Gender, violence and brief interventions for alcohol in the emergency department.
The impact of gender and violence on brief interventions (BIs) for alcohol use in the emergency department (ED) has not been studied. Our objective was to examine the effectiveness of alcohol BIs in an ED population stratified by gender and violence. This was a secondary analysis of datasets pooled from three ED-based randomized controlled studies of alcohol BIs. AUDIT-C was the primary outcome measure; secondary outcomes were binge drinking and achievement of NIAAA safe drinking levels. We conducted univariate comparisons and developed generalized linear models (GLM) for the primary outcome and generalized estimating equation (GEE) models for secondary outcomes to examine the intervention effect on the whole study group, gender-stratified subgroups, and gender- and violence-stratified subgroups. Of 1219 participants enrolled, 30% were female; 31% of women and 42% of men reported violence involvement at baseline. In univariate analysis, no differences in outcomes were found between intervention and control groups for any subgroup. However, in multivariable models, men demonstrated an intervention effect for likelihood of safe drinking limits. Stratifying further by violence, only men without violence involvement demonstrated a positive intervention effect for safe drinking limits. There was no evidence of an intervention effect on women. Analyzing the overall effect of ED-based BI may mask its ability to improve alcohol-related outcomes in a subset of the population. Alternatively, interventions may need to be significantly improved in subsets of the ED population, e.g., in women and in men with involvement in violence.


Identifying and quantifying apoptosis: navigating technical pitfalls.
Apoptosis or programmed cell death is often altered in malignancies and is frequently determined by the terminal transferase-mediated nick end labeling technique (TUNEL). However, commercially available protocols can produce high background and false-positive staining, which renders the distinction between apoptosis and necrosis difficult. In an attempt to develop a rapid and reproducible method for detecting and quantifying apoptosis, we coupled optimization of the Apoptag Plus Peroxidase In Situ Apoptosis Detection kit with quantitative histomorphometric computer imaging software using the Bacus Laboratories Incorporated Slide Scanner (BLISS). Multiple (200-350) unique 40x images were scanned using the BLISS system and downloaded into the WebSlide Browser program, creating a permanent, scanned record of the area assessed. The stored images were counted, with the final analysis simultaneously taking into account cells that were immunohistochemically positive and the histology of the surrounding cells to reduce the possibility of false positive and negative staining. In addition, cells with equivocal staining can be simultaneously reviewed by other technologists with networked WebSlide Browser access to the same images. Our data show that the advantages offered by the BLISS imaging software greatly reduce the potential drawbacks of using the TUNEL method as a sole means of quantification.


Trimethoprim-sulfamethoxazole-induced hepatotoxicity in a pediatric patient.
Due to the escalating rates of community-acquired methicillin-resistant Staphylococcus aureus (CA-MRSA) infection, trimethoprim-sulfamethoxazole (TMP-SMX) is being used increasingly in the pediatric population for skin and soft tissue infections. Although this combination agent has been associated with a hypersensitivity syndrome involving cutaneous skin eruptions, pediatric cases of TMP-SMX-induced hepatotoxicity are rare. We describe a relatively healthy, 9-year-old boy who developed a CA-MRSA skin and soft tissue infection and was treated with TMP-SMX. After 14 days of therapy, he was taken to the emergency department with a 3-day history of fever, headache, and neck pain. He was diagnosed with a viral syndrome, acetaminophen was prescribed, and he was sent home. Three days later, the patient returned to the emergency department with fever, vomiting, decreased energy and appetitie, and suprapubic abdominal pain, and he was hospitalized. Laboratory test results revealed elevated liver function test values. After other potential causes of liver toxicity were excluded, TMP-SMX was determined to be the cause of his acute liver toxicity. The drug was discontinued, his symptoms resolved, and his liver function tests returned to normal. Use of the Naranjo adverse drug reaction probability scale indicated a probable relationship (score of 5) between the patient's development of hepatotoxicity and the TMP-SMX therapy. This rare adverse reaction to TMP-SMX has been reported in adults; however, to our knowledge, it has been reported in only five other children. Due to the increasing use of TMP-SMX in children, clinicians should be aware of this potentially life-threatening, immunemediated hypersensitivity reaction. Fortunately, however, the hepatotoxicity appears to resolve after discontinuation of the TMP-SMX therapy in most reported cases. This case report illustrates the importance of early detection of drug-induced hepatotoxicity and timely drug discontinuation to prevent the need for liver transplantation.


Intracellular cytokine and cathelicidin secretion from monocytes and neutrophils in childhood tuberculosis.
Human cathelicidin (hCAP 18) is one of the main immunomodulators in the local immune response to Mycobacterium tuberculosis. There has been no information regarding the role of hCAP 18+ in the in vivo production of cytokine in childhood pulmonary tuberculosis (TB). We aimed to determine the intracellular cytokine secretion, including hCAP 18+ from monocytes and neutrophils of pulmonary TB in children, compared with healthy children. Fifteen patients with pulmonary TB were enrolled in the study as the study group, and 15 healthy children as the control group, between the age of 1 and 16 years. The patients' 25 hydroxyvitamin D levels were measured. The expression of hCAP 18+, TNF α, INF-γ and IL-8 from CD14+ monocytes and CD15 + neutrophils was analyzed using the flow cytometry method. The statistical analysis was performed with PASW Statistics v.13.0. The mean vitamin D level was similar in both groups (P = 0.78). The expression of hCAP 18+ (P = 0.0001) and IL-8 from CD14 + (P = 0.0001) monocytes were significantly higher in the study group compared with the control group. There was no difference in the hCAP 18+ and IL-8 expression in CD 15+ neutrophils in both groups. The expression of TNF α, INF-γ from CD14+ monocytes and CD 15+ neutrophils, in both study and control groups revealed no statistical differences.The level of hCAP 18+ and IL-8 released from monocytes were enhanced in the serum in childhood pulmonary TB. The present study is the first report detecting the intracellular hCAP 18+ expression in vivo in childhood pulmonary TB.


A qualitative analysis of semantic intrusive errors in Alzheimer's disease.
Intrusive or perseverative errors as evidenced on tests of memory have been found to occur with greater frequency in patients with Alzheimer's Disease (AD) relative to other disorders and are thought to reflect the cholinergic deficits associated with AD. A particular type of intrusive error, semantic intrusions, has been found to discriminate AD from multi-infarct dementia and normal elderly controls. However, it has been suggested that such errors actually reflect random responding as the result of the disinhibition often observed in AD patients. The present study compared the frequency and types of intrusive errors from a sample of mildly, moderately, and more severely impaired AD patients who had undergone the Fuld Object Memory Evaluation with the "guesses" of elderly controls. Results of this study indicate that the semantic intrusions of AD patients differ both in type and frequency from the guesses of the control subjects.


Fuchs endothelial corneal dystrophy in patients with myotonic dystrophy: a case series.
The aim was to report 4 cases of Fuchs endothelial corneal dystrophy (FECD) in patients with an established diagnosis of myotonic dystrophy (DM) and suggest a mechanism for their association based on the known molecular genetics and potential pathophysiological parallels of DM and FECD. We reviewed all available medical records and pathology slides for the 4 reported cases from the Department of Ophthalmology at Oregon Health and Science University's Casey Eye Institute and Devers Eye Institute at the Legacy Good Samaritan Medical Center in Portland, OR. Four patients were found to have DM and bilateral corneal guttae, consistent with the diagnosis of FECD. All the identified patients were female and were aged between 34 and 63, and 2 patients were related (mother and daughter). The corneal specimens from 2 of the 4 patients who had undergone a corneal transplant were pathologically confirmed to be consistent with the diagnosis of FECD. To our knowledge, FECD has not been previously reported in association with DM. Because both diseases are somewhat prevalent in the United States, it is possible that their coexistence is merely a coincidence in these patients. However, recent studies into the pathogenesis of each disease have shown more parallels between FECD and DM, suggesting the possibility of a noncoincidental association. Potential mutual pathogenic mechanisms may involve altered protein expression causing the deregulation of ion homeostasis, an unstable intronic trinucleotide repeat expansion, or activation of the unfolded protein response and oxidative stress pathways.


Sonographic and electrophysiologic findings in patients with meralgia paresthetica.
This study was aimed to demonstrate the usefulness of ultrasonography in the diagnosis of meralgia paresthetica (MP). Clinical data, sensory nerve conduction study and sonographic findings of 23 patients with unilateral MP were analyzed retrospectively. Twelve healthy subjects were recruited for the optimal cut-off value for the cross-sectional area of LFCN to diagnose MP using receiver operating characteristic curves. The median value of the cross-sectional area of LFCN in healthy subjects was 3mm(2) and in the affected and unaffected sides in patients with MP, they were 11 and 6.5mm(2) around the anterior superior iliac spine level. The optimal cut-off value for the diagnosis of MP was 5mm(2). The location of LFCN on the affected side was superior to the inguinal ligament in 12 out of 23 cases (50%). Ultrasonography is useful in the diagnosis of MP as a supplemental diagnostic tool, which gives important information about the morphologic changes that occur in the LFCN and its course. To the best of our knowledge, this is the first report to suggest that describe the cutoff value of the cross-sectional area of LFCN for the diagnosis of MP using ultrasonography.


Association of Non-host Crop Plants with Mandarin in Host Location and Survival of Diaphorina citri Kuwayama (Hemiptera: Psyllidae).
Research efforts have been made to develop novel tactics, such as those targeting behavioral control, for management of the Asian citrus psyllid Diaphorina citri Kuwayama (Hemiptera: Psyllidae), vector of the causal agent of citrus Huanglongbing. Here, we investigated whether association of "Ponkan" mandarin (Citrus reticulata) with volatiles from non-host crops: avocado, passion fruit or coffee, alters host location by the Asian citrus psyllid; and whether they can be temporary hosts for the Asian citrus psyllid. In wind tunnel assays, we found that the association of mandarin seedling with avocado plant volatiles reduced in 30% the number of psyllids sitting on host plants compared to the mandarin alone. In contrast, passion fruit plant volatiles facilitated host location by psyllids, which found mandarin seedlings faster than when exposed to mandarin alone. The association with coffee volatiles did not alter the attractiveness of mandarin to the Asian citrus psyllid. Survival and half-lethal time (LT50) of D. citri fed on non-host plants were longer than those insects with water only, but shorter than those fed on mandarin. Among the non-host plants, D. citri performed better in coffee, followed by avocado and passion fruit plants. Our results indicate that the association of mandarin with avocado plant can be beneficial for Asian citrus psyllid management.


A Survey of the Opinion and Experience of UK Dentists: Part 2: Risk Assessment Strategies and the Management of Iatrogenic Trigeminal Nerve Injuries Related to Dental Implant Surgery.
Dental implant-related iatrogenic injuries are proportionally increasing with dental implant surgery. This study assessed the experience of implant-related trigeminal nerve (TG) injuries among UK dentists. Risk management strategies and management of implant-related inferior alveolar nerve (IAN), mental nerve (MN), and lingual nerve injuries were investigated. A survey was distributed among 405 dentists attending an Association of Dental Implantology (ADI) congress, of which 187 completed the survey. Most dentists (76% of 134 responses) allowed a 2 to 4 mm safety zone radiologically above the IAN when placing implants, and over half of the responders (56%) used implants that were 10 mm in length. The most frequent precautionary measure used by 73 (80%) responders was antibiotic coverage routinely to reduce the risk of infection when placing grafts in the posterior mandible. Other precautionary measures included unilateral staging of implant placement (57%), and 43% always identified the MN when placing implants. Nineteen dentists used steroids (eg, dexamethasone) routinely preoperatively and postoperatively. Twenty-six dentists used basic cone-beam CT (CBCT) minimally invasive techniques, and drill stops during implant placement were used by 14 responders. Although it is not highly recommended, steroids were used to manage the neuropathic pain and discomfort experienced by patients with IAN injuries in 40% of cases. Further training of dentists undertaking implant surgery is required so that they acquire up-to-date and evidence-based knowledge and skills in the prevention, diagnosis, and management of dental implant-related TG injuries. This training should also involve the justification and interpretation of CBCTs.


Efficacy of rotator cuff suture and arthroscopic 360° capsular release in patients with rotator cuff tear with limited shoulder movement.
To determine the clinical efficacy of rotator cuff suture and arthroscopic 360° capsular release in patients with rotator cuff tendinopathy to improve the Constant-Murley and Visual Analogue Scale (VAS) scores, and shoulder flexion. Fifty-one patients with full-thickness rotator cuff tears and limited shoulder movement who were admitted to our hospital from October 2017 to October 2020 were selected; all patients were treated with arthroscopic rotator cuff suture and 360° capsular release. The Constant-Murley score, VAS score, and shoulder flexion angle were used to evaluate shoulder joint function before and during follow-up. Rotator cuff healing was assessed by MRI with the Sugaya classification. After treatment, the Constant-Murley score (58.98 ± 9.84) was significantly improved compared with pre-treatment (29.33 ± 9.71), the VAS score (1.23 ± 0.87) was significantly lower than pre-treatment (7.54 ± 1.22), and the shoulder flexion angle (142.67 ± 8.59°) was significantly improved compared with pre-treatment (51.50 ± 2.10°); the difference was statistically significant (P < 0.05). Arthroscopic rotator cuff suture and simultaneous 360° capsular release have a significant effect on the treatment of rotator cuff tear with limited shoulder movement.


Neonatal size of low socio-economic status Black and White term births in Albany County, NYS.
Birth weight has long been a focus of study by epidemiologists and human biologists, because it reflects the quality of the intrauterine environment and may be used as a predictor of future growth and development. Comparisons of Black and White neonates in the USA have consistently shown differences in birth weight. Confounding variables are a major problem in any such investigation, especially socio-economic status which is highly correlated with race in the USA. This study was distinctive in the sampling of one socio-economic stratum (low income), and the use of five anthropometric measures in addition to birth weight. The goals of this study were as follows: to determine if there were differences in body size and body composition at birth in Black and White neonates of low socio-economic status (SES), and to investigate what variables might account for any observed variability. The sample consisted of full term Black and White neonates of low SES (n = 323) born in Albany, NY (1986-1997). Birth weight, length, head and arm circumference, and subscapular and triceps skinfolds were compared. Race was determined through maternal self-identification. White neonates were significantly larger than Black neonates in birth weight, length and head circumference. Among female neonates none of the anthropometric dimensions differed between Blacks and Whites. Among male neonates, Whites were significantly larger than Blacks in birth weight, length, head and arm circumferences. Principal components analysis reduced the six anthropometric dimensions to two summary measures: body size and composition. When controlling for social and biological variables, race and sex were significant predictors of body composition, but not body size. Interpretation of results and possible causal relationships are discussed.


Progesterone and interferon-tau regulate cystatin C in the endometrium.
Cystatin C (CST3) is a secreted inhibitor of lysosomal cysteine proteases cathepsins B (CTSB) and CTSL, which are abundant in the ovine endometrium and conceptus. In mice, cathepsins and cystatins play important roles in implantation and placentation. This study determined effects of the estrous cycle, pregnancy, progesterone (P4), and interferon-tau (IFNT) on CST3 in the ovine uterus. In cyclic ewes, CST3 mRNA was low on d 10, increased about 12-fold by d 12, and declined thereafter. In early pregnant ewes, CST3 mRNA was low on d 10 and increased about 130-fold from d 10 to d 20. CST3 mRNA and protein were abundant in the endometrial luminal epithelium (LE) and glandular epithelium and also in conceptus trophectoderm. In uterine flushes from pregnant ewes, CST3 protein was not detected on d 10 but was abundant on d 12, 14, and 16. In another study, treatment of ovariectomized, cyclic ewes with P4 induced a 14-fold increase in endometrial CST3 mRNA, and IFNT stimulated an additional 2-fold increase in CST3 mRNA in P4-treated ewes but not in ewes treated with P4 and the antiprogestin ZK 136,317. CST3 mRNA and protein were abundant in the endometrial luminal epithelium and superficial glandular epithelium of P4-treated ewes but were very low or not detectable in endometria of P4- and ZK-treated ewes. These results indicate that CST3 is a novel P4-induced and IFNT-stimulated gene expressed only in the epithelial cells of the ovine endometrium and implicate CST3 in regulation of uterine cathepsin activity during conceptus implantation.


Cln5 represents a new type of cysteine-based S-depalmitoylase linked to neurodegeneration.
Genetic CLN5 variants are associated with childhood neurodegeneration and Alzheimer's disease; however, the molecular function of ceroid lipofuscinosis neuronal protein 5 (Cln5) is unknown. We solved the Cln5 crystal structure and identified a region homologous to the catalytic domain of members of the N1pC/P60 superfamily of papain-like enzymes. However, we observed no protease activity for Cln5; and instead, we discovered that Cln5 and structurally related PPPDE1 and PPPDE2 have efficient cysteine palmitoyl thioesterase (S-depalmitoylation) activity using fluorescent substrates. Mutational analysis revealed that the predicted catalytic residues histidine-166 and cysteine-280 are critical for Cln5 thioesterase activity, uncovering a new cysteine-based catalytic mechanism for S-depalmitoylation enzymes. Last, we found that Cln5-deficient neuronal progenitor cells showed reduced thioesterase activity, confirming live cell function of Cln5 in setting S-depalmitoylation levels. Our results provide new insight into the function of Cln5, emphasize the importance of S-depalmitoylation in neuronal homeostasis, and disclose a new, unexpected enzymatic function for the N1pC/P60 superfamily of proteins.


[Study of toxicity of denture prosthetic appliance prothyl hot on the Jurkat cell model system].
The aim of our study was the investigation of toxicity of denture prosthetic appliance Prothyl Hot on the model system of Jurkat cell culture. As a result of our study it was revealed that denture prosthetic appliance Prothyl Hot: - hadn't manifested toxic effects the viability Jurkat cells (that reviled by stability of activity of mitochondrial dehydrogenises and mean of mitochondrial membrane potential; - didn't influence on the balance of pro-and antyinflammatory cytokines, expressed by Jurkat cells; - induces intensification of oxidative metabolism in Jurkat cells, which may be considered as compensatory reaction developing in the cells.


RasGRP1 overexpression in the epidermis of transgenic mice contributes to tumor progression during multistage skin carcinogenesis.
RasGRP1 is a guanine nucleotide exchange factor for Ras, activated in response to the second messenger diacylglycerol and its ultrapotent analogues, the phorbol esters. We have previously shown that RasGRP1 is expressed in mouse epidermal keratinocytes and that transgenic mice overexpressing RasGRP1 in the epidermis under the keratin 5 promoter (K5.RasGRP1) are prone to developing spontaneous papillomas and squamous cell carcinomas, suggesting a role for RasGRP1 in skin tumorigenesis. Here, we examined the response of the K5.RasGRP1 mice to multistage skin carcinogenesis, using 7,12-dimethylbenz(a)anthracene as carcinogen and 12-O-tetradecanoylphorbol-13-acetate (TPA) as tumor promoter. We found that whereas tumor multiplicity did not differ between transgenic and wild-type groups, the transgenic tumors were significantly larger than those observed in the wild-type mice (wild-type, 4.58 +/- 0.25 mm; transgenic, 9.83 +/- 1.05 mm). Histologic analysis further revealed that squamous cell carcinomas generated in the transgenic mice were less differentiated and more invasive than the wild-type tumors. Additionally, 30% of the transgenic mice developed tumors in the absence of initiation, suggesting that RasGRP1 overexpression could partially substitute for the initiation step induced by dimethylbenz(a)anthracene. In primary keratinocytes isolated from K5.RasGRP1 mice, TPA stimulation induced higher levels of Ras activation compared with the levels measured in the wild-type cells, indicating that constitutive overexpression of RasGRP1 in epidermal cells leads to elevated biochemical activation of endogenous Ras in response to TPA. The present data suggests that RasGRP1 participates in skin carcinogenesis via biochemical activation of endogenous wild-type Ras and predisposes to malignant progression in cooperation with Ras oncogenic signals.


Clinical impact of intesified 5-Fluorouracil-based chemotherapy using a prospective pharmacokinetically-guided dosing approach: comparative study in elderly and non-elderly patients with metastatic colorectal cancer.
We compared clinical outcome and pharmacokinetic (pK) parameters of a new pharmacokinetically-guided dosing strategy in two groups of patients (age < or >65 years) with metastatic colorectal cancer (mCRC). We retrospectively analyzed all the patients with mCRC, receiving standard LV5FU2 regimen during cycle-1, intensified from cycle-2 onwards according to an adaptation schedule based on the area under the concentration-time curve (AUC) value (mg.h/l.m(2)) of 5-FU measured during cycle-1. Among the 103 eligible patients, the 48 elderly (median age 70 range 65-80) did not differ significantly from the 55 non-elderly patients (median age 59 range 33-64) in pPK parameters (including AUC at cycle-1 and cycle-2), efficacy (objective response rate of 27% [16.1-40.9] in younger and 35% [22.2-50.5] in older patients, p = 0.4) and tolerability (33.3% of overall grade 3-4 toxicities in older patients and 34.5% in younger, p = 0.9). Our data indicated high dose-intensity values, not significantly different between elderly and non-elderly patients. it was feasible to increase the 5-fU continuous infusion dose in 43/55 (78%) younger and 40/48 (83%) older patients (p=0.509) and even to double it in half of the patients of both groups. Aging did not seem to limit intensified chemotherapy or to affect the pK behavior of the 5-FU.


Visuoconstructional drawing ability in the differential diagnosis of neurologic compromise versus depression.
Neuropsychologists are often called upon to assist in making differential diagnoses between neurologic compromise and depression. However, the literature has been inconsistent about the impact of depression on neurocognitive functioning. This makes interpretation of neuropsychological test data among depressed individuals difficult. In a retrospective cross-sectional study of 200 patients, the relationship between depression (Beck Depression Inventory or Geriatric Depression Scale) and visuoconstructional drawing ability/praxis (Rey-Osterrieth Copy [RO]) was evaluated. Age and education both correlated significantly with RO performance (p <.001 and.005, respectively). There were no significant relationships found between copy performance and depression, even when controlling for the significant effects of age and education. A prospective longitudinal study of 20 psychiatric inpatients with depression tested on admission and again at discharge matched with 20 nondepressed normal controls yielded similar results in that the two groups did not differ in their initial RO performance. Moreover, an improvement in depression as measured by a decrease in Beck Depression Inventory scores did not coincide with any improvement in RO scores. By comparison, a matched positive control group of 20 neurologically compromised patients was significantly worse than both the normal control and the depressed groups on the RO (p <.05). These data suggest that poor performance on the RO should not be interpreted simply as a consequence of depression. Age and education, though, should be considered when interpreting the RO.


Misoprostol moistened with acetic acid or saline for second trimester pregnancy termination: a randomized prospective double-blind trial.
This study was conducted to evaluate the efficacy and side effects of a new regimen of 800 microg misoprostol administered intravaginally every 6 h up to a maximum of three doses in 24 h for second trimester pregnancy termination. A total of 66 women seeking termination of second trimester pregnancy (30 fetal structural anomaly, six chromosomal abnormality and 30 fetal death) were randomly assigned to one of two treatment groups: (i) intravaginal misoprostol moistened with 3 ml of 5% acetic acid in group A (n = 33); or (ii) intravaginal misoprostol moistened with 3 ml of saline in group B (n = 33). The overall median (range) induction-abortion interval was 10 h (2-46) [10 h (4-35) in 36 live fetuses and 9 h (2-46) in 30 dead fetuses, P = 0.515]. All of the patients in both groups aborted within 48 h (100% success rate). The median (range) induction-abortion interval revealed a significantly faster delivery time (P < 0.001) in group A [8 h (2-24)] than in group B [14 h (3-46)]. This new regimen of 800 microg of vaginal misoprostol every 6 h for a maximum of three doses in 24 h was an effective alternative method for second trimester abortion. In addition, misoprostol moistened with acetic acid was significantly more effective than misoprostol moistened with saline.


Amelioration of Diabetes in a Murine Model upon Transplantation of Pancreatic β-Cells with Optogenetic Control of Cyclic Adenosine Monophosphate.
Pharmacological augmentation of glucose-stimulated insulin secretion (GSIS), for example, to overcome insulin resistance in type 2 diabetes is linked to suboptimal regulation of blood sugar. Cultured β-cells and islets expressing a photoactivatable adenylyl cyclase (PAC) are amenable to GSIS potentiation with light. However, whether PAC-mediated enhancement of GSIS can improve the diabetic state remains unknown. To this end, β-cells were engineered with stable PAC expression that led to over 2-fold greater GSIS upon exposure to blue light while there were no changes in the absence of glucose. Moreover, the rate of oxygen consumption was unaltered despite the photoinduced elevation of GSIS. Transplantation of these cells into streptozotocin-treated mice resulted in improved glucose tolerance, lower hyperglycemia, and higher plasma insulin when subjected to illumination. Embedding optogenetic networks in β-cells for physiologically relevant control of GSIS will enable novel solutions potentially overcoming the shortcomings of current treatments for diabetes.


Dexamethasone effects on rat hepatocyte spheroid formation and function.
Hepatocytes cultured on moderately adhesive surfaces or in spinner flasks spontaneously self-assemble into spherical tissue-like aggregates (spheroids). These spheroids have smooth surfaces and tissue-like polarized cell morphology, including bile canalicular-like channels, and maintain high viability and liver-specific functions for extended culture periods. Dexamethasone (DEX), a synthetic glucocorticoid, is known to elicit various responses in gene expression, and is often added to hepatocyte culture medium. The morphology and liver-specific protein production of hepatocyte spheroids were assessed under DEX concentrations ranging from 50 nM to 10 microM. DEX altered the kinetics of spheroid formation in a concentration-dependent fashion, with increasing concentrations inhibiting aggregation and promoting aggregate disassembly on culture dishes. DEX addition to spinner cultures resulted in smaller, more irregularly shaped spheroids and a higher incidence of aggregate clumping. Albumin and urea production were also higher in DEX cultures, but this effect was not as sensitive to concentration and occurred irrespective of the state of aggregation. RTPCR was utilized to assess the mRNA levels of extracellular matrix proteins, E-cadherin, and cytochrome P-450 enzymes. Results indicated a slight increase in fibronectin and collagen III mRNA early in the cultures, possibly contributing to the changes in morphology observed.


CODEX2: full-spectrum copy number variation detection by high-throughput DNA sequencing.
High-throughput DNA sequencing enables detection of copy number variations (CNVs) on the genome-wide scale with finer resolution compared to array-based methods but suffers from biases and artifacts that lead to false discoveries and low sensitivity. We describe CODEX2, as a statistical framework for full-spectrum CNV profiling that is sensitive for variants with both common and rare population frequencies and that is applicable to study designs with and without negative control samples. We demonstrate and evaluate CODEX2 on whole-exome and targeted sequencing data, where biases are the most prominent. CODEX2 outperforms existing methods and, in particular, significantly improves sensitivity for common CNVs.


Evidence for adrenergic control of transcellular calcium distribution in liver.
Free Ca2+ concentration and 45Ca flux were measured in the perfusate and bile of the perfused rat liver. With a perfusate Ca2+ concentration of 1 mM, the bile concentration was 0.35 mM. The ratio of 45Ca in bile to that in blood increased from 0.3 to 0.6 over 90 min of perfusion. Both verapamil and adrenaline (via alpha-adrenergic receptors) increased the 45Ca bile/perfusate ratio to 0.8. Adrenaline infusion increased the bile Ca2+ concentration to 0.8 mM. This decreased to 0.35 mM after the infusion was stopped.


Injury of the spino-thalamo-cortical pathway is necessary for central post-stroke pain.
We investigated the relationship between injury of the spino-thalamo-cortical pathway (STP) and central post-stroke pain (CPSP) in patients with intracerebral hemorrhage, using diffusion tensor tractography (DTT). 30 consecutive chronic patients, in whom integrity of the STP and the medial lemnisco-thalamo-cortical pathway (MLP) were spared in both hemispheres, were recruited. We classified the patients into two groups according to the presence of the CPSP. DTTs were obtained using the FMRIB Software Library. Fractional anisotropy (FA), mean diffusivity (MD), and tract volume were measured. A laterality index (LI) was used to determine asymmetry of DTT parameters between the hemispheres. The LI for tract volume of the STP in the CPSP group was lower than that of the non-CPSP group (p = 0.000). However, there were no differences between the CPSP group and non-CPSP group for the LIs of FA or MD of the STP or for the LIs of FA, MD, or tract volume of the MLP (p > 0.05). Decrease of tract volume without changes in the STP FA or MD values in the CPSP group indicates partial injury of STP. Therefore, injury of the STP seems to be a requirement for the development of CPSP in patients with intracerebral hemorrhage.


Experimental study and clinical observations of autologous hair follicle cell transplants to treat stable vitiligo.
Vitiligo is characterized by the loss and/or dysfunction of melanocytes in the skin and has a profound impact on the social interactions of patients. Although there are many treatment options for vitiligo, the outcome is frequently unsatisfactory, especially for patients with stable vitiligo. To study the biological properties of melanocytes derived from human hair follicles and to observe the efficacy of using transplants of autologous hair follicle cells to treat patients with stable vitiligo. From February 2014 to March 2017, 26 patients with stable vitiligo, who were refractory to all current routine therapy, underwent treatment with transplanted autologous hair follicle cells. The skin graft from each patient's occipital region was trimmed to remove excess adipose tissue and some of the upper part of the dermis. The remaining tissue, including hair follicles and dermal papillae, was cut into pieces and incubated in collagenase type IV and then in trypsin-ethylenediaminetetraacetic acid solutions. The cells were recovered, resuspended in the patient's own serum and then applied to the recipient area. Clinical observations continued for 6 months to 1 year. Laboratory experiments were also performed during this time on scalp specimens obtained from normal human volunteers. Cells migrating from the outer root sheath and the dermal papillae at various times of culture were observed using a microscope. Most of the repigmentation in the vitiligo areas appeared within 8 weeks of transplantation of autologous hair follicle cells. Early skin repigmentation was not uniform and appeared more repigmented than the surrounding normal skin. As time went by, the repigmentation became more obvious and matched the color of the skin around the lesion. Most of the pigmentation presented as a diffuse pattern and was not localized around the hair follicles. Among the 26 patients, 9 (34.6%) achieved excellent repigmentation, while 13 (50.0%) had good, 3 (11.5%) fair and 1 (3.9%) poor repigmentation. During the follow-up visit at 1 year, no excess hair growth was observed in the recipient areas and there was no scarring or ulcer formation in the donor or recipient areas. In the experimental part of the study, many keratinocytes, melanocytes and fibroblasts migrated from the adherent outer root sheath. In later subcultures using a specialized medium, pure melanocytes were obtained that had a strong proliferative capacity and had bipolar or poly-dendritic shapes. On the other hand, cells from the dermal papillae grew radially in primary culture and were almost fibroblast-like. However, a few bipolar melanocytes appeared in the later stage of culture. The results of our study show that transplantation of autologous hair follicle cells is a simple and effective method to treat patients with stable vitiligo. Hair follicles (especially the outer root sheath) harbor many melanocytes with potential proliferative ability. There are a few limitations of the present study: a small sample size, a short follow-up period, no cell counting or viability testing.


Food poisoning and Staphylococcus aureus enterotoxins.
Staphylococcus aureus produces a wide variety of toxins including staphylococcal enterotoxins (SEs; SEA to SEE, SEG to SEI, SER to SET) with demonstrated emetic activity, and staphylococcal-like (SEl) proteins, which are not emetic in a primate model (SElL and SElQ) or have yet to be tested (SElJ, SElK, SElM to SElP, SElU, SElU2 and SElV). SEs and SEls have been traditionally subdivided into classical (SEA to SEE) and new (SEG to SElU2) types. All possess superantigenic activity and are encoded by accessory genetic elements, including plasmids, prophages, pathogenicity islands, vSa genomic islands, or by genes located next to the staphylococcal cassette chromosome (SCC) implicated in methicillin resistance. SEs are a major cause of food poisoning, which typically occurs after ingestion of different foods, particularly processed meat and dairy products, contaminated with S. aureus by improper handling and subsequent storage at elevated temperatures. Symptoms are of rapid onset and include nausea and violent vomiting, with or without diarrhea. The illness is usually self-limiting and only occasionally it is severe enough to warrant hospitalization. SEA is the most common cause of staphylococcal food poisoning worldwide, but the involvement of other classical SEs has been also demonstrated. Of the new SE/SEls, only SEH have clearly been associated with food poisoning. However, genes encoding novel SEs as well as SEls with untested emetic activity are widely represented in S. aureus, and their role in pathogenesis may be underestimated.


Kohlschütter-Tönz Syndrome - Report of an additional case.
Kohlschütter-Tönz Syndrome is a rare disorder clinically characterized by amelogenesis imperfecta, epilepsy and progressive mental deterioration. We present an additional case of this syndrome of a nine year-old boy who was referred by pigmented teeth. The mental deterioration was associated with speech delay, impulsive behavior, attention-deficit/hyperactivity disorder, and learning problems. The physical examination revealed a reduction of lower third, slightly palpebral fissures, low ear and hair implantation, coarse hair and hypertrichosis. The intraoral examination showed alteration in teeth pigmentation diagnosed as amelogenesis imperfecta. Although rare, the present case report illustrates a syndrome that has dental anomalies and systemic alterations. It is important to recognize this syndrome as early as possible and paediatric dentist may contribute to the diagnosis and consequently to better manage the patients. Key words:Kohlschütter-Tönz syndrome, amelogenesis imperfecta, seizures, mental deterioration.


Single dose paracetamol (acetaminophen), with and without codeine, for postoperative pain.
Patient surveys have shown that postoperative pain is often not managed well, and there is a need to assess the efficacy and safety of commonly used analgesics as newer treatments become available. Paracetamol (acetaminophen) is an important non-opiate analgesic, commonly prescribed, as well as being available for retail sale. This review seeks to examine the efficacy of paracetamol alone and in combination with codeine, and also considers adverse effects. To assess the analgesic efficacy and adverse effects of a single dose of oral paracetamol (acetaminophen) alone and in combination with codeine for moderate to severe postoperative pain. Published trials were identified from: Medline (1966 to May 1996), Embase (1980 to 1996), Cochrane Library (Issue 2 1996) and the Oxford Pain Relief Database (1950 to 1994). Additional trials were identified from reference lists of retrieved studies. Date of most recent searches: July 1998. Inclusion criteria were: full journal publication, postoperative pain, postoperative oral administration, adult patients, baseline pain of moderate to severe intensity, double-blind design, and random allocation to treatment groups which compared paracetamol with placebo or a combination of paracetamol and codeine with either placebo or the same dose of paracetamol alone. Data were extracted by two independent reviewers, and trials were quality scored. Summed pain intensity and pain relief data were extracted and converted into dichotomous information to yield the number of patients with at least 50% pain relief. This was used to calculate the relative benefit and number-needed-to-treat (NNT) for one patient to achieve at least 50% pain relief over 4 to 6 hours compared with placebo. Adverse effects were used to calculate relative risk and number-needed-to-harm (NNH). We found 40 trials of paracetamol against placebo (4171 patients), 22 trials of paracetamol plus codeine against placebo (1407 patients) and 12 trials of paracetamol plus codeine against the same dose of paracetamol (794 patients). In postoperative pain paracetamol 1000 mg had an NNT of 4.6 (3.8-5.4) for at least 50% pain relief when compared with placebo, and paracetamol 600/650 mg had an NNT of 5.3 (4.1-7.2). Paracetamol 600/650 mg plus codeine 60 mg had an NNT of 3. 6 (2.9-4.5). Comparing paracetamol plus codeine 60 mg with the same dose of paracetamol alone gave an NNT of 7.7 (5.1-17) for at least 50% pain relief. Adverse effects: Relative risk estimates for paracetamol 600/650 mg plus codeine 60 mg versus placebo showed a significant difference for 'drowsiness'/somnolence (NNH 11 (7.5- 0)) and dizziness (NNH 27 (15-164)) but no significant difference for nausea/vomiting. Paracetamol is an effective analgesic with a low incidence of adverse effects. The addition of codeine 60 mg to paracetamol produces additional pain relief even in single oral doses, but may be accompanied by an increase in drowsiness and dizziness.


Creation of an Online Platform for Identification of Microorganisms: Peak Picking or Full-Spectrum Analysis.
Identification of microorganisms by MALDI-TOF mass spectrometry is a very efficient method with high throughput, speed, and accuracy. However, it is significantly limited by the absence of a universal database of reference mass spectra. This problem can be solved by creating an Internet platform for open databases of protein spectra of microorganisms. Choosing the optimal mathematical apparatus is the pivotal issue for this task. In our previous study we proposed the geometric approach for processing mass spectrometry data, which represented a mass spectrum as a vector in a multidimensional Euclidean space. This algorithm was implemented in a Jacob4 stand-alone package. We demonstrated its efficiency in delimiting two closely related species of the Bacillus pumilus group. In this study, the geometric approach was realized as R scripts which allowed us to design a Web-based application. We also studied the possibility of using full spectra analysis (FSA) without calculating mass peaks (PPA), which is the logical development of the method. We used 74 microbial strains from the collections of ICiG SB RAS, UNIQEM, IEGM, KMM, and VGM as the models. We demonstrated that the algorithms based on peak-picking and analysis of complete data have accuracy no less than that of Biotyper 3.1 software. We proposed a method for calculating cut-off thresholds based on averaged intraspecific distances. The resulting database, raw data, and the set of R scripts are available online at https://icg-test.mydisk.nsc.ru/s/qj6cfZg57g6qwzN.


Endoscopy of the spinal cord: cadaveric study and clinical experience.
Recent improvements in instruments permit endoscopic examination of previously inaccessible sites. We report on the clinical use of a small-diameter endoscope to examine the spinal subarachnoid space, cord surface and syrinx cavities. Prior to clinical application, three types of endoscopes with external diameters of 0.5, 1.4 or 2.2 mm were inserted percutaneously in the lumbar region of five cadavers for preclinical evaluation of the procedure and the three endoscopes. The observations permitted us to perform spinal endoscopy preoperatively or intraoperatively using the 0.5-mm instrument in seven patients with spinal cord lesions between 1995 and 1997. The patients included two with spinal cord herniation through a dural defect, two with syringomyelia, one with spinal arachnoid cyst, one with spinal epidural cyst and one undergoing lumboperitoneal shunt for hydrocephalus. In patients in whom an endoscope was used preoperatively, the endoscope provided morphological information useful in preoperative diagnosis and planning surgical strategy. When the endoscope was used intraoperatively, areas outside the field of vision of a microscope could be examined, and physiological evaluation could include visualizing improved cord perfusion from the spinal subarachnoid space after surgery. Endoscopes could be safely inserted and approached to the lesions under direct vision while avoiding blood vessels and nerve roots on the spinal cord surface. No changes in symptoms or complications occurred in association with endoscopy. Using a small-diameter endoscope, the contents of the spinal subarachnoid space could be examined. Further improvements to increase possible endoscopic manipulation and enhance safety may extend the possibilities for endoscopic examination and permit endoscopic treatment.


[Assessment of ischemic heart disease risk factors in relation to mortality (data from a multiyear prospective study in Moscow)].
Seven-year prospective study of mortality among the selected male population born in 1909--1918 showed that in some cases the region of high death risk is noted not only in the upper but also in the lower decyls and that of low risk in the center of the distribution of some of the ischemic heart disease (IHD) risk factors. Differentiation of the whole population into groups of individuals with IHD and those without IHD according to the findings of the initial examination enabled the authors to distinguish various tendencies of changes in the death risk depending on the arterial pressure levels and the lipid content. In prolongation of the period of observation of the population it is necessary to take into consideration the variability of the risk factors as well as the development of new cases with IHD in the period after the first examination.


Experimental Investigation of Additive Manufacturing of Continuous Carbon Fiber Composites with Multifunctional Electro-Tensile Properties.
Manufacturing processes for monofunctional and multifunctional materials vary depending on the design optimization. Multifunctional continuous carbon fiber composites provide great potential in achieving coupled structural and electrical properties for their applications in aircraft, unmanned aircraft systems, and spacecraft. Proper optimization of tensile and electrical properties offers benefits early in the design and continuous operational safety phases to obtain coupled multifunctional properties. In this paper, fused filament fabrication additive manufacturing (AM) technique was used to fabricate continuous carbon fiber solid laminated composites test coupons. The proposed new method characterizes the electrical conductivity's coupled effects on the tensile properties, including the failure loads and modes. This paper addresses a novel way of integrating electrical function into the composites that significantly reduce weight, potentially replacing the bulky electrical wires. Tensile and electrical conductivity tests were concurrently conducted on coupons, and the results were plotted and tabulated. The results showed the multifunctional properties of the maximum ultimate tensile strength of 392 MPa with the maximum tensile load of 8907 N, and resistance of 37.5 G·Ω. The average values for ultimate tensile strength and maximum load were 371 MPa and 8459 N, respectively.


C1q assay for the detection of complement fixing antibody to HLA antigens.
Solid phase Luminex(®) and flow cytometric single antigen bead assays offer exquisite sensitivity and specificity for HLA antibody detection. Unlike the historical complement-dependent cytotoxicity (CDC) method, these assays do not distinguish complement fixing from non-complement fixing antibody, the former of which are considered the most clinically relevant in the peri-transplant period. This chapter describes a novel solid phase C1q binding assay to distinguish HLA antibodies that can bind the first component of complement (C1q). These antibodies have the capacity to initiate the complement cascade irrespective of whether that actually occurs. The C1q assay detects many more complement fixing antibodies than are observed by the less sensitive and less specific CDC assay.


Validation studies on the 5-hydroxy-L-[beta-11C]-tryptophan/PET method for probing the decarboxylase step in serotonin synthesis.
The two-tissue compartment model, including irreversible trapping in the second compartment (2TCM) is used to describe the kinetics of 5-Hydroxy-L-[beta-(11)C]-tryptophan ([(11)C]HTP), a radioligand used in positron emission tomography (PET) for probing the second enzymatic step in the biosynthesis of serotonin. In this study, we examined the capacity of the model to track pharmacological changes in this biological process. We also investigated the potential loss of [(11)C]HTP-derived radioactivity during a PET study, since loss should be negligible not to alter quantification. Six rhesus monkeys were investigated using bolus [(11)C]HTP/PET methodology before and after pharmacological intervention. The second enzymatic step in serotonin synthesis was inhibited using the aromatic L-amino acid decarboxylase inhibitor NSD1015 (10 mg/kg). The extent of [(11)C]-derived radioactivity loss from the brain was studied by inhibition of the enzyme responsible for formation of the tissue metabolite, monoamine oxidase A, using clorgyline (2 mg/kg). After NSD1015, the uptake of [(11)C]HTP-derived radioactivity was increased in all the investigated brain regions, while the parameter used to reflect decarboxylase activity, the net accumulation rate constant (K(acc)), was decreased by 37% in the striatum, compared with baseline. Pretreatment with clorgyline did not change the brain uptake of [(11)C]HTP-derived radioactivity or K(acc). This study demonstrates that the 2TCM for [(11)C]HTP/PET is able to detect changes occurring during alteration of the biological process (i.e., the conversion of HTP to serotonin). Elimination of the radiotracer metabolite [(11)C]HIAA from the brain may be considered negligible if the PET study is limited to 60 min.


Selective Photoexcitation of Finite-Momentum Excitons in Monolayer MoS2 by Twisted Light.
Twisted light carries a well-defined orbital angular momentum (OAM) of lℏ per photon. The quantum number l of its OAM can be arbitrarily set, making it an excellent light source to realize high-dimensional quantum entanglement and ultrawide bandwidth optical communication structures. In spite of its interesting properties, twisted light interaction with solid state materials, particularly two-dimensional materials, is yet to be extensively studied via experiments. In this work, photoluminescence (PL) spectroscopy studies of monolayer molybdenum disulfide (MoS2), a material with ultrastrong light-matter interaction due to reduced dimensionality, are carried out under photoexcitation of twisted light. It is observed that the measured spectral peak energy increases for every increment of l of the incident light. The nonlinear l-dependence of the spectral blue shifts is well accounted for by the analysis and computational simulation of this work. More excitingly, the twisted light excitation revealed the unusual lightlike exciton band dispersion of valley excitons in monolayer transition metal dichalcogenides. This linear exciton band dispersion is predicted by previous theoretical studies and evidenced via this work's experimental setup.


Polymerisation shrinkage around composite resin restorations--an in vitro study.
The aim of this study was to determine the degree of penetration of a fluorescent adhesive at the margins of 3 brands of posterior composite resin restorative materials. Seventy-eight extracted human premolar teeth were used. Class II cavities were prepared on both approximal surfaces of each tooth in such a manner that the gingival wall was situated apical to the amelo-cemental junction. The enamel of the cavity walls was bevelled, etched and treated with the enamel bonding agent recommended by the manufacturer. The 3 brands of composite resin used were: Fulfil, Estilux Posterior and P 50. The teeth were divided at random into 3 groups of 26 premolars each. The mesial and distal cavities of each tooth in the group were filled with the same brand. The resin was inserted in two increments in both the mesial and distal cavities and polymerised. The margins of one restoration in each tooth was coated with fluorescent adhesive and cured. All the restored teeth were embedded in clear epoxy resin and then sectioned. The mounted sections were examined at 100X magnification with fluorescent light. The polymerisation contraction gap at different areas of the tooth/restoration interface and the degree of penetration of the fluorescent adhesive into the gap was determined. The fluorescent adhesive penetrated the contraction gaps in all 3 posterior restorative materials to varying degrees. The defects observed were similar in all 3 brands and the differences in degree of penetration were not statistically significant (p > 0.05).


Estimating health state utilities associated with a rare disease: familial chylomicronemia syndrome (FCS).
Aims: Familial chylomicronemia syndrome (FCS) is a rare genetic disorder with no currently approved therapies. Treatments are in development, and cost-utility analyses will be needed to examine their value. These models will require health state utilities representing FCS. Therefore, the purpose of this study was to estimate utilities for FCS and an associated episode of acute pancreatitis (AP).Methods: Because it is not feasible to gather a large enough sample of patients with this extremely rare condition to complete standardized preference-based measures, vignette-based methods were used to estimate utilities. In time trade-off interviews, general population participants in the UK and Canada valued health state vignettes drafted based on literature review, clinician input, and interviews with patients. Four health states described variations of FCS. A fifth health state, describing AP, was added to one of the other health states to evaluate its impact on utility.Results: A total of 308 participants provided utility data (208 UK; 100 Canada). Mean utilities for FCS health states ranged from 0.46 to 0.83, with higher triglycerides, more severe symptoms, and a history of AP associated with lower utility values. The disutility (i.e. utility decrease) of AP ranged from -0.17 to -0.25, with variations depending on the health state to which it was added. Utility means were similar in the UK and Canada.Conclusions: The vignette-based approach is useful for estimating utilities of a rare disease. The health state utilities derived in this study would be useful in models examining cost-effectiveness of treatments for FCS.


[Clinical response and survival in metastatic renal carcinoma during subcutaneous administration of interleukin-2 alone. Subcutaneous Il-2 in renal carcinoma].
Several clinical studies have demonstrated the efficacy of subcutaneous immunotherapy with Il-2 alone in metastatic renal cell carcinoma (RCC). In an attempt to better define the clinical parameters which may predict the efficacy of treatment, the present study shows the results obtained with subcutaneous Il-2 alone in 91 evaluable metastatic RCC patients. IL-2 was injected subcutaneously at 3 million IU twice/day for 5 days/week for 6 weeks, corresponding to one immunotherapeutic cycle. In nonprogressing patients, a second cycle was given after 28-day rest period. A complete response (CR) was achieved in 2/91 patients. Moreover, 19/91 patients had a partial response (PR). Therefore, objective response (OR) rate was 21/91 (23%) patients. Stable disease (SD) was achieved in 41 patients, while the remaining 29 patients had a progressive disease (PD). OR rate was significantly higher in patients with a long disease-free survival than in patients with synchronous metastases, in nephrectomized patients than in the non-nephrectomized ones, and in patients with high than in those with low PS. The survival obtained in patients with CR or PA was significantly longer with respect to that found in patients with SD or PD. The toxicity was substantially low in all patients. This study confirms that the subcutaneous immunotherapy with IL-2 alone is an effective and well tolerated therapy of metastatic RCC.


Involvement of aldosterone and mineralocorticoid receptors in rat mesangial cell proliferation and deformability.
We demonstrated recently that chronic administration of aldosterone to rats induces glomerular mesangial injury and activates mitogen-activated protein kinases including extracellular signal-regulated kinases 1/2 (ERK1/2). We also observed that the aldosterone-induced mesangial injury and ERK1/2 activation were prevented by treatment with a selective mineralocorticoid receptor (MR) antagonist, eplerenone, suggesting that the glomerular mesangium is a potential target for injuries induced by aldosterone via activation of MR. In the present study, we investigated whether MR is expressed in cultured rat mesangial cells (RMCs) and involved in aldosterone-induced RMC injury. MR expression and localization were evaluated by Western blotting analysis and fluorolabeling methods. Cell proliferation and micromechanical properties were determined by [3H]-thymidine uptake measurements and a nanoindentation technique using an atomic force microscope cantilever, respectively. ERK1/2 activity was measured by Western blotting analysis with an anti-phospho-ERK1/2 antibody. Protein expression and immunostaining revealed that MR was abundant in the cytoplasm of RMCs. Aldosterone (1 to 100 nmol/L) dose-dependently activated ERK1/2 in RMCs with a peak at 10 minutes. Pretreatment with eplerenone (10 micromol/L) significantly attenuated aldosterone-induced ERK1/2 phosphorylation. Aldosterone (100 nmol/L) treatment for 30 hours increased [3H]-thymidine incorporation and decreased the elastic modulus, indicating cellular proliferative and deforming effects of aldosterone, respectively. These aldosterone-induced changes in cellular characteristics were prevented by pretreatment with eplerenone or an ERK (MEK) inhibitor, PD988059 (100 micromol/L). The results indicate that aldosterone directly induces RMC proliferation and deformability through MR and ERK1/2 activation, which may contribute to the pathogenesis of glomerular mesangial injury.


Reliability and validity of the Chronic Liver Disease Questionnaire (CLDQ) in adults with non-alcoholic steatohepatitis (NASH).
Significant impairments in health-related quality of life (HRQL) in patients with non-alcoholic fatty liver disease have been previously described. The disease-specific HRQL among patients with non-alcoholic steatohepatitis (NASH), however, remains unknown. To determine the degree of construct validity of the Chronic Liver Disease Questionnaire (CLDQ) in adults with NASH. Participants referred for the evaluation of histology-proven NASH at Mayo Clinic, Rochester, between 1996 and 2000, were evaluated. HRQL assessment by the Short-Form 36 (SF-36) Health Survey and CLD) was performed. The primary outcome was to determine the level of correlation between overall and subscale scores for the CLDQ and SF-36 instruments. Among 79 participants (70%) with NASH completing both questionnaires (mean age, 51.2 years with 64% female gender), excellent reliability was noted for the CLDQ instrument. Significant reductions in all SF-36 domains (p<0.05 for all) including PCS and MCS scores (p<0.02 for both) among participants with NASH compared with normative data from an age-matched and sex-matched US general population sample was observed. Highly significant correlations were observed between overall CLDQ score with SF-36 PCS (r=0.82, p<0.0001) and SF-36 MCS (r=0.67, p<0.0001) scores. Similar degrees of correlation were observed between relevant subscales of the CLDQ and SF-36 as well. The CLDQ has excellent reliability and validity of construct for HRQL assessment in adults with NASH when compared with the SF-36. Future investigations among participants with NASH require assessing the responsiveness of the CLDQ to medical therapies and disease progression.


Concurrent X chromosome inactivation and upregulation during non-human primate preimplantation development revealed by single-cell RNA-sequencing.
In mammals, dosage compensation of X-linked gene expression between males and females is achieved by inactivation of a single X chromosome in females, while upregulation of the single active X in males and females leads to X:autosome dosage balance. Studies in human embryos revealed that random X chromosome inactivation starts at the preimplantation stage and is not complete by day 12 of development. Alternatively, others proposed that dosage compensation in human preimplantation embryos is achieved by dampening expression from the two X chromosomes in females. Here, we characterize X-linked dosage compensation in another primate, the marmoset (Callithrix jacchus). Analyzing scRNA-seq data from preimplantation embryos, we detected upregulation of XIST at the morula stage, where female embryos presented a significantly higher expression of XIST than males. Moreover, we show an increase of X-linked monoallelically expressed genes in female embryos between the morula and late blastocyst stages, indicative of XCI. Nevertheless, dosage compensation was not achieved by the late blastocyst stage. Finally, we show that X:autosome dosage compensation is achieved at the 8-cell stage, and demonstrate that X chromosome dampening in females does not take place in the marmoset. Our work contributes to the elucidation of primate X-linked dosage compensation.


Reorganization of columnar architecture in the growing visual cortex.
Many cortical areas increase in size considerably during postnatal development, progressively displacing neuronal cell bodies from each other. At present, little is known about how cortical growth affects the development of neuronal circuits. Here, in acute and chronic experiments, we study the layout of ocular dominance (OD) columns in cat primary visual cortex during a period of substantial postnatal growth. We find that despite a considerable size increase of primary visual cortext, the spacing between columns is largely preserved. In contrast, their spatial arrangement changes systematically over this period. Whereas in young animals columns are more band-like, layouts become more isotropic in mature animals. We propose a novel mechanism of growth-induced reorganization that is based on the "zigzag instability," a dynamical instability observed in several inanimate pattern forming systems. We argue that this mechanism is inherent to a wide class of models for the activity-dependent formation of OD columns. Analyzing one representative of this class, the Elastic Network model, we show that this mechanism can account for the preservation of column spacing and the specific mode of reorganization of OD columns that we observe. We conclude that column width is preserved by systematic reorganization of neuronal selectivities during cortical expansion and that this reorganization is well described by the zigzag instability. Our work suggests that cortical circuits may remain plastic for an extended period in development to facilitate the modification of neuronal circuits to adjust for cortical growth.


Preoperative Therapy for Unresectable Locally Advanced Pancreatic Cancer for ≥6 Months Improves Prognosis After Pancreatectomy.
This study aimed to identify the optimal duration of pretreatment for unresectable locally advanced (UR-LA) pancreatic cancer and analyze its effect on the prognosis. This retrospective study included 39 patients with UR-LA pancreatic cancer after pancreatectomy. The cutoff period of preoperative therapy was determined using a receiver operating characteristic curve. We investigated the relationship between preoperative and intraoperative clinical variables and overall survival (OS) in univariate and multivariate analyses. The relationship between the preoperative therapy duration and the clinicopathological variables was investigated. OS was compared according to preoperative therapy duration and the presence or absence of adjuvant surgery. After pretreatment, 15 patients underwent adjuvant surgery and 24 patients continued on chemotherapy without surgery. The multivariate analysis demonstrated preoperative therapy duration ≥6 months was an independent prognostic factor [hazard ratio (HR)=0.10, p=0.04]. No significant difference in the clinicopathological variables was observed between the two groups according to preoperative therapy duration. The OS was significantly better in patients who underwent adjuvant surgery after preoperative therapy duration ≥6 months than in those after preoperative therapy duration <6 months and in those without adjuvant surgery (5-year OS rates: 80% vs. 0%; p=0.01 and 5-year OS rates: 80% vs. 0%; p=0.004, respectively). The OS was not significantly better in patients with adjuvant surgery after preoperative therapy duration <6 months than in those without adjuvant surgery (2-year OS rates: 45.7% vs. 38.1%; p=0.98). Preoperative therapy for UR-LA pancreatic cancer for ≥6 months is necessary to improve prognosis after adjuvant surgery.


Escherichia coli alkaline phosphatase. An analysis of transient kinetics.
1. The hydrolysis of 2,4-dinitrophenyl phosphate by Escherichia coli alkaline phosphatase at pH5.5 was studied by the stopped-flow technique. The rate of production of 2,4-dinitrophenol was measured both in reactions with substrate in excess of enzyme and in single turnovers with excess of enzyme over substrate. It was found that the step that determined the rate of the transient phase of this reaction was an isomerization of the enzyme occurring before substrate binding. 2. No difference was observed between the reaction after mixing a pre-equilibrium mixture of alkaline phosphatase and inorganic phosphate, with 2,4-dinitrophenyl phosphate at pH5.5 in the stopped-flow apparatus, and the control reaction in which inorganic phosphate was pre-equilibrated with the substrate. Since dephosphorylation is the rate-limiting step of the complete turnover at pH5.5, this observation suggests that alkaline phosphatase can bind two different ligands simultaneously, one at each of the active sites on the dimeric enzyme, even though only one site is catalytically active at any given time. 3. Kinetic methods are outlined for the distinction between two pathways of substrate binding, which include an isomerization either of the free enzyme or of the enzyme-substrate complex.


Accommodation in renal transplantation: unanswered questions.
Accommodation, an acquired resistance of an organ to immune-mediated damage, has been recognized as an outcome of renal transplantation for more than 20 years. Accommodation was originally identified in blood group-incompatible kidney transplants that survived and functioned normally in recipients with high titers of antiblood group antibodies directed against antigens in the grafts. The most compelling questions today include how often and by which mechanisms accommodation occurs, and what might be the biological implications of accommodation. This communication summarizes recent advances in addressing these questions. Because its diagnosis has depended on identification of antidonor antibodies in serum, the prevalence of accommodation has been considered low. Recent research in animal models and clinical subjects may challenge that view. This research also suggests that sublethal graft injury of various types induces accommodation and that accommodation may be a dynamic condition, eventuating into tolerance on the one hand and chronic graft injury on the other. Burgeoning lines of investigation into accommodation now portray a condition of greater prevalence than once thought, exposing pathways that may contribute to the understanding of a range of responses to transplantation.


Hydronephrotic kidney with multiple extra-renal calyces.
Extra-renal calyces are a rare anomaly of the renal collecting system, characterised by the presence of calyces and pelvis outside the renal parenchyma. It may also be associated with other anomalies, such as renal ectopia, fusion and malrotation. We describe an unusual case of extra-renal calyces with six long calyces arising from a suspected multicystic dysplastic kidney that was successfully managed laparoscopically.


The nucleocapsid protein isolated from HIV-1 particles binds zinc and forms retroviral-type zinc fingers.
The role of zinc in retroviral gag protein function has been addressed through the application of high-resolution nuclear magnetic resonance spectroscopy to samples of the nucleocapsid protein (NCP, p7) isolated directly from infectious HIV-1 particles. Unlike reports for the NCP from avian myeloblastosis virus (AMV) particles [Jentoft et al. (1988) Proc. Natl. Acad. Sci. U.S.A. 85, 7094], we find that the HIV-1 NCP binds 2 equiv of zinc tightly and stoichiometrically. Two-dimensional NMR spectroscopic studies reveal that zinc binding induces formation of folded domains that are conformationally similar to (if not identical with) structures observed previously for relevant retroviral-type (RT) zinc finger peptides [formerly called zinc fingerlike peptides; Summers et al. (1990) Biochemistry 29, 329]. This finding is consistent with the hypothesis that the inability of mutant proteins (with substituted Cys and His residues) to package viral RNA results from deficient zinc-binding capability, which may have significant consequences in the development of vaccines for the prevention of AIDS.


Postnatal development of extracellular matrix and vascular function in small arteries of the rat.
Introduction: Vascular extracellular matrix (ECM) is dominated by elastic fibers (elastin with fibrillin-rich microfibrils) and collagens. Current understanding of ECM protein development largely comes from studies of conduit vessels (e.g., aorta) while resistance vessel data are sparse. With an emphasis on elastin, we examined whether changes in postnatal expression of arteriolar wall ECM would correlate with development of local vasoregulatory mechanisms such as the myogenic response and endothelium-dependent dilation. Methods: Rat cerebral and mesenteric arteries were isolated at ages 3, 7, 11, 14, 19 days, 2 months, and 2 years. Using qPCR mRNA expression patterns were examined for elastin, collagen types I, II, III, IV, fibrillin-1, and -2, lysyl oxidase (LOX), and transglutaminase 2. Results: Elastin, LOX and fibrillar collagens I and III mRNA peaked at day 11-14 in both vasculatures before declining at later time-points. 3D confocal imaging for elastin showed continuous remodeling in the adventitia and the internal elastic lamina for both cerebral and mesenteric vessels. Myogenic responsiveness in cannulated cerebral arteries was detectable at day 3 with constriction shifted to higher intraluminal pressures by day 19. Myogenic responsiveness of mesenteric vessels appeared fully developed by day 3. Functional studies were performed to investigate developmental changes in endothelial-dependent dilation. Endothelial-dependent dilation to acetylcholine was less at day 3 compared to day 19 and at day 3 lacked an endothelial-derived hyperpolarizing factor component that was evident at day 19. Conclusion: Collectively, in the rat small artery structural remodeling and aspects of functional control continue to develop in the immediate postnatal period.


Long-term efficacy and toxicity of abacavir/lamivudine/nevirapine compared to the most prescribed ARV regimens before 2013 in a French Nationwide Cohort Study.
Data on the long-term efficacy and safety of abacavir/lamivudine (ABC/3TC) and nevirapine (NVP) are scarce. This combination has the advantage of simplifying treatment and improving long-term tolerance. The aim of this study was to compare the rate of any discontinuation of antiretroviral (ARV) regimen because of virologic failure (VF), and/or adverse drug reaction (ADR) among patients receiving stable ARV regimens for at least 6 months.ABC/3TC/NVP was compared to ABC/3TC with either ritonavir-boosted darunavir (DRV/r) or ritonavir-boosted atazanavir (ATV/r), unboosted ATV, or tenofovir/emtricitabine (TDF/FTC) with either one of the following: ATV/r, unboosted ATV, DRV/r, efavirenz (EFV), or NVP, in the French prospective multicenter Dat'AIDS cohort.The study enrolled 16,511 patients treated with following ARV regimens: ABC/3TC/NVP (n = 1089), TDF/FTC/NVP (n = 1542), ABC/3TC/DRV/r (n = 1065), ABC/3TC/ATV/r (n = 1847), ABC/3TC/ATV (n = 563), TDF/FTC/ATV/r (n = 3519), TDF/FTC/DRV/r (n = 2767), TDF/FTC/ATV (n = 419), and TDF/FTC/EFV (n = 3700). Mean follow-up was 36 ± 24 months. Patients treated with ABC/3TC/NVP received this regimen as a switch regimen in 97% of cases. By multivariable analysis, the risk of treatment discontinuation due to VF was similar between ABC/3TC/NVP and other ARV regimens, except for TDF/FTC/ATV and ABC/3TC/ATV, which were associated with a higher risk of treatment interruption due to VF (hazard ratio [HR] 1.99; 95% confidence interval [CI] 1.29-3.06 and HR 2.19; 95% CI 1.51-3.18, respectively). Treatment discontinuation due to ADR was lowest with the ABC/3TC/NVP regimen. Other ARV regimens were associated with a 1.80- to 3.19-fold increase in the risk of treatment discontinuation due to ADR (P < 0.0001 for all comparisons).ABC/3TC/NVP as a simplification regimen is a long-term effective regimen with lower discontinuation due to long-term toxicity compared with other standard ARV regimens.


Serum Bile Acids in Patients With Primary Open-angle Glaucoma.
To measure the serum bile acids (SBA) in patients with primary open-angle glaucoma (POAG) and to compare with nonglaucomatous controls. A hospital-based case control study of 90 patients above the age of 40 years was conducted. Patients with POAG formed the study group and individuals without glaucoma in the same age group formed the control group. SBA estimation was done using ELISA kit enzymatic assay technique and the median value was compared between the 2 groups. The median of SBA level was found to be more in POAG patients when compared with the control group. The difference was statistically significant (P<0.001). Median SBA level was not affected by the age or sex of the participants in the study. "Bile acids" may have a role in the oxidative stress and apoptosis involved in the pathophysiology of POAG.


The role of cardiac biomarkers for predicting left ventricular dysfunction and cardiovascular mortality in acute exacerbations of COPD.
The presence of cardiovascular comorbidities is frequently associated with poor outcomes in chronic obstructive pulmonary disease (COPD). No clear role has been defined for cardiac biomarkers in acute exacerbations of COPD (AECOPD). The aim of this systematic review was to examine the prognostic value of brain natriuretic peptide (BNP) and troponins in patients with AECOPD. Two independent authors searched the PubMed and Cochrane Library to collect clinical trials, observational studies and meta-analyses studying the prognostic value of cardiac biomarkers in AECOPD. The reference lists of all the included studies were also reviewed. A total of 14 studies were included in the review, of which 10 measured troponins, 7 measured BNP or NT-proBNP, and 3 measured both. Of the studies that used mortality in AECOPD as an end point, some but not all found that elevated BNP and/or troponins were associated with increased mortality. Of the studies that used left ventricular (LV) dysfunction in AECOPD as an end point, all found a significant association between elevated BNP and troponins in the diagnosis of LV dysfunction. In summary, it appears that there may be a link between an elevated level of BNP or NT-proBNP and increased cardiovascular mortality in AECOPD, although the data currently available are not conclusive. The inconsistencies in biomarkers measured, time points of measurements and the variability in outcome measured preclude more robust analysis.


Roller Massage Prior to Running Does Not Affect Gait Mechanics in Well-Trained Runners.
Understanding if roller massage prior to a run can mitigate fatigue-related decrements in muscle force production during prolonged running is important because of the association between fatigue and running-related injury. The authors investigated whether a bout of roller massage prior to running would (1) mitigate fatigue-related increases in vertical average load rate and free moment of the ground reaction force of running and (2) mitigate decreases in maximal countermovement jump height. Repeated-measures study. Laboratory. A total of 14 recreational endurance athletes (11 men and 3 women) volunteered for the study. A 12.5-minute foam roller protocol for the lower extremities and a fatiguing 30-minute treadmill run. Vertical average load rate, free moment, and maximal jump height before (PRE) and after (POST) the fatiguing treadmill run on separate experimental days: once where participants sat quietly prior to the fatiguing run (REST) and another where the foam roller protocol was performed prior to the run (ROLL). A 2-way multiple analysis of variance found no significant differences in vertical average load rate, free moment, and jump height between PRE/POST times in both REST/ROLL conditions. The authors concluded that recreational endurance athletes maintain running mechanics and jump performance after a fatiguing run regardless of prerun roller massage and may not rely on prerun roller massage as a form of injury prevention.


Phytochemical profiles, antioxidant and antimicrobial activities of three Potentilla species.
Extracts from Potentilla species have been applied in traditional medicine and exhibit antioxidant, hypoglycemic, anti-inflammatory, antitumor and anti-ulcerogenic properties, but little has been known about the diversity of phytochemistry and pharmacology on this genus. This study investigated and compared the phytochemical profiles, antioxidant and antimicrobial activities of leaf extracts from three Potentilla species (Potentilla fruticosa, Potentilla glabra and Potentilla parvifolia) in order to discover new resources for lead structures and pharmaceutical products. Chemical composition and content of six phenolic compounds were evaluated and determined by RP-HPLC; Total phenolic and total flavonoid content were determined using Folin-Ciocalteau colourimetric method and sodium borohydride/chloranil-based method (SBC); Antioxidant activities were determined using DPPH, ABTS and FRAP assays; Antimicrobial properties were investigated by agar dilution and mycelial growth rate method. The results showed hyperoside was the predominant phenolic compound in three Potentilla species by RP-HPLC assay, with the content of 8.86 (P. fruticosa), 2.56 (P. glabra) and 2.68 mg/g (P. parvifolia), respectively. The highest content of total identified phenolic compounds (hyperoside, (+)-catechin, caffeic acid, ferulic acid, rutin and ellagic acid) was observed in P. parvifolia (14.17 mg/g), follow by P. fruticosa (10.01 mg/g) and P. glabra (7.01 mg/g). P. fruticosa possessed the highest content of total phenolic (84.93 ± 0.50 mmol gallic acid equivalent/100 g) and total flavonoid (84.14 ± 0.03 mmol quercetin equivalent/100 g), which were in good correlation with its significant DPPHIC50 (16.87 μg/mL), ABTS (2763.48 μmol Trolox equivalent/g) and FRAP (1398.70 μmol Trolox equivalent/g) capacities. Furthermore, the effective methodology to distinguish the different species of Potentilla was also established by chromatographic fingerprint analysis for the first time. The results of antimicrobial activities showed P. fruticosa exhibited the strongest inhibition aganist Gram-positive bacteria, Pseudomonas aeruginosa and Candida albicans with MIC values of 0.78-6.25 mg/mL. P. parvifolia possessed antibacterial and antifungal activities against all the microorganisms tested, with EC50 and MIC values of 20.52-47.02 mg/mL and 0.78-50 mg/mL, respectively. These results indicated that leaf extracts from three Potentilla species could become useful supplement for pharmaceutical products as a new antioxidant and antimicrobial agents.


Performance enhancement of a graphene-zinc phosphide solar cell using the electric field-effect.
The optical transparency and high electron mobility of graphene make it an attractive material for photovoltaics. We present a field-effect solar cell using graphene to form a tunable junction barrier with an Earth-abundant and low cost zinc phosphide (Zn3P2) thin-film light absorber. Adding a semitransparent top electrostatic gate allows for tuning of the graphene Fermi level and hence the energy barrier at the graphene-Zn3P2 junction, going from an ohmic contact at negative gate voltages to a rectifying barrier at positive gate voltages. We perform current and capacitance measurements at different gate voltages in order to demonstrate the control of the energy barrier and depletion width in the zinc phosphide. Our photovoltaic measurements show that the efficiency conversion is increased 2-fold when we increase the gate voltage and the junction barrier to maximize the photovoltaic response. At an optimal gate voltage of +2 V, we obtain an open-circuit voltage of V oc = 0.53 V and an efficiency of 1.9% under AM 1.5 1-sun solar illumination. This work demonstrates that the field effect can be used to modulate and optimize the response of photovoltaic devices incorporating graphene.


Defatted flaxseed flour improves weight loss and lipid profile in overweight and obese adults: a randomized controlled trial.
Studies have suggested that foods rich in dietary fiber may contribute to body weight loss and lower triacylglycerol (TG) levels. This study aimed to investigate the effect of flaxseed meal (FM) (a by-product of flaxseed after oil processing) supplemented biscuits on body weight, body composition, and blood lipids in overweight and obese participants. In a double-blind randomized controlled trial, 53 overweight and obese adults (18-36 years of age) were recruited and randomized to consume control biscuits or biscuits supplemented with FM for their breakfast for 60 days (approximately 100 g per day). Significant group × time interaction (P = 0.011) was observed for body weight, and the body weight of the FM group was significantly lower than that of the control group (P = 0.049). We also found significant group × time interactions for body mass index (BMI) (P = 0.008), TG (P = 0.041) and interleukin-6 (IL-6) (P = 0.016). In addition, after 60 days of intervention, the body weight, BMI and TG levels of the FM group significantly decreased compared to those of the control group. On day 60, the serum concentration of IL-6 in the FM group was significantly lower than that in the control group. FM supplemented biscuits have a beneficial effect on body weight, BMI and TG of overweight and obese subjects (ClinicalTrials.gov registration number: ChiCTR1900022833).


Continuous estimation of finger joint angles using muscle activation inputs from surface EMG signals.
Prediction of dynamic hand finger movements has many clinical and engineering applications in the control of human interface devices such as those used in virtual reality control, robot prosthesis and rehabilitation aids. Surface electromyography (sEMG) signals have often been used in the mentioned applications because these reflect the motor intention of users very well. In this study, we present a method to estimate the finger joint angles of a hand from sEMG signals that considers electromechanical delay (EMD), which is inherent when EMG signals are captured alongside motion data. We use the muscle activation obtained from the sEMG signals as input to a neural network. In this muscle activation model, the EMD is parameterized and automatically obtained through optimization. With this method, we can predict the finger joint angles with sEMG signals in both periodic and nonperiodic free movements of the flexion and extension movement of the fingers. Our results show correlation as high as 0.92 between the actual and predicted metacarpophalangeal (MCP) joint angles for periodic finger flexion movements, and as high as 0.85 for nonperiodic movements, which are more dynamic and natural.


Fruit and vegetable intake and risk of amyotrophic lateral sclerosis in Japan.
There has been little interest in the role of nutrition in the prevention of amyotrophic lateral sclerosis (ALS). We investigated the relationship between dietary intake of vegetables, fruit, and antioxidants and the risk of ALS in Japan. Between 2000 and 2004, we recruited 153 ALS patients aged 18-81 years with disease duration of 3 years within the study period in accordance with El Escorial World Federation of Neurology criteria. Three hundred and six gender- and age-matched controls were randomly selected from the general population. Information on dietary factors was collected using a validated self-administered diet history questionnaire. A higher consumption of all fruits and vegetables and fruit alone in the highest quartiles was associated with a statistically significantly reduced risk of ALS. Although not statistically significant, a beneficial association between intake of all vegetables, green and yellow vegetables and other vegetables and ALS was found. No statistically significant dose-response relationship was observed between intake of beta-carotene, vitamin C and vitamin E and the risk of ALS. Our findings suggest that higher intake of food rich in antioxidants such as fruit and vegetables confer protection against the development of ALS.


Interaction effects of oxytocin receptor gene polymorphism and depression on hippocampal volume.
Many studies have revealed that the oxytocin receptor gene (OXTR) is associated with emotional salience and depression among females. Hippocampus is closely associated with the pathophysiology of major depressive disorder (MDD). However, little is known of the interaction effects of OXTR and MDD on hippocampal volume. We sought to investigate the interaction effects of OXTR (rs53576) allelic variants and MDD on hippocampal volumes which also including subfield volumes. The OXTR rs53576 genotype groups were categorized as minor G allele carriers and A allele homozygotes. A total of 47 female patients with depression and 30 healthy females were included in this study. There were significant interactions between OXTR allele type and diagnosis of MDD on the 7 hippocampal subfield volumes, such as left presubiculum, left subiculum, left molecular, right cornus ammonis 1, right granule cells in the molecular layer of the dentate gyrus, right molecular layer, and right subiculum. There were no differences in the hippocampal volumes between MDD vs healthy controls or OXTR A vs G alleles. Our results demonstrate the importance of the interactions between OXTR and MDD on hippocampal volume. Future studies with large sample size should expand those interactions in the whole brain volumes.


A method using 4D dose accumulation to quantify the interplay effect in lung stereotactic body radiation therapy.
The purpose of this study was to devise and evaluate a method to quantify the dosimetric uncertainty produced by the interplay between the movement of multileaf collimator and respiratory motion in lung stereotactic body radiation therapy. The method calculates the dose distribution for all control points from a dynamic treatment in all respiratory phases. The methodology includes some characteristics of a patient's irregular breathing patterns. It selects, for each control point, the phases with maximum and minimum mean dose over the tumor and their corresponding adjacent phases, whenever necessary. According to this selection, the dose matrices from each control point are summed up to obtain two dose distributions in each phase, which are accumulated in the reference phase subsequently by deformable image registration (DIR). D 95 and [Formula: see text] were calculated over those accumulated dose distributions for Gross Tumor Volume (GTV), Planning Target Volume-based on Internal Target Volume approach-and Evaluation Target Volume (ETV), a novel concept that applies to 4D dose accumulation. With the ETV, DIR and interplay uncertainties are separated. The methodology also evaluated how variations in the breathing rate and field size affects the mean dose received by the GTV. The method was applied retrospectively in five patients treated with intensity modulated radiotherapy-minimum area defined by the leaves configuration at any control point was at least 4 cm2. Uncertainties in tumor coverage were small (in most patients, changes on D 95 and [Formula: see text] were below 2% for GTV and ETV) but significant over- and under-dosages near ETV, which can be accentuated by highly irregular breathing. Uncertainties in mean dose for GTV tended to decrease exponentially with increasing field size and were reduced by an increase of breathing rate. The implementation of this method would be helpful to assess treatment quality in patients with irregular breathing. Furthermore, it could be used to study interplay uncertainties when small field sizes are used.


[Healthcare for teenagers: are we working together?].
There are about 1.8 million children between 10 and 18 years of age in the Netherlands in 2016. These teenagers account for approximately 10% of the total population. Teenagers are relatively healthy and do not make much use of curative care. However, they are an important group in terms of public health, because a basis for good health in later life is created in the teenage years. Good health in teenagers is also important for education, relationships and employment, and their health has an influence on the health of the next generation. Child and adolescent healthcare plays an important part in preventive care for teenagers. Better cooperation and exchange of information between paediatricians, specialists in child and adolescent healthcare and general practitioners are important in order to optimise care for teenagers.


Proteomic analysis of the oil palm fruit mesocarp reveals elevated oxidative phosphorylation activity is critical for increased storage oil production.
Palm oil is a highly versatile commodity with wide applications in the food, cosmetics, and biofuel industries. Storage oil in the oil palm mesocarp can make up a remarkable 80% of its dry mass, making it the oil crop with the richest oil content in the world. As such, there has been an ongoing interest in understanding the mechanism of oil production in oil palm fruits. To identify the proteome changes during oil palm fruit maturation and factors affecting oil yield in oil palm fruits, we examined the proteomic profiles of oil palm mesocarps at four developing stages--12, 16, 18, and 22 weeks after pollination--by 8-plex iTRAQ labeling coupled to 2D-LC and MALDI-TOF/TOF MS. It was found that proteins from several important metabolic processes, including starch and sucrose metabolism, glycolysis, pentose phosphate shunt, fatty acid biosynthesis, and oxidative phosphorylation, were differentially expressed in a concerted manner. These increases led to an increase in carbon flux and a diversion of resources such as ATP and NADH that are required for lipid biosynthesis. The temporal proteome profiles between the high-oil-yielding (HY) and low-oil-yielding (LY) fruits also showed significant differences in the levels of proteins involved in the regulation of the TCA cycle and oxidative phosphorylation. In particular, the expression level of the β subunit of the ATP synthase complex (complex IV of the electron transport chain) was found to be increased during fruit maturation in HY but decreased in the LY during the fruit maturation. These results suggested that increased energy supply is necessary for augmented oil yield in the HY oil palm trees.


SORCS1 contributes to the development of renal disease in rats and humans.
Many lines of evidence demonstrate that genetic variability contributes to chronic kidney disease susceptibility in humans as well as rodent models. Little progress has been made in discovering causal kidney disease genes in humans mainly due to genetic complexity. Here, we use a minimal congenic mapping strategy in the FHH (fawn hooded hypertensive) rat to identify Sorcs1 as a novel renal disease candidate gene. We investigated the hypothesis that genetic variation in Sorcs1 influences renal disease susceptibility in both rat and human. Sorcs1 is expressed in the kidney, and knocking out this gene in a rat strain with a sensitized genome background produced increased proteinuria. In vitro knockdown of Sorcs1 in proximal tubule cells impaired protein trafficking, suggesting a mechanism for the observed proteinuria in the FHH rat. Since Sorcs1 influences renal function in the rat, we went on to test this gene in humans. We identified associations between single nucleotide polymorphisms in SORCS1 and renal function in large cohorts of European and African ancestry. The experimental data from the rat combined with association results from different ethnic groups indicates a role for SORCS1 in maintaining proper renal function.


Variation of lipids and lipoproteins in premenopausal women compared with men and postmenopausal women. DELTA (Dietary Effects on Lipoproteins and Thrombogenic Activity) Investigators.
Numerous studies have reported cyclic fluctuations in lipids and lipoproteins as a function of the phase of the menstrual cycle. However, the reported patterns are quite variable and have led to an unclear picture of the influence of the menstrual cycle on the variability of lipids, and hence of the role of the menstrual cycle phase in the interpretation of serum lipids for premenopausal women. As part of the DELTA Study (Dietary Effects on Lipoproteins and Thrombogenic Activity), we evaluated the cyclic variation of circulating lipids and lipoproteins in 39 premenopausal women and compared intraindividual variances in these women, 18 postmenopausal women, and 46 men under conditions of tight dietary control. Cholesterol, high-density lipoprotein (HDL)-cholesterol, low-density lipoprotein (LDL)-cholesterol, triglyceride, apolipoproteins A-1 (apo A-1) and B-100 (apo B-100), and lipoprotein (a) [Lp(a)] all demonstrated cycling in the premenopausal women. However, the observed cycling accounts for only a small fraction of the total biologic variability of lipids in premenopausal women. The magnitude of total intraindividual variability based on coefficient of variation (CV) for these lipids in premenopausal women (CV, 4% to 8.1%) was similar to that found for men (CV, 4.3% to 9.1%) and for postmenopausal women (CV, 3.7% to 6.7%). These results suggest that protocols for screening and monitoring of serum lipids in premenopausal women need not differ from those used for men or postmenopausal women.


Nuclear Translocation of Argonaute 2 in Cytokine-Induced Senescence.
Cellular senescence, or permanent growth arrest, is known as an effective tumor suppressor mechanism that can be induced by different stressors, such as oncogenes, chemotherapeutics or cytokine cocktails. Previous studies demonstrated that the growth-repressing state of oncogene-induced senescent cells depends on argonaute protein 2 (Ago2)-mediated transcriptional gene silencing and Ago2/Rb corepression of E2F-dependent cell cycle genes. Cytokine-induced senescence (CIS) likewise depends on activation of the p16Ink4a/Rb pathway, and consecutive inactivation of the E2F family of transcription factors. In the present study, we therefore analyzed the role of Ago2 in CIS. Human cancer cell lines were treated with interferon-gamma (IFN-γ) and tumor necrosis factor (TNF) to induce senescence. Senescence was determined by growth assays and measurement of senescence-associated β-galactosidase (SA-β-gal) activity, Ago2 translocation by Ago2/ Ki67 immunofluorescence staining and western blot analysis, and gene transcription by quantitative polymerase chain reaction (qPCR). IFN-γ and TNF permanently stopped cell proliferation and time-dependently increased SA-β-gal activity. After 24 - 48 h of cytokine treatment, Ago2 translocated from the cytoplasm into the nucleus of Ki67-negative cells, an effect which was shown to be reversible. Importantly, the proinflammatory cytokine cocktail suppressed Ago2-regulated cell cycle control genes, and siRNA-mediated depletion of Ago2 interfered with cytokine-induced growth inhibition. IFN-γ and TNF induce a stable cell cycle arrest of cancer cells that is accompanied by a fast nuclear Ago2 translocation and repression of Ago2-regulated cell cycle control genes. As Ago2 downregulation impairs cytokine-induced growth regulation, Ago2 may contribute to tissue homeostasis in human cancers.