Does an education intervention improve physician signature legibility? Pilot study of a prospective chart review.
Illegible physician signatures in patient records can lead to inaccurate documentation, improper billing, and potential legal issues. Many studies in the current literature address legibility of prescriptions and medication orders; however, few focus on legibility of physicians' signatures. The purpose of the present quality improvement survey was to evaluate physician signature legibility on patient charts at the University of Mississippi Medical Center's Adult Internal Medicine Clinic. At the time of the study, the clinic was known as the University of Mississippi Medical Center (UMMC) Adult Internal Medicine Clinic. Effective July 1, 2009, UMMC entered into a collaboration with Jackson-Hinds Comprehensive Health Center (JHCHC), a federally qualified health center. The clinic is now known as the Federally Qualified Health Center at the Jackson Medical Mall. In this pilot study, we examined clinic notes and billing sheets for legible physician signatures over a three-month period. Midway through the study, an intervention group was given name stamps and a standardized discussion on the importance of signature legibility and proper name stamp usage. Legibility of resident signatures in the intervention group increased from 26 percent to 60 percent. Legibility of attending signatures in the intervention group increased from 1.4 percent to 86 percent. Results suggest the significant impact of resident education on changing practice behavior.


Effects of mulberry twig alkaloids(Sangzhi alkaloids) and metformin on blood glucose fluctuations in combination with premixed insulin-treated patients with type 2 diabetes.
We aimed to evaluated the effect of premixed insulin (Ins), premixed insulin combined with metformin (Ins+Met) or mulberry twig alkaloids(Ins+SZ-A) on blood glucose fluctuations in patients with type 2 diabetes (T2DM) using continuous glucose monitors (CGM). Thirty patients with T2DM and poor blood glucose control using drugs were evaluated for eligibility during the screening period. Subsequently, their original hypoglycemic drugs were discontinued during the lead-in period, and after receiving Ins intensive treatment for 2 weeks, they were randomly assigned to receive either Ins, Ins+Met, or Ins+SZ-A treatment for the following 12 weeks. The main efficacy endpoint comprised changes in their CGM indicators changes (mean blood glucose level [MBG], standard deviation of blood glucose [SDBG], mean amplitude of glycemic excursions [MAGE], postprandial glucose excursions [PPGE], the largest amplitude of glycemic excursions [LAGE], mean of daily difference [MODD], time in range between 3.9-10.0 mmol/L [TIR] and area under the curve for each meal [AUCpp]) during the screening, lead-in, and after 12-week treatment period. Changes in glycosylated hemoglobin (HbA1c), fasting blood glucose (FBG), 1-h postprandial blood glucose (1h-PBG), 2-h postprandial blood glucose (2h-PBG), fasting blood lipids and postprandial blood lipids were also measured at baseline and after 12 weeks of treatment. The CGM indicators of the three groups during the lead-in period all showed significant improvements compared to the screening period (P<0.05). Compared with those in the lead-in period, all of the CGM indicators improved in the the Ins+Met and Ins+SZ-A groups after 12 weeks of treatment (P<0.05), except for MODD. After 12-week treatment, compared with the Ins group, Ins+Met and Ins+SZ-A groups showed improved MBG, SDBG, TIR, breakfast AUCpp,lunch AUCpp, HbA1c, FBG, 1h-PBG, fasting blood lipid and postprandial blood lipid indicators (P<0.05). Further, the LAGE, PPGE, MAGE, dinner AUCpp and 2h-PBG levels of the Ins+SZ-A group were significantly lower than those of the Ins+Met and Ins groups (P<0.05). Our findings highlight the efficacy of combination therapy (Ins+SZ-A or Ins+Met) in improving blood glucose fluctuations, as well as blood glucose and lipid levels. Ins+SZ-A reduces postprandial blood glucose fluctuations more than Ins+Met and Ins groups. ISRCTN20835488.


Photoreceptor rescue by an abbreviated human RPGR gene in a murine model of X-linked retinitis pigmentosa.
The X-linked RP3 gene codes for the ciliary protein RPGR and accounts for over 10% of inherited retinal degenerations. The critical RPGR-ORF15 splice variant contains a highly repetitive purine-rich linker region that renders it unstable and difficult to adapt for gene therapy. To test the hypothesis that the precise length of the linker region is not critical for function, we evaluated whether adeno-associated virus-mediated replacement gene therapy with a human ORF15 variant containing in-frame shortening of the linker region could reconstitute RPGR function in vivo. We delivered human RPGR-ORF15 replacement genes with deletion of most (314 codons, 'short form') or 1/3 (126 codons, 'long form') of the linker region to Rpgr null mice. Human RPGR-ORF15 expression was detected post treatment with both forms of ORF15 transgenes. However, only the long form correctly localized to the connecting cilia and led to significant functional and morphological rescue of rods and cones. Thus the highly repetitive region of RPGR is functionally important but that moderate shortening of its length, which confers the advantage of added stability, preserves its function. These findings provide a theoretical basis for optimizing replacement gene design in clinical trials for X-linked RP3.


Cadmium-113 magnetic resonance spectroscopy.
Cadmium-113 nuclear magnetic resonance spectroscopy has been used in studies of the structure and dynamics of inorganic and bioinorganic molecules. Chemical dynamics play an important role in the analysis of relaxation and chemical shift data. Naïve interpretations of relaxation data can be checked by performing these experiments at a variety of temperatures and magnetic field strengths. A combination of solid- and liquid-state nuclear magnetic resonance measurements can provide the user with unambiguous data on chemical shielding. These data can be used to characterize zinc and calcium ion binding sites in metalloproteins.


Tuning and Probing the Distribution of Cu+ and Cu2+ Trap States Responsible for Broad-Band Photoluminescence in CuInS2 Nanocrystals.
The processes that govern radiative recombination in ternary CuInS2 (CIS) nanocrystals (NCs) have been heavily debated, but recently, several research groups have come to the same conclusion that a photoexcited electron recombines with a localized hole on a Cu-related trap state. Furthermore, it has been observed that single CIS NCs display narrower photoluminescence (PL) line widths than the ensemble, which led to the conclusion that within the ensemble there is a distribution of Cu-related trap states responsible for PL. In this work, we probe this trap-state distribution with in situ photoluminescence spectroelectrochemistry. We find that Cu2+ states result in individual "dark" nanocrystals, whereas Cu+ states result in "bright" NCs. Furthermore, we show that we can tune the PL position, intensity, and line width in a cyclic fashion by injecting or removing electrons from the trap-state distribution, thereby converting a subset of "dark" Cu2+ containing NCs into "bright" Cu+ containing NCs and vice versa. The electrochemical injection of electrons results in brightening, broadening, and a red shift of the PL, in line with the activation of a broad distribution of "dark" NCs (Cu2+ states) into "bright" NCs (Cu+ states) and a rise of the Fermi level within the ensemble trap-state distribution. The opposite trend is observed for electrochemical oxidation of Cu+ states into Cu2+. Our work shows that there is a direct correlation between the line width of the ensemble Cu+/Cu2+ trap-state distribution and the characteristic broad-band PL feature of CIS NCs and between Cu2+ cations in the photoexcited state (bright) and in the electrochemically oxidized ground state (dark).


Enhancing Collaborative Neuroimaging Research: Introducing COINSTAC Vaults for Federated Analysis and Reproducibility.
Collaborative neuroimaging research is often hindered by technological, policy, administrative, and methodological barriers, despite the abundance of available data. COINSTAC is a platform that successfully tackles these challenges through federated analysis, allowing researchers to analyze datasets without publicly sharing their data. This paper presents a significant enhancement to the COINSTAC platform: COINSTAC Vaults (CVs). CVs are designed to further reduce barriers by hosting standardized, persistent, and highly-available datasets, while seamlessly integrating with COINSTAC's federated analysis capabilities. CVs offer a user-friendly interface for self-service analysis, streamlining collaboration and eliminating the need for manual coordination with data owners. Importantly, CVs can also be used in conjunction with open data as well, by simply creating a CV hosting the open data one would like to include in the analysis, thus filling an important gap in the data sharing ecosystem. We demonstrate the impact of CVs through several functional and structural neuroimaging studies utilizing federated analysis showcasing their potential to improve the reproducibility of research and increase sample sizes in neuroimaging studies.


Disinfection of Stethoscope and Non-Infrared Thermometer: Practices of Physicians in Ethiopia in the Era of COVID-19.
Stethoscopes and non-infrared thermometers are the customary medical equipment used by the physicians on a daily basis, among various patients. With the rise of potential infections in the healthcare facilities and the transmission nature of the current COVID-19 pandemic, consistent and correct disinfections of these devices after each use should not be pardoned. This study, therefore, aimed to assess the level of stethoscope and non-infrared thermometer disinfection practices among physicians involved in direct patient contact during the COVID-19 pandemic. A web-based cross-sectional survey was conducted among physicians working in Ethiopia to assess their practice of stethoscope and non-infrared thermometer disinfection. The online survey was circulated using an anonymous and self-reporting questionnaire via Google form with a consent form appended to it. The developed Google form link was shared with physicians through their email addresses and social media pages. A descriptive summary was computed and presented by tables and figures. Multivariable logistic regression model was used to identify factors associated stethoscope and non-infrared thermometer after every use. The proportion of stethoscope and non-infrared thermometer disinfections after every use was 13.9% (95% CI: 10.9-17.6) and 20.4% (95% CI: 16.7-24.5), respectively. Taking COVID-19 training (AOR: 2.52; 95% CI: 1.29-4.92) and the availability of stethoscope disinfection materials at the workplace (AOR: 3.03; 95% CI: 1.29-7.10) were significantly increased the odds of stethoscope disinfection after every use. The odds of stethoscope disinfection after every use was significantly decreased for those who reported the use of shared stethoscope (AOR: 0.34; 95% CI: 0.12-0.92). Only a wee share of the respondents reported that they have disinfected their stethoscopes and non-infrared thermometers after every use - possibly jeopardizing both patients and clinicians safety, particularly during the COVID-19 pandemic.


Mental Health and Cognition in Older Cannabis Users: a Review.
The impact of cannabis use on mental health and cognition in older adults remains unclear. With the recent legalization of cannabis in Canada, physicians will need up-to-date information about the mental and cognitive effects of cannabis use in this specific population. A narrative review was conducted to summarize the literature on mental health and cognitive effects of cannabis use in older adults using Medline (OvidSP). A total of 16 studies were identified, including nine cross-sectional studies on mental health comorbidities reported by older cannabis users. The self-reported prevalence of mental and substance use disorders is approximately two to three times higher in older adults who report past-year cannabis use, compared to older adults who report using more than one year ago or never using. The remaining seven clinical trials found that short-term, low-dose medical cannabis was generally well-tolerated in older adults without prior serious mental illness. However, mental/cognitive adverse effects were not systematically assessed. Although preliminary findings suggests that low-dose, short-term medical cannabis does not carry significant risk of serious mental health and cognitive adverse effects in older adults without prior psychiatric history, epidemiological studies find a correlation between past-year cannabis use and poor mental health outcomes in community-dwelling older adults. These findings may indicate that longer term cannabis use in this population is detrimental to their mental health, although a direct causal link has not been established. Larger, longitudinal studies on the safety of medical cannabis in older adults are needed.


A possible physiological basis for effectiveness of acupuncture against psychosomatic disorders.
Acupuncture and Oriental meditation, particularly of Zen Buddhists have essential features in common. The physiological correlates of these features are discussed as they apply to therapy of psychosomatic disorders. Both acupuncture and meditation are characterized by alpha rhythms of the encephalograms, by relaxed wakefulness, by analgesia, by their pervasion of virtually the entire body. This set of correspondences is taken as sufficient to postulate that acupuncture induces a stillness with properties similar to those of the "quiet" of meditation. This induced deep relaxation may account for the vasolidation and increased vascular-capillary flow, intrinsic in acupuncture. Among the shared properties are the presence of low physiological noise-level and unresponsiveness to the aversive components of conditioned stimuli.


An overview of trials of cholesterol lowering and risk of stroke.
While blood cholesterol level predicts coronary heart disease, whether there is any association with the risk of stroke is unclear. Some, but not all, observational studies suggest that cholesterol level predicts risk of stroke, particularly ischemic stroke. This hypothesis is attractive because ischemic events constitute the vast majority of all strokes and, like coronary heart disease, involve atherogenic processes. To investigate whether lipid lowering reduces the risk of stroke, we performed an overview of randomized trials that included more than 36,000 individuals. The mean reduction in cholesterol level in the treated as compared with the control subjects ranged from 6% to 23%. Those assigned to treatment experienced no significant reduction in all (fatal plus nonfatal) stroke (relative risk, 1.0; 95% confidence interval, 0.8 to 1.2) or fatal stroke (1.1; 0.8 to 1.6). The confidence interval for fatal stroke is wide, and alternative hypotheses, including either a small protective or harmful effect, cannot be excluded; however, the point estimates are compatible with no benefit of cholesterol lowering on the risk of stroke. Additional large-scale randomized trials assessing total mortality would more definitively address any benefits on stroke, as well as any excess nonvascular causes of mortality, for which risks of cholesterol lowering also remain uncertain.


Influence of psychological symptoms on home-recorded sleep-time masticatory muscle activity in healthy subjects.
The present investigation attempts to describe the correlation between sleep-time masticatory muscle activity (MMA) and psychological symptoms by the use of a four-channel electromyography (EMG) home-recording device in a group of 15 healthy volunteers completing a battery of psychometric questionnaires for the assessment of anxiety, depression and anger. The integrated EMG signal was adopted to quantify the work (μV × s) produced by each of the four muscles (bilateral masseter and temporal) during the 5-h recording span and per each 1-h increment. The duration of MMA events and the muscle work during the first hour of sleep was related to trait anxiety scores for both masseter (P = 0·007) and temporalis muscles (P = 0·022). Trait anxiety was also significantly correlated to the total amount of MMA duration (in seconds) of the temporalis muscles (r = 0·558; P = 0·031). The present investigation provides support to the hypothesis that the duration of sleep-time masticatory muscle activity, especially during the early phases of a night's sleep, may be related to anxiety trait and not to anxiety state, depression or anger. These findings may support the view that features related to the individual management of anxiety, viz. trait, are likely to be more important than acute episodes of anxiety, viz. state, in the aetiology of sleep-time masticatory muscle activity. The role of other psychological symptoms is likely to be less important.


Treatment of Pulp Black Liquor with an Ionic Liquid: Modeling and Optimization by Using Response Surface Methodology and Central Composite Design.
In this study, selective extraction towards ionic liquid trihexyltetradecylphosphonium chloride ([P66614 ]Cl) of lignin residues, polysaccharides and organic acids present in black liquor (BL), the main principal wastewater of pulping industry was studied. With the objective of finding optimized conditions allowing to extract lignin residues while polysaccharides and organic acids remain in aqueous solution, a design of experiments approach based on a response surface methodology was used. Three continuous factors, namely initial pH varying from 9 to 13.5, dilution of BL varying from 5 to 20 and volumetric ratio of black liquor vs. ionic liquid RV ., varying from 1 to 19, were investigated. Concentration of lignin residues, polysaccharides and organic acids were measured using Folin-Ciocalteu method, the anthrone method and HPLC, respectively. Results showed that a multi-response optimization led to the extraction of 84.8 % of lignin residues, 66.0 % of polysaccharides, and no extraction of OA under optimised conditions.


Co-occurring implementation strategies: The effects of academic detailing for opioid use disorder campaign on the advancing pharmacological treatments for opioid use disorder (ADaPT-OUD) study.
Barriers at the system, clinician, and patient level limit access to medications for opioid use disorder (MOUD). The Advancing Pharmacological Treatments for Opioid Use Disorder (ADaPT-OUD) study implemented an external facilitation strategy within the Veterans Health Administration (VHA) aimed at facility-level barriers to improve uptake of MOUD. During ADaPT-OUD, an independent Academic Detailing Services Opioid Agonist Treatment of OUD Campaign was co-occurring and aimed to increase evidence-based practice for OUD at the clinician level. While both these initiatives aim to increase MOUD reach, they address different barriers and did not intentionally collaborate. Thus, understanding the interaction between these two independent implementation initiatives and their effect on MOUD reach will further inform and mold future implementation efforts of MOUD. This was a secondary analysis of the ADaPT-OUD study that included 35 VHA facilities in the lowest quartile of MOUD reach; eight received the ADaPT-OUD external facilitation and 27 matched sites received implementation as usual. The number of academic detailing (AD) visits during ADaPT-OUD was used as a proxy for the intensity of Academic Detailing for OUD Campaign activity. The interaction between external facilitation status and AD intensity was evaluated by comparing the change in facility-level MOUD reach. There was a general increase in the number of AD visits, in both external facilitation and implementation as usual sites, over the course of ADaPT-OUD's implementation period. A non-statistically significant, positively sloped, linear relationship was observed between average number of AD visits per quarter and change in MOUD reach in facilities also receiving ADaPT-OUD external facilitation that was not observed in the implementation as usual sites. Co-occurring initiatives focusing on different barriers to MOUD access have the potential to further increase MOUD in low-performing facilities, but further research into timing, quality, and collaboration between initiatives are warranted.


Endoscopic third ventriculostomy for idiopathic aqueductal stenosis.
Idiopathic aqueductal stenosis is a cause of noncommunicating hydrocephalus, which actual treatment with endoscopic third ventriculostomy (ETV) could assess without any interference with the etiology. The results of ETV in this indication therefore could be interpreted as the result of the surgical procedure alone, without any additional factors related to the etiology of the cerebrospinal fluid pathway obstruction, such as hemorrhage, infection, brain malformations, or brain tumors or cysts. After a brief description of pathogenesis of hydrocephalus in aqueductal stenosis, the authors review the literature for studies on ETV, extrapolating patients with idiopathic aqueductal stenosis in infancy, childhood, and adulthood. Differences in outcome between patients treated with ETV and patients treated with ventriculoperitoneal shunt (VPS) are also reviewed. The overall success rates of ETV range between 23% to 94%, with a mean of 68%; when only patients affected by obstructive triventricular hydrocephalus secondary to aqueductal stenosis are considered, the success rate is actually quite homogeneous and stable, being above 60% at any age, even if a trend in lower success rate in very young infants (younger than 6 months of age) is noticeable. The few reports on intellectual outcome failed to demonstrate differences between ETV and VPS. Several issues, such as the cause of failures in well-selected patients, long-term outcome in infants treated with ETV, effects of persistent ventriculomegaly on neuropsychological developmental, remain unanswered. Larger and more detailed studies are needed.


Value of bone scan in primary sternal osteomyelitis.
Roentgenographic examination of the sternum is particularly difficult and offers little clinical assistance in the early stage of acute osteomyelitis. With only a few cases of primary sternal osteomyelitis reported, there are almost no data available regarding the role of bone scan in the diagnosis of this lesion. The present case illustrates the usefulness of the scan both in diagnosis and management of sternal osteomyelitis.


Cognitive bias in forensic pathology decisions.
Forensic pathologists' decisions are critical in police investigations and court proceedings as they determine whether an unnatural death of a young child was an accident or homicide. Does cognitive bias affect forensic pathologists' decision-making? To address this question, we examined all death certificates issued during a 10-year period in the State of Nevada in the United States for children under the age of six. We also conducted an experiment with 133 forensic pathologists in which we tested whether knowledge of irrelevant non-medical information that should have no bearing on forensic pathologists' decisions influenced their manner of death determinations. The dataset of death certificates indicated that forensic pathologists were more likely to rule "homicide" rather than "accident" for deaths of Black children relative to White children. This may arise because the base-rate expectation creates an a priori cognitive bias to rule that Black children died as a result of homicide, which then perpetuates itself. Corroborating this explanation, the experimental data with the 133 forensic pathologists exhibited biased decisions when given identical medical information but different irrelevant non-medical information about the race of the child and who was the caregiver who brought them to the hospital. These findings together demonstrate how extraneous information can result in cognitive bias in forensic pathology decision-making.


Prostacyclin production by perturbed bovine aortic endothelial cells in culture.
This study reports that endotoxin (Escherichia coli serotype 026:B6) and 12-O-tetradecanoyl-phorbol-13-acetate stimulate cultured bovine aortic endothelial cells to generate prostacyclin. The prostacyclin concentration of the culture medium was measured indirectly by radioimmunoassay for 6-keto-PGF1 alpha. The amount of prostacyclin generated depended on the concentration of endotoxin or phorbol diester. Prostacyclin generation was not immediate, but occurred slowly after a six-hour lag period. The perturbed cells contracted and showed marked shape changes that correlated temporally with the start of enhanced prostacyclin production. Cytochalasins B and D, vinblastine, and colchicine inhibited prostacyclin production, indicating involvement of the cytoskeleton in the cellular response to endotoxin and phorbol diester. The increase in prostacyclin production was prevented by trifluoperazine, an inhibitor of the Ca++-calmodulin system, which is known to be involved in cytoskeletal function. Generation of prostacyclin was inhibited by cycloheximide and actinomycin D, indicating dependence on protein and ribonucleic acid synthesis. It is postulated that exposure to endotoxin or phorbol diester leads, via a series of reactions that involve RNA and protein synthesis and require intact cytoskeletal function, to the generation of toxic active intermediate(s) that stimulate the enzymes necessary for prostacyclin production.


Elevated inflammation in association with alcohol abuse among Blacks but not Whites: results from the MIDUS biomarker study.
Some studies document racial disparities in self-reported health associated with alcohol use and abuse. However, few studies examined biomarkers that underlie the onset of alcohol-related chronic diseases. We investigated whether the association between alcohol abuse and five biomarkers of inflammation (CRP, IL-6, fibrinogen, E-selectin, sICAM-1) vary between Black and White Americans aged 35 to 84 (n = 1173) from the Midlife in the United States Biomarker Study. Multivariable Ordinary Least Squares regressions were used to assess Black-White differences in the association between alcohol abuse and the biomarkers. Race moderated the association between alcohol abuse and CRP (b = 0.56, SE = 0.28, p = 0.048), IL-6 (b = 0.65, SE = 0.22, p = 0.004), and a composite inflammation score (b = 0.014, SE = 0.07, p = 0.041). These findings potentially shed light for why alcohol has a stronger negative association with poorer health for Blacks compared to Whites. Analysis should be replicated in larger prospective cohorts.


Intercalation and stitching of graphite oxide with diaminoalkanes.
The intercalation reaction of graphite oxide with diaminoalkanes, with the general formula H2N(CH2)nNH2 (n = 4-10), was studied as a method for synthesizing pillared graphite with tailored interlayer spacing. Interlayer spacings from 0.8 to 1.0 nm were tailored by varying the size of the intercalant from (CH2)4 to (CH2)10. X-ray diffraction and infrared spectroscopy were used to confirm intercalation, and the frequency of the CH2 stretch confirmed that the intercalants are in a disordered state, with an important contribution from the gauche conformer. Sequential intercalation of diaminoalkanes followed by dodecylamine demonstrated the inability of these "stitched" systems to undergo expansion along the c-direction, indicative of cross-linking. Finally, the reaction of graphite oxide with diaminoalkanes under reflux and for extended periods (>72 h) resulted in the chemical reduction of the graphite oxide to a disordered graphitic structure.


Direct-to-Consumer Genetic Testing: Helping Patients Make Informed Choices
Using direct-to-consumer genetic testing (DTCGT), individuals can order a genetic test, collect and submit a saliva sample, and obtain results about their genetic risk for a variety of traits and health conditions without involving a healthcare provider. Potential benefits of DTCGT include personal control over genetic information and health management decisions, whereas potential risks include misinterpretation of results, psychosocial distress, and lack of informed consent. Oncology nurses can provide education, support, and advocacy to enable patients to truly understand the positives and negatives associated with DTCGT. .


Adult age differences in memory: effects of distinctive and common encodings.
To clarify the role of encoding distinctiveness and encoding cue utilization in age-related memory differences, young and elderly adults were instructed either to generate distinctive or common adjectives for 40 nouns and given 3 study-recall trials for the nouns, both with no cues and with the adjectives that they had generated as cues. Their retention was compared with that of a control group that had rated the nouns for abstractness. Elderly adults were as likely as young adults to generate distinctive adjectives, but were less likely than young adults to generate common adjectives when instructed to do so. In both age groups, common adjective encodings produced superior free recall and distinctive adjective encodings produced superior cued recall. The results suggest that (1) elderly adults are as capable as young adults of generating distinctive encoding context cues when instructed to do so, and (2) age-related encoding differences occur in the processing of distinctive properties of the stimulus items themselves rather than in the utilization of cues generated during study.


Hepatocellular carcinoma in South America: Evaluation of risk factors, demographics and therapy.
Hepatocellular carcinoma (HCC) is the second leading cause of cancer-related death worldwide. Most studies addressing the epidemiology of HCC originate from developed countries. This study reports the preliminary findings of a multinational approach to characterize HCC in South America. We evaluated 1336 HCC patients seen at 14 centres in six South American countries using a retrospective study design with participating centres completing a template chart of patient characteristics. The diagnosis of HCC was made radiographically or histologically for all cases according to institutional standards. Methodology of surveillance for each centre was following AASLD or EASL recommendations. Sixty-eight percent of individuals were male with a median age of 64 years at time of diagnosis. The most common risk factor for HCC was hepatitis C infection (HCV, 48%), followed by alcoholic cirrhosis (22%), Hepatitis B infection (HBV, 14%) and NAFLD (9%). We found that among individuals with HBV-related HCC, 38% were diagnosed before age 50. The most commonly provided therapy was transarterial chemoembolization (35% of HCCs) with few individuals being considered for liver transplant (<20%). Only 47% of HCCs were diagnosed during surveillance, and there was no difference in age of diagnosis between those diagnosed incidentally vs by surveillance. Nonetheless, being diagnosed during surveillance was associated with improved overall survival (P = .01). Our study represents the largest cohort to date reporting characteristics and outcomes of HCC across South America. We found an important number of HCCs diagnosed outside of surveillance programmes, with associated increased mortality in those patients.


Performing Accurate Standard 12 Lead ECGs on patients with Burns to the Chest.
The use of the electrocardiogram (ECG) in critical care settings is a long-established cardiovascular monitoring tool. The effectiveness of the routine 12-lead ECG relies on accurate lead placement that is consistent and replicable. Improper lead placement may display erroneous ECG patterns and affect patient management decisions.1,2 In the setting of an acute injury, such as a torso burn to the ventral surface, accurate lead placement may be compromised or impossible. The regional burn center, which is part of our organization, sees approximately 500 patients per year. Of those patients, burns to the chest accounted for 21% of admissions during 2020 and 2021. This significant fraction of burn injury patients requires modification of our standard approach to provide an accurate ECG. Baseline ECGs are routinely acquired on the burn unit per protocol and for monitoring of patient response to numerous pharmaceutical therapies.


pH-responsive vesicles based on a hydrolytically self-cross-linkable copolymer.
A new type of shape-persistent, pH-responsive vesicle was prepared by the self-assembly of a novel poly(ethylene oxide)-block-poly[2-(diethylamino)ethyl methacrylate-stat-3-(trimethoxysilyl)propyl methacrylate], PEO-b-P(DEA-stat-TMSPMA), copolymer. Vesicles were formed spontaneously in aqueous THF solution, with the hydrophilic PEO chains forming the corona and the pH-sensitive P(DEA-stat-TMSPMA) blocks being located in the membrane walls. Hydrolytic cross-linking within the hydrophobic membrane walls fixed the vesicle morphology. The resulting colloidally stable vesicles were characterized by 1H NMR, transmission electron microscopy (TEM), dynamic laser light scattering (DLS), and stopped-flow fluorescence experiments. The latter technique indicated that the permeability of the vesicle walls was sensitive to the pH of the aqueous solution, as expected. Gold-decorated vesicles were obtained by in situ reduction of AuCl4- anions to produce gold nanoparticles within the vesicle walls. (Yellow, hydrophilic PEO; green, pH-responsive DEA residues; blue, hydrolytically self-cross-linkable TMSPMA residues.)


The chromosomal gene structure and two mRNAs for human granulocyte colony-stimulating factor.
Two different cDNAs for human granulocyte colony-stimulating factor (G-CSF) were isolated from a cDNA library constructed with mRNA prepared from human squamous carcinoma cells, which produce G-CSF constitutively. The nucleotide sequence analysis of both cDNAs indicated that two polypeptides coded by these cDNAs are different at one position where three amino acids are deleted/inserted. When the two cDNAs were introduced into monkey COS cells under the SV40 early promoter, both of them produced proteins having authentic G-CSF activity and some difference in the specific activity was suggested. A human gene library was then screened with the G-CSF cDNA and the DNA fragment containing the G-CSF chromosomal gene was characterized by the nucleotide sequence analysis. The human G-CSF gene is interrupted by four introns and a comparison of the structures of the two G-CSF cDNAs with that of the chromosomal gene indicated that the two mRNAs are generated by alternative use of two 5' splice donor sequences in the second intron of the G-CSF gene. When the G-CSF chromosomal gene was expressed in monkey COS cells by using the SV40 enhancer two mRNAs were detected by S1 mapping analysis.


Immunoscore Signature Predicts Postoperative Survival and Adjuvant Chemotherapeutic Benefits in Esophageal Squamous Cell Carcinoma.
The aim of this study was to construct the immunoscore (IS) to facilitate the prediction of postoperative survival and benefit from adjuvant chemotherapy (ACT) in esophageal squamous cell carcinoma (ESCC). A total of 249 patients who received radical esophagectomy at Fudan University Shanghai Cancer Center were divided into training set and testing set. Eighty-nine patients with ESCC from TCGA database were enrolled into the validation set. Myeloid cells in tumor microenvironment were evaluated by immunohistochemistry or CIBERSORT, and then were included into a LASSO Cox regression model to construct the immunoscore. The predictive value of the immunoscore for prognosis after surgery or ACT was analyzed. The immunoscore was constructed by four types of myeloid cells including macrophages, neutrophils, mast cells, and dendritic cells and was demonstrated as IS=2^(0.527719*Mφ -0.2604269*MC-0.4812935*DC-0.4519706*Neu). The overall survival was significantly different between two immunotypes, which were divided according to the immunoscore, in all sets (P<0.001, P=0.005, and P=0.002, respectively). Immunotype A was identified as an independent predictor for survival benefit in all three sets (HR=2.068, P=0.005; HR=2.028, P=0.007; HR=6.474, P=0.007; respectively). In patients who received ACT, immunotype A was significantly related to longer overall survival both in the training set (P<0.001) and in the testing set (P=0.011). The nomogram based on immunotype and other clinicopathological factors showed good efficiency of predicting response to ACT. Finally, several important cytokines and pathways were highly enriched in immunoscore A subgroup. The immunoscore was an effective prognostic predictor in ESCC for patients undergoing surgical resection and receiving ACT.


Evaluation of the radial immunodiffusion enzyme assay for the detection of antibodies to pseudorabies virus.
The validity of radial immunodiffusion enzyme assay (RIDEA) as a diagnostic test for antibodies to pseudorabies virus (PRV) in porcine serum was determined. Serum samples from sows and offspring were tested for the presence of antibodies to PRV, using both the RIDEA and the PRV serum-neutralization (SN) test. Overall sensitivity and specificity of the RIDEA done on serums from the sows were 95.7% and 95.2%, respectively. This sensitivity compares with 97.3% sensitivity of the SN test of the same serums. In 658 swine serum samples from routine submissions to the University of Missouri-Columbia Veterinary Diagnostic Laboratory that were tested by the RIDEA, the calculated sensitivity and the specificity were 94.3% and 98.9%. The RIDEA and SN test were equally sensitive (99.0%) to detect antibodies resulting from infection with a field strain of virus. They had reduced sensitivity (RIDEA, 91.7%; SN test, 95.2%) in tests of serums from vaccinated sows. For the detection of passively transferred antibodies in young pigs, sensitivity of the RIDEA was 76.1%, and specificity was 100%. In all instances, RIDEA was 100% sensitive at SN titers of 1:16 or greater. In testing serum samples of swine after field virus infection, sensitivity and specificity of the RIDEA approximated those of the SN test. This reliability, together with its ease of performance, makes the RIDEA an ideal field test in programs to detect PRV-infected herds and in programs designed to free herds of PRV infection.


Results of diagnostic accuracy studies are not always validated.
Internal validation of a diagnostic test estimates the degree of random error, using the original data of a diagnostic accuracy study. External validation requires a new study in an independent but similar population. Here we describe whether diagnostic research is validated, which technique is used, and to what extent the validation study results differ from the original. All original diagnostic accuracy studies published in 1993 in a predefined set of journals were selected. Validation of these studies was assessed in the original article and in articles published within a period of 10 years, through a literature search and contacting the authors. None of the original studies reported any form of validation. Validation studies published later could be identified for 7 of the 11 original studies. Test characteristics were difficult to compare. Despite what was generally believed, not every validation study showed results inferior to the original. We found more studies that evaluated the test in a different population than in a similar one. Not every diagnostic accuracy study is validated. Diagnostic tests are more often repeated in different populations.


Electrophysiologic Characterization of Developing Human Embryonic Stem Cell-Derived Photoreceptor Precursors.
Photoreceptor precursor cells (PRPs) differentiated from human embryonic stem cells can serve as a source for cell replacement therapy aimed at vision restoration in patients suffering from degenerative diseases of the outer retina, such as retinitis pigmentosa and AMD. In this work, we studied the electrophysiologic maturation of PRPs throughout the differentiation process. Human embryonic stem cells were differentiated into PRPs and whole-cell recordings were performed for electrophysiologic characterization at days 0, 30, 60, and 90 along with quantitative PCR analysis to characterize the expression level of various ion channels, which shape the electrophysiologic response. Finally, to characterize the electrically induced calcium currents, we employed calcium imaging (rhod4) to visualize intracellular calcium dynamics in response to electrical activation. Our results revealed an early and steady presence (approximately 100% of responsive cells) of the delayed potassium rectifier current. In contrast, the percentage of cells exhibiting voltage-gated sodium currents increased with maturation (from 0% to almost 90% of responsive cells at 90 days). Moreover, calcium imaging revealed the presence of voltage-gated calcium currents, which play a major role in vision formation. These results were further supported by quantitative PCR analysis, which revealed a significant and continuous (3- to 50-fold) increase in the expression of various voltage-gated channels concomitantly with the increase in the expression of the photoreceptor marker CRX. These results can shed light on the electrophysiologic maturation of neurons in general and PRP in particular and can form the basis for devising and optimizing cell replacement-based vision restoration strategies.


Mesencephalic astrocyte-derived neurotrophic factor protects the heart from ischemic damage and is selectively secreted upon sarco/endoplasmic reticulum calcium depletion.
The endoplasmic reticulum (ER) stress protein mesencephalic astrocyte-derived neurotrophic factor (MANF) has been reported to protect cells from stress-induced cell death before and after its secretion; however, the conditions under which it is secreted are not known. Accordingly, we examined the mechanism of MANF release from cultured ventricular myocytes and HeLa cells, both of which secrete proteins via the constitutive pathway. Although the secretion of proteins via the constitutive pathway is not known to increase upon changes in intracellular calcium, MANF secretion was increased within 30 min of treating cells with compounds that deplete sarcoplasmic reticulum (SR)/ER calcium. In contrast, secretion of atrial natriuretic factor from ventricular myocytes was not increased by SR/ER calcium depletion, suggesting that not all secreted proteins exhibit the same characteristics as MANF. We postulated that SR/ER calcium depletion triggered MANF secretion by decreasing its retention. Consistent with this were co-immunoprecipitation and live cell, zero distance, photo affinity cross-linking, demonstrating that, in part, MANF was retained in the SR/ER via its calcium-dependent interaction with the SR/ER-resident protein, GRP78 (glucose-regulated protein 78 kDa). This unusual mechanism of regulating secretion from the constitutive secretory pathway provides a potentially missing link in the mechanism by which extracellular MANF protects cells from stresses that deplete SR/ER calcium. Consistent with this was our finding that administration of recombinant MANF to mice decreased tissue damage in an in vivo model of myocardial infarction, a condition during which ER calcium is known to be dysregulated, and MANF expression is induced.


European propolis is highly active against trypanosomatids including Crithidia fasciculata.
Extracts of 35 samples of European propolis were tested against wild type and resistant strains of the protozoal pathogens Trypanosoma brucei, Trypanosoma congolense and Leishmania mexicana. The extracts were also tested against Crithidia fasciculata a close relative of Crithidia mellificae, a parasite of bees. Crithidia, Trypanosoma and Leishmania are all members of the order Kinetoplastida. High levels of activity were obtained for all the samples with the levels of activity varying across the sample set. The highest levels of activity were found against L. mexicana. The propolis samples were profiled by using liquid chromatography with high resolution mass spectrometry (LC-MS) and principal components analysis (PCA) of the data obtained indicated there was a wide variation in the composition of the propolis samples. Orthogonal partial least squares (OPLS) associated a butyrate ester of pinobanksin with high activity against T. brucei whereas in the case of T. congolense high activity was associated with methyl ethers of chrysin and pinobanksin. In the case of C. fasciculata highest activity was associated with methyl ethers of galangin and pinobanksin. OPLS modelling of the activities against L. mexicana using the mass spectrometry produced a less successful model suggesting a wider range of active components.


Anatomical findings of the vanishing testis.
Inguinal exploration of 62 clinically impalpable undescended testes reveals 40 (64%) instances of absent testis. Nine (22.5%) of the 40 explorations showed complete absence of the testes along with the epididymis and vas deferens, six (15%) were associated with only vas ending blindly, and the other 25 (67.5%) were associated with blind-ending vas deferens and blood vessels: the "vanishing testis syndrome." The anatomical findings of these vanishing testes showed the presence of a vas deferens and vessels lying side by side in the inguinal canal. In Addition, two patients with bilateral vanishing testis were found to be a normal 46 xy phenotypic male. Since testicular tissue is necessary in utero to stimulate external genital development, these findings, imply that testes vanished at some point following induction of masculinization, possibly as a result of an intrauterine vascular accident or testicular torsion. Also, according to the anatomical findings of vanishing testis, it gave us the signal to terminate the exploration when we found absent testis with a blind-ending vas plus vessels on one or both side in an operation for impalpable testis.


LINC00337 Regulates KLF5 and Maintains Stem-Cell Like Traits of Cervical Cancer Cells by Modulating miR-145.
Accumulating literature and evidence has highlighted the cancer stem-like cell (CSC) model as a cellular mechanism responsible for the phenotypic heterogeneity observed in various types of cancers, including cervical cancer. Long non-coding RNAs (lncRNAs) have been implicated in the retention of stem cell-like traits in cancer cells. However, the role of lncRNAs in the acquisition and maintenance of CSCs in cervical cancer remains largely unknown. Hence, the current study identified that LINC00337 knockdown diminished the CSC-like properties of CD44+/CD24low/-SFCs, evidenced by a decline in the generation of tumorospheres and colonies, a reduction in multi-drug resistance gene-1 (MDR-1), Nanog, Sox2, and Oct4 expression, along with an enhancement in cell apoptosis. RNA pull-down assays and RNA immunoprecipitation revealed the role of LINC00337 as a competing endogenous RNA (ceRNA) of microRNA-145 (miR-145). Furthermore, the miR-145 mRNA target, Kruppel-like factor 5 (KLF5), was decreased in CD44+/CD24low/-SFCs upon LINC00337 knockdown. The in vitro results were reproduced in in vivo studies, which provided verification attesting that LINC00337 knockdown attenuated the tumorigenicity of CD44+/CD24low/-SFCs in nude mice. Taken together, the key findings of the current study demonstrate that LINC00337 acts as an oncogenic lncRNA in cervical cancer and exerts its influence on the expression of KLF5 and the maintenance of cancer stem cell-like properties by means of downregulating miR-145.


Predicting Axillary Lymph Node Metastasis in Early Breast Cancer Using Deep Learning on Primary Tumor Biopsy Slides.
To develop and validate a deep learning (DL)-based primary tumor biopsy signature for predicting axillary lymph node (ALN) metastasis preoperatively in early breast cancer (EBC) patients with clinically negative ALN. A total of 1,058 EBC patients with pathologically confirmed ALN status were enrolled from May 2010 to August 2020. A DL core-needle biopsy (DL-CNB) model was built on the attention-based multiple instance-learning (AMIL) framework to predict ALN status utilizing the DL features, which were extracted from the cancer areas of digitized whole-slide images (WSIs) of breast CNB specimens annotated by two pathologists. Accuracy, sensitivity, specificity, receiver operating characteristic (ROC) curves, and areas under the ROC curve (AUCs) were analyzed to evaluate our model. The best-performing DL-CNB model with VGG16_BN as the feature extractor achieved an AUC of 0.816 (95% confidence interval (CI): 0.758, 0.865) in predicting positive ALN metastasis in the independent test cohort. Furthermore, our model incorporating the clinical data, which was called DL-CNB+C, yielded the best accuracy of 0.831 (95%CI: 0.775, 0.878), especially for patients younger than 50 years (AUC: 0.918, 95%CI: 0.825, 0.971). The interpretation of DL-CNB model showed that the top signatures most predictive of ALN metastasis were characterized by the nucleus features including density (p = 0.015), circumference (p = 0.009), circularity (p = 0.010), and orientation (p = 0.012). Our study provides a novel DL-based biomarker on primary tumor CNB slides to predict the metastatic status of ALN preoperatively for patients with EBC.


Additivity of molecular fields: CoMFA study on dual activators of PPARalpha and PPARgamma.
Recent trends in drug discovery include methods to identify dual and triple activating drugs. This approach is being successfully employed in malaria, cancer, asthma, insulin resistance, etc. Molecular field analysis has been employed in correlating pharmacological data and field parameters. In this paper we introduce the concept of additivity of molecular fields to correlate molecular fields of dual activators and their pIC(50) values. PPARalpha and PPARgamma dual activators, which affect hypertriglyceridemia and hyperglycemia, have been chosen to validate the molecular field additivity concept. Three CoMFA models namely alpha-model, gamma-model and dual-model have been developed. The validity of this concept has been ascertained by (a) comparing contour maps, (b) by comparing CoMFA results with FlexX docking results and (c) by analyzing newly designed molecules.


Pre-eclampsia: pathophysiology and clinical implications.
Pre-eclampsia is a common disorder that particularly affects first pregnancies. The clinical presentation is highly variable but hypertension and proteinuria are usually seen. These systemic signs arise from soluble factors released from the placenta as a result of a response to stress of syncytiotrophoblast. There are two sub-types: early and late onset pre-eclampsia, with others almost certainly yet to be identified. Early onset pre-eclampsia arises owing to defective placentation, whilst late onset pre-eclampsia may center around interactions between normal senescence of the placenta and a maternal genetic predisposition to cardiovascular and metabolic disease. The causes, placental and maternal, vary among individuals. Recent research has focused on placental-uterine interactions in early pregnancy. The aim now is to translate these findings into new ways to predict, prevent, and treat pre-eclampsia.


The proteolytic processing site of the precursor of lysyl oxidase.
The precise cleavage site of the N-terminal propeptide region of the precursor of lysyl oxidase has not yet been established, due to N-terminal blocking of the mature protein. Using a combination of peptide fragmentation, amino acid sequencing, time-of-flight m.s. and partial chemical unblocking procedures, it is shown that the mature form of lysyl oxidase begins at residue Asp-169 of the precursor protein (numbered according to the human sequence). The cleavage site is 28 residues to the C-terminal side of the site previously suggested on the basis of apparant molecular mass by SDS/PAGE, with the consequence that the two putative, N-linked glycosylation sites and the position of the Arg/Gln sequence polymorphism are now all in the precursor region.


A new intertype recombinant between genotypes C and D of hepatitis B virus identified in China.
Hepatitis B virus (HBV) genotypes have a characteristic geographical distribution. More than 90% of chronic HBV patients in China are infected with genotypes B or C. Here, eight HBV isolates that were initially classified as genotype D by PCR-restriction fragment length polymorphism analysis were analysed in detail. The complete HBV genome was sequenced and compared with 32 sequences retrieved from GenBank, representing HBV genotypes A-G. Phylogenetic analysis of the S gene (nt 10-800) classified all eight isolates as genotype D. However, phylogenetic analyses of nt 800-10 and the open reading frames (ORFs) of the precore/core and X genes classified all eight isolates as genotype C. This discordance between phylogenetic trees reconstructed on different ORFs suggested that intertype recombination has occurred in all eight isolates. By using the simplot program, the site of recombination with genotype D was located in the preS2/S region, spanning nt 10-799 in seven of eight isolates and nt 10-1499 in the other isolate. These results demonstrate that intertype recombination should be considered as a type of variation that increases the genetic diversity of HBV. Hybrids of different HBV genotypes might exhibit specific virological properties and their significance in the diagnosis and management of chronic hepatitis B deserves further investigation.


Biological Response Modifiers Programme and cancer chemotherapy.
The Biological Response Modifiers Programme (BRMP) is a comprehensive programme of the National Cancer Institute to evaluate the therapeutic effect of biological substances that might be useful in the treatment of cancer. Immunomodulation agents, interferons, cytokines, growth and maturation factors, antitumour effector cells and antibodies, and chalones are all potential biological response modifiers (BRM). The BRMP has a biologic preclinical evaluation capability whereby new agents can be evaluated as potential BRM in preclinical models. Effective biological response modifying activity in preclinical models with a therapeutic efficacy in cancer is a requirement for an agent to continue on into clinical testing. Initial results with interferon, thymic factors and other immune modulators are interesting but very preliminary. These agents are being tested for the ability to alter biological responses as well as for toxic and therapeutic effects. Optimization of the biological response modification capability may be predictive for therapeutic efficacy. Purified cytokines, specific tumour-associated antigens, and monoclonal antibody directed toward antigenic specificiteis on the cancer cell all provide exciting avenues for future investigation. Components of the BRMP will be described and preliminary results with selected BRM will be presented. The interaction between BRM and cancer chemotherapy should be investigated further. Certain cytokines are known to enhance bone-marrow-stem cell function and many of these may be useful in conjunction with cancer chemotherapy to lessen the toxic on normal cell populations often encountered with chemotherapy.


Accumulation of UDP-glucose pyrophosphorylase by axenically grown amoebae of Dictyostelium discoideum.
Amoebae of the slime mould Dictyostelium discoideum AX2 possess only low UDP-glucose pyrophosphorylase activity when grown on autoclaved Klebsiella aerogenes (approx. 30 units/mg of protein), but accumulate the enzyme to approx. 150-200 units/mg of protein during vegetative growth in axenic medium. The vegetative accumulation of UDP-glucose pyrophosphorylase by axenically grown cells is prevented if autoclaved K. aerogenes are included in the axenic medium, suggesting the absence of a specific inducer. Affinity chromatography using anti-(UDP-glucose pyrophosphorylase) antibody and sodium dodecyl sulphate/polyacrylamide-gel electrophoresis indicate that the enzyme accumulated during axenic growth and that normally accumulated during development are immunologically cross-reactive and that both are composed of two subunits with mol.wts. 55,600 and 57,500 present in approximately equal amounts in the active enzyme.


Tissue-type-specific transcriptome analysis identifies developing xylem-specific promoters in poplar.
Plant biotechnology offers a means to create novel phenotypes. However, commercial application of biotechnology in crop improvement programmes is severely hindered by the lack of utility promoters (or freedom to operate the existing ones) that can drive gene expression in a tissue-specific or temporally controlled manner. Woody biomass is gaining popularity as a source of fermentable sugars for liquid fuel production. To improve the quantity and quality of woody biomass, developing xylem (DX)-specific modification of the feedstock is highly desirable. To develop utility promoters that can drive transgene expression in a DX-specific manner, we used the Affymetrix Poplar Genome Arrays to obtain tissue-type-specific transcriptomes from poplar stems. Subsequent bioinformatics analysis identified 37 transcripts that are specifically or strongly expressed in DX cells of poplar. After further confirmation of their DX-specific expression using semi-quantitative PCR, we selected four genes (DX5, DX8, DX11 and DX15) for in vivo confirmation of their tissue-specific expression in transgenic poplars. The promoter regions of the selected DX genes were isolated and fused to a β-glucuronidase (GUS)-reported gene in a binary vector. This construct was used to produce transgenic poplars via Agrobacterium-mediated transformation. The GUS expression patterns of the resulting transgenic plants showed that these promoters were active in the xylem cells at early seedling growth and had strongest expression in the developing xylem cells at later growth stages of poplar. We conclude that these DX promoters can be used as a utility promoter for DX-specific biomass engineering.


Detecting S-adenosyl-L-methionine-induced conformational change of a histone methyltransferase using a homogeneous time-resolved fluorescence-based binding assay.
A homogeneous time-resolved fluorescence (HTRF)-based binding assay has been established to measure the binding of the histone methyltransferase (HMT) G9a to its inhibitor CJP702 (a biotin analog of the known peptide-pocket inhibitor, BIX-01294). This assay was used to characterize G9a inhibitors. As expected, the peptide-pocket inhibitors decreased the G9a-CJP702 binding signal in a concentration-dependent manner. In contrast, the S-adenosyl-L-methionine (SAM)-pocket compounds, SAM and sinefungin, significantly increased the G9a-CJP702 binding signal, whereas S-adenosyl-L-homocysteine (SAH) showed minimal effect. Enzyme kinetic studies showed that CJP702 is an uncompetitive inhibitor (vs. SAM) that has a strong preference for the E:SAM form of the enzyme. Other data presented suggest that the SAM/sinefungin-induced increase in the HTRF signal is secondary to an increased E:SAM or E:sinefungin concentration. Thus, the G9a-CJP702 binding assay not only can be used to characterize the peptide-pocket inhibitors but also can detect the subtle conformational differences induced by the binding of different SAM-pocket compounds. To our knowledge, this is the first demonstration of using an uncompetitive inhibitor as a probe to monitor the conformational change induced by compound binding with an HTRF assay.


Performance-enhancing sports supplements: role in critical care.
Many performance-enhancing supplements and/or drugs are increasing in popularity among professional and amateur athletes alike. Although the uncontrolled use of these agents can pose health risks in the general population, their clearly demonstrated benefits could prove helpful to the critically ill population in whom preservation and restoration of lean body mass and neuromuscular function are crucial. Post-intensive care unit weakness not only impairs post-intensive care unit quality of life but also correlates with intensive care unit mortality. This review covers a number of the agents known to enhance athletic performance, and their possible role in preservation of muscle function and prevention/treatment of post-intensive care unit weakness in critically ill patients. These agents include testosterone analogues, growth hormone, branched chain amino acid, glutamine, arginine, creatine, and beta-hydryoxy-beta-methylbutyrate. Three of the safest and most effective agents in enhancing athletic performance in this group are creatine, branched-chain amino acid, and beta-hydryoxy-beta-methylbutyrate. However, these agents have received very little study in the recovering critically ill patient suffering from post-intensive care unit weakness. More placebo-controlled studies are needed in this area to determine efficacy and optimal dosing. It is very possible that, under the supervision of a physician, many of these agents may prove beneficial in the prevention and treatment of post-intensive care unit weakness.


Variations in culture pH affect the cloning efficiency and differentiation of progenitor cells in ex vivo haemopoiesis.
Haemopoietic cultures may experience pH variations of as much as 0.5 units depending on culture duration and cell density. Since pH is a potent modulator of cellular proliferation and differentiation, we examined its effects on the performance of both semisolid and liquid haemopoietic cultures. Culture pH was found to have substantial effects both on progenitor cloning efficiency (as measured in liquid cultures) and on progenitor cell differentiation (as measured in methylcellulose cultures). Liquid cultures were conducted with both peripheral blood (PB) mononuclear cells (MNCs) and cord blood (CB) MNCs using growth factor combinations that promote either erythroid expansion (IL-3/IL-6/SCF/Epo) or granulocyte/macrophage expansion (IL-3/IL-6/SCF/G-CSF/GM-CSF). Reduced pH was found to have either a positive or neutral effect on the expansion and cloning efficiency of progenitors in ex vivo liquid cultures. Cloning efficiencies of PB BFU-E in the erythroid combination were 9-fold higher at low pH (7.1) when compared to high pH (7.6). A small pH increase of 0.2 units over physiological values consistently produced significant reductions (42-85%) in cloning efficiencies for all cell types and cytokine combinations tested. Methylcellulose cultures conducted using CB MNC and PB MNC indicated that differentiation of CFU-GM into progeny was optimal between pH 7.2 and 7.4. The differentiation of erythroid progenitors (BFU-E) progressively increased as pH was increased from 6.95 (no colonies detected) to 7.4 (maximum colonies detected), to 7.6 (maximum haemoglobin content). Methylcellulose cultures using PB CD34+ cells exhibited similar patterns to the MNC cultures. We conclude that even small variations in pH substantially affected the performance of human haemopoietic cultures. The erythroid lineage was particularly sensitive, with its extent of differentiation increasing with increasing pH. PB progenitors are more sensitive to pH variations than CB progenitors.


[Segmental and atypical resections in lung cancer].
Segmental or atypical resection was performed in 25 out of 1120 (2.2%) patients operated on for lung cancer. The choice was determined by concomitant pathology and old age. No fatalities were registered in the postoperative period. Five-year survival in the segmental and atypical resection group was 33.3%. The procedure discussed may be recommended if radical surgery is contraindicated.


Should I stay or should I go? Shedding of RPTPs in cancer cells switches signals from stabilizing cell-cell adhesion to driving cell migration.
Dissolution of cell-cell adhesive contacts and increased cell-extracellular matrix adhesion are hallmarks of the migratory and invasive phenotype of cancer cells. These changes are facilitated by growth factor binding to receptor protein tyrosine kinases (RTKs). In normal cells, cell-cell adhesion molecules (CAMs), including some receptor protein tyrosine phosphatases (RPTPs), antagonize RTK signaling by promoting adhesion over migration. In cancer, RTK signaling is constitutive due to mutated or amplified RTKs, which leads to growth factor independence, or autonomy. An alternative route for a tumor cell to achieve autonomy is to inactivate cell-cell CAMs such as RPTPs. RPTPs directly mediate cell adhesion and regulate both cadherin-dependent adhesion and signaling. In addition, RPTPs antagonize RTK signaling by dephosphorylating molecules activated following ligand binding. Both RPTPs and cadherins are downregulated in tumor cells by cleavage at the cell surface. This results in shedding of the extracellular, adhesive segment and displacement of the intracellular segment, altering its subcellular localization and access to substrates or binding partners. In this commentary we discuss the signals that are altered following RPTP and cadherin cleavage to promote cell migration. Tumor cells both step on the gas (RTKs) and disconnect the brakes (RPTPs and cadherins) during their invasive and metastatic journey.


Cosmic Birefringence from the Planck Data Release 4.
We search for the signature of parity-violating physics in the cosmic microwave background, called cosmic birefringence, using the Planck data release 4. We initially find a birefringence angle of β=0.30°±0.11° (68% C.L.) for nearly full-sky data. The values of β decrease as we enlarge the Galactic mask, which can be interpreted as the effect of polarized foreground emission. Two independent ways to model this effect are used to mitigate the systematic impact on β for different sky fractions. We choose not to assign cosmological significance to the measured value of β until we improve our knowledge of the foreground polarization.


[Adjustment of the theory of self-care deficit in home care in the light of Barnum's model].
The study aimed at analyzing the Self-Care Deficit Theory adjustment, focusing on the atmosphere and the home care provider. Authors adopted the Barnum's model that suggests questions on the use of its concepts, seeking for its interrelations and scope. The answers to those questions were based on the Self-Care Deficit Theory applied to a real situation, observed in home care visits occurred in the second semester of 2000 to a paraplegic patient, whose care was provided by this wife and the family health nursing team. Authors found minimal adjustment conditions for home care, considering the environment and the care providers. They considered, comparatively, that the chosen theory may be directed to the care of patients in home hospitalization under the supervision of a nurse.


Biosynthesis of HSAF, a tetramic acid-containing macrolactam from Lysobacter enzymogenes.
HSAF was isolated from Lysobacter enzymogenes , a bacterium used in the biological control of fungal diseases of plants. Structurally, it is a tetramic acid-containing macrolactam fused to a tricyclic system. HSAF exhibits a novel mode of action by disrupting sphingolipids important to the polarized growth of filamentous fungi. Here we describe the HSAF biosynthetic gene cluster, which contains only a single-module polyketide synthase/nonribosomal peptide synthetase (PKS/NRPS), although the biosynthesis of HSAF apparently requires two separate polyketide chains that are linked together by one amino acid (ornithine) via two amide bonds. Flanking the PKS/NRPS are six genes that encoding a cascade of four tightly clustered redox enzymes on one side and a sterol desaturase/fatty acid hydroxylase and a ferredoxin reductase on the other side. The genetic data demonstrate that the four redox genes, in addition to the PKS/NRPS gene and the sterol desaturase/fatty acid hydroxylase gene, are required for HSAF production. The biochemical data show that the adenylation domain of the NRPS specifically activates L-ornithine and that the four-domain NRPS is able to catalyze the formation of a tetramic acid-containing product from acyl-S-ACP and ornithinyl-S-NRPS. These results reveal a previously unrecognized biosynthetic mechanism for hybrid PK/NRP in prokaryotic organisms.


Identification of exercise-regulated genes in mice exposed to cigarette smoke.
Cigarette smoke (CS) is the major risk factor for COPD and is linked to cardiopulmonary dysfunction. Exercise training as part of pulmonary rehabilitation is recommended for all COPD patients. It has several physiological benefits, but the mechanisms involved remain poorly defined. Here, we employed transcriptomic profiling and examined lung endothelium to investigate novel interactions between exercise and CS on cardiopulmonary alterations. Mice were exposed to 20 weeks of CS, CS + 6 weeks of high-intensity interval training on a treadmill, or control. Lung and cardiac (left and right ventricle) tissue were harvested and RNA-sequencing was performed and validated with RT-qPCR. Immunohistochemistry assessed pulmonary arteriolar changes. Transcriptome analysis between groups revealed 37 significantly regulated genes in the lung, 21 genes in the left ventricle, and 43 genes in the right ventricle (likelihood-ratio test). Validated genes that showed interaction between exercise and CS included angiotensinogen (p = 0.002) and resistin-like alpha (p = 0.019) in left ventricle, with prostacyclin synthetase different in pulmonary arterioles (p = 0.004). Transcriptomic profiling revealed changes in pulmonary and cardiac tissue following exposure to CS, with exercise training exerting rescue effects. Exercise-regulated genes included angiotensinogen and resistin-like alpha, however, it remains unclear if these represent potential candidate genes or biomarkers that could play a role during pulmonary rehabilitation.


Role of Terlipressin and Albumin for Hepatorenal Syndrome in Liver Transplantation.
Hepatorenal syndrome (HRS) is one of the most ominous complications of portal hypertension in patients with decompensated cirrhosis and ascites. It is associated with very high mortality on the wait list. Liver transplantation (LT) is the most successful therapeutic option for patients with HRS. However, not all the LT candidates with HRS are able to receive a deceased donor allograft in a timely manner because it is a scarce resource and patients may need alternative best supportive treatment with systemic splanchnic vasoconstrictors and albumin as a bridge to transplant. The combination of terlipressin and albumin is efficacious in the reversal of HRS and is used worldwide. More recently, the multicenter, randomized, placebo-controlled double-blind study to confirm efficacy and safety of terlipressin in subjects with hepatorenal syndrome type 1 (the CONFIRM study) trial demonstrated the efficacy of terlipressin and albumin in the reversal of HRS in a North American cohort. The aim of this article is to review the role of terlipressin and albumin in LT candidates with HRS in the United States.


Beam instabilities in the scale-free regime.
The instabilities arising in a one-dimensional beam sustained by the diffusive photorefractive nonlinearity in out-of-equilibrium ferroelectrics are theoretically and numerically investigated. In the "scale-free model," in striking contrast with the well-known spatial modulational instability, two different beam instabilities dominate: a defocusing and a fragmenting process. Both are independent of the beam power and are not associated with any specific periodic pattern.


Investigating Virus-Host Interactions in Cultured Primary Honey Bee Cells.
Honey bee (Apis mellifera) health is impacted by viral infections at the colony, individual bee, and cellular levels. To investigate honey bee antiviral defense mechanisms at the cellular level we further developed the use of cultured primary cells, derived from either larvae or pupae, and demonstrated that these cells could be infected with a panel of viruses, including common honey bee infecting viruses (i.e., sacbrood virus (SBV) and deformed wing virus (DWV)) and an insect model virus, Flock House virus (FHV). Virus abundances were quantified over the course of infection. The production of infectious virions in cultured honey bee pupal cells was demonstrated by determining that naïve cells became infected after the transfer of deformed wing virus or Flock House virus from infected cell cultures. Initial characterization of the honey bee antiviral immune responses at the cellular level indicated that there were virus-specific responses, which included increased expression of bee antiviral protein-1 (GenBank: MF116383) in SBV-infected pupal cells and increased expression of argonaute-2 and dicer-like in FHV-infected hemocytes and pupal cells. Additional studies are required to further elucidate virus-specific honey bee antiviral defense mechanisms. The continued use of cultured primary honey bee cells for studies that involve multiple viruses will address this knowledge gap.


Kalirin expression is regulated by multiple promoters.
Kalirin is a multidomain guanine nucleotide exchange factor for small GTPbinding proteins of the Rho family. It is expressed in multiple isoforms that contain different combinations of functional domains and display a complex pattern of expression during brain development. In addition to the isoforms generated through alternative splicing, we have identified multiple transcriptional start sites in rats and humans. These multiple transcriptional start sites result in full-length Kalirin transcripts possessing different 5' ends encoding proteins with differing amino termini. These alternative first exons display different patterns of expression in developing rats and humans and in cultured cells. Most of these alternate first exons lie >100 kb upstream of exon 2 in both rats and humans. Comparisons of the rat and human Kalirin promoter regions reveal numerous shared potential regulatory elements.


Assembly stoichiometry of the GluK2/GluK5 kainate receptor complex.
Ionotropic glutamate receptors assemble as homo- or heterotetramers. One well-studied heteromeric complex is formed by the kainate receptor subunits GluK2 and GluK5. Retention motifs prevent trafficking of GluK5 homomers to the plasma membrane, but coassembly with GluK2 yields functional heteromeric receptors. Additional control over GluK2/GluK5 assembly seems to be exerted by the aminoterminal domains, which preferentially assemble into heterodimers as isolated domains. However,the stoichiometry of the full-length GluK2/GluK5 receptor complex has yet to be determined, as is the case for all non-NMDA glutamate receptors. Here, we address this question, using a single-molecule imaging technique that enables direct counting of the number of each GluK subunit type in homomeric and heteromeric receptors in the plasma membranes of live cells. We show that GluK2 and GluK5 assemble with 2:2 stoichiometry. This is an important step toward understanding the assembly mechanism, architecture, and functional consequences of heteromer formation in ionotropic glutamate receptors.


[Magnetic resonance imaging of the breasts: perspectives in clinical use].
Magnetic resonance imaging (MRI) of the breasts was introduced in clinical practice over 10 years ago. The method is based on visualization of morphology of the lesion and pathophysiology of their vascularization. It is useful complementary method to mammography and breasts ultrasound, with potential to detect clinically or mammography occult lesions. Compared to the conventional visualization methods, MRI of the breasts is more sensitive, although specificity of the method is lower than sensitivity. The indications for MRI of the breasts are: differentiation between recurrent malignant disease and postoperative or post irradiation changes, local cancer staging in preoperative planning, breasts implants, axillary metastases with occult primary tumor and cases with inconclusive conventional imaging. The limitations of MRI of the breasts are: low sensitivity for in situ carcinomas, false negative results for 10% carcinomas and hormone-induced false positive results. The further development of the method includes: breasts cancer screening in the patients with high risk, monitoring of the effects of neo-adjuvant chemotherapy, MR-guided biopsy and new contrast agents.


Transient loss and recovery of oral chemesthesis, taste and smell with COVID-19: a small case-control series.
Anosmia is common with respiratory virus infections, but loss of taste or chemesthesis is rare. Reports of true taste loss with COVID-19 were viewed skeptically until confirmed by multiple studies. Nasal menthol thresholds are elevated in some with prior COVID-19 infections, but data on oral chemesthesis are lacking. Many patients recover quickly, but precise timing and synchrony of recovery are unclear. Here, we collected broad sensory measures over 28 days, recruiting adults (18-45 years) who were COVID-19 positive or recently exposed (close contacts per U.S. CDC criteria at the time of the study) in the first half of 2021. Participants received nose clips, red commercial jellybeans (Sour Cherry and Cinnamon), and scratch-n-sniff cards (ScentCheckPro). Among COVID-19 cases who entered the study on or before Day 10 of infection, Gaussian Process Regression showed odor identification and odor intensity (two distinct measures of function) each declined relative to controls (close contacts who never developed COVID-19), but effects were larger for intensity than identification. To assess changes during early onset, we identified four COVID-19 cases who enrolled on or prior to Day 1 of their illness â€" this allowed for visualization of baseline ratings, loss, and recovery of function over time. Four controls were matched for age, gender, and race. Variables included sourness and sweetness (Sour Cherry jellybeans), oral burn (Cinnamon jellybeans), mean orthonasal intensity of four odors (ScentCheckPro), and perceived nasal blockage. Data were plotted over 28 days, creating panel plots for the eight cases and controls. Controls exhibited stable ratings over time. By contrast, COVID-19 cases showed sharp deviations over time. No single pattern of taste loss or recovery was apparent, implying different taste qualities might recover at different rates. Oral burn was transiently reduced for some before recovering quickly, suggesting acute loss may be missed in data collected after acute illness ends. Changes in odor intensity or odor identification were not explained by nasal blockage. Collectively, intensive daily testing shows orthonasal smell, oral chemesthesis and taste were each altered by acute COVID-19 infection, and this disruption was dyssynchronous for different modalities, with variable loss and recovery rates across modalities and individuals.


Role of Private Sector in Providing Tuberculosis Care: Evidence from a Population-based Survey in India.
In India, a large segment of the population seeks health care services from individual or institutional private health-care providers for health care. We analyzed a nationally representative data to identify the role of private providers in delivering health care for patients with tuberculosis. The primary data source for the present analysis was the 60(th) round of the National Sample Survey. Distribution frequencies were used to analyze the distribution of key sociodemographic variables and multiple logistic regression was used to analyze the association between these variables and healthcare seeking behavior. A sample of 2203 respondents who had received ambulatory care for tuberculosis, and 4568 respondents who had received inpatient treatment were analyzed. About half of the respondents had attended private facilities for TB care. Sociodemographic variables such as paediatric age group, females, higher level of education, and economic groups were associated with attendance at private sector. Dissatisfaction with services in government facilities was cited as the main reason for preferring private facilities. Private providers play an important role in providing health care services to a large proportion of patients with tuberculosis. There is a need for innovative measures to increase participation of the private sector in the national TB control program and to improve the quality of services in government facilities.


Lack of secondary hyperalgesia and central sensitization in an acute sheep model.
We aimed to determine the following in an experimental acute pain model in sheep: (1) whether multimodal analgesia with intravenous fentanyl and ketorolac was more effective than fentanyl alone; (2) whether secondary hyperalgesia (central sensitization) occurred in adjacent (foreleg) dermatomes after thoracic surgery; (3) whether ketorolac used preemptively influenced the development of secondary hyperalgesia after surgery. Changes in primary nociception were measured by increases to tolerated pressure, applied to the foreleg by a blunt pin, before foreleg withdrawal occurred. Changes to breath-to-breath interval and estimated end-tidal CO2 were used as indices of respiratory effects. Study 1 (n = 6) compared the paired responses to acute nociception after ketorolac (90 mg) or saline (control) pretreatment, followed by fentanyl (graded, 0 mg to 1.5 mg). Study 2 (n = 6) used a cross-over of ketorolac (90 mg) or saline (control) 24 hours and 1 hour, respectively, before a standardized thoracotomy incision, followed by antinociceptive testing with ketorolac (90 mg) and fentanyl (0.6 mg) daily over 4 days. In study 1, fentanyl produced naloxone-antagonizable antinociception and respiratory depression. Ketorolac did not affect fentanyl antinociception, except for prolonging antinociception at the highest dose; it did not affect the respiratory effects. In study 2, preemptive ketorolac had no effect on the postoperative antinociceptive or respiratory effects of fentanyl. The pharmacokinetics of fentanyl were unaltered by ketorolac. The results obtained in this acute pain model found no significant evidence of a fentanyl-ketorolac interaction, of central sensitization as shown by secondary hyperalgesia, or of a preemptive analgesic effect.


Accelerating translational research by clinically driven development of an informatics platform--a case study.
Translational medicine is becoming increasingly dependent upon data generated from health care, clinical research, and molecular investigations. This increasing rate of production and diversity in data has brought about several challenges, including the need to integrate fragmented databases, enable secondary use of patient clinical data from health care in clinical research, and to create information systems that clinicians and biomedical researchers can readily use. Our case study effectively integrates requirements from the clinical and biomedical researcher perspectives in a translational medicine setting. Our three principal achievements are (a) a design of a user-friendly web-based system for management and integration of clinical and molecular databases, while adhering to proper de-identification and security measures; (b) providing a real-world test of the system functionalities using clinical cohorts; and (c) system integration with a clinical decision support system to demonstrate system interoperability. We engaged two active clinical cohorts, 747 psoriasis patients and 2001 rheumatoid arthritis patients, to demonstrate efficient query possibilities across the data sources, enable cohort stratification, extract variation in antibody patterns, study biomarker predictors of treatment response in RA patients, and to explore metabolic profiles of psoriasis patients. Finally, we demonstrated system interoperability by enabling integration with an established clinical decision support system in health care. To assure the usefulness and usability of the system, we followed two approaches. First, we created a graphical user interface supporting all user interactions. Secondly we carried out a system performance evaluation study where we measured the average response time in seconds for active users, http errors, and kilobits per second received and sent. The maximum response time was found to be 0.12 seconds; no server or client errors of any kind were detected. In conclusion, the system can readily be used by clinicians and biomedical researchers in a translational medicine setting.


Degranulation, membrane addition, and shape change during chemotactic factor-induced aggregation of human neutrophils.
Neutrophils stimulated by the chemotactic factor formyl-methionyl-leucyl-phenyl-alanine (FMLP) undergo a transient change in surface properties that permits the cells to adhere more readily to surfaces and to each other. This transient change can be monitored by light scattering as stimulated neutrophils form aggregates while stirred in a platelet aggregometer. Maximum change in light scattering occurs within 1 min and correlates with an increase in the percentage of cells that are in aggregates of four or more cells and a decrease in the percentage of single cells. With time (3-5 min), small aggregates disappear and single cells reappear. The transient change in adhesiveness is accompanied by a persistent change in cell shape; the cells become polarized and protrude ruffles from one sector of the cell surface. During aggregation the cells adhere to one another with smooth sides together and ruffles pointed outward. During disaggregation the cells dissociate laterally with the simultaneous internalization of membrane in the region opposite the ruffles. Particle bound to the surface by charge (thorotrast, cationized ferritin) are concentrated and internalized in this region. The change in cell shape from round to ruffled occurs within seconds, suggesting that membrane is added to the cell surface from an intracellular store. We therefore quantified surface membrane by electron microscopy morphometry and measured a 25% increase within 10 s of adding FMLP. The source of new membrane appeared to be the specific granule membrane since the kinetics of granule discharge (between 30% and 50% of all release occurs in the first 10 s) correlate with the appearance of new membrane. Furthermore, the amount of membrane that appears at the cell surface at 10 s correlates with that lost from intracellular granules in that time. Chemotaxin-induced aggregation thus begins with granule discharge and membrane addition followed by protrusion of ruffles. Adherence is maximal at 60 s and the gradual loss of adhesiveness that follows is associated with uropod formation and enhanced endocytic activity.


Neurologic Complications: Effects of Ipilimumab and Nivolumab Therapy in Patients With Metastatic Melanoma.
Checkpoint inhibitors are a form of immunotherapy that have revolutionized treatment for malignant melanoma, resulting in longer survival and better disease control. Multiple autoimmune disorders can occur with the use of checkpoint inhibitors, including severe, potentially fatal neurologic complications. Although neurologic complications are uncommon, their early recognition and treatment is required. The purpose of this article is to present information on neurologic complications of ipilimumab and nivolumab to inform nursing practice. Recommendations for evaluation and treatment of neurologic complications are reviewed.


Surgical management of the spinal deformity in Ehlers-Danlos syndrome type VI.
Five patients with Ehlers-Danlos syndrome type VI were surgically treated for their spinal deformities. All of them were female. All but one had a double-major thoracic and lumbar curve. One had a mild scoliotic curve but severe thoracic lordosis. Anterior discectomy and fusion and posterior instrumentation was performed in a single stage in two patients, while two had anterior and posterior surgery performed in two stages. The remaining patient underwent posterior surgery only, because of the relative mildness of the deformity and adequate flexibility. Two serious vascular complications were confronted. One patient experienced avulsion of segmental arteries and rupture of iliac artery and vein during anterior surgery. Another patient had avulsion of superior gluteal artery, which happened during subperiosteal dissection to harvest iliac bone graft. Fortunately, we did not see any temporary or permanent neurological complications. The spinal fusions appeared solid radiographically, with no implant failure and loss of correction after an average follow-up of 4 years.


Stimulus-reinforcement-based decision making and anxiety: impairment in generalized anxiety disorder (GAD) but not in generalized social phobia (GSP).
Generalized social phobia (GSP) involves the fear/avoidance of social situations whereas generalized anxiety disorder (GAD) involves an intrusive worry about everyday life circumstances. It remains unclear whether these, highly co-morbid, conditions represent distinct disorders or alternative presentations of a single underlying pathology. In this study, we examined stimulus-reinforcement-based decision making in GSP and GAD. Twenty unmedicated patients with GSP, 16 unmedicated patients with GAD and 19 age-, IQ- and gender-matched healthy comparison (HC) individuals completed the Differential Reward/Punishment Learning Task (DRPLT). In this task, the subject chooses between two objects associated with different levels of reward or punishment. Thus, response choice indexes not only reward/punishment sensitivity but also sensitivity to reward/punishment level according to between-object reinforcement distance. We found that patients with GAD committed a significantly greater number of errors than both the patients with GSP and the HC individuals. By contrast, the patients with GSP and the HC individuals did not differ in performance on this task. These results link GAD with anomalous non-affective-based decision making. They also indicate that GSP and GAD are associated with distinct pathophysiologies.


Murine cortactin is phosphorylated in response to fibroblast growth factor-1 on tyrosine residues late in the G1 phase of the BALB/c 3T3 cell cycle.
We have previously reported that BALB/c 3T3 cells require a prolonged exposure to fibroblast growth factor (FGF)-1 for the stimulation of maximal DNA synthesis, and this event correlates with the tyrosine phosphorylation of novel proteins late in G1 including a protein termed p80/p85 (Zhan, X., Hu, X., Friesel, R., and Maciag, T. (1993) J. Biol. Chem. 268, 9611-9620). We have purified, sequenced, and cloned the cDNA encoding p80/p85 and report that it is the murine homolog of the chicken cortactin gene and a member of the human hematopoietic specific-1 gene family. Immunochemical analysis of m-cortactin-tyrosine phosphorylation in response to FGF-1 demonstrates a biphasic phosphorylation pattern both as a weak immediate-early and strong mid to late G1 response protein. Because the chicken cortactin gene was originally isolated as a substrate for v-Src, FGF-1 may influence the enzymatic activity of other cell-associated tyrosine kinases which utilize p80/p85 (cortactin) as a polypeptide substrate.


[On the genus Nematodirus Ransom 1907 (Nematoda: Trichostrongyloidea)].
N. roscidus Railliet, 1911, from Dama dama (Italy); N. abnormalis May, 1920, from Ovis aries, Capra hircus and Rupicapra rupicapra (France, Italy, Australia); N. lanceolatus Ault, 1944, from Ovis aries (Iran); N. europaeus Jansen, 1972, from Capreolus caperolus and Ovis musimon (first record) (France); N. davtiani alpinus Biocca, Balbo & Costantini, 1974, from Rupicapra rupicapra (Italy); N. rupicaprae Biocca, Balbo & Costantini, 1983, from Rupicapra rupicapra (France); N. ibicis Biocca, Balbo & Costantini, 1983, from Capra ibex (Italy). In addition four new species are described: N. Bioccai n. sp. from Ovis aries (Iran), resembles N. lamae Becklund, 1963 from Lama pacos and Vicugna vicugna (Peru), N. ibicis Biocca, Balbo & Costantini, 1983 from Capra ibex (Italy), and N. archari Sokolova, 1948 from Ovis ammon (USSR), in that ray 8 is long and closely adpressed to ray 6 throughout most of its length. The new species differs from the first of the above in the form of the point of the spicules and from the remaining two since the two branches of the dorsal ray are well separated and the dorsal lobe well differentiated from the rest of the bursa. N. chabaudi n. sp. from Rupicapra rupicapra (France) is similar to N. oiratianus Rajewskaja, 1929, from Cervus canadensis asiaticus and Ovis aries (USSR), and N. europaeus Jansen, 1972 from Capreolus capreolus (Holland), in the disposition of its bursal rays. Il is distinguished mainly by the presence of three tubercles on the tail of the female. N. hugonnetae n. sp., from Rupicapra rupicapra (France), resembles N. bioccai n. sp. in the general form of the bursa and in that the spicules are fused at their extremities, but is distinguished by the form of the point of the spicules and by the disposition of the bursal bosses. N. iranicus n. sp. from Ovis aries (Iran), resembles N. hugonnetae n. sp. in the disposition of the bursal rays but is distinguished by having more cuticular ridges and a shorter ray 8. On the basis of morphology, host distribution and host paleogeography, an hypothesis is given on the evolution of the Nematodirus and the Nematodirinae. According to this hypothesis Nematodirus arose in North American Camelidae from an ancestor similar to lamanema (which occurs in neotropical Camelidae), later passed to nearctic Odocoileinae and Caprinae, and was transported secondarily into the palaearctic region with its hosts. Beginning with this genus, the Nematodirinae evolved in holarctic ruminants (Nematodirella), holarctic Leporidae (Nematodiroides and Rauschia) and palaearctic and oriental Ochotonidae (Murielus).


Cost analysis of methylprednisolone treatment of multiple sclerosis patients.
Intravenous methylprednisolone (IVMP) is the treatment of choice for multiple sclerosis (MS) patients undergoing acute exacerbation of disease symptoms and yet its cost has not been accurately determined. Determination of this cost in different settings is also pertinent to consideration of cost-saving alternatives to in-patient treatment. Cost analysis from the point of view of the health care system of IVMP treatment of MS patients receiving treatment in association with a selected Toronto teaching hospital in fiscal year 1994/95 was carried out. Costs of any concurrent treatments were excluded. Total cost for 92 patients, based on a 4 dose regime, was estimated to be $78,527. The the cost per patient was $1,1181.84 for in-patients (IP), $714.64 for out-patients of the MS Clinic (OP) and $774.21 for patients whose treatment was initiated in the Clinic, but completed in the home (HC). Sensitivity analyses indicated: 1) IP treatment was in all cases more expensive than that of OP or HC; 2) the cost savings of OP vs. HC was sensitive to assumptions made regarding Clinic overhead, Clinic nursing costs and Home Care Program overhead. Alternatives to in-patient care must be considered carefully. In this study, both out-patient and in-home treatment were cost-saving alternatives to in-patient treatment, but large differences in the cost of hospital out-patient vs. in-home care could not be demonstrated.


Thiourea reverses cross-links and restores biological activity in DNA treated with dichlorodiaminoplatinum (II).
Cis and trans dichlorodiaminoplatinum (II) compounds bind to DNA and form DNA cross-links, which are usually considered to be irreversible. Thiourea can reverse these cross-links without any apparent breakdown of the DNA. In addition, cis- and trans-Pt (II) treatment of lambda decreases its transfectivity. After suitable incubation with thiourea, full transfectivity of Pt(II)-treated lambda DNA can be restored.


The Role of Nitro (NO2-), Chloro (Cl), and Fluoro (F) Substitution in the Design of Antileishmanial and Antichagasic Compounds.
Neglected tropical diseases (NTDs) are responsible for over 500,000 deaths annually and are characterized by multiple disabilities. Leishmaniasis and Chagas diseases are among the most severe NTDs, and are caused by the Leishmania sp and Trypanosoma cruzi, respectively. Glucantime, pentamidine, and miltefosine are commonly used to treat leishmaniasis, whereas nifurtimox, benznidazole are current treatments for Chagas disease. However, these treatments are associated with drug resistance and severe side effects. Hence, the development of synthetic products, especially those containing N02, F, or Cl, are known to improve biological activity. The present work summarizes the information on the antileishmanial and antitrypanosomal activity of nitro-, chloro-, and fluorosynthetic derivatives. Scientific publications referring to halogenated derivatives in relation to antileishmanial and antitrypanosomal activities were hand-searched in databases such as SciFinder, Wiley, Science Direct, PubMed, ACS, Springer, Scielo, and so on. According to the literature information, more than 90 compounds were predicted as lead molecules with reference to their IC50/EC50 values in in vitro studies. It is worth mentioning that only active compounds with known cytotoxic effects against mammalian cells were considered in the present study. The observed activity was attributed to the presence of nitro-, fluoro-, and chloro-groups in the compound backbone. All in all, nitro and halogenated derivatives are active antileishmanial and antitrypanosomal compounds and can serve as the baseline for the development of new drugs against leishmaniasis and Chagas disease. However, efforts in in vitro and in vivo toxicity studies of the active synthetic compounds is still needed. Pharmacokinetic studies and the mechanism of action of the promising compounds need to be explored. The use of new catalysts and chemical transformation can afford unexplored halogenated compounds with improved antileishmanial and antitrypanosomal activity.