Infratentorial choroid plexus tumors in children.
Choroid plexus tumors (CPTs) are rare pediatric intracranial neoplasms, and mostly occur in the lateral ventricle. CPTs located in the infratentorial location are considered to be rare in the pediatric population. We present a series of eight patients treated in the last decade at our institution focusing on clinical presentations and their outcome after excision. We performed an institutional retrospective review of patients who underwent surgical resection of infratentorial CPTs during the period from 2008 to 2017. Patients' charts were reviewed for demographic data, clinical presentation, surgical treatment, and follow-up. There were eight patients (6 females and 2 males), with mean age for the cohort at presentation was 9.0 years. They represent 75% of 12 CPTs of all locations treated at the same period in our institution. These 8 infratentorial CPTs were in the fourth ventricle in seven, and in the cerebellopontine angle (CPA) in one. Seven patients had choroid plexus papillomas (WHO grade I) and 1 had an atypical choroid plexus papilloma (WHO grade II). Gross total resection was attempted in all patients. However, two of 3 patients with fourth ventricle floor invasion had subtotal resection with a thin layer of tumor left on the floor. The remaining 6 had a gross total resection. Six patients with preoperative hydrocephalus had a perioperative external ventricular drainage but none required permanent shunting after tumor resection. None showed recurrence/tumor progression without adjuvant therapy during the follow-up period of 20 months to 11 years. Infratentorial dominance among pediatric CPTs in this series contradicts previous reports. Infratentorial CPTs are amenable to surgical resection. Unresected small residuals due to invasion to the fourth ventricle floor showed no regrowth during 2 to 3 years follow-up without adjuvant therapy. However, these patients with incomplete resection need watchful observations.


Forebrain connectivity of the prefrontal cortex in the marmoset monkey (Callithrix jacchus): an anterograde and retrograde tract-tracing study.
The cortical and subcortical forebrain connections of the marmoset prefrontal cortex (PFC) were examined by injecting the retrograde tracer, choleratoxin, and the anterograde tracer, biotin dextran amine, into four sites within the PFC. Two of the sites, the lateral and orbital regions, had previously been shown to provide functionally dissociable contributions to distinct forms of behavioral flexibility, attentional set-shifting and discrimination reversal learning, respectively. The dysgranular and agranular regions lying on the orbital and medial surfaces of the frontal lobes were most closely connected with limbic structures including cingulate cortex, amygdala, parahippocampal cortex, subiculum, hippocampus, hypothalamus, medial caudate nucleus, and nucleus accumbens as well as the magnocellular division of the mediodorsal nucleus of the thalamus and midline thalamic nuclei, consistent with findings in the rhesus monkey. In contrast, the granular region on the dorsal surface closely resembled area 8Ad in macaques and had connections restricted to posterior parietal cortex primarily associated with visuospatial functions. However, it also had connections with limbic cortex, including retrosplenial and caudal cingulate cortex as well as auditory processing regions in the superior temporal cortex. The granular region on the lateral convexity had the most extensive connections. Based on its architectonics and functionality, it resembled areas 12/45 in macaques. It had connections with high-order visual processing regions in the inferotemporal cortex and posterior parietal cortex, higher-order auditory and polymodal processing regions in the superior temporal cortex. In addition it had extensive connections with limbic regions including the amygdala, parahippocampal cortex, cingulate, and retrosplenial cortex.


Visuomotor sensitivity to visual information about surface orientation.
We measured human visuomotor sensitivity to visual information about three-dimensional surface orientation by analyzing movements made to place an object on a slanted surface. We applied linear discriminant analysis to the kinematics of subjects' movements to surfaces with differing slants (angle away form the fronto-parallel) to derive visuomotor d's for discriminating surfaces differing in slant by 5 degrees. Subjects' visuomotor sensitivity to information about surface orientation was very high, with discrimination "thresholds" ranging from 2 to 3 degrees. In a first experiment, we found that subjects performed only slightly better using binocular cues alone than monocular texture cues and that they showed only weak evidence for combining the cues when both were available, suggesting that monocular cues can be just as effective in guiding motor behavior in depth as binocular cues. In a second experiment, we measured subjects' perceptual discrimination and visuomotor thresholds in equivalent stimulus conditions to decompose visuomotor sensitivity into perceptual and motor components. Subjects' visuomotor thresholds were found to be slightly greater than their perceptual thresholds for a range of memory delays, from 1 to 3 s. The data were consistent with a model in which perceptual noise increases with increasing delay between stimulus presentation and movement initiation, but motor noise remains constant. This result suggests that visuomotor and perceptual systems rely on the same visual estimates of surface slant for memory delays ranging from 1 to 3 s.


Smoking, Hypertension, and GG Genotype of the IL-6 rs1800796 Polymorphism are Independent Risk Factors for Abdominal Aortic Aneurysm in Han Population.
Recent researches have investigated the link between the rs1800796 polymorphism (-572G/C) in the interleukin-6 (IL-6) gene and the risk of abdominal aortic aneurysm (AAA). However, no Chinese studies have addressed the association between the risk of AAA and this polymorphism. This case-control study included 153 AAA patients and 205 controls. Diabetes mellitus, hypertension, dyslipidemia, and coronary artery disease (CAD) status were collected. The AAA and control groups were 69.20 ± 7.56 and 68.50 ± 7.12 years old, respectively. The analysis revealed that the G allele and GG genotype of the IL-6 rs1800796 polymorphism were remarkedly correlated with an elevated risk of AAA. The GG genotype displayed an association with the risk of AAA after adjustment for gender, body mass index, age, drinking, and smoking. Subgroup analyses indicated that this polymorphism elevated the risk of AAA among males, individuals aged ≥70 years, smokers, drinkers, individuals with a body mass index (BMI) ≥25 kg/m2, patients without diabetes, and patients with hypertension. Univariate and multivariate analyses indicated that smoking, hypertension, and the GG genotype (rs1800796 polymorphism) were independently related with the risk of AAA. The IL-6 rs1800796 polymorphism increases the risk of AAA. In addition, smoking, hypertension, and the GG genotype of the rs1800796 polymorphism are independent risk factors for AAA. Further studies of Chinese populations are needed to validate these findings.


Existence of functional alpha1A- and alpha1D- but no alpha1B-adrenoceptor subtypes in rat common carotid arteries.
Using the cannula inserting method, vasoconstrictor responses to alpha1-adrenoceptor agonists (noradrenaline [NA], phenylephrine [PE] and methoxamine [ME]) and effects of alpha1-adrenoceptor antagonists (WB4101, chloroethylclonidine [CEC] and BMY7378) were investigated in isolated and perfused rat common carotid arteries. The rank order of agonist potency and efficacy was NA = PE > ME. Either WB4101 or BYM7378 inhibited NA- and PE-induced constrictions in a dose-related manner. CEC did not inhibit the NA- and PE-induced responses. The ME-induced responses were also significantly blocked by either WB4101 or BMY7378. From these results, it is concluded that there are functional alpha1A- and alpha1D-adrenoceptor subtypes in rat common carotid arteries, but no functional alpha1B subtype.


Primary omental-mesenteric myxoid hamartoma of the mesoappendix incidentally detected after abdominal trauma in a child: report of a case.
A 14-year-old boy was brought to our hospital with abdominal pain and nausea after suffering a blow to the abdomen. A mass was felt in the right hypogastrium, and the patient was hospitalized for possible hematoma resulting from the abdominal trauma. Initially, we treated him conservatively and observed his course, but on the 20th day after trauma, enhanced computed tomography showed that the area of strong enhancement in the tumor was unchanged. Superior mesenteric angiography showed findings indicative of a pseudoaneurysm caused by the trauma, and surgery was performed 26 days after the injury. Laparotomy revealed a tumor with a clear boundary, thought to originate in the mesoappendix, without any sign of pseudoaneurysm. Histopathological examination confirmed that the tumor was an omental-mesenteric myxoid hamartoma. The patient had an uneventful postoperative course and was discharged from hospital on the 12th day after surgery. More than 5 years have elapsed since the operation and no sign of recurrence or metastasis has been recognized.


Role of human hepatocyte nuclear factor 4alpha in the expression of drug-metabolizing enzymes and transporters in human hepatocytes assessed by use of small interfering RNA.
Hepatocyte nuclear factor 4alpha (HNF4alpha) is an important transcription factor in hepatic gene expression. Here, we have investigated the role of HNF4alpha in the expression of drug-metabolizing enzymes and transporters in human hepatocytes using an adenovirus expressing human HNF4alpha-small interfering RNA (hHNF4alpha-siRNA). The hHNF4alpha-siRNA effectively reduced the mRNA and nuclear protein levels of hHNF4alpha in a concentration-dependent manner. The hHNF4alpha-siRNA also decreased the mRNA levels of CYP2A6, CYP2B6, CYP2C8, CYP2C9, CYP2C19, CYP2D6, CYP3A4, UGT1A1, UGT1A9, SULT2A1, ABCB1, ABCB11, ABCC2, OATP1B1 and OCT1, as well as those of PXR and CAR. To discern the role of these nuclear receptors, we co-infected hepatocytes with hHNF4alpha-siRNA and PXR- or CAR-expressing adenovirus. The hHNF4alpha-siRNA-induced reductions of the enzyme and transporter mRNA levels were not restored except CYP2B6 mRNA levels, which were returned to the control level by overexpressing CAR. Furthermore, although hHNF4alpha-siRNA did not significantly affect the fold-induction of CYP2B6, CYP2C8, CYP2C9, or CYP3A4 mRNA levels following treatment with CYP inducers, the levels in hHNF4alpha-suppressed cells fell significantly compared to the control. These results suggest that HNF4alpha plays a dominant role in the expression of drug-metabolizing enzymes and transporters in human hepatocytes, and that HNF4alpha expression levels is a possible determinant for inter-individual variations in the expression of these enzymes and transporters.


Binding of rat brain hexokinase to recombinant yeast mitochondria: identification of necessary physico-chemical determinants.
The association of rat brain hexokinase with heterologous recombinant yeast mitochondria harboring human porin (Yh) is comparable to that with rat liver mitochondria in terms of cation requirements, cooperativity in binding, and the effect of amphipathic compounds. Mg2+, which is required for hexokinase binding to all mitochondria, can be replaced by other cations. The efficiency of hexokinases, however, depends on the valence of hydrophilic cations, or the partition of hydrophobic cations in the membrane, implying that these act by reducing a prohibitive negative surface charge density on the outer membrane rather than fulfilling a specific structural requirement. Macromolecular crowding (using dextran) has dual effects. Dextran added in excess increases hexokinase binding to yeast mitochondria, according to the porin molecule they harbor. This effect, significant with wild-type yeast mitochondria, is only marginal with Yh as well as rat mitochondria. On the other hand, an increase in the number of hexokinase binding sites on mitochondria is also observed. This increase, moderate in wild-type organelles, is more pronounced with Yh. Finally, dextran, which has no effect on the modulation of hexokinase binding by cations, abolishes the inhibitory effect of amphipathic compounds. Thus, while hexokinase binding to mitochondria is predetermined by the porin molecule, the organization of the latter in the membrane plays a critical role as well, indicative that porin must associate with other mitochondrial components to form competent binding sites on the outer membrane.


Sámi traditional medicine and complementary and alternative medicine - a descriptive study of use within the Sámi population of Sweden.
Traditional medicine has been practised for millennia in the Sámi population, based on a Sámi worldview and cosmology, which includes natural remedies, prayers, drums and yoik singing. During the Christianisation of the Sámi during the seventeenth and eighteenth centuries, these practices were condemned. In recent years, however, a revival of Sámi culture has occurred and so has the practice of Sámi traditional medicine (STM) and the use of complementary alternative medicine (CAM). The aim of this study is to map the prevalence and use of STM and CAM among Sámi in Sweden today. The study population consisted of 3641 Sámi from the whole of Sweden, who had participated in the population-based cross-sectional survey Sámi Health on Equal Terms (SámiHET) in 2021. Our results show that women are more prone to use both STM and CAM than men and that younger persons are more likely to use STM and CAM than elderly persons. STM is more often used in the northern parts of Sápmi compared to the southern parts as well as a lower use of CAM in the north. This might be due to the stronger Sámi identity and easier access to traditional Sámi healers/helpers in the north as well as limited access to CAM services.


Rehydration with fluid of varying tonicities: effects on fluid regulatory hormones and exercise performance in the heat.
This study examined the effects of rehydration (Rehy) with fluids of varying tonicities and routes of administration after exercise-induced hypohydration on exercise performance, fluid regulatory hormone responses, and cardiovascular and thermoregulatory strain during subsequent exercise in the heat. On four occasions, eight men performed an exercise-dehydration protocol of approximately 185 min (33 degrees C) to establish a 4% reduction in body weight. Following dehydration, 2% of the fluid lost was replaced during the first 45 min of a 100-min rest period by one of three random Rehy treatments (0.9% saline intravenous; 0.45% saline intravenous; 0.45% saline oral) or no Rehy (no fluid) treatment. Subjects then stood for 20 min at 36 degrees C and then walked at 50% maximal oxygen consumption for 90 min. Subsequent to dehydration, plasma Na(+), osmolality, aldosterone, and arginine vasopressin concentrations were elevated (P < 0.05) in each trial, accompanied by a -4% hemoconcentration. Following Rehy, there were no differences (P > 0.05) in fluid volume restored, post-rehydration (Post-Rehy) body weight, or urine volume. Percent change in plasma volume was 5% above pre-Rehy values, and plasma Na(+), osmolality, and fluid regulatory hormones were lower compared with no fluid. During exercise, skin and core temperatures, heart rate, and exercise time were not different (P > 0.05) among the Rehy treatments. Plasma osmolality, Na(+), percent change in plasma volume, and fluid regulatory hormones responded similarly among all Rehy treatments. Neither a fluid of greater tonicity nor the route of administration resulted in a more rapid or greater fluid retention, nor did it enhance heat tolerance or diminish physiological strain during subsequent exercise in the heat.


Population variation in sexual selection and its effect on size allometry in two dung fly species with contrasting sexual size dimorphism.
Body size is one of the most important quantitative traits under evolutionary scrutiny. Sexual size dimorphism (SSD) in a given species is expected to result if opposing selection forces equilibrate differently in both sexes. We document variation in the intensity of sexual and fecundity selection, male and female body size, and thus SSD among 31 and 27 populations of the two dung fly species, Scathophaga stercoraria and Sepsis cynipsea, across Switzerland. Whereas in S. cynipsea females are larger, the SSD is reversed in S. stercoraria. We comprehensively evaluated Fairbairn and Preziosi's (1994) general, three-tiered scenario, hypothesizing that sexual selection for large male size is the major driving force of SSD allometry within these two species. Sexual selection intensity on male size in the yellow dung fly, S. stercoraria, was overall positive, greater, and more variable among populations than fecundity selection on females. Also, sexual selection intensity in a given population correlated positively with mean male body size of that population for both the field-caught fathers and their laboratory-reared sons, indicating a response to selection. In S. cvnipsea, sexual selection intensity on males was lower overall and significantly positive, about equal in magnitude, but more variable than fecundity selection on females. However, there was no correlation between the intensity of sexual selection and mean male body size among populations. In both species, the laboratory-reared offspring indicate genetic differentiation among populations in body size. Despite fulfillment of all key prerequisites, at least in S. stercoraria, we did not find hypoallometry for SSD (Rensch's rule, i.e., greater evolutionary divergence in male size than female size) for the field-caught parents or the laboratory-reared offspring: Female size was isometric to male size in both species. We conclude that S. cynipsea does not fit some major requirements of Fairbairn and Preziosi's (1994) scenario, whereas for S. stercoraria we found partial support for it. Failure to support Rensch's rule within the latter species may be due to phylogenetic or other constraints, power limitations, erroneous estimates of sexual selection, insufficient genetic isolation of populations, or sex differences in viability selection against large size.


Linezolid Pharmacia Corp.
Linezolid is an oxazolidinone developed by Pharmacia (formerly Pharmacia & Upjohn) for the treatment of multi-resistant Gram-positive infections [187765,317456]. It binds to ribosomal 50S subunits, most likely within domain V within the 23S rRNA peptidyl transferase and a secondary interaction with the 30S subunit. This results in inhibition of the initiation of protein translation at an early point, which is probably N-formylmethionyl-tRNA [335843]. No direct action on DNA or RNA synthesis has been observed [220169]. Linezolid resistance due to a 23S rRNA mutation may emerge in Enterococci during therapy with this antimicrobial, and may be associated with clinical failure [368652]. Following FDA approval, linezolid was launched in May 2000 [368526,368652]. In April 2000, the FDA approved linezolid injection, tablets and oral suspension for the treatment of patients with infections caused by Gram-positive bacteria. It is indicated for adults in the treatment of nosocomial pneumonia, community-acquired pneumonia (CAP), complicated and uncomplicated skin and skin structure infections and vancomycin-resistant enterococcus (VRE) infections caused by methicillin-resistant Staphylococcus aureus (MRSA), VRE faecium and penicillin-susceptible Streptococcus pneumoniae [363503]. The FDA, however, did not grant Pharmacia indications for linezolid in the treatment of CAP due to either penicillin-resistant S aureus (PRSA) or MRSA. In May 2000, Merrill Lynch predicted sales for 2000 to be US $50 million, rising to US $760 million in 2004 [366910]. In February 2000, P&U predicted that peak sales of the drug had the potential to reach in excess of US $750 million [358429]. In February 1999, Morgan Stanley Dean Witter predicted sales of US $40 million in 2000 rising to US $275 million in 2005 [319855]. In December 1998, Deutsche Bank predicted sales of US $100 million in 2000 rising to US $300 million in 2002 [316769].


Transgene insertion on mouse chromosome 6 impairs function of the uterine cervix and causes failure of parturition.
The molecular mechanisms controlling the initiation of parturition remain largely undefined. We report a new animal model in which parturition does not occur. A line of mice expressing a human apolipoprotein B (APOB) gene fail to deliver their young if the transgene is present in homozygous (Tg/Tg), but not in heterozygous (Tg/Wt), form. Cloning and mapping of the transgene insertion locus indicate that 10 copies of the 80-kilobase APOB genomic fragment inserted into mouse chromosome 6 result in a small, 390-base pair deletion of mouse genomic DNA. Nine other lines expressing the transgene have normal labor, suggesting that transgene insertion in this mutant line disrupted a mouse gene crucial for successful parturition. The pathophysiology of parturition failure in these animals was defined using physiological, endocrinological, and morphological techniques. Results indicate that luteolysis occurs in Tg/Tg mice but is delayed by 1 day. Delivery did not occur in mutant mice at term after spontaneous luteolysis or even after removal of progesterone action by ovariectomy or antiprogestin treatment. Biomechanical and functional studies of the uterus and cervix revealed that the primary cause of failed parturition in Tg/Tg mice was not inadequate uterine contractions of labor but, rather, a rigid, inelastic cervix at term that was abnormally rich in neutrophils and tissue monocytes. Characterization of the transgene insertional mutant, Tg/Tg, indicates that progesterone withdrawal is insufficient to complete parturition in the presence of inadequate cervical ripening at term.


[Estimated number of orphans due to maternal AIDS in Brazil, 1987-99].
This study proposes a procedure to estimate the number of orphans due to maternal AIDS. The procedure estimates the number of orphans by calendar year, multiplying the cumulative fertility rate by the number of AIDS deaths among women aged 15-49 years. Because the procedure refers to the number of children that are alive, the estimate is adjusted by contemplating the proportion of pediatric AIDS cases due to vertical transmission and the probability of survival in the 5-9-year age group. To estimate the number of AIDS orphans in Brazil from 1987 to 1999, the procedure was applied by stratifying according to geographical region, taking into account the differences in regional fertility rates, completeness of death reporting, and misclassification of AIDS-related deaths as due to other causes. The total number of cumulative AIDS orphans for 1987-99 was estimated at approximately 30,000.


Managing multimorbidity: Profiles of integrated care approaches targeting people with multiple chronic conditions in Europe.
In response to the growing populations of people with multiple chronic diseases, new models of care are currently being developed in European countries to better meet the needs of these people. This paper aims to describe the occurrence and characteristics of various types of integrated care practices in European countries that target people with multimorbidity. Data were analysed from multimorbidity care practices participating in the Innovating care for people with multiple chronic conditions (ICARE4EU) project, covering all 28EU Member States, Iceland, Norway and Switzerland. A total of 112 practices in 24 countries were included: 65 focus on patients with any combination of chronic diseases, 30 on patients with a specific chronic disease with all kinds of comorbidities and 17 on patients with a combination of specific chronic diseases. Practices that focus on a specific index disease or a combination of specific diseases are less extensive regarding the type, breadth and degree of integration than practices that focus on any combination of diseases. The latter type is more often seen in countries where more disciplines, e.g. community nurses, physiotherapists, social workers, work in the same primary care practice as the general practitioners. Non-disease specific practices put more emphasis on patient involvement and provide more comprehensive care, which are important preconditions for person-centered multimorbidity care.


Prevalence and risk factors of obesity in school children in Antalya, Turkey.
To determine the prevalence and risk factors of obesity and to obtain the age and gender-specific body mass index (BMI) percentiles in a cohort of children aged 6-17 years, living in the province of Antalya, Turkey. The study included 15 schools throughout the city center of Antalya, Turkey during the period November 2002-March 2003. A total of 2465 school children (boys 1233, girls 1232) aged 6-17 years were chosen using a population based stratified cluster sampling method. We calculated the BMI (kg/m2) by measuring the weight and standing height. Overweight was defined as BMI between 85th and 95th percentile, and obesity as BMI above the 95th percentile. A questionnaire was distributed to the parents to determine obesity-related risk factors. The overall prevalence of obesity was 3.6% while overweight was 14.3%. According to gender, the prevalence of obesity in boys was 3.9% and overweight was 12.8%, while in girls, obesity was 3.2% and overweight was 15.8%. We found that obesity might be related with some factors such as number of regular meals, number of siblings, high birth weight, having computer at home, skipping breakfast and high socioeconomic status of parents. There is no difference in obesity prevalence among school children according to gender, but the mean BMI of girls is significantly higher than that of boys. Obesity prevalence among children in Antalya is very low compared to Europe and the United States.


[Risk factors for urothelial carcinoma: drinking measures, smoking and other life style-related risk factors--results of the Berlin Urothelial Study (BUS)].
With the exception of smoking and several occupational exposures there is little knowledge about risk factors for urothelial cancer. A case control study in the area of former West Berlin was performed from 1990-1995 to investigate the role of several lifestyle risk factors, such as smoking, drinking behaviour and regular intake of analgesics and laxatives. The study includes 647 hospital-based incident cases with bladder cancer (n = 571), renal pelvis cancer (n = 51), and ureter cancer (n = 25), and 647 population-based controls which were matched individually by sex and age. Data analyses were carried out using standard methods for case control studies (conditional multiple logistic regression analysis). Odds ratios (OR) and 95% confidence intervals (CI) were applied as effect parameter. Statistically significantly increased odds ratios were observed for current smoking (OR: 3.46, 95% CI: 2.50-4.78), previous but now abandoned smoking (OR: 1.51, 95% CI: 1.09-2.81), and for regular intake of laxatives (OR: 2.52, 95% CI: 1.56-4.09). Furthermore, an increased risk for urothelial cancer was observed for daily consumption of three and more litres of cold drinks (OR: 2.65 95% CI: 1.12-6.24). The results underline that lifestyle factors other than smoking may contribute to a higher risk of urothelial cancer.


Usutu Virus Antibodies in Blood Donors and Healthy Forestry Workers in the Lombardy Region, Northern Italy.
Usutu virus (USUV), a member of the genus Flavivirus, is known to circulate at low prevalence in Northern Italy, and has been reported to cause overt infection. USUV was first reported in Europe in 2001, but a retrospective study showed that it has been present in Italy at least since 1996. Seroprevalence data for USUV antibodies in sera are being collected in different European countries, showing circulation at low prevalence in human populations. Interestingly, two consecutive studies in Northern Italy indicate a possible increase in the presence of the virus, from 0% to 0.23% seroprevalence in blood donors. In this study, antibodies against USUV were measured in 3 consecutive blood samples collected from October 2014 to December 2015 from 33 forestry workers in the Po river valley, while samples from 200 blood donors from the same geographical area were tested in parallel. Neutralizing and IgG antibodies were found in six forestry workers (18.1%) and in two blood donors (1%). Our results indicate that USUV circulation in the examined area, part of a highly populated region in Northern Italy, is higher than expected. Healthy subjects exhibit a higher prevalence than what was found in a previous report in an adjoining region (0.23%), while the population at risk shows a much higher prevalence value (18.1%).


Specificity of human lymphocyte complement receptors.
Erythrocytes, bone marrow-derived lymphocytes, monocytes, and granulocytes were shown to have a receptor activity for C4. Theis C4 receptor activity was studied in relation to the previously identified C3b and C3d receptors. By assay for inhibition of rosette formation by fluid-phase complement (C), only two different lymphocyte C receptors were demonstrated. The immune adherence receptor, the only one of the two shared in common with erythrocytes, was specific for C4 or the C3c region of C3b, but was unreactive with C3d. The other lymphocyte receptor, the C3d receptor, was specific for C3d fragments, but would also react to a lesser extent with the C3d region of uncleaved C3b. ThC3d receptor did not react with either C3c or C4. This specificity of the C3d receptor allowed certain cells which contained only C3d receptors to form rosettes with EAC1-3b and EAC1-3d, but not with EAC14. However, because C3d receptors bound EAC1-3d or C3d fragments more firmly than they did EAC1-3b or C3b fragments, many other types of cells containing only C3d receptors, formed rosettes with EAC1-3d but not with EAC1-3b. Erythrocytes and those lymphocytes which contained only immune adherence receptors, formed rosettes with EAC14 and EAC1-3D but not with EAC1-3d. A double-label assay was devised for the simultaneous detection of both types of C receptors on individual lymphocytes. This assay involved fluorescence labeling of one of the two C receptors with soluble C fragments in combination with the usual rosette method for labeling the other type of C receptor. With this double-label assay, it was observed that the two different lymphocyte C receptors capped independently and thus were located on different molecules which could each move through the fluid membrane matrix independently of the other.


L-Dopa in small peptides: an amazing functionality to form supramolecular materials.
l-Dopa (3,4-dihydroxyphenylalanine) is a chiral amino acid generated via biosynthesis from l-tyrosine in plants and some animals. The presence of multiple interacting sites makes l-Dopa a multifunctional building block for the preparation of supramolecular materials. The possibility to form hydrogen bonds and the presence of the aromatic ring allow l-Dopa molecules to interact through a series of non-covalent interactions. The additional presence of the catechol moiety really makes this compound unique: not only does it have implications in the self-assembly process of Dopa itself and with other substrates, but also it highly increases the number of applications of the final material, since it works as an antioxidant, radical trapper, metal chelator, reducing agent and adhesive. l-Dopa and catechol containing derivatives have been extensively introduced inside both synthetic and natural polymers to obtain amazing functional materials. In this review we report the preparation of small peptides containing l-Dopa, focusing on the supramolecular materials that can be obtained with them, ranging from fibrils to fibres, gels, films and coatings, all having the different applications mentioned above and many others.


Photothermal conversion of gold nanoparticles for uniform pulsed laser warming of vitrified biomaterials.
Pulsed laser (ms, 1064 nm) gold nanoparticle (GNP) heating has been used recently to achieve fast (>10 000 000 °C min-1) warming of vitrified droplets using gold nanorods (GNRs) as photon-absorbers. To maximize the viability of biomaterials in vitrified droplets, the droplets must be warmed as uniformly as possible. A potential approach to such warming is to use an appropriate combination of photon-absorption and -scattering to distribute heat more uniformly throughout a droplet. To investigate this, 2 plasmonic gold nanorods (GNRs), 1 hollow gold nanoshell, and 2 silica-core gold nanoshells (GNSs) were synthesized and characterized under 1064 nm laser irradiation in water, propylene glycol, and protein-rich (egg white) solutions. Using a modified cuvette laser calorimetry experiment with complementary Monte Carlo modeling, the GNSs were found to have higher per-particle absorption and scattering cross sections, while the GNRs had higher photothermal conversion efficiency, absorption efficiency, and Au mass normalized absorption cross sections. In the characterization, the GNSs with larger scattering-to-absorption ratios could have ∼30% over-estimation of photothermal conversion efficiency if scattering and reabsorption inside the solution were not considered, while GNRs with lower ratios were less impacted. Combined Monte Carlo and COMSOL simulations were used to predict the specific absorption rate (W m-3) and heating behavior of GNP-loaded hemispherical droplets, thereby demonstrating that the GNS case with higher scattering-to-absorption ratio achieved more uniform heating than the GNR case. Interestingly, further tuning of the scattering and absorption coefficients of the hemispherical GNP-loaded droplet within the model suggests the ability to obtain an optimal scattering-to-absorption ratio for uniform heating. These results show the importance of considering the reabsorption of scattered light to accurately characterize the photothermal conversion efficiency of GNP solutions during laser irradiation. We also show that the relative scattering and absorption properties of the nanoparticles can be designed to promote both rapid and uniform laser rewarming of vitrified droplets for application in cryopreservation.


Associations between weather-related data and influenza reports: A pilot study and related policy implications.
The purpose of this retrospective, correlational pilot study was to explore the relationship between historical weekly weather data including temperature, dew point, humidity, barometric pressure, visibility, and cloud cover compared to weekly influenza-like illness reports over a four year period. Climate and weather-related conditions may affect the viral activity and transmission of influenza, although this relationship has not been widely studied in nursing. Some research suggests that there are causal links between cold temperatures, low indoor humidity, minimal sun exposure, and influenza outbreaks. Additionally, rapid weather variability in a warming climate can increase influenza epidemic risk. Data from a local public health district were extracted and used to correlate with weekly weather averages for the area. Findings showed that current influenza reports are significantly associated with temperature and visibility, both lagged two weeks. Though more research is needed, nurses must understand, recognize, and act upon weather and climate factors that affect the health of populations. With a greater understanding of the relationship between weather and influenza-like illness, nurses and other healthcare providers can potentially work to respond to and mitigate the consequences of weather-related illness as well as anticipate and prepare for increased flu burden. Furthermore, nurses can remain engaged in climate protective initiatives and policy development at their local community and/or organizational levels to underscore and advocate for the needs of populations and groups they serve.


Incidence of Guillain-Barré Syndrome After COVID-19 Vaccination in the Vaccine Safety Datalink.
Postauthorization monitoring of vaccines in a large population may detect rare adverse events not identified in clinical trials such as Guillain-Barré syndrome (GBS), which has a background rate of 1 to 2 per 100 000 person-years. To describe cases and incidence of GBS following COVID-19 vaccination and assess the risk of GBS after vaccination for Ad.26.COV2.S (Janssen) and mRNA vaccines. This cohort study used surveillance data from the Vaccine Safety Datalink at 8 participating integrated health care systems in the United States. There were 10 158 003 participants aged at least 12 years. Data analysis was performed from November 2021 to February 2022. Ad.26.COV2.S, BNT162b2 (Pfizer-BioNTech), or mRNA-1273 (Moderna) COVID-19 vaccine, including mRNA vaccine doses 1 and 2, December 13, 2020, to November 13, 2021. GBS with symptom onset in the 1 to 84 days after vaccination, confirmed by medical record review and adjudication. Descriptive characteristics of confirmed cases, GBS incidence rates during postvaccination risk intervals after each type of vaccine compared with the background rate, rate ratios (RRs) comparing GBS incidence in the 1 to 21 vs 22 to 42 days postvaccination, and RRs directly comparing risk of GBS after Ad.26.COV2.S vs mRNA vaccination, using Poisson regression adjusted for age, sex, race and ethnicity, site, and calendar day. From December 13, 2020, through November 13, 2021, 15 120 073 doses of COVID-19 vaccines were administered to 7 894 989 individuals (mean [SE] age, 46.5 [0.02] years; 8 138 318 doses received [53.8%] by female individuals; 3 671 199 doses received [24.3%] by Hispanic or Latino individuals, 2 215 064 doses received [14.7%] by Asian individuals, 6 266 424 doses received [41.4%] by White individuals), including 483 053 Ad.26.COV2.S doses, 8 806 595 BNT162b2 doses, and 5 830 425 mRNA-1273 doses. Eleven cases of GBS after Ad.26.COV2.S were confirmed. The unadjusted incidence rate of GBS per 100 000 person-years in the 1 to 21 days after Ad.26.COV2.S was 32.4 (95% CI, 14.8-61.5), significantly higher than the background rate, and the adjusted RR in the 1 to 21 vs 22 to 42 days following Ad.26.COV2.S was 6.03 (95% CI, 0.79-147.79). Thirty-six cases of GBS after mRNA vaccines were confirmed. The unadjusted incidence rate per 100 000 person-years in the 1 to 21 days after mRNA vaccines was 1.3 (95% CI, 0.7-2.4) and the adjusted RR in the 1 to 21 vs 22 to 42 days following mRNA vaccines was 0.56 (95% CI, 0.21-1.48). In a head-to-head comparison of Ad.26.COV2.S vs mRNA vaccines, the adjusted RR was 20.56 (95% CI, 6.94-64.66). In this cohort study of COVID-19 vaccines, the incidence of GBS was elevated after receiving the Ad.26.COV2.S vaccine. Surveillance is ongoing.


Fluid collections associated with renal allografts.
In a series of 50 consecutive renal allograft patients, 4 developed lymphoceles and 1 developed a seroma which later became infected. In no instance could the fluid collection be held responsible for deterioration in graft function. Two of the lymphoceles occurred with ureteric strictures and perirenal abscess was associated with acute rejection. The significance of a perirenal collection with deteriorating renal function is discussed.


Lissencephaly and the molecular basis of neuronal migration.
Migration of post-mitotic neurons from the ventricular zone to the cortical plate during embryogenesis comprises one of the most critical stages in brain development. Deficiency of this process often results in major brain malformations, including human lissencephaly (smooth brain). Since discovery of the first genetic cause of lissencephaly, deletions of chromosome 17p13.3 in Miller-Dieker syndrome, rapid progress in our understanding of neuronal migration has been made based on advances in both brain imaging technology and molecular genetics. This progress has resulted in a new system of classification that began with pathological descriptions and has evolved to include patterns on brain imaging, causative genes and most recently the molecular pathways and proposed modes of migration involved. In this review, we summarize current knowledge regarding five genes that cause or contribute to human lissencephaly, including LIS1, 14-3-3 epsilon, DCX, RELN and ARX. Each of these is associated with a characteristic pattern of malformation that involves the cerebral cortex and sometimes other brain structures. Based on detailed genotype-phenotype analysis, we can now infer the most likely causative gene based on brain imaging and other clinical findings, and inversely are becoming able to predict clinical severity based on the specific mutations detected. We also hypothesize, for the first time, a relationship between the specific type of lissencephaly observed and deficiency of specific modes of neuronal migration.


Do diabetic foot infections with methicillin-resistant Staphylococcus aureus differ from those with other pathogens?
There is controversy as to whether or not diabetic foot infections (DFIs) caused by methicillin-resistant Staphylococcus aureus (MRSA) are associated with worse outcomes than DFIs caused by other pathogens. To address this issue we performed a nonsystematic literature search of published articles in English language journals seeking studies reporting on the outcomes of DFIs related to their microbiology. We retrieved 48 articles published from 1999 to 2013 that described a total of 7771 cases of DFI. The overall proportion of DFIs with an isolate of S aureus was about 30%; just over one third of these (11% of all cases) were MRSA strains. Among the DFI cases caused by MRSA 1543 were episodes of soft tissue infections and 113 of osteomyelitis, while non-MRSA organisms caused 5761 soft tissue infections and 354 cases of osteomyelitis. Only 5 of the included articles attempted a comparison between DFI caused by MRSA and those caused by other pathogens, with no clear differences noted. The median total duration of antibiotic therapy for DFI caused by MRSA was 26 days, of which a median of 10 days was given intravenously. Only a few articles reported the proportion of patients with a recurrence, but they often did not differentiate between MRSA and non-MRSA cases. Four publications reported a worse functional or microbiological outcome in MRSA, compared to non-MRSA, cases, but the findings were variable and differences did not seem to be significant. Many trials failed to adjust for case-mix or to definitively demonstrate a relationship between microbiology and outcomes. Few of the articles specifically commented on whether the MRSA isolates were health care- or community-acquired strains. Notwithstanding the substantial limitations of the available literature, there does not appear to be a need for any special treatment for DFI caused by MRSA. The current guidelines for treating according to established international recommendations seem appropriate.


[A technique of constructing human chromosomal band-specific probe pools using degenerate oligonucleotide primer].
To establish a rapid and efficient technique of constructing human chromosomal band specific probe pools and their libraries. A modified method of combining chromosome microdissection with degenerate oligonucleotide primed PCR(DOP-PCR) was used. 3p23-p26, 3q21-q22 and 4p12- p16 band from human chromosomes were microdissected and amplified as probe pools. The origins of the PCR products were determined by chromosome fluorescence in situ hybridization. The PCR products and pUC19 were digested by Xho I and Sal I respectively, and linke up. The DH5alpha were transformed by the recombinated vectors as the specific band libraries. The inserts were digested by EcoR I and Hind III, then measured by electrophoretic analysis. And the copies of inserts were identified by in situ bacterial colony hybridization with genomic DNA. All the three probe pools showed the special yellow-green signals in their microdissection responsible bands. The sizes of DOP-PCR products ranged from 300bp to 1800bp. 3q21-q22 probe pool generated about 1.2 x 10(4) clones. The average size of inserts was about 420bp by analysis of 30 positive clones. The rate of single-copy and low-repeated sequences was about 81%(178/220), while the rate of middle-repeated and high- repeated sequences was about 19%(42/220). The results proved that the modified microdissection combining DOP-PCR technique provided a simple and efficient method to construct the human chromosome band-specific probe pools and might contribute to gene cloning and complete sequencing of human genome.


Cilostamide produces hyperpolarization associated with K(ATP) channel activation, but does not augment endothelium-derived hyperpolarization in rat mesenteric arteries.
1. Non-nitric oxide/prostaglandin-mediated endothelium-derived hyperpolarization (EDH) is considered to be mediated, in part, by gap junctions and it has been suggested that cAMP increases endothelium-derived hyperpolarizing factor (EDHF)-mediated relaxation through the modulation of gap junctions. Cilostamide, which inhibits phosphodiesterase III, has been suggested to augment EDHF-type relaxation by increasing the concentration of cAMP. 2. In the present study, we investigated the effect of cilostamide on EDH per se in mesenteric arteries of Wistar rats using a conventional microelectrode technique. 3. The resting membrane potential of the mesenteric arteries was significantly more negative in the presence of 10(-6) mol/L cilostamide compared with control conditions. Furthermore, EDH in response to 10(-6) mol/L acetylcholine (ACh) in the presence of 10(-5) mol/L indomethacin and 10(-4) mol/L N(G)-nitro-L-arginine was decreased in the presence of 10(-6) mol/L cilostamide by approximately 5 and 3.5 mV in proximal and distal arteries, respectively. 4. Glibenclamide (10(-5) mol/L), an ATP-sensitive potassium channel (K(ATP)) inhibitor, abolished the hyperpolarization to 10(-6) mol/L cilostamide. Furthermore, in the presence of glibenclamide, ACh-induced EDH was unaffected by cilostamide, suggesting that the inhibition of ACh-induced hyperpolarization by cilostamide in the absence of glibenclamide may be due to the smaller driving force for hyperpolarization because of the more negative membrane potential under such conditions. 5. The findings of the present study suggest that cilostamide produces hyperpolarization by activating K(ATP) channels, presumably by increasing cAMP. However, cilostamide alone may not directly affect EDH.


[Production of Candida lipolytica protoplasts].
Optimal conditions were found for the production and isolation of the protoplasts of Candida lipolytica. The maximum amount of the protoplasts was produced after 90 minutes of the incubation with a crude preparation of the enzyme from Helix pomatia (100 mg/g wet biomass). Longer incubation results in lysis of the protoplasts and structural damages of the intracellular components. The yield of the protoplasts does not depend on the nature of stabilizing agent. A decrease in the stabilizer concentration increases the yield of the protoplasts four times. Preliminary treatment of the yeast cells with a 0.1 M solution of SH compounds (cysteine, beta-mercaptoethanol) does not increase the yield of the protoplasts; and 0.2 M solution of these compounds decreased the yield of the protoplasts.


Alpha 1 antitrypsin deficiency: a study of the relationship between the Pi system and genetic markers.
Twenty-four members (4 generations) of a family with alpha 1 antitrypsin deficiency were studied in an attempt to determine the chromosomal location of the Pi system locus. Three alpha 1 antitrypsin alleles (PiM, PiI, and PiZ) and five phenotypes (MM, MZ, MI, IZ, and ZZ) were detected in family members. The quinacrine fluorescent banding technique was successfully utilized to reveal eight polymorphic chromosomal markers in family members. Eight red cell antigens and HL-A antigens were identified for each family member. No linkage between the Pi system and chromosomal markers, four polymorphic red cell antigens, and HL-A antigens was detected. On the basis of this family study, the Pi locus as defined by alpha 1 antitrypsin deficiency does not appear to be on chromosomes 2, 3, 13, 14, 21, or 22 within measurable distance of the markers used.


Sampling saddle points on a free energy surface.
Many problems in biology, chemistry, and materials science require knowledge of saddle points on free energy surfaces. These saddle points act as transition states and are the bottlenecks for transitions of the system between different metastable states. For simple systems in which the free energy depends on a few variables, the free energy surface can be precomputed, and saddle points can then be found using existing techniques. For complex systems, where the free energy depends on many degrees of freedom, this is not feasible. In this paper, we develop an algorithm for finding the saddle points on a high-dimensional free energy surface "on-the-fly" without requiring a priori knowledge the free energy function itself. This is done by using the general strategy of the heterogeneous multi-scale method by applying a macro-scale solver, here the gentlest ascent dynamics algorithm, with the needed force and Hessian values computed on-the-fly using a micro-scale model such as molecular dynamics. The algorithm is capable of dealing with problems involving many coarse-grained variables. The utility of the algorithm is illustrated by studying the saddle points associated with (a) the isomerization transition of the alanine dipeptide using two coarse-grained variables, specifically the Ramachandran dihedral angles, and (b) the beta-hairpin structure of the alanine decamer using 20 coarse-grained variables, specifically the full set of Ramachandran angle pairs associated with each residue. For the alanine decamer, we obtain a detailed network showing the connectivity of the minima obtained and the saddle-point structures that connect them, which provides a way to visualize the gross features of the high-dimensional surface.


Comparative hemodynamic responses to chlorpromazine, nitroprusside, nitroglycerin, and trimethaphan immediately after open-heart operations.
The hemodynamic effects of intravenous chlorpormazine, nitroprusside, nitroglycerin, and trimethaphan camsylate were studied in 51 patients with high mean arterial pressures immediately after open-heart operations. Chlorpromazine was given by bolus intravenous injection to 24 patients (average dose 10.3 mg); nitroprusside (17 patients), nitroglycerin (8 patients), and trimethaphan (12 patients) were administered by constant intravenous infusion at average doses of 77 mug/min, 59 mug/min, and .097 mg/min, respectively. Measured or derived variables included right atrial pressure, pulmonary artery pressure, left atrial pressure, systemic arterial pressure, heart rate (HR), cardiac index (CI), stroke index (SI), stroke work index (SWI), and systemic vascular resistance index (SVRI). All four vasodilators significantly reduced systemic and intracardiac pressures and SWI (P less than 0.01). Associated changes in left ventricular pumping performance, however, differed importantly between groups. Chlorpromazine caused a significant rise (+19%) in HR with preservation of SI; thus, CI rose significantly (P less than 0.01). Only nitroprusside, however, resulted in enhancement of SI (P less than 0.05) at the lowered left atrial pressure; CI increased by 19% (P less than 0.01), HR rose minimally (6.5%), and calculated SVRI diminished 33% (P less than 0.01). Both nitroglycerin and trimethaphan caused decreases in SI and CI. These results indicate that in general, among the vasodilators studied, nitroprusside is associated with the most favorable hemodynamic responses in early postoperative cardiac surgical patients.


Characterization of reutericyclin produced by Lactobacillus reuteri LTH2584.
Lactobacillus reuteri LTH2584 exhibits antimicrobial activity that can be attributed neither to bacteriocins nor to the production of reuterin or organic acids. We have purified the active compound, named reutericyclin, to homogeneity and characterized its antimicrobial activity. Reutericyclin exhibited a broad inhibitory spectrum including Lactobacillus spp., Bacillus subtilis, B. cereus, Enterococcus faecalis, Staphylococcus aureus, and Listeria innocua. It did not affect the growth of gram-negative bacteria; however, the growth of lipopolysaccharide mutant strains of Escherichia coli was inhibited. Reutericyclin exhibited a bactericidal mode of action against Lactobacillus sanfranciscensis, Staphylococcus aureus, and B. subtilis and triggered the lysis of cells of L. sanfranciscensis in a dose-dependent manner. Germination of spores of B. subtilis was inhibited, but the spores remained unaffected under conditions that do not permit germination. The fatty acid supply of the growth media had a strong effect on reutericyclin production and its distribution between producer cells and the culture supernatant. Reutericyclin was purified from cell extracts and culture supernatant of L. reuteri LTH2584 cultures grown in mMRS by solvent extraction, gel filtration, RP-C(8) chromatography, and anion-exchange chromatography, followed by rechromatography by reversed-phase high-pressure liquid chromatography. Reutericyclin was characterized as a negatively charged, highly hydrophobic molecule with a molecular mass of 349 Da. Structural characterization (A. Höltzel, M. G. Gänzle, G. J. Nicholson, W. P. Hammes, and G. Jung, Angew. Chem. Int. Ed. 39:2766-2768, 2000) revealed that reutericyclin is a novel tetramic acid derivative. The inhibitory activity of culture supernatant of L. reuteri LTH2584 corresponded to that of purified as well as synthetic reutericyclin.


"I think maybe 10 years seems a bit long." Beliefs and attitudes of women who had never used intrauterine contraception.
To explore, in a general practice setting, the concerns, beliefs and attitudes about intrauterine contraception (IUC) reported by women, who had never used the methods. We used a sequential mixed-method (QUAL/quant) approach. A pragmatic, self-selecting sample of 30 women, aged 18-46 years, who had never used IUC), was recruited through seven general practices in South East England. Themes arising from qualitative interviews were used to construct a quantitative survey, completed by a pragmatic sample of 1195 women, aged 18-49 years, attending 32 general practices in the same region, between February and August 2015. Qualitative themes were concerns about the long-acting nature of IUC, concerns about body boundaries, and informal knowledge of IUC, especially 'friend of a friend' stories. Women were not sure if the devices can be removed before their full 5- or 10-year duration of use, and felt that these timeframes did not fit with their reproductive intentions. Quantitative survey data showed that the most commonly endorsed concerns among never-users were painful fitting (55.8%), unpleasant removal of the device (60.1%), and concern about having a device 'inside me' (60.2%). To facilitate fully informed contraceptive choice, information provided to women considering IUC should be tailored to more fully address the concerns expressed by never-users, particularly around the details of insertion and removal, and concerns about the adverse, long-term effects of the device. Women need to be reassured that IUC can be removed and fertility restored at any time following insertion. Trial registration NIHR CRN portfolio; 15912.


The potential clinical relevance of anatomical structures and variations of the maxillary sinus for planned sinus floor elevation procedures: A retrospective cone beam computed tomography study.
Complications arising from sinus floor elevation (SFE) with lateral approach surgery can be avoided by means of maxillary sinus examination in the preoperative period. To investigate anatomical variations in the maxillary sinus by making use of cone beam computed tomography (CBCT) in terms of SFE with a lateral approach. Two hundred twenty eight maxillary sinuses in 114 patients were included. Parameters such as the position and diameter of the posterior superior alveolar artery (PSAA) canal, the frequency and localization of the septa or accessory maxillary ostium (AMO), and lateral wall thickness values were subjected to statistical analysis. Septa were observed in 35.1% of sinuses. The majority of septa were determined in the middle region (48.8%). Additionally, 71.1% of PSAA canals were intraosseous, generally exceeding 1 mm in diameter (68.9%). The shortest mean perpendicular distance between the sinus floor and the PSAA canal was determined in the first molar region (9.22 ± 5.66 mm). Similarly, the highest mean sinus lateral wall thickness was determined in the first molar region, in the area 3 mm distant from the sinus floor (2.42 mm ± 0.88 mm). AMO was detected in 40.8% of sinuses. CBCT-guided treatment planning may be beneficial prior to SFE procedures in order to avoid surgical complications.


Enzyme-Treated Zizania latifolia Ethanol Extract Protects from UVA Irradiation-Induced Wrinkle Formation via Inhibition of Lysosome Exocytosis and Reactive Oxygen Species Generation.
Ultraviolet A (UVA) is a risk factor for photoaging and wrinkle formation. Zizania latifolia is an herbaceous perennial plant. It contains many bioactive compounds such as tricin that show antioxidative and anti-inflammatory effects. The aim of this study was to investigate the antiwrinkle effect of a mixture of hydrolytic enzyme (cellulase, hemicellulase and pectinase)-treated Z. latifolia extract (ZLE) and tricin on UVA-irradiated human dermal fibroblasts (HDFs) and SKH-1 hairless mice. Treatment of UVA-irradiated HDF cells with ZLE and tricin significantly decreased UVA induced-plasma membrane rupture, generation of ROS, expression levels of total and secreted lysosomal associated membrane protein (LAMP-1), cathepsin B and metalloproteinases (MMPs) and inhibited NF-κB activation. In the animal study, UVA-damaged epidermal and dermal tissues were repaired by the ZLE and tricin treatments. Administration of ZLE or tricin to UVA-irradiated animals recovered skin surface moisture and collagen fiber in dermal tissue. Treatment of ZLE or tricin decreased wrinkle formation, secretion of MMPs and expression levels of vascular endothelial growth factor (VEGF) and cathepsin B, and increased the expression level of collagen-1 in UVA-irradiated animals. Overall, the ZLE and tricin treatments decreased the skin damage induced by UVA irradiation via inhibition of lysosomal exocytosis and ROS generation. Therefore, ZLE and tricin are promising as antiwrinkle and antiphotoaging agents.


Mechanisms of the secretory and motor responses of the Brunner's gland region of the intestines to duodenal acidification.
The secretory and motor responses of the Brunner's gland region of the duodenum to luminal acidification were examined in rabbits anesthetized with urethan. Isomotic solutions of sodium chloride (pH 7.2) were perfused continuously through adjacent in situ segments. The pH of the perfusate of the proximal segment was changed to 2.0 for 30 min. Perfusion pressure (motor response) and volume and hexosamine concentration (secretory response) of the effluent were recorded for 3 h. The motor and secretory responses to luminal acidification were examined after intravenous (atropine) or intraluminal (lidocaine) pretreatment. Responses to intravenous infusions of serotonin or secretin were determined, also. Both atropine and lidocaine eliminated the initial motor and secretory responses of the proximal segment. Lidocaine eliminated the delayed secretory response of the distal segment. Serotonin caused initial motor and secretory responses, but secretin caused a delayed secretory response only. These results suggest that duodenal acidification elicits a two-phase increase in Brunner's gland secretion, the first being motor-dependent and the second motor-independent. The initial motor response was mediated by a local reflex composed of cholinergic and perhaps tryptaminergic receptors. The delayed secretory response was mediated by local and nonlocal, possibly hormonal, factors. Increased duodenal motility may provide a vehicle for the rapid expulsion of mucus, and thereby serve an important role in the function of the Brunner's glands.


Geraniin suppresses tumor cell growth and triggers apoptosis in human glioma via inhibition of STAT3 signaling.
Natural phytochemicals are attracting increasing interest as anticancer agents. The aim of this study is to evaluate the therapeutic potential of geraniin, a major ellagitannin extracted from Geranium sibiricum L., in human glioma. Human U87 and LN229 glioma cells were treated with different concentrations of geraniin, and cell viability, apoptosis, and gene expression were assessed. The involvement of STAT3 signaling in the action of geraniin was examined. We found that geraniin treatment for 48 h significantly (P < 0.05) impaired the phosphorylation of STAT3 and reduced the expression of downstream target genes Bcl-xL, Mcl-1, Bcl-2, and cyclin D1. Exposure to geraniin led to a concentration-dependent decline in cell viability and increase in apoptosis in glioma cells, but had no significant impact on the viability of normal human astrocytes. Measurement of caspase-3 activity showed that geraniin-treated U87 and LN229 cells showed a 1.8-2.5-fold higher caspase-3 activity than control cells. Overexpression of constitutively active STAT3 significantly (P < 0.05) reversed geraniin-mediated growth suppression and apoptosis, which was accompanied by restoration of Bcl-xL, Mcl-1, Bcl-2, and cyclin D1 expression. In an xenograft tumor mouse model, geraniin treatment significantly retarded tumor growth and induced apoptosis. Western blot analysis confirmed the suppression of STAT3 phosphorylation in glioma xenograft tumors by geraniin. Taken together, these data suggest that geraniin exerts growth-suppressive and pro-apoptotic effects on glioma cells via inhibition of STAT3 signaling and may have therapeutic benefits in malignant gliomas.


Polypeptide-binding membrane receptors: analysis and classification.
Polypeptide receptors on mammalian plasma membranes can be categorized on the basis of function. The binding of ligand by class I receptors results in changes in cell metabolism or behavior. Hormone-receptor interactions typify this group. The binding of ligand by class II receptors in ligand internalization. Although changes in cellular activity may result from metabolism of the internalized ligand, the interaction between ligand and class II receptor does not itself lead to alterations in cell behavior. Class II receptors include those for low-density lipoproteins and for alpha-macroglobulin-protease complexes. Although receptors within each category are chemically disparate, they show striking similarities in behavior. Analysis of the behavioral patterns of receptors in each category reveals insights into receptor physiology and allows for a prospective analysis of receptor characteristics.


The antioxidant, anti-inflammatory and analgesic activity effect of ethyl acetate extract from the flowers of Syringa pubescens Turcz.
Syringa Pubescens Turcz. (SP), a member of the Oleaceae family, is a species of plant known as Syringa. Flowers, as the medicinal part, are commonly used in the treatment of hepatitis and tonsillitis. The research was the first to assess the antioxidant and anti-inflammatory potential of different parts of SP flowers (SPF) in vitro. The most promising fraction was ethyl acetate fraction of SP flower (SPFEA). The antioxidant, anti-inflammatory and analgesic activities of SPFEA were further studied, and the chemical components were identified. HPLC was used to identify the major components in various fraction of SPF. DPPH and ABTS + radical scavenging assays as well as FRAP test and β-carotene bleaching test were employed to assess the antioxidant potential of SPF fraction in vitro. The inhibitory effect on NO production in LPS-treated RAW264.7 cells and heat-induced protein denaturation test were used to evaluate the anti-inflammatory potential of SPF fraction. Further analysis of the biological activity of SPFEA was performed. Acute toxicity test was conducted to assess the toxicity of SPFEA. The anti-inflammatory effect was assessed by utilizing xylene induced ear edema model, carrageenan-induced foot edema model and peritonitis model in vivo. The analgesic effect of SPFEA was evaluated using hot plate test, tail immersion test, formaldehyde test as well as acetic acid-induced abdominal writhing pain experiment in vivo. In carrageenan induced foot edema model, ELISA kits were employed to measure levels of inflammation factors (NO, TNF-α, IL-6, COX-2, IL-1β) in foot tissue as well as MDA, CAT, SOD, GSH-PX levels in liver tissue. HPLC results showed that there were significant differences in bioactive substances among different fractions of SPF, and SPFEA was rich in bioacitve components. Compared with other fractions of SPF, SPFEA exhibited better antioxidant and anti-inflammatory abilities. The 3000 mg/kg SPFEA group in mice had no obvious side effects. The xylene-induced ear edema model, carrageenan-induced foot edema and peritonitis models demonstrated that the SPFEA had significant anti-inflammatory effect. Moreover, inflammation factors including NO, TNF-α, IL-6, COX-2, IL-1β were significantly reduced in SPFEA groups in foot tissue induced by carrageenan. Additionally, SPFEA effectively decreased liver tissue oxidative stress levels (MDA, SOD, GSH-PX and CAT). The bioactivities of SPFEA demonstrated a clear dose-dependent relationship. The results of the hot plate test, tail immersion test, formaldehyde test and acetic acid-induced abdominal writhing pain experiments indicated the SPFEA possessed an excellent analgesic effect, and this effect was in dose-dependent manner. The study provides a scientific foundation for understanding the pharmacological action of SPFEA. It has been indicated that SPFEA has excellent antioxidant, analgesic and anti-inflammatory effects.


Epigenomic Regulation of Schwann Cell Reprogramming in Peripheral Nerve Injury.
The rapid and dynamic transcriptional changes of Schwann cells in response to injury are critical to peripheral nerve repair, yet the epigenomic reprograming that leads to the induction of injury-activated genes has not been characterized. Polycomb Repressive Complex 2 (PRC2) catalyzes the trimethylation of lysine 27 of histone H3 (H3K27me3), which produces a transcriptionally repressive chromatin environment. We find that many promoters and/or gene bodies of injury-activated genes of mature rat nerves are occupied with H3K27me3. In contrast, the majority of distal enhancers that gain H3K27 acetylation after injury are not repressed by H3K27 methylation before injury, which is normally observed in developmentally poised enhancers. Injury induces demethylation of H3K27 in many genes, such as Sonic hedgehog (Shh), which is silenced throughout Schwann cell development before injury. In addition, experiments using a Schwann cell-specific mouse knock-out of the Eed subunit of PRC2 indicate that demethylation is a rate-limiting step in the activation of such genes. We also show that some transcription start sites of H3K27me3-repressed injury genes of uninjured nerves are bound with a mark of active promoters H3K4me3, for example, Shh and Gdnf, and the reduction of H3K27me3 results in increased trimethylation of H3K4. Our findings identify reversal of polycomb repression as a key step in gene activation after injury. Peripheral nerve regeneration after injury is dependent upon implementation of a novel genetic program in Schwann cells that supports axonal survival and regeneration. Identifying means to enhance Schwann cell reprogramming after nerve injury could be used to foster effective remyelination in the treatment of demyelinating disorders and in identifying pathways involved in regenerative process of myelination. Although recent progress has identified transcriptional determinants of successful reprogramming of the Schwann cell transcriptome after nerve injury, our results have highlighted a novel epigenomic pathway in which reversal of the Polycomb pathway of repressive histone methylation is required for activation of a significant number of injury-induced genes.


Magnetoreception in honeybees.
Magnetoreception by honeybees (Apis mellifera) is demonstrated by such activities as comb building and homing orientation, which are affected by the geomagnetic field. In other magnetoreceptive species, iron oxide crystals in the form of magnetite have been shown to be necessary for primary detection of magnetic fields. Here it is shown that trophocytes, which are apparently the only iron granule-containing cells in honeybees, contain super-paramagnetic magnetite. These cells are innervated by the nervous system, which suggests that trophocytes might be primarily responsible for magnetoreception. Electron microscopy also shows cytoskeletal attachments to the iron granule membrane.


Growing burden of noncommunicable diseases: the contributory role of oral diseases, Eastern Mediterranean Region perspective.
Associations of oral diseases with noncommunicable diseases such as diabetes, cardiovascular diseases, chronic respiratory diseases, osteoporosis and chronic renal failure are widely reported in the literature from developed countries. Commonality of risk factors, changes in systemic inflammatory mediators and body metabolism play a role in this association. This paper reviews current knowledge on the burden and association of oral and systemic diseases, and highlights the paucity of information and research from the Eastern Mediterranean Region and other developing countries. A call is made for further research to understand the status and significance of oral-systemic disease associations and develop guidelines for their control in this Region.


Advances in research on fat infiltration and lumbar intervertebral disc degeneration.
Low back pain (LBP) is a disabling condition with no available cure, severely affecting patients' quality of life. Intervertebral disc degeneration (IVDD) is the leading cause of chronic low back pain (CLBP). IVDD is a common and recurrent condition in spine surgery. Disc degeneration is closely associated with intervertebral disc inflammation. The intervertebral disc is an avascular tissue in the human body. Transitioning from hematopoietic bone marrow to bone marrow fat may initiate an inflammatory response as we age, resulting in bone marrow lesions in vertebrae. In addition, the development of LBP is closely associated with spinal stability imbalance. An excellent functional state of paraspinal muscles (PSMs) plays a vital role in maintaining spinal stability. Studies have shown that the diminished function of PSMs is mainly associated with increased fat content, but whether the fat content of PSMs is related to the degree of disc degeneration is still under study. Given the vital role of PSMs lesions in CLBP, it is crucial to elucidate the interaction between PSMs changes and CLBP. Therefore, this article reviews the advances in the relationship and the underlying mechanisms between IVDD and PSMs fatty infiltration in patients with CLBP.


The armed forces medical services response to COVID-19.
The Indian Armed Forces have always responded to the Nation's call and the COVID-19 pandemic response has been no different. On instructions from the Government of India, the Armed Forces Medical Services (AFMS) pitched in right from the initial stages of the epidemic in India as part of a coordinated national response. Be it the execution of medical quarantine for Indian citizens evacuated from China and other COVID affected countries or establishing dedicated and mixed COVID hospitals for its own clientele as well as civilian patients, the AFMS worked in tandem with the national policies. The Armed Forces ensured force preservation and protection of its own troops and families by timely implementation of public health measures, even as it played its designated role in the national strategy. With vision, understanding and clarity, the AFMS continue to lend shoulder to India's response to this global public health challenge.


Lack of molecular genetic divergence between sea-ranched and wild sea trout (Salmo trutta).
The supportive breeding programme for sea trout (Salmo trutta) in the River Dalälven, Sweden, is based on a sea-ranched hatchery stock of local origin that has been kept 'closed' to the immigration of wild genes since the late 1960s (about seven generations). In spite of an apparent potential for substantial uni directional gene flow from sea-ranched to wild (naturally produced) trout, phenotypic differences with a presumed genetic basis have previously been observed between the two 'stocks'. Likewise, two previous studies of allozyme and mitochondrial DNA variation based on a single year of sampling have indicated genetic differentiation. In the present study we used microsatellite and allozyme data collected over four consecutive years, and tested for the existence of overall genetic stock divergence while accounting for temporal heterogeneity. Statistical analyses of allele frequency variation (F-statistics) and multilocus genotypes (assignment tests) revealed that wild and sea-ranched trout were significantly different in three of four years, whereas no overall genetic divergence could be found when temporal heterogeneity among years within stocks was accounted for. On the basis of estimates of effective population size in the two stocks, and of FST between them, we also assessed the level of gene flow from sea-ranched to wild trout to be approximately 80% per generation (with a lower confidence limit of approximately 20%). The results suggest that the reproductive success of hatchery and naturally produced trout may be quite similar in the wild, and that the genetic characteristics of the wild stock are largely determined by introgressed genes from sea-ranched fish.


The influence of the glutamatergic system on cognition in schizophrenia: A systematic review.
Previous literature showing the role of the glutamatergic system on cognition in schizophrenia has been inconclusive. 44 relevant pharmacological, candidate gene and neuroimaging studies were identified through systematic search following PRISMA guidelines. To be included, studies must have observed at least one objective measure of cognitive performance in patients with schizophrenia and either manipulated or measured the glutamatergic system. Of the cognitive domains observed, memory, working memory and executive functions appear to be most influenced by the glutamatergic pathway. In addition, evidence from the literature suggests that presynaptic components synthesis and uptake of glutamate is involved in memory, while postsynaptic signalling appears to be involved in working memory. In addition, it appears that the glutamatergic pathway is particularly involved in cognitive flexibility and learning potential in regards to executive functioning. The glutamatergic system appears to contribute to the cognitive deficits in schizophrenia, whereby different parts of the pathway are associated with different cognitive domains. This review demonstrates the necessity for cognition to be examined by domain as opposed to globally.


Endovascular Thrombectomy for Distal Occlusion Using a Semi-Deployed Stentriever: Report of 2 Cases and Technical Note.
Distal intracranial occlusions can sometimes cause significant neurological deficits. Endovascular thrombectomy in these vessels may improve outcome but carry a higher risk of haemorrhagic complications due to the small calibre and tortuosity of the target vessel. We report two cases of isolated M2/3 artery occlusion causing dense hemiplegia that was successfully treated with stent retrieval thrombectomy. A "semi-deployment technique" of a 3 mm stentriever was employed at the M2/3 bifurcation of the middle cerebral artery. Partial stent unsheathing allowed adequate clot engagement while avoiding excessive tension by the stent metal struts along the tortuous course of a distal vessel. Complete revascularization was achieved after firstpass of the stent retriever without complication, resulting in good clinical outcome in both cases. The described semi-deployment technique reduces the radial and tractional force exerted by the stentreiver on small branches, and may reduce the risk of vessel laceration or dissection in distal vessel thrombectomy.


Spatio-temporal skin strain distributions evoke low variability spike responses in cuneate neurons.
A common method to explore the somatosensory function of the brain is to relate skin stimuli to neurophysiological recordings. However, interaction with the skin involves complex mechanical effects. Variability in mechanically induced spike responses is likely to be due in part to mechanical variability of the transformation of stimuli into spiking patterns in the primary sensors located in the skin. This source of variability greatly hampers detailed investigations of the response of the brain to different types of mechanical stimuli. A novel stimulation technique designed to minimize the uncertainty in the strain distributions induced in the skin was applied to evoke responses in single neurons in the cat. We show that exposure to specific spatio-temporal stimuli induced highly reproducible spike responses in the cells of the cuneate nucleus, which represents the first stage of integration of peripheral inputs to the brain. Using precisely controlled spatio-temporal stimuli, we also show that cuneate neurons, as a whole, were selectively sensitive to the spatial and to the temporal aspects of the stimuli. We conclude that the present skin stimulation technique based on localized differential tractions greatly reduces response variability that is exogenous to the information processing of the brain and hence paves the way for substantially more detailed investigations of the brain's somatosensory system.


Loss-of-function mutations in the maize homeobox gene, knotted1, are defective in shoot meristem maintenance.
The product of the maize homeobox gene, knotted1 (kn1), localizes to the nuclei of cells in shoot meristems, but is absent from portions of the meristem where leaf primordia or floral organs initiate. Recessive mutant alleles of kn1 were obtained by screening for loss of the dominant leaf phenotype in maize. Mutant kn1 alleles carrying nonsense, splicing and frame shift mutations cause severe inflorescence and floral defects. Mutant tassels produce fewer branches and spikelets. Ears are often absent, and when present, are small with few spikelets. In addition, extra carpels form in female florets and ovule tissue proliferates abnormally. Less frequently, extra leaves form in the axils of vegetative leaves. These mutations reveal a role for kn1 in meristem maintenance, particularly as it affects branching and lateral organ formation.


A Profile Hidden Markov Model to investigate the distribution and frequency of LanB-encoding lantibiotic modification genes in the human oral and gut microbiome.
The human microbiota plays a key role in health and disease, and bacteriocins, which are small, bacterially produced, antimicrobial peptides, are likely to have an important function in the stability and dynamics of this community. Here we examined the density and distribution of the subclass I lantibiotic modification protein, LanB, in human oral and stool microbiome datasets using a specially constructed profile Hidden Markov Model (HMM). The model was validated by correctly identifying known lanB genes in the genomes of known bacteriocin producers more effectively than other methods, while being sensitive enough to differentiate between different subclasses of lantibiotic modification proteins. This approach was compared with two existing methods to screen both genomic and metagenomic datasets obtained from the Human Microbiome Project (HMP). Of the methods evaluated, the new profile HMM identified the greatest number of putative LanB proteins in the stool and oral metagenome data while BlastP identified the fewest. In addition, the model identified more LanB proteins than a pre-existing Pfam lanthionine dehydratase model. Searching the gastrointestinal tract subset of the HMP reference genome database with the new HMM identified seven putative subclass I lantibiotic producers, including two members of the Coprobacillus genus. These findings establish custom profile HMMs as a potentially powerful tool in the search for novel bioactive producers with the power to benefit human health, and reinforce the repertoire of apparent bacteriocin-encoding gene clusters that may have been overlooked by culture-dependent mining efforts to date.


Neurological complications of HIV-1-seropositive internal medicine inpatients in Kinshasa, Zaire.
Because little was known about the prevalence of neurological complications of human immunodeficiency virus type 1 (HIV-1) infection in Africa, we conducted a cross-sectional study among consecutive admissions to the internal medicine wards of Mama Yemo Hospital in Kinshasa, Zaire. Of the 196 patients studied, 104 (53%) were HIV-1 seropositive, of whom 50 (48%) had stage 3 and 49 (47%) had stage 4 HIV-1 infection according to the provisional WHO staging criteria for HIV infection. Neuropsychiatric abnormalities were present in 43 (41%) of 104 HIV-1-seropositive patients. Of the HIV-1-seropositive patients, 9 (8.7%; 95% confidence interval, 4-16%) were diagnosed as having possible HIV-1-associated dementia complex, 1 (1%) as having possible HIV-1 myelopathy, and 3 (2.7%) as having possible HIV-1-associated minor cognitive/motor disorder. Definitive diagnoses could not be made because there were no facilities for neuroimaging and neuropathology. Meningitis caused by cryptococcus was diagnosed in six (5.6%) and by Mycobacterium avium in two (2%) of the HIV-1 seropositive patients. Acute onset hemiplegia, believed to be due to stroke, was present in four (4%) of the HIV-1-seropositive patients. The prevalence of other central nervous system opportunistic infections and mass lesions, especially toxoplasmic encephalitis, could not be assessed. In this population of Zairian inpatients, the prevalence of neurological complications of HIV-1 infection was similar to that observed in industrialized countries among patients with advanced HIV disease.


[Analysis of synaptic transmission in rat hippocampal slices using a modified method of potential fixation].
A new approach for the whole-cell recording in slices is suggested as simpler and less expensive one than those known before. Electrophysiological characteristics of excitatory postsynaptic currents are obtained. Two types of glutamate receptors are shown to be involved. The kynurenic acid is proved to exert more prominent effect on the NMDA-receptor mediated current. Biphasic action of adenosine, a wide spread neuromodulator, on synaptic transmission is shown.


[Partial antigenic structure of group H10 flagellins in Escherichia coli and its genetic determination].
The study of flagellar antigens classified with serotype H10 has been made in E. coli belonging to different OK-groups. These antigens have been shown to differ in their partial structure. Group H10 antigens have been found to comprise 7 variants, and their partial structure has been determined. The study has shown that antigenic variants of the same type occur in E. coli belonging to different OK-groups and different variants may occur in strains within the same OK-group. The data obtained in the study of the expression of genes, responsible for the synthesis of flagellin, in the system of isogenic strains suggest that differences in the partial structure between the variants of group H10 antigens are linked with differences between the corresponding alleles of the flagellin-specifying gene. The differentiation of these antigens by means of factor-specific antibodies may prove to be of practical importance for solving the problems of the epidemiology and etiology of diseases caused by E. coli having group H10 flagellar antigens.


Lichenoid dermatitis in three patients with metastatic melanoma treated with anti-PD-1 therapy.
Therapies that activate the immune system through blocking the binding of programmed death ligand 1 (PD-L1) present on tumors and PD-1 (programmed death 1) present on activated immune cells are revolutionizing the care for patients with cancer. These therapies work by inhibiting negative regulators of the immune system, thereby decreasing a tumor's ability to evade the immune system. The side effects of anti-PD-1/PD-L1 therapies are generally mild and as expected are related to autoimmune reactions. Two of the most common side effects of anti-PD-1/PD-L1 therapies are rash and pruritus occurring in approximately 20% of patients. Although the rash is generally recognized to be immune mediated, the exact mechanisms of the rash remain unclear. Herein, we report three cases of lichenoid dermatitis in three patients treated with MK-3475 (anti-PD-1) that were characterized with marked T-cell infiltrates with few PD-1-positive cells. The rashes in all three patients were relatively mild, allowing treatment to continue despite the rashes.


Quality improvement initiative to reduce serious safety events and improve patient safety culture.
Many thousands of patients die every year in the United States as a result of serious and largely preventable safety events or medical errors. Safety events are common in hospitalized children. We conducted a quality improvement initiative to implement cultural and system changes with the goal of reducing serious safety events (SSEs) by 80% within 4 years at our large, urban pediatric hospital. A multidisciplinary SSE reduction team reviewed the safety literature, examined recent SSEs, interviewed internal leaders, and visited other leading organizations. Senior hospital leaders provided oversight, monitored progress, and helped to overcome barriers. Interventions focused on: (1) error prevention; (2) restructuring patient safety governance; (3) a new root cause analysis process and a common cause database; (4) a highly visible lessons learned program; and (5) specific tactical interventions for high-risk areas. Our outcome measures were the rate of SSEs and the change in patient safety culture. SSEs per 10000 adjusted patient-days decreased from a mean of 0.9 at baseline to 0.3 (P < .0001). The days between SSEs increased from a mean of 19.4 at baseline to 55.2 (P < .0001). After a worsening of patient safety culture outcomes in the first year of intervention, significant improvements were observed between 2007 and 2009. Our multifaceted approach was associated with a significant and sustained reduction of SSEs and improvements in patient safety culture. Multisite studies are needed to better understand contextual factors and the significance of specific interventions.


Identification of myosin II as a cripto binding protein and regulator of cripto function in stem cells and tissue regeneration.
Cripto regulates stem cell function in normal and disease contexts via TGFbeta/activin/nodal, PI3K/Akt, MAPK and Wnt signaling. Still, the molecular mechanisms that govern these pleiotropic functions of Cripto remain poorly understood. We performed an unbiased screen for novel Cripto binding proteins using proteomics-based methods, and identified novel proteins including members of myosin II complexes, the actin cytoskeleton, the cellular stress response, and extracellular exosomes. We report that myosin II, and upstream ROCK1/2 activities are required for localization of Cripto to cytoplasm/membrane domains and its subsequent release into the conditioned media fraction of cultured cells. Functionally, we demonstrate that soluble Cripto (one-eyed pinhead in zebrafish) promotes proliferation in mesenchymal stem cells (MSCs) and stem cell-mediated wound healing in the zebrafish caudal fin model of regeneration. Notably, we demonstrate that both Cripto and myosin II inhibitors attenuated regeneration to a similar degree and in a non-additive manner. Taken together, our data present a novel role for myosin II function in regulating subcellular Cripto localization and function in stem cells and an important regulatory mechanism of tissue regeneration. Importantly, these insights may further the development of context-dependent Cripto agonists and antagonists for therapeutic benefit.


Beyond K-complex binary scoring during sleep: probabilistic classification using deep learning.
K-complexes (KCs) are a recognized electroencephalography marker of sensory processing and a defining feature of sleep stage 2. KC frequency and morphology may also be reflective of sleep quality, aging, and a range of sleep and sensory processing deficits. However, manual scoring of K-complexes is impractical, time-consuming, and thus costly and currently not well-standardized. Although automated KC detection methods have been developed, performance and uptake remain limited. The proposed algorithm is based on a deep neural network and Gaussian process, which gives the input waveform a probability of being a KC ranging from 0% to 100%. The algorithm was trained on half a million synthetic KCs derived from manually scored sleep stage 2 KCs from the Montreal Archive of Sleep Study containing 19 healthy young participants. Algorithm performance was subsequently assessed on 700 independent recordings from the Cleveland Family Study using sleep stages 2 and 3 data. The developed algorithm showed an F1 score (a measure of binary classification accuracy) of 0.78 and thus outperforms currently available KC scoring algorithms with F1 = 0.2-0.6. The probabilistic approach also captured expected variability in KC shape and amplitude within individuals and across age groups. An automated probabilistic KC classification is well suited and effective for systematic KC detection for a more in-depth exploration of potential relationships between KCs during sleep and clinical outcomes such as health impacts and daytime symptomatology.


Biogenesis of regulated exocytotic carriers in neuroendocrine cells.
Ca(2+)-triggered exocytosis is a hallmark of neurosecretory granules, but the cellular pathway leading to the assembly of these regulated exocytotic carriers is poorly understood. Here we used the pituitary AtT-20 cell line to study the biogenesis of regulated exocytotic carriers involved in peptide hormone secretion. We show that immature secretory granules (ISGs) freshly budded from the trans-Golgi network (TGN) exhibit characteristics of unregulated exocytotic carriers. During a subsequent maturation period they undergo an important switch to become regulated exocytotic carriers. We have identified a novel sorting pathway responsible for this transition. The SNARE proteins, VAMP4 and synaptotagmin IV (Syt IV), enter ISGs initially but are sorted away during maturation. Sorting is achieved by vesicle budding from the ISGs, because it can be inhibited by brefeldin A (BFA). Inhibition of this sorting pathway with BFA arrested the maturing granules in a state that responded poorly to stimuli, suggesting that the transition to regulated exocytotic carriers requires the removal of a putative inhibitor. In support of this, we found that overexpression of Syt IV reduced the stimulus-responsiveness of maturing granules. We conclude that secretory granules undergo a switch from unregulated to regulated secretory carriers during biogenesis. The existence of such a switch may provide a mechanism for cells to modulate their secretory activities under different physiological conditions.


Dinucleotide Degradation by REXO2 Maintains Promoter Specificity in Mammalian Mitochondria.
Oligoribonucleases are conserved enzymes that degrade short RNA molecules of up to 5 nt in length and are assumed to constitute the final stage of RNA turnover. Here we demonstrate that REXO2 is a specialized dinucleotide-degrading enzyme that shows no preference between RNA and DNA dinucleotide substrates. A heart- and skeletal-muscle-specific knockout mouse displays elevated dinucleotide levels and alterations in gene expression patterns indicative of aberrant dinucleotide-primed transcription initiation. We find that dinucleotides act as potent stimulators of mitochondrial transcription initiation in vitro. Our data demonstrate that increased levels of dinucleotides can be used to initiate transcription, leading to an increase in transcription levels from both mitochondrial promoters and other, nonspecific sequence elements in mitochondrial DNA. Efficient RNA turnover by REXO2 is thus required to maintain promoter specificity and proper regulation of transcription in mammalian mitochondria.


Delays in gender affirming healthcare due to COVID-19 are mitigated by expansion of telemedicine.
Gender-affirming healthcare is vital for transgender and gender diverse (TGD) patients, and during the pandemic, accessing healthcare became challenging. Hypothesizing that many had procedures postponed, we sought to characterize the impact of the pandemic on TGD patients. A mixed-methods approach was employed, combining surveys and interviews; Duke patients were identified by ICD-10 codes, while non-Duke (national) patients were recruited through online social media. All specialties increased telemedicine usage during the pandemic. Duke surgical patients reported a nearly three-fold increase in telemedicine access. COVID-19 symptoms were reported by 24% of Duke and 20% of national patients; barriers to urgent care included the fear of discrimination (27%). Delays were experienced in all domains of care, mitigated in part by telemedicine. Nearly one-third of patients cite discrimination as a barrier to care. Though pandemic-related expansion of telemedicine may be a marker of success, significant barriers still complicate delivery of healthcare.


Physical Activity Assessment Between Consumer- and Research-Grade Accelerometers: A Comparative Study in Free-Living Conditions.
Wearable activity monitors such as Fitbit enable users to track various attributes of their physical activity (PA) over time and have the potential to be used in research to promote and measure PA behavior. However, the measurement accuracy of Fitbit in absolute free-living conditions is largely unknown. To examine the measurement congruence between Fitbit Flex and ActiGraph GT3X for quantifying steps, metabolic equivalent tasks (METs), and proportion of time in sedentary activity and light-, moderate-, and vigorous-intensity PA in healthy adults in free-living conditions. A convenience sample of 19 participants (4 men and 15 women), aged 18-37 years, concurrently wore the Fitbit Flex (wrist) and ActiGraph GT3X (waist) for 1- or 2-week observation periods (n=3 and n=16, respectively) that included self-reported bouts of daily exercise. Data were examined for daily activity, averaged over 14 days and for minutes of reported exercise. Average day-level data included steps, METs, and proportion of time in different intensity levels. Minute-level data included steps, METs, and mean intensity score (0 = sedentary, 3 = vigorous) for overall reported exercise bouts (N=120) and by exercise type (walking, n=16; run or sports, n=44; cardio machine, n=20). Measures of steps were similar between devices for average day- and minute-level observations (all P values > .05). Fitbit significantly overestimated METs for average daily activity, for overall minutes of reported exercise bouts, and for walking and run or sports exercises (mean difference 0.70, 1.80, 3.16, and 2.00 METs, respectively; all P values < .001). For average daily activity, Fitbit significantly underestimated the proportion of time in sedentary and light intensity by 20% and 34%, respectively, and overestimated time by 3% in both moderate and vigorous intensity (all P values < .001). Mean intensity scores were not different for overall minutes of exercise or for run or sports and cardio-machine exercises (all P values > .05). Fitbit Flex provides accurate measures of steps for daily activity and minutes of reported exercise, regardless of exercise type. Although the proportion of time in different intensity levels varied between devices, examining the mean intensity score for minute-level bouts across different exercise types enabled interdevice comparisons that revealed similar measures of exercise intensity. Fitbit Flex is shown to have measurement limitations that may affect its potential utility and validity for measuring PA attributes in free-living conditions.


Disease flare after tyrosine kinase inhibitor discontinuation in patients with EGFR-mutant lung cancer and acquired resistance to erlotinib or gefitinib: implications for clinical trial design.
Treatment of patients with oncogene-addicted cancers with tyrosine kinase inhibitors (TKI) is biologically and clinically different than with cytotoxic chemotherapy. We have observed that some patients with EGFR-mutant lung cancer and acquired resistance to erlotinib or gefitinib (RECIST progression after initial benefit) have accelerated progression of disease after discontinuation of TKI. To examine this observation and define the course of patients following TKI discontinuation, we systematically evaluated patients enrolled on clinical trials of agents to treat acquired resistance to erlotinib or gefitinib. We evaluated patients with EGFR-mutant lung cancer who participated in trials for patients with acquired resistance that mandated TKI discontinuation before administration of study therapy. Disease flare was defined as hospitalization or death attributable to disease progression during the washout period. Fourteen of 61 patients (23%; 95% CI: 14-35) experienced a disease flare. The median time to disease flare after TKI discontinuation was 8 days (range 3-21). Factors associated with disease flare included shorter time to progression on initial TKI (P = 0.002) and the presence of pleural (P = 0.03) or CNS disease (P = 0.01). There was no association between disease flare and the presence of T790M at the time of acquired resistance. In patients with EGFR-mutant lung cancer and acquired resistance to epidermal growth factor receptor TKIs, discontinuation of erlotinib or gefitinib before initiation of study treatment is associated with a clinically significant risk of accelerated disease progression. Clinical trials in this patient population must minimize protocol-mandated washout periods.