The organization and distribution of patient education materials in family medicine practices.
Millions of dollars are spent annually on the production and distribution of patient education materials; however, there are no studies describing their actual use by physicians. Using qualitative data from a large comparative case study, our analysis evaluates how patient education materials are organized and used in family practices. Eighteen purposefully selected family medicine practices were directly observed for 4 to 12 weeks each. A total of 57 providers were shadowed by a research nurse, and detailed field notes on 1600 patient encounters were recorded. A 3-member analysis team reviewed the qualitative data and identified emergent patterns. Clinics' use of patient education materials fell mostly into 2 distinct patterns. "Stockpilers" were providers who relied on the clinic staff to develop and organize a common library of patient education handouts. Providers with a "personal stash" collected much smaller numbers of materials that they personally maintained. Providers in the latter group had a known repertoire of a limited amount of educational material and used it more often than providers with access to a greater variety and number of handouts. In all practices, providers distributed most handouts; staff and self-selection by patients played a minor role. It appears that provider involvement and familiarity with patient education materials are key to their use in clinical practice. Clinicians use written patient education materials most efficiently by personally selecting and maintaining a small number of handouts that address topics most relevant to their practice.


Identification of structural determinants of the first transmembrane domain of the small envelope protein of duck hepatitis B virus essential for particle morphogenesis.
The envelope of duck hepatitis B virus (DHBV) consists of the small (S) and large (L) envelope proteins, which share a common C-terminal multispanning transmembrane region but differ by the long N-terminal pre-S domain of L, which is essential for interactions with both the receptor and nucleocapsid. To achieve these dual functions, L acquires mixed topologies through S-dependent post-translational translocation of its pre-S domain. This study has examined the role of S in this unusual mechanism of translocation by analysis of the alpha-helical transmembrane domains and their potential to engage in lateral interactions for envelope assembly. Through mutagenesis in constructs expressing the S and L envelope proteins independently, transmembrane domain 1 was identified as an essential structural determinant in S. Two polar residues in this helix were identified as contributing to L protein translocation through the assembly of S into particles, implying that the topological switch of L is part of the assembly and maturation process. The same domain in L was shown to be dispensable for L translocation and assembly, suggesting that transmembrane domain 1 of L and S have different functional roles and structural arrangements on the assembled particle. The conservation in all hepadnavirus envelope proteins of two polar residues at positions 24 and 27 of transmembrane domain 1, the former positively charged, points to this being a common determinant in particle morphogenesis for all hepadnaviruses.


Hidden Toxicity in Neonatal Intensive Care Units: Phthalate Exposure in Very Low Birth Weight Infants.
To determine exposure to endocrine-disrupting phthalates in preterm infants in neonatal intensive care units (NICU). Urine samples (n=151) from 36 preterm infants (<32 weeks of gestation and/or <1500 g of birth weight) were collected on the first 3 days of admission to the NICU and biweekly thereafter. Diethylhexyl phthalate contents of indwelling medical devices used in various procedures and the concentrations of phthalate metabolites in the urine samples were analyzed. The relationships between urinary excretion, exposure intensity, postnatal age and birth weight were examined. The mean gestational age and mean birth weight of the study infants were 28.9±1.5 weeks and 1024±262 g, respectively. Diethylhexyl phthalate was detected in umbilical catheters, endotracheal tubes, nasogastric tubes, and nasal cannula. Monoethylhydroxyhexyl phthalate (MEHHP) was the most frequently detected metabolite (81.4%); its concentration increased during the first 4 weeks and then started to decrease but never disappeared. Patients who did not need indwelling catheters (except nasogastric tubes) after 2 weeks were classified as group 1 and those who continued to have indwelling catheters as group 2. Although not of statistical significance, MEHHP levels decreased in group 1 but continued to stay high in group 2 (in the 4th week, group 1: 65.9 ng/mL and group 2: 255.3 ng/mL). Levels of MEHHP in the first urinary samples were significantly higher in infants with a birth weight <1000 g (<1000 g: 63.2±93.8 ng/mL, ≥1000 g: 10.9±22.9 ng/mL, p=0.001). Phthalate metabolites were detected even in the first urine samples of very low birth weight newborns. Phthalate levels were higher in the first weeks of intensive invasive procedures and in preterm infants with a birth weight less than 1000 g. MEHHP was the most frequently detected metabolite and could be a suitable biomarker for the detection of phthalate exposure in preterm infants.


A New Entrance Technique for C2 Pedicle Screw Placement and the Use in Patients With Atlantoaxial Instability.
A prospective study and a technique note. To introduce a new entrance technique for C2 pedicle screw placement and to measure the related linear and angular parameters about the entrance point on computed tomography (CT) images. The safety of this technique for patients with atlantoaxial instability was also evaluated. Although earlier studies have introduced different methods for C2 pedicle screw placement, the entry points and the angular parameters may be variable. Few studies have established a fixed entry point on the basis of the anatomic structure of C2 for pedicle screw placement. A total of 60 dry C2 vertebrae were obtained for anatomic measurement in the study. The posterior bilateral nutrient foramens of C2 lamina were selected as the entry points for pedicle screw placement. The foramens were marked with needles and then the vertebrae underwent CT scan. The axial and sagittal planes of C2 pedicles were harvested and 4 linear and 2 angular parameters about the entry point were determined. After that, we used the entrance technique on 31 patients with atlantoaxial instability in a prospective study. CT of the cervical spine was performed to evaluate the safety of the entrance technique. The nutrient foramens exist in 97% of the left lamina and 93% of the right lamina of the C2 vertebra. The overall mean distance from the entry point (nutrient foramen) to the superior border of lamina (PSD), to the inferior border of lamina (PID), to the medial border of the pedicle (PMD), and the length of pedicle screw trajectory (PL, transit the pedicle center) were 3.32±0.63, 8.33±1.21, 6.85±1.00, and 24.47±1.51, respectively. The averaged transverse angle (α) on the axial plane and the superior angle (β) on the sagittal plane were 19.83±3.83 and 30.12±6.02 degrees, respectively. Then, 31 patients underwent bilateral C2 pedicle screw fixation without screw violation into the spinal canal or vertebral artery injury by the new entrance technique. The overall mean angles α and β and the length of the pedicle screw were 17.52±3.81 and 34.29±4.18 degrees and 25.85±2.06 mm, respectively. No statistical differences were found in these 3 parameters between the dry C2 vertebrae and the C2 vertebrae of patients who underwent the surgery (P>0.05). Using the posterior bilateral nutrient foramens of the C2 lamina as the entry point is a helpful intraoperative landmark for C2 pedicle screw placement.


Breeding of a non-obese, diabetic strain of mice.
A female mouse spontaneously exhibiting polyuria and glucosuria accompanied by rapid weight loss waa found in one of two sublines derived from the CTS mice. Eight mating pairs were made using its offspring and selection was performed for both spontaneous diabetes and reproductive ability. After six generations of the selective breeding the diabetic (nod) and the control (non) lines were established. A marked sex difference was observed in the incidence of diabetic symptoms in the nod mouse. The cumulative incidence of the onset up to 30 weeks of age was 80% in females and less than 20% in males. The onset of diabetes was abrupt in both sexes, and spontaneous remission was not observed. However, daily administration of insulin induced an increase of body weight and a prolongation of life span. Diabetic symptoms are biochemically characterized by polyuria, polydipsia, hyperglycemia, glucosuria and hypercholesteremia. Pathological examination revealed a high frequency of lymphocyte infiltration around and/or into the Langerhans' islet. It was observed even at the prediabetic stage over five weeks of both sexes. The number and size of the islets were markedly reduced in the overt diabetic mice. Although the mechanism of the pathogenesis is not clear yet, the nod mouse may be a useful animal model for investigating the human juvenile type diabetes.


Component removal in revision total hip arthroplasty.
Component removal is a time-consuming yet necessary step during revision hip surgeries. Because of the diversity of the components and the methods used to secure them, an equal diversity of approaches and tools are necessary for component removal. Careful and detailed preoperative planning is mandatory, the mode of failure must be understood, and detailed imaging should be available to the surgeon. Understanding the basic principles and indications for each of the techniques would optimize outcome. We review the approaches, tools, and techniques for component removal in revision hip procedures in stepwise sequence.


Disruption of Th2a and Th2b genes causes defects in spermatogenesis.
The variant histones TH2A and TH2B are abundant in the testis, but their roles in spermatogenesis remain elusive. Here, we show that male mutant mice lacking both Th2a and Th2b genes were sterile, with few sperm in the epididymis. In the mutant testis, the lack of TH2B was compensated for by overexpression of H2B, whereas overexpression of H2A was not observed, indicating a decrease in the total histone level. Mutant mice exhibited two defects: incomplete release of cohesin at interkinesis after meiosis I and histone replacement during spermiogenesis. In the mutant testis, secondary spermatocytes at interkinesis accumulated and cohesin was not released normally, suggesting that the retained cohesion of sister chromatids delayed the subsequent entry into meiosis II. In addition, impaired chromatin incorporation of TNP2 and degenerated spermatids were observed in the mutant testis. These results suggest that a loss of TH2A and TH2B function in chromatin dynamics or a decrease in the total histone levels causes defects in both cohesin release and histone replacement during spermatogenesis.


Loss of striatal mu1 opiate binding by substantia nigra lesions in the rat.
Opiate receptors have been identified within the striatum and some have been localized presynaptically to nigrostriatal neurons. Using unilateral ablative lesions of the substantia nigra, we examined binding in the ipsilateral and contralateral striata. Lesions significantly lowered both 3H[D-Ala2,MePhe4,Gly(ol)5]enkephalin (DAGO) and 3H[D-Ala2,Leu5]enkephalin (DADL) binding. The inclusion of competitors in these assays revealed a decrease in both mu1 and mu2 receptors. Mu1 binding was slightly more sensitive to the lesioning than mu2 binding. Selective mu1 and mu2 binding assays supported these observations. No change in delta binding was observed in the lesioned striata. These studies raise the possibility that both mu1 and mu2, but not delta, receptors are localized presynaptically on nigrostriatal neurons.


Comparison of seven tests for high-grade cervical intraepithelial neoplasia in women with abnormal smears: the Predictors 2 study.
High-risk human papillomavirus (HPV) DNA/RNA testing provides higher sensitivity but lower specificity than cytology for the identification of high-grade cervical intraepithelial neoplasia (CIN). Several new HPV tests are now available for this purpose, and a direct comparison of their properties is needed. Seven tests were evaluated with samples in liquid PreservCyt transport medium from 1,099 women referred for colposcopy: the Hybrid Capture 2 (Qiagen), Cobas (Roche), PreTect HPV-Proofer (NorChip), Aptima HPV (Gen-Probe), and Abbott RealTime assays, the BD HPV test, and CINtec p16(INK4a) cytology (mtm laboratories) immunocytochemistry. Sensitivity, specificity, and positive predictive value (PPV) were based on the worst histology found on either the biopsy or the treatment specimen after central review. Three hundred fifty-nine women (32.7%) had CIN grade 2+ (CIN2+), with 224 (20.4%) having CIN3+. For detection of CIN2+, Hybrid Capture 2 had 96.3% sensitivity, 19.5% specificity, and 37.4% PPV. Cobas had 95.2% sensitivity, 24.0% specificity, and 37.6% PPV. The BD HPV test had 95.0% sensitivity, 24.2% specificity, and 37.8% PPV. Abbott RealTime had 93.3% sensitivity, 27.3% specificity, and 38.2% PPV. Aptima had 95.3% sensitivity, 28.8% specificity, and 39.3% PPV. PreTect HPV-Proofer had 74.1% sensitivity, 70.8% specificity, and 55.4% PPV. CINtec p16(INK4a) cytology had 85.7% sensitivity, 54.7% specificity, and 49.1% PPV. Cytology of a specimen taken at colposcopy (mild dyskaryosis or worse) had 88.9% sensitivity, 58.1% specificity, and 50.7% PPV. Our study confirms that, in a referral setting, HPV testing by a number of different tests provides high sensitivity for high-grade disease. Further work is needed to confirm these findings in a routine screening setting.


Variation in small bowel biopsy performance among diverse endoscopy settings: results from a national endoscopic database.
The diagnostic gold standard for celiac sprue requires performance of small bowel biopsy to identify characteristic mucosal changes. The current biopsy practice among endoscopists for celiac sprue is unknown. The CORI (Clinical Outcomes Research Initiative) national endoscopic database was analyzed to determine the rate of small bowel biopsy in patients undergoing EGD for the evaluation of anemia, iron deficiency without anemia, weight loss, and diarrhea. Between January 2000 and September 2003, 3,992 patients underwent EGD for the evaluation of anemia, iron deficiency, weight loss, and diarrhea with normal EGD findings. Overall, 438 patients (11%) underwent small bowel biopsy. Small bowel biopsy performance varied with procedure indication: anemia 10%, iron deficiency 7%, weight loss 6%, and diarrhea 19%, p < 0.0001. Biopsy rates declined with advancing patient age; 12% (<50 yr), 12% (50-69 yr), and 9% (> or =70 yr), p= 0.002, and also differed with patient race (12%, Whites vs 8%, non-Whites), p= 0.0007. Biopsy performance varied among endoscopy sites: community (9%), academic (13%), and VAMC (15%), p < 0.0001. Multivariate analysis identified age > or =70 yr (OR: 0.52, p= 0.0007), non-White race (OR: 0.60, p= 0.0007) and an indication of weight loss (OR: 0.60, p= 0.01) to be associated with not performing a biopsy. Among diverse endoscopy sites, the majority of patients with anemia, iron deficiency, weight loss, and diarrhea do not undergo small bowel biopsy at EGD. The diagnosis of celiac sprue should be considered in such cases. Endoscopy provides an opportunity to obtain tissue to diagnose sprue, and should be strongly considered in this setting.


Cannabinoid receptor type 1 protects nigrostriatal dopaminergic neurons against MPTP neurotoxicity by inhibiting microglial activation.
This study examined whether the cannabinoid receptor type 1 (CB(1)) receptor contributes to the survival of nigrostriatal dopaminergic (DA) neurons in the 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) mouse model of Parkinson's disease. MPTP induced significant loss of nigrostriatal DA neurons and microglial activation in the substantia nigra (SN), visualized with tyrosine hydroxylase or macrophage Ag complex-1 immunohistochemistry. Real-time PCR, ELISA, Western blotting, and immunohistochemistry disclosed upregulation of proinflammatory cytokines, activation of microglial NADPH oxidase, and subsequent reactive oxygen species production and oxidative damage of DNA and proteins in MPTP-treated SN, resulting in degeneration of DA neurons. Conversely, treatment with nonselective cannabinoid receptor agonists (WIN55,212-2 and HU210) led to increased survival of DA neurons in the SN, their fibers and dopamine levels in the striatum, and improved motor function. This neuroprotection by cannabinoids was accompanied by suppression of NADPH oxidase reactive oxygen species production and reduced expression of proinflammatory cytokines from activated microglia. Interestingly, cannabinoids protected DA neurons against 1-methyl-4-phenyl-pyridinium neurotoxicity in cocultures of mesencephalic neurons and microglia, but not in neuron-enriched mesencephalic cultures devoid of microglia. The observed neuroprotection and inhibition of microglial activation were reversed upon treatment with CB(1) receptor selective antagonists AM251 and/or SR14,716A, confirming the involvement of the CB(1) receptor. The present in vivo and in vitro findings clearly indicate that the CB(1) receptor possesses anti-inflammatory properties and inhibits microglia-mediated oxidative stress. Our results collectively suggest that the cannabinoid system is beneficial for the treatment of Parkinson's disease and other disorders associated with neuroinflammation and microglia-derived oxidative damage.


Immune Characteristics of Patients with Coronavirus Disease 2019 (COVID-19).
Up to now, little is known about the detailed immune profiles of COVID-19 patients from admission to discharge. In this study we retrospectively reviewed the clinical and laboratory characteristics of 18 COVID-19 patients from January 30, 2020 to February 21, 2020. These patients were divided into two groups; group 1 had a severe acute respiratory syndrome coronavirus 2 nucleic acid-positive duration for more than 15 days (n = 6) and group 2 had a nucleic acid-positive duration for less than 15 days (n = 12). Group 1 patients had lower level of peripheral blood lymphocytes (0.40 vs. 0.78 ×109/L, p = 0.024) and serum potassium (3.36 vs. 3.79 mmol/L, p = 0.043) on admission but longer hospitalization days (23.17 vs. 15.75 days, p < 0.001) compared to Group 2 patients. Moreover, baseline level of lymphocytes (r = -0.62, p = 0.006) was negatively correlated with the nucleic acid-positive duration. Additionally, lymphocytes (420.83 vs. 1100.56 /μL), T cells (232.50 vs. 706.78 /μL), CD4+ T cells (114.67 vs. 410.44 /μL), and CD8+ T cells (94.83 vs. 257.44 /μL) in the peripheral blood analyzed by flow cytometry were significantly different between Group 1and Group 2. Furthermore, there was also a negative correlation between lymphocytes (r = -0.54, p = 0.038) or T cells (r = -0.55, p = 0.034) at diagnosis and the nucleic acid-positive duration, separately. In conclusion, the patients with nucleic acid-positive ≥ 15 days had significantly decreased lymphocytes, T cell and its subsets compared to those who remained positive for less than 15 days.


The Impact of Deterministic Signal Loss on OCT Angiography Measurements.
In this study, we aimed at investigating the impact of deterministic signal loss on image quality and, thus, on optical coherence tomography angiography (OCTA) measurements performed by the RTVue-XR Avanti System. Absorptive filters with different optical densities (ODs) were used to simulate signal loss in a controlled way in 30 eyes from 15 healthy subjects. Scan Quality (SQ), provided by the AngioVue software, was applied as a measure of image quality. Assessing the effect of decreased light transmittance on SQ values as well as that of attenuated image quality on OCTA parameters revealed a strong systematic dependence between these quantities. Attenuated image quality was associated with significantly decreased macular and peripapillary vessel density (VD) values, and we calculated a correction factor of 2.27% to 3.97% for a one-unit change in SQ for the different VD parameters. Our findings suggest that the influence of systematic changes in image quality on OCTA parameters needs to be considered during patient follow-up in order to make valid assessment of progression. For accurate evaluation of longitudinal changes in OCTA parameters, equal scan quality or using a correction factor is suggested.


In vitro activity of ceftaroline against clinical Staphylococcus aureus isolates collected during a national survey conducted in Belgian hospitals.
The aim of this study was to estimate the in vitro activity of ceftaroline against clinical Staphylococcus aureus isolates collected during national surveillance in Belgian acute-care hospitals. Ceftaroline-resistant isolates were further investigated for their resistance mechanisms. From October 2013 to March 2014, 155 laboratories of Belgian acute-care hospitals were invited to send to the National Reference Centre-Staphylococcus aureus (Belgium) up to five non-duplicate S. aureus including three MRSA and two MSSA from hospitalized patients. Isolates were analysed by spa typing, SCCmec typing (for MRSA) and PCR for detection of 16S-mecA-nuc and 16S-mecC. MICs of oxacillin, cefoxitin and ceftaroline were determined by the broth microdilution method. The nucleotide sequences of mecA, native pbp and gdpP genes of isolates with reduced susceptibility to ceftaroline were analysed for the presence of mutations responsible for amino acid substitutions. Ninety-nine percent of isolates, including MRSA (n = 284) and MSSA (n = 131), were susceptible to ceftaroline. Only four MRSA isolates showed resistance to ceftaroline (MIC = 2 mg/L). These four isolates belonged to lineages CC5 (n = 1), CC22 (n = 2) and CC8 (n = 1). Two isolates (CC22 and CC8) carried mutations in mecA, as well as in other pbp genes. The remaining isolates carried mutations in native pbp genes or in gdpP. This is the first Belgian in vitro survey on ceftaroline activity against S. aureus. This antibiotic showed excellent activity against MRSA and MSSA, and only a few MRSA isolates with resistance were found. Reduced susceptibility to ceftaroline seems a complex phenomenon due to the accumulation of mutations in genes involved in β-lactam tolerance.


Circadian rhythms in pump parameters of patients on contemporary left ventricular assist device support.
Algorithms to monitor pump parameters are needed to further improve outcomes after left ventricular assist device (LVAD) implantation. Previous research showed a restored circadian rhythm in pump parameters in patients on HeartWare (HVAD) support. Circadian patterns in HeartMate3 (HM3) were not studied before, but this is important for the development of LVAD monitoring algorithms. Hence, we aimed to describe circadian patterns in HM3 parameters and their relation to patterns in heart rate (HR). 18 HM3 patients were included in this study. HM3 data were retrieved at a high frequency (one sample per 1 or 2 h) for 1-2 weeks. HR was measured using a wearable biosensor. To study overall patterns in HM3 parameters and HR, a heatmap was created. A 24-h cosine was fitted on power and HR separately. The relationship between the amplitude of the fitted cosines of power and HR was calculated using Spearman correlation. A lower between patient variability was found in power compared with flow and PI. 83% of the patients showed a significant circadian rhythmicity in power (p < 0.001-0.04), with a clear morning increase. All patients showed significant circadian rhythmicity in HR (p < 0.001-0.02). The amplitudes of the circadian rhythm in power and HR were not correlated (Spearman correlation of 0.32, p = 0.19). A circadian rhythm of pump parameters is present in the majority of HM3 patients. Higher frequency pump parameter data should be collected, to enable early detection of complications in the future development of predictive algorithms.


Association between hyperinsulinemia and increased risk of cancer death in nonobese and obese people: A population-based observational study.
Obesity, metabolic syndrome and type 2 diabetes are associated with cancer-related mortality. We assessed whether hyperinsulinemia is a risk factor for cancer death in nonobese people without diabetes. We conducted a prospective cohort study using data from the National Health and Nutrition Examination Survey 1999-2010 and followed up the participants until December 31, 2011. For the primary analysis of cancer mortality, we used Cox proportional hazard models to estimate hazard ratios (HRs) in the participants with hyperinsulinemia and those without. Hyperinsulinemia was defined as a fasting insulin level of ≥10 μU/mL. To identify causes of deaths, the International Classification of Diseases, Tenth Revision codes were used. This study included 9,778 participants aged 20 years or older without diabetes or a history of cancer: 6,718 nonobese participants (2,057 with hyperinsulinemia [30.6%]) and 3,060 obese participants (2,303 with hyperinsulinemia [75.3%]). A total of 99.9% completed follow-up. Among all study participants, cancer mortality was significantly higher in those with hyperinsulinemia than in those without hyperinsulinemia (adjusted HR 2.04, 95% CI 1.24-3.34, p = 0.005). Similarly, among nonobese participants, multivariable analysis showed that cancer mortality was significantly higher in those with hyperinsulinemia than in those without (adjusted HR 1.89, 95% CI 1.07-3.35, p = 0.02). Considering that nonobese people with hyperinsulinemia were at higher risk of cancer mortality than those without hyperinsulinemia, improvement of hyperinsulinemia may be an important approach for preventing cancer regardless of the presence or absence of obesity.


Tobacco mosaic virus-directed reprogramming of auxin/indole acetic acid protein transcriptional responses enhances virus phloem loading.
Vascular phloem loading has long been recognized as an essential step in the establishment of a systemic virus infection. In this study, an interaction between the replication protein of tobacco mosaic virus (TMV) and phloem-specific auxin/indole acetic acid (Aux/IAA) transcriptional regulators was found to modulate virus phloem loading in an age-dependent manner. Promoter expression studies show that in mature tissues TMV 126/183-kDa-interacting Aux/IAAs predominantly express and accumulate within the nuclei of phloem companion cells (CCs). Furthermore, CC Aux/IAA nuclear localization is disrupted upon infection with an interacting virus. In situ analysis of virus spread shows that the inability to disrupt Aux/IAA CC nuclear localization correlates with a reduced ability to load into the vascular tissue. Subsequent systemic movement assays also demonstrate that a virus capable of disrupting Aux/IAA localization is significantly more competitive at moving out of older plant tissues than a noninteracting virus. Similarly, CC expression and overaccumulation of a degradation-resistant Aux/IAA-interacting protein was found to inhibit TMV accumulation and phloem loading selectively in flowering plants. Transcriptional expression studies demonstrate a role for Aux/IAA-interacting proteins in the regulation of salicylic and jasmonic acid host defense responses as well as virus-specific movement factors, including pectin methylesterase, that are involved in regulating plasmodesmata size-exclusion limits and promoting virus cell-to-cell movement. Combined, these findings indicate that TMV directs the reprogramming of auxin-regulated gene expression within the vascular phloem of mature tissues as a means to enhance phloem loading and systemic spread.


A pancreatic cancer mouse model with human immunity.
Pancreatic ductal adenocarcinoma (PDAC) is characterized by a tumor immune microenvironment (TIME) that promotes resistance to immunotherapy. A preclinical model system that facilitates studies of the TIME and its impact on the responsiveness of human PDAC to immunotherapies remains an unmet need. We report a novel mouse model, which develops metastatic human PDAC that becomes infiltrated by human immune cells recapitulating the TIME of human PDAC. The model may serve as a versatile platform to study the nature of human PDAC TIME and its response to various treatments.


Advancing age and the risk of nasal carriage of Staphylococcus aureus among patients on long-term hospital-based hemodialysis.
Elevated nasal carriage rates of Staphylococcus aureus and ensuing complications among the elderly and in those on long-term hemodialysis (HD) are well recognized. The aim of the present study was to determine the extent to which advancing age is associated with the risk of persistent S. aureus nasal carriage among end-stage renal disease (ESRD) patients on long-term HD. This prospective study involved 205 ESRD patients enrolled for maintenance HD from July 1997 to July 2000. Persistent nasal carriage was defined by two or more positive cultures for methicillin-susceptible (MSSA) or methicillin-resistant S. aureus (MRSA). Five standardized swabs were taken from the anterior nares of all the patients on long-term HD. S. aureus nasal carriage rates were estimated and compared among ESRD patients of different age groups. Overall, a prevalence of 38.05% (78/205) for S. aureus nasal carriage was observed, including 27.3% (56/205) for MSSA and 10.7% (22/205) for MRSA. Patients aged 75 to 84 years had the highest (84.6%, 11/13) prevalence of S. aureus nasal carriage (RR, 7.000, 95% CI, 4.350-11.763, P<0.00001). Those aged 65 to 74 years had the next highest (49.0%, 25/51) nasal carriage rates (RR, 4.083, 95% CI, 2.302-7.658, P<0.0001) while patients aged 15 to 24 years (reference group) had the lowest (12.8%, 1/8) prevalence of nasal carriage. The 75 to 84 year age group also had the highest rates of MSSA (46.2%, 6/13), (RR- 3.833, 95% CI, 2.144-7.234, P<0.0001) and MRSA (38.5 %, 5/13) (RR, 6.333, 95%CI, 2.767-16.198, P<0.0001) nasal carriage compared to the reference group. Significantly higher persistent MSSA and MRSA nasal carriage rates among ESRD patients >75 years of age are suggestive of an elevated risk of potentially serious S. aureus- related complications among the very elderly during long-term HD. These findings might be helpful in the identification of elderly HD patients as a high-risk group for S. aureus-linked vascular access-related septicemia (VRS) and to evolve appropriate preventive strategies.


Congenital diaphragmatic hernia beyond infancy.
Congenital diaphragmatic hernia (CDH) is a common cause of severe respiratory distress in the newborn. However, the presentation of CDH in older children and adults is rare, and, therefore, little is known concerning its symptoms, operative management, and postoperative complications. Thirteen patients (age range: 2 months to 26 years; 5 males, 8 females) presented with CDH. Four patients had right-sided hernias, eight left-sided hernias, and one bilateral hernias. Symptoms included chronic respiratory tract infections in 6 patients, vomiting in 5, weight loss in 1, severe failure to thrive in 2, and severe respiratory distress in 3; one patient was asymptomatic. Physical signs included the absence of breathing sounds or bowel sounds in the chest in eight patients, hyperresonance in one, and cachexia in two. The diagnosis was confirmed in each patient by chest roentgenogram or gastrointestinal contrast radiograph. All patients underwent immediate repair. After reduction of the viscera, 12 of 13 patients underwent primary diaphragm repair, whereas one patient required a prosthetic diaphragm patch. Twelve of 13 patients (92%) survived. Postoperatively, 7 of the 12 survivors (58%) developed severe gastric atony, and four required further operative therapy. In contrast to newborns, CDH in the older child and adult is frequently seen on the right side, rarely presents with severe respiratory distress, and is occasionally asymptomatic. Postoperative gastric atony is a major cause of morbidity, making transabdominal repair with simultaneous pyloroplasty and/or feeding jejunostomy the preferred operative approach.


CT Findings in the External Auditory Canal after Transcanal Surgery.
Middle ear surgery is often performed through the external auditory canal, and the CT appearance of the external auditory canal after transcanal middle ear surgery can mimic erosive pathology such as carcinoma, external auditory canal cholesteatoma, or necrotizing external otitis. We reviewed the CT findings in a group of patients following transcanal surgery to highlight this potential pitfall in interpretation. Twenty-seven temporal bones in 25 patients with a history of a transcanal approach to the middle ear and available postoperative CT imaging were identified. Images were assessed for changes along or involving the walls of the external auditory canal, including widening, irregularity, bony defects, and soft tissue opacification. Osseous changes along the floor of the external auditory canal were demonstrated in 25 of 27 (92.6%) temporal bone CT scans. Similar changes were present in the superior and anterior walls of the external auditory canal in 21 and 18 temporal bones, respectively. The anterior wall was the most common site for complete bony defects (10 of 27 temporal bones). The posterior wall was the least often involved, with osseous changes in 15 of 27 temporal bones and bony defects in 3 cases. Soft tissue thickening was seen most commonly along the floor. No patient was found to have a superimposed pathologic process of the external auditory canal. CT findings in the external auditory canal after transcanal surgery include thinning, irregularity and/or flattening of the bone, soft tissue thickening, and bony wall defects. Although these changes may be subtle, they may mimic pathology and should be included in the differential diagnosis of osseous abnormality of the external auditory canal.


Chloroflexus islandicus sp. nov., a thermophilic filamentous anoxygenic phototrophic bacterium from a geyser.
A novel, thermophilic filamentous anoxygenic phototrophic bacterium, strain isl-2T, was isolated from the Strokkur Geyser, Iceland. Strain isl-2T formed unbranched multicellular filaments with gliding motility. The cells formed no spores and stained Gram-negative. The existence of pili was described in a species of the genus Chloroflexus for the first time, to our knowledge. Optimal growth occurred at a pH range of 7.5-7.7 and at a temperature of 55 °C. Strain isl-2T grew photoheterotrophically under anaerobic conditions in the light and chemoheterotrophically under aerobic conditions in the dark. The major cellular fatty acids were C18 : 1ω9, C16 : 0, C18 : 0 and C18 : 0-OH. The major quinone was menaquinone-10. The photosynthetic pigments were bacteriochlorophylls c and a as well as β- and γ-carotenes. The results of phylogenetic analysis of the 16S rRNA gene sequences placed strain isl-2T into the genus Chloroflexus of the phylum Chloroflexi with Chloroflexus aggregans DSM 9485T as the closest relative (97.0 % identity). The whole-genome sequence of isl-2T was determined. Average nucleotide identity values obtained for isl-2T in comparison to available genomic sequences of other strains of members of the genus Chloroflexus were 81.4 % or less and digital DNA-DNA hybridisation values 22.8 % or less. The results of additional phylogenetic analysis of the PufLM and BchG amino acid sequences supported the separate position of the isl-2T phylotype from the phylotypes of other members of the genus Chloroflexus. On the basis of physiological and phylogenetic data as well as genomic data, it was suggested that isl-2T represents a novel species within the genus Chloroflexus, with the proposed name Chloroflexus islandicus sp. nov. The type strain of the species is isl-2T (=VKM B-2978T,=DSM 29225T,=JCM 30533T).


A tetratricopeptide repeat domain-containing protein SSR1 located in mitochondria is involved in root development and auxin polar transport in Arabidopsis.
Auxin polar transport mediated by a group of Pin-formed (PIN) transporters plays important roles in plant root development. However, the mechanism underlying the PIN expression and targeting in response to different developmental and environmental stimuli is still not fully understood. Here, we report a previously uncharacterized gene SSR1, which encodes a mitochondrial protein with tetratricopeptide repeat (TPR) domains, and show its function in root development in Arabidopsis thaliana. In ssr1-2, a SSR1 knock-out mutant, the primary root growth was dramatically inhibited due to severely impaired cell proliferation and cell elongation. Significantly lowered level of auxin was found in ssr1-2 roots by auxin measurement and was further supported by reduced expression of DR5-driven reporter gene. As a result, the maintenance of the root stem cell niche is compromised in ssr1-2. It is further revealed that the expression level of several PIN proteins, namely, PIN1, PIN2, PIN3, PIN4 and PIN7, were markedly reduced in ssr1-2 roots. In particular, we showed that the reduced protein level of PIN2 on cell membrane in ssr1-2 is due to impaired retrograde trafficking, possibly resulting from a defect in retromer sorting system, which destines PIN2 for degradation in vacuoles. In conclusion, our results indicated that SSR1 is functioning in root development in Arabidopsis, possibly by affecting PIN protein expression and subcellular targeting.


Mitotane (o,p'-DDD) treatment of 200 dogs with pituitary-dependent hyperadrenocorticism.
Two hundred dogs with pituitary dependent hyperadrenocorticism (PDH) were treated with mitotane at an initial daily dosage of 21 to 69 mg/kg (mean = 45.2 mg/kg) for 5 to 14 days. During the induction period, 194 of the dogs also were given daily maintenance dosages of a glucocorticoid. Fifty of the dogs exhibited one or more adverse effects during initial induction, including weakness, vomiting, anorexia, diarrhea, and ataxia. After completion of the induction period, repeat ACTH stimulation testing revealed significant decreases in mean serum cortisol concentrations when compared with initial values. Twenty-five dogs, however, still responded to exogenous ACTH with serum cortisol concentrations above normal resting range, necessitating daily treatment for an additional 5 to 55 days. In contrast, 70 of the 200 dogs had low post-ACTH serum cortisol concentrations after the induction period. These subnormal serum cortisol concentrations generally increased spontaneously to within normal resting range 2 to 6 weeks after cessation of mitotane. In 184 dogs, mitotane was continued at an initial mean maintenance dosage of 49 mg/kg administered weekly in two to three divided doses. Of these dogs, 107 had one or more relapses of hyperadrenocorticism during treatment. In the 75 dogs that had one relapse, the median maintenance dosage was increased by approximately 35%, whereas the median maintenance dosage in the 32 dogs having two or more relapses was eventually increased by 225% over the initial dosage. After a mean maintenance treatment time of 2.0 years, the final maintenance dosage required in the 184 dogs ranged from 26.8 to 330 mg/kg/week.(ABSTRACT TRUNCATED AT 250 WORDS)


Psychotherapy and Artificial Intelligence.
This column explores the challenges involved in providing psychotherapy through artificial intelligence. It reviews artificial intelligence's capacity across schools of therapy to address relevant issues related to privacy, the use of technical interventions, and the therapeutic relationship.


Is smoking associated with alcohol-drug dependence in patients with pain and chronic pain patients? An evidence-based structured review.
The objective of this study was to determine if there is consistent evidence for smoking to be considered a red flag for development of opioid dependence during opioid exposure in patients with pain and chronic pain patients (CPPs). Six hundred and twenty-three references were found that addressed the areas of smoking, pain, and drug-alcohol dependence. Fifteen studies remained after exclusion criteria were applied and sorted into four groupings addressing four hypotheses: patients with pain and CPPs who smoke are more likely than their nonsmoking counterparts to use opioids, require higher opioid doses, be drug-alcohol dependent, and demonstrate aberrant drug-taking behaviors (ADTBs). Each study was characterized by the type of study it represented according to the Agency for Health Care Policy and Research (AHCPR) guidelines and independently rated by two raters according to 13 quality criteria to generate a quality score. The percentage of studies in each grouping supporting/not supporting each hypothesis was calculated. The strength and consistency of the evidence in each grouping was rated by the AHCPR guidelines. In each grouping, 100% of the studies supported the hypothesis for that grouping. The strength and consistency of the evidence was rated as A (consistent multiple studies) for the first hypothesis and as B (generally consistent) for the other. There is limited consistent indirect evidence that smoking status in patients with pain and CPPs is associated with alcohol-drug and opioid dependence. Smoking status could be a red flag for opioid-dependence development on opioid exposure.


Focusing of a Rydberg Positronium Beam with an Ellipsoidal Electrostatic Mirror.
Slow atoms in Rydberg states can exhibit specular reflection from a cylindrical surface upon which an azimuthally periodic potential is imposed. We have constructed a concave mirror of this type, in the shape of a truncated oblate ellipsoid of revolution, which has a focal length of (1.50±0.01) m measured optically. When placed near the center of a long vacuum pipe, this structure brings a beam of n=32 positronium (Ps) atoms to a focus on a position sensitive detector at a distance of (6.03±0.03) m from the Ps source. The intensity at the focus implies an overall reflection efficiency of ∼30%. The focal spot diameter (32±1) mm full width at half maximum is independent of the atoms' flight times from 20 to 60 μs, thus indicating that the mirror is achromatic to a good approximation. Mirrors based on this principle would be of use in a variety of experiments, allowing for improved collection efficiency and tailored transport or imaging of beams of slow Rydberg atoms and molecules.


[Application of computational fluid dynamics based on fluid-structure interaction in aortic dissection].
Aortic dissection is a life-threatening disease with acute onset,rapid progression and high mortality.Hemodynamic factors have important reference value in the development and treatment of aortic dissection.Computational fluid dynamics can intuitively and visually simulate the hemodynamic state of human blood vessels and quantify it numerically.However,computational fluid dynamics without fluid-structure interaction analysis cannot reflect the real situation of the human body.Therefore,the fluid-structure interaction numerical simulation of aortic dissection is worthy of further study to achieve more accurate evaluation of hemodynamic state,postoperative effect and risk prediction.In this paper,the application and research progress of computational fluid dynamics based on fluid-structure interaction technology in aortic dissection are briefly reviewed.


Tuning the spin crossover behavior of the polyanion [(H2O)6Fe3(μ-L)6]6-: the case of the cesium salt.
The magnetic behavior of the polyanion [Fe3(μ-L)6(H2O)6]6- (L2- = (1,2,4-triazol-4-yl)ethanedisulfonate) as the corresponding dimethylammonium salt shows memory effect above room temperature, with a dynamic thermal hysteresis cycle over 90 K and temperature-induced excited spin state trapping (TIESST) phenomena at the highest temperatures reported. Taking advantage of the polyanionic nature of this trimetallic complex, we were able to substitute the dimethylammonium cations by the monovalent heavy alkali metal cesium. This methathesis yielded the salt Cs6[Fe3(μ-L)6(H2O)6], with different molecular packing that increases the number and strength of cation-anion interactions, including a more robust H-bonded network. In this phase, the spin transition still occurs above room temperature, but it is more abrupt and narrow (≈50 K wide hysteresis). Despite these differences, TIESST is observed with almost identical characteristic temperature (TTIESST = 240 K) than in the parent compound, which is an additional experimental evidence supporting the molecular origin of the TIESST behavior in these materials.


[Clinical course of chronic bronchitis in Chernobyl wreckers].
To characterize clinical symptoms of chronic bronchitis (CB) in Chernobyl wreckers. 71 CB male Chernobyl wreckers exposed to low-dose radiation and 43 control patients were examined and treated in a specialized hospital in 1988-1996. Clinical features of CB were investigated by clinical (complaints, history, objective examination), laboratory (total blood count, biochemical and immunological blood picture, coagulogram) and instrumental (fibrobronchoscopy, x-ray, radiometry, spirography) findings with further statistical processing. Chernobyl wreckers more frequently had neurocirculatory asthenia and hemoptysis. The latter was caused by bronchial mucosa atrophy. In chronic non-obstructive bronchitis they exhibited mild bronchoobstruction on the distal level. Laboratory indices showed no essential changes. Chernobyl wreckers have a more severe course of CB.


Exergames for unsupervised balance training at home: A pilot study in healthy older adults.
Exercise videogames (exergames) are gaining popularity as tools for improving balance ability in older adults, yet few exergames are suitable for home-based use. The purpose of the current pilot study was to examine the effects of a 6-week unsupervised home-based exergaming training program on balance performance. Ten community dwelling healthy older adults (age: 75.9 ± 7.2 years) played a newly developed ice skating exergame for six weeks at home. In the game, the speed and direction of a virtual ice skater on a frozen canal were controlled using lateral weight shifts, which were captured using Kinect. Sway characteristics during quiet standing in eyes open (EO), eyes closed (EC) and dual task (DT) conditions were assessed in time and frequency domain before, and after two, four and six weeks of training. Balance was also evaluated using the narrow ridge balance test (NRBT). Multilevel modeling was applied to examine changes in balance ability. Participants played 631 (± 124)min over the intervention period and no subjects dropped out. Balance in terms of sway characteristics improved on average by 17.4% (EO) and 23.3% (EC) after six weeks of training (p<0.05). Differences in rate of improvement (p<0.05) were observed between participants. No intervention effects were found for quiet standing in DT conditions and on the NRBT. In conclusion, the pilot study showed that unsupervised home-based exergaming is feasible in community dwelling older adults, but also that participants do not benefit equally from the program, thereby emphasizing the need for more personalized exergame training programs.


The methionine synthase polymorphism c.2756Aright curved arrow G (D919G) is relevant for disease-free longevity.
The functional polymorphism methionine synthase (MTR) c.2576A-->G (D919G) influences homocysteine and folate metabolism and has been reported to be of protective function against oncological, neurodegenerative and vascular diseases. We analyzed 329 healthy individuals to confirm whether this polymorphism might be of epidemiological impact on disease-free longevity. In our sample, prevalence of the G-allele was significantly higher in the older than in the younger individuals (p=0.005) supporting the thesis that MTR c.2576A-->G is beneficial to disease-free longevity. Separate analysis of female and male subjects revealed that the influence of the MTR genotype on male subjects became relevant at a younger age as opposed to female subjects suggesting a gender-dependent effect.


Cost-minimisation Analysis from a Non-inferiority Trial of Ready-Made versus Custom-Made Spectacles for School Children in India.
Purpose: Uncorrected refractive error is the leading cause of visual impairment in children. Many countries, including India, implement school eye health programmes involving vision screening and provision of free spectacles. This is costly for governments/organisations involved. This analysis estimates potential cost-savings if ready-made spectacles, in addition to traditional custom-made spectacles, are available for dispensing in school eye health programmes.Methods: An economic evaluation was conducted alongside a randomised controlled trial comparing spectacle wear of ready-made spectacles versus custom-made spectacles for children aged 11-15 years in schools in India. A cost-minimisation approach was used to calculate cost-savings of a 'ready-made spectacles available' programme compared with a 'custom-made spectacles only' school programme. The analysis was from a service provider perspective. Main outcomes: cost-saving per child needing spectacles and cost-saving per 1000 children screened.Results: The prevalence of uncorrected refractive error was 2.23%, and 86% of children were eligible for ready-made spectacles. The cost per child needing spectacles in a custom-made spectacles only programme was USD$26.91, and in a ready-made spectacles available programme was $11.15, producing a 58.6% cost-saving per child needing spectacles of $15.76. Considering the total cost of the eye health programme, this equated to a 15.1% cost-saving per 1000 children screened of $361. Results were robust to multivariate sensitivity analyses.Conclusion: Our study is the first to demonstrate the significant cost-saving potential of ready-made spectacles in school eye health programmes for uncorrected refractive error compared with custom-made spectacles alone. This has substantial economic benefits for national/international programmes.


Treatment of uncomplicated hypertension is associated with a reduction in cardiovascular mortality: a Korean national cohort study.
Although the benefit of hypertension treatment is well established in high-risk patients, there is a paucity of evidence regarding the benefit of treatment in patients with uncomplicated hypertension. Hypertensive adult patients were selected from the Korea National Health Insurance Sample Cohort in 2002 and were followed until 2013. Patients with a diagnosis of heart failure, coronary artery disease, stroke, malignancy, diabetes, or chronic kidney disease were excluded. Ultimately, 40 496 patients were divided into three groups: never-treated (N = 6756), treated-from-baseline (N = 28 443), and treated-during-follow-up (N = 5297). Five first-line antihypertensive agents were categorized into four classes: renin-angiotensin system blocker (RASB), beta-blocker, calcium channel blocker (CCB), and diuretics. All-cause mortality, cardiovascular mortality, and hazard ratio were determined. All-cause and cardiovascular mortality rates were significantly lower in both treatment groups than in the never-treated group (all log-rank P < 0.001). Treatment from baseline (hazard ratio = 0.49 for all-cause mortality and hazard ratio = 0.62 for cardiovascular mortality) and treatment started during follow-up (hazard ratio = 0.41 for all-cause mortality and hazard ratio = 0.44 for cardiovascular mortality) were independently associated with lower mortality on multivariable Cox analyses. Although RASB, beta-blocker, and CCB significantly reduced all-cause mortality, multivariable Cox analyses showed that RASB and CCB were closely associated with lower all-cause mortality. In terms of cardiovascular mortality, only CCB was associated with lower cardiovascular mortality on multivariable Cox analyses. Treatment of hypertension significantly reduces mortality in patients with uncomplicated hypertension.


Cloning and nucleotide sequence of the penicillinase antirepressor gene penJ of Bacillus licheniformis.
The penicillinase antirepressor gene, penJ, of Bacillus licheniformis ATCC 9945a was cloned in Escherichia coli by using pMB9 as a vector plasmid. The penicillinase gene, penP, its repressor gene, penI, and penJ were encoded on the cloned 5.2-kilobase HindIII fragment of the recombinant plasmid pTTE71. The penJ open reading frame was composed of 1,803 bases and 601 amino acid residues (molecular weight, 68,388). A Shine-Dalgarno sequence was found 7 bases upstream from the translation start site. Since this sequence was located in the 3'-terminal region of the penI gene, penJ might be transcribed together with penI as a polycistronic mRNA from the penI promoter. Frameshift mutations of penJ were constructed in vitro from pTTE71, and the penJ mutant gene was introduced into B. licheniformis by chromosomal recombination. The transformant B. licheniformis U173 (penP+ penI+ penJ) turned out to be uninducible for penicillinase production, whereas the wild-type strain (penP+ penI+ penJ+) was inducible. Only when these three genes (penP, penI, and PenJ) were simultaneously subcloned in Bacillus subtilis did the plasmid carrier exhibit inducible penicillinase production, as did wild-type B. licheniformis. It was concluded that penJ is involved in the penicillinase induction. The regulation of penP expression by penI and penJ is discussed.


Perspectives on the use of modelling and economic analysis to guide HIV programmes in sub-Saharan Africa.
HIV modelling and economic analyses have had a prominent role in guiding programmatic responses to HIV in sub-Saharan Africa. However, there has been little reflection on how the HIV modelling field might develop in future. HIV modelling should more routinely align with national government and ministry of health priorities, recognising their legitimate mandates and stewardship responsibilities, for HIV and other wider health programmes. Importance should also be placed on ensuring collaboration between modellers, and that joint approaches to addressing modelling questions, becomes the norm rather than the exception. Such an environment can accelerate translation of modelling analyses into policy formulation because areas where models agree can be prioritised for action, whereas areas over which uncertainty prevails can be slated for additional study, data collection, and analysis. HIV modelling should increasingly be integrated with the modelling of health needs beyond HIV, particularly in allocative efficiency analyses, where focusing on one disease over another might lead to worse health overall. Such integration might also enhance partnership with national governments whose mandates extend beyond HIV. Finally, we see a need for there to be substantial and equitable investment in capacity strengthening within African countries, so that African researchers will increasingly be leading modelling exercises. Building a critical mass of expertise, strengthened through external collaboration and knowledge exchange, should be the ultimate goal.


Teff grass for continuous stocking in the Southern High Plains by growing beef steers receiving protein supplements.
This experiment evaluated forage quality, total nutrient yield, water footprint, and growth performance of beef steers receiving protein supplements while grazing Teff grass ['Tiffany'Eragrostis tef (Zucc.) Trotter] over two consecutive growing seasons. Each year, four 2.66-ha irrigated paddocks (experimental units) were stocked with crossbred beef steers (n = 5 per paddock, initial BW = 289 ± 30 for yr 1; and n = 6, initial BW = 286 ± 23 for yr 2) in a randomized complete block design and stocked continuously for 63 d. Daily supplements [0.45 kg/d of cottonseed meal (Control) enough to avoid a negative ruminal N balance; and 0.50% mean paddock BW animal-daily (approximately 1.65 kg) of sorghum-dried distillers grains plus solubles, (DDGS)] were randomly assigned to two paddocks each. Supplement did not influence forage neutral detergent fiber (NDF), acid detergent fiber, crude protein, or in vitro true digestibility (P ≥ 0.54), except for a tendency (P = 0.08) for a numerical increase in NDF content of paddocks with steers that received DDGS supplementation. Paddock nutrient-yields were similar (P ≥ 0.43) between supplement treatments. Supplementation with DDGS produced greater (P = 0.01) cattle shrunk average daily gain (ADG). Predicted teff dry matter intake (DMI), net energy for maintenance (NEm), and growth (NEg) (P ≤ 0.03) were greater with cattle offered Control treatment. Predicted total DMI was similar (P = 0.14) although predicted dietary NEm, NEg, gain:feed, and total BW gain were greater (P ≤ 0.02) with DDGS. Predicted forage intake was greater (P ≤ 0.05) for cattle offered Control treatment. Teff nutrients remaining on d 56 were similar (P = 0.33) between treatments. Water footprint for total production of forage nutrient components did not differ (P ≥ 0.12) by treatments. Nutrient yield and water use efficiency of continuously stocked teff grass was not affected by supplemental regimen. Using DDGS as a supplement may increase BW gain through increased nutrient utilization without hindering teff nutrient production on a continuous stocking system.


Insulin Receptor Isoforms in Physiology and Disease: An Updated View.
The insulin receptor (IR) gene undergoes differential splicing that generates two IR isoforms, IR-A and IR-B. The physiological roles of IR isoforms are incompletely understood and appear to be determined by their different binding affinities for insulin-like growth factors (IGFs), particularly for IGF-2. Predominant roles of IR-A in prenatal growth and development and of IR-B in metabolic regulation are well established. However, emerging evidence indicates that the differential expression of IR isoforms may also help explain the diversification of insulin and IGF signaling and actions in various organs and tissues by involving not only different ligand-binding affinities but also different membrane partitioning and trafficking and possibly different abilities to interact with a variety of molecular partners. Of note, dysregulation of the IR-A/IR-B ratio is associated with insulin resistance, aging, and increased proliferative activity of normal and neoplastic tissues and appears to sustain detrimental effects. This review discusses novel information that has generated remarkable progress in our understanding of the physiology of IR isoforms and their role in disease. We also focus on novel IR ligands and modulators that should now be considered as an important strategy for better and safer treatment of diabetes and cancer and possibly other IR-related diseases.


[Diagnostic capability of PULSAR, FDT y HRT-II in glaucoma suspects].
To determine the diagnostic capability of PULSAR-T30W, FDT-Threshold-N30 and HRT-II in glaucoma suspects. Forty-seven eyes from 47 referred glaucoma suspects (GS) were examined twice with each technique. Cases with TOP-WW-MD>6dB were excluded. Results were compared with those of 70 eyes from 70 normal controls (C). Mean MD value using TOP-WW in the GS group (0.96dB. sd=1.7) was not significantly different from C (0.8dB. sd=1.77) (p>0.05). Disc area in GS group (2.12 mm(2). sd=0.34) was significantly greater than in C (1.97 mm2. sd=0.45) (p<0.01). For 95.7% specificity, PULSAR-sLV showed the highest sensitivity of 30.9% in individual examinations. The highest reproducible sensitivity in the two examinations was obtained using HRT-II maximum contour elevation (23.4%) and reference height (23.4%), and was 14.9% for various indices after correcting for the influence of disc area (cup area, cup/disc area ratio, maximum contour depression and mean RNFL thickness). Reproducible sensitivity of the perimetric indices was: PULSAR-MD=8.5%, PULSAR-sLV=17%, FDT-MD=6.4%, FDT-PSD=4.3%. The association of perimetric and HRT-II indices achieved high sensitivity but low diagnostic reproducibility. The most effective indices were maximum contour elevation, reference height and PULSAR-sLV, although the inclusion of the optic nerve head assessment in the selection of the GS sample may have favored the HRT-II results.


Expression of γ-tocopherol methyltransferase in chloroplasts results in massive proliferation of the inner envelope membrane and decreases susceptibility to salt and metal-induced oxidative stresses by reducing reactive oxygen species.
The γ-tocopherol methyltransferase (γ-TMT) is an important enzyme regulating synthesis of four tocopherols (α, γ, β and δ). In this report, we investigated the role of γ-TMT in regulating abiotic stress within chloroplasts. The At γ-tmt overexpressed via the tobacco chloroplast genome accumulated up to 7.7% of the total leaf protein, resulting in massive proliferation of the inner envelope membrane (IEM, up to eight layers). Such high-level expression of γ-TMT converted most of γ-tocopherol to α-tocopherol in transplastomic seeds (~10-fold higher) in the absence of abiotic stress. When grown in 400 mm NaCl, α-tocopherol content in transplastomic TMT leaves increased up to 8.2-fold and 2.4-fold higher than wild-type leaves. Likewise, under heavy metal stress, α-tocopherol content in the TMT leaves increased up to 7.5-fold, twice higher than in the wild type. Under extreme salt stress, the wild type accumulated higher starch and total soluble sugars, but TMT plants were able to regulate sugar transport. Hydrogen peroxide and superoxide content in wild type increased up to 3-fold within 48 h of NaCl stress when compared to TMT plants. The ion leakage from TMT leaves was significantly less than wild-type plants under abiotic stress and with less malondialdehyde, indicating lower lipid peroxidation. Taken together, these studies show that α-tocopherol plays a crucial role in the alleviation of salt and heavy metal stresses by decreasing ROS, lipid peroxidation and ion leakage, in addition to enhancing vitamin E conversion. Increased proliferation of the IEM should facilitate studies on retrograde signalling from chloroplast to the nucleus.


The Role of Autophagy, Mitophagy and Lysosomal Functions in Modulating Bioenergetics and Survival in the Context of Redox and Proteotoxic Damage: Implications for Neurodegenerative Diseases.
Redox and proteotoxic stress contributes to age-dependent accumulation of dysfunctional mitochondria and protein aggregates, and is associated with neurodegeneration. The free radical theory of aging inspired many studies using reactive species scavengers such as alpha-tocopherol, ascorbate and coenzyme Q to suppress the initiation of oxidative stress. However, clinical trials have had limited success in the treatment of neurodegenerative diseases. We ascribe this to the emerging literature which suggests that the oxidative stress hypothesis does not encompass the role of reactive species in cell signaling and therefore the interception with reactive species with antioxidant supplementation may result in disruption of redox signaling. In addition, the accumulation of redox modified proteins or organelles cannot be reversed by oxidant intercepting antioxidants and must then be removed by alternative mechanisms. We have proposed that autophagy serves this essential function in removing damaged or dysfunctional proteins and organelles thus preserving neuronal function and survival. In this review, we will highlight observations regarding the impact of autophagy regulation on cellular bioenergetics and survival in response to reactive species or reactive species generating compounds, and in response to proteotoxic stress.


Molecular, immunohistochemical, and pharmacological evidence of oxytocin's role as inhibitor of carbohydrate but not fat intake.
Oxytocin (OT) facilitates feeding termination stemming from high osmolality, stomach distention, and malaise. Recent knockout (KO) studies suggested a crucial function for OT in carbohydrate intake: OT-/- mice had increased preference for carbohydrates, including sucrose, but not fat (Intralipid). In striking contrast, sugar appetite was unaffected in the OT receptor KO mouse; data from wild-type animals have been insufficient. Therefore, we examined the involvement of OT in the regulation of sucrose vs. fat intake in C57BL/6 mice that served as a background KO strain. We exposed mice to a meal of sucrose or Intralipid and determined that the percentage of c-Fos-immunoreactive paraventricular hypothalamic OT neurons was elevated at termination of intake of either of the tastants, but this increase was 2-fold higher in sucrose-fed mice. A 48-h exposure to sucrose compared with Intralipid caused up-regulation of OT mRNA, whereas inherent individual preferences for sucrose vs. fat were not associated with differences in baseline OT expression as established with quantitative PCR. We found that L-368,899, an OT receptor antagonist, increased sugar intake when sucrose was presented alone or concurrently with Intralipid; it had no effect on Intralipid or total calorie consumption. L-368,899 affected Fos immunoreactivity in the paraventricular hypothalamus, arcuate nucleus, amygdala, and nucleus of the solitary tract, areas involved in aversion, satiety, and reward. This pattern serves as neuroanatomical basis of OT's complex role in food intake, including sucrose intake. The current findings expand our knowledge on OT and suggest that it acts as a carbohydrate-specific inhibitor of feeding.


Multiple rapid automatic naming measures of cognition: normal performance and effects of aging.
Rapid automatic naming tasks are clinical tools for probing brain functions that underlie normal cognition. To compare performance for various stimuli in normal subjects and assess the effect of aging, we administered six single-dimension stimuli (color, form, number, letter, animal, and object) and five dual-dimension stimuli (color-form, color-number, color-letter, color-animal, and color-object) to 144 normal volunteers who ranged in age from 15 to 85 years. Rapid automatic naming times for letters and numbers were significantly less than for forms, animals, and objects. Rapid automatic naming times for color-number and color-letter stimuli were significantly less than for color-form, color-animal, or color-object stimuli. Age correlated significantly with rapid automatic naming time for each single-dimension stimulus and for color-form, color-number, color-animal, and color-object stimuli. Linear regression showed that rapid automatic naming times increased with age for aggregated color stimuli, aggregated single-dimension stimuli, and aggregated dual-dimension stimuli. This age effect persisted in subgroups less than 60 years of age and greater than 60 years of age. We conclude that normal performance time is dependent on the task, with letter and number stimuli eliciting most rapid responses, and that most rapid automatic naming times increase with age.


Determination of Dermal Pharmacokinetics by Microdialysis Sampling in Rats.
The evaluation of absorption and availability at the site of action of a drug candidate is an important element of drug discovery and development, as clinical response is a function of the bioavailability of the active agent and its continued presence at the site of action. Evaluation of dermal pharmacokinetics facilitates the selection of new compounds or chemical structures for advancement as possible clinical candidates. An advantage of microdialysis is that it allows the measurement of compound concentrations at the site of action without disturbing the tissue milieu, making it possible to determine the relationship between this important variable and plasma concentrations of the agent. Described in this unit are laboratory protocols for performing dermal microdialysis experiments in rat for the purpose of defining the pharmacokinetics parameters of test agents. © 2019 by John Wiley & Sons, Inc.


Pigment-dispersing factor is involved in age-dependent rhythm changes in Drosophila melanogaster.
Most animals show rest/activity rhythms that are regulated by an endogenous timing mechanism, the so-called circadian system. The rhythm becomes weaker with age, but the mechanism underlying the age-associated rhythm change remains to be elucidated. Here we employed Drosophila melanogaster as a model organism to study the aging effects on the rhythm. We first investigated activity rhythms under light-dark (LD) cycles and constant darkness (DD) in young (1-day-old) and middle-aged (30-, 40-, and 50-day-old) wild-type male flies. The middle-aged flies showed a reduced activity level in comparison with young flies. Additionally, the free-running period significantly lengthened in DD, and the rhythm strength was diminished. Immunohistochemistry against pigment-dispersing factor (PDF), a principal neurotransmitter of the Drosophila clock, revealed that PDF levels declined with age. We also found an attenuation of TIMELESS (TIM) oscillation in the cerebral clock neurons in elder flies. Intriguingly, overexpression of PDF suppressed age-associated changes not only in the period and strength of free-running locomotor rhythms but also in the amplitude of TIM oscillations in many pacemaker neurons in the elder flies, suggesting that the age-dependent PDF decline is responsible for the rhythm attenuation. These results suggest that the age-associated reduction of PDF may cause attenuation of intercellular communication in the circadian neuronal network and of TIM cycling, which may result in the age-related rhythm decay.


Absence of Helicobacter pylori in pediatric adenoid hyperplasia.
To (1) develop a reverse transcription-polymerase chain reaction assay to determine whether Helicobacter pylori and/or other members of the Helicobacteraceae family are detected in hyperplastic adenoids of children and (2) critically analyze published polymerase chain reaction methods to ascertain whether false-positive detection of H pylori has been reported. Cohort study. Adenoid biopsy specimens (78 hyperplastic and 15 normal) were collected from children aged 2 to 10 years. Total RNA was extracted before reverse transcription of bacterial RNA using Helicobacteraceae-specific primer. A nested reverse transcription-polymerase chain reaction protocol was designed to detect all species of the Helicobacteraceae family. A piece of each biopsy specimen was examined histologically. Laryngopharyngeal reflux was suspected in 41% of the children (n = 23) on the basis of the Reflux Symptom Index. No evidence of H pylori was found in any adenoid sample. Candidatus Wolinella africanus was the only Helicobacteraceae family member detected in 1 hyperplastic adenoid. Histologic examination identified very few bacterial organisms. Previous polymerase chain reaction findings may be the result of false-positive H pylori detection. Inflammation and enlargement of the adenoids is not likely due to ongoing bacterial infection arising from laryngopharyngeal reflux. We conclude that H pylori and other Helicobacteraceae family members are not major contributors to the development of hyperplastic adenoids in children.


Comparison of reports on lung function tests made by chest physicians with those made by a simple computer program.
The numerical results of 60 sets of pulmonary function tests were submitted to five consultant chest physicians for independent reporting. The chest physicians' reports were compared with the reports generated by the on-line computer reporting system in routine use in the pulmonary function laboratory at Westminster Hospital. There was good agreement between the reports of the chest physicians among themselves and with the computer. The individual reports were compared with the consensus opinions of the physicians and the computer. The computer's decisions differed from the consensus opinion in 4.6% of instances. The comparable figures for the five physicians were respectively 2.9%, 2.9%, 2.9%, 4.0%, and 4.6%. Decisions differing from the consensus were due to mistakes or actual sustained disagreements of opinion. The physicians made more mistakes than the computer even though they were performing under test conditions. They made up for this, however, by producing fewer genuine disagreements from the consensus opinions. It is concluded that in routine day-to-day practice the computer report will be as consistently useful as the chest physicians' reports and more immediately available.


Design of a Wearable Fingertip Haptic Device for Remote Palpation: Characterisation and Interface with a Virtual Environment.
This paper presents the development of a wearable Fingertip Haptic Device (FHD) that can provide cutaneous feedback via a Variable Compliance Platform (VCP). The FHD includes an inertial measurement unit, which tracks the motion of the user's finger while its haptic functionality relies on two parameters: pressure in the VCP and its linear displacement towards the fingertip. The combination of these two features results in various conditions of the FHD, which emulate the remote object or surface stiffness properties. Such a device can be used in tele-operation, including virtual reality applications, where rendering the level of stiffness of different physical or virtual materials could provide a more realistic haptic perception to the user. The FHD stiffness representation is characterised in terms of resulting pressure and force applied to the fingertip created through the relationship of the two functional parameters - pressure and displacement of the VCP. The FHD was tested in a series of user studies to assess its potential to create a user perception of the object's variable stiffness. The viability of the FHD as a haptic device has been further confirmed by interfacing the users with a virtual environment. The developed virtual environment task required the users to follow a virtual path, identify objects of different hardness on the path and navigate away from "no-go" zones. The task was performed with and without the use of the variable compliance on the FHD. The results showed improved performance with the presence of the variable compliance provided by the FHD in all assessed categories and particularly in the ability to identify correctly between objects of different hardness.


The selection of grammatical features in word production: the case of plural nouns in German.
Two experiments investigate the effect of number congruency using picture-word interference. Native German participants were required to name pictures of single objects (Nase 'nose') or two instances of the same object (Nasen 'noses') while ignoring simultaneously presented distractor words. Distractor words either had the same number or were different in number. In addition, the type of plural formation (same or different inflectional plural suffix) and the semantic relationship (same or different semantic category) between target and distractor were varied in Experiments 1 and 2. Results showed no effect of number congruency in either experiment. Furthermore, the type of inflectional suffix did not exert an influence on naming latencies in Experiment 1, but semantic relationship led to a significant interference effect in Experiment 2. The results indicate that selection of the number feature diacritic in noun production is not a competitive process. The implications of the results for models of lexical access are discussed.


Laparoscopic surgical box model training for surgical trainees with no prior laparoscopic experience.
Surgical training has traditionally been one of apprenticeship, where the surgical trainee learns to perform surgery under the supervision of a trained surgeon. This is time consuming, costly, and of variable effectiveness. Training using a box model physical simulator - either a video box or a mirrored box - is an option to supplement standard training. However, the impact of this modality on trainees with no prior laparoscopic experience is unknown. To compare the benefits and harms of box model training versus no training, another box model, animal model, or cadaveric model training for surgical trainees with no prior laparoscopic experience. We searched the Cochrane Central Register of Controlled Trials (CENTRAL), MEDLINE, EMBASE, and Science Citation Index Expanded to May 2013. We included all randomised clinical trials comparing box model trainers versus no training in surgical trainees with no prior laparoscopic experience. We also included trials comparing different methods of box model training. Two authors independently identified trials and collected data. We analysed the data with both the fixed-effect and the random-effects models using Review Manager for analysis. For each outcome, we calculated the standardised mean difference (SMD) with 95% confidence intervals (CI) based on intention-to-treat analysis whenever possible. Twenty-five trials contributed data to the quantitative synthesis in this review. All but one trial were at high risk of bias. Overall, 16 trials (464 participants) provided data for meta-analysis of box training (248 participants) versus no supplementary training (216 participants). All the 16 trials in this comparison used video trainers. Overall, 14 trials (382 participants) provided data for quantitative comparison of different methods of box training. There were no trials comparing box model training versus animal model or cadaveric model training. Box model training versus no training: The meta-analysis showed that the time taken for task completion was significantly shorter in the box trainer group than the control group (8 trials; 249 participants; SMD -0.48 seconds; 95% CI -0.74 to -0.22). Compared with the control group, the box trainer group also had lower error score (3 trials; 69 participants; SMD -0.69; 95% CI -1.21 to -0.17), better accuracy score (3 trials; 73 participants; SMD 0.67; 95% CI 0.18 to 1.17), and better composite performance scores (SMD 0.65; 95% CI 0.42 to 0.88). Three trials reported movement distance but could not be meta-analysed as they were not in a format for meta-analysis. There was significantly lower movement distance in the box model training compared with no training in one trial, and there were no significant differences in the movement distance between the two groups in the other two trials. None of the remaining secondary outcomes such as mortality and morbidity were reported in the trials when animal models were used for assessment of training, error in movements, and trainee satisfaction. Different methods of box training: One trial (36 participants) found significantly shorter time taken to complete the task when box training was performed using a simple cardboard box trainer compared with the standard pelvic trainer (SMD -3.79 seconds; 95% CI -4.92 to -2.65). There was no significant difference in the time taken to complete the task in the remaining three comparisons (reverse alignment versus forward alignment box training; box trainer suturing versus box trainer drills; and single incision versus multiport box model training). There were no significant differences in the error score between the two groups in any of the comparisons (box trainer suturing versus box trainer drills; single incision versus multiport box model training; Z-maze box training versus U-maze box training). The only trial that reported accuracy score found significantly higher accuracy score with Z-maze box training than U-maze box training (1 trial; 16 participants; SMD 1.55; 95% CI 0.39 to 2.71). One trial (36 participants) found significantly higher composite score with simple cardboard box trainer compared with conventional pelvic trainer (SMD 0.87; 95% CI 0.19 to 1.56). Another trial (22 participants) found significantly higher composite score with reverse alignment compared with forward alignment box training (SMD 1.82; 95% CI 0.79 to 2.84). There were no significant differences in the composite score between the intervention and control groups in any of the remaining comparisons. None of the secondary outcomes were adequately reported in the trials. The results of this review are threatened by both risks of systematic errors (bias) and risks of random errors (play of chance). Laparoscopic box model training appears to improve technical skills compared with no training in trainees with no previous laparoscopic experience. The impacts of this decreased time on patients and healthcare funders in terms of improved outcomes or decreased costs are unknown. There appears to be no significant differences in the improvement of technical skills between different methods of box model training. Further well-designed trials of low risk of bias and random errors are necessary. Such trials should assess the impacts of box model training on surgical skills in both the short and long term, as well as clinical outcomes when the trainee becomes competent to operate on patients.


Transition between superhydrophobic states on rough surfaces.
Surface roughness is known to amplify hydrophobicity. It is observed that, in general, two drop shapes are possible on a given rough surface. These two cases correspond to the Wenzel (liquid wets the grooves of the rough surface) and Cassie (the drop sits on top of the peaks of the rough surface) formulas. Depending on the geometric parameters of the substrate, one of these two cases has lower energy. It is not guaranteed, though, that a drop will always exist in the lower energy state; rather, the state in which a drop will settle depends typically on how the drop is formed. In this paper, we investigate the transition of a drop from one state to another. In particular, we are interested in the transition of a "Cassie drop" to a "Wenzel drop", since it has implications on the design of superhydrophobic rough surfaces. We propose a methodology, based on energy balance, to determine whether a transition from the Cassie to Wenzel case is possible.


RGS10 physically and functionally interacts with STIM2 and requires store-operated calcium entry to regulate pro-inflammatory gene expression in microglia.
Chronic activation of microglia is a driving factor in the progression of neuroinflammatory diseases, and mechanisms that regulate microglial inflammatory signaling are potential targets for novel therapeutics. Regulator of G protein Signaling 10 is the most abundant RGS protein in microglia, where it suppresses inflammatory gene expression and reduces microglia-mediated neurotoxicity. In particular, microglial RGS10 downregulates the expression of pro-inflammatory mediators including cyclooxygenase 2 (COX-2) following stimulation with lipopolysaccharide (LPS). However, the mechanism by which RGS10 affects inflammatory signaling is unknown and is independent of its canonical G protein targeted mechanism. Here, we sought to identify non-canonical RGS10 interacting partners that mediate its anti-inflammatory mechanism. Through RGS10 co-immunoprecipitation coupled with mass spectrometry, we identified STIM2, an endoplasmic reticulum (ER) localized calcium sensor and a component of the store-operated calcium entry (SOCE) machinery, as a novel RGS10 interacting protein in microglia. Direct immunoprecipitation experiments confirmed RGS10-STIM2 interaction in multiple microglia and macrophage cell lines, as well as in primary cells, with no interaction observed with the homologue STIM1. We further determined that STIM2, Orai channels, and the calcium-dependent phosphatase calcineurin are essential for LPS-induced COX-2 production in microglia, and this pathway is required for the inhibitory effect of RGS10 on COX-2. Additionally, our data demonstrated that RGS10 suppresses SOCE triggered by ER calcium depletion and that ER calcium depletion, which induces SOCE, amplifies pro-inflammatory genes. In addition to COX-2, we also show that RGS10 suppresses the expression of pro-inflammatory cytokines in microglia in response to thrombin and LPS stimulation, and all of these effects require SOCE. Collectively, the physical and functional links between RGS10 and STIM2 suggest a complex regulatory network connecting RGS10, SOCE, and pro-inflammatory gene expression in microglia, with broad implications in the pathogenesis and treatment of chronic neuroinflammation.


Predictors of sexual desire disorders in women.
A historic belief was that testosterone was the "hormone of desire." However, recent data, which show either minimal or no significant correlation between testosterone levels and women's sexual desire, suggest that nonhormonal variables may play a key role. To compare women with hypoactive sexual desire disorder (HSDD) and those with the recently proposed more symptomatic desire disorder, Sexual Desire/Interest Disorder (SDID), on the relative contribution of hormonal vs. nonhormonal variables. Women with HSDD (N = 58, mean age 52.5) or SDID (N = 52, mean age 50.9) participated in a biopsychosocial assessment in which six nonhormonal domains were evaluated for the degree of involvement in the current low desire complaints. Participants provided a serum sample of hormones analyzed by gas chromatography-mass spectrometry or liquid chromatography/mass spectrometry/mass spectrometry. Logistic regression was used to assess the ability of variables (nonhormonal: history of sexual abuse, developmental history, psychosexual history, psychiatric status, medical history, and sexual/relationship-related factors; hormonal: dehydroepiandrosterone [DHEA], 5-diol, 4-dione, testosterone, 5-α-dihydrotestosterone, androsterone glucuronide, 3α-diol-3G, 3α-diol-17G, and DHEA-S; and demographic: age, relationship length) to predict group membership. Women with SDID had significantly lower sexual desire and arousal scores, but the groups did not differ on relationship satisfaction or mood. Addition of the hormonal variables to the two demographic variables (age, relationship length) did not significantly increase predictive capability. However, the addition of the six nonhormonal variables to these two sets of predictors significantly increased ability to predict group status. Developmental history, psychiatric history, and psychosexual history added significantly to the predictive capability provided by the basic model when examined individually. Nonhormonal variables added significant predictive capability to the basic model, highlighting the importance of their assessment clinically where women commonly have SDID in addition to HSDD, and emphasizing the importance of addressing psychological factors in treatment.


Cell volume regulation and signaling in 3T3-L1 pre-adipocytes and adipocytes: on the possible roles of caveolae, insulin receptors, FAK and ERK1/2.
Caveolae have been implicated in sensing of cell volume perturbations, yet evidence is still limited and findings contradictory. Here, we investigated the possible role of caveolae in cell volume regulation and volume sensitive signaling in an adipocyte system with high (3T3-L1 adipocytes); intermediate (3T3-L1 pre-adipocytes); and low (cholesterol-depleted 3T3-L1 pre-adipocytes) caveolae levels. Using large-angle light scattering, we show that compared to pre-adipocytes, differentiated adipocytes exhibit several-fold increased rates of volume restoration following osmotic cell swelling (RVD) and osmotic cell shrinkage (RVI), accompanied by increased swelling-activated taurine efflux. However, caveolin-1 distribution was not detectably altered after osmotic swelling or shrinkage, and caveolae integrity, as studied by cholesterol depletion or expression of dominant negative Cav-1, was not required for either RVD or RVI in pre-adipocytes. The insulin receptor (InsR) localizes to caveolae and its expression dramatically increases upon adipocyte differentiation. In pre-adipocytes, InsR and its effectors focal adhesion kinase (FAK) and extracellular signal regulated kinase (ERK1/2) localized to focal adhesions and were activated by a 5 min exposure to insulin (100 nM). Osmotic shrinkage transiently inhibited InsR Y(146)-phosphorylation, followed by an increase at t=15 min; a similar pattern was seen for ERK1/2 and FAK, in a manner unaffected by cholesterol depletion. In contrast, cell swelling had no detectable effect on InsR, yet increased ERK1/2 phosphorylation. In conclusion, differentiated 3T3-L1 adipocytes exhibit greatly accelerated RVD and RVI responses and increased swelling-activated taurine efflux compared to pre-adipocytes. Furthermore, in pre-adipocytes, Cav-1/caveolae integrity is not required for volume regulation. Given the relationship between hyperosmotic stress and insulin signaling, the finding that cell volume regulation is dramatically altered upon adipocyte differentiation may be relevant for the understanding of insulin resistance and metabolic syndrome.


Reconstruction of small and fragile bile ducts without mucosa-to-mucosa anastomosis.
We describe a simple and easy technique for performing choledochojejunostomy without the need to suture the full thickness of the ductal and intestinal walls for patients in whom standard choledochojejunostomy is difficult because the stumps of the residual bile ducts are small and fragile. This technique is useful in partial liver transplantation or after hepatectomy that includes removal of the extrahepatic bile ducts. The procedure involves the placing of external biliary drainage tubes through a Roux-en-Y jejunal loop, positioned transanastomotically, and the use of an external jejunostomy to decompress the loop. The tubes are fixed to the jejunal loop by a purse-string suture and to the duct by simple ligation or a purse-string suture. Anastomosis is performed by suturing the connective tissue and liver parenchyma around the ductal stump to the seromuscular layer of the intestine. Choledochojejunostomy according to this method was performed in 5 cases; the biliary drainage tubes were removed 1 to 4 months after surgery. The only complications were cerebellar infarction and cholangitis, both of which resolved with conservative treatment. We consider that this technique will be helpful as a last-ditch measure when standard choledochojejunostomy, with suturing of the full thickness of the walls of the duct and intestine to secure mucosa-to-mucosa apposition, is impossible because of small and fragile bile ducts.


The Usefulness of Duplex Ultrasound for Hemodialysis Access Selection.
A native vessel is preferable to an artificial graft for dialysis access. Duplex ultrasound (DUS) is noninvasive, cost-effective modality to evaluate the vessels for dialysis. The purpose of this study was to compare the rates of utilization of native vessels after preoperative imaging with DUS and contrast venography (CV). A retrospective review was performed on patients who received an arteriovenous fistula (AVF) or arteriovenous graft (AVG) between June 2006 and July 2010. Patients were classified into 3 groups. In group 1, CV was used to evaluate the vessel. Both DUS and CV were used in group 2. In group 3, only DUS was used. The frequency of utilization of a native vessel was analyzed in each group. The chi-square test was used for statistical analysis. During the study period, 173 patients received an AVF or AVG. Eighty-nine patients were male. The mean age was 60.6±14.6 years. A native vessel was used in 56/81 patients (69.1%) and 74/81 patients (91.4%) in groups 1 and 3, respectively (P<0.001). In group 2, all patients underwent access procedures using native vessels. AVG was initially planned for 2 patients in group 2 after vessel evaluation using CV, but a native vessel was successfully used because DUS identified optimal vessels for AVF. The 1-year primary patency rate was similar in 3 groups. Preoperative DUS is safe and easy to use for vessel evaluation, and can be used as a primary imaging modality for creation of access.


Seed desiccation mechanisms co-opted for vegetative desiccation in the resurrection grass Oropetium thomaeum.
Resurrection plants desiccate during periods of prolonged drought stress, then resume normal cellular metabolism upon water availability. Desiccation tolerance has multiple origins in flowering plants, and it likely evolved through rewiring seed desiccation pathways. Oropetium thomaeum is an emerging model for extreme drought tolerance, and its genome, which is the smallest among surveyed grasses, was recently sequenced. Combining RNA-seq, targeted metabolite analysis and comparative genomics, we show evidence for co-option of seed-specific pathways during vegetative desiccation. Desiccation-related gene co-expression clusters are enriched in functions related to seed development including several seed-specific transcription factors. Across the metabolic network, pathways involved in programmed cell death inhibition, ABA signalling and others are activated during dehydration. Oleosins and oil bodies that typically function in seed storage are highly abundant in desiccated leaves and may function for membrane stability and storage. Orthologs to seed-specific LEA proteins from rice and maize have neofunctionalized in Oropetium with high expression during desiccation. Accumulation of sucrose, raffinose and stachyose in drying leaves mirrors sugar accumulation patterns in maturing seeds. Together, these results connect vegetative desiccation with existing seed desiccation and drought responsive pathways and provide some key candidate genes for engineering improved drought tolerance in crop plants.


Correlation of the virus sequence content and biological properties of cells carrying the herpes simplex virus type 2 thymidine kinase gene.
Seven cell lines, transformed to a thymidine kinase-positive phenotype with herpes simplex virus type 2 (HSV-2) DNA fragments, have been examined to determine their virus-specific DNA sequence content. The results are consistent with the reported map position of the HSV-2 thymidine kinase gene. Different cell lines contained different amounts of non-selected virus-specific sequences and this correlated with the ability of some cell lines to compensate the deficiencies in some temperature-sensitive (ts) mutants of HSV-1 and HSV-2. The cell lines were also examined for their ability to synthesize elevated levels of thymidine kinase in response to infection with a thymidine kinase-negative virus mutant. Of the seven cell lines, two failed to respond to infection in this way and these cell lines lacked sequences on the upstream side of the kinase gene which were present in the remaining cell lines.


A prospective observational study of decisional capacity determinations in an academic medical center.
The main goals, of this observational study were to clarify and categorize decisional capacity assessments performed by a psychosomatic medicine (PSM) consult service at an academic medical center. Areas of investigation included types of decisional capacity requests, the relationship between Folstein Mini-Mental State Exam (MMSE) and PSM decisional capacity determination, psychiatric diagnoses of patients, and the agreement between primary clinician capacity assessment and that of the PSM clinician. This was a prospective study of 100 consecutive decisional capacity consultations. Twelve requests were for second consultations for patients previously seen, for a net of 88 patients. In 77 cases, patients lacked decisional capacity. Assessments for capacity to leave against medical advice (AMA), capacity to accept medical/surgical procedures, capacity to refuse medical/surgical procedures, and capacity to participate in discharge planning were nearly equally frequent. An MMSE cutoff score of < 21 was found to be 100% specific and 69% sensitive for identifying the patients without capacity, while a cutoff of < 24 was found to be 83% sensitive and 90% specific for identifying patients without capacity. The primary psychiatric diagnosis was a cognitive disorder in 52 cases. In 38 of the 39 cases where the primary team believed the patient lacked capacity, the PSM service agreed. In only two of the seven cases where primary teams believed patients had capacity did the PSM team agree. The MMSE was a useful predictor of capacity determination by PSM personnel, but using MMSE alone results in a number of erroneous determinations. Cognitive disorders were the most common primary psychiatric diagnoses in decisional capacity cases. Primary teams tended to be more accurate when they found patients to lack capacity.


beta-cell function in children with diabetes.
Although it is generally believed that insulin secretion is minimal or absent in juvenile-onset diabetes, we have found appreciable levels of C-peptide at the time of onset in 12 patients, 4 to 16 years old (9.3 +/- 4.2). Ten of them had ketonuria but none severe ketoacidosis. All entered a remission period. Most of the patients had near normal C-peptide levels during the remission, and their beta cells had the capacity to respond to a breakfast stimulation with increased insulin secretion. C-peptide and proinsulin were also determined in 98 juvenile diabetics with age at onset of 1 to 16 years (6.8 +/- 3.9) and a duration of diabetes between two and 17 years (6.7 +/- 3.4). Many were found to have persisting beta-cell function, which seems to be of importance for ensuring stability in metabolic control. Although little is known about factors that may slow or reverse the process leading to beta-cell failure, our results suggest that early detection and intensive treatment of diabetes before severe metabolic disturbances have occurred may help preserve beta-cell function.


20 years of leptin: role of leptin in human reproductive disorders.
Leptin, as a key hormone in energy homeostasis, regulates neuroendocrine function, including reproduction. It has a permissive role in the initiation of puberty and maintenance of the hypothalamic-pituitary-gonadal axis. This is notable in patients with either congenital or acquired leptin deficiency from a state of chronic energy insufficiency. Hypothalamic amenorrhea is the best-studied, with clinical trials confirming a causative role of leptin in hypogonadotropic hypogonadism. Implications of leptin deficiency have also emerged in the pathophysiology of hypogonadism in type 1 diabetes. At the other end of the spectrum, hyperleptinemia may play a role in hypogonadism associated with obesity, polycystic ovarian syndrome, and type 2 diabetes. In these conditions of energy excess, mechanisms of reproductive dysfunction include central leptin resistance as well as direct effects at the gonadal level. Thus, reproductive dysfunction due to energy imbalance at both ends can be linked to leptin.