Subtyping sub-Saharan esophageal squamous cell carcinoma by comprehensive molecular analysis.
Esophageal squamous cell carcinoma (ESCC) is endemic in regions of sub-Saharan Africa (SSA), where it is the third most common cancer. Here, we describe whole-exome tumor/normal sequencing and RNA transcriptomic analysis of 59 patients with ESCC in Malawi. We observed similar genetic aberrations as reported in Asian and North American cohorts, including mutations of TP53, CDKN2A, NFE2L2, CHEK2, NOTCH1, FAT1, and FBXW7. Analyses for nonhuman sequences did not reveal evidence for infection with HPV or other occult pathogens. Mutational signature analysis revealed common signatures associated with aging, cytidine deaminase activity (APOBEC), and a third signature of unknown origin, but signatures of inhaled tobacco use, aflatoxin and mismatch repair were notably absent. Based on RNA expression analysis, ESCC could be divided into 3 distinct subtypes, which were distinguished by their expression of cell cycle and neural transcripts. This study demonstrates discrete subtypes of ESCC in SSA, and suggests that the endemic nature of this disease reflects exposure to a carcinogen other than tobacco and oncogenic viruses.


Promising strategy for developing mRNA-based universal influenza virus vaccine for human population, poultry, and pigs- focus on the bigger picture.
Since the first outbreak in the 19th century influenza virus has remained emergent owing to the huge pandemic potential. Only the pandemic of 1918 caused more deaths than any war in world history. Although two types of influenza- A (IAV) and B (IBV) cause epidemics annually, influenza A deserves more attention as its nature is much wilier. IAVs have a large animal reservoir and cause the infection manifestation not only in the human population but in poultry and domestic pigs as well. This many-sided characteristic of IAV along with the segmented genome gives rise to the antigenic drift and shift that allows evolving the new strains and new subtypes, respectively. As a result, the immune system of the body is unable to recognize them. Importantly, several highly pathogenic avian IAVs have already caused sporadic human infections with a high fatality rate (~60%). The current review discusses the promising strategy of using a potentially universal IAV mRNA vaccine based on conserved elements for humans, poultry, and pigs. This will better aid in averting the outbreaks in different susceptible species, thus, reduce the adverse impact on agriculture, and economics, and ultimately, prevent deadly pandemics in the human population.


The effectiveness of ventriculocystocisternostomy for suprasellar arachnoid cysts.
The aim of this study was to report the long-term outcomes of patients receiving endoscopic ventriculocystocisternostomy (VCC) for suprasellar arachnoid cysts (SACs), and to analyze all published reports on outcomes of ventriculocystostomy (VC) versus VCC to compare the effectiveness of the 2 techniques. Eleven consecutive patients with previously untreated SACs were surgically treated using endoscopic VCC. Another 2 patients were treated with VCC following ventriculoperitoneal shunt placement. Clinical imaging data were recorded. An analysis was performed of all published patient outcomes following endoscopic VC or VCC for an SAC. Developmental delay and progressive macrocephaly were the most common preoperative symptoms. At a mean clinical follow-up interval of 63 months, 10 of 11 patients undergoing primary VCC did not require reoperation. An analysis of the literature suggests that VCC may be more effective than VC. Of the 44 reported patients that underwent VC as a first treatment, 7 (16%) required reoperation, and 7 (8%) of 86 patients who underwent VCC as a first treatment required reoperation. When VC or VCC was performed following a prior surgical procedure, 4 of 11 patients undergoing VC had a treatment failure requiring reoperation. In contrast, only 2 of the 17 reported cases of VCC following a prior procedure required further treatment. The difference in reoperation rates following either primary or secondary VC was significantly higher than following primary or secondary VCC (p = 0.04). The authors conclude that VCC is an effective and durable treatment for symptomatic SACs in most cases.


Altered Mental Status: Current Evidence-based Recommendations for Prehospital Care.
In the United States emergency medical services (EMS) protocols vary widely across jurisdictions. We sought to develop evidence-based recommendations for the prehospital evaluation and treatment of a patient with an acute change in mental status and to compare these recommendations against the current protocols used by the 33 EMS agencies in the State of California. We performed a literature review of the current evidence in the prehospital treatment of a patient with altered mental status (AMS) and augmented this review with guidelines from various national and international societies to create our evidence-based recommendations. We then compared the AMS protocols of each of the 33 EMS agencies for consistency with these recommendations. The specific protocol components that we analyzed were patient assessment, point-of-care tests, supplemental oxygen, use of standardized scoring, evaluating for causes of AMS, blood glucose evaluation, toxicological treatment, and pediatric evaluation and management. Protocols across 33 EMS agencies in California varied widely. All protocols call for a blood glucose check, 21 (64%) suggest treating adults at <60mg/dL, and half allow for the use of dextrose 10%. All the protocols recommend naloxone for signs of opioid overdose, but only 13 (39%) give specific parameters. Half the agencies (52%) recommend considering other toxicological causes of AMS, often by using the mnemonic AEIOU TIPS. Eight (24%) recommend a 12-lead electrocardiogram; others simply suggest cardiac monitoring. Fourteen (42%) advise supplemental oxygen as needed; only seven (21%) give specific parameters. In terms of considering various etiologies of AMS, 25 (76%) give instructions to consider trauma, 20 (61%) to consider stroke, and 18 (55%) to consider seizure. Twenty-three (70%) of the agencies have separate pediatric AMS protocols; others include pediatric considerations within the adult protocol. Protocols for patients with AMS vary widely across the State of California. The evidence-based recommendations that we present for the prehospital diagnosis and treatment of this condition may be useful for EMS medical directors tasked with creating and revising these protocols.


Clinical and experimental applications of NIR-LED photobiomodulation.
This review presents current research on the use of far-red to near-infrared (NIR) light treatment in various in vitro and in vivo models. Low-intensity light therapy, commonly referred to as "photobiomodulation," uses light in the far-red to near-infrared region of the spectrum (630-1000 nm) and modulates numerous cellular functions. Positive effects of NIR-light-emitting diode (LED) light treatment include acceleration of wound healing, improved recovery from ischemic injury of the heart, and attenuated degeneration of injured optic nerves by improving mitochondrial energy metabolism and production. Various in vitro and in vivo models of mitochondrial dysfunction were treated with a variety of wavelengths of NIR-LED light. These studies were performed to determine the effect of NIR-LED light treatment on physiologic and pathologic processes. NIRLED light treatment stimulates the photoacceptor cytochrome c oxidase, resulting in increased energy metabolism and production. NIR-LED light treatment accelerates wound healing in ischemic rat and murine diabetic wound healing models, attenuates the retinotoxic effects of methanol-derived formic acid in rat models, and attenuates the developmental toxicity of dioxin in chicken embryos. Furthermore, NIR-LED light treatment prevents the development of oral mucositis in pediatric bone marrow transplant patients. The experimental results demonstrate that NIR-LED light treatment stimulates mitochondrial oxidative metabolism in vitro, and accelerates cell and tissue repair in vivo. NIR-LED light represents a novel, noninvasive, therapeutic intervention for the treatment of numerous diseases linked to mitochondrial dysfunction.


Anthropometric measures in cardiovascular disease prediction: comparison of laboratory-based versus non-laboratory-based model.
Body mass index (BMI) has been used to simplify cardiovascular risk prediction models by substituting total cholesterol and high-density lipoprotein cholesterol. In the elderly, the ability of BMI as a predictor of cardiovascular disease (CVD) declines. We aimed to find the most predictive anthropometric measure for CVD risk to construct a non-laboratory-based model and to compare it with the model including laboratory measurements. The study included 2675 women and 1902 men aged 55-79 years from the prospective population-based Rotterdam Study. We used Cox proportional hazard regression analysis to evaluate the association of BMI, waist circumference, waist-to-hip ratio and a body shape index (ABSI) with CVD, including coronary heart disease and stroke. The performance of the laboratory-based and non-laboratory-based models was evaluated by studying the discrimination, calibration, correlation and risk agreement. Among men, ABSI was the most informative measure associated with CVD, therefore ABSI was used to construct the non-laboratory-based model. Discrimination of the non-laboratory-based model was not different than laboratory-based model (c-statistic: 0.680-vs-0.683, p=0.71); both models were well calibrated (15.3% observed CVD risk vs 16.9% and 17.0% predicted CVD risks by the non-laboratory-based and laboratory-based models, respectively) and Spearman rank correlation and the agreement between non-laboratory-based and laboratory-based models were 0.89 and 91.7%, respectively. Among women, none of the anthropometric measures were independently associated with CVD. Among middle-aged and elderly where the ability of BMI to predict CVD declines, the non-laboratory-based model, based on ABSI, could predict CVD risk as accurately as the laboratory-based model among men.


Obesity, diet, endogenous estrogens, and the risk of hormone-sensitive cancer.
In postmenopausal women, obesity is positively related to the risk of both breast and endometrial cancer. Additionally, obesity is associated with increased estrogen production secondary to increased peripheral aromatization. In postmenopausal women, this effect is proportionately more significant because the ovaries no longer contribute to production of estrogen. Obesity also alters the further metabolism of estrogens, resulting in products that retain estrogenic potency. Consequently, the estrogen-sensitive tissues of obese women are exposed to more stimulation than those tissues in leaner women. It is possible that this increased estrogen exposure is responsible for the augmented risk of breast and endometrial cancer observed in obesity. Evidence is accumulating that dietary macronutrients can alter the metabolism and excretion of endogenous estrogen and androgen, providing a potential mechanism by which diet can modulate the risk of hormone-sensitive cancers.


Regression models for relative survival.
Four approaches to estimating a regression model for relative survival using the method of maximum likelihood are described and compared. The underlying model is an additive hazards model where the total hazard is written as the sum of the known baseline hazard and the excess hazard associated with a diagnosis of cancer. The excess hazards are assumed to be constant within pre-specified bands of follow-up. The likelihood can be maximized directly or in the framework of generalized linear models. Minor differences exist due to, for example, the way the data are presented (individual, aggregated or grouped), and in some assumptions (e.g. distributional assumptions). The four approaches are applied to two real data sets and produce very similar estimates even when the assumption of proportional excess hazards is violated. The choice of approach to use in practice can, therefore, be guided by ease of use and availability of software. We recommend using a generalized linear model with a Poisson error structure based on collapsed data using exact survival times. The model can be estimated in any software package that estimates GLMs with user-defined link functions (including SAS, Stata, S-plus, and R) and utilizes the theory of generalized linear models for assessing goodness-of-fit and studying regression diagnostics.


Total hip arthroplasty for the treatment of ankylosed hips : a five to twenty-one-year follow-up study.
The purpose of the present retrospective study was to report the long-term results of total hip arthroplasty following a hip fusion. Special attention was paid to the resulting function of both the involved joint and the neighboring joints, as pain in the lower back or knee was the usual indication for conversion. The factors that were likely to influence the functional outcome were analyzed. Forty-five consecutive total hip arthroplasties were performed in forty-five patients from 1969 through 1993. The mean age of the patients at the time of the operation was 55.8 years (range, twenty-eight to eighty years). Ankylosis of the hip had been spontaneous in twenty patients and postoperative in twenty-five patients. The mean duration of the ankylosis had been thirty-six years (range, three to sixty-five years). The mean duration of follow-up was 8.5 years (range, five to twenty-one years). No patient was lost to follow-up. The mean hip score, according to the scale of Merle d'Aubigné, was 16.5 1.5 points at the latest follow-up evaluation. Hip function was considered to be satisfactory for forty-one (91%) of the forty-five patients. The definitive score for walking ability was not achieved by the one-year evaluation; it improved notably for two to three years and then it remained stable. At the time of the latest follow-up, the mean arc of flexion was 88 degrees (range, 30 degrees to 130 degrees ). Forty-three (96%) of the forty-five patients had no pain in the involved joint. The only factor that was predictive of the final functional result with regard to walking ability was the intraoperative status of the gluteal muscles. Most patients had effective pain relief in the neighboring joints. The cumulative survival rate at eight years, with revision as the end point, was 96.7% (95% confidence interval, 90.2% to 100%). The long-term effectiveness of total hip arthroplasty for the treatment of an ankylosed hip was clearly demonstrated in both the involved and the neighboring joints in the present study. However, the preoperative and intraoperative status of the gluteal muscles should be carefully evaluated when this procedure is being considered, as this was the only factor that was predictive of the final walking ability.


[In vivo effect of rhG-CSF on the CXCR-4 expression of hematopoietic progenitor or stem cells in bone marrow and peripheral blood].
To investigate the effect of in vivo rhG-CSF on the CXCR-4 expression of hematopoietic progenitor or stem cells in bone marrow and peripheral blood, the expressions of CXCR-4 on CD34(+) cells and mononuclear cells of bone marrow and peripheral blood from healthy donor before and after mobilization were detected by three-color fluorescence analysis. The results showed that a significantly higher expression of CXCR4 on CD34(+) cells of bone marrow and mononuclear cells of peripheral blood, as compared to those before mobilization. There were no significant differences of CXCR-4 expression of CD34(+) cells in peripheral blood after mobilization, as compared with steady state bone marrow, and no dynamic change of mononuclear cells expressing CXCR-4 in bone marrow before and after mobilization. Significant positive correlation were found between the percentage of CD34(+) cells in bone marrow before mobilization and that in bone marrow and peripheral blood after mobilization; furthermore, the percentage of CD34(+) cells of bone marrow before mobilization had a positive correlation with both the count of CD34(+) cells per kilogram on the day of collection in bone marrow and peripheral blood after mobilization. It is concluded that the mobilization of hematopoietic cells may be involved in the signaling of SDF-1/CXCR-4 according to the increase of the surface expression of CXCR-4 on CD34(+) cells in bone marrow and on the MNC in peripheral blood after mobilization; meanwhile, the high surface expression of CXCR-4 may contribute to the MNC engraftment, monitoring the percentage of CD34(+) cells in bone marrow before mobilization can be regarded as a predictive factor for mobilization outcome.


Donor spermatogenesis in de novo formed seminiferous tubules from transplanted testicular cells in rhesus monkey testis.
Can transplanted primate testicular cells form seminiferous tubules de novo, supporting complete spermatogenesis? Cryopreserved testicular cells from a prepubertal monkey can reorganize in an adult monkey recipient testis forming de novo seminiferous tubular cords supporting complete spermatogenesis. De novo morphogenesis of testicular tissue using aggregated cells from non-primate species grafted either subcutaneously or in the testis can support spermatogenesis. Two postpubertal rhesus monkeys (Macaca mulatta) were given testicular irradiation. One monkey was given GnRH-antagonist treatment from 8 to 16 weeks after irradiation, while the other received sham injections. At 16 weeks, cryopreserved testicular cells from two different prepubertal monkeys [43 × 106 viable (Trypan-blue excluding) cells in 260 μl, and 80 × 106 viable cells in 400 μl] were transplanted via ultrasound-guided injections to one of the rete testis in each recipient, and immune suppression was given. The contralateral testis was sham transplanted. Testes were analyzed 9 months after transplantation. Spermatogenic recovery was assessed by testicular volume, weight, histology and immunofluorescence. Microsatellite genotyping of regions of testicular sections obtained by LCM determined whether the cells were derived from the host or transplanted cells. Transplanted testis of the GnRH-antagonist-treated recipient, but not the sham-treated recipient, contained numerous irregularly shaped seminiferous tubular cords, 89% of which had differentiating germ cells, including sperm in a few of them. The percentages of donor genotype in different regions of this testis were as follows: normal tubule, 0%; inflammatory, 0%; abnormal tubule region, 67%; whole interior of abnormal tubules, >99%; adluminal region of the abnormal tubules, 92%. Thus, these abnormal tubules, including the enclosed germ cells, were derived de novo from the donor testicular cells. Not applicable. The de novo tubules were observed in only one out of the two monkeys transplanted with prepubertal donor testicular cells. These findings may represent a promising strategy for restoration of fertility in male childhood cancer survivors. The approach could be particularly useful in those exposed to therapeutic agents that are detrimental to the normal development of the tubule somatic cells affecting the ability of the endogenous tubules to support spermatogenesis, even from transplanted spermatogonial stem cells. This work was supported by research grants P01 HD075795 from Eunice Kennedy Shriver National Institute of Child Health & Human Development (NICHD/NIH) to K.E.O and Cancer Center Support Grant P30 CA016672 from NCI/NIH to The University of Texas MD Anderson Cancer Center. The authors declare that they have no competing interests.


Early detection of salmonid alphavirus in seawater from marine farm sites of Atlantic salmon Salmo salar.
The traditional strategy for national surveillance of salmonid alphavirus (SAV) infection in Norwegian fish farms relies on a costly, time-consuming, and resource-demanding approach based on the monthly sampling of fish from all marine farms with salmonids. In order to develop an alternative surveillance method, a water filtration method was tested in parallel with the ongoing surveillance program at 7 Norwegian marine farm sites of Atlantic salmon Salmo salar L. with no current suspicion of SAV infection. During the period from May 2019 to January 2020, seawater samples were collected from the top layer water inside all net-pens at these 7 sites. The samples were concentrated for SAV by filtration through an MF-Millipore™ electronegative membrane filter, followed by rinsing with NucliSENS® Lysis Buffer, before RNA extraction and analysis by RT-qPCR. SAV was detected from seawater at an earlier stage compared to traditional sampling methods, at all sites where the fish tested positive for SAV. A significant negative relationship was observed at all sites between the SAV concentration found in seawater samples and the number of days until SAV was detected in the fish. This means that the fewer the SAV particles in the seawater, the more days it took until SAV was detected in the fish samples. Based on this, sampling of seawater every month for the surveillance of SAV has a great potential as an alternative method for early detection of SAV in Atlantic salmon farms.


Electrolyte Derangements, Hyperlactatemia, and Cardiac Abnormalities Secondary to Refeeding in Three Dogs: Case Report.
Three dogs that presented to the emergency service in severely emaciated body conditions were admitted to the hospital for monitoring and refeeding. During their hospitalization, all three dogs developed electrolyte derangements or required supplementation to prevent hypophosphatemia and hypomagnesemia. Additionally, all dogs developed hyperlactatemia, which was suspected to be secondary to thiamine deficiency. Two dogs were reported to have cardiac abnormalities, including cardiac arrhythmias, systolic dysfunction, and spontaneous echogenic contrast. These cases highlight the complexity of refeeding syndrome and its associated complications that extend beyond electrolyte deficiencies.


Milestones in DRI Development: What Does the Future Hold?
The state of nutritional health in the United States in the early part of the twentieth century was very different from today. Nutrient deficiencies and dental caries were prevalent health concerns for many Americans. In 1940, the US National Defense Advisory Commission asked the National Academy of Sciences for help in studying problems of nutrition in the United States. The outcome was issuance of the first RDAs. The goal of the RDAs was to recommend "…allowances sufficiently liberal to be suitable for maintenance of good nutritional status." In the subsequent decades, a very different nutritional health challenge began to emerge for an increasing proportion of the population, that of overweight and obesity and risk of diet-related chronic disease. In part, as a response to this challenge, the RDA process was revised and the Dietary Reference Intakes (DRIs) were developed. The DRIs are a set of reference values that, when adhered to, predict a low probability of nutrient inadequacy or excessive intake. Recently, new DRI guidelines were proposed to define reference points for nutrient and food component intakes that influence risk of chronic disease. Developing DRIs for chronic disease endpoints presents unique challenges, notably, chronic diseases are multifactorial in nature and not directly nutrient-specific; the body of evidence supporting nutrients and other food substances as modifiers of risk of chronic disease is generally limited; and there is a lack of consistency in findings across study types. In addition, the latency of dietary exposures and chronic disease outcomes makes it difficult to demonstrate causality. Adapting the DRI model to meet the needs of the general population in the current context suggests a need to redefine the boundaries that describe the health of the population and to re-examine how indicators of chronic disease can be integrated effectively into the DRI process.


[Laparoscopic cholecystectomy in Spain: multicentric study of 2,432 patients].
To evaluate the efficiency of laparoscopic cholecystectomy for the treatment of gallstones. Retrospective/prospective multicentric study undertaken by the Department of Surgical Endoscopy of the Spanish Association of Surgeons. Data were introduced into a computerized protocol. Statistical analysis with SPSS 4.0 applying the Student's T test and Pearson's chi square. 69 surgeons from 40 Hospitals who have performed laparoscopic cholecystectomies in 2432 patients, diagnosed of gallstones, with an average age of 49 years (9-91) and 80% in women. The operation was completed in 94% of the cases and lasted on an average of 75 minutes (20-120). Surgical findings were 2380 non complicated gallstones, 185 acute cholecystitis (7.6%) and 265 scleroatrophic gallbladders (11%). Intraoperative cholangiography was performed in 9% and gallstones in the common bile duct were diagnosed in 2%, of which 0.3% were treated by laparoscopy and 1% by post-operative endoscopic sphinterotomy Five per cent were converted to laparotomy, of which 29% (1.5% of total patients) were obligatory conversions. Among postoperative complications there were 0.4% lesions due to abdominal puncture, 0.4% lesions of the common bile duct and 2.5% hemorrhages. Post-operative complications were seen in 7.5% of patients. The most frequent complications affected the surgical wounds (3%), whereas bile duct complications represented 0.6% and were most frequent in the series of less than 50 patients. Mortality was 0.12%. Average post-operative hospital stay was 2.5 days (0.5-5). In Spain, laparoscopic cholecystectomy is an effective and safe procedure with results which are comparable to other multicentric studies published in other countries, with a morbidity-morbility index similar to open cholecystectomy and the great added advantage is the avoidance of laparotomy and its inconveniences (pain, longer hospital stay...) and complications.


Management of a severe forceful breather with Rett syndrome using carbogen.
We have used a novel neurophysiological technique in the NeuroScope system in combination with conventional electroencephalography (EEG) to monitor both brainstem and cortical activity simultaneously in real-time in a girl with Rett syndrome. The presenting clinical features in our patient were severe sleep disturbances, irregular breathing in the awake state dominated by Valsalva's type of breathing followed by tachypnoea and very frequent attacks of seizures and vacant spells. Our novel neurophysiological data showed that the patient was a Forceful Breather according to the breathing categories in Rett syndrome. She had frequent abnormal spontaneous brainstem activation (ASBA) preceded by severe attacks of hypocapnoea, which was caused by a combination of Valsalva's type of breathing and tachypnoea and all these together were responsible for the seizures and non-epileptic vacant spells. The ASBA was not detectable in conventional EEG and there were no epileptiform changes in the EEG during the seizures and vacant spells caused by the hypocapnic attacks, therefore these were pseudo-seizures. The record of brainstem activity confirmed that these were autonomic events, a kind of "brainstem epilepsy". We successfully treated the sleep disturbance with Pipamperone, a 5-hydroxytryptophan antagonist of receptor type 2 and we prevented the severe hypocapnoea during Valsalva's type of breathing and during tachypnoea using carbogen (a mixture of 5% carbon dioxide and 95% oxygen), which we gave by inhalation. Our treatment drastically reduced the autonomic events, promoted whole night sleep and significantly improved the quality of life in our patient. She can now participate in normal family activity which was previously impossible before treatment.


Anomalously Low Barrier for Water Dimer Diffusion on Cu(111).
A molecular-scale description of water and ice is important in fields as diverse as atmospheric chemistry, astrophysics, and biology. Despite a detailed understanding of water and ice structures on a multitude of surfaces, relatively little is known about the kinetics of water motion on surfaces. Here, we report a detailed study on the diffusion of water monomers and the formation and diffusion of water dimers through a combination of time-lapse low-temperature scanning tunnelling microscopy experiments and first-principles electronic structure calculations on the atomically flat Cu(111) surface. On the basis of an unprecedented long-time study of individual water monomers and dimers over days, we establish rates and mechanisms of water monomer and dimer diffusion. Interestingly, we find that the monomer and the dimer diffusion barriers are similar, despite the significantly larger adsorption energy of the dimer. This is thus a violation of the rule of thumb that relates diffusion barriers to adsorption energies, an effect that arises because of the directional and flexible hydrogen bond within the dimer. This flexibility during diffusion should also be relevant for larger water clusters and other hydrogen-bonded adsorbates. Our study stresses that a molecular-scale understanding of the initial stages of ice nanocluster formation is not possible on the basis of static structure investigations alone.


Criminal networks analysis in missing data scenarios through graph distances.
Data collected in criminal investigations may suffer from issues like: (i) incompleteness, due to the covert nature of criminal organizations; (ii) incorrectness, caused by either unintentional data collection errors or intentional deception by criminals; (iii) inconsistency, when the same information is collected into law enforcement databases multiple times, or in different formats. In this paper we analyze nine real criminal networks of different nature (i.e., Mafia networks, criminal street gangs and terrorist organizations) in order to quantify the impact of incomplete data, and to determine which network type is most affected by it. The networks are firstly pruned using two specific methods: (i) random edge removal, simulating the scenario in which the Law Enforcement Agencies fail to intercept some calls, or to spot sporadic meetings among suspects; (ii) node removal, modeling the situation in which some suspects cannot be intercepted or investigated. Finally we compute spectral distances (i.e., Adjacency, Laplacian and normalized Laplacian Spectral Distances) and matrix distances (i.e., Root Euclidean Distance) between the complete and pruned networks, which we compare using statistical analysis. Our investigation identifies two main features: first, the overall understanding of the criminal networks remains high even with incomplete data on criminal interactions (i.e., when 10% of edges are removed); second, removing even a small fraction of suspects not investigated (i.e., 2% of nodes are removed) may lead to significant misinterpretation of the overall network.


Pannexin1 and pannexin3 delivery, cell surface dynamics, and cytoskeletal interactions.
Pannexins (Panx) are a class of integral membrane proteins that have been proposed to exhibit characteristics similar to those of connexin family members. In this study, we utilized Cx43-positive BICR-M1R(k) cells to stably express Panx1, Panx3, or Panx1-green fluorescent protein (GFP) to assess their trafficking, cell surface dynamics, and interplay with the cytoskeletal network. Expression of a Sar1 dominant negative mutant revealed that endoplasmic reticulum to Golgi transport of Panx1 and Panx3 was mediated via COPII-dependent vesicles. Distinct from Cx43-GFP, fluorescence recovery after photobleaching studies revealed that both Panx1-GFP and Panx3-GFP remained highly mobile at the cell surface. Unlike Cx43, Panx1-GFP exhibited no detectable interrelationship with microtubules. Conversely, cytochalasin B-induced disruption of microfilaments caused a severe loss of cell surface Panx1-GFP, a reduction in the recoverable fraction of Panx1-GFP that remained at the cell surface, and a decrease in Panx1-GFP vesicular transport. Furthermore, co-immunoprecipitation and co-sedimentation assays revealed actin as a novel binding partner of Panx1. Collectively, we conclude that although Panx1 and Panx3 share a common endoplasmic reticulum to Golgi secretory pathway to Cx43, their ultimate cell surface residency appears to be independent of cell contacts and the need for intact microtubules. Importantly, Panx1 has an interaction with actin microfilaments that regulates its cell surface localization and mobility.


The relation between experiential avoidance, alexithymia and emotion regulation in inpatient adolescents.
Recently, efforts have been made to better understand constructs that are associated with difficulties in emotion regulation in hopes of identifying underlying mechanisms that may be valuable targets for intervention. Against this background, the present study had two aims. Firstly, we wanted to explore the relation between emotion regulation, experiential avoidance and alexithymia by determining whether adolescents with elevated scores on a measure of alexithymia would report deficits in emotion regulation and experiential avoidance. Secondly, we sought to evaluate the role of experiential avoidance as a mediator in the relation between alexithymia and emotion regulation. The sample (N = 64) consisted of adolescents recruited from an inpatient facility of which approximately 30% were classified as having alexithymia. The results of this study indicate that adolescents with alexithymia report deficits in emotion regulation and elevated experiential avoidance. Experiential avoidance mediated the relation between alexithymia and emotion regulation, indicating that while the inability to effectively use language to identify and describe emotional states is strongly correlated with difficulties in regulating one's emotions, this relation is mediated by the unwillingness to tolerate aversive private experiences. Limitations and strengths of the present study are also noted.


The effects of delayed diagnosis and treatment in patients with an occult spinal dysraphism.
From 1987 to 1993, 21 older individuals presented for the first time with signs and symptoms that eventually led to the diagnosis of occult spinal dysraphism. Assessment consisted of a neurological examination, urodynamic studies preoperatively and postoperatively, and spinal cord imaging. Of 21 patients 18 had an abnormal neurological examination, whereas only 15 had an abnormal urodynamic study, as judged by sphincter electromyography. Radiological imaging showed that 9 patients had a tethered cord alone, 4 each had lipomeningocele and lipoma, 2 had a bony spine abnormality and 1 each had thoracic meningocele and diastematomyelia. Of the 21 patients 19 underwent spinal surgery. Postoperatively, the neurological examination improved in 1 case (5%) and remained unchanged in 18 (95%), while urodynamic findings improved in 3 (16%), were unchanged in 11 (68%) and worsened in 5 (26%). Six patients had progressive deterioration and required secondary spinal surgery, which helped only 2 (33%). These observations confirm that older children and adults with occult spinal dysraphism are more likely to present with irreversible urological and neurological findings than younger children, and so it is imperative that a diagnosis be made and treatment be instituted as early as possible.


C-CAT: The National Datacenter for Cancer Genomic Medicine in Japan.
Since June 2019, under the umbrella of the national health insurance system, Japan has started cancer genomic medicine (CGM) with comprehensive genomic profiling (CGP) tests. The Ministry of Health, Labour and Welfare (MHLW) of Japan constructed a network of CGM hospitals (a total of 233 institutes as of July 1, 2022) and established the Center for Cancer Genomics and Advanced Therapeutics (C-CAT), the national datacenter for CGM. Clinical information and genomic data from the CGP tests are securely transferred to C-CAT, which then generates "C-CAT Findings" reports containing information of clinical annotation and matched clinical trials based on the CGP data. As of June 30, 2022, a total of 36,340 datapoints of clinical/genomic information are aggregated in C-CAT, and the number is expected to increase swiftly. The data are now open for sharing with not only the CGM hospitals but also other academic institutions and industries.


[Use of local hyperthermia in treating cancer patients].
Basic research in thermobiology made it possible to substantiate the advisability of incorporation of local hyperthermia in schemes of radiotherapy of patients with tumors of different sites. Positive experience of thermoradiotherapy of 301 patients with laryngeal, head and neck, uterine and rectal tumors and osteogenic sarcoma using the Soviet equipment indicated the advisability of wider employment of local hyperthermia in various schemes of radio- and combined therapy.


p53 amyloid formation leading to its loss of function: implications in cancer pathogenesis.
The transcriptional regulator p53 has an essential role in tumor suppression. Almost 50% of human cancers are associated with the loss of p53 functions, where p53 often accumulates in the nucleus as well as in cytoplasm. Although it has been previously suggested that amyloid formation could be a cause of p53 loss-of-function in subset of tumors, the characterization of these amyloids and its structure-function relationship is not yet established. In the current study, we provide several evidences for the presence of p53 amyloid formation (in human and animal cancer tissues); along with its isolation from human cancer tissues and the biophysical characterization of these tissue-derived fibrils. Using amyloid seed of p53 fragment (P8, p53(250-257)), we show that p53 amyloid formation in cells not only leads to its functional inactivation but also transforms it into an oncoprotein. The in vitro studies further show that cancer-associated mutation destabilizes the fold of p53 core domain and also accelerates the aggregation and amyloid formation by this protein. Furthermore, we also show evidence of prion-like cell-to-cell transmission of different p53 amyloid species including full-length p53, which is induced by internalized P8 fibrils. The present study suggests that p53 amyloid formation could be one of the possible cause of p53 loss of function and therefore, inhibiting p53 amyloidogenesis could restore p53 tumor suppressor functions.


[Psychiatric disorders in a rural area: descriptive study of 2106 cases].
INTROD: Clinical samples gives a piece of information mediated by singular characteristics of the area (general practitioners, health services organization, needs of care feld by family or patient), but they are an important way in the study of psychosocial variables and their influence in pathways and morbidity of psychiatric disorders. MET: We obtained by SISMA survey (Information System for Mental Health in Andalucia) clinical and sociodemographic variables of a 2,016 Mental Health Center sample consulting during 40 months (attendance people = 90,368, rural area). We realized a descriptive and association study, with special attention to gender as modulating another factors related with health services use, diagnostic and clinical features. RES: ICD-10 Diagnostic groups of our sample was Affective disorders (F.3x) (36.5%, OR 1.99 for female), Neurotic disorders (F.4x) (29.75%, OR 2.01 for female) and Schizophrenia and related disorders (F.2x) (9.83%, OR 0.31 for female). The female-male ratio of our clinical sample was 1.70, incremented to 2.31 in geriatrics. Woman significantly consulted by themselves and men by their family. Married marital status was a help factor of psychopathology in women, as single status in men. Work activity and high or medium educative level was protective factors of psychopathology. CONCLUS: Clinical and sociodemographic variables we have seen, give us similar results to another spanish samples, too similar to actually accepted in Psychiatric Epidemiology. Female gender and class-linked vulnerability have an important influence as help-factors for specialized psychiatric treatment in our community.


The effects of environmental enrichment on hatchery-performance, smolt migration and capture rates in landlocked Atlantic salmon.
Enrichment of rearing environment with natural elements has been suggested to improve the welfare and post-release survival of cultured fish. We studied the combined effects of shelter structures, periodical water flow and water level changes on pre- and post-release performance of critically endangered landlocked Atlantic salmon (Salmo salar m. sebago). Relative to standard (plain) rearing tanks, provision of enrichment improved fish condition factor and survival during the first year of rearing when most mortality was attributable to parasitic and bacterial infections. The consequent higher density in enriched tanks probably induced greater growth variation and more dorsal fin damages than found in fish of standard tanks. Possibly this was partly due to the applied changes in water level. Experimentally determined smolt migration tendency at age 3 did not differ, on average, between the rearing groups, but enriched-reared fish showed clearly less variation in total movement activity than standard-reared fish. Experimental angling in earthen ponds did not suggest divergent vulnerability between the differentially reared fish at age 3, but decreased condition during the preceding growth season increased vulnerability to fishing. Based on long-term post-stocking tag returns in large-lake fisheries, fish length at release but not rearing method affected the capture rates of fish released at age 2. When released at age 3 the fish grown in enriched environment had a higher risk to be captured with stationary gears and earlier by hook and line gears compared to standard-reared conspecifics. Earlier time of maximal smolt migration activity was associated with an increased risk of being captured. We suggest that environmental enrichment may modulate growth- and behavior-related qualities that indirectly increased the vulnerability to fishing in natural conditions but not in experimental setting. The favorable effects of enrichment on early survival encourages adopting enriched rearing practices in supportive breeding of landlocked salmon.


Biomarkers of Exposure among USA Adult Hookah Users: Results from Wave 1 of the Population Assessment of Tobacco and Health (PATH) Study (2013-2014).
Hookah smoking has become common in the USA, especially among young adults. This study measured biomarkers of exposure to known tobacco product toxicants in a population-based sample of exclusive, established hookah users. Urinary biomarker data from 1753 adults in Wave 1 of the Population Assessment of Tobacco and Health (PATH) Study were used to compare geometric mean concentrations of biomarkers of exposure in exclusive, established past 30-day hookah users to never users of tobacco. Geometric mean ratios were calculated comparing hookah user groups with never users adjusting for age, sex, race/ethnicity, education, past 30-day marijuana use, secondhand smoke exposure and creatinine. Past 30-day hookah users (n = 98) had 10.6 times the urinary cotinine level of never tobacco users. Compared to never tobacco users, past 30-day hookah users had 2.3 times the level of the carcinogen 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL), a metabolite of the tobacco-specific nitrosamine (TSNA) 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone (NNK), 1.3 times higher polycyclic aromatic hydrocarbons (PAHs) 3-hydroxyfluorene and 1-hydroxypyrene, 1.8 times higher levels of acrylonitrile, 1.3 times higher levels of acrylamide, and 1.2 times higher levels of acrolein exposure. These data indicate that hookah use is a significant source of exposure to nicotine, carcinogens, and respiratory toxicants.


Intraoperative Seizure Detection During Active Resection of Glioblastoma Through a Novel Hollow Circular Electrocorticography Array.
Data supporting the use of electrocorticography (ECoG) monitoring during electrical stimulation in awake craniotomies for resection of supratentorial neoplasms is robust, but its applicability during active resection is often limited by the inability to keep the array in place. Given the known survival benefit of gross total resection in glioma surgery, novel approaches to surgical monitoring are warranted to maximize safe resection and optimize surgical outcomes in patients with glioblastoma. A 68-yr-old right-handed woman presented to the emergency department with confusion. Imaging studies revealed a bifrontal intra-axial brain lesion. She underwent a left-sided awake craniotomy procedure with cortical and subcortical mapping. During surgical resection, multiple electrographic seizures were detected on continuous ECoG monitoring with a customized 22-channel high-density hollow circular array. She remained without clinical evidence of seizures at 3 mo after surgery. We report a unique case of serial electrographic seizures detected during continuous intraoperative ECoG monitoring during active surgical resection of a glioblastoma using a novel circular hollow array during an awake craniotomy. The use of continuous ECoG monitoring during active resection may provide additional data, with potential influence in outcomes for patients undergoing resection of high-grade glial neoplasms.


[Elaboration of a conditioned reflex in chronic alcoholics using an unrecognizable motivationally significant word].
20 patients suffering from chronic alcoholism and 10 healthy, righthanded persons with normal sight were examined. Alcoholics were trained to elaborate a temporary connection by means of unrecognized significant word "vodka" related to their dominant motivation. The amplitude of the late positive response P300 was greater and its latency was shorter comparing with responses to neutral words. The latency of P300 potential to a conditioned stimulus combined with the word "vodka" is reduced in all recorded areas, especially in the left occipital zone. Healthy persons did not manifest this facilitation. The amplitude of the potential considerably decreased in all recorded areas (occipital, parieto-temporo-occipital and central) especially in the left hemisphere. Conditioned changes of the P300 wave during conditioning by means of the unrecognized stimulus are stable and do not extinct despite multiple signal presentations without pairing with the significant word. The formation of conditioned connections by means of unrecognized stimulus in patients suffering from chronic alcoholism suggests that a certain role in psychical dependence upon alcohol may be played by unrecognized stimuli.


Monitoring mRNA transcription of genes involved in early pregnancy from endometrium and peripheral blood mononuclear cells of pregnant pigs with different parity.
The reproductive success of mammals is largely dependent on the interaction between maternal and foetal interfaces during early pregnancy. Particularly, immune cells which reside at the maternal endometrium can modulate the conception and placental vascularization. In this study, we analysed the transcription of genes involved in early pregnancy from endometrium and peripheral blood mononuclear cells (PBMCs) of pregnant pigs with different parity. Briefly, three groups of female pigs were divided based on parity (0, 2 and 5) and each group was artificially inseminated. Within 30 days of gestation, the total RNA was isolated from the endometrium and PBMCs of sacrificed experimental pigs and the expression patterns of genes involved in early pregnancy were monitored by quantitative real-time RT-PCR. Results indicated absence of correlation between increased parity and the expression of vascular endothelial growth factor (VEGF) and hypoxia-inducible factor 1-α (HIF-1α) mRNA in endometrium among the groups of pigs analysed. Yet, the mRNA levels of Fas, Fas ligand (FasL) and tumour necrosis factor-α (TNF-α) in the endometrium of parity 5 sows were much higher than those of pregnant gilts (parity 0), and the mRNA ratios of both TNF-α:interleukin-4 (IL-4) and IFN-γ (interferon-γ):interleukin-10 (IL-10) in PBMCs of pregnant pigs were augmented with increasing parity. Furthermore, the mRNA levels of TNF-α and IFN-γ in PBMCs of pregnant pigs were inversely correlated with litter size. These combined results may demonstrate that increased parity of pregnant pigs leads to enhance Th1-prone immunity within the maternal-foetal interface during early pregnancy.


Fecal calprotectin in inflammatory bowel disease.
Inflammatory bowel disease (IBD) and irritable bowel syndrome share many symptoms. While irritable bowel syndrome is a functional bowel disorder for which no specific treatment is available, the range of effective therapies for IBD is evolving rapidly. Accurate diagnosis of IBD is therefore essential. Clinical assessment, together with various imaging modalities and endoscopy, has been the mainstay of diagnosis for many years. Fecal biomarkers of gastrointestinal inflammation have appeared in the past decade, of which calprotectin, a neutrophil cytosolic protein, has been studied the most. Crohn's disease and ulcerative colitis are chronic remitting and relapsing diseases, and objective assessment of disease activity and response to treatment are important. This review focuses on the use of fecal calprotectin measurements in the diagnosis and monitoring of patients with IBD.


[Surgical treatment of lid retraction in Graves' ophthalmopathy].
Surgical treatment of lid retraction in Graves' ophthalmopathy consisted of resection of Müller's muscle and myotomy of Müller's muscle combined with intraoperative guided disinsertion of the levator palpebrae superioris from the tarsus. Among 10 eyes of 9 patients treated by the authors, 9 eyes achieved good cosmetic effect and corneal exposure was prevented during a follow-up of 3-42 months.


The effects of polydextrose and xylitol on microbial community and activity in a 4-stage colon simulator.
This study focused on the effects of candidate prebiotics polydextrose (PDX) and xylitol on the microbial community and its metabolic activity in a colon simulator. A semicontinuous, anaerobic culture system was used with 4 vessels mimicking the conditions in the human large intestine from proximal to distal colon. Bacterial inocula for the independent simulations were obtained from fecal samples of different donors. Synthetic medium, mimicking the contents of the small intestine, containing either 2% of the prebiotic candidate or no added carbohydrates as a control, was fed to the system. After 48 h of simulation samples were collected and analyzed. A sustained degradation of polydextrose throughout the colon model and a more rapid degradation of xylitol were observed. The fermentation of both compounds was characterized by a significantly increased production of short-chain fatty acids (SCFA). Polydextrose increased the concentrations of all SCFA, especially acetate and propionate, and xylitol especially the concentration of butyrate. Branched-chain fatty acids (BCFA) levels decreased significantly as a result of polydextrose and xylitol supplementation, whereas biogenic amine levels remained mostly unchanged. Thus, a beneficial shift in the metabolic patterns of the colon microbes was measured with both of the tested products. These in vitro studies provide evidence to the prebiotic characteristics of polydextrose; also, further beneficial properties of xylitol were demonstrated in the colon model.


There's an App for That: Using Geosocial Networking Apps to Access Young Black Gay, Bisexual, and other MSM at Risk for HIV.
Young black gay, bisexual, and other MSM (YBMSM) that carry a disproportionate HIV burden in the US Geosocial networking applications (GSN apps) are environments that may increase HIV risk among users. This study explored the acceptability and feasibility of using these apps for HIV/sexually transmitted infection (STI) public health outreach. Semi-structured in-depth qualitative interviews. A frequently reported GSN app for meeting sex partners by newly diagnosed HIV-infected MSM in Baltimore. Seventeen YBMSM aged 18 to 24 (mean = 21.5/SD = 1.8) who were logged-on to the GSN app in venues or census tracts in high HIV transmission areas. Participants completed 60 to 90 minute semi-structured interviews, which were audio-recorded and transcribed. Interview data were analyzed in NVivo10 using categorical analysis and double-coded until consistency was achieved. Participants described GSN apps as acceptable and feasible resources for public health practitioners seeking to access YBMSM to provide HIV/STI treatment and prevention services and resources. Three themes emerged: (1) the need to authenticate public health messages to distinguish from spam; (2) improved access to YBMSM including opportunities to identify and access virtual congregations of youth in non-gay-related spaces; and (3) the importance of avoiding stigmatizing YBMSM when targeting sexual health messages. GSN apps have great potential as tools for identifying and engaging at-risk YBMSM. Additional work is needed to understand limitations of this medium, to develop strategies to engage YBMSM without further stigmatizing them, and to maximize their outreach potential.


Risk factors in body image dissatisfaction: gender, maladaptive perfectionism, and psychological wellbeing.
Discontent with one's appearance (body image dissatisfaction) has become a global phenomenon, associated with the development of extreme behaviours in order to correct perceived body image problems. Much of the literature has focused on associated destructive behaviours that result from body image dissatisfaction; however, there has been a dearth of research examining risk factors for body image problems in adults. Thus, the current study aimed to investigate risk factors associated with adult body image problems, focusing on two variables highlighted in the literature (gender and maladaptive perfectionism). The current study (1) compared gender differences in body image dissatisfaction and (2) investigated psychological wellbeing as a mediator between maladaptive perfectionism and body image dissatisfaction. The sample included 139 Australian adults from universities (55.7%) and the community (44.3%). Participants completed demographic questions, the Frost Multidimensional Perfectionism Scale, the Psychological Wellbeing questionnaire, the Multidimensional Body-Self Relations Questionnaire, and Marlowe-Crowne Social Desirability Scale. In contrast to predictions, females had higher body area satisfaction and appearance evaluation scores. Furthermore, in line with the hypotheses, psychological wellbeing fully mediated the relationship between maladaptive perfectionism and poor body image. The findings suggest gender, maladaptive perfectionism, and psychological wellbeing may act as risk factors for body image dissatisfaction.


Digital subtraction arthrography of the wrist.
Digital subtraction arthrography of the wrist was used to identify abnormalities in eighty-six (60 per cent) of 139 patients during a fifteen-month period. Multiple abnormalities were noted in thirty-four (25 per cent) of the wrists. The clinical signs and symptoms in the eighty-six wrists did not always correlate with the defects that were seen on the arthrograms. Three of five patients who had an isolated tear of the scapholunate ligament, six of thirteen who had an isolated tear of the lunotriquetral ligament, and seven of nineteen who had an isolated tear of the triangular fibrocartilage complex also had signs and symptoms on the opposite side of the wrist. Many of the lesions that were seen on arthrography may have been serendipitous, degenerative, or unrelated to a specific injury. There was a high prevalence of positive ulnar variance in patients who had at least one ulnar abnormality. Capsular tears, most often seen on the radiovolar aspect of the wrist, were best outlined by contrast medium injected into the radiocarpal joint. The arthroscopic findings differed from the arthrographic findings in five of the twenty patients in whom both studies were done. The three-compartment technique of injection is a valuable diagnostic tool. Injections of contrast medium into the distal radio-ulnar joint outlined five of thirteen tears of the triangular fibrocartilage complex that were not seen after injection into the radiocarpal joint. Of the eleven tears that were seen after injection into the radiocarpal joint, five were not seen when contrast medium was injected into the distal radio-ulnar joint.


Drosophila melanogaster: a case study of a model genomic sequence and its consequences.
The sequencing and annotation of the Drosophila melanogaster genome, first published in 2000 through collaboration between Celera Genomics and the Drosophila Genome Projects, has provided a number of important contributions to genome research. By demonstrating the utility of methods such as whole-genome shotgun sequencing and genome annotation by a community "jamboree," the Drosophila genome established the precedents for the current paradigm used by most genome projects. Subsequent releases of the initial genome sequence have been improved by the Berkeley Drosophila Genome Project and annotated by FlyBase, the Drosophila community database, providing one of the highest-quality genome sequences and annotations for any organism. We discuss the impact of the growing number of genome sequences now available in the genus on current Drosophila research, and some of the biological questions that these resources will enable to be solved in the future.


High-Resolution, Non-Invasive Imaging of Upper Vocal Tract Articulators Compatible with Human Brain Recordings.
A complete neurobiological understanding of speech motor control requires determination of the relationship between simultaneously recorded neural activity and the kinematics of the lips, jaw, tongue, and larynx. Many speech articulators are internal to the vocal tract, and therefore simultaneously tracking the kinematics of all articulators is nontrivial--especially in the context of human electrophysiology recordings. Here, we describe a noninvasive, multi-modal imaging system to monitor vocal tract kinematics, demonstrate this system in six speakers during production of nine American English vowels, and provide new analysis of such data. Classification and regression analysis revealed considerable variability in the articulator-to-acoustic relationship across speakers. Non-negative matrix factorization extracted basis sets capturing vocal tract shapes allowing for higher vowel classification accuracy than traditional methods. Statistical speech synthesis generated speech from vocal tract measurements, and we demonstrate perceptual identification. We demonstrate the capacity to predict lip kinematics from ventral sensorimotor cortical activity. These results demonstrate a multi-modal system to non-invasively monitor articulator kinematics during speech production, describe novel analytic methods for relating kinematic data to speech acoustics, and provide the first decoding of speech kinematics from electrocorticography. These advances will be critical for understanding the cortical basis of speech production and the creation of vocal prosthetics.


Accessible chemical space for metal nitride perovskites.
Building on the extensive exploration of metal oxide and metal halide perovskites, metal nitride perovskites represent a largely unexplored class of materials. We report a multi-tier computational screening of this chemical space. From a pool of 3660 ABN3 compositions covering I-VIII, II-VII, III-VI and IV-V oxidation state combinations, 279 are predicted to be chemically feasible. The ground-state structures of the 25 most promising candidate compositions were explored through enumeration over octahedral tilt systems and global optimisation. We predict 12 dynamically and thermodynamically stable nitride perovskite materials, including YMoN3, YWN3, ZrTaN3, and LaMoN3. These feature significant electric polarisation and low predicted switching electric field, showing similarities with metal oxide perovskites and making them attractive for ferroelectric memory devices.


The Effects of Walking Workstations on Biomechanical Performance.
Prolonged sitting has been associated with negative health effects. Walking workstations have become increasingly popular in the workplace. There is a lack of research on the biomechanical effect of walking workstations. This study analyzed whether walking while working alters normal gait patterns. A total of 9 participants completed 4 walking trials at 2.4 and 4.0 km·h-1: baseline walking condition, walking while performing a math task, a reading task, and a typing task. Biomechanical data were collected using standard motion capture procedures. The first maximum vertical ground reaction force, stride width, stride length, minimum toe clearance, peak swing hip abduction and flexion angles, peak swing and stance ankle dorsiflexion, and knee flexion angles were analyzed. Differences between conditions were evaluated using analysis of variance tests with Bonferroni correction (P ≤ .05). Stride width decreased during the reading task at both speeds. Although other parameters exhibited significant differences when multitasking, these changes were within the normal range of gait variability. It appears that for short periods, walking workstations do not negatively impact gait in healthy young adults.


Muscadine grape skin extract inhibits prostate cancer cells by inducing cell-cycle arrest, and decreasing migration through heat shock protein 40.
Previously we demonstrated that muscadine grape skin extract (MSKE), a natural product, significantly inhibited androgen-responsive prostate cancer cell growth by inducing apoptosis through the targeting of survival pathways. However, the therapeutic effect of MSKE on more aggressive androgen-independent prostate cancer remains unknown. This study examined the effects of MSKE treatment in metastatic prostate cancer using complementary PC-3 cells and xenograft model. MSKE significantly inhibited PC-3 human prostate cancer cell tumor growth in vitro and in vivo. The growth-inhibitory effect of MSKE appeared to be through the induction of cell-cycle arrest. This induction was accompanied by a reduction in the protein expression of Hsp40 and cell-cycle regulation proteins, cyclin D1 and NF-kBp65. In addition, MSKE induced p21 expression independent of wild-type p53 induced protein expression. Moreover, we demonstrate that MSKE significantly inhibited cell migration in PC-3 prostate cancer cells. Overall, these results demonstrate that MSKE inhibits prostate tumor growth and migration, and induces cell-cycle arrest by targeting Hsp40 and proteins involved in cell-cycle regulation and proliferation. This suggests that MSKE may also be explored either as a neo-adjuvant or therapeutic for castration resistant prostate cancer.


[Pleiotropic effects of statins in systemic sclerosis].
To study anti-inflammatory and angioprotective properties of statins in patients with systemic sclerosis (SS). Fifty SS patients enrolled in the trial were divided into two groups: the study group included 30 patients who received standard therapy and atorvastatin in a dose 10-40 mg/day, the control group (n = 20) received standard therapy alone. The blood of the patients was studied with enzyme immunoassay for concentrations of highly sensitive C-reactive protein (hs-CRP), IL-6, VCAM-1, E-selectin, WF antigen, VEGF-A, VEGF-R2. The tests were made before the treatment and after 12 months of the treatment. The levels of hs-CRP, IL-6, VCAM-1, E-selectin, WF antigen, VEGF-A and VEGF-R2 were significantly higher in the study group than in healthy donors. The controls also had elevated levels of the above substances except IL-6 and VEGF-A which elevated but not significantly. A 12-month treatment in the study group resulted in a significant lowering of hs-CRP, IL-6, WF antigen, E-selectin, VEGF-A and VEGF-R2, the number of patients with elevated IL-6, WF antigen, VEGF-R2 also reduced while in the controls the parameters did not change. Atorvastatin has anti-inflammatory and angioprotective effects in SS patients.


Extracellular matrix molecules enhance the neurotrophic effect of Schwann cell-like differentiated adipose-derived stem cells and increase cell survival under stress conditions.
Since the first reports of induction of adipose-derived stem cells (ASC) into neuronal and glial cell phenotypes, expectations have increased regarding their use in tissue engineering applications for nerve repair. Cell adhesion to extracellular matrix (ECM) is a basic feature of survival, differentiation, and migration of Schwann cells (SC) during nerve regeneration, and fibronectin and laminin are two key molecules of this process. Interaction between ECM and SC-like differentiated ASC (dASC) could potentially improve the neurotrophic potential of the stem cells. We have investigated the effect of ECM molecules on SC-like dASC in terms of proliferation, adhesion, and cell viability. Fibronectin and laminin did not affect the proliferation of dASC when compared with cell adherent tissue culture plastic, but significantly improved viability and cell attachment when dASC were exposed to apoptotic conditions. To assess the influence of the ECM molecules on dASC neurotrophic activity, dASC were seeded onto ECM-coated culture inserts suspended above dorsal root ganglia (DRG) sensory neurons. Neurite outgrowth of DRG neurons was enhanced when dASC were seeded on fibronectin and laminin when compared with controls. When DRG neurons and dASC were in direct contact on the various surfaces there was significantly enhanced neurite outgrowth and coculture with laminin-conditioned dASC produced the longest neurites. Compared with primary SCs, dASC grown on laminin produced similar levels of neurite outgrowth in the culture insert experiments but neurite length was shorter in the direct contact groups. Anti β1 integrin blocking antibody could inhibit baseline and dASC evoked neurite elongation but had no effect on outgrowth mediated by laminin-conditioned dASC. ECM molecules had no effect on the levels of nerve growth factor and brain-derived neurotrophic factor secretion from dASC. The results of the study suggest that ECM molecules can significantly improve the potential of dASC for nerve regeneration.


Aortic valve stenosis in community medical practice: determinants of outcome and implications for aortic valve replacement.
To define the objective and subjective measures of aortic stenosis (AS) severity linked to survival after diagnosis in community practice. All 360 Olmsted County, Minnesota residents (74 ± 14 years; 44% men) with AS diagnosed from 1988 to 1997 by echocardiography and without life-threatening comorbid conditions were enrolled. The presentation at first diagnosis, outcomes (mortality, heart failure, cardiac surgery), and coherence of guideline-based criteria for severe AS were analyzed. The presentation was challenging. Cardiac symptoms were frequent (59%) and unassociated with the AS severity (all P > .13). Of the patients with severe AS, as determined by a valve area less than 1.0 cm(2), 67% had low gradient AS (≤40 mm Hg). An aortic valve area less than 1.0 cm(2) was the only objective measure independently determining survival (adjusted risk ratio, 1.81; 95% confidence interval [CI], 1.19-2.70; P < .01) and heart failure (adjusted risk ratio, 2.3; 95% CI, 1.3-4.0; P < .01), even in patients with low-gradient AS and/or an ejection fraction of 50% or greater. Excess mortality (vs expected mortality) occurred with an aortic valve area of less than 1.0 cm(2) (risk ratio, 1.78; 95% CI, 1.33-2.35; P < .001) even without symptoms (risk ratio, 1.65; 95% CI, 1.05-2.47; P = .02). Aortic valve replacement, ultimately performed in only 45% of those with an aortic valve area less than 1.0 cm(2), reduced mortality (risk ratio, 0.61; 95% CI, 0.39-0.94; P = .02) and heart failure (risk ratio, 0.29; 95% CI, 0.13-0.64; P < .01). In community practice, AS affects elderly patients, and its presentation is challenging owing to the high frequency of low-gradient severe AS despite a normal ejection fraction and because symptoms are frequently not specific to AS. Consequently, aortic valve replacement is seldom performed despite its considerable benefit. Physicians should be aware that an aortic valve area of less than 1.0 cm(2) predicts for unfavorable outcomes, irrespective of symptoms or gradient. Thus, such patients should undergo a thorough evaluation to detect those who could benefit from aortic valve replacement, despite their challenging presentation.


Abnormal eye movements in blepharospasm and involuntary levator palpebrae inhibition. Clinical and pathophysiological considerations.
We report on four patients with involuntary eyelid closure and eye movement disorders. Three were healthy until the onset of their illness and one had a mild generalized choreoathetosis and dystonia due to kernicterus. Electromyographic recording revealed solely blepharospasm in two patients and blepharospasm in combination with involuntary levator palpebrae inhibition in the other two. The eye movement abnormalities were clinically characterized by inability to fix gaze and short or prolonged episodes of uncontrollable eye deviations accompanied, in two patients, by diplopia in horizontal or vertical directions. These episodes occurred independently of a disorder of eyelid movement. Eye movement recordings with a double magnetic induction technique showed saccadic intrusions in horizontal directions. They consisted of highly frequent square wave jerks in three and sporadic macro-square wave jerks in two patients. There were also episodes of extraocular muscle dystonia, commonly known as oculogyric crises, resulting in involuntary upward eye deviation in all patients and lateral deviation in three patients. In one patient, nasal-ward deviations were sometimes restricted to one eye. We conclude that these abnormal eye movements do not necessarily point to a symptomatic form of dystonia and that they may limit the beneficial effect of botulinum toxin or surgical intervention in the therapeutic management of involuntary eyelid closure. We suggest that either basal ganglia, especially substantia nigra pars reticularis and the brainstem structures, especially the paramedian pontine reticular formation, or both, may be involved in the pathogenesis of these abnormal movements.


Effects of sterilized micropulverized barium sulfate suspension and meglumine iothalamate solution on the genitourinary tract of healthy male dogs after retrograde urethrocystography.
A sufficient quantity of 30% w/v sterile barium sulfate suspension was infused into the caudal portions of urinary tracts of 12 young adult healthy male Beagles to induce vesicoureteral reflux. A control group of dogs was infused with a 7.5% solution of meglumine iothalamate. The dogs were radiographed at 1, 6, 13, and 29 days later. Three dogs from each group were euthanatized and necropsied (1 each) after radiography at each of these times. In some dogs, radiopacity in the prostate, urinary bladder, and renal pelvic diverticula due to barium persisted throughout the 30-day observation period. Opacity at or beyond 24 hours was not found in dogs infused with iothalamate. Light microscopy revealed barium either free or within macrophages in the submucosa of the bladder, in glandular and stromal areas of the prostate, and in the kidney. Renal barium was limited to pelvic diverticula (3 dogs), in a medullary tubule (1 dog), and in cortical tubules (2 dogs). Significant lesions attributable to barium were not seen in kidneys of dogs in the barium group. Iothalamate induced focal to multifocal inflammatory responses in some prostates; barium was phagocytosed by macrophages. Focal transient inflammatory and ulcerative lesions induced by bladder distention were observed in the bladder and urethral mucosa in both groups.


Associations Between E-Cigarette Type, Frequency of Use, and Quitting Smoking: Findings From a Longitudinal Online Panel Survey in Great Britain.
E-cigarettes can be categorized into two basic types, (1) cigalikes, that are disposable or use pre-filled cartridges and (2) tanks, that can be refilled with liquids. The aims of this study were to examine: (1) predictors of using the two e-cigarette types, and (2) the association between type used, frequency of use (daily vs. non-daily vs. no use), and quitting. Online longitudinal survey of smokers in Great Britain was first conducted in November 2012. Of 4064 respondents meeting inclusion criteria at baseline, this study included (N = 1643) current smokers followed-up 1 year later. Type and frequency of e-cigarette use were measured at follow-up. At follow-up, 64% reported no e-cigarette use, 27% used cigalikes, and 9% used tanks. Among e-cigarette users at follow-up, respondents most likely to use tanks versus cigalikes included: 40-54 versus 18-24 year olds and those with low versus moderate/high education. Compared to no e-cigarette use at follow-up, non-daily cigalike users were less likely to have quit smoking since baseline (P = .0002), daily cigalike or non-daily tank users were no more or less likely to have quit (P = .3644 and P = .4216, respectively), and daily tank users were more likely to have quit (P = .0012). Whether e-cigarette use is associated with quitting depends on type and frequency of use. Compared with respondents not using e-cigarettes, daily tank users were more likely, and non-daily cigalike users were less likely, to have quit. Tanks were more likely to be used by older respondents and respondents with lower education.


A review of non-invasive imaging in extramammary Paget's disease.
Extramammary Paget's Disease (EMPD) is a rare intraepithelial adenocarcinoma that classically manifests with pruritic, erythematous and scaling plaques. The clinical picture frequently mimics inflammatory or infectious conditions and is thus commonly misdiagnosed. The assessment of tumour margins is equally challenging as tumours have a propensity to spread beyond clinically visible boundaries. Appropriate non-invasive diagnostic tools can assist in the early detection, diagnosis and management of EMPD. This paper will review the literature on non-invasive imaging modalities used in EMPD. Articles from the PubMed database were selected based on relevance to the topic of this review. Articles that were not specific to EMPD and non-invasive imaging were excluded. Search strategy is further described in the methods section below. Eighteen articles were selected for this review: six PET/CT, five reflectance confocal microscopy (RCM), two photodynamic diagnosis (PDD), two dermoscopy, two MRI and one optical coherence tomography (OCT) paper(s). Dermoscopy, PDD, RCM and OCT can help to distinguish malignant conditions, including EMPD, from benign conditions. RCM and OCT can identify atypical cells in real-time, and have the potential to improve the accuracy of surgical margins intraoperatively and overall management. Distinctive confocal characteristics of EMPD have been described using RCM. The sensitivity and specificity of these findings require additional validation. Radiographic techniques also play a central role in the diagnosis of EMPD and assessment of disease spread. PET/CT and MRI can detect primary disease, nodal and distant metastases, with superior delineation of disease spread on MRI. Limitations of PET/CT are mainly related to primary tumour thickness, and size and FDG-avidity of nodal and distant metastases. Limitations of MRI include the fact that few studies have examined its use in EMPD; additional research is warranted. Randomized controlled trials and large prospective studies evaluating the use of non-invasive imaging in EMPD are needed.


Ultrastructural diversity of the cellulase complexes of Clostridium papyrosolvens C7.
Transmission electron microscopy was used to investigate the ultrastructural features of diverse cellulase and cellulase-xylanase multiprotein complexes that are components of the cellulase-xylanase system of Clostridium papyrosolvens C7. The multiprotein complexes were separated by anion-exchange chromatography into seven biochemically distinguishable fractions (F1 to F7). Most individual F fractions contained, in relatively large numbers, an ultrastructurally recognizable type of particle that occurred only in smaller numbers, or not at all, in the other F fractions. It is suggested that these ultrastructurally distinct particles represent the biochemically distinct multiprotein complexes that constitute the cellulase-xylanase system of C. papyrosolvens C7. Some of the particles consisted of tightly packed globular components that appeared to be arranged in the shape of a ring with conical structures pointing out along its axis. Other particles had triangular, polyhedral, or star shapes. The major protein fraction (F4) almost exclusively contained particles consisting of loosely aggregated components, many of which appeared to be arranged along filamentous structures. The ultrastructural observations reported here support our previous conclusion that the cellulase-xylanase system of C. papyrosolvens C7 comprises at least seven different high-molecular-weight multiprotein complexes. Furthermore, results of this and earlier studies indicate that the interactions between C. papyrosolvens C7 and cellulose are different from those that have been described for Clostridium thermocellum.


A Qualitative Study on Knowledge and Attitude towards Risk Factors, Early Identification and Intervention of Infant Hearing Loss among Puerperal Mothers- A Short Survey.
Maternal active participation and their support are critical for the success of early hearing loss detection program. Erroneous maternal decisions may have large life long consequences on the infant's life. The mothers' knowledge and their attitudes towards infant hearing loss is the basis for their decisions. The present study was done to determine the mothers' knowledge and their attitude towards risk factors of infant hearing loss, its early identification and intervention and also awareness of effect of consanguinity on hearing loss. In this cross-sectional questionnaire survey study, a total of 100 mothers were interviewed using the questionnaire which consisted of three sections namely risk factors, early identification and early intervention of hearing loss. Chi-square test was used to establish relationship between consanguineous and non-consanguineous mother's responses to its effect on hearing loss. A p-value < 0.05 was considered as significant. Mothers' awareness was significantly high for visible causes (ear pain/discharge, head injury and slap to ear) of hearing loss. Positive attitude was seen for importance of screening programs and follow up testing. Moderate level of awareness was found on hazards of consanguinity and benefits of early identification. However, mothers were least aware of neonatal jaundice, NICU admission (>5 days), signs of late-onset and neural hearing loss, management of hearing loss, hearing aid fitting and therapy necessity, which might interfere in early detection and intervention of hearing loss. It is crucial to educate mothers on few risk factors and management of hearing loss to reduce its consequences.


Hemoglobin structure at higher levels of hemoglobin A1C in type 2 diabetes and associated complications.
Fourier transform infrared (FTIR) spectroscopy technique has not been used as a diagnostic tool for diabetes in clinical practice. This study was linked to structural changes in hemoglobin (Hb) in type 2 diabetes patients at higher levels of HbA1C using FTIR spectroscopy. Fifty-three diabetic patients from the Bahawal Victoria Hospital, Bahawalpur, Pakistan were categorized as group A (6% < HbA1C < 7%; n = 25) and group B (HbA1C ≥9%; n = 28). Another group (group N) of twenty blood samples was taken from healthy people from the Islamia University Bahawalpur, Pakistan. Data from all groups were collected from January 1, 2018 to March 31, 2019. The structure of Hb was studied by FTIR spectroscopy and impact of glucose on the fine structure of HbA1C was estimated. Hb secondary structure erythrocyte parameters were altered by changing glucose concentrations. From FTIR spectra of all three groups it was found that Hb structure was slightly altered in group A, but significantly changed in group B (P < 0.05). There was an increase in β-sheet structure and a reduction in α-helix structure at elevated levels of HbA1C (group B) in type 2 diabetes. We suggest that higher level of glycation reflected by increased HbA1C might be a contributing factor to structural changes in Hb in type 2 diabetic patients. FTIR spectroscopy can be a novel technique to find pathogenesis in type 2 diabetes.


Surface-protein interactions on different stainless steel grades: effects of protein adsorption, surface changes and metal release.
Implantation using stainless steels (SS) is an example where an understanding of protein-induced metal release from SS is important when assessing potential toxicological risks. Here, the protein-induced metal release was investigated for austenitic (AISI 304, 310, and 316L), ferritic (AISI 430), and duplex (AISI 2205) grades in a phosphate buffered saline (PBS, pH 7.4) solution containing either bovine serum albumin (BSA) or lysozyme (LSZ). The results show that both BSA and LSZ induce a significant enrichment of chromium in the surface oxide of all stainless steel grades. Both proteins induced an enhanced extent of released iron, chromium, nickel and manganese, very significant in the case of BSA (up to 40-fold increase), whereas both proteins reduced the corrosion resistance of SS, with the reverse situation for iron metal (reduced corrosion rates and reduced metal release in the presence of proteins). A full monolayer coverage is necessary to induce the effects observed.


The relationship between macroeconomic and industry-specific business cycle indicators and work-related injuries among Danish construction workers.
The current study examines and compares the relationship between both macroeconomic and industry-specific business cycle indicators, and work-related injuries among construction workers in Denmark using emergency department (ED) injury data and also officially reported injuries to the Danish Working Environment Authority (WEA). The correlations between ED and WEA injury data from the catchment area of Odense University Hospital during the period 1984-2010 were tested separately for variability and trend with two general macroeconomic indicators (gross domestic product and the Danish unemployment rate) and two construction industry-specific indicators (gross value added and the number of employees). The results show that injury rates increase during economic booms and decrease during recessions. However, the regression coefficients were generally weak for both the ED (range 0.14-0.20) and WEA injuries (range 0.13-0.36). Furthermore, although there is some variability in the strength of the relationship of the different business cycle indicators, the relationships are generally not stronger for the WEA injuries than for the ED injuries, except for general unemployment. Similarly, no substantial differences in strength of relation between industry-specific and macroeconomic indicators were identified. The study shows that there was no difference in the relationship between business cycle indicators, and WEA and ED injury data. This indicates that changes in reporting behaviour do not seem to play a major role in the relation between the business cycle and workplace injuries in a Danish context.


WDR72 Mutations Associated with Amelogenesis Imperfecta and Acidosis.
Dental enamel malformations, or amelogenesis imperfecta (AI), can be isolated or syndromic. To improve the prospects of making a successful diagnosis by genetic testing, it is important that the full range of genes and mutations that cause AI be determined. Defects in WDR72 (WD repeat-containing protein 72; OMIM *613214) cause AI, type IIA3 (OMIM #613211), which follows an autosomal recessive pattern of inheritance. The defective enamel is normal in thickness, severely hypomineralized, orange-brown stained, and susceptible to attrition. We identified 6 families with biallelic WDR72 mutations by whole exome sequence analyses that perfectly segregated with the enamel phenotype. The novel mutations included 3 stop-gains [NM_182758.2: c.377G>A/p.(Trp126*), c.1801C>T/p.(Arg601*), c.2350A>T/p.(Arg784*)], a missense mutation [c.1265G>T/p.(Gly422Val)], and a 62,138-base pair deletion (NG_017034.2: g.35441_97578del62138) that removed WDR72 coding exons 3 through 13. A previously reported WDR72 frameshift was also observed [c.1467_1468delAT/p.(Val491Aspfs*8)]. Three of the affected patients showed decreased serum pH, consistent with a diagnosis of renal tubular acidosis. Percentiles of stature and body weight varied among 8 affected individuals but did not show a consistent trend. These studies support that WDR72 mutations cause a syndromic form of AI and improve our ability to diagnose AI caused by WDR72 defects.


My paper 10 years later: infective endocarditis in the intensive care unit.
Although the recent literature contains plenty of studies concerning all aspects of infective endocarditis (IE), very few focus on severe IE requiring admission to the ICU. In 2004, we published a report on the clinical spectrum and prognostic factors in 228 consecutive critically ill patients with IE. Septic shock, neurological complications and immunocompromised state were independently associated with in-hospital mortality. Cardiac surgery during the acute phase of EI was associated with better survival. A lot of information has been accumulated during the past 10 years on management of IE. Although three sets of blood cultures allow the identification of about 90% of cases, culture-negative IE still remains a diagnostic challenge. Blood-polymerase chain reaction in valve tissue may yield a microbiologic diagnosis. New imaging techniques such as positron emission tomography computed tomography (PET-CT) have shown additive value in patients with an intracardiac device or valvular prosthesis. Systematic cerebral magnetic resonance imaging can lead to modification of therapeutic plans. The decision to operate and the timing of cardiac surgery should take into account the presence of congestive heart failure, neurological complications, renal failure and multiorgan dysfunction syndrome. In 2011 and 2013, we published the results of a multicentre prospective observational study of 198 ICU patients with left-sided IE and confirmed that cardiac surgery was associated with better outcome. The strongest independent predictor of post-operative mortality was the pre-operative multiorgan failure score. Neurological failure also represented a major determinant of mortality, regardless of the mechanism of neurological complication. In the present paper, we propose algorithms to optimize the medico-surgical approach.


In vivo induction of T-cell hyporesponsiveness and alteration of immunological cells of bone marrow grafts using granulocyte colony-stimulating factor.
Granulocyte colony-stimulating factor (G-CSF)-primed bone marrow (G-BM) transplantation is associated with a low incidence of graft-versus-host disease (GVHD) and has been used successfully in patients with human leukocyte antigen (HLA) matched/mismatched donors. This study evaluated the function of T cells and the quantities of immunological cells of G-BM. Bone marrow was obtained from fifteen donors by aspiration. Lymphocyte proliferation ability, interferon-gamma (IFN-gamma) and interleukin-4 (IL-4) secreted by T cells were determined using a monotetrazolium (MTT) assay and sandwich enzyme-linked immunosorbent assay (ELISA), respectively. T-cell subgroups, dendritic cell (DC) subsets, CD4(+)CD25(+) regulatory T cells and the expression of CD28/CD80/CD86 molecules on monocytes, B and T cells were analyzed using flow cytometry. G-CSF treatment decreased the quantities of IFN-gamma secretion dramatically (p=0.007) and IL-4 moderately (p=0.027), leading to higher ratios of IL-4/IFN-gamma (p=0.004). We confirmed T-cell hyporesponsiveness and lower expression of CD28/CD80/CD86 on monocytes, B and T cells. The absolute values of lymphocytes, T cell subgroups, CD3+CD4-CD8- cells, CD8+CD28- cells and B cells in bone marrow grafts were similar before and after G-CSF treatment. The number of monocytes per microliter was increased 2.13-fold, while the numbers of CD4(+)CD25(+) regulatory T cells were unchanged. DC2 were preferentially increased. Our results suggest that bone marrow T-cell hyporesponsiveness could be induced and that the increase of monocytes and DC2 and the downregulation of CD28/CD80/CD86 co-stimulatory signals were produced by in vivo use of G-CSF; this may be related to the preferential increase of monocytes and DC2 and the downregulation of CD28/CD80/CD86 co-stimulatory signals.


Digital bioethics: introducing new methods for the study of bioethical issues.
The online space has become a digital public square, where individuals interact and share ideas on the most trivial to the most serious of matters, including discussions of controversial ethical issues in science, technology and medicine. In the last decade, new disciplines like computational social science and social data science have created methods to collect and analyse such data that have considerably expanded the scope of social science research. Empirical bioethics can benefit from the integration of such digital methods to investigate novel digital phenomena and trace how bioethical issues take shape online.Here, using concrete examples, we demonstrate how novel methods based on digital approaches in the social sciences can be used effectively in the domain of bioethics. We show that a digital turn in bioethics research aligns with the established aims of empirical bioethics, integrating with normative analysis and expanding the scope of the discipline, thus offering ways to reinforce the capacity of bioethics to tackle the increasing complexity of present-day ethical issues in science and technology. We propose to call this domain of research in bioethics digital bioethics.


Targeted disruption of retinoic acid receptor alpha (RAR alpha) and RAR gamma results in receptor-specific alterations in retinoic acid-mediated differentiation and retinoic acid metabolism.
F9 embryonic teratocarcinoma stem cells differentiate into an epithelial cell type called extraembryonic endoderm when treated with retinoic acid (RA), a derivative of retinol (vitamin A). This differentiation is presumably mediated through the actions of retinoid receptors, the RARs and RXRs. To delineate the functions of each of the different retinoid receptors in this model system, we have generated F9 cell lines in which both copies of either the RAR alpha gene or the RAR gamma gene are disrupted by homologous recombination. The absence of RAR alpha is associated with a reduction in the RA-induced expression of both the CRABP-II and Hoxb-1 (formerly 2.9) genes. The absence of RAR gamma is associated with a loss of the RA-inducible expression of the Hoxa-1 (formerly Hox-1.6), Hoxa-3 (formerly Hox-1.5), laminin B1, collagen IV (alpha 1), GATA-4, and BMP-2 genes. Furthermore, the loss of RAR gamma is associated with a reduction in the metabolism of all-trans-RA to more polar derivatives, while the loss of RAR alpha is associated with an increase in metabolism of RA relative to wild-type F9 cells. Thus, each of these RARs exhibits some specificity with respect to the regulation of differentiation-specific gene expression. These results provide an explanation for the expression of multiple RAR types within one cell type and suggest that each RAR has specific functions.


Therapeutic Role of Harmalol Targeting Nucleic Acids: Biophysical Perspective and in vitro Cytotoxicity.
Harmalol, a beta carboline alkaloid, shows remarkable importance in the contemporary biomedical research and drug discovery programs. With time, there is emerging interest in search for better anti-cancer drugs of plant origin with high activity and lower toxicity. Most of the chemotherapeutic agents due to their non-specific target and toxicity on active healthy cells, use is often restricted, necessitating search for newer drugs having greater potentiality. The review highlighted the interaction of harmalol with nucleic acids of different motifs as sole target biomolecules and in vitro cytotoxicity of the alkaloid in human cancer cell lines with special emphasis on its apoptotic induction ability. Binding study and in vitro cytotoxicity was performed using several biophysical techniques and biochemical assays, respectively. Data from competition dialysis, UV and fluorescence spectroscopic analysis, circular dichroism, viscometry and isothermal calorimetry shows binding and interaction of harmalol with several natural and synthetic nucleic acids, both DNA and RNA, of different motifs. Furthermore, apoptotic hallmarks like internucleosomal DNA fragmentation, membrane blebbing, cell shrinkage, chromatin condensation, change of mitochondrial membrane potential, comet tail formation and ROS (reactive oxygen species) dependent cytotoxicity being analyzed in the harmalol treated cancer cells. These results stating the therapeutic role of harmalol, will lead to the interesting knowledge on the cytotoxicity, mode, mechanism, specificity of binding and correlation between structural aspects and energetics enabling a complete set of guidelines for design of new drugs.


[Development of long-term post-tetanic potentiation and changes in S-100 protein contents in hippocampal slices of rats in different functional states].
Male rats of the strains with low (LE) high excitability (HE) of the nervous system have been used in this study. Half of the animals of each strain were neurotized in accordance with the Hecht's scheme. In the hippocampal slices of the non-neurotized LE rats there was a significant increase of the populational spike amplitude during development of LTP as compared with the opposite group of the animals. The LTP formation in the LE strain of rats caused a decrease in the S-100 protein content in the water-soluble, and an increase in the membrane-bound fraction of the protein. Similar results we have observed with the non-inbred Wistar rats but not with the HE strain of the animals. The levels of the water-soluble S-100 protein fraction were also higher in the hippocampuses and entorenal cortices, but not in the cerebellae of the LE strain, as compared with the HE strain of the rats. No differences have been found in the membrane-bound fraction of S-100 protein.


Altered stratum corneum barrier and enhanced percutaneous immune responses in filaggrin-null mice.
Loss-of-function mutations in filaggrin are major predisposing factors for atopic dermatitis. Although various reports suggest a critical role for filaggrin in stratum corneum (SC) barrier formation, the lack of filaggrin-null (Flg(-/-)) mice has hampered detailed in vivo analysis of filaggrin's functions. We sought to generate Flg(-/-) mice and to assess the effect of filaggrin loss on SC barrier function and percutaneous immune responses. We generated Flg(-/-) mice using gene targeting and assessed the morphology, hydration, mechanical strength, and antigen permeability of their SC. Percutaneous immune responses were evaluated through irritant- and hapten-induced contact hypersensitivity studies and by measuring humoral responses to epicutaneous sensitization with protein antigen. Newborn Flg(-/-) mice exhibited dry scaly skin. Despite marked decreases in natural moisturizing factor levels, which are filaggrin degradation products, SC hydration and transepidermal water loss were normal. Microscopic analyses suggested premature shedding of SC layers, and indeed, increased desquamation under mechanical stress was demonstrated. Loss of keratin patterns, which are critical for corneocyte stabilization, is likely attributable to fragility in the Flg(-/-) SC. Antigens penetrated the Flg(-/-) SC more efficiently, leading to enhanced responses in hapten-induced contact hypersensitivity and higher serum levels of anti-ovalbumin IgG(1) and IgE. Complete filaggrin deficiency led to altered barrier integrity and enhanced sensitization, which are important factors in early-phase atopic dermatitis. Flg(-/-) mice should provide a valuable tool to further explore additional factors the dysfunction of which leads to uncontrolled inflammation in patients with atopic diseases.