frequently referred to as a 'cost' of resistance. Theory and experiments emphasize the importance of such effects for the distribution of resistance in pathogenic populations. However, recent work shows that costs of resistance are highly variable depending on environmental factors such as nutrient supply and population structure, as well as genetic factors including the mechanism of resistance and genetic background. Here, we suggest that such variation can be better understood by distinguishing between the effects of resistance mechanisms on individual traits such as growth rate or yield ('trait effects') and effects on genotype frequencies over time ('selective effects'). We first give a brief overview of the biological basis of costs of resistance and how trait effects may translate to selective effects in different environmental conditions. We then review empirical evidence of genetic and environmental variation of both types of effects and how such variation may be understood by combining molecular microbiological information with concepts from evolution and ecology. Ultimately, disentangling different types of costs may permit the identification of interventions that maximize the cost of resistance and therefore accelerate its decline.


Endometrial carcinoma with recurrence in the incisional scar: a case report.
We report a case of papillary adenocarcinoma of uterus which developed a recurrence over the scar of surgery. The patient initially underwent surgery followed by adjuvant chemoradiotherapy for her stage II disease. She was disease-free until 21 months when she developed a small mass over the lower site of incisional scar. Fine needle aspiration cytology from this swelling revealed metastatic papillary adenocarcinoma similar to previous histopathology. Treatment of the scar recurrence consisted of palliative radiation therapy and chemotherapy.


Modeling the apparent frequency-specific suppression in simple cell responses.
Simple cells in cat striate cortex are selective for spatial frequency. It is widely believed that this selectivity arises simply because of the way in which the neurons sum inputs from the lateral geniculate nucleus. Alternate models, however, advocate the need for frequency-specific inhibitory mechanisms to refine the spatial frequency selectivity. Indeed, simple cell responses are often suppressed by superimposing stimuli with spatial frequencies that flank the neuron's preferred spatial frequency. In this article, we compare two models of simple cell responses head-to-head. One of these models, the flanking-suppression model, includes an inhibitory mechanism that is specific to frequencies that flank the neuron's preferred spatial frequency. The other model, the nonspecific-suppression model, includes a suppressive mechanism that is very broadly tuned for spatial frequency. Both models also include a rectification nonlinearity and both may include an additional accelerating (e.g., squaring) output nonlinearity. We demonstrate that both models can be consistent with the apparent flanking suppression. However, based on other experimental results, we argue that the nonspecific-suppression model is more plausible. We conclude that the suppression is probably broadly tuned for spatial frequency and that the apparent flanking suppression is actually due to distortions introduced by an accelerating output nonlinearity.


Trends in vegetation productivity related to climate change in China's Pearl River Delta.
Climate change will be a powerful stressor on ecosystems and biodiversity in the second half of the 21st century. In this study, we used the satellite-derived Normalized Difference Vegetation Index (NDVI) to examine a 34-year trend along with the response of vegetation to climate indicators surrounding the world's largest megacity: the Pearl River Delta (PRD) of China. An overall increasing trend is observed in vegetation productivity metrics over the study period 1982 to 2015. Increase in winter productivity in both natural ecosystems and croplands is more related to increasing temperatures (r = 0.5-0.78), than to changes in rainfall. For growing season productivity, negative correlations with temperature were observed in cropland regions, and some forests in the northern part of PRD region, suggesting high-temperature stress on crop production and forest vegetation. However, increased winter and spring temperatures provide higher opportunities for cropping in winter. During the decade 1995-2004, vegetation productivity metrics showed a reversal in the upward trend. The geographical and biological complexity of the region under significant climatic and development impacts suggests causative factors would be synergistic. These include our observed decrease in sunshine hours, increasing cloud cover associated with atmospheric aerosols from industrial and urban development, direct pollution effects on plant growth, and exceedance of high temperature growth thresholds.


Development of a robust UPLC-MS/MS method for the quantification of riluzole in human plasma and its application in pharmacokinetics.
Introduction: The aim of the present study was to establish a simple method for the determination of riluzole in human plasma by ultraperformance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) and apply it for the determination of riluzole in amyotrophic lateral sclerosis (ALS) patients. Methods: Samples were prepared by protein precipitation and were then gradient-eluted on a column of ACQUITY UPLC® HSS T3 by using 0.1% formic acid acetonitrile and 0.1% formic acid water as the mobile phase. Detection was performed on a Xevo TQ-S tandem mass spectrometer in the multiple-reaction monitoring mode using positive electrospray ionization. Validation was performed in the range of 5-800 ng/mL. Results and discussion: Three batches of precision accuracy, selectivity, matrix effects, extraction recovery, and stability were also verified and met the requirements. The results showed that the method was reliable and successfully applied to the pharmacokinetics study of riluzole in Chinese amyotrophic lateral sclerosis patients. Meanwhile, in comparison with other prior published methods, our method has the advantages of simple sample preparation, relatively short running time, and small plasma sample consumption, which represented a high-throughput sample determination potential.


Functional role of glial-derived neurotrophic factor in a mixed allergen murine model of asthma.
Our previous study showed that glial-derived neurotrophic factor (GDNF) expression is upregulated in asthmatic human lungs, and GDNF regulates calcium responses through its receptor GDNF family receptor α1 (GFRα1) and RET receptor in human airway smooth muscle (ASM) cells. In this study, we tested the hypothesis that airway GDNF contributes to airway hyperreactivity (AHR) and remodeling using a mixed allergen mouse model. Adult C57BL/6J mice were intranasally exposed to mixed allergens (ovalbumin, Aspergillus, Alternaria, house dust mite) over 4 wk with concurrent exposure to recombinant GDNF, or extracellular GDNF chelator GFRα1-Fc. Airway resistance and compliance to methacholine were assessed using FlexiVent. Lung expression of GDNF, GFRα1, RET, collagen, and fibronectin was examined by RT-PCR and histology staining. Allergen exposure increased GDNF expression in bronchial airways including ASM and epithelium. Laser capture microdissection of the ASM layer showed increased mRNA for GDNF, GFRα1, and RET in allergen-treated mice. Allergen exposure increased protein expression of GDNF and RET, but not GFRα1, in ASM. Intranasal administration of GDNF enhanced baseline responses to methacholine but did not consistently potentiate allergen effects. GDNF also induced airway thickening, and collagen deposition in bronchial airways. Chelation of GDNF by GFRα1-Fc attenuated allergen-induced AHR and particularly remodeling. These data suggest that locally produced GDNF, potentially derived from epithelium and/or ASM, contributes to AHR and remodeling relevant to asthma.NEW & NOTEWORTHY Local production of growth factors within the airway with autocrine/paracrine effects can promote features of asthma. Here, we show that glial-derived neurotrophic factor (GDNF) is a procontractile and proremodeling factor that contributes to allergen-induced airway hyperreactivity and tissue remodeling in a mouse model of asthma. Blocking GDNF signaling attenuates allergen-induced airway hyperreactivity and remodeling, suggesting a novel approach to alleviating structural and functional changes in the asthmatic airway.


Experience of prophylactic thyroidectomy in multiple endocrine neoplasia type 2A kindreds with RET codon 804 mutations.
Genetic screening in multiple endocrine neoplasia type 2 (MEN 2) has led to specific management guidelines based on genotype-phenotype analysis. However, there is controversy regarding the appropriate age for prophylactic thyroidectomy in families with mutations in codon 804 in exon 14 of the RET proto-oncogene, where medullary thyroid cancer (MTC) may not develop until adulthood. We prospectively studied two MEN 2A families, one with the V804L and the other with the V804M RET mutation, to report our experience of genetic and biochemical screening and prophylactic thyroidectomy. Family 1 is one of the largest MEN 2A families in the literature, where 22 prophylactic thyroidectomies have been performed. C-cell hyperplasia (CCH) was found in 23 out of 25 thyroidectomy specimens from family members of ages 5 years and upwards. MTC was found in 10 out of 18 adults of age 25 years upwards, including the family 2 proband, who was found to have MTC with lymph node metastases at age 28. Phaeochromocytoma was only observed in one patient, but six cases of histologically confirmed hyperparathyroidism were seen in family 1. We suggest that prophylactic thyroidectomy should not be delayed until adulthood in MEN 2A families carrying codon 804 RET mutations, but should be performed when there is CCH, before the development of MTC, as close as possible to age 6 years, which is the age of the youngest reported case of MTC in '804' families.


Using fMRI to study reward processing in humans: past, present, and future.
Functional magnetic resonance imaging (fMRI) is a noninvasive tool used to probe cognitive and affective processes. Although fMRI provides indirect measures of neural activity, the advent of fMRI has allowed for1) the corroboration of significant animal findings in the human brain, and2) the expansion of models to include more common human attributes that inform behavior. In this review, we briefly consider the neural basis of the blood oxygenation level dependent signal to set up a discussion of how fMRI studies have applied it in examining cognitive models in humans and the promise of using fMRI to advance such models. Specifically, we illustrate the contribution that fMRI has made to the study of reward processing, focusing on the role of the striatum in encoding reward-related learning signals that drive anticipatory and consummatory behaviors. For instance, we discuss how fMRI can be used to link neural signals (e.g., striatal responses to rewards) to individual differences in behavior and traits. While this functional segregation approach has been constructive to our understanding of reward-related functions, many fMRI studies have also benefitted from a functional integration approach that takes into account how interconnected regions (e.g., corticostriatal circuits) contribute to reward processing. We contend that future work using fMRI will profit from using a multimodal approach, such as combining fMRI with noninvasive brain stimulation tools (e.g., transcranial electrical stimulation), that can identify causal mechanisms underlying reward processing. Consequently, advancements in implementing fMRI will promise new translational opportunities to inform our understanding of psychopathologies.


Including RNA secondary structures improves accuracy and robustness in reconstruction of phylogenetic trees.
In several studies, secondary structures of ribosomal genes have been used to improve the quality of phylogenetic reconstructions. An extensive evaluation of the benefits of secondary structure, however, is lacking. This is the first study to counter this deficiency. We inspected the accuracy and robustness of phylogenetics with individual secondary structures by simulation experiments for artificial tree topologies with up to 18 taxa and for divergency levels in the range of typical phylogenetic studies. We chose the internal transcribed spacer 2 of the ribosomal cistron as an exemplary marker region. Simulation integrated the coevolution process of sequences with secondary structures. Additionally, the phylogenetic power of marker size duplication was investigated and compared with sequence and sequence-structure reconstruction methods. The results clearly show that accuracy and robustness of Neighbor Joining trees are largely improved by structural information in contrast to sequence only data, whereas a doubled marker size only accounts for robustness. Individual secondary structures of ribosomal RNA sequences provide a valuable gain of information content that is useful for phylogenetics. Thus, the usage of ITS2 sequence together with secondary structure for taxonomic inferences is recommended. Other reconstruction methods as maximum likelihood, bayesian inference or maximum parsimony may equally profit from secondary structure inclusion. This article was reviewed by Shamil Sunyaev, Andrea Tanzer (nominated by Frank Eisenhaber) and Eugene V. Koonin. Reviewed by Shamil Sunyaev, Andrea Tanzer (nominated by Frank Eisenhaber) and Eugene V. Koonin. For the full reviews, please go to the Reviewers' comments section.


Single-cell electroporation arrays with real-time monitoring and feedback control.
Rapid well-controlled intracellular delivery of drug compounds, RNA, or DNA into a cell--without permanent damage to the cell--is a pervasive challenge in basic cell biology research, drug discovery, and gene delivery. To address this challenge, we have developed a bench-top system comprised of a control interface, that mates to disposable 96-well-formatted microfluidic devices, enabling the individual manipulation, electroporation and real-time monitoring of each cell in suspension. This is the first demonstrated real-time feedback-controlled electroporation of an array of single-cells. Our computer program automatically detects electroporation events and subsequently releases the electric field, precluding continued field-induced damage of the cell, to allow for membrane resealing. Using this novel set-up, we demonstrate the reliable electroporation of an array (n = 15) of individual cells in suspension, using low applied electric fields (<1 V) and the rapid and localized intracellular delivery of otherwise impermeable compounds (Calcein and Orange Green Dextran). Such multiplexed electrical and optical measurements as a function of time are not attainable with typical electroporation setups. This system, which mounts on an inverted microscope, obviates many issues typically associated with prototypical microfluidic chip setups and, more importantly, offers well-controlled and reproducible parallel pressure and electrical application to individual cells for repeatability.


Theoretical and experimental study on the Nd:YAG/BaWO4/KTP yellow laser generating 8.3 W output power.
A diode-side-pumped actively Q-switched intracavity frequency-doubled Nd:YAG/BaWO(4)/KTP Raman laser is studied experimentally and theoretically. Rate equations are used to analyze the Q-switched yellow laser by considering the transversal distributions of the intracavity photon density and the inversion population density. An 8.3 W 590 nm laser is obtained with a 125.8 W 808 nm pump power and a 15 kHz pulse repetition frequency. The corresponding optical conversion efficiency from diode laser to yellow laser is 6.57%, much higher than that of the former reported side-pumped yellow laser. The output powers with respect to the incident pump power are in agreement with the theoretical results on the whole.


A 5-day Heat Acclimation Program Improves Heat Stress Indicators While Maintaining Exercise Capacity.
Wardenaar, FC, Ortega-Santos, CP, Vento, K, Beaumont, JS, Griffin, SC, Johnston, C, and Kavouras, SA. A 5-day heat acclimation program improves heat stress indicators while maintaining exercise capacity. J Strength Cond Res 35(5): 1279-1286, 2021-This study aimed to evaluate whether a daily 60 minutes isothermic biking protocol during a 5-day period could improve physiological heat acclimation and exercise performance capacity in partially acclimated subjects. A quasi-experimental study consisted of an intervention (INT, n = 7) and control (CON, n = 7) group completing 2 12 minutes Cooper tests (pre-CT on day 1 and post-CT on day 7) and a heat stress test (HST, on day 9). INT performed additional intensive exercise 1 hour per day on days 1-5, whereas CON did not. During CTs and HST, core temperature (Tc, telemetric capsule), skin temperature (Tsk, sensors at neck, right shoulder, left hand, and right shin), and heart rate (HR, chest strap) were continuously monitored and baseline, average, peak, and increment were calculated. During the HST, the INT group showed a smaller baseline-peak Tc increment (INT 0.88 ± 0.27 vs. CON 1.64 ± 0.90° C, p = 0.02), a lower HR peak (150.2 ± 12.6 vs. 173.0 ± 16.8 b·min-1, p = 0.02), and lower Tsk peak (36.47 ± 0.62 vs. 36.54 ± 0.46° C, p = 0.04). There was a nonsignificant, but practical difference based on a moderate effect size for change in pre-CT to post-CT performance of nearly +2.7 ± 12.3% in INT and -3.0 ± 8.5% in CON (p = 0.32 and d = 0.51), and HST distance covered resulting in a nonsignificant difference of 464 ± 849 m between INT and CON (p = 0.38 and d = 0.44). In conclusion a short-term 5-day heat acclimation program including 300 minutes of extra exercise resulted in positive physiological adaptions to heat stress, as indicated by lower core temperature and HR in comparison with a control group.


Links between Adolescent Athletes' Prosocial Behavior and Relationship with Parents: A Mixed Methods Study.
Adolescent relationships with parents are of the highest importance. The relationships likely reflect the nature of internal working models in youth sport that may well function as a psychological template during the construction of youth prosocial behavior. However, researchers' focus to date has concerned specific aspects of parental practices in child-based sporting activities. There is a lack of research covering parent-athlete interpersonal relationships concerned with how the relationships affect adolescent prosocial behavior. The purpose of this mixed methods explanatory sequential study was to examine teenage athletes' prosocial behavior and their relationships with parents. To achieve our purpose, we obtained quantitative data from 1348 athletes and non-athletes (ages 12⁻16), and qualitative data from 12 adolescent athletes and 12 youth sports parents. In the quantitative phase, we assessed adolescent prosocial behavior regarding the following six forms of prosocial behavior: public, anonymous, dire, compliant, altruistic and emotional. In the qualitative follow-up, three themes emerged from the adolescent athlete's perspective: (1) sport as an escape; (2) parent-child relationships in youth sports; (3) adolescents' desired behavior. Three themes emerged from the parental perspective: (1) sport as protection and as a school of life; (2) painful decisions to release a child; (3) understanding adolescent behavior. We found protection from delinquent behavior and increased prosocial behavior with securely attached young athletes who are actively involved in sports.


The relationships between pain, disability, and health-related quality of life: an 8-year follow-up study of female home care personnel.
To investigate the development of pain conditions, disability, and health-related quality of life over an 8-year period in home care personnel. In earlier studies of 607 women, we reported baseline data concerning home care personnel. This study reports the results from an 8-year follow-up using a postal questionnaire. The questionnaire was completed by 87%. Prevalences of pain in upper back, lower back, and knees as well as pain intensity of the low back had decreased. Participants with the highest pain intensities of the low back at baseline had relatively lower pain intensities at follow-up. Anatomical spreading of pain was associated with higher average pain intensity. Disability had increased significantly during the time period. In the regression of disability at follow-up, average pain intensity together with disability rating index at baseline were the most important regressors; a similar pattern was found for quality of life. The development of disability differed from that of low-back pain intensity. Spreading of pain and pain intensity across the anatomical regions influenced disability and quality of life over 8 years. When assessing pain, it seems important to determine the spread of pain rather than just focusing on the area with intense pain. Spreading of pain and the average intensity of pain across the involved anatomical regions have importance for future pain and disability and quality of life. The clinical assessment of subjects with chronic pain prior to rehabilitation interventions has to determine the spreading of pain rather than just focusing on the area with the most intense pain. The different developments over time for pain intensity and disability indicate the need for applying a bio-psycho-social view of pain both when assessing the patient with pain and when discussing the prognosis and course of the actual pain condition with the patient.


Factors relevant in the reaction of pyrroloquinoline quinone with amino acids. Analytical and mechanistic implications.
In order to reveal the stability of pyrroloquinoline quinone (PQQ) in complex samples, its reaction on incubation with amino acids was followed spectrophotometrically by monitoring oxygen consumption, and with a biological assay. For several alpha-amino acids, the formation of a yellow coloured compound (lambda max = 420 nm) was accompanied by oxygen uptake and disappearance of biological activity from the reaction mixture. The yellow product appeared to be an oxazole of PQQ, the exact structure depending on the amino acid used. Oxazole formation also occurred under anaerobic conditions with concomitant formation of PQQH2, suggesting that PQQ is able to oxidize the presumed oxazoline to the oxazole. Besides the condensation reaction, there is also a catalytic cycle in which an aldimine adduct of PQQ and the amino acid is converted into the aminophenol form of the cofactor and an aldehyde resulting from oxidative decarboxylation of the amino acid. Addition of NH4+ salts, as well as that of certain divalent cations, greatly stimulated both the cyclic and the linear reaction. With basic amino acids, oxazole formation scarcely occurred. However, as oxygen consumption was observed (provided that certain divalent cations were present), conversion of these compounds took place. A reaction scheme is proposed accounting for the products formed and the effects observed. Since NH4+ ions activate several quinoproteins (PQQ-containing enzymes) and divalent cations (Ca2+, Fe2+, and Cu2+) are additional (co)factors in certain metallo quinoproteins, the effects of metal ions observed here could be related to the mechanistic features of these enzymes. Although all oxazoles were converted to PQQ by acid hydrolysis, PQQ was not detected when hydrolysis was carried out in the presence of tryptophan, a compound which appeared to have a deleterious effect on the cofactor under this condition. The results here described explain why analysis methods for free PQQ in complex samples fail in certain cases, or are not quantitative.


[Routine medical management of obesity].
Management of obesity is often reduced to counselling, focusing on factors directly affecting energy balance: food intake and physical activity. However, the causes of excessive food intake, eating disorders and their causal factors, are largely unknown. This can lead to unsuitable obesity management. In the present text, we detail the clinical elements which can be helpful for the specialist or general practitioner making a global assessment of their overweight patients. The goal is to help the patient establish a care project with medical support, as would be the case for any chronic disease.


Mucilaginibacter frigoritolerans sp. nov., Mucilaginibacter lappiensis sp. nov. and Mucilaginibacter mallensis sp. nov., isolated from soil and lichen samples.
Five cold-adapted bacteria belonging to the genus Mucilaginibacter were isolated from lichen and soil samples collected from Finnish Lapland and investigated in detail by phenotypic and phylogenetic analyses. Based on 16S rRNA gene phylogeny, the novel strains represent three new branches within the genus Mucilaginibacter. The strains were aerobic, chemo-organotrophic, non-motile rods and formed pigmented, smooth, mucoid colonies on solid media. The strains grew between 0 and 33 °C (optimum growth at 25 °C) and at pH 4.5-8.0 (optimum growth at pH 6.0). The main cellular fatty acids were iso-C(15 : 0), summed feature 3 (C(16 : 1)ω7c/iso-C(15 : 0) 2-OH) and iso-C(17 : 0) 3-OH and the major respiratory quinone was MK-7. The DNA G+C contents were 44.0-46.5 mol%. Based on phylogenetic, phenotypic and chemotaxonomic data, the strains represent three novel species of the genus Mucilaginibacter for which the names Mucilaginibacter frigoritolerans sp. nov. (type strain FT22(T) =ATCC BAA-1854(T) =LMG 25359(T)), Mucilaginibacter lappiensis sp. nov. (type strain ANJLI2(T) =ATCC BAA-1855(T) =LMG 25358(T)) and Mucilaginibacter mallensis sp. nov. (type strain MP1X4(T) =ATCC BAA-1856(T) =LMG 25360(T)) are proposed.


[Formation of flavanolignane in the white-flowered variety (Szibilla) of Silybum marianum in relation to fruit development].
In our present work we studied the white-flowered lady's thistle variety 'Szibilla' improved by our Institute and having received state acknowledgement in 1997. During our investigations we established the following facts: --flavanolignanes having liver-protecting, cytoprotective effects can be found only in the developing and ripe achene fruits. The vegetative organs contain flavanoids of different types; --around the tissues of the fruit (Silybi mariani fructus) known as drug raw material for therapy and the pharmaceutical industry, effective agents are contained only in the fruit wall having undergone significant changes as well as in the woody (ligneous) seed-coat. The endosperm and the plant embryo contain effective agents only in traces; --during ontogenesis--following the pistil's organization into fruit and seed, respectively--we revealed that the biogenesis and accummulation of flavanolignanes correlate with the wood formation (lignification) of the seed-coat.


Head shape and bite performance in xenosaurid lizards.
Bite performance in lizards influences many aspects of the animal's lifestyle. During feeding, inter- and intrasexual interactions, and defensive behavior, the ability to bite hard might be advantageous. Although biomechanical considerations predict clear relations between head shape and bite performance, this has rarely been tested. Here we investigate the effect of head shape on bite performance in three closely related species of xenosaurid lizards. Our data show that in this family of lizards, bite performance is mainly determined by head height, with high headed animals biting harder than flat headed ones. Species clearly differ in head shape and bite performance and show a marked sexual dimorphism. The dimorphism in head shape also results in an intersexual difference in bite performance. As head height is the major determinant of bite performance in xenosaurid lizards, trade offs between a crevice dwelling life-style and bite performance seem to occur. The evolutionary implications of these results are discussed. J. Exp. Zool. 290:101-107, 2001.


[Autoradiographic study of the ontogeny of the imipramine receptors in the cerebral cortex of rats].
Localization of 3H-imipramine binding sites in the cerebral cortex of rats was studied on the 19th day of prenatal development and on the 3d and 14th days of postnatal ontogenesis. Imipramine was injected into rats on days 17, 18 and 19 of gestation. The maximal imipramine receptor density was found in cortex layers IV-V. The therapeutic doses of imipramine provoked marked stimulation of the formation of imipramine receptors on the 3d day of postnatal development. However, on the 14th day the 3H-imipramine binding level was stabilized.


Nasal Herpes Simplex With Infraorbital Neuralgia: A Rare Presentation.
Facial herpes is a form of herpes simplex type I infection and presents with characteristic vesicular lesion around the perioral region. Nasal herpes, a form of facial herpes is a rare presentation with only a few cases reported in the literature. Neuralgic pain in herpes simplex is usually experienced at the site of the lesion during or before the eruptive stage. Here, we are reporting a case where the patient with a herpes simplex lesion over the tip of her nose presented with pain over the infraorbital region, which is a region supplied by the infraorbital nerve, a branch of the maxillary division of the trigeminal nerve. Initially confused as a bacterial infection due to its unusual presentation and rarity of the condition, the patient was given anti-bacterial therapy, but on showing no relief in symptoms, the patient was treated with appropriate antiviral drugs, following which complete remission of the lesion was observed. The case highlights a rare site for a common condition and atypical presentation.


Comparison of puberty and psychosocial adjustment between Taiwanese adolescent females with and without diabetes.
The aims of this study were to (1) determine the differences in puberty and psychosocial adjustment among Taiwanese adolescent females with and without type 1 diabetes mellitus, and (2) examine the interaction between pubertal timing and diabetes in relation to its effect on adolescent's psychosocial adjustment. Rapid physical and sexual changes during puberty elicit a wide array of psychosocial adjustments. The effects of pubertal changes among adolescent females with type 1 diabetes mellitus on psychosocial adjustment are unknown. Design. This study used a comparative, case-controlled design. A total of 82 adolescent females, aged 10-17, were recruited for the study. Forty-one adolescents with type 1 diabetes mellitus were age-matched to 41 adolescents without type 1 diabetes mellitus. Adolescents' psychosocial adjustment, including internalising and externalising behaviours, was assessed using the Child Behaviour Checklist (parental report) and the Youth Self-Report (individual self-report). The self-reported Pubertal Development Scale was used to measure adolescents' puberty, including onset of menses, age at menarche and pubertal timing. When compared to their counterparts, adolescent females with type 1 diabetes mellitus reported a delayed menarche and a delayed puberty. Females with type 1 diabetes mellitus had significantly greater internalising and externalising behaviours than their counterparts according to parental reports. Onset of menses and adolescent self-reported psychosocial adjustment were not different between the two groups. Interaction analyses showed that the association between pubertal timing and internalising behaviours was related to the presence of diabetes. According to parental reports, early pubertal timing had positive effects on internalising behaviours for adolescent females with type 1 diabetes mellitus not for adolescent females without type 1 diabetes mellitus. A multi-informant approach is suggested when health care professionals assess adolescent psychosocial adjustment. Health care professionals must provide female teenagers with information and opportunities to discuss the impact of type 1 diabetes mellitus on their puberty and psychosocial adjustment.


Binding of proteolytically-degraded human colonic mucin glycoproteins to the Gal/GalNAc adherence lectin of Entamoeba histolytica.
Rat and human colonic mucin glycoproteins bind to the Gal/GalNAc adherence lectin on the surface of Entamoeba histolytica in vitro, thus inhibiting the organism from adhering to and lysing the target cells. Human colonic mucin glycoproteins were isolated by Sepharose 4B gel filtration chromatography, they were proteolytically degraded with trypsin, pronase, and papain, and the glycoprotein fractions were reisolated by Sephacryl S-200 gel filtration chromatography. Binding of the mucin glycoprotein fractions to amoebae was quantitated by the inhibition of adherence of Chinese hamster ovary cells to the surface of the amoebae. Trypsin and papain digests caused 40 and 20% reductions, respectively, in the excluded fractions (void volume) that contained all the carbohydrates; pronase digests resulted in extensive degradation of the mucin glycoprotein with the carbohydrate fractions eluting over 40% of the gel bed volume. 3H-labelled mucin glycoprotein and sodium dodecylsulfate-polyacrylamide gel electrophoresis confirmed the presence of the high molecular weight carbohydrate-rich glycoproteins with no subunits in the excluded fractions and the absence of sugars in the included peptides. Only the high molecular weight carbohydrate-containing fractions bind amoebae and inhibit amoebic adherence to Chinese hamster ovary cells. The trypsin digested mucins in the excluded volume were more efficient than the native undigested mucins in binding amoebae. The carbohydrate-containing fractions of the pronase digests were the least effective in binding amoebae and inhibiting adherence of Chinese hamster ovary cells. This suggests that proteolytically-degraded colonic mucins that are glycosylated, as well as the undegraded native mucin glycoproteins of the gut, may play a protective role in binding to amoebae, thus preventing contact of amoebae with mucosal epithelial cells and potential invasion.


Global variations in health: evaluating Wilkinson's income inequality hypothesis using the World Values Survey.
This international comparative study analyses individual-level data derived from the World Values Survey to evaluate Wilkinson's [(1996). Unhealthy societies: The afflictions of inequality. London: Routledge; (1998). Mortality and distribution of income. Low relative income affects mortality [letter; comment]. British Medical Journal, 316, 1611-1612] income inequality hypothesis regarding variations in health status. Random-coefficient, multilevel modelling provides a direct test of Wilkinson's hypothesis using micro-data on individuals and macro-data on income inequalities analysed simultaneously. This overcomes the ecological fallacy that has troubled previous research into links between individual self-rated health, individual income, country income and income inequality data. Logic regression analysis reveals that there are substantial differences between countries in self-rated health after taking account of age and gender, and individual income has a clear effect in that poorer people report experiencing worse health. The Wilkinson hypothesis is not supported, however, since there is no significant relationship between health and income inequality when individual factors are taken into account. Substantial differences between countries remain even after taking account of micro- and macro-variables; in particular the former communist countries report high levels of poor health.


Modes of cognition: Evidence from metastable brain dynamics.
Managing cognitive load depends on adequate resource allocation by the human brain through the engagement of metastable substates, which are large-scale functional networks that change over time. We employed a novel analysis method, deep autoencoder dynamical analysis (DADA), with 100 healthy adults selected from the Human Connectome Project (HCP) data set in rest and six cognitive tasks. The deep autoencoder of DADA described seven recurrent stochastic metastable substates from the functional connectome of BOLD phase coherence matrices. These substates were significantly differentiated in terms of their probability of appearance, time duration, and spatial attributes. We found that during different cognitive tasks, there was a higher probability of having more connected substates dominated by a high degree of connectivity in the thalamus. In addition, compared with those during tasks, resting brain dynamics have a lower level of predictability, indicating a more uniform distribution of metastability between substates, quantified by higher entropy. These novel findings provide empirical evidence for the philosophically motivated cognitive theory, suggesting on-line and off-line as two fundamentally distinct modes of cognition. On-line cognition refers to task-dependent engagement with the sensory input, while off-line cognition is a slower, environmentally detached mode engaged with decision and planning. Overall, the DADA framework provides a bridge between neuroscience and cognitive theory that can be further explored in the future.


Creatine ethyl ester: a new substrate for creatine kinase.
The creatine kinase/phosphocreatine system plays a key role in cell energy buffering and transport, particularly in cells with high or fluctuating energy requirements, like neurons, i.e. it participates in the energetic metabolism of the brain. Creatine depletion causes several nervous system diseases, alleviated by phosphagen supplementation. Often, the supplementation contains both creatine and creatine ethyl ester, known to improve the effect of creatine through an unknown mechanism. In this work we showed that purified creatine kinase is able to phosphorilate the creatine ethyl ester. The K(m) and V(max) values, as well as temperature and pH optima were determined. Conversion of the creatine ethyl ester into its phosphorylated derivative, sheds light on the role of the creatine ethyl ester as an energy source in supplementation for selected individuals.


Association of fibroblast growth factor receptor-1 with c-Src correlates with association between c-Src and cortactin.
The initiation of maximal DNA synthesis by fibroblast growth factor (FGF)-1 requires the presence of the growth factor during the entire G0 to G1 transition period of the cell cycle (Zhan, X., Hu, X., Friesel, R., and Maciag, T. (1993) J. Biol. Chem. 268, 9611-9620). During this time, the phosphorylation of several novel proteins on tyrosine residues occurs, and one of these phosphotyrosyl-containing proteins has been characterized as the murine homolog of the chicken cortactin gene (Zhan, X., Hu, X., Hampton, B., Burgess, W.H., Friesel, R., and Maciag, T. (1993) J. Biol. Chem. 268, 24427-24431), a putative substrate for v-Src. We have examined the possibility that FGF-1 employs c-Src or Src-like kinases as signaling intermediates during the mid and late G1 phase of the NIH 3T3 cell cycle using immunoprecipitation and immunoblot analysis. We have demonstrated that c-Src can associate with cortactin in a FGF-1-dependent manner. We have also demonstrated that a monoclonal antibody prepared against FGF receptor (R)-1 is able to co-precipitate Src-related proteins in lysates from FGF-1-treated NIH 3T3 cells. Furthermore, a kinase-active form of FGFR-1 expressed in a bacterial system was also able to associate with Src kinases in a manner dependent on the phosphorylation status of the FGFR-1 protein. Lastly, the Src homology (SH)-2 domain of v-Src was able to recognize a recombinant form of FGFR-1. Because (i) the association between FGFR-1 and Src-like kinases exhibits kinetics similar to those observed between the Src kinases and cortactin and (ii) the Src-SH2 domain is likely to be involved in the association with FGFR-1, we propose that the association of c-Src with activated FGF receptors may be responsible for the tyrosine phosphorylation of cortactin during the mid to late G1 phase of the cell cycle.


Splicing Factor SRSF1 Promotes Pancreatitis and KRASG12D-Mediated Pancreatic Cancer.
Inflammation is strongly associated with pancreatic ductal adenocarcinoma (PDAC), a highly lethal malignancy. Dysregulated RNA splicing factors have been widely reported in tumorigenesis, but their involvement in pancreatitis and PDAC is not well understood. Here, we report that the splicing factor SRSF1 is highly expressed in pancreatitis, PDAC precursor lesions, and tumors. Increased SRSF1 is sufficient to induce pancreatitis and accelerate KRASG12D-mediated PDAC. Mechanistically, SRSF1 activates MAPK signaling-partly by upregulating interleukin 1 receptor type 1 (IL1R1) through alternative-splicing-regulated mRNA stability. Additionally, SRSF1 protein is destabilized through a negative feedback mechanism in phenotypically normal epithelial cells expressing KRASG12D in mouse pancreas and in pancreas organoids acutely expressing KRASG12D, buffering MAPK signaling and maintaining pancreas cell homeostasis. This negative feedback regulation of SRSF1 is overcome by hyperactive MYC, facilitating PDAC tumorigenesis. Our findings implicate SRSF1 in the etiology of pancreatitis and PDAC, and point to SRSF1-misregulated alternative splicing as a potential therapeutic target. We describe the regulation of splicing factor SRSF1 expression in the context of pancreas cell identity, plasticity, and inflammation. SRSF1 protein downregulation is involved in a negative feedback cellular response to KRASG12D expression, contributing to pancreas cell homeostasis. Conversely, upregulated SRSF1 promotes pancreatitis and accelerates KRASG12D-mediated tumorigenesis through enhanced IL1 and MAPK signaling. This article is highlighted in the In This Issue feature, p. 1501.


The group-as-a-whole-object relations model of group psychotherapy.
The authors review the theoretical basis of group psychotherapy performed at The Menninger Clinic and demonstrate how the theory has been put into practice on two different types of inpatient units. The fundamental elements of the theory and practice used can be traced to object relations theory as originally proposed by Melanie Klein. Her work with individuals was directly applied to working with groups by Ezriel and Bion, who focused on interpreting group tension. More modern approaches have reintegrated working with individual concerns while also attending to the group-as-a-whole. Historically, these principles have been applied to long-term group treatment. The authors apply the concepts from the group-as-a-whole literature to short- and medium-length inpatient groups with open membership. They offer clinical examples of the application of these principles in short-term inpatient settings in groups with open membership.


Single nucleotide polymorphism analysis of corticotropin-releasing factor-binding protein gene in bipolar disorder.
Corticotropin-releasing factor-binding protein regulates the availability of free corticotropin-releasing factor and is a functional candidate gene for affective disorders. The aim of this study was to examine the association between polymorphisms in CRF-BP gene and bipolar disorder in an isolated Swedish population. One hundred and eighty-two patients with bipolar I disorder and 333 controls from Northern Sweden were included in the study. Five single nucleotide polymorphisms and a deletion polymorphism in the CRF-BP gene were genotyped. The haplotype block structure of the gene was considered and the expectation maximization algorithm was adopted to estimate the haplotype frequencies. As a result, there were no significant associations of the different polymorphisms in the CRF-BP gene with bipolar disorder. In conclusion, this study in an isolated Swedish population does not support a role for the CRF-BP gene in the vulnerability for bipolar disorder.


Towards New Broader Spectrum Pneumococcal Vaccines: The Future of Pneumococcal Disease Prevention.
Seven-valent pneumococcal conjugate vaccine (PCV7) introduction and routine pediatric use has substantially reduced the burden of Streptococcus pneumoniae disease worldwide. However, a significant amount of disease burden, due to serotypes not contained in PCV7, still exists globally. A newly recognized serotype, 6C, was until recently, identified and reported as serotype 6A. This review summarizes the serotype epidemiology of pneumococcal disease pre- and post-introduction of PCV7, available post-marketing surveillance data following the introduction of higher valency pneumococcal vaccines (PCV10, PCV13) and future prospects for the development of new pneumococcal vaccines.


[Giant hemangioma treated with interferon alpha-2a].
The authors describe a case of giant hemangioma treated with interferon alpha-2a and review the role of this drug and other therapeutic modalities in the management of vascular lesions. A seven month-old child with giant hemangioma and persistent bleeding, anemia and repeated infections was treated with interferon alpha-2a. The drug was administered at the dosage of 3 million units/m(2)/day, subcutaneously, for nine months. Response to treatment was considered excellent, with regression of 90% of the lesion, control of bleeding, infection and anemia, and catch-up growth. No untoward effects were observed. Interferon alpha-2a should be considered as a valid therapeutic option in selected cases of hemangioma in children.


Restricted Nitrogen and Water Applications in the Orchard Modify the Carbohydrate and Amino Acid Composition of Nonpareil and Carmel Almond Hulls.
Hull rot disease of almond (Prunus dulcis), caused by the fungus Rhizopus stolonifer, is prevalent in well maintained orchards where trees are provided plenty of water and nitrogen to increase the growth and yield. The predominantly grown variety Nonpareil is considered very susceptible to hull rot, while the pollinator variety Carmel is more resistant. Reduced nitrogen rates and restricted irrigation scheduling decreased the incidence and severity of hull rot in Californian orchards. As a part of our research, the hull composition of Australian almond fruits of Nonpareil and Carmel varieties, grown under two levels of irrigation (high and low) and two levels of nitrogen (high and low), were analysed using 1H NMR-based metabolomics. Both Nonpareil and Carmel hulls contained sugars such as glucose, sucrose, fructose and xylose, and amino acids, particularly asparagine. Variety was the major factor with Nonpareil hulls significantly higher in sugars and asparagine than Carmel. Within varieties, nitrogen influenced the relative concentrations of glucose, sucrose and asparagine. In Nonpareil, high nitrogen high water (the control) had relatively high glucose and asparagine content. High nitrogen low water increased the sucrose component, low nitrogen high water increased the glucose component and low nitrogen low water increased the sucrose and asparagine components. In Carmel, however, high nitrogen low water and low nitrogen high water increased sucrose and asparagine, and low nitrogen low water increased sucrose and glucose. Hull rot symptoms are caused by fumaric acid production by R. stolonifer growing within the hull. These changes in the hull composition under different nitrogen and water scenarios have the potential to affect the growth of R. stolonifer and its metabolite production in hull rot disease.


Resveratrol attenuates denervation-induced muscle atrophy due to the blockade of atrogin-1 and p62 accumulation.
Decrease in activity stress induces skeletal muscle atrophy. A previous study showed that treatment with resveratrol inhibits muscular atrophy in mdx mice, a model of DMD. However, almost all studies using resveratrol supplementation have only looked at adaptive changes in the muscle weight. The present study was designed to elucidate whether the resveratrol-inducing attenuation of skeletal muscle actually reflects the adaptation of muscle fibers themselves, based on the modulation of atrogin-1- or p62-dependent signaling. Mice were fed either a normal diet or 0.5% resveratrol diet. One week later, the right sciatic nerve was cut. The wet weight, mean fiber area, and amount of atrogin-1 and p62 proteins were examined in the gastrocnemius muscle at 14 days after denervation. The 0.5% resveratrol diet significantly prevented denervation-induced decreases in both the muscle weight and fiber atrophy. In addition, dietary resveratrol suppressed the denervation-induced atrogin-1 and p62 immunoreactivity. In contrast, 0.5% resveratrol supplementation did not significantly modulate the total protein amount of atrogin-1 or p62 in the denervated muscle of mice. Resveratrol supplementation significantly prevents muscle atrophy after denervation in mice, possibly due to the decrease in atrogin-1 and p62-dependent signaling.


High content screening for compounds that induce early stages of human embryonic stem cell differentiation.
Embryonic stem cells, due to their self-renewal and pluripotency properties, can be used to repair damaged tissues and as an unlimited source of differentiated cells. Although stem cells represent an important opportunity for cell based therapy and small molecules screening (in the context of drug or target discovery) many drawbacks are still preventing their widespread use. One of the most significant limitations is related to the complexity, as well as the reliability, of current protocols driving stem cells into any homogeneously differentiated cellular population. In this respect there is a strong demand for molecular agents promoting differentiation and thereby enabling robust, efficient and safe production of differentiated cells. In order to identify novel molecules that enhance early stages of differentiation, we developed an image based high content screening (HCS) approach using human embryonic stem cells (hESC). In our approach, we took advantage of custom image mining software specifically adapted for the selection of stem cell differentiation agents and the rejection of false positive hits. As a proof of concept -3500 small molecules originating from commercial libraries were screened and a number of molecules of interests were identified. These molecules show stem cell differentiation properties comparable to the phenotypic signature obtained with the reference compound retinoic acid.


Circulatory reaction to heat and cold in vibration-induced white finger with and without sympathetic blockade--an experimental study.
Central nervous, sympathetic mechanisms and local factors influencing the vascular response to cold in the finger skin were studied with plethysmographic and laser-Doppler methods in a chamber with controlled temperatures from 45 to about 5 degrees C, with and without sympathetic finger blockade. Blood flow and pressure were assessed after finger occlusion, together with finger heating and cooling, in workers with vibration-induced white finger (VWF) and in healthy persons not exposed to cold or vibration. Finger blood flow at vasodilatation, as measured by finger arterial inflow (FAI) after occlusion was smaller in the VWF group than in the reference subjects both with and without anesthesia, and capillary flux in the anesthetized finger tended to be smaller. At vasoconstriction, with or without anesthesia, the VWF subjects had lower FAI values than the referents. The studied flow and pressure variables differed considerably between the groups. The peripheral resistance of the VWF subjects was higher than that of the referents. This difference was the most pronounced after finger anesthesia. A possible pathogenetic mechanism in VWF is an increase in peripheral resistance due to a local defect in the vessels, with subsequent reduction in flow and intramural pressure.


Strategies used in the detection of acute/early HIV infections. The NIMH Multisite Acute HIV Infection Study: I.
Acute/early HIV infection plays a critical role in onward HIV transmission. Detection of HIV infections during this period provides an important early opportunity to offer interventions which may prevent further transmission. In six US cities, persons with acute/early HIV infection were identified using either HIV RNA testing of pooled sera from persons screened HIV antibody negative or through clinical referral of persons with acute or early infections. Fifty-one cases were identified and 34 (68%) were enrolled into the study; 28 (82%) were acute infections and 6 (18%) were early infections. Of those enrolled, 13 (38%) were identified through HIV pooled testing of 7,633 HIV antibody negative sera and 21 (62%) through referral. Both strategies identified cases that would have been missed under current HIV testing and counseling protocols. Efforts to identify newly infected persons should target specific populations and geographic areas based on knowledge of the local epidemiology of incident infections.


Effects of some steroids and other compounds on ovarian growth of the red swamp crayfish, Procambarus clarkii, during early vitellogenesis.
The effects of spiperone, a dopaminergic receptor blocker, the terpenoid JHIII, and the steroids 17alpha-hydroxyprogesterone and 17beta-estradiol (10(-7) mol/crayfish) were assayed in vivo on females of the red swamp crayfish, Procambarus clarkii, during early vitellogenesis. All these compounds were injected twice a week for 3 weeks. The rate of molting in response to these compounds was also determined. At the end of the experiment the ovaries were dissected out, in order to determine the gonadosomatic indexes and oocyte diameters. Spiperone, 17alpha-hydroxyprogesterone, and 17beta-estradiol produced a significant increase in the gonadosomatic index, while only the first two compounds were able to significantly increase the oocyte diameter; therefore, the effect of 17beta-estradiol the on gonadosomatic index could be merely due to increased water uptake by the ovary. Spiperone could be causing a decrease in secretion of the gonad and/or mandibular inhibiting hormones from the eyestalks, and 17alpha-hydroxyprogesterone could be having a direct stimulatory effect on vitellogenin production. Concerning molting, of the compound tested, only JHIII had a significant (P < 0.05) stimulatory effect, producing a 100% molting rate versus 66.7% among the controls.


Effectiveness of a presentation on infant oral health care for parents.
The aim of this study was to evaluate an infant oral health education programme, using a pre-post test design, for parents attending a paediatric clinic. The subjects were parents attending the well baby appointments at 3, 6, and 9 months of age. The study participants were men and women, all with an infant between 3 and 12 months of age. A 16 question assessment in the form of a questionnaire was completed immediately before and after the introduction of a 30 min educational intervention in the form of a PowerPoint presentation and a video of infant oral hygiene for parents. The parents completed the questionnaire twice (pre-post test design) in the same visit. Recruited parents attended only one presentation. The presentation educated parents about infant oral health and provided anticipatory guidance. Forty-seven parents or caretakers participated in the study. On the pre-test 28% had a score of 70% or less, and on the post-test 87% got a score of 88% or better. On the pre-test, 72% had a score of 70% or higher, and on the post-test 87% got a score of 88% or higher. Most parents (80%) reported that the presentation was helpful and indicated that the information would change the way they care for their baby's teeth at home. This study demonstrated the effectiveness of a 30 min PowerPoint and Video presentation in improving the oral health knowledge of parents caring for an infant.


Dynamics of RNA modification by a multi-site-specific tRNA methyltransferase.
In most organisms, the widely conserved 1-methyl-adenosine58 (m1A58) tRNA modification is catalyzed by an S-adenosyl-L-methionine (SAM)-dependent, site-specific enzyme TrmI. In archaea, TrmI also methylates the adjacent adenine 57, m1A57 being an obligatory intermediate of 1-methyl-inosine57 formation. To study this multi-site specificity, we used three oligoribonucleotide substrates of Pyrococcus abyssi TrmI (PabTrmI) containing a fluorescent 2-aminopurine (2-AP) at the two target positions and followed the RNA binding kinetics and methylation reactions by stopped-flow and mass spectrometry. PabTrmI did not modify 2-AP but methylated the adjacent target adenine. 2-AP seriously impaired the methylation of A57 but not A58, confirming that PabTrmI methylates efficiently the first adenine of the A57A58A59 sequence. PabTrmI binding provoked a rapid increase of fluorescence, attributed to base unstacking in the environment of 2-AP. Then, a slow decrease was observed only with 2-AP at position 57 and SAM, suggesting that m1A58 formation triggers RNA release. A model of the protein-tRNA complex shows both target adenines in proximity of SAM and emphasizes no major tRNA conformational change except base flipping during the reaction. The solvent accessibility of the SAM pocket is not affected by the tRNA, thereby enabling S-adenosyl-L-homocysteine to be replaced by SAM without prior release of monomethylated tRNA.


Familial retinoblastoma and chromosome 13 deletion transmitted via an insertional translocation.
Surviving persons from a kindred in which retinoblastoma occurred over four generations, transmitted by eight unaffected individuals, underwent chromosomal analysis. The results revealed that the development of retinoblastoma was associated with a constitutional chromosome deletion del(13)(q13.1q14.5) and that the unaffected transmitting state was associated with a balanced insertional translocation. These findings indicate that predisposition to retinoblastoma may be attributed to the loss of specific genetic material and that a chromosomal mechanism may explain apparent lack of gene penetrance in certain families. The development of unilateral, and not bilateral, retinoblastoma suggests either that the chromosome deletion is different from the mutation of heritable retinoblastoma in general, or that the chromosome deletion lessens the probability of subsequent somatic carcinogenic events.


Teacher Coaching: Increasing Deaf Students' Active Engagement Through Flexible Instructional Arrangements.
The study examined the effects of a coaching intervention on teachers' ability to implement academically responsive instruction through flexible instructional arrangements in self-contained classrooms for students who are deaf and hard of hearing, as well as the impact of instructional arrangements on students' academic engagement. Using a changing criterion design replicated across teachers, three-teacher participants with diverse backgrounds received differentiated coaching to implement flexible instructional arrangements. Results showed that coaching had an impact on all three teachers' implementation of flexible instructional arrangements. Concomitantly, students increased their active engagement and decreased passive engagement when they spent less time in whole class and more time in small group and child-managed arrangements. Teachers maintained the use of flexible instructional arrangements and students continued to be more actively engaged than pre-intervention. Limitations and implications for practice and research are discussed.


Laboratory Findings in Patients with Probable Dengue Diagnosis from an Endemic Area in Colombia in 2018.
As demonstrated with the novel coronavirus pandemic, rapid and accurate diagnosis is key to determine the clinical characteristic of a disease and to improve vaccine development. Once the infected person is identified, hematological findings may be used to predict disease outcome and offer the correct treatment. Rapid and accurate diagnosis and clinical parameters are pivotal to track infections during clinical trials and set protection status. This is also applicable for re-emerging diseases like dengue fever, which causes outbreaks in Asia and Latin America every 4 to 5 years. Some areas in the US are also endemic for the transmission of dengue virus (DENV), the causal agent of dengue fever. However, significant number of DENV infections in rural areas are diagnosed solely by clinical and hematological findings because of the lack of availability of ELISA or PCR-based tests or the infrastructure to implement them in the near future. Rapid diagnostic tests (RDT) are a less sensitive, yet they represent a timely way of detecting DENV infections. The purpose of this study was to determine whether there is an association between hematological findings and the probability for an NS1-based DENV RDT to detect the DENV NS1 antigen. We also aimed to describe the hematological parameters that are associated with the diagnosis through each test.


Effects of mesenchymal stem cells on Treg cells in rats with colitis.
The aim was to investigate the therapeutic effect of bone marrow mesenchymal stem cells (BM-MSC) on dextran sulfate sodium (DSS) induced colitis in rats and its effect on regulatory T cells (Treg). A model of DSS-induced colitis was established. BM-MSC was isolated and cultured to observe the efficacy of BM-MSC on colitis, including general vital signs, weight changes, colonic length changes, colonic histopathological changes, and colonic tissue MPO activity. The expression of inflammatory factors (IFN-γ, IL-4, IL-17, TGF-β) in colonic tissues was measured by real-time PCR. The amount of CD4 + CD25 + Treg was detected by flow cytometry. Real-time PCR was used to detect Foxp3+mRNA in CD4 + CD25 + Treg, western to detect Foxp3+protein expression in CD4 + CD25 + Treg, and ELISA was used to detect IL-35 and IL-10 cytokines in CD4 + CD25 + Treg culture supernatant. Results show that intravenous injection of BM-MSC significantly improved the clinical manifestations and histopathological changes in rats with experimental DSS colitis; significantly down-regulated the expression of inflammatory factors IFN-γ, IL-4, and IL-17 and up-regulated the expression of TGF-β in colon tissues; BM-MSC also increased the number of CD4+CD25+Foxp3+Treg and enhanced the function of CD4+CD25+Foxp3+Treg in colon tissues, and up-regulated the expression of IL-35. In conclusion, BM-MSC has a certain therapeutic effect on DSS-induced colitis. It can improve the general signs of colitis rats and reduce intestinal injury and inflammatory response. The immunoregulatory effect of BM-MSC is achieved by enhancing the function of CD4+CD25+Foxp3+Treg and up-regulating the secretion of immunosuppressive inflammatory factors.


Analysis of the Effect Size of Overweight in Muscular Strength Tests Among Adolescents: Reference Values According to Sex, Age, and Body Mass Index.
Martínez-López, EJ, De La Torre-Cruz, M, Suárez-Manzano, S, and Ruiz-Ariza, A. Analysis of the effect size of overweight in muscular strength tests among adolescents: reference values according to sex, age, and body mass index. J Strength Cond Res 32(5): 1404-1414, 2018-The aim of this paper is to quantify the effect size of overweight on the results of muscular strength tests in adolescents and to report percentile tables based on sex, age, and body mass index (BMI). The study hypothesized that the effect size obtained from the difference between normal-weight and overweight youth would be greater than the differences between sexes within the same age group. A total of 11,044 Spanish adolescents (48.5% girls) aged 14.39 ± 1.21 years (range: 12-16 years) from 42 secondary schools participated in the research. Muscular strength was evaluated using standing long jump, hand grip strength (manual dynamometer), and sit-ups (30 seconds). The effect size was analyzed using the adjusted Hedges' ğ. The results show that 76.3 and 72.8% of overweight boys and girls, respectively, performed a standing long jump equal to or less than the normal-weight average. The 67.4 and 67.1% of overweight boys and girls, respectively, showed manual dynamometer values equal to or greater than the normal-weight average. Finally, 68.7 and 65.9% of overweight boys and girls, respectively, obtained measures for 30 seconds of sit-ups equal to or lower than the normal-weight average. It can therefore be concluded that the differential effect size between boys and girls is higher than that between normal-weight and overweight adolescents in the 3 strength tests analyzed. Despite the above, these results suggest the value of taking into account the BMI when assessing the muscular strength of young people, in addition to sex and age.


Nuisance parameter elimination for proportional likelihood ratio models with nonignorable missingness and random truncation.
We show that the proportional likelihood ratio model proposed recently by Luo & Tsai (2012) enjoys model-invariant properties under certain forms of nonignorable missing mechanisms and randomly double-truncated data, so that target parameters in the population can be estimated consistently from those biased samples. We also construct an alternative estimator for the target parameters by maximizing a pseudo-likelihood that eliminates a functional nuisance parameter in the model. The corresponding estimating equation has a U-statistic structure. As an added advantage of the proposed method, a simple score-type test is developed to test a null hypothesis on the regression coefficients. Simulations show that the proposed estimator has a small-sample efficiency similar to that of the nonparametric likelihood estimator and performs well for certain nonignorable missing data problems.


Neutralizing TNF-alpha activity modulates T-cell phenotype and function in experimental autoimmune uveoretinitis.
Inhibiting TNF-alpha activity prevents tissue destruction without inhibiting retinal T cell infiltration in experimental autoimmune uveoretinitis (EAU) in Lewis rats. To further determine the role of TNF-alpha in autoimmune uveitis we characterized T cells isolated from retinae after treatment with a TNF-alpha antagonist. TNF-alpha activity was neutralized in vivo with a p55 TNF-alpha receptor-Ig fusion protein (sTNFr-Ig), administered 8 and 10 days after induction of EAU with heterologous retinal antigens. Retinal T-cell phenotype expression was examined by flow cytometry with respect to OX22 status (CD45RBlow or CD45RBhigh), activation (OX40 and CD25 expression) and rate of T-cell apoptosis (Annexin V+PI- expression). Lymphocyte reactivity was assessed by proliferation responses and cytokine production to retinal antigens. Despite greater than 40% of CD4+ T cells being activated at the height of disease, the proportion of OX22low expression was reduced and T cells exhibited reduced IFN-gamma and elevated IL-4 production. Retinal T cells maintained antigen-specific proliferation and demonstrated a low apoptotic rate. Although in both animal groups, comparable numbers of T cells were isolated, neutralizing TNF activity suppressed Th1 effector mechanisms protecting against target organ damage.


Universal TA cloning.
TA cloning is one of the simplest and most efficient methods for the cloning of PCR products. The procedure exploits the terminal transferase activity of certain thermophilic DNA polymerases, including Thermus aquaticus (Taq) polymerase. Taq polymerase has non-template dependent activity which preferentially adds a single adenosine to the 3'-ends of a double stranded DNA molecule, and thus most of the molecules PCR amplified by Taq polymerase possess single 3'-A overhangs. The use of a linearized "T-vector" which has single 3'-T overhangs on both ends allows direct, high-efficiency cloning of PCR products, facilitated by complementarity between the PCR product 3'-A overhangs and vector 3'-T overhangs. The TA cloning method can be easily modified so that the same T-vector can be used to clone any double-stranded DNA fragment, including PCR products amplified by any DNA polymerase, as well as all blunt- and sticky-ended DNA species. This technique is especially useful when compatible restriction sites are not available for the subcloning of DNA fragments from one vector to another. Directional cloning is made possible by appropriate hemi-phosphorylation of both the T-vectors and the inserts. With a single T-vector at hand, any DNA fragment can be cloned without compromising the cloning efficiency. The universal TA cloning method is thus both convenient and labor-saving.


MOVEMENT IN THE CYANOPHYCEAE : THE EFFECT OF pH UPON MOVEMENT IN OSCILLATORIA.
The effect of pH upon the velocity of translatory movement of Oscillatoria formosa Bory in inorganic culture solutions was determined. Unhindered movement occurred in the range of about pH 6.4 to 9.5. Above and below these limits inhibition was marked. In the unfavorable acid and alkaline ranges inhibition was progressive with exposure time; in the favorable range continuous movement was maintained for 24 hours.


Coexpression and heteromerization of two neuronal K-Cl cotransporter isoforms in neonatal brain.
The neuron-specific K-Cl cotransporter KCC2 maintains the low intracellular chloride concentration required for the fast hyperpolarizing actions of inhibitory neurotransmitters. The KCC2 gene codes for two isoforms, KCC2a and KCC2b, which differ in their N termini. The relative expression and cellular distribution of the two KCC2 protein isoforms are unknown. Here, we characterize an antibody against the KCC2a isoform and show that a previously described antibody against KCC2 is specific for the KCC2b isoform (Hubner, C. A., Stein, V., Hermans-Borgmeyer, I., Meyer, T., Ballanyi, K., and Jentsch, T. J. (2001) Neuron 30, 515-524). Immunostaining of dissociated hippocampal cultures confirms that both KCC2 isoforms are neuron-specific. Immunoblot analysis indicates that KCC2b is the major KCC2 isoform in the adult brain, whereas in the neonatal mouse central nervous system, half of total KCC2 protein is KCC2a. At this stage, the two KCC2 isoforms are largely colocalized and show similar patterns of distribution in the brain. When coexpressed in HEK293 cells, KCC2a and KCC2b proteins form heteromeric complexes. Moreover, the two isoforms can be coimmunoprecipitated from the neonatal brain, suggesting the presence of endogenous KCC2a-KCC2b heteromers. Consistent with this, native gel analysis shows that a substantial part of endogenous KCC2 isoforms in the neonatal brain constitute dimers.