Oscillatory corticothalamic response to somatosensory input.
We construct a model of the thalamocortical feedback loop using the Random Neural Network model to provide a theoretical explanation for cortical and thalamic damped rapid (circa 60-100 Hz) oscillations which have been observed in lightly anesthesized rats with somatosensory input Nicolelis (1997). Experimental data corroborates the choice of model parameter settings. Model predictions indicate that positive feedback loops and large signaling delays within cortex significantly determine the oscillatory phenomenon and its duration (i.e. its decay), while positive feedback from cortex to thalamus is needed to exhibit oscillations in thalamus.


Use of Productivity Loss/Gain in Cost-Effectiveness Analyses for Drugs: A Systematic Review.
Inclusion of productivity losses and gains in cost-effectiveness analyses for drugs is recommended by pharmacoeconomic guidelines in some countries and is considered optional in others. Often guidelines recommend analysis based on the payer perspective, but suggest that a supplemental analysis based on the societal perspective may be submitted that includes productivity losses/gains. However, there is no universally recognized framework for the approach to including productivity losses and gains in pharmacoeconomic analyses. This study aimed to systematically review literature on cost-effectiveness analyses of drug interventions that included costs associated with productivity losses/gains and to summarize the types cost elements included and cost calculation methods employed. Moreover, this study examines variations in the calculation of productivity losses/gains by target disease type, geographic region, income group, period of analysis, and analysis time horizon-as well as the impact of their inclusion on the study outcomes. A search of three databases was performed, including MEDLINE, Embase, and the Cochrane Library, to identify cost-effectiveness and cost-utility analyses that included indirect costs such as productivity losses/gains. Publications from January 2010 to October 2019 were examined and selected for inclusion by two independent reviewers. In addition to the citation details, data on the country of analysis, country income group, target disease area, study sponsorship, type of analysis, study design, time horizon, analysis perspective, productivity loss/gain elements included, the approach used to estimate productivity losses/gains, and the impact of their inclusion on the study outcome-namely the incremental cost effectiveness ratio-were extracted and summarized. The search strategy identified 5038 unique studies, and 208 were included in the final analysis. Among the studies reviewed, 165 (79%) were conducted in high-income countries and 160 (77%) were conducted for North American and European/Central Asian countries. The productivity loss/gain elements included in the analysis were reported for 169 studies (81%). Absenteeism only was included for 98 studies (47%), and absenteeism plus presenteeism was included for 29 studies (14%). Absenteeism plus some other element such as costs associated with unemployment and/or early retirement was included for 32 studies (15%) examined. Only one out of four of the studies reviewed included information on the approach used to estimate productivity losses/gains, which was predominantly the human capital approach. One-hundred forty-four studies (69%) reported the impact of including productivity losses/gains on the ICER, with 110 studies (53%) reporting that their inclusion contributed to more favorable cost-effectiveness. Although inclusion of productivity losses/gains was shown to have a favorable impact on evaluations for many studies, their impact and method of calculation was often not reported or was unclear. Further examination and discussion is needed to consider the optimal framework for considering productivity losses/gains in cost-effectiveness analyses, including the appropriate cost elements to include (e.g., patient absenteeism, caregiver absenteeism, presenteeism, unemployment, etc.) and how those costs should be estimated.


Relationship between hospital volume and operative mortality for liver resection: Data from the Japanese Diagnosis Procedure Combination database.
The present study aimed to conduct a nationwide investigation on the relationship between hospital volume and outcomes following liver resection in Japan. We also discuss health policy implications of the results. Using the Japanese Diagnosis Procedure Combination database, we identified 18 046 patients who underwent hepatic resection between July and December 2007-2009. Patients were subdivided into hospital-volume quartiles: very low- (<18/year), low- (18-35), high- (36-70) and very high-volume groups (>70). Multivariate logistic regression analysis for in-hospital mortality within 30 days of surgery was performed to analyze adjusted effects of various factors. Patients in the very high-volume group had a higher Charlson Comorbidity Index (P < 0.001) than those in the very low-volume group. Very low-volume hospitals were significantly less likely to perform extended lobectomy than very high-volume hospitals (5.4% vs 17.6%, P < 0.001). Crude in-hospital mortality within 30 days of surgery was 1.1% (0.6%, 0.8%, 1.9% and 3.0% for limited resection, segmentectomy, lobectomy and extended lobectomy, respectively). With reference to the very low-volume group, risk-adjusted odds ratios (95% confidence intervals) of low-, high- and very high-volume groups for overall mortality were 0.70 (0.48-1.02; P = 0.060), 0.52 (0.34-0.81; P = 0.004) and 0.16 (0.09-0.30; P < 0.001), respectively. There is a linear trend between higher hospital volume and lower in-hospital mortality of liver resection in Japan, particularly for lobectomy and extended lobectomy. Based on these results, regionalization of lobectomy and extended lobectomy in high-volume centers could be effective for reducing postoperative mortality.


Optimization of date syrup for enhancement of the production of citric acid using immobilized cells of Aspergillus niger.
Date syrup as an economical source of carbohydrates and immobilized Aspergillus niger J4, which was entrapped in calcium alginate pellets, were employed for enhancing the production of citric acid. Maximum production was achieved by pre-treating date syrup with 1.5% tricalcium phosphate to remove heavy metals. The production of citric acid using a pretreated medium was 38.87% higher than an untreated one that consumed sugar. The appropriate presence of nitrogen, phosphate and magnesium appeared to be important in order for citric acid to accumulate. The production of citric acid and the consumed sugar was higher when using 0.1% ammonium nitrate as the best source of nitrogen. The production of citric acid increased significantly when 0.1 g/l of KH2PO4 was added to the medium of date syrup. The addition of magnesium sulfate at the rate of 0.20 g/l had a stimulating effect on the production of citric acid. Maximum production of citric acid was obtained when calcium chloride was absent. One of the most important benefits of immobilized cells is their ability and stability to produce citric acid under a repeated batch culture. Over four repeated batches, the production of citric acid production was maintained for 24 days when each cycle continued for 144 h. The results obtained in the repeated batch cultivation using date syrup confirmed that date syrup could be used as a medium for the industrial production of citric acid.


Ontogeny of swimming behavior and brain catecholamine turnover in rats prenatally exposed to a mixture of 2,4-dichlorophenoxyacetic and 2,4,5-trichlorophenoxyacetic acids.
Rats exposed in utero on gestational days 6-15, to nonfetotoxic and grossly nonteratogenic mixtures (50 or 100 mg/kg) of 2,4-D/2,4,5-T as found in Agent Orange (but without significant contamination with 2,3,7,8 tetrachloro-p-dioxin) manifested subtle developmental neurotoxicity. Maturation of swimming behavior was significantly delayed on postnatal day 7 in both treatment groups. The concentration of norepinephrine (NE) in whole brain was significantly increased on postnatal day 15 in both treatment groups, whereas the concentration of dopamine (DA) was increased on postnatal day 15 at 100 mg/kg. The turnover and efflux rate constant of DA in whole brain were significantly reduced whereas the turnover time increased on postnatal day 3. The efflux rate constant for NE decreased and the turnover time increased significantly on postnatal day 15 at 100 mg/kg. These data indicate the value of ontogenic assessment following exposure to small doses, which result in functional alterations in the absence of overt toxic signs.


Performance of rapid diagnostic tests for the detection of anti-HBs in various patient populations.
Rapid diagnostic tests (RDTs) represent an attractive alternative method to conventional diagnosis of hepatitis B virus (HBV) infection. The aim of the present study was to evaluate the diagnostic performance of commercially available RDTs for the detection of anti-HBs in various patient populations. A total of 347 individuals, 198 positive and 149 negative for anti-HBs, were studied. The specificity of RDT detection of anti-HBs in serum was 98.0%, 96.0% and 97.3% with TOYO® HBsAb Test, QuickProfile™ HBsAb test and QuickProfile™ HBV-3 Panel test, respectively. The diagnostic sensitivity varied between 60.4% and 69.5%. The sensitivity of the three RDTs was markedly better when testing serum samples with an anti-HBs titer higher than 100IU/L, and reached 90% or more for an anti-HBs titer above 150IU/L. This performance was disappointing because the assays were not sensitive enough to detect low antibody titers. Thus, these tests require further improvement before they can be widely used in clinical practice.


In silico designed microtubule-stabilizer drugs against tauopathy in Alzheimer's disease.
Microtubules are the main building blocks of the cytoskeleton that maintain the shape of the cell. Microtubule-associated proteins, such as Tau protein, facilitate their plasticity in cells. Highly phosphorylated Tau has weak affinity to microtubule and, hence, high probability of aggregation into neurofibrillary tangles (tauopathy). Alzheimer's disease evolves when Tau proteins are abnormally phosphorylated. To prevent tauopathy in Alzheimer's disease, we designed drugs de novo targeting them in silico to the phosphorylated Tau-microtubule complexes. Our molecular docking (AutoDock, MOE, GOLD) and molecular dynamics (GROMACS, 2019.6) simulation results revealed compound 23 (C12H28N4O5) as a potential drug candidate, since it can bind (-11.1 kcal/mol by AutoDock) and fix not only phosphorylated Tau on the surface of microtubules, but also prevent their aggregation into bundles. In addition, compound 23 has shown its ability to de-bundle already grouped phosphorylated peptides into single pieces.Communicated by Ramaswamy H. Sarma.


Pharmacological management of pain after periodontal surgery: a systematic review with meta-analysis.
To assess and compare the pharmacological effect of different drugs on pain relief after periodontal surgery. Five databases were searched up to September 2019. The eligible studies comprised randomized clinical trials, involving only adult individuals that received any periodontal surgery and presenting two distinct groups of therapeutic regimens to control postoperative pain. Placebo groups could be included. The risk of bias was assessed with the RoB 2 Cochrane tool and the GRADE system. Meta-analyses were performed using different follow-up and drug comparisons. Overall, 2398 studies were identified, of which 35 were included. Low risk of bias was determined for the majority of the studies. The meta-analyses showed that the comparison of dexamethasone or non-steroidal anti-inflammatory drugs (NSAID) versus placebo favored the use of both interventions in a follow-up of 1 to 8 h for open flap procedures (OFP). However, no statistical difference was found for the comparison between NSAID and dexamethasone for OFP. Patients may benefit from several pharmacological schemes for pain relief after periodontal surgeries. However, due to the high heterogeneity among studies, no fixed pharmacological protocol could be proposed. There is not enough evidence to recommend one therapeutic scheme. However, untreated pain is harmful to the patients and it is not advisable.


AMPK regulation of Raptor and TSC2 mediate metformin effects on transcriptional control of anabolism and inflammation.
Despite being the frontline therapy for type 2 diabetes, the mechanisms of action of the biguanide drug metformin are still being discovered. In particular, the detailed molecular interplays between the AMPK and the mTORC1 pathway in the hepatic benefits of metformin are still ill defined. Metformin-dependent activation of AMPK classically inhibits mTORC1 via TSC/RHEB, but several lines of evidence suggest additional mechanisms at play in metformin inhibition of mTORC1. Here we investigated the role of direct AMPK-mediated serine phosphorylation of RAPTOR in a new RaptorAA mouse model, in which AMPK phospho-serine sites Ser722 and Ser792 of RAPTOR were mutated to alanine. Metformin treatment of primary hepatocytes and intact murine liver requires AMPK regulation of both RAPTOR and TSC2 to fully inhibit mTORC1, and this regulation is critical for both the translational and transcriptional response to metformin. Transcriptionally, AMPK and mTORC1 were both important for regulation of anabolic metabolism and inflammatory programs triggered by metformin treatment. The hepatic transcriptional response in mice on high-fat diet treated with metformin was largely ablated by AMPK deficiency under the conditions examined, indicating the essential role of this kinase and its targets in metformin action in vivo.


Impact of coprophagy prevention on the growth performance, serum biochemistry, and intestinal microbiome of rabbits.
Coprophagy plays a vital role in maintaining growth and development in many small herbivores. Here, we constructed a coprophagy model by dividing rabbits into three groups, namely, control group (CON), sham-coprophagy prevention group (SCP), and coprophagy prevention group (CP), to explore the effects of coprophagy prevention on growth performance and cecal microecology in rabbits. Results showed that CP treatment decreased the feed utilization and growth performance of rabbits. Serum total cholesterol and total triglyceride in the CP group were remarkably lower than those in the other two groups. Furthermore, CP treatment destroyed cecum villi and reduced the content of short-chain fatty acids (SCFAs) in cecum contents. Gut microbiota profiling showed significant differences in the phylum and genus composition of cecal microorganisms among the three groups. At the genus level, the abundance of Oscillospira and Ruminococcus decreased significantly in the CP group. Enrichment analysis of metabolic pathways showed a significantly up-regulated differential metabolic pathway (PWY-7315, dTDP-N-acetylthomosamine biosynthesis) in the CP group compared with that in the CON group. Correlation analysis showed that the serum biochemical parameters were positively correlated with the abundance of Oscillospira, Sutterella, and Butyricimonas but negatively correlated with the abundance of Oxalobacte and Desulfovibrio. Meanwhile, the abundance of Butyricimonas and Parabacteroidesde was positively correlated with the concentration of butyric acid in the cecum. In summary, coprophagy prevention had negative effects on serum biochemistry and gut microbiota, ultimately decreasing the growth performance of rabbits. The findings provide evidence for further revealing the biological significance of coprophagy in small herbivorous mammals.


Elemental speciation studies--new directions for trace metal analysis.
Elemental speciation is reviewed as a new approach to trace metals analysis. The importance of metal speciation relative to total metal analysis is considered. Various types of chromatographic separation methods using inductively coupled plasma-mass spectrometry (ICP-MS) detection are discussed. A brief introduction to the instrumental techniques is given, as well a discussion of coupling the instrumentation. Various chromatographies are included with an emphasis on liquid chromatography (LC). Gas chromatography (GC) and capillary electrophoresis (CE) are also discussed. Application examples contrast and compare the advantages and disadvantages of ICP- MS with other detection methods. Summary tables are included for two of the LC methods.


Chemotherapy for relapsed and resistant acute nonlymphoblastic leukemia. Effect of ATA, an amsacrine-containing regime.
Twenty-nine evaluable patients with acute nonlymphoblastic leukemia (ANLL), either in relapse or resistant to initial induction therapy (ara C, daunorubicin + etoposide), received the ATA regime consisting of 100 mg/m2 per day Ara C by i.v. infusion for 4-5 days, 100 mg/m2 per day thioguanine orally for 4-5 days, and 100 mg/m2 per day amsacrine i.v. for 2-5 days. Each patient received 1-6 courses (median, 2) of the regime. There were 7 (24%) complete responders, and their duration of responses were 2, 2, 2, 5, 9+, 19, and 24+ months. The complete remission (CR) rate of patients who had a previous CR beyond 6 months (6/13, 46%) was significantly better (X2 = 4.25, p less than 0.05) than that of those who had previously relapsed within 6 months or were refractory to primary induction chemotherapy (1/16, 6%). The two groups of patients had similar patterns of treatment failure. Myelosuppression was the major toxic side effect, and nonhematological toxicities were mild and acceptable.


Abdominal wall sinus due to impacting gallstone during laparoscopic cholecystectomy: an unusual complication.
During laparoscopic cholecystectomy, perforation of the gallbladder can occurs in < or = 20% of cases, while gallstone spillage occurs in < or = 6% of cases. In most cases, there are no consequences. Gallstones can be lost in the abdominal wall as well as the abdomen during extraction of the gallbladder. The fate of such lost gallstones, which can lead to the formation of an abscess, an abdominal wall mass, or a persistent sinus, has not been studied adequately. Herein we report the case of a persistent sinus of the abdominal wall after an emergent laparoscopic cholecystectomy in an 82-year-old woman with gangrenous cholecystitis and perforation of the friable wall in association with an empyema of the gallbladder. The culture of the obtained pus was positive for Escherichia coli. After a small leak of dirty fluid from the wound of the epigastric port site of 4 months' duration, surgical exploration under local anesthesia revealed that the sinus was caused by spilled gallstones impacting into the abdominal wall between the posterior sheath and left rectus abdominalis muscle. The removal of the stones resulted in complete healing. Long-term complications after laparoscopic cholecystectomy involving the abdominal wall are rare but important possible consequences that could be avoided.


Two new polymers as candidates for rhinoplasty allografts: an experimental study in a rabbit model.
This study was performed to evaluate the biocompatibility and tensile strength of two new polymeric materials--a polyfluoro ether-modified thermoplastic polyurethane urea and a polydimethyl silicone elastomer--in an experimental rabbit model. The two polymers were implanted inside separate subperichondrial pockets created over the auricular cartilages of 12 rabbits. A control pocket received no implant. After 3 months, the animals were painlessly sacrificed, and each site was analyzed histologically for vascular congestion, acute and chronic inflammation, and fibrosis in the tissue surrounding the implant materials. There were no statistically significant differences in vascular congestion, fibrosis, or acute or chronic inflammation between the control sites and either implant site. These results suggest that the polymers are well accepted by the tissue and remain stable during the entire study period, and that they could be very suitable materials for use in nasal reconstruction.


Coronary and peripheral stenting in aorto-ostial protruding stents: The balloon assisted access to protruding stent technique.
Treatment of aorto-ostial in-stent restenosis lesions represents a challenge for interventional cardiologists. Excessive protrusion of the stent into the aorta may lead to multiple technical problems, such as difficult catheter reengagement of the vessel ostium or inability to re-wire through the stent lumen in repeat interventions. We describe a balloon assisted access to protruding stent technique in cases where conventional coaxial engagement of an aorto-ostial protruding stent with the guide catheter or passage of the guide wire through the true lumen is not feasible. This technique is applicable both in coronary and peripheral arteries.


Intensity-Based Assessment of Microbubble-Enhanced Ultrasonography: Phase-Related Diagnostic Ability for Cellular Differentiation of Hepatocellular Carcinoma.
This prospective study aimed to elucidate the effect of phase-related quantitative parameters of contrast-enhanced ultrasound (CEUS) with perflubutane microbubble agent to assess the cellular differentiation of hepatocellular carcinoma (HCC). Intensity was analyzed in 94 lesions (19.4 ± 4.9 mm, 86 patients), 47 well-differentiated HCCs (wHCCs) and 47 moderately-differentiated HCCs (mHCCs): I(e) (early phase) = I(te) (tumor) - I(le) (liver), I(p) (post-vascular phase) = I(tp) (tumor) - I(lp) (liver), I(ep) = I(e) - I(p). The area under the receiver operating characteristic curve with the best cutoff value (I(e), 13.2, I(p), -4.5, I(ep), 21.3) for discriminating between wHCC and mHCC was 0.6922 for Ie, 0.7680 for Ip and 0.7925 for Iep, which indicated a significantly greater ability to differentiate between wHCC and mHCC compared with visual/qualitative assessment (early phase, 0.6170, p = 0.04; post-vascular phase, 0.6702, p = 0.01; both phases, 0.7021, p = 0.04). In conclusion, I(ep) was found to have the highest diagnostic ability, suggesting it is a promising parameter for the cellular differentiation of HCCs with CEUS.


Mercury concentrations decline with age in the fur of females of an insectivorous terrestrial mammal (Myotis lucifugus).
Methylmercury (MeHg) is a toxic form of mercury that bioaccumulates in organisms and biomagnifies through food webs. MeHg concentrations can be high in aquatic environments, and this puts high trophic-level predators who derive energy originating from aquatic environments at risk of toxic effects. Due to the potential for bioaccumulation of MeHg over an individual's life, the risk of MeHg toxicity may increase as animals age, and this risk may be especially high in species with relatively high metabolic rates. Total mercury (THg) concentrations were measured from the fur of adult female little brown bats (Myotis lucifugus) collected between 2012 and 2017 in Salmonier Nature Park, Newfoundland and Labrador. Using linear mixed-effects models, the effects of age, year, and day of capture on THg concentrations were evaluated and interpreted with AICc and multi-model inference. We expected that THg concentrations would increase with age, and that due to annual summer moulting, individuals captured earlier in the season would have lower THg concentrations than individuals captured later in the season. Contrary to expectations, THg concentrations decreased with age and date of capture did not explain any variation in concentration. Among individuals, there was a negative relationship between the initial THg concentration of an individual and the rate of change in THg concentrations with age. Using a regression analysis, we found evidence of a population-level decline in THg concentrations in fur over the 6-year study period. Overall, the results indicate that adult female bats eliminate enough MeHg from their tissues to affect a decrease in THg concentrations in their fur over time, and that young adults are potentially at the greatest risk of experiencing toxic effects from high MeHg concentrations; this could result in reduced reproductive output, and warrants further research.


Ultrasound Assessment of the Intrauterine Device.
As the use of intrauterine devices (IUDs) has expanded in the United States, both as a highly effective form of long-acting reversible contraception (LARC) and as a tool for effective management of other gynecologic conditions, effective imaging of IUDs has become increasingly important. Ultrasound is a cost-effective, accurate, safe, and accessible imaging solution for the assessment of IUDs. This article explores the role of ultrasound in evaluation of IUDs as well as the literature regarding ultrasound guidance for device placement. It also examines the evaluation and management of IUDs with a concurrent intrauterine pregnancy.


Leaf dynamics and profitability in wild strawberries.
Leaf dynamics and carbon gain were evaluated for two species of wild strawberry, Fragaria virginiana and F. vesca. Five populations on sites representing a gradient of successional regrowth near Ithaca, N.Y., U.S.A., were studied for two or three years each. A computer-based model of plant growth and CO2 exchange combined field studies of leaf biomass dynamics with previously-determined gas exchange rates to estimate carbon balances of leaves and whole plants in different environments.Leaves were produced throughout the growing season, although there was usually a decline in rate of leaf-production in mid-summer. Leaves produced in late spring had the largest area and longest lifespan (except for overwintering leaves produced in the fall). Specific Leaf Weight (SLW) varied little with time of leaf production, but differed greatly among populations; SLW increased with amount of light received in each habitat. The population in the most open habitat had the least seasonal variation in all leaf characters. F. vesca produced lighter, longer-lived leaves than F. virginiana.Simulations showed that age had the largest effect on leaf carbon gain in high-light environments; water stress and temperature had lesser effects. Leaf carbon gain in lowlight environments was relatively unaffected by age and environmental factors other than light. Leaves in high-light environments had the greatest lifetime profit and the greatest ratio of profit to cost. Increasing lifespan by 1/3 increased profit by 80% in low-light leaves and 50% in high-light leaves. Increasing the number of days during which the leaf had the potential to exhibit high photosynthetic rate in response to high light led to little change in profit of low-light leaves while increasing profit of high-light leaves by 49%.


The radiological assessment of body fat.
A radiological method for assessment of obesity is described. A radiograph of the soft tissues of the upper arm is obtained and the subcutaneous fat layer measured. The results obtained by this method have been compared with those using a skinfold caliper and with the ponderal index, calculated from the height and weight of the subject. A highly significant correlation was found. The radiological method has several advantages: it leaves a permanent record, it is accurate in obese persons when the skinfold caliper cannot be used, and unlike the ponderal index it is not influenced by weight increases due to causes other than fat accumulation.


Pancreatic Carcinoma Causing Schnitzler's Metastasis.
Schnitzler's metastasis occurs due to the deposition of the tumor cells in the submucosa of the rectum, leading to rectal stenosis. We present a 60-year-old female who presented with abdominal pain, distension, and vomiting. Abdominal examination showed a distended abdomen and palpable bowel loops, and per rectal examination showed rectal stenosis. Imaging studies suggest rectal stenosis with carcinoma of the pancreas head. The patient was diagnosed with Schnitzler's metastasis with carcinoma of the pancreas head, which has not been reported in the literature. The patient underwent a diversion sigmoid colostomy and was planned for palliative chemotherapy after stenting the common bile duct.


Calcium-modulated membrane guanylate cyclase in synaptic transmission?
Rod outer segment guanylate cyclase 1 (ROS-GC1) is a pivotal enzyme for vertebrate phototransduction and the systematically growing evidence point to its connection with processes other than phototransduction within and outside the retina. ROS-GC1 activity is regulated by Ca2+ in two opposite modes. This regulation is indirect and occurs through Ca+-binding proteins. At nanomolar Ca2+ concentrations, ROS-GC1 is activated by GCAPs and at micromolar Ca2+-concentrations, by S100beta and neurocalcin. The former mode operates in phototransduction and the latter was proposed to play a role in synaptic activity. The last possibility was supported by findings of ROS-GC1 expression not only in various retinal layers other than photoreceptor outer segments but also outside the retina, in pineal gland and olfactory bulb. If ROS-GC1 indeed is to play a role in neurotransmission its expression must be colocalized with its Ca2+-dependent regulators and with possible targets of an increased cyclic GMP concentration, cyclic nucleotide-gated channels or cyclic GMP-dependent protein kinase, in synaptic regions. In this review these aspects of ROS-GC1 expression in retina, pineal gland and olfactory bulb are discussed.


State of the Art Review: Emerging Therapies: The Use of Insulin Sensitizers in the Treatment of Adolescents with Polycystic Ovary Syndrome (PCOS).
PCOS, a heterogeneous disorder characterized by cystic ovarian morphology, androgen excess, and/or irregular periods, emerges during or shortly after puberty. Peri- and post-pubertal obesity, insulin resistance and consequent hyperinsulinemia are highly prevalent co-morbidities of PCOS and promote an ongoing state of excess androgen. Given the relationship of insulin to androgen excess, reduction of insulin secretion and/or improvement of its action at target tissues offer the possibility of improving the physical stigmata of androgen excess by correction of the reproductive dysfunction and preventing metabolic derangements from becoming entrenched. While lifestyle changes that concentrate on behavioral, dietary and exercise regimens should be considered as first line therapy for weight reduction and normalization of insulin levels in adolescents with PCOS, several therapeutic options are available and in wide use, including oral contraceptives, metformin, thiazolidenediones and spironolactone. Overwhelmingly, the data on the safety and efficacy of these medications derive from the adult PCOS literature. Despite the paucity of randomized control trials to adequately evaluate these modalities in adolescents, their use, particularly that of metformin, has gained popularity in the pediatric endocrine community. In this article, we present an overview of the use of insulin sensitizing medications in PCOS and review both the adult and (where available) adolescent literature, focusing specifically on the use of metformin in both mono- and combination therapy.


Postoperative enteral nutrition improves survival after orthotopic small bowel transplantation in mice.
The aim of this study was to assess the effect postoperative enteral nutrition support on the survival after orthotopic small bowel transplantation in mice. A model of orthotopic small bowel transplantation used C57BL/6 mice as both donors and recipients. We demonstrated that postoperative ileus was the deciding complication after transplantation. Postoperative enteral nutritional support resolved this problem and significantly improved survival and growth rates after transplantation. These data suggest the importance of routine nutritional support after orthotopic small bowel transplantation in mice.


The influence of the supervision of psychotherapy upon being therapeutic. I: Introduction and background to the supervisory relationship.
In individual psychotherapy which derives from psychoanalysis, the most important teaching tool is individual supervision. This is in keeping with the fact that close person-toperson contact between student and teacher is the most effective mode of teaching in a university. The supervisor's first task is to transmit a body of knowledge, about psychotherapy, to the student. His second task is to give the student the benefit of his experience around clinical problems or questions. His third task is to assist the student in the development of certain attitudes which are valuable and even essential for a psychotherapist. This latter task is accomplished largely via "reality identification".


Trapping the fast-refolding state of ribonuclease A at subzero temperatures.
Unfolded ribonuclease A consists of a mixture of fast- and slow-refolding species. It is generally accepted that the slow-refolding states arise from isomerization of proline residues. We show that unfolding at subzero temperatures may be used to trap the fast-refolding species Uf, since the rate of proline isomerization slows down at a much faster rate than protein unfolding. The unfolding was carried out in 5 M guanidine hydrochloride; at -15 degrees C the protein unfolding process is complete within 30 s and under these conditions there is less than 1.5% proline isomerization. By using ribonuclease in which Tyr-115 was nitrated it was possible to rule out significant isomerization of Pro-114 in the observed slow-unfolding step.


Professional involvement in self-help groups for parents of high-risk newborns.
Patterns of professional involvement and control in self-help groups were studied through a questionnaire completed by representatives of 63 self-help groups for parents of high-risk newborns located throughout the United States. Results indicated that when professionals began a group, they were likely to continue to maintain control over it. Groups controlled by professionals were more likely to utilize professional counseling as a form of support and less likely to provide outreach services to parents following hospital discharge of their infants. Group stability, growth, and innovation were associated with more consultation between parents and professionals in the planning process.


Recombinant virus-like particle presenting a newly identified coxsackievirus A10 neutralization epitope induces protective immunity in mice.
Coxsackievirus A10 (CVA10) has emerged as one of the major pathogens of hand, foot, and mouth disease in recent years. However, there are no approved vaccines or effective drugs against CVA10. Several experimental CVA10 vaccines have been shown to elicit neutralizing antibodies that could confer protection against viral infection. However, neutralizing antigenic sites on CVA10 capsid have not been well characterized. Here, we report the characterization of linear neutralization epitopes of CVA10 and the development of a CVA10 vaccine based on the identified epitopes. We showed that peptide VP2-P28, corresponding to residues 136 to 150 of VP2, were recognized by anti-inactivated CVA10 sera and effectively inhibited anti-CVA10 sera-mediated neutralization, suggesting that this peptide contains neutralizing epitopes. Insertion of VP2-P28 into hepatitis B core antigen (HBc) resulted in a chimeric virus-like particle (VLP; designated HBc-P28) with the CVA10 epitope exposed on the particle surface. HBc-P28 VLP elicited strong antibody responses against VP2-P28 in mice. Anti-HBc-P28 sera could neutralize both CVA10 clinical isolates and prototype strain, consistent with the fact that the VP2-P28 sequence is highly conserved among CVA10 strains. In addition, anti-HBc-P28 sera failed to cross-neutralize other HFMD-causing enteroviruses, indicating that neutralizing antibodies elicited by HBc-P28 VLP were CVA10-specific. Importantly, anti-HBc-P28 sera were able to provide efficient protection against lethal CVA10 infection in recipient mice. Collectively, these data show that peptide VP2-P28 represents a CVA10-specific linear neutralizing antigenic site and chimeric VLP displaying this peptide is a promising epitope-based CVA10 vaccine candidate.


Granger causality--statistical analysis under a configural perspective.
The concept of Granger causality can be used to examine putative causal relations between two series of scores. Based on regression models, it is asked whether one series can be considered the cause for the second series. In this article, we propose extending the pool of methods available for testing hypotheses that are compatible with Granger causation by adopting a configural perspective. This perspective allows researchers to assume that effects exist for specific categories only or for specific sectors of the data space, but not for other categories or sectors. Configural Frequency Analysis (CFA) is proposed as the method of analysis from a configural perspective. CFA base models are derived for the exploratory analysis of Granger causation. These models are specified so that they parallel the regression models used for variable-oriented analysis of hypotheses of Granger causation. An example from the development of aggression in adolescence is used. The example shows that only one pattern of change in aggressive impulses over time Granger-causes change in physical aggression against peers.


Predictors of weight loss success. Exercise vs. dietary self-efficacy and treatment attendance.
Pre-treatment diet and exercise self-efficacies can predict weight loss success. Changes in diet self-efficacy across treatment appear to be even stronger predictors than baseline levels, but research on changes in exercise self-efficacy is lacking. Using data from a pilot study evaluating tangible reinforcement for weight loss (N=30), we examined the impact of changes in diet and exercise self-efficacy on outcomes. Multiple regression analyses indicated that treatment attendance and changes in exercise self-efficacy during treatment were the strongest predictors of weight loss. Developing weight loss programs that foster the development of exercise self-efficacy may enhance participants' success.


Low dose- high frequency, case based psychomotor CPR training improves compression fraction for patients with in-hospital cardiac arrest.
High quality cardiopulmonary resuscitation (CPR) is critical to improve survival from cardiac arrest. While low dose- high frequency case-based training enhances CPR skill retention, it is unclear if this training method is feasible in a clinical environment and if it yields improved clinical CPR quality during in-hospital cardiac arrest. We evaluated the implementation of a novel platform providing low dose- high frequency psychomotor CPR training and its impact upon CPR quality. The described training platform was launched on two nursing units (60 beds) in a university teaching hospital. Quarterly utilization of the platform was integrated into normal clinical duties of hospital staff. Simulated CPR performance and staff compliance were evaluated pre- and post-intervention. In addition, clinical CPR performance was evaluated for periods of six months before and after four quarters of implementation (median, IQR). The low dose, high frequency CPR training led to retention of simulated CPR skills (compression rate, depth and fraction) during each quarter exceeding high-quality guideline thresholds. Clinical CPR quality, measured by compression fraction (Pre: 83% (73, 95) and Post: 93% (88, 98), p < 0.001) and rate (Pre: 109 (96, 126) and Post: 120 (108, 130), p = 0.008) increased significantly following platform implementation. Over the intervention period, program compliance was greater than 97%. Low dose-high frequency case based psychomotor CPR training is feasible in a clinical setting with high compliance. In two nursing units, this method of training resulted in enhanced CPR skill retention and improved in-hospital clinical CPR quality.


A rare case of ventricular septal defect associated with patent ductus arteriosus and coarctation of aorta: a multimodality approach.
This case report describes a unique form of a complex congenital anomaly; ventricular septal defect associated with patent ductus arterious and coarctation of aorta in a 21-year-old patient with a multimodality approach.


Diffusion-weighted imaging in kainic acid-induced complex partial status epilepticus in dogs.
To investigate diffusion-weighted imaging (DWI) in status epilepticus, a canine model of kainic acid (KA)-induced complex partial status epilepticus (CPSE) was produced. In order to validate its usefulness, MR imaging was carried out at various times following onset of CPSE followed by histopathology. Six normal dogs were used in this study. In each dog, a cannula was stereotactically inserted into the left amygdala. One week after surgery, all dogs were imaged at MRI. Pre-injection imaging consisted of T2 weighted (T2W) imaging, fluid attenuated inversion recovery (FLAIR), and DWI. Two weeks after surgery, five dogs received intraamygdaloid KA microinjections. One dog was used as a control. MRI was carried out at 3, 6, 12, 24 and 48 h after onset of CPSE. Animals were euthanized immediately after MRI for histopathological evaluation. The average of each apparent diffusion coefficient (ADC) in the regions of interest was calculated from each DWI. At 3 and 6 h, DWI hyperintensity and low ADC were found in the injected amygdala, without any T2W and FLAIR imaging changes. At 12 and 24 h, all imaging showed hyperintensity with higher ADC in the amygdala and the hippocampus. At 48 h, all imaging techniques showed continued hyperintensity, but ADC showed a trend towards normalization. This increasing hyperintensity in DWIs were in agreement with the degree of histopathology during CPSE. This study suggests that DWI is a useful imaging method for finding the epileptic focus or for examining potential epileptic brain damage in status epilepticus.


Chronic atrophic gastritis and Helicobacter pylori infection status in liver transplant recipients.
To investigate the abnormalities of the upper gastrointestinal tract in liver transplant (LT) recipients, especially the prevalence of Helicobacter pylori infection and the incidence of chronic atrophic gastritis (CAG), and to explore the efficacy and safety of H pylori eradication treatment. Endoscopic screening was performed prospectively on LT recipients who received regular follow-up in our center. A group of healthy subjects with same age and sex was selected as a control group at a ratio of 1:3 with propensity score matching. All H pylori-positive recipients received Bismuth-containing quadruple therapy (esomeprazole 20 mg + clarithromycin 500 mg + amoxicillin 1 g + bismuth 220 mg, all of the medicines were applied twice daily, for 14 days). The prevalence of H pylori infection was significantly lower in LT group than control group [12/102 (11.8%) vs 98/306 (32.0%), P < .001], whereas the prevalence of CAG was similar between the two groups [48/102 (47.1%) vs 138/306 (45.1%), P = .731]. Meanwhile, the incidence of reflux esophagitis [18/102 (17.6%) vs 31/306 (10.1%), P = .043] and bile regurgitation [19/102 (18.6%) vs 30/306 (9.8%), P = .018] were higher in LT group. No correlation between the incidence of upper gastroduodenal abnormalities and postoperative time after liver transplantation was found. The success rate of H pylori eradication therapy was 100% (10/10). The blood concentration of immunosuppressants was 1.7-3.6 times above baseline values during H pylori eradication therapy; however, no severe adverse effects were observed during the proceed with dose adjustments of the immunosuppressants. Although the prevalence of H pylori infection was lower in LT recipients than in control subjects, the prevalence of CAG was like that of the general population. H pylori eradication therapy was safe and effective after liver transplantation in our preliminary study.


Antiproliferative efficacy of the third-generation bisphosphonate, zoledronic acid, combined with other anticancer drugs in leukemic cell lines.
Bisphosphonates are widely used to treat bone diseases and appear to possess antitumor activity. Moreover, we recently found that a third-generation bisphosphonate, zoledronic acid (ZOL), synergistically interacts with imatinib in vitro and in vivo to induce antileukemic activity, and others have reported that ZOL interacts synergistically with paclitaxel. Thus, the efficacy of other antileukemic agents combined with ZOL should be evaluated experimentally. In this study, we investigated the effects of concurrent and sequential combinations of ZOL with several commonly used antileukemic agents, including imatinib, on the in vitro growth of leukemia cell lines. As a complement to our previous finding that ZOL synergistically augments the effects of imatinib, we report here that ZOL acts additively when administered concurrently with hydroxyurea (HU), cytarabine (Ara-C), or daunorubicin (DNR) in some leukemic cell lines. Furthermore, one day of ZOL pretreatment augmented the sensitivity of imatinib and Ara-C. Therefore, concurrent or sequential administration of ZOL with imatinib, HU, Ara-C, or DNR may increase the efficacy of leukemia treatment.


Somatosensory activity relevant for motor output. A summary of presentations and discussion.
Several aspects of the function of receptors which contribute to somatic sensations are reviewed. First, there is evidence for a role of large-diameter cutaneous afferents in the reflex regulation of precision movements by the hand. Second, large-diameter muscle afferents from the intrinsic muscles of the hand, probably from primary muscle spindle afferents, can evoke specific sensations of finger movement. Third, the variable relationship between discharges in human C fibers from the hand and the specific sensation of pain is investigated. Activity in large-diameter cutaneous afferents can probably modify this sensation. Finally, the properties of small-diameter afferent fibers innervating joints are shown to be consistent with a role in the reflex regulation of joint integrity.


Non-clinical vaccine safety assessment.
As vaccines are undoubtedly classified as pharmaceuticals, they have to be submitted to strict non-clinical safety evaluation. The context of their prophylactic use requires that every effort is made to ensure their safe use. Their safety evaluation is complex as they act through a multistage mechanism in which the vaccine by itself acts as a pro-drug, antibodies and activated lymphocytes being the actual effectors. Therefore, several potential toxicities must be considered: direct toxicity of the test article, toxicity linked to the pharmacodynamic activity of the vaccine, activation of pre-existing disorders, toxicity of contaminants and impurities and other adverse reactions due to interaction between the various components. Guidelines dealing with vaccines include general guidelines applicable to all pharmaceuticals, such as ICH S6, and also more specific documents which allow some flexibility in study design. Among the various studies, if single-dose studies are generally part of the quality control test battery, repeated dose studies are pivotal. The animal model and treatment schedule selection and the parameters investigated are critical for the relevance of this safety assessment. Immunological and safety pharmacology parameters should be adapted to the specific properties of vaccines and added to this type of study. Vaccines intended for pregnant women or women of child-bearing age require embryo-fetal and post-natal studies with an adapted design to obtain appropriate fetal and maternal exposure during gestation with continuation into the post-natal period. Tests exist to detect hypersensitivity or autoimmune reactions, but require further validation. In addition to this tailor-made approach, any adjuvant or active component added to the vaccine formulation necessitate their own assessment using studies routinely performed for new drugs. From this review, vaccine toxicology would appear to be a separate discipline on its own whose predictivity will be increased by new method development.


Antibodies against type II phospholipase A2 prevent renal injury due to ischemia and reperfusion in rats.
This study was performed to determine the involvement of type II phospholipase A2 (PLA2-II) in renal injury caused by ischemia and reperfusion. Ischemia and reperfusion significantly elevated levels of blood urea nitrogen and serum creatinine in rats. These increases were significantly reduced by i.v. administration of rabbit IgG F(ab')2 fragments against rat PLA2-II. Increased levels of acid-stable PLA2 activity in the kidney were caused by ischemia and reperfusion, and were suppressed by administration of anti-PLA2-II F(ab')2. Increased levels of myeloperoxidase activity, a marker of neutrophil infiltration, in the kidney were also reduced after anti-PLA2-II F(ab')2 treatment. These results suggest that PLA2-II plays a pivotal role in pathogenesis of ischemia and reperfusion injury through induction of neutrophil infiltration.


Hemoperfusion with HA380 in acute type A aortic dissection patients undergoing aortic arch operation (HPAO): a randomized, controlled, double-blind clinical trial.
Cardiopulmonary bypass (CPB) is an important cause of significant systemic inflammatory response syndrome (SIRS) in the surgical treatment of acute type A aortic dissection (ATAAD). In patients with arch vessel involvement, extensive surgical repairs often necessitate prolonged use of CPB and results in extensive inflammatory responses. Cytokines and chemokines released during CPB contribute to the progression of SIRS, increase perioperative complications, and negatively impact surgical outcomes. A cytokine adsorber (HA380) is expected to reduce the level of cytokines during CPB, which may decrease both intraoperative and postoperative inflammation. The purpose of this study is to investigate if HA380 is able to reduce the levels of inflammatory cytokines and decrease perioperative complications in ATAAD patients undergoing CPB and deep hypothermic circulatory arrest (DHCA). This study is a single-center, randomized, controlled, double-blind clinical trial. The study aims to recruit 88 patients with ATAAD and aortic arch involvement who will undergo CPB and DHCA to repair the dissected aorta. Patients will be randomized equally into the CPB/DHCA only group (control group) and the CPB/DHCA + HA380 hemoperfusion group (intervention group), with 44 patients each. Patients in the control group will undergo CPB and DHCA only, while patients in the intervention group will undergo continuous hemoperfusion with HA380, in addition to CPB and DHCA. The primary outcome is a composite of major perioperative complications. The secondary outcomes include related inflammatory markers, coagulation parameters, and minor perioperative complications. To comprehensively evaluate the effect of hemoperfusion on the perioperative outcomes, we will also determine if there are differences in perioperative all-cause mortality, length of ICU stay, and total hospitalization costs. In the current trial, hemoperfusion will be applied in patients undergoing CPB and DHCA for repair of the aorta involving the aortic arch. This trial aims to test the safety and efficacy of our hemoperfusion device (HA380) in such settings. Upon completion of the trial, we will determine if HA380 is effective in reducing perioperative proinflammatory cytokine levels. Further, we will also verify if reduction in the proinflammatory cytokine levels, if present, translates to improvement in patient outcomes. ClinicalTrials.gov NCT04007484 . Registered on 1 July 2019 (retrospectively registered).


Differentiation of Salmonella vaccinated and infected animals by serological detection of antibody to T3SS effector SsaK in an indirect ELISA.
The differentiation of animals that are vaccinated and those that are naturally infected with Salmonella is difficult by conventional serological tests. We have shown here an indirect Enzyme-linked immunosorbent assay for detection of Salmonella infection based on the presence of a Type III secretory effector SsaK in the sera.


Assessing Quality of Life for Pets with Dermatologic Disease and Their Owners.
Quality of life (QoL) is defined as "the degree to which an individual enjoys his or her life." In canine dermatology, 2 research groups have developed and validated questionnaires for QoL assessment in dogs with dermatologic conditions, the first being limited to canine atopic dermatitis, and the second one applicable to all skin conditions. A third group is working on the validation of a different QoL questionnaire coupled with a treatment success assessment tool. In general, there is an inverse correlation between QoL and pruritus.


Treatment of a case of thromboembolism resulting from thoracic outlet syndrome with intra-arterial urokinase infusion.
A 36-year-old man with thoracic outlet syndrome, admitted to the hospital with digital ischemia from subclavian artery thrombosis and distal embolization, was given intra-arterial urokinase. Thrombus in the subclavian artery was lysed successfully and peripheral emboli were partially cleared, resulting in relief of digital symptoms. Although surgical decompression and vascular reconstruction at the thoracic outlet may be necessary, this technique provides a means of recanalizing small distal vessels.


What influences the outcome of valve replacement in critical aortic stenosis?
Even patients with severely reduced left ventricular function and critical aortic stenosis can improve notably following valve replacement.


Clinical approach to the diagnostic evaluation of progressive neuromuscular diseases.
This article has reviewed the clinical approach to the diagnostic evaluation of progressive neuromuscular diseases with an emphasis on relevant neuromuscular history, family history, clinical examination findings, laboratory studies, and a brief discussion of the role of muscle biopsy. Molecular genetic and immunocytochemistry studies of muscle have been major advances in the diagnostic evaluation of the neuromuscular disease patient; however, all diagnostic information must be interpreted within the context of relevant clinical information. In some instances, a precise diagnosis is not medically possible; however, the accurate characterization of an individual patient within the most appropriate NMD clinical syndrome often allows the clinician to provide the patient and family with accurate prognostic information and anticipatory guidance for the future. After synthesizing all available clinical and diagnostic information, the physiatrist or neurologist may at times determine that an NMD patient has an inappropriate diagnosis warranting further diagnostic evaluation. This issue focuses on the rehabilitation of progressive neuromuscular diseases with an emphasis on optimization of health, prevention or minimization of complications, and enhancement of quality of life. Appropriate rehabilitation approaches require an accurate diagnosis. In addition, patient quality of life in NMD depends on access to current and accurate information. The first step in providing accurate information and appropriate treatment is constantly ensuring that NMD patients have appropriate diagnoses based on a through evaluation of clinical information and appropriate application of current medical science and available diagnostic technology.


Influence of some inflammatory or antiinflammatory substances on skin biopsies investigated with microcalorimetry and respiratory techniques.
Thermal effects were achieved from skin biopsies (epidermis, corium, subcutis) measured by batch-microcalorimetry, after exposure to potent inflammatory or antiinflammatory substances (compound 48/80, histamine, serotonine and hydrocortisone phosphate). Exothermic effects were achieved from 48-80, histamine and hydrocortisone phosphate, when exposed to the skin and lasting for varying time. From serotonine initial endothermic signals were achieved but after about 50 min there was stabilizing of the thermal reaction at the baseline. To analyze the nature of these non-specific thermal effects, determinations of O2 consumption and CO2 production were performed, using Warburg technique on exactly matched tissue pieces exposed to the same substances and run parallely with the microcalorimetrical determinations. An increased respiration, probably as an indicator of increased metabolism was registrated for all test substances acting on the skin. It felt, that the thermal effects reflect an increased metabolism of the tissue exposed to the test substances.


Evaluation of the solvent black 3 fingermark enhancement reagent: part 1 - investigation of fundamental interactions and comparisons with other lipid-specific reagents.
The fundamental interactions between sebaceous constituents of fingermarks and three lipid specific fingermark enhancement reagents (solvent black 3, basic violet 3 and basic violet 2) are reported. The staining of fingermarks is investigated using optical microscopy, and the interaction of the reagents with individual constituents is explored using spot tests. It is demonstrated that solvent black 3, basic violet 3 and basic violet 2 all interact with different constituents of sebaceous sweat, and this may offer potential for using the reagents in sequence for fingermark enhancement. Further tests to explore the effect of dye concentration on reagent effectiveness indicate that dye concentration can be reduced by up to 25% without significant detriment to effectiveness. It is shown that there is little practical difference between solvent black 3 formulations with the solvents (ethanol and 1-methoxy-2-propanol) used in this study. The study also indicates that basic violet 2 may have some operational advantages over basic violet 3 and may be worthy of further investigation.


Cisplatin induced cell killing and chromosomal aberrations in CHO cells: treated during G1 or S phase.
Variation in sensitivity to cisplatin during the cell cycle was studied in synchronous CHO cells treated during G1 or late S phase. The cells were assayed for cell killing, cell-cycle delay, and chromosomal aberrations after they were treated with cisplatin (1-12 micrograms/ml) for 1 h at 37 degrees C. They were either plated for colony survival, or colcemid was added from 12 to 40 h after plating followed by fixation 4 h later for analysis of chromosomal aberrations after the cells completed 1 or 2 cycles (i.e. first or second mitosis). Cells treated with 6 micrograms/ml exhibited about a 10-h delay during the first cycle after treatment during G1 compared with about 3 h during the first cycle and 6 h during the second cycle after treatment during late S. In both cases, cells entering metaphase exhibited predominantly chromatid-type breaks and exchanges. For both cell killing and chromosomal aberrations, the cells in G1 were 1.5-1.6 times more sensitive than those treated in late S, with 1 aberration per cell corresponding to about 37% survival. However, the exchanges and breaks were observed primarily in the first mitosis when cells were treated in G1 compared with the second mitosis when cells were treated in late S. These results suggest that DNA replication opposite cisplatin cross-links in the DNA results in lethal chromosomal aberrations.


Hemophagocytic Lymphohistiocytosis: Early Treatment Leading to an Excellent Outcome.
HLH is a rare and deadly disease increasingly more present in adults, but following treatment protocol may yield favorable results.


Advances in understanding bone cancer pain.
Experimental animal models of bone cancer pain have emerged and findings have provided a unique glimpse into unraveling the mechanism that drives this debilitating condition. Key contributors to the generation and maintenance of bone cancer pain are tumor-induced osteolysis, tumor itself, and production of nociceptive mediators in the bone-tumor microenvironment.


Computer-generated model surgery. An exacting approach to complex craniomaxillofacial disharmonies.
Description and applications are presented for a technique of craniomaxillofacial-model surgery. The technique utilizes models created by and based on computer technology. This paper is based on the experience derived from 22 cases in which computer-generated models were used as the means of evaluation, presurgical planning and practice.


Shoulder dystocia: a fetal-physician risk.
Trauma that occurs as a result of shoulder dystocia is an important cause of neonatal morbidity. If the occurrence of severe shoulder dystocia, resulting in fetal asphyxia and trauma, could be accurately predicted from maternal risk factors, then a cesarean section would be indicated to prevent the poor outcome. The information available in the obstetric literature, however, is contradictory regarding whether shoulder dystocia can be predicted. In the present study, the patients at greatest risk of shoulder dystocia (all 394 mothers delivering neonates with birth weights greater than or equal to 4000 gm over a 2-year period) were examined. A three-way discriminant analysis was used to determine if a model could be developed that could effectively predict those patients who would be included in each of the groups of no shoulder dystocia, shoulder dystocia without trauma (29 patients), and shoulder dystocia with trauma (20 patients). Three factors, including birth weight, prolonged deceleration phase, and length of second stage labor, were found individually to contribute significantly to the classification. However, when examined in detail, it was noted that while 94% of cases with no shoulder dystocia would be detected, only 16% of the cases of shoulder dystocia with trauma would be predicted by this model. We conclude that in the group of pregnancies delivering neonates greater than or equal to 4000 gm, the occurrence of shoulder dystocia cannot be predicted from clinical characteristics or labor abnormalities, and that the occurrence of shoulder dystocia is not evidence of medical malpractice.


Significant reduction in post-lumbar puncture headache using an atraumatic needle. A double-blind, controlled clinical trial.
A characteristic headache occurs in about 36-55% of patients after lumbar puncture, and many of these patients need bed rest for one or more days to get relief. In a double-blind randomized trial we compared a new 22-gauge atraumatic puncture needle with the most widely used 20-gauge "conventional" needle. In 49 patients randomized to the atraumatic needle, post-lumbar puncture headache occurred in 6%, whereas in the 50 patients randomized to the conventional needle this occurred in 32% (p = 0.001). On the basis of these results we recommend use of the atraumatic needle in order to diminish the frequency of post-lumbar puncture headache.


Removing race and body surface area indexation for estimated kidney function based drug dosing: Aminoglycosides as justification of these principles.
The use of race in medicine can contribute to health inequity. Updated equations for estimated glomerular filtration rate (eGFR) without race have been published. Likewise, de-indexation of eGFR to body surface area (BSA) has been recommended by regulatory guidance for drug dosing in renal impairment. Clinical data justifying these recommendations for drug dosing are sparse. We examined the gain or loss of precision in drug dosing with estimated creatinine clearance (eCLcr) and eGFR using serum creatinine (eGFRcr) with and without race and BSA indexation by evaluating the population pharmacokinetics of the aminoglycosides as a classic drug class to probe kidney function. Medical records from adult patients treated with gentamicin or tobramycin over a 13-year period were queried. Population pharmacokinetic analyses were performed using a 1-compartment base structural model. Models compared body size descriptors as covariates of the volume of distribution (V). Estimated creatinine clearance and eGFRcr using multiple contemporary equations with and without BSA indexation were tested as covariates of clearance (CL). The final data set included 2968 patients treated with either gentamicin (20.2%) or tobramycin (79.8%). Patients self-identified as Caucasian (82%), African-American (10%), or other. The median [5th, 95th percentile] weight and BSA were 80.5 [49.4, 136] kg and 1.94 [1.48, 2.56] m2 , respectively. Models of eCLcr and eGFRcr without indexation to BSA had a better model fit than eGFRcr indexed to BSA for aminoglycoside CL. The 2021 Chronic Kidney Disease Epidemiology collaboration (CKD-EPI) eGFRcr equation (no race, no BSA indexation) provided a comparable model fit to the 2009 CKD-EPI eGFRcr equation (with race, no BSA indexation) for aminoglycoside CL. Race is not a relevant covariate of aminoglycoside CL. The 2021 CKD-EPI eGFR equation without race and BSA indexation is a better method for gentamicin and tobramycin CL estimation. Confirmation of these results for other drugs can support the harmonization of dosing by kidney function.


Red blood cell distribution width in glioblastoma.
Glioblastoma (GBM) is the most common and deadly adult brain tumor. Red blood cell distribution width (RDW) has been found in non-central nervous system neoplasms to be associated with survival. This study aims to assess the prognostic value of pre-operative RDW and trends in RDW over time during the disease course. This single-institution retrospective cohort study identified patients ≥ 18 years old with pathology-proved glioblastoma treated between April 2003-May 2017 from an institutional database. A Cox proportional hazards model was developed using known prognostic clinical variables to predict overall survival time; a second model incorporating continuously valued RDW was then created. The additional prognostic value of RDW was assessed with a joint model F-test. The variation of RDW-CV over time was evaluated with linear mixed model of RDW. A post-hoc exploratory analysis was performed to assess the trend in RDW lab value leading up to time of death. 346 adult GBM patients were identified; complete survival data was available for all patients. The addition of RDW to the multivariable Cox proportional hazards model did not increase prognostic value. There was an upward trend in RDW throughout the post-operative disease course. In a post-hoc analysis, there was an upward trend in RDW leading up to the time of death. Although RDW has been prognostic of survival for many inflammatory, prothrombotic, and neoplastic diseases, pre-operative RDW was not associated with overall survival in GBM patients. RDW trended upwards throughout the disease course, suggesting possible systemic inflammatory effects of either glioblastoma or treatment.


Inhibitory kinetics of chlorocinnamic acids on mushroom tyrosinase.
Tyrosinase (EC 220.127.116.11) is the key enzyme of most food enzymatic oxidation. Tyrosinase inhibitors are important in food industry. In the present paper, 2-chlorcinnamic acid and 2,4-dichlorocinnamic acid were synthesized and the inhibitory kinetics on mushroom tyrosinase were investigated. The results showed that both compounds synthesized could inhibit tyrosinase activity. For monophenolase activity, both chlorocinnamic acids could extended the lag time and decrease the steady-state activities, 2-chlorcinnamic acid extended the lag time just by 5%, and 2,4-dichlorcinnamic acid extended the lag time more than by 30.4%. For diphenolase activity, the IC50 values of 2-chlorcinnamic acid and 2,4-dichlorocinnamic acid were determined to be 0.765 mM and 0.295 mM, respectively. The inhibition kinetics showed that 2-chlorcinnamic acid and 2,4-dichlorocinnamic acid displayed a reversible and uncompetitive mechanism. The inhibition constants were determined to be 0.348 mM and 0.159 mM, respectively. The research may supply the basis for designing new tyrosinase inhibitors.


The Smooth Away From Expected (SAFE) non-inferiority frontier: theory and implementation with an application to the D3 trial.
In a non-inferiority trial, the choice of margin depends on the expected control event risk. If the true risk differs from expected, power and interpretability of results can be affected. A non-inferiority frontier pre-specifies an appropriate non-inferiority margin for each value of control event risk. D3 is a non-inferiority trial comparing two treatment regimens in children living with HIV, designed assuming a control event risk of 12%, a non-inferiority margin of 10%, 80% power and a significance level (α) of 0.025. We consider approaches to choosing and implementing a frontier for this already funded trial, where changing the sample size substantially would be difficult. In D3, we fix the non-inferiority margin at 10%, 8% and 5% for control event risks of ≥9%, 5% and 1%, respectively. We propose four frontiers which fit these fixed points, including a Smooth Away From Expected (SAFE) frontier. Analysis approaches considered are as follows: using the pre-specified significance level (α=0.025); always using a reduced significance level (to achieve α≤0.025 across control event risks); reducing significance levels only when the control event risk differs significantly from expected (control event risk <9%); and using a likelihood ratio test. We compare power and type 1 error for SAFE with other frontiers. Changing the significance level only when the control event risk is <9% achieves approximately nominal (<3%) type I error rate and maintains reasonable power for control event risks between 1 and 15%. The likelihood ratio test method performs similarly, but the results are more complex to present. Other analysis methods lead to either inflated type 1 error or badly reduced power. The SAFE frontier gives more interpretable results with low control event risks than other frontiers (i.e. it uses more reasonable non-inferiority margins). Other frontiers do not achieve power close (i.e. within 1%) to SAFE across the range of likely control event risks while controlling type I error. The SAFE non-inferiority frontier will be used in D3, and the non-inferiority margin and significance level will be modified if the control event risk is lower than expected. This ensures results will remain interpretable if design assumptions are incorrect, while achieving similar power. A similar approach could be considered for other non-inferiority trials where the control event risk is uncertain.


Preparation and characterisation of attenuated cold-adapted influenza A reassortants derived from the A/Leningrad/134/17/57 donor strain.
The development of a rapid cell culture method for the preparation of cold-adapted (ca) influenza A reassortant viruses is described and compared with a currently used egg method. Mixtures of the ca donor A/Leningrad/134/17/57-ca (A/Len/17) and A/Beijing/32/92 (A/Beij/32), a recent H3N2 epidemic strain, were used to co-infect chicken embryo kidney (CEK) cell cultures; reassortant progeny were selected using an infectious centre assay. The assay was capable of detecting interference where the infectivity ratio for A/Len/17 and A/Beij/32 was 1:7. Progeny viruses were characterised genetically by amplification of defined regions within each of the six internal genes by PCR and identification of the products by restriction enzyme analysis. Reassortants were also tested for ca and temperature-sensitive (ts) phenotype and the identity of the surface antigens. The infectious centre assay was shown to be an effective method for isolating reassortant progeny that possessed the haemagglutinin and neuraminidase surface antigens of A/Beij/32. Reassortants with the six internal genes derived from the donor strain and possessing the ca and ts phenotype were readily obtained when (a) an infectivity ratio of 1:49 was used and (b) two plaque-to-plaque isolations of progeny virus were made after growth at 25 degrees C and one at 34 degrees C, both in the presence of antiserum to the donor strain.


Ankyrin G and voltage gated sodium channels colocalize in human neuroma--key proteins of membrane remodeling after axonal injury.
We tested if ankyrin G could be detected in human neuroma, if it colocalized with site-specific peripheral nerve sodium channels that accumulate at axon tips of injured nerve, and if there are differences in the distribution of these proteins in non-painful neuroma and painful neuroma tissue vs. normal nerve. Frozen sections from one painful, six non-painful, and three normal nerves were immunocytochemically examined. A double labeling technique with highly specific antibodies against peripheral nerve type 1 (Na(v)1.7), and peripheral nerve type 3 (Na(v)1.8) sodium channels and anti-ankyrin G antibodies detected sodium channels and ankyrin G on the same section, using confocal laser scanning microscopy. Ankyrin G colocalized with both types of sodium channels. Neuroma specimens exhibited considerably larger immunofluorescence for both sodium channels and ankyrin G compared with normal nerve. The painful neuroma presented an even more pronounced immunolabeling in clusters. Findings support results from animal models that link ankyrin G with clustering of sodium channels at axon tips of unmyelinated, sprouting fibers. A common (repair-) mechanism that exists throughout the human nervous system for clustering sodium channels at a high density is assumed. A dysregulation in this membrane remodeling mechanism might be an initial step in a cascade that leads to a painful rather than a non-painful neuroma.


What influences graduate medical students' beliefs of lower back pain? A mixed methods cross sectional study.
Low back pain (LBP) is a common condition with substantial associated disability and costs, best understood using a biopsychosocial approach. Research demonstrates that beliefs about LBP are important, with biomedical beliefs influencing practitioner's management and patient recovery. Beliefs about LBP can be inconsistent amongst healthcare and medical students. The aim of this study was to investigate graduate medical student's beliefs of LBP and what influences them. A cross sectional mixed methods study of Phase 1 (first year) and Phase 3 (third and fourth year) current graduate medical students at the University of Warwick (MBChB) was conducted. Participants were recruited via voluntary response sampling. A survey investigated LBP beliefs, utilising the Back Beliefs Questionnaire (BBQ) and Health Care Providers' Pain and Impairment Relationship Scale (HC-PAIRS). Qualitative data was collected on what influences beliefs about the causes and management of LBP, which was analysed descriptively using thematic analysis. Fifty-seven students completed the questionnaire (61% female), with a mean age of 27.2 years. Eighty two percent of participants reported a history of LBP. Median BBQ scores were 31.5 for phase 1 and 31 for phase 3, with median HC-PAIRS scores of 57 and 60 for phase 1 and phase 3 students respectively. Three main themes emerged from the qualitative data: Sources of influence, influence of personal experience and influence of medical education. Participants discussed single or multiple sources influencing their beliefs about the causes and management of LBP. Another main theme was the influence of experiencing LBP personally or through discussions with family, friends and patients. The final main theme described the influence of medical education, including lectures, seminars and clinical placements. The HC-PAIRS and BBQ scores suggest graduate medical students in this sample tended to have positive beliefs about the outcome of LBP and functional expectations of chronic LBP patients, consistent with other healthcare students. The findings from qualitative data suggest how medical students form beliefs about the causes and management of LBP is complex.


Pain may be inevitable; inadequate management is not.
Assessing and managing pain in the critically ill patient is challenging. Reproducible and clinically applicable pain measurement scales have yet to be validated and ubiquitously applied in the intensive care unit setting. Critical care clinicians, both physicians and nurses, should thoughtfully monitor their patient's pain level, periodically reassess their practice and critically evaluate the efficacy of pharmacological and nonpharmacological analgesic interventions.


Immunochemical study of host proteins in human supragingival compared with denture plaque.
The contribution of proteins from saliva and gingival crevicular fluid (GCF) to plaque was determined by comparing extracts of supragingival plaque of unknown age formed on normal teeth with plaque formed on artificial teeth in complete or partial dentures where crevicular fluid is absent. There was a total absence of albumin and a virtual absence of IgG from denture plaque samples, confirming their crevicular origin. The concentration of lactoferrin was much higher than that of lysozyme in all supragingival but not in the denture plaque samples, suggesting that GCF provided more lactoferrin than lysozyme to plaque. Amylase was a component in both denture and supragingival plaque, present in similar amounts in both deposits. Cysteine-containing phosphoproteins from saliva were in low concentration but present in all plaque samples; proline-rich proteins were virtually absent, reflecting the high vulnerability to proteolysis of these proteins. Salivary proteins in plaque extracts do not correspond with their relative concentrations in saliva.


A case series report on successful management of patients with COVID-19-associated lymphopenia and potential application of PG2.
Lymphopenia and the resultant high neutrophil-to-lymphocyte ratio (NLR) are hallmark signs of severe COVID-19, and effective treatment remains unavailable. We retrospectively reviewed the outcomes of COVID-19 in a cohort of 26 patients admitted to Chung Shan Medical University Hospital (Taichung City, Taiwan). Twenty-five of the 26 patients recovered, including 9 patients with mild/moderate illness and 16 patients with severe/critical illness recovered. One patient died after refusing treatment. We report the cases of four patients with high NLRs and marked lymphopenia, despite receiving standard care. A novel injectable botanical drug, PG2, containing Astragalus polysaccharides, was administered to them as an immune modulator. The decrease in the NLR in these four patients was faster than that of other patients in the cohort (0.80 vs. 0.34 per day). All patients recovered from severe COVID-19 showed decreased NLR and normalized lymphocyte counts before discharge. Administration of PG2 may be of benefit to patients with moderate to severe COVID-19 and lymphopenia.


Repair of Acute Grade 3 Combined Posterolateral Corner Avulsion Injuries Using an Enhanced Fixation Technique.
Previous studies have suggested that surgical repair of the posterolateral corner (PLC) may be inferior to reconstruction. We hypothesized that acute repair (<3 weeks) of avulsion-type PLC multiligament knee injuries with no midsubstance injury would lead to lower failure rates than previously reported for PLC repair. Case series; Level of evidence, 4. A total of 28 patients with multiligament knee injuries who underwent acute repair between January 2007 and June 2018 of a PLC avulsion injury with no evidence of midsubstance tearing were included. All PLC avulsion injuries were treated using a transosseous Krackow suture pull-through technique without graft augmentation. Outcome metrics included lateral joint-space widening with varus stress, patient-reported clinical varus instability, patient-reported outcome measures (PROMs), and any subsequent revision or salvage procedure. The mean time from injury to repair was 8.1 ± 5 days. At a mean follow-up of 2 years (range, 3-90 months), clinical varus stress examination at 30° demonstrated a significant reduction in lateral compartment opening, from 9 ± 3 mm preoperatively to 0 ± 3 mm (P < .0001). The failure rate was calculated to be 10.7% (3/28), which was significantly lower than the failure rate from a 2016 systematic review (38%, 17/45; P = .015). Of the 28 patients, 21 (75%) had PROM scores. Patients who underwent staged bi-cruciate reconstructions (n = 5) had significantly higher subjective International Knee Documentation Committee (IKDC) (87.2 vs 65.5; P = .014) and Lysholm (90.5 vs 75.2; P = .029) scores compared to patients with untreated bi-cruciate injuries (n = 9). Patients with peroneal nerve injury (n = 4) had significantly lower IKDC (58.2 vs 80.8; P = .0045) and Tegner (3.2 vs 5.4; P = .047) scores than those without peroneal nerve injury (n = 17). The mean IKDC and Lysholm scores at final follow-up were 73.4 ± 24.0 and 80.8 ± 23.1 at 7.1 years (range, 2.3-10.6 years) of follow-up. Repair of acute grade 3 combined PLC avulsion injuries using a transosseous Krackow suture pull-through technique demonstrated a failure rate of 10.7%. Patients who underwent a staged cruciate reconstruction(s) had higher subjective outcome scores than those who had cruciate injuries left untreated. Peroneal nerve injury was associated with lower outcome scores.


Histological evaluation of colonic anastomotic healing in the rat following preoperative 5-fluorouracil, fractionated irradiation, and combined treatment.
There is a growing interest in neoadjuvant chemo- and radiotherapy as a treatment modality for colorectal cancer which could affect mechanical and biochemical parameters of anastomotic healing. This study investigated the effect of such protocols on colonic anastomotic healing by evaluating the histopathological parameters. One hundred and sixty male Wistar rats were divided into six groups: a control group (I, n = 20), a saline group (II, n = 30) which received 1 ml NaC1 intraperitoneally, a sham-irradiated group (III, n = 20), a 5-fluorouracil (5-FU) group (IV, n = 30), which received 5-FU (20 mg/kg) intraperitoneally for 5 consecutive days, an irradiated group (V, n = 40) which received fractionated irradiation to the whole pelvis to a total dose of 22 Gy, 5.5 Gy per fraction on 4 consecutive days, and a concomitant 5-FU + irradiation group (VI, n = 20) which received 5-FU as in group IV and irradiated as in group V. All groups underwent left colonic resection with primary anastomosis, and the last fraction of irradiation and the last injection were given 4 and 3 days before the operation, respectively. Within each group one half of the animals were killed on the third postoperative day and the other half on the seventh postoperative day. After the resection of the anastomotic segments, histopathological examination was evaluated. Apposition of the wound edges of the mucosa and the muscularis were not affected by the therapy. The level of granulocytes was high, inflammatory exudate and necrosis persisted, granulation tissue formation was delayed, and the levels of macrophages and fibroblasts were low. We conclude that colonic anastomotic healing can be affected by the administration of preoperative chemotherapy, irradiation, and chemoirradiation.


Human enteric caliciviruses have a unique genome structure and are distinct from the Norwalk-like viruses.
Classic human enteric caliciviruses (HuCVs) have a distinctive morphology and are primarily associated with pediatric acute gastroenteritis. Although morphologically distinct from the small round structured viruses (SRSVs), the classic HuCVs are thought to be closely related and were anticipated to have a similar genome organisation. We report the first genome sequence and molecular characterisation of a classic human enteric calicivirus associated with a case of acute vomiting and diarrhoea in an infant. The RNA genome (7266 nt) is smaller than the genome of SRSVs from the two genetic groups and has a unique arrangement of open reading frames. Further analysis of the 3' terminal 3 kb from a second unrelated isolate confirmed this genomic organisation. Analysis of capsid and RNA polymerase sequences together with the unique genomic organisation of classic HuCV suggest these viruses are more closely related to the animal caliciviruses than the enteric SRSV group of viruses.


Visual aesthetic quality of Northern Ontario's forested shorelines.
Only a few empirical studies on forest aesthetics have adopted a water-based perspective for observers and have investigated the perceived visual quality of forested shorelines. In forested environments with many lakes, such as the boreal forest in the Canadian Shield, individuals have greater exposure to forests from water-based rather than in-stand vantage points. This study employed the psychophysical research direction to explore the relationships between scenic beauty and biophysical characteristics of the forested shorelines in the boreal forests. Two model forms were tested. One model related the variation of shoreline forest aesthetic evaluations of near-vista views (140 m offshore) to a set of forest mensuration data. Tree size, tree mortality, conifer shrubs, tree density, amount of hardwood, and slope explained 60.2% of the variance in scenic beauty between the study sites. A second model was calibrated to test the relationship between an already existing ecosystem vegetation classification system and the aesthetic evaluations of the same forested shorelines. When the ecosystem classification was simplified to eight groups, the model explained 48.5% of variance. These models suggest that the psychophysical approach to studying aesthetics can be applied successfully to near-vista evaluations of scenic beauty. The finding that a forest ecosystem classification system is highly related to scenic beauty suggests that, at least in the boreal forest, managers can reasonably estimate the scenic beauty of forested shoreline environments from an ecosystem classification, with little need for intensive data on these sites.


Construction of a 5'-controllable stabilizing element (CoSE) for over-production of heterologous proteins at high levels in Bacillus subtilis.
Different mRNA stabilizing elements including the 3'-stem-loop, the ribosome binding sites (RBS), the 5'-stem-loop and the spacer region between the RBS and the 5'-stem-loop were analysed in detail to increase mRNA stability resulting in enhanced expression of heterologous proteins. In addition, in combination with mRNA stabilizing elements, we propose a new class of 5'-mRNA controllable stabilizing element (CoSE) which is composed of a transcriptional operator such as lacO of the Escherichia coli lac operon and a suitable RBS followed by an optimal spacer length. Such a CoSE allowed Bacillus subtilis cells to synthesize extraordinary stable transcripts with a half-life of the model bgaB reporter transcript (codes for an β-galactosidase gene derived from Bacillus stearothermophilus) of more than 60 min. This CoSE will be an important tool to control mRNA stability in cells for both research and biotechnological applications. For example, this CoSE can be used in inducible expression vectors offering two major advantages: (i) controlling transcription of target genes by the inducer and (ii) enhancing the stability of the transcript allowing the production high levels of recombinant proteins.


A case of oculocutaneous sarcoidosis.
To present a case of extrapulmonary sarcoidosis presenting with ocular and cutaneous involvement. We report a 54-year-male who presented with bilateral redness of eyes, photophobia, and diminished vision for a week. The best corrected visual acuity in the right eye was 6/60 and the left eye was counting fingers close to face (CFCF). He also had multiple brown plaques on the nape of the neck, chest, back, and arms. Furthermore, he was on multiple antipsychotic drugs for schizophrenia for 3 years. Uveitis investigation workup revealed raised serum angiotensin converting enzyme (ACE), negative Mantoux, and other serological tests. The patient was treated for acute anterior uveitis secondary to sarcoidosis. Clinical improvement was seen after a few days following treatment. The patient presented a year later with multiple yellowish conjunctival nodules in the superior bulbar conjunctiva associated with hyperemia. A biopsy of the plaque like skin lesions was done, which suggested cutaneous sarcoidosis. Involvement of the skin and the eyes raised suspicion that the persistent psychotic episodes despite multiple antipsychotic drugs could be attributed to neurosarcoidosis. However, magnetic Resonance Imaging (MRI) of the brain and orbit showed normal findings. After treatment with corticosteroids and immunosuppressives (methotrexate), the conjunctival nodules as well as skin lesions drastically improved, and the psychosis also responded well to clozapine. A high index of suspicion is needed in cases presenting with granulomatous uveitis with multisystem involvement. Long-term follow-up is crucial to monitor the disease progression and adverse effects of medications.


[Chronic musculoskeletal pain syndrome in primary hyperparathyroidism].
Chronic somatic pain disorders with somatic and mental factors (ICD-10: F45.41) are common among psychosomatic patients. In the present case, due to the close temporal association with a trauma and the subsequent development of symptoms including depressive symptoms, a chronic pain disorder with a relevant somatoform component was suspected. However, after a period of several months without significant somatic findings, targeted diagnostic approaches resulted in the diagnoses of primary hyperparathyroidism and a papillary thyroid carcinoma. Surgical therapy resulted in an almost complete decline of symptoms within a short period of time.


[Clinical pharmacokinetic of maraviroc].
Maraviroc (MVC, UK-427,857) is the first member of a new class, the CCR5 antagonists. By an original mechanism of action, maraviroc binds to the CCR5 receptor in order to prevent HIV from binding and entering human cells. Maraviroc (Celsentri) is an orally administered drug available as 150 and 300 mg film-coated tablets. The current approved daily dosage of maraviroc is 300 mg bid in combination with other antiretroviral medications. Maraviroc plasma exposure is not dose proportional. After a rapid (but moderate) intestinal absorption, several inactive oxidized metabolites are produced via cytochrome P450 3A4 pathway. According to this liver metabolism, dosage adjustments are required when maraviroc is administered in combination with cytochrome P450 inhibitors or inducers. The potential for drug-drug interactions and the well-defined relationship between plasma concentrations and virological response suggest the usefulness of Therapeutic Drug Monitoring of maraviroc in HIV-infected patients.


Fluorescent transgenes to study interphase chromosomes in living plants.
Fluorescence tagging of genomic sites through the use of bacterial operator/repressor systems combined with fluorescent proteins permits high-resolution analysis of interphase chromosomes in living cells. This technique has been used to study interphase chromosome arrangement and dynamics in yeast, Drosophila, and mammalian cells, but is only beginning to be exploited in plant systems. In this chapter, we describe methods for producing and identifying Arabidopsis thaliana plants harbouring fluorescence-tagged transgenes. The use of these plants to analyze various aspects of interphase chromosome organization and dynamics in living cells using 3D wide-field fluorescence microscopy is discussed. Potential problems encountered when utilizing this technology in plants are considered.


The 200-kb segmental duplication on human chromosome 21 originates from a pericentromeric dissemination involving human chromosomes 2, 18 and 13.
Regions close to human centromeres contain DNA fragments spanning hundreds of kilobases that exhibit a high degree of sequence identity (>95%). Here we report the genomic structure and evolution of a family of four paralogous regions related to a 220-kb genomic fragment present on the long arm of human chromosome 21 (21q22.1). Phylogenetic classification of the paralogous sequences obtained from the draft of the Human Genome Project are in agreement with results from comparative fluorescence in situ hybridization on metaphase chromosomes from human and great apes. The original copy present in 21q22.1 in human was duplicated in great apes after the divergence of the orang-utan and inserted in a pericentromeric region, most likely the ancestor of HSA2q, then disseminated by transposition of a larger fragment to other pericentromeric locations: HSA18p11, HSA13q11 and HSA21q11.1. The degree of dissemination varies among species.


Genotoxicity of 6 oxime compounds in the salmonella/mammalian-microsome assay and mouse lymphoma TK +/- assay.
To aid in the selection of chemical candidates for in vivo tests, the mutagenicity of 6 oxime compounds was evaluated in the Salmonella plate incorporation assay and mouse lymphoma L5178Y TK +/- assay. All of the oximes were mutagenic in the mouse lymphoma assay in the absence of exogenous metabolic activation. Acetaldehyde oxime was also mutagenic in the presence of S9 activation. In contrast to these results, a positive response was noted only for 2-(hydroxyimino)-N-phenyl-acetamide oxime in strain TA1535 in the absence of activation in the Salmonella/microsome test.


Evaluation of the reinforcing effects of the cannabinoid CB1 receptor antagonist, SR141716, in rhesus monkeys.
The abuse liability of a selective cannabinoid CB1 receptor antagonist, SR141716 (N-(piperidin-1-yl)-5-(4-chlorophenyl)-1-(2,4-dichlorophenyl)-4-methyl-1H-pyrazole-3-carboxyamide hydrochloride), was evaluated in rhesus monkeys. Four rhesus monkeys with chronically indwelling venous catheters were initially trained to self-administer cocaine (30 microg/kg/injection) during daily 1-h sessions under a fixed ratio 50 (FR50) schedule of reinforcement. SR141716 was subsequently substituted for cocaine, and SR141716 dose was varied from 1 to 100 microg/kg/injection. Each dose of SR141716 was tested for four consecutive sessions and each unit dose was separated by at least three sessions of cocaine self-administration. Substitution of SR141716 for cocaine resulted in rapid extinction of lever pressing and none of the doses of SR141716 tested was self-administered above the vehicle levels. When the highest dose of SR141716 (100 microg/kg/injection) was evaluated, self-administration behavior was suppressed below vehicle levels suggesting that behaviorally active doses were evaluated. Since positive results in self-administration tests are generally predictive of abuse potential, the negative results with SR141716 suggest that this drug would likely have low abuse liability.


Antipyrine elimination and hepatic microsomal enzyme activity in patients with liver disease.
Induction of hepatic monooxygenases reflected by 7-ethoxycoumarin O-deethylase has been proposed to be associated with the initiation of liver damage. This study investigated a possible correlation between 7-ethoxycoumarin O-deethylase, reduced nicotinamide adenine dinucleotide phosphate cytochrome c reductase and benzypyrene hydroxylase activity in liver biopsy specimens of 31 patients with liver disease and antipyrine elimination, an in vivo parameter of hepatic monooxygenase activity. No correlation was found between the enzyme activities and antipyrine clearance or half-life. When microsomal enzyme activities were compared with the formation rate of 4-hydroxyantipyrine, 3-methylhydroxyantipyrine, and norantipyrine, a correlation was found only between benzo[alpha]pyrene hydroxylase and 3-methylhydroxyantipyrine (r = 0.89; p less than 0.0005). There was also a correlation between 7-ethoxycoumarin O-deethylase and reduced nicotinamide adenine dinucleotide phosphate cytochrome c reductase (0.56; p less than 0.05). Our data suggest that antipyrine elimination is not related to 7-ethoxycoumarin O-deethylase activity in liver disease. However, the formation rate of antipyrine metabolites, rather than antipyrine half-life and clearance, may correlate with the activity of certain microsomal enzymes.


Whole slide image features predict pathologic complete response and poor clinical outcomes in triple-negative breast cancer.
Breast cancers are complex ecosystem like networks of malignant cells and their associated microenvironment. Applications for machine intelligence and the tumoral microenvironment are expanding frontiers in pathology. Previously, computational approaches have been developed to quantify and spatially analyze immune cells, proportionate stroma, and detect tumor budding. Little work has been done to analyze different types of tumor-associated stromata both quantitatively and computationally in relation to clinical endpoints. We aimed to quantify stromal features from whole slide images (WSI) including stromata (myxoid, collagenous, immune) and tumoral components and combined them with traditional clinical and pathologic parameters in 120 triple-negative breast cancer (TNBC) patients treated with neoadjuvant chemotherapy (NAC) to predict pathologic complete response (pCR) and poor clinical outcomes. High collagenous stroma on WSI was best associated with lower rates of pCR, while combined high proportionated stroma (myxoid, collagenous, and immune) most optimally predicted worse clinical survival outcomes. When combining clinical, pathologic, and WSI features, Receiver Operator Characteristics (ROC) curves for LASSO features was up to 0.67 for pCR and 0.77 for poor outcomes. The techniques demonstrated in the present study can be performed with appropriate quality assurance. Future trials are needed to demonstrate whether coupling applications for machine intelligence, inclusive of the tumor mesenchyme, can improve outcomes prediction for patients with breast cancer.


Valsaria and the Valsariales.
More than 100 recent collections of Valsaria sensu lato mostly from Europe were used to elucidate the species composition within the genus. Multigene phylogeny based on SSU, LSU, ITS, rpb2 and tef1 sequences revealed a monophyletic group of ten species within the Dothideomycetes, belonging to three morphologically similar genera. This group could not be accommodated in any known family and are thus classified in the new family Valsariaceae and the new order Valsariales. The genus Valsaria sensu stricto comprises V. insitiva, V. robiniae, V. rudis, V. spartii, V. lopadostomoides sp. nov. and V. neotropica sp. nov., which are phylogenetically well-defined, but morphologically nearly indistinguishable species. The new monotypic genus Bambusaria is introduced to accommodate Valsaria bambusae. Munkovalsaria rubra and Valsaria fulvopruinata are combined in Myrmaecium, a genus traditionally treated as a synonym of Valsaria, which comprises three species, with M. rubricosum as its generic type. This work is presented as a basis for additional species to be detected in future.