Growth charts in FGFR2- and FGFR3-related faciocraniosynostoses.
Faciocraniosynostoses (FCS) are malformations affecting the development of the bones of the skull and face, due to the premature closure of one or more craniofacial sutures, mostly secondary to activating Fibroblast Growth Factor Receptor (FGFR) 1-3 mutations. Gain-of-function FGFR3 mutations are also responsible for various conditions referred to as osteochondrodysplasia (OCD), characterized by structural and functional abnormalities of growth plate cartilages. We hypothesized that patients with FGFR-related faciocraniosynostoses may present extra-cranial growth anomalies. We retrospectively collected height and weight data from a cohort of 70 patients. Included patients were admitted for FGFR-related FCS between 2000 and 2021 at the Craniofacial Unit of Necker - Enfants Malades University Hospital in Paris, France. We showed that FGFR-related faciocraniosynostoses had significantly reduced heights and weights relative to controls, and that two specific time periods (1-3 years and > 8 years of age) were associated with lower height and weight values. Four patients had received growth hormone treatment but remained below normal values for growth in height and weight. Patients with FGFR-related faciocraniosynostoses have clinically significant extra-cranial anomalies which are not currently investigated and managed in usual protocols; these patients could benefit from a systematic pre-pubertal endocrine assessment. More generally, our results extend the scope of extracranial anomalies in FGFR-related faciocraniosynostoses and support the hypothesis that all conditions with activating FGFR mutations affect both membranous ossification and long bones.


Methyl 2-[2-(tert-but-oxy-carbonyl-amino)-1,3-benzothia-zole-6-carboxamido]-acetate.
In the title compound, C(16)H(19)N(3)O(5)S, the dihedral angle between the benzene ring and the carbonyl-amino group is 18.18 (2)°. In the crystal, mol-ecules form centrosymmetric dimers via pairs of N-H⋯N hydrogen bonds. The dimers are connected via N-H⋯O hydrogen bonds into a three-dimensional network..


Nutrient cycling by the subterranean termite Gnathamitermes tubiformans in a Chihuahuan desert ecosystem.
We estimated the density of subterranean termites Gnathamitermes tubiformans at 800,000 · ha-1 for a standing crop biomass of 2 kg · ha-1 Predation losses were estimated to be 5,73 kg · ha-1 · yr-1 representing the major release of nutrients from termites to surficial soil layers. Nutrient fluxes from termites to predators amounted to 410g N·ha-1·yr-1, 33 g S · ha-1 · yr-1 and 19 g P · ha-1 · yr-1. These fluxes account for 8% of the litter N, 1.5% of the litter P and 2.9% of the litter S. The termites fixed an estimated 66 g · ha-1 · yr-1 atmospheric N and returned an estimated 100 g · ha-1 · yr-1 in the surface gallery carton. Since losses of elements from subterannean termites were greater than standing crops, we estimated an annual turnover of N at 3.5 times per year, P of 2.5 times per year, and S of 2.5 per times per year.Since surface foraging, predation and alate flights are pulse regulated by rainfall, nutrient flows through subterranean termites are episodic and releases of nutrients accumulated in termite biomass preceeds or is coincident with productivity "pulses" of some shallow rooted plants. We propose that subterranean termites are important as regulators in desert nutrient cycles.


Harmful health effects of pesticides used on museum textile artifacts - overview.
The paper discusses the types of fibers contained in the museum textile artifacts and the types of pests and pesticides. Also, the main health problems associated with pesticide exposure are presented. Textile fibers represent an excellent source of food for microbes and insects. The key to the long-term preservation of textiles is by implementing an integrated management program for pest and environmental conditions control. The main types of pesticides are insecticides, herbicides, fungicides, rodenticides, acaricides, and fumigants and a way for classification can be made according to the LD50 (lethal dose) value which evaluates the toxicity of a pesticide. Depending on the LD50 values, pesticides can be very toxic, moderately toxic, slightly toxic, and relatively non-toxic. Exposure to pesticides can occur through 4 four routes: oral, dermal, respiratory, and eye exposure. The main diseases that can be associated with exposure to pesticides are discussed in the paper. In the end, a basic set of precautionary recommendations when handling museum objects that may contain hazardous materials was presented.


Regional variation in expression of pro-inflammatory mediators in the intestine following a combined insult of alcohol and burn injury.
The intestine is segmented into functionally discrete compartments (duodenum, jejunum, ileum, and colon). The present study examined whether alcohol combined with burn injury differently influences cytokine levels in different parts of the intestine. Male mice were gavaged with alcohol (∼2.9 g/kg) 4 h prior to receiving a ∼12.5% total body surface area full thickness burn. Mice were sacrificed 1, 3, and 7 days after injury. The intestine segments (duodenum, jejunum, ileum, and colon) were harvested, homogenized, and analyzed for inflammatory mediators (IL-6, IL-18, and KC) using their respective ELISAs. KC levels were significantly increased in the jejunum, ileum, and colon following alcohol and burn injury as compared to shams. The increase in KC was ∼28-fold higher in the colon as compared to the levels observed in duodenum following alcohol and burn injury. Both IL-6 and IL-18 levels were significantly elevated in both the ileum and colon following the combined insult. There was a ∼7-fold increase in IL-6 levels in the colon as compared with the duodenum after the combined insult. Levels of IL-18 were increased by ∼1.5-fold in the colon as compared to the ileum following alcohol and burn injury. The data suggest that pro-inflammatory mediators are differentially expressed in the intestine following alcohol and burn injury.


Regulation of the salvage pathway of purine nucleotide synthesis by the oxidation state of NAD+ in rat heart cells.
The rate of salvage of purine nucleotides from hypoxanthine in glycolyzing, cultured rat heart cells was found to be decreased when respiration was suppressed. Pyruvate or phenazine methosulfate, acting as hydrogen acceptors, reversed the effect of the respiratory block. The inhibition and the reversal could not be attributed to the limitation of energy supply or of 5-phosphoribosyl-1-pyrophosphate. A causal connection was, however, shown to exist between this inhibition and the concomitant shift in the redox state of NAD+ in favor of NADH. NADH also inhibited the key enzyme of the salvage pathway, hypoxanthine-guanine-phosphoribosyltransferase, in cell-free extracts. Regulation of purine nucleotide synthesis by the redox state of NAD+ in heart cells might gain significance during transition from respiring to hypoxic state and vice versa.


Spatial control of photomechanical movements in the lateral eye of the American horseshoe crab, Limulus polyphemus.
This study asks whether photomechanical movements in the retinal cells of the lateral eye of the American horseshoe crab, Limulus polyphemus, are controlled locally within each ommatidium, or whether they are a retinal array property involving lateral communication between ommatidia. Three experiments were performed. A small spot, a vertical slit down the center, or the anterior third of an otherwise masked eye was illuminated. The contralateral eye was fully illuminated in each experiment and served as a light-adapted control. Morphometric analyses of aperture length and rhabdom dimensions were made from serial 1-microm plastic sections. The results suggest there is a different spatial threshold for photomechanical movement for aperture lengthening than for rhabdom lengthening. When only a few ommatidia are illuminated, the aperture does not change. When about 10% are illuminated, they lengthen, but the masked ommatidia do not. When about a third are illuminated, all the ommatidia in the eye lengthen together, including the two thirds that were masked. When either only a few ommatidia or about 10% of the ommatidia are illuminated, rhabdom shape is unchanged. When a third of the eye is illuminated, the illuminated rhabdoms lengthen, but the masked rhabdoms do not.


Periorbital cellulitis.
Periorbital cellulitis is a common complication of sinusitis in children. At some hospitals lumbar puncture is routinely done as part of the complete evaluation of children admitted with periorbital cellulitis. Presumably, the lumbar puncture is done to rule out meningitis which could be the result of extension of infection from the orbit to the central nervous system (CNS). However, the degree to which periorbital cellulitis is associated with CNS complication is not known and the necessity for routine lumbar puncture is questioned. Therefore, records of 102 children admitted to the Children's Hospital National Medical Center from 1975 to 1982 with the diagnosis of periorbital cellulitis were reviewed. There were two cases of meningitis and one case of bilateral subdural effusions. Review of the literature and the findings in this study suggests that routine lumbar puncture in seemingly uncomplicated cases of periorbital cellulitis probably is not indicated in children 6 months of age or older.


Human neutrophils contain distinct cytosolic and particulate tyrosine kinase activities: possible role in neutrophil activation.
Tyrosine protein kinase activities were partially purified from circulating human neutrophils. Purification steps involved sequential chromatography on DEAE-Sephacel, gel filtration and affinity chromatography on a column composed of a glutamine:tyrosine copolymer linked to AH-Sepharose. The results indicate that human neutrophils contain a tyrosine kinase activity in the 150,000 x g cytosolic fraction which is distinct from the activity in a detergent extractable 150,000 x g particulate fraction. These enzyme activities are dependent on the divalent cations Mn2+ and Mg2+. Kinetics for the phosphorylation of a glutamine:tyrosine copolymer substrate demonstrated an apparent Km for the cytosolic tyrosine kinase activity of 22.3 +/- 0.3 microM, and an apparent Km for the particulate extract activity of 42.7 +/- 6.0 microM. By gel filtration chromatography, the cytosolic and particulate tyrosine kinase activities have approximate molecular masses of 80-90 and 50-60 kDa, respectively. The particulate but not the cytosolic neutrophil tyrosine kinase activity was inhibited by a novel tyrosine kinase inhibitor ST638. ST638 inhibited superoxide production in intact neutrophils stimulated with the chemotactic peptide F-Met-Leu-Phe, opsonized zymosan particles, and sodium fluoride. ST638 did not, however, inhibit superoxide production in neutrophils stimulated with phorbol myristate acetate or the calcium ionophore, A23187.


Molecular diagnosis of childhood tuberculosis and infection with Bacilli Calmette-Guerin in Taiwan.
Molecular techniques along with clinical evaluation have been demonstrated to be effective for differentiating childhood tuberculosis (TB), and for establishing an enhanced survey of adverse reactions of Bacilli Calmette-Guerin vaccination in Taiwan. Future development and evaluation of new diagnostics should be prioritized in strengthening the management of childhood TB.


Longitudinal Association of Changes in Parental Correlates With Screen Time in Chinese Preschoolers.
This study aimed to explore the relationship between the changes in parent-related factors and preschoolers exceeding screen time (ST) recommendations. A longitudinal analysis using 2-year follow-up data from 4 kindergartens (n = 409) was conducted in Zhejiang, China, from 2019 to 2021. Multivariate logistic regression models were used to determine the potential parental modifiable predictors. The significant associations of baseline ST, change in screen accessibility, and the interaction of preschooler ST with maternal ST change with preschooler follow-up ST were observed. For preschool-aged children with baseline ST ≤ 1 hour per day, the follow-up of preschoolers with ST > 1 hour per day increased significantly when parental clarity of their ST rules decreased or remained low. For preschool children with baseline ST > 1 hour per day, follow-up ST increased significantly when their father kept ST >2 hours per day, when the screen accessibility became or remained easy, or when parental awareness of the ST decreased. Changes in parental correlates played an important role in preschooler ST based on 2-year longitudinal data. Early interventions should focus on improving the clarity of parental rules and perceptions, as well as on reducing parental ST and accessibility of home screens.


Nutritional Management of Short Bowel Syndrome.
Short bowel syndrome (SBS) of infancy is a cause of prolonged morbidity with intolerance to enteral feeding, specialized nutritional needs, and partial/total dependence on parenteral nutrition. These infants can benefit from individualized nutritional strategies to support and enhance the process of intestinal adaptation. Early introduction of enteral feeds during the period of intestinal adaptation is crucial, even though the enteral feedings may need to be supplemented with an effective, safe, and nutritionally adequate parenteral nutritional regimen. Newer generation intravenous lipid emulsions can be effective in preventing and treating intestinal failure-associated liver disease. Prevention of infection(s), pharmaceutical interventions to enhance bowel motility and prevent/mitigate bacteria overgrowth, and specialized multidisciplinary care to minimize the injury to other organs such as the liver, kidneys, and the brain can assist in nutritional rehabilitation and lower the morbidity in SBS.


Facet Inclinations and Interfacet Angle of the Distal Radius on Posteroanterior Radiographs: Clinical Associations With 3 Carpal Pathologies.
The radiographic interfacet angle (IFA), scaphoid facet inclination (SFI), and lunate facet inclination (LFI) of the distal radius were measured in patients with 3 distinct wrist pathologies to determine whether there is an association between these radiographic measurements and these conditions. Posteroanterior wrist radiographs were compiled from patients with 3 types of common wrist pathologies (scaphoid waist fracture [n = 54], scapholunate [SL] dissociation [n = 23], and dorsal ganglion [n = 51]). The patients were all Caucasians aged 20 to 45 years who met strict radiographic criteria. The IFA, SFI, and LFI values of these patients were compared with those obtained from 400 normal wrist radiographs of subjects who met the same selection criteria. In men with a scaphoid waist fracture, the IFA and SFI were significantly greater than in normal men, whereas the LFI was significantly lower. In the SL dissociation group, for all patients and for subgroups stratified according to sex, the IFA and SFI were significantly lower than in the normal matched groups. In the dorsal ganglion group, differences were found in the IFA and SFI for women, but not for men. The facet orientations of the distal radius in patients with scaphoid fracture, SL dissociation, and dorsal ganglion differed from those in the normal population. The IFA alone is most likely to be associated with all 3 pathologies. The SFI and LFI are less likely to be associated with patients with carpal pathologies. Patients with a greater IFA may be susceptible to scaphoid fractures when they fall on an overstretched hand. Patients with a smaller IFA may be susceptible to SL dissociation when they fall on an overstretched hand.


Transepithelial riboflavin/ultraviolet. a corneal cross-linking in keratoconus: morphologic studies on human corneas.
To evaluate histologic and molecular changes in human keratoconic corneas after the procedure of transepithelial collagen cross-linking (CXL), without the removal of corneal epithelium. Experimental laboratory investigation. Thirty corneal buttons were examined, 18 of which were from patients affected by severe keratoconus and submitted to penetrating keratoplasty (PK). Among these, 8 were analyzed without any treatment, 4 were treated with transepithelial CXL 2 hours before PK, and 6 were treated with transepithelial CXL 3 months before PK. Twelve normal corneal buttons from healthy donors were used as controls. The corneal buttons were then evaluated by hematoxylin-eosin staining and by immunostaining with markers of epithelial junction proteins (ß-catenin and connexin 43), of stromal keratocytes (CD34), of apoptosis (terminal deoxynucleotidyl transferase dUTP nick end labeling [TUNEL] assay), and of collagen type I fibers. The analysis of epithelial markers showed a clear defective expression in keratoconic corneas before and soon after the transepithelial CXL treatment, returning to normal in corneas analyzed 3 months after transepithelial CXL. The analysis of stroma components indicated a loss of keratocytes in the upper stroma of keratoconic corneas and a trend toward a normal situation 3 months after transepithelial CXL; similarly, collagen fibers appeared disorganized in keratoconus, while their pattern appears to be close to normal 3 months after treatment. Histologic and immunohistochemical findings on human keratoconic corneas showed the presence of biochemical and morphologic alterations in the epithelium and the upper stroma that are significantly improved 3 months after transepithelial CXL. However, further studies are necessary to assess to what extent these results correlate with measurable biomechanical effects.


Recurrent appendicitis following successful drainage of appendicular abscess in adult without interval appendectomy during COVID-19. Prospective cohort study.
COVID-19 infection is a global pandemic that affected routine health services and made patients fear to consult for medical health problems, even acute abdominal pain. Subsequently, the incidence of complicated appendicitis increased during the Covid-19 pandemic. This study aimed to evaluate recurrent appendicitis after successful drainage of appendicular abscess during COVID-19. A prospective cohort study conducted in the surgical emergency units of our Universities' Hospitals between March 15, 2020 to August 15, 2020 including patients who were admitted with the diagnosis of an appendicular abscess and who underwent open or radiological drainage. Main outcomes included incidence, severity, and risk factors of recurrent appendicitis in patients without interval appendectomy. A total of 316 patients were included for analysis. The mean age of the patients was 37 years (SD ± 13). About two-thirds of patients were males (60.1%). More than one-third (39.6%) had co-morbidities; type 2 diabetes mellitus (T2DM) (22.5%) and hypertension (17.1%) were the most frequent. Approximately one quarter (25.6%) had confirmed COVID 19 infection. About one-third of the patients (30.4%) had recurrent appendicitis. More than half of them (56.3%) showed recurrence after three months, and 43.8% of patients showed recurrence in the first three months. The most frequent grade was grade I (63.5%). Most patients (77.1%) underwent open surgery. Age, T2DM, hypertension, COVID-19 infection and abscess size >3 cm were significantly risking predictors for recurrent appendicitis. Interval appendectomy is suggested to prevent 56.3% of recurrent appendicitis that occurs after 3 months. We recommend performing interval appendectomy in older age, people with diabetes, COVID-19 infected, and abscesses more than 3 cm in diameter. Is interval appendectomy preventing a high incidence of recurrent appendicitis after successful drainage of appendicular abscess during COVID-19 pandemic?


Comparison of the p125 coding region of bovine viral diarrhea viruses.
The p125 (p54/p80) coding region of two cytopathic (CP) strains (Oregon and Singer) and two noncytopathic (NCP) strains (NY-1 and Draper) of bovine viral diarrhea virus (BVDV) were amplified by the polymerase chain reaction, cloned and sequenced. The sequence data confirmed that the two CP strains do not possess any insertion or deletion in their p125 gene as observed in many other CP strains. In the p80, which showed a high amino acid sequence homology among all strains, no amino acid substitution should could be found which distinguished these CP strains from the NCP strains NY-1 and SD-1. Many amino acid substitutions were found in p54 but their individual importance in the CP phenotype is not clear since critical domains of p54 have not yet been experimentally defined. The p54 protein is much less conserved than p80, and sequence homology, as well as dendrogram analysis, permitted us to distinguish two genotypic groups of BVDV (Ia and Ib). The mean homology between strains of these two groups was 77.3/80.4% for the nucleic acid/amino acid sequences while it was 88.0/88.8% and 91.6/93.3% within groups Ia and Ib, respectively. Furthermore, we found that the p125 sequence of our NY-1 strain showed only 92% sequence homology with the partial p80 gene reported for NY-1 but 99.8% homology with another partial sequence of the p125 gene of NY-1 reported elsewhere. These observations underscored the difficulty of maintaining a specific BVDV strain, especially the NCP biotype, in cell cultures.


From monitoring to action: utilising health survey data in national policy development and implementation in Finland.
Health interview and examination surveys provide valuable information for policy, practice and research purposes. Appropriate use of high-quality, representative and timely population data can indirectly help the citizens to live healthier and longer lives. The aim of this study was to review how health survey data have supported health policy making, health research and everyday health care in Finland. Data were collected by focused interviews with ten Finnish senior experts from the Ministry of Social Affairs and Health, political parties, National Institute for Health and Welfare, universities, and health associations. Most interviewees agreed that health surveys have positively affected the health of the population over the past 50 years - through health strategies, care guidelines, legislation, research, prevention programs, risk calculators, and healthier products on the market. There is also a need for further development: the latest research results should be provided in a nutshell for politicians, and effective tools should be developed more for health care professionals' use. The coverage of health information on children, adolescents, oldest old, disabled persons, migrants and ethnic minorities should be improved. Sound health policy and its successful implementation require extensive national cooperation and new communication strategies between policy makers, researchers, health care professionals, health service providers - and citizens. The future health information system in Finland should better cover all population groups. To obtain more comprehensive health information, the possibilities for register linkages should be secured and register data should be further evaluated and developed to serve health monitoring purposes.


Variation of Burkholderia cenocepacia cell wall morphology and mechanical properties during cystic fibrosis lung infection, assessed by atomic force microscopy.
The influence that Burkholderia cenocepacia adaptive evolution during long-term infection in cystic fibrosis (CF) patients has on cell wall morphology and mechanical properties is poorly understood despite their crucial role in cell physiology, persistent infection and pathogenesis. Cell wall morphology and physical properties of three B. cenocepacia isolates collected from a CF patient over a period of 3.5 years were compared using atomic force microscopy (AFM). These serial clonal variants include the first isolate retrieved from the patient and two late isolates obtained after three years of infection and before the patient's death with cepacia syndrome. A consistent and progressive decrease of cell height and a cell shape evolution during infection, from the typical rods to morphology closer to cocci, were observed. The images of cells grown in biofilms showed an identical cell size reduction pattern. Additionally, the apparent elasticity modulus significantly decreases from the early isolate to the last clonal variant retrieved from the patient but the intermediary highly antibiotic resistant clonal isolate showed the highest elasticity values. Concerning the adhesion of bacteria surface to the AFM tip, the first isolate was found to adhere better than the late isolates whose lipopolysaccharide (LPS) structure loss the O-antigen (OAg) during CF infection. The OAg is known to influence Gram-negative bacteria adhesion and be an important factor in B. cenocepacia adaptation to chronic infection. Results reinforce the concept of the occurrence of phenotypic heterogeneity and adaptive evolution, also at the level of cell size, form, envelope topography and physical properties during long-term infection.


Ordered DNA release and target capture in RAG transposition.
Following V(D)J cleavage, the newly liberated DNA signal ends can be either fused together into a signal joint or used as donor DNA in RAG-mediated transposition. We find that both V(D)J cleavage and release of flanking coding DNA occur before the target capture step of transposition can proceed; no coding DNA is ever detected in the target capture complex. Separately from its role in V(D)J cleavage, the DDE motif of the RAG1/2 active site is specifically required for target DNA capture. The requirement for cleavage and release of coding DNA prior to either physical target binding or functional target commitment suggests that the RAG1/2 transposase contains a single binding site for non-RSS DNA that can accommodate either target DNA or coding DNA, but not both together. Perhaps the presence of coding DNA may aid in preventing transpositional resolution of V(D)J recombination intermediates.


Union, complication, reintervention and failure rates of surgical techniques for large diaphyseal defects: a systematic review and meta-analysis.
To understand the potential and limitations of the different available surgical techniques used to treat large, long-bone diaphyseal defects by focusing on union, complication, re-intervention, and failure rates, summarizing the pros and cons of each technique. A literature search was performed on PubMed, Web of Science, and Cochrane databases up to March 16th, 2022; Inclusion criteria were clinical studies written in English, of any level of evidence, with more than five patients, describing the treatment of diaphyseal bone defects. The primary outcome was the analysis of results in terms of primary union, complication, reintervention, and failure rate of the four major groups of techniques: bone allograft and autograft, bone transport, vascularized and non-vascularized fibular graft, and endoprosthesis. The statistical analysis was carried out according to Neyeloff et al., and the Mantel-Haenszel method was used to provide pooled rates across the studies. The influence of the various techniques on union rates, complication rates, and reintervention rates was assessed by a z test on the pooled rates with their corresponding 95% CIs. Assessment of risk of bias and quality of evidence was based on Downs and Black's "Checklist for Measuring Quality" and Rob 2.0 tool. Certainty of yielded evidence was evaluated with the GRADE system. Seventy-four articles were included on 1781 patients treated for the reconstruction of diaphyseal bone defects, 1496 cases in the inferior limb, and 285 in the upper limb, with trauma being the main cause of bone defect. The meta-analysis identified different outcomes in terms of results and risks. Primary union, complications, and reinterventions were 75%, 26% and 23% for bone allografts and autografts, 91%, 62% and 19% for the bone transport group, and 78%, 38% and 23% for fibular grafts; mean time to union was between 7.8 and 8.9 months in all these groups. Results varied according to the different aetiologies, endoprosthesis was the best solution for tumour, although with a 22% failure rate, while trauma presented a more composite outcome, with fibular grafts providing a faster time to union (6.9 months), while cancellous and cortical-cancellous grafts caused less complications, reinterventions, and failures. The literature about this topic has overall limited quality. However, important conclusions can be made: Many options are available to treat critical-size defects of the diaphysis, but no one appears to be an optimal solution in terms of a safe, satisfactory, and long-lasting outcome. Regardless of the bone defect cause, bone transport techniques showed a better primary union rate, but bone allograft and autograft had fewer complication, reintervention, and failure rates than the other techniques. The specific lesion aetiology represents a critical aspect influencing potential and limitations and therefore the choice of the most suitable technique to address the challenging large diaphyseal defects.


Reduced vision as a presenting feature of rhombencephalosynapsis.
Rhombencephalosynapsis is a rare congenital abnormality of the cerebellum; there is limited literature of neuro-ophthalmologic features associated with this disease. We report a case identified following the diagnosis of optic nerve hypoplasia.


Structural basis for replicase polyprotein cleavage and substrate specificity of main protease from SARS-CoV-2.
The main protease (Mpro) of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a key enzyme, which extensively digests CoV replicase polyproteins essential for viral replication and transcription, making it an attractive target for antiviral drug development. However, the molecular mechanism of how Mpro of SARS-CoV-2 digests replicase polyproteins, releasing the nonstructural proteins (nsps), and its substrate specificity remain largely unknown. Here, we determine the high-resolution structures of SARS-CoV-2 Mpro in its resting state, precleavage state, and postcleavage state, constituting a full cycle of substrate cleavage. The structures show the delicate conformational changes that occur during polyprotein processing. Further, we solve the structures of the SARS-CoV-2 Mpro mutant (H41A) in complex with six native cleavage substrates from replicase polyproteins, and demonstrate that SARS-CoV-2 Mpro can recognize sequences as long as 10 residues but only have special selectivity for four subsites. These structural data provide a basis to develop potent new inhibitors against SARS-CoV-2.


Prostanoid synthesis and vascular responses to exogenous arachidonic acid following cerebral ischemia in piglets.
In newborn pigs, cerebral ischemia abolishes both increased cerebral prostanoid production and cerebral vasodilation in response to hypercapnia and hypotension. Attenuation of prostaglandin endoperoxide synthase activity could account for the failure to increase prostanoid synthesis and loss of responses to these stimuli. To test this possibility, arachidonic acid (3, 6, or 30 micrograms/ml) was placed under cranial windows in newborn pigs that had been exposed to 20 min of cerebral ischemia. The conversion to prostanoids and pial arteriolar responses to the arachidonic acid were measured. At all three concentrations, arachidonic acid caused similar increases in pial arteriolar diameter in sham control piglets and piglets 1 hr postischemia. Topical arachidonic acid caused dose-dependent increases of PGE2 in cortical periarachnoid cerebral spinal fluid. 6-keto-PGF1 alpha and TXB2 only increased at the highest concentration of arachidonic acid (30 micrograms/ml). Cerebral ischemia did not decrease the conversion of any concentration of arachidonic acid to PGE2, 6-keto-PGF1 alpha, or TXB2. We conclude that ischemia and subsequent reperfusion do not result in inhibition of prostaglandin endoperoxide synthase in the newborn pig brain. Therefore, the mechanism for the impaired prostanoid production in response to hypercapnia and hypotension following cerebral ischemia appears to involve reduction in release of free arachidonic acid.


Psychologists' Contributions to Patient-Centered Medical Homes.
Mounting evidence supports the value of integrated healthcare and the need for interprofessional practice within patient-centered medical homes (PCMH). Incorporating behavioral health services is key to fully implementing the PCMH concept. Unfortunately, psychologists have not been front and center in this integrative and interprofessional care movement nor have they typically received adequate training or experience to work effectively in these integrated care programs. This article builds the case for the value of PCMHs, particularly those that incorporate behavioral health services. Attention is paid to the diverse roles psychologists play in these settings, including as direct service providers, consultants, teachers/supervisors, scholars/program evaluators, and leaders. There is a discussion of the competencies psychologists must possess to play these roles effectively. Future directions are discussed, with a focus on ways psychologists can bolster the PCMH model by engaging in interprofessional partnerships related to education and training, practice, research, and leadership.


Evaluation of Immunosuppressive Activity of Demethylzeylasteral in a Beagle Dog Kidney Transplantation Model.
Several monomers isolated from Tripterygium wilfordii Hook f. (Celastraceae) have attracted worldwide interest. In this study, we established a simple and reliable kidney transplantation model in beagle dog to evaluate the immunosuppressive activity of demethylzeylasteral (T-96), an immunosuppressive monomer isolated from the root xylem of T. wilfordii. Recipient and donor male beagle dogs were obtained from two different breeders to ensure MHC mismatching. All dogs were randomly divided into six groups following kidney transplantation, and different doses of T-96 or cyclosporine A (CsA) were administered to each group during 14 days of observation. The results showed that T-96 alone at a dosage of 10 or 20 mg/kg/day prolonged graft survival up to 10.83 ± 1.47 or 11.17 ± 1.47 days. A combination of T-96 and CsA significantly prolonged the survival time to 13.33 ± 1.75 days. The results demonstrated that T-96 can inhibit acute rejection in kidney transplantation, and the inhibitory effect of T-96 was enhanced when combined with CsA, which suggests the possible use in organ transplantation to prevent immune rejection.


A systematic review and meta-analysis of the prevalence of sleep problems in children with cerebral palsy: how do children with cerebral palsy differ from each other and from typically developing children?
Up to 85% of children with neurodevelopmental disorders have sleep problems, compared with 25% of typically developing children. Children with cerebral palsy (CP)may have risk factors (brain injury, physical disability, and comorbidities) that make them more likely to have sleep problems compared with typically developing children. To determine prevalence of sleep problems in children with CP. We conducted a systematic review and meta-analysis to report on the prevalence of sleep problems in children with CP, within subgroups (age, CP phenotype, presence of impairments [auditory, visual, and cognitive], and presence of epilepsy) and compared with control groups of healthy children. We searched eight relevant electronic databases from their respective start dates until September 2018. 23 full-text articles (n=2,908 children with CP) were included in the review. All studies were cross-sectional and examined caregiver-reported sleep measures. The Sleep Disturbance Scale for Children (SDSC) was the most commonly used questionnaire. No study met all Joanna Briggs Institute quality assessment criteria for prevalence studies; selection, coverage, classification, and/or confounding biases were present in all studies. Using a random effects model with a Freeman-Tukey double arcsine transformation, the pooled prevalence was 23.4% (95% confidence interval [CI] 18.8-28.4%; n=9 studies) for an abnormal total score on the SDSC and 26.9% (95% CI 21.5-32.7%; n=9 studies) for disorders of initiation and maintenance of sleep, the most prevalent sleep problem reported. For the studies that reported prevalence for control groups of healthy children (n=4 studies), sleep problems were generally more prevalent in the CP group. The prevalence of sleep problems in children with CP is high. There is notable variability in the prevalence of sleep problems between subgroups of children with CP. Future studies using questionnaires validated in children with CP and objective measures (such as polysomnography or actigraphy) in well-described, large, broadly recruited samples are recommended.


Online oxygen measurements in ex vivo perfused muscle tissue in a porcine model using dynamic quenching methods.
Transplantation of autologous free tissue flaps is the best applicable technique for treating large and complex tissue defects and still has one major failure criterion. Tissue--and in particular muscle tissue--is strongly sensitive to ischemia, thus after a critical period of oxygen depletion the risk of a partial or total flap loss is high. For that reason a miniaturized ex vivo perfusion system has been developed, that supplies the tissue during operational delays. The purpose of this study was to determine the oxygenation levels during such a perfusion using different perfusates and therefore to objectify if a complementary oxygenation unit is required to improve perfusion quality. The oxygen levels of the tissue, as well of the perfusate, were measured by using minimal invasive optical oxygen sensors that are based on dynamic quenching. The ex vivo perfused tissue was the porcine rectus abdominis muscle. Results show, that during perfusion with heparinized crystalloid fluid (Jonosteril) and heparinized autologous whole blood, additional oxygenation of the perfusion reactor led to different ex vivo oxygen tissue saturations, which can be detected by dynamic quenching. Dynamic quenching methods are a promising and valuable technique to perform online oxygen measurements in ex vivo perfused muscle tissue in a porcine model.


Novel metal-binding proteins by design.
We describe the successful design of a tetrahedral His3Cys Zn(II)-binding site in a small protein of known structure: the B1 domain of Streptococcal protein G. The B1 variants containing the novel metal-binding site were characterized using a combination of optical absorption, circular dichroism and NMR spectroscopies. The results indicate that the designed proteins bind Zn(II) with high affinity and tetrahedral coordination geometry, and that the overall secondary and tertiary structure of the B1 domain is maintained.


Association between vitamin D metabolism gene polymorphisms and risk of islet autoimmunity and progression to type 1 diabetes: the diabetes autoimmunity study in the young (DAISY).
Vitamin D metabolism genes have been associated with type 1 diabetes (T1D) risk; however, these genes have not been investigated for association with the preclinical phase of T1D, islet autoimmunity (IA). Studies of vitamin D metabolism genes may elucidate the role of vitamin D in complex diseases. The objective of the study was to explore the association between seven vitamin D metabolism gene single-nucleotide polymorphisms (SNPs) and the risk of IA and progression to T1D. The Diabetes Autoimmunity Study in the Young is a longitudinal, observational study. Newborn screening for human leukocyte antigen, sibling and offspring recruitment, and follow-up took place in Denver, Colorado. A total of 1708 children at increased genetic risk of T1D participated in the study: 148 developed IA and 62 IA-positive children progressed to T1D. IA, defined as positivity for glutamic acid decarboxylase, insulin, or IA-2 autoantibodies on two or more consecutive visits, and T1D, diagnosed by a physician, were the main outcome measures. The risk of IA was associated with DHCR7/NADSYN1 rs12785878 and CYP27B1 rs4646536 [hazard ratio 1.36, 95% confidence interval 1.08-1.73 (for each additional minor allele) and hazard ratio 0.59, 95% confidence interval 0.39-0.89 (for A/G compared with the A/A genotype), respectively]. None of the vitamin D SNPs typed was associated with progression to T1D in IA-positive children. Six of the seven SNPs were significantly associated with 25-hydroxyvitamin D levels. DHCR7/NADSYN1 rs12785878 and CYP27B1 rs4646536 may play an important role in islet autoimmunity, the preclinical phase of T1D. These findings should be replicated in larger cohorts for confirmation.


[Disturbances of fusion after extensive retinal haemorrhages in new-born infants (author's transl)].
Follow-up examinations of 21 non-squinting children aged six revealed, that most children who had had extensive bilateral post-partum retinal hemorrhages had a significantly higher incidence of impaired range of fusion (25 degrees max.) as compared to the cases without perinatal hemorrhages. Not a single case of impaired fusion could be improved by orthoptic treatment. The disorder is assumed to be due to a discrete perinatal impairment of the cerebral optomotor centers.


Enteroviruses in recreational waters of Northern Ireland.
Virus surveillance of Northern Ireland recreational waters, between April 1986 and May 1989 demonstrated widespread enteroviral contamination of coastal and inland waters. In 1986, enteroviruses were detected in 4 of 46 (8.7%) water samples, collected from 6 coastal bathing waters. In 1987, 49 of 107 (45.8%) samples, from 16 coastal bathing waters, yielded enteroviruses; 33 of the enterovirus positive samples passed one or both of the coliform standards outlined by the European Economic Community (EEC) bathing water directive (76/160/EEC). Enteroviruses were also detected in 33 of 39 (84.6%) samples tested from 3 inland recreational waters.


Experimental study on subacute neurotoxicity of methotrexate in cats.
An experimental study on the pathogenesis of methotrexate (MTX)-related neurotoxicity including disseminated necrotizing leukoencephalopathy (DNL) was conducted in cats. MTX was administered to the cerebrospinal fluid (CSF) of adult cats using either an intracisternal intermittent instillation (ICI) model or an intraventricular continuous instillation (IVC) model. Furthermore, the synergistic effects of CSF-flow disturbance with kaolin-induced hydrocephalus, and 60Co irradiation were morphologically examined in these models. None of the animals from either the ICI and IVC groups showed DNL, but all animals showed segmental axonal degeneration, suggesting that MTX had a direct toxic effect on the axon. In the ICI groups, no apparent synergistic effect of CSF-flow disturbance and radiation was noted on this axonal change. In the IVC groups, CSF-flow disturbance augmented the degree of the axonal injury. Axonal degeneration and fibrin exudation in the walls of small blood vessels occurred in one animal of the IVC groups with CSF-flow disturbance, suggesting that a toxic effect of MTX on blood vessels is another mechanism of MTX-induced neurotoxicity.


Effect of leg-length discrepancy following total hip arthroplasty on collapse of the contralateral hip in bilateral non-traumatic osteonecrosis of the femoral head.
The purpose of this study was to examine whether leg-length discrepancy (LLD) following unilateral total hip arthroplasty (THA) affects the incidence of contralateral head collapse and subsequent THA in patients with bilateral osteonecrosis, and to determine factors associated with subsequent collapse. We identified 121 patients with bilateral non-traumatic osteonecrosis who underwent THA between 2003 and 2011 to treat a symptomatic hip, and who also exhibited medium-to-large lesions (necrotic area ≥ 30%) in an otherwise asymptomatic non-operated hip. Of the 121 patients, 71 were male (59%) and 50 were female (41%), with a mean age of 51 years (19 to 71) at the time of initial THA. All patients were followed for at least five years and were assessed according to the presence of a LLD (non-LLD vs LLD group), as well as the LLD type (longer non-operated side vs shorter non-operated side group). Overall, 68 hips (56%) became painful and progressed to collapse at a mean of 2.6 years (0.2 to 13.8), resulting in 59 THAs (49%). The five-year collapse-free survival rate for the non-LLD group was 59% (95% confidence interval (CI) 46.8 to 71.8) compared with 45% (95% CI 32.9 to 57.5) for the LLD group (p = 0.036), and 66% (95% CI 55.2 to 77.2) for the longer non-operated side group compared with 32% (95% CI 19.1 to 44.9) for the shorter non-operated side group (p < 0.001). Multivariate regression analyses found that large lesions had a higher risk of collapse than medium-size lesions (odds ratio (OR) 4.19, 95% confidence interval (CI) 1.69 to 10.38; p = 0.002). Meanwhile, patients with a LLD < 3 mm (OR 0.20, 95% CI 0.08 to 0.52; p = 0.001) or a longer non-operated leg (OR 0.11, 95% CI 0.04 to 0.28; p < 0.001) after THA were less likely to experience a subsequent collapse. We found that LLD may be a modifiable risk factor for femoral head collapse. Minimizing LLD and particularly avoiding a shorter non-operated limb after THA may lead to a lower risk of collapse of the asymptomatic hip in patients with bilateral non-traumatic osteonecrosis. Cite this article: Bone Joint J 2019;101-B:303-310.


A second generation linkage map of the human genome based on highly informative microsatellite loci.
The use of simple sequence repeats as polymorphic genetic markers has considerably facilitated the construction of reference maps of the human chromosomes and mapping of Mendelian genetic diseases. These markers display major advantages over other systems and will be widely used for linkage mapping purposes in the forthcoming years.


[The fractionation of chlorine isotopes by the aerobic methylotrophic bacterium Methylobacterium dichloromethanicum grown on dichloromethane].
Methylobacterium dichloromethanicum was found to be able to utilize dichloromethane (DCM) as the source of carbon and energy with the production of biomass, CO2, and HCl. A comparative analysis of abundances of the major DCM isotopomers 35Cl(2)12C1H2, 35Cl37Cl12C1H2 and 37Cl(2)12CH2 made it possible to estimate the fractionation of chlorine isotopes during the bacterial metabolism of DCM. The kinetic chlorine isotope effects for 35Cl37Cl12C1H2 (m/z 86) and 37Cl(2)12C1H2 (m/z 88) relative to 35Cl(2)12C1H2 (m/z 84) turned out to be alpha 86/84 = 1.006 +/- 0.002 and alpha 88/84 = 1.023 +/- 0.003, respectively. The inference is made that the growth of M. dichloromethanicum on DCM is accompanied by the mass-independent fractionation of the DCM isotopomers.


Antihypertensive 1,4-dihydropyridines as correctors of the cystic fibrosis transmembrane conductance regulator channel gating defect caused by cystic fibrosis mutations.
Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis transmembrane conductance regulator (CFTR) Cl- channel gene. CF mutations like deltaF508 cause both a mistrafficking of the protein and a gating defect. Other mutations, like G551D, cause only a gating defect. Our aim was to find chemical compounds able to stimulate the activity of CFTR mutant proteins by screening a library containing approved drugs. Two thousand compounds were tested on Fischer rat thyroid cells coexpressing deltaF508-CFTR and a halide-sensitive yellow fluorescent protein (YFP) after correction of the trafficking defect by low-temperature incubation. The YFP-based screening allowed the identification of the antihypertensive 1,4-dihydropyridines (DHPs) nifedipine, nicardipine, nimodipine, isradipine, nitrendipine, felodipine, and niguldipine as compounds able to activate deltaF508-CFTR. This effect was not derived from the inhibition of voltage-dependent Ca2+ channels, the pharmacological target of antihypertensive DHPs. Indeed, methyl-1,4-dihydro-2,6-dimethyl-3-nitro-4-2(trifluoromethylphenyl)pyridine-5-carboxylate (BayK-8644), a DHP that is effective as an activator of such channels, also stimulated CFTR activity. DHPs were also effective on the G551D-CFTR mutant by inducing a 16- to 45-fold increase of the CFTR Cl- currents. DHP activity was confirmed in airway epithelial cells from patients with CF. DHPs may represent a novel class of therapeutic agents able to correct the defect caused by a set of CF mutations.


Ultra-stable 3D pyridinium salt-based polymeric network nanotrap for selective 99TcO4-/ReO4- capture via hydrophobic and steric engineering.
Selective capture of radioactive 99TcO4- from highly alkaline nuclear waste is highly desirable for environmental remediation and waste disposal. However, the combined features of adsorbents with excellent chemical stability and high capture selectivity for 99TcO4- have not yet been achieved. Herein, we report an ultra-stable 3D pyridinium salt-based polymeric network (TMP-TBPM) nanotrap with remarkable radiation, acid and base stability for selective capture of ReO4- via hydrophobic engineering and steric hindrance, a non-radioactive surrogate of 99TcO4-. The batch capture experiments show that TMP-TBPM has high capture capacity (918.7 mg g-1) and fast sorption kinetics (94.3 % removal in 2 min), which can be attributed to the high density of pyridinium salt-based units on the highly accessible pore channels of 3D interconnected low-density skeleton. In addition, the introduction of abundant alkyl and tetraphenylmethane units into the 3D framework not only greatly enhanced the hydrophobicity and stability of TMP-TBPM, but also significantly improved the affinity toward 99TcO4-/ReO4-, enabling reversible and selective capture of 99TcO4-/ReO4- even under highly alkaline conditions. This study exhibits the great potential of 3D pyridinium salt-based polymeric network nanotrap for 99TcO4-/ReO4- capture from highly alkaline nuclear waste, providing a new strategy to construct high-performance cationic polymeric sorbents for radioactive wastewater treatment.


Sex, Race, and the Quality of Life Factors Most Important to Patients' Well-Being Among Those Seeking Bariatric Surgery.
Evidence suggests obesity-related social stigma and impairment in work function may be the two most detrimental quality of life (QOL) factors to overall well-being among patients seeking weight loss surgery (WLS); whether the relative importance of QOL factors varies across patient sex and race/ethnicity is unclear. We interviewed 574 patients seeking WLS at two centers. We measured patient's health utility (preference-based well-being measure) as determined via standard gamble scenarios assessing patients' willingness to risk death to achieve weight loss or perfect health. Multivariable models assessed associations between patients' utility and five weight-related QOL domains stratified by gender and race: social stigma, self-esteem, physical function, public distress (weight stigma), and work life. Depending on patients' sex and race/ethnicity, mean utilities ranged from 0.85 to 0.91, reflecting an average willingness to assume a 9-15 % risk of death to achieve their most desired health/weight state. After adjustment, African Americans (AAs) reported higher utility than Caucasians (+0.054, p = 0.03), but utilities did not vary significantly by sex. Among Caucasian and AA men, impairment in physical functioning was the most important factor associated with diminished utility; social stigma was also a leading factor for Caucasian men. Among Caucasian women, self-esteem and work function appeared equally important. Social stigma was the leading contributor to utility among AA women; QOL factors did not appear as important among Hispanic patients. AAs reported higher utilities than Caucasian patients. Individual QOL domains that drive diminished well-being varied across race/ethnicity and sex.


Three-month assessment of safety and efficacy of two electric toothbrushes.
This randomised, examiner-blind parallel group study was designed to evaluate the safety and efficacy of a rechargeable oscillating/pulsating toothbrush (Oral-B ProfessionalCare 7000, Oral-B Laboratories; PC 7000) and a battery-operated toothbrush (Crest SpinBrush Pro, Procter & Gamble Company; SBP) in the reduction of gingivitis, bleeding and plaque over a 3-month period. After 12-18 hours of no oral hygiene, subjects had oral tissue examinations, and gingival and plaque assessments to determine eligibility in the study. Subjects were stratified and randomised into treatment groups based on initial whole mouth mean plaque (Turesky modification of Quigley Hein Plaque Index) and gingivitis (Löe & Silness Gingival Index) scores and gender. Subjects were instructed to brush twice daily with their assigned toothpaste and toothbrush. Clinical parameters were assessed at baseline, and after 1 and 3 months of use. Within treatment comparisons from baseline were analysed using t-test; between treatment comparisons were analysed using ANOVA. Data were obtained from 92 subjects (PC 7000 n=45; SBP n=47). No significant differences were found in baseline plaque, gingivitis and bleeding between groups. Both treatment groups had significant reductions from baseline in plaque, gingivitis and bleeding scores. PC 7000 demonstrated significantly greater reductions compared to SBP in whole mouth plaque at 1 month: 0.39 +/- 0.43 vs. 0.16 +/- 0.42 and 3 months: 0.32 +/- 0.48 vs. 0.04 +/- 0.41. PC 7000 also demonstrated significant reductions compared to SBP in gingivitis at 3 months for whole mouth: 0.14 +/- 0.09 vs. 0.10 +/- 0.10 and approximal areas: 0.11 +/- 0.08 vs. 0.08 +/- 0.09. There were no significant differences between toothbrushes in bleeding at either time point: Safety examinations revealed no apparent difference in soft and hard tissue abnormalities between groups. The PC 7000 toothbrush demonstrated significantly greater reductions in plaque and gingivitis compared to the SPB over a 3-month period.


Utility of the Sindbis replicon system as an Env-targeted HIV vaccine.
Sindbis replicon-based vaccine vectors are designed to combine the immunostimulatory properties of replicating viruses with the superior safety profile of non-replicating systems. In this study we performed a detailed assessment of Sindbis (SIN) replicon vectors expressing HIV-1 envelope protein (Env) for the induction of cell-mediated and humoral immune responses in a small animal model. SIN-derived virus-like particles (VLP) elicited Env-specific antibody responses that were detectable after boosting with recombinant Env protein. This priming effect could be mediated by replicon activity alone but may be enhanced by Env attached to the surface of VLP, offering a potential advantage for this mode of replicon delivery for Env based vaccination strategies. In contrast, the Env-specific CTL responses that were elicited by SIN-VLP were entirely dependent on replicon activity. SIN-VLP priming induced more durable humoral responses than immunization with protein only. This is important from a vaccine perspective, given the intrinsic tendency of Env to induce short-lived antibody responses in the context of vaccination or infection. These results indicate that further efforts to enhance the magnitude and durability of the HIV-1 Env-specific immune responses generated by Sindbis vectors, either alone or as part of prime-boost regimens, are justified.


Cold sensitivity after replantation in relation to arterial circulation and vasoregulation.
Cold intolerance is a common problem after injuries to the hand. To elucidate the mechanism of this cold sensitivity, eight patients with successful replantations of amputations of the thumb or index finger were studied one and half years later. In six patients the Doppler signal from the anastomosed arteries was normal and in two patients signs of mild stenosis were found. All patients had normal or slightly reduced basal finger systolic pressure in the replanted finger. Six had signs of marked vasospasm. Alpha-receptor blockade had no effect on the cold-induced decrease in finger systolic pressure in the replanted finger. It is concluded that cold intolerance after hand injuries results from a defect in vasoregulation and is not caused by organic insufficiency of the circulation.


A phase I evaluation of the effect of curcumin on dose-limiting toxicity and pharmacokinetics of irinotecan in participants with solid tumors.
Curcumin inhibits UDP-glucuronyltransferases, a primary metabolic pathway for cancer chemotherapeutic agents like irinotecan. Concurrent administration of both agents may exacerbate irinotecan toxicity. We conducted this phase I study to determine the safety of concurrent curcumin and irinotecan administration. Ten participants with advanced solid tumors received one of four doses (1, 2, 3, and 4 g) of a curcumin phosphatidylcholine complex (PC) orally daily, and 200 mg/m2 of i.v. infusion irinotecan on days 1 and 15 of a 28-day cycle, to determine the maximum tolerated dose (MTD) of PC. Thirteen participants received 4 g of PC (MTD) to assess the effect on the pharmacokinetic (PK) properties of irinotecan and its metabolites, SN-38 and SN-38G. Irinotecan, SN-38, and SN-38G exposure equivalence with and without curcumin was assessed using area under the plasma concentration-time curves from 0 to 6 h (AUC0-6h ). Safety assessments and disease responses were also evaluated. The combination of irinotecan and PC was well-tolerated. Because there was no dose limiting toxicity, the maximum dose administered (4 g) was defined as the recommended phase II dose of PC. PC did not significantly alter the plasma exposure and other PK properties of irinotecan and its metabolites. There was no apparent increase in the incidence of irinotecan-associated toxicities. The objective response rate was 3/19 (22%, 95% confidence interval [CI]: 5-39%), median progression free survival and overall survival (n = 23) were 4 months (95% CI: 2.9-8.9 months) and 8.4 months (95% CI: 3.7 - not evaluable [NE]), respectively. Future studies are required to evaluate the efficacy of this combination.


PCR amplification and DNA sequencing of SRV-2 from archived tumor tissues.
Fibroproliferative tumors, known as retroperitoneal fibromatosis (RF) are predominantly associated with type D simian retrovirus serotype 2 (SRV-2) infections in pigtailed macaques (Macaca nemestrina) at the University of Washington Regional Primate Research Center (UWRPRC). In this report we examined genetic changes that occurred in SRV-2/Washington (SRV-2/WA) from archived RF tissues and from fresh tissue and lymphocytes. DNA was extracted from paraffin-embedded RF tumor tissues and was used to amplify a portion of the SRV-2/WA genome encoding an open reading frame, 3'orf. Since 3'orf is encoded in an alternate reading frame within the outer membrane glycoprotein (gp70) env gene, sequences of 3'orf also provide information on a portion of env gp70. To define the changes that have occurred in this portion of the viral genome, we compared the nucleotide and deduced amino acid sequences amplified from the various tumor tissues. The paraffin section PCR revealed that all samples containing amplifiable DNA harbored SRV-2/WA genomes. DNA sequence analysis showed that the 3'orf/gp70 portion of the SRV-2/WA genome was nearly identical in all tissues but was distinguishable from the prototype SRV-2/OR isolate from Celebes black macaques (M. nigra). A T-cell activating domain (Env residues 233-249) was 100% conserved in the SRV-2/WA isolates.


Sex differences among patients receiving ticagrelor monotherapy or aspirin after coronary bypass surgery: A prespecified subgroup analysis of the TiCAB trial.
There is limited evidence on the association of sex with outcomes among patients undergoing coronary bypass surgery (CABG) and treated with ticagrelor monotherapy or aspirin. This was a pre-specified sub-analysis of TiCAB, an investigator-initiated placebo-controlled randomized trial. Primary efficacy endpoint was the composite of cardiovascular death, myocardial infarction, stroke, or repeat revascularization 1 year after CABG. Safety endpoint was BARC type 2, 3 or 5 bleeding. A total of 280 (15.0%) women and 1579 (85.0%) men were included. Compared with men, women were older (66.1 ± 10.2 vs. 70.1 ± 9.3 years) with more acute presentation (17.0% vs 21.1%). The incidence of the primary endpoint was similar between women and men (9.2% vs. 8.9%, HR 1.08, 95%CI 0.71-1.66, P = 0.71). Cardiovascular death occurred more often in women (2.9% vs 1.0%, adjusted HR 2.87, 95%CI 1.23-6.70, P = 0.02). The incidence of bleeding was similar between the sexes (2.2% vs. 2.5%, HR 0.91, 95% CI 0.51-1.65, P = 0.77). Ticagrelor vs aspirin was associated with a similar risk of the primary endpoint in women (10.6% vs. 7.9%, HR 1.39, 95%CI 0.63-3.05, P = 0.42) and men (9.5% vs. 8.2%, HR 1.15, 95%CI 0.82-1.62, P = 0.41;pinteraction = 0.69), and a similar risk of bleeding in women (2.9% vs. 1.4%, HR 2.09, 95%CI 0.38-11.41, P = 0.40) and men (2.2% vs. 2.8%, HR 0.80, 95%CI 0.42-1.52, P = 0.49;pinteraction = 0.35). Among women and men undergoing CABG, ticagrelor monotherapy was associated with a similar risk of the primary efficacy endpoint and bleeding compared with aspirin. The risk of cardiovascular death was increased in women irrespective of antiplatelet therapy.


[Case of early gastric carcinoma (type IIc) and macroscopic submucosal tumors containing ectopic glands].
A 69 year-old male was admitted to our hospital with early gastric carcinoma (Type IIc) and two macroscopic submucosal tumors. He had suffered from gastroduodenal ulcer for about forty years; postoperative histologic examination revealed the gastric ectopic glands to be a paracancerous lesion.


Threshold for retinal damage associated with the use of high-power neodymium-YAG lasers in the vitreous.
Ultrashort, Q-switched or mode-locked, neodymium-YAG laser pulses focused within 2 mm of the retina caused reproducible retinal damage in four eyes of two monkeys and in four eyes of three rabbits. The distance of the laser focus from the retina for clinically observed threshold retinal damage was characterized for pulse energies up to 9 mJ. For the 2- to 6-mJ pulse energies necessary to rupture vitreal membranes in clear media in rabbits, the high-power laser pulses could not be focused within 2 mm of the retina without substantial risk of damaging the underlying retina. These laser pulses did not rupture vitreal membranes in hazy ocular media that prevented precise focusing. The retinal damage was somewhat greater than expected for retinal absorption of 1.06-micron laser energy, suggesting that secondary effects such as self-focusing and shock waves emanating from the focus may be important.


Dose-dependent effects of isoflurane on regional activity and neural network function: A resting-state fMRI study of 14 rhesus monkeys: An observational study.
The dose-dependent effect of isoflurane on cerebral regional activity and functional connectivity (FC) in 14 rhesus monkeys was investigated using resting-state functional MRI. Amplitude of low-frequency fluctuations (ALFF) decreased in the cerebellum, visual cortex, and cortico-subcortical network when the isoflurane dose changed from 1.0 to 1.3 MAC. ALFF decreased in the arousal system, cerebellum, sensory, visual areas, cortico-subcortical network and default mode network and increased in the bilateral dorsal prefrontal cortices, frontal eye fields and motor-related areas from 1.0 to 1.6 MAC. FC of the default mode network, frontal-parietal, cortico-subcortical, motor, sensory, auditory and visual areas was reduced when isoflurane increased from 1.0 to 1.3 MAC. FC decreased in more widespread areas, especially in regions of cortico-subcortical networks and limbic systems, when isoflurane further increased from 1.0 to 1.6 MAC. Both dose-dependent decreased and increased ALFF were separately observed, while FC deteriorated as the anesthesia deepened. These results suggest that changes continue to occur past the loss of consciousness, and the dose-dependent effects of isoflurane are different with regard to regional function and neural network integration.


Synergistic effect of bismuth subgallate and borneol, the major components of Sulbogin, on the healing of skin wound.
Most skin lesions heal delay and even heal efficiently within 1-2 weeks, the healed tissue is neither aesthetically nor functionally perfect. Therefore, facilitating skin healing rate and controlling healed skin quality are major aims of drug treatment for a wound event. Bismuth subgallate (BS) and Borneol (BO) are the two components of Sulbogin, a new Vaseline-based wound healing ointment, one for treating skin wounds. Although BO has antibiotic function, while BS is widely used clinically, neither has been used specifically for wound healing. The experiment described here aimed to study the effect of BS and BO on the healing of skin wounds. This study also compared the effects of BS and BO with Flamazine cream, which is currently the most popular drug for wound healing in hospitals. Full-thickness wounds (3 cm x 3 cm x 0.2 cm) were created on the back of adult male Sprague-Dawley rats. BS, BO, BS+BO, and Flamazine were then evenly applied to cheesecloth and placed over the lesion areas. The drug patches were replaced every 2-3 days until the wound areas were completely covered by epidermis in any kinds of drug treatment. The combined BS and BO treatment had the best effect on healing by decreasing lesion area, while increasing granulation tissue formation, re-epithelialization, eating behavior and reconstitution of skin appendages. This investigation showed that BS and BO have a synergistic effect on the skin wound restoration.


Indoor endotoxin, proximity to a major roadway, and severe asthma exacerbations among children in Puerto Rico.
Few studies have examined concurrent exposure to household endotoxin and traffic-related air pollution in relation to childhood asthma, yet both factors are associated with asthma outcomes. To examine whether proximity to a major roadway (a traffic-related air pollution proxy) modifies the estimated effects of indoor endotoxin on asthma outcomes in children. Cross-sectional study of 200 children with asthma (ages, 6-14 years) living in Puerto Rico. Residential distance to a major roadway was calculated as the distance from the participant's residential US census block centroid to the nearest major road. The outcomes of interest were severe asthma exacerbations, missed school days for asthma, atopy, lung function, and bronchodilator response (BDR). Logistic, linear, or negative binomial regression was used for the multivariable analysis. In the multivariable analysis, there was an interaction between indoor endotoxin and residential distance to a roadway on severe asthma exacerbations (P = .02) and BDR (P = .07). In an analysis stratified by distance to a roadway, each log10-unit increase in endotoxin was associated with 4.21 times increased odds of severe asthma exacerbations among children living within 499 m (the lower 3 quartiles of residential distance) to a road (95% confidence interval, 1.5-12.0). Among subjects living further than 499 m away from a roadway, each log10-unit increase in endotoxin was associated with reduced odds of severe asthma exacerbations (odds ratio, 0.03; 95% confidence interval, 0.001-0.67). Similar but less substantial findings were observed for BDR. Our findings suggest that residential proximity to a major road modifies the estimated effect of endotoxin on severe asthma exacerbations in children.


Emergency percutaneous tracheostomy in trauma patients: an early experience.
In recent years, percutaneous tracheostomy (PCT) has become a routine practice in many hospitals. In the early publications, most authors considered adverse conditions such as short or fat neck or obesity as relative contraindications, whereas cervical injury and emergency were regarded as absolute contraindications. More recently, several reports demonstrated the safety and feasibility of PCT in patients with some of the above contraindications. We, like many others, gradually reduced the contraindications and expanded the indications for PCT. In this paper, we report our early experience with emergency PCT in trauma patients. Ten adult patients suffering from multiple injuries after motor vehicle accident (7) or severe head and neck burns (3) required emergency surgical airway control after failure to accomplish orotracheal intubation. A modified Griggs' technique was used by experienced thoracic surgeons. Recorded data included patient demographics, clinical and anatomic conditions, length of procedure, and complications. Short-term follow-up was performed in the hospital by thoracic staff surgeons. Long-term follow-up was carried out in the outpatient clinic. Six male and 4 female patients underwent emergency PCT. The mean time from skin incision to intubation was 5.5 minutes including the oxygen insufflation period. There was no failure, no procedure-related complication, and no conversion to open technique. Five patients survived and underwent uneventful decannulation. In approximately 1 year of follow-up, there were no clinical symptoms or signs of complications related to the tracheostomy. Emergency PCT using a modified Griggs' technique is feasible and safe. In experienced hands, it might be even easier and faster than the open surgical tracheostomy.


PI16 is expressed by a subset of human memory Treg with enhanced migration to CCL17 and CCL20.
The peptidase inhibitor PI16 was shown previously by microarray analysis to be over-expressed by CD4-positive/CD25-positive Treg compared with CD4-positive/CD25-negative Th cells. Using a monoclonal antibody to the human PI16 protein, we found that PI16-positive Treg have a memory (CD45RO-positive) phenotype and express higher levels of FOXP3 than PI16-negative Treg. PI16-positive Treg are functional in suppressor assays in vitro with potency similar to PI16-negative Treg. Further phenotyping of the PI16-positive Treg revealed that the chemokine receptors CCR4 and CCR6 are expressed by more of the PI16-positive/CD45RO-positive Treg compared with PI16-negative/CD45RO-positive Treg or Th cells. PI16-positive Treg showed enhanced in vitro migration towards the inflammatory chemokines CCL17 and CCL20, suggesting they can migrate to sites of inflammation. We conclude that PI16 identifies a novel distinct subset of functional memory Treg which can migrate to sites of inflammation and regulate the pro-inflammatory response at those sites.


Surgical resection of a thymoma developed in a case with isolated persistent left superior vena cava.
Persistent left superior vena cava (PLSVC) is one of the most common vascular abnormalities in the chest. In approximately 10 % of cases, the right superior vena cava is missing, which is called isolated persistent left superior vena cava (IPLSVC). The case is an 85 years-old female. An anterior mediastinal tumor was accidentally revealed when the patient was admitted after a traffic accident. As the tumor became larger within four months, a thymectomy was planned. The anterior mediastinal tumor was in front of the ascending aorta, which was close to the confluence of the left and right brachiocephalic veins in normal anatomy. However, in this case, the right superior vena cava was missing, and the right brachiocephalic vein flowed into the left superior vena cava by the chest computed tomography. Preoperative examinations found no accompanying cardiac abnormality. Robot-assisted thymectomy was performed. No tumor infiltration was observed in the right brachiocephalic vein. No abnormality was found in either phrenic nerve. The tumor could be safely resected, and her postoperative course was uneventful. The pathological diagnosis was a thymoma. A case of thymectomy with IPLSVC is quite rare. A careful observation of the preoperative computed tomography images helps to diagnose IPLSVC. Technically, thymectomy was not much different from normal, other than the reversed location of the veins. However, it should be noted that IPLSVC cases may have cardiac malformations. Thymectomy for thymoma with IPLSVC can be safely performed when the left and right veins are reversed.


Examination of the laryngeal musculature and recurrent laryngeal nerves of zebra (Equus burchelli) for evidence of idiopathic laryngeal hemiplegia.
This study was undertaken in order to determine whether wild Burchell's Zebra (Equus burchelli) are affected by Idiopathic Laryngeal Hemiplegia (ILH). ILH has a high prevalence in the domestic horse and a negative control population has not been identified. The work was conducted in two phases on two large farms in central Kenya during routine culls. The larynges of 42 zebra were removed and the left and right intrinsic laryngeal adductor and abductor muscles and recurrent laryngeal nerves were isolated. The muscles were weighed and biopsy samples of left and right recurrent laryngeal nerve and cricoarytenoideus lateralis muscles from 16 animals were taken for histological examination. Left adductor muscles were observed to be significantly lighter than right adductor muscles (p = 0.001). There was no significant difference between the weights of left and right laryngeal abductors (p = 0.260). There was no light microscopic evidence of pathological changes consistent with ILH in any of the muscle or nerve samples. Zebra do not appear to suffer from ILH and equids unaffected by ILH can have significantly lighter muscles on the left of the larynx. Mild asymmetries of the larynx on palpation may not be a reliable indicator of ILH. Zebra may be a potentially useful negative control animal for observational ILH studies.


The metabolism of dibenzylamine: identification of NN-dibenzylhydroxylamine as the major in vitro metabolic product from rabbit fortified hepatic homogenates.
A study of the in vitro metabolism of dibenzylamine with fortified rabbit liver 9000 g supernatant fractions has shown that N-oxidation is the major metabolic process. Of the total amount of dibenzylamine metabolized, 90% is converted to NN-dibenzylhydroxylamine. The primary amine, benzylamine, is formed to the extent of only 6% of total substrate metabolized. The identity of the major metabolite, NN-dibenzylhydroxylamine has been established by thin-layer chromatography, gas-liquid chromatography and combined gas-liquid chromatography-mass spectrometry.


Transthoracic defibrillation threshold of sine and trapezoidal waveforms in defibrillation.
The transthoracic defibrillation threshold of trapezoidal (2.5-, 5-, 10-, 15-, 20-, 40-, and 55-msec duration) and damped sine waveforms were established in groups of anesthetized greyhounds. Of the seven trapezoidal waveforms, the threshold current was lowest for the 20-msec pulse but was not significantly different for the 5-20-msec pulses (p greater than 0.05). The threshold delivered energy was not significantly different for the 2.5-20-msec waveforms. The 40- and 55-msec pulses had significantly greater delivered energies than the shorter pulses (p less than 0.01). The transthoracic defibrillation threshold of two trapezoidal (5- and 20-msec) and three damped sine waves (Lown, Edmark, and Belfast) showed that the threshold transthoracic current and energy were not significantly different for the 5-msec trapezoid and three sine waves (p greater than 0.05). The threshold transthoracic energy for the 20-msec trapezoid was significantly greater than that of the four other waveforms. The threshold current for the 20-msec trapezoid was significantly less than that of the Belfast and Edmark waveforms only. Thus, none of the three damped sine waveforms was superior, with the 5-msec trapezoid producing similar results. Although the 20-msec trapezoid had the lowest threshold current, it had the highest delivery energy.


Good interobserver agreement was attainable on outcome adjudication in patients with stable coronary heart disease.
In clinical trials, agreement on outcomes is of utmost importance for valid estimation of intervention effects. As there is limited knowledge about adjudicator agreement in cardiology, we examined the level of agreement among three cardiology specialists adjudicating all possible events in a randomized controlled clinical trial of patients with stable coronary heart disease. All information (hospital records, death certificates, etc.) was forwarded to two randomly selected blinded adjudicators. If they disagreed, the third arbiter had to choose the more likely of the two alternatives. Files of 5,475 nonfatal and 362 fatal events were evaluated. For nonfatal outcomes, pairwise kappa values ranged from 0.75 to 0.80. The three adjudicators had 4.3%, 9.5%, and 6.1% of their nonfatal outcome classifications overruled by their arbiter. If stable angina pectoris, unstable angina pectoris, and acute myocardial infarction were treated as one, agreement increased minimally. For fatal outcomes, the pairwise kappa values ranged from 0.65 to 0.90. The three adjudicators had 12%, 9%, and 10% of their death classifications overruled. Specialists in cardiology can attain a reasonably high agreement on outcomes in patients with stable coronary heart disease.


Synergistic cell inactivation by cis-dichlorodiammineplatinum in combination with 1-propargyl-5-chloropyrimidin-2-one.
A synergistic effect with respect to inactivation of human NHIK 3025 cells cultured in vitro was displayed when treatment with cis-dichlorodiammineplatinum(II) (cis-DDP) and the mitotic inhibitor 1-propargyl-5-chloropyrimidin-2-one (NY 3170) were given in simultaneous combination. Cell inactivation was measured by loss of colony-forming ability. Treatment with NY 3170 alone produced no significant inactivation at concentrations up to 2 mM. However, treatment with NY 3170 in combination with cis-DDP induced increased cell inactivating effects equal to a doubling of either the concentration of cis-DDP or treatment time. Scheduling of NY 3170 treatment in relationship to a 1 h cis-DDP pulse revealed that synergism occurred only when the two drugs were present simultaneously. The inactivating effect of 10 microM cis-DDP in combination with 2 mM NY 3170 given to synchronized NHIK 3025 cells at various stages of the cell cycle was also determined. For cells treated in S or in G2 + M cell survival was reduced by a factor of 5 after a 1 h treatment with the drug combination as compared to similar treatment with cis-DDP alone. The cells appeared to be most sensitive at the time of initiation of DNA synthesis. Here cell survival was reduced by a factor of 100 following treatment with the drug combination than following treatment with cis-DDP alone. Measurement of cell-associated platinum by atomic absorption spectroscopy indicated that cellular uptake of cis-DDP was increased when NY 3170 was simultaneously present during drug treatment.


Endoscopic Ultrasound-Guided Gastro-gastrostomy Using Lumen Apposing Metal Stent for Management of High-Grade Gastric Outlet Obstruction Secondary to Vertical Banded Gastroplasty.
Vertical banded gastroplasty (VBG) is associated with an increased risk of pouch stricture secondary to narrowing of the banded gastroplasty outlet. We describe a case of a 60-year-old male with stage 4 small cell lung cancer presenting with recurrent gastric outlet obstruction secondary to stenosis at the site of VBG. Given a prior history of failed standard endoscopic maneuvers including balloon dilation and steroid injection, the decision was made to create an endoscopic ultrasound-guided gastro-gastrostomy using lumen opposing metal stent. The patient did well post procedure and was able to tolerate a soft mechanical diet within 24 h. He was subsequently discharged to hospice for his advanced metastatic disease.


Computational modeling of the direct hydride transfer mechanism for the MAO catalyzed oxidation of phenethylamine and benzylamine: ONIOM (QM/QM) calculations.
Monoamine oxidases are two isozymic flavoenzymes which are the important targets for drugs used in the treatment of depression, Parkinson and Alzheimer's diseases. The catalytic reaction taking place between the cofactor FAD and amine substrate is still not completely understood. Herein we employed quantum chemical methods on the recently proposed direct hydride transfer mechanism including full active site residues of MAO isoforms in the calculations. Activation free energy barriers of direct hydride transfer mechanism for MAO-A and MAO-B were calculated by ONIOM (our own n-layered integrated molecular orbital + molecular mechanics) method with QM/QM (quantum mechanics:quantum mechanics) approach employing several density functional theory functionals, B3LYP, WB97XD, CAM-B3LYP and M06-2X, for the high layer. The formation of very recently proposed αC-flavin N5 adduct inside the enzyme has been investigated. ONIOM (M06-2X/6-31+G(d,p):PM6) results revealed that such an adduct may form only in MAO-B suggesting slightly different hydride transfer mechanisms for MAO-A and MAO-B.


Models of multiple system atrophy.
Multiple system atrophy (MSA) is a neurodegenerative disease with diverse clinical manifestations, including parkinsonism, cerebellar syndrome, and autonomic failure. Pathologically, MSA is characterized by glial cytoplasmic inclusions in oligodendrocytes, which contain fibrillary forms of α-synuclein. MSA is categorized as one of the α-synucleinopathy, and α-synuclein aggregation is thought to be the culprit of the disease pathogenesis. Studies on MSA pathogenesis are scarce relative to studies on the pathogenesis of other synucleinopathies, such as Parkinson's disease and dementia with Lewy bodies. However, recent developments in cellular and animal models of MSA, especially α-synuclein transgenic models, have driven advancements in research on this disease. Here, we review the currently available models of MSA, which include toxicant-induced animal models, α-synuclein-overexpressing cellular models, and mouse models that express α-synuclein specifically in oligodendrocytes through cell type-specific promoters. We will also discuss the results of studies in recently developed transmission mouse models, into which MSA brain extracts were intracerebrally injected. By reviewing the findings obtained from these model systems, we will discuss what we have learned about the disease and describe the strengths and limitations of the models, thereby ultimately providing direction for the design of better models and future research.


Tibioperoneal Short Circuiting for Stump Neuroma Pain in Amputees: Revival of an Old Technique.
Stump neuroma pain in amputees can be quite challenging. Surgical treatment may be largely subdivided into neuromodulative and non-neuromodulative methods. The latter includes neurocapsis, insertion of nerve stump into the bone marrow, centro-central short circuit (CCSC), and coverage with vascularized soft tissue flaps. CCSC was shown to be extremely effective in alleviation of pain. Reports on CCSC for the treatment of stump neuroma pain have disappeared from the literature, with a shift toward neuromodulation for the treatment of pain irrespective of etiology. We observed 8 lower limb amputees undergoing CCSC of the sciatic nerve during a follow-up of 12 years. All had the same stump neuroma pain rendering them unable to wear their prostheses. The sciatic nerve was explored at the midthigh area, much proximal to the amputation site, and a short circuit was established between the tibial and peroneal parts of the nerve. Assessment was by means of pain quantification as per the quadruple visual analogue scale, medication intake, and ability to use prostheses. The median worst quadruple visual analogue scale before surgery was 8.0. After surgery it decreased to 2.5 (P = 0.00094). Medication intake was reduced from regular intake of a combination of opioids, nonsteroidal anti-inflammatory drugs, tricyclic antidepressants, and pregabalin in all patients to irregular intake of nonsteroidal anti-inflammatory drug alone in 3 of 8 patients. All patients were able to wear their limb prosthesis since surgery. CCSC is a simple, effective, and long-lasting method to treat painful stump neuromas in amputees. It should be strongly considered in deserving cases before resorting to neuromodulative methods.


Multilevel Factors Shaping Awareness of and Attitudes Toward Pre-exposure Prophylaxis for HIV Prevention among Criminal Justice-Involved Women.
Although pre-exposure prophylaxis (PrEP) is a key tool in HIV prevention efforts, little is known about PrEP as a prevention strategy for criminal justice-involved (CJI) women. The purpose of this study was to examine multilevel factors shaping PrEP awareness and acceptability among CJI women. Between January 2017 and December 2017, we conducted 52 interviews with CJI women at high risk for HIV and stakeholders from the criminal justice (CJ) and public health (PH) systems. Interviews explored awareness of PrEP and the multilevel factors shaping PrEP acceptability. Data were analyzed using inductive thematic analysis and executive summaries. Atlas.ti facilitated analyses. The majority of CJI women (n = 27) were, on average, 41.3 years, from racial and ethnic minority groups (56% Black/African-American; 19% Latinx) and reported engaging in recent high-risk behavior (nearly 60% engaged in transactional sex, 22% reported ≥ 4 sexual partners, and 37% reported injection drug use). Of system stakeholders (n = 25), 52% represented the CJ sector. Although CJI women were generally unaware of PrEP, attitudes toward PrEP were enthusiastic. Barriers to PrEP acceptability included medication side effects (individual level); distrust in HIV prevention mechanisms (community level); lack of local HIV prevention efforts among high-risk women (public policy/HIV epidemic stage level). Factors promoting PrEP included perceived HIV risk (individual level); PrEP being an HIV prevention method that women can control without partner negotiation (social and sexual network level); and availability of public health insurance (community level). Despite low awareness of PrEP, CJI women expressed positive attitudes toward PrEP. To improve PrEP access for CJI women, implementation efforts should address barriers and leverage facilitators across multiple levels to be maximally effective.


Elevated oxidized glutathione in cystinotic proximal tubular epithelial cells.
Cystinosis, the most frequent cause of inborn Fanconi syndrome, is characterized by the lysosomal cystine accumulation, caused by mutations in the CTNS gene. To elucidate the pathogenesis of cystinosis, we cultured proximal tubular cells from urine of cystinotic patients (n = 9) and healthy controls (n = 9), followed by immortalization with human papilloma virus (HPV E6/E7). Obtained cell lines displayed basolateral polarization, alkaline phosphatase activity, and presence of aminopeptidase N (CD-13) and megalin, confirming their proximal tubular origin. Cystinotic cell lines exhibited elevated cystine levels (0.86 +/- 0.95 nmol/mg versus 0.09 +/- 0.01 nmol/mg protein in controls, p = 0.03). Oxidized glutathione was elevated in cystinotic cells (1.16 +/- 0.83 nmol/mg versus 0.29 +/- 0.18 nmol/mg protein, p = 0.04), while total glutathione, free cysteine, and ATP contents were normal in these cells. In conclusion, elevated oxidized glutathione in cystinotic proximal tubular epithelial cell lines suggests increased oxidative stress, which may contribute to tubular dysfunction in cystinosis.


Local structure of human hair spatially resolved by sub-micron X-ray beam.
Human hair has three main regions, the medulla, the cortex, and the cuticle. An existing model for the cortex suggests that the α-keratin- based intermediate filaments (IFs) align with the hair's axis, but are orientationally disordered in-plane. We found that there is a new region in the cortex near the cuticle's boundary in which the IFs are aligned with the hair's axis, but additionally, they are orientationally ordered in-plane due to the presence of the cuticle/hair boundary. Further into the cortex, the IF arrangement becomes disordered, eventually losing all in-plane orientation. We also find that in the cuticle, a key diffraction feature is absent, indicating the presence of the β-keratin rather than that of the α-keratin phase. This is direct structural evidence that the cuticle contains β-keratin sheets. This work highlights the importance of using a sub-micron x-ray beam to unravel the structures of poorly ordered, multi-phase systems.


[Studies on chemical constituents from roots of Mirabilis jalapa].
To investigate the anti-HIV constituents from the root of Mirabilis jalapa. The compounds were isolated by column chromatography on silica gel, Sephadex LH - 20, MCI-gel CHP-20P and RP-18. The structure were identified by means of NMR and MS analyses (1H-NMR, 13C-NMR, MS). Eleven compounds were isolated and identified as astragaloside II (1), astragaloside II (2), astragaloside IV (3), astragaloside VI (4), flazin (5), 4'-hydroxy-2, 3-dihydroflavone 7-beta-D-glucopyranoside (6), gingerglycolipid A (7), 3, 4-dihydroxybenzaldehyd (8), p-hydroxybenzaldehyde (9), beta-sitosterol (10) and daucosterol (11). Compounds 1-9 were obtained from this genus for the first time.


Retinoid receptor alpha and Beta expression in serous ovarian tumors.
The expression of retinoid acid receptors alpha (RARalpha) and beta (RARbeta) and estrogen receptor alpha (ERalpha) was assessed by immunohistochemistry and Western blotting in normal ovaries, serous cystadenoma (n = 20), serous borderline (n = 14), and serous ovarian cancer (n = 47) and was correlated in cancer cases with stage, grade, progress-free survival (PFS), and survival. RARalpha was increasingly expressed in benign cystadenomas, borderline, and low-stage and advanced-stage neoplasms (p < 0.001). In stage III, G3 serous carcinoma, increased RARalpha expression was an independent prognostic factor associated with lower chemoresponse to first-line chemotherapy (taxol and carboplatin) and shorter PFS (p < 0.002).RARbeta and ERalpha expression did not correlate with RARalpha tumor characteristics or PFS and survival.


Characterization of physico-chemical properties and antioxidant activity of oil from seed, leaf and stem of purslane (Portulaca oleracea L.).
Purslane (Portulaca oleracea L.) is a member of the family of portulacaceae and is a very good source of alpha-linoleic acid. Alpha-linoleic is an omega-3 fatty acid which plays an important role in human growth, development and in preventing diseases. Characterization of the physicochemical properties and antioxidant activity of oil from seeds, leaves and stems of purslane, and appraisal of its viability is the main purpose of this study. In this work, physicochemical properties of oil from seed, leaf and stem of purslane were characterized and the highest oil content was observed in seed (11.25 %) using solvent extraction method. The highest specific gravity was recorded for seed oil (1.10 w/w) followed by leaf oil (0.73 w/w) with non-significant difference. Moreover, oil extracted from purslane stem was found to be the least in oil content (2 %) and specific gravity (0.14 w/w); but the highest in acid value (13.32 w/v) and percentage free fatty acid. The peroxide value was found to be the highest for seed oil and the least for stem with significant difference. The highest ascorbic acid content was recorded for purslane seed oil (41.67 %) followed by leaf oil (32.29 %). Likewise, the highest 1, 1- diphenyl-2-picrylhydrazine activity was obtained for leaf oil (12.55 %) followed by seed oil (2.05 %). The plot for PC2 vs PC1 showed the oil content, specific gravity, peroxide value, ascorbic acid and 1, 1- diphenyl-2-picrylhydrazine having close PC1 and PC2 scores with vector angle < 90 0 showing correlated effects. Characterizing physical and chemical properties and antioxidant activity of oil from seeds, leaves and stems of purslane is crucial in providing necessary information for the utilization of the oil content.


Role of cyclin E and p53 expression in progression of early gastric cancer.
BACKGROUND: To elucidate the role that cyclin E overexpression plays in the progression of early gastric cancer, we examined the expression of cyclin E and p53, as abnormal p53 expression is linked with cyclin E overexpression in exerting adverse affects on the cell cycle.METHODS: Specimens from 108 early gastric cancers were stained by an immunohistochemical method, using anti-cyclin E and anti- p53 antibodies.RESULTS: The positivity rate of cyclin E expression in early gastric cancer was 33% (36/108). Cyclin E-positive tumors invaded more deeply ( P < 0.05), infiltrated lymphatic vessels more frequently ( P < 0.01), showed a higher incidence of differentiated cancer ( P < 0.01), and more often expressed p53 ( P < 0.01) than cyclin E-negative tumors. Differentiated cancers showing coexpression of cyclin E and p53 were more likely to metastasize to the lymph nodes.CONCLUSIONS: Overexpression of cyclin E may promote the progression of early gastric cancer.


The Biochemical and Pathological Correlates of Successful Semen Cryopreservation From Patients With Testicular Cancer: A Single Tertiary Center Experience.
To determine the predictors for success with regard to semen cryopreservation and good semen quality of patients with testicular cancer. A total of 50 men (aged 16-36 years) with testicular cancer, referred for semen cryopreservation prior to gonadotoxic treatment, were included. Age, alpha fetal protein (α-FP), beta human chorionic gonadotropin, lactate dehydrogenase, clinical staging, tumor volume, and pathological reports were evaluated as correlates of successful semen cryopreservation and good semen quality. The overall success rate was 52%. α-FP (4113.1 ng/mL vs 81.2 ng/mL) and tumor volume (77.7 mL vs 25.5 mL) were significantly higher in the failure group as compared to the success group. The seminoma to nonseminomatous germ cell tumor ratio was lower in the failure group as compared to the success group (9/17 vs 3/21). There was nearly a significant difference (P = .066). The optimal cutoff value for α-FP > 1000 ng/mL showed the highest Youden index (0.689) and resulted in a sensitivity of 0.625 and specificity of 1.0 for predicting poor outcome. In terms of multivariate analysis, the α-FP (P = .013), tumor volume (P = .047), and α-FP > 1000 ng/mL (P = .010) were significantly associated with poor semen quality and failure to preserve semen. Sperm quality was found to be higher in the seminoma versus the nonseminomatous germ cell tumor patients in: sperm concentration (21.5 million/mL vs 11.8 million/mL, P < .027). Furthermore, tumor volume is correlated to α-FP (P = .018) and is weakly correlated to lactate dehydrogenase (P = .067) CONCLUSION: Elevated α-FP and tumor volume are independently poor factors for semen quality and semen cryopreservation. In clinical use, α-FP is a noninvasive tool to predict the success of semen cryopreservation and patients with α-FP > 1000 ng/mL should be informed of the higher risk of poor semen quality and semen cryopreservation concerns.


Regulated nuclear accumulation of the yeast hsp70 Ssa4p in ethanol-stressed cells is mediated by the N-terminal domain, requires the nuclear carrier Nmd5p and protein kinase C.
Cytoplasmic proteins of the hsp70/hsc70 family redistribute in cells that have been exposed to stress. As such, the hsp70 Ssa4p of the budding yeast S. cerevisiae accumulates in nuclei when cells are treated with ethanol, whereas classical nuclear import is inhibited under these conditions. The N-terminal domain of Ssa4p, which is lacking a classical NLS, mediates nuclear accumulation upon ethanol exposure. Concentration of the Ssa4p N-terminal segment in nuclei is reversible, as the protein relocates to the cytoplasm when cells recover. Mutant analysis demonstrates that the small GTPase Gsp1p and GTPase-modulating factors are required to accumulate Ssa4p in nuclei upon ethanol stress. Moreover, we have identified the importin-beta family member Nmd5p as the nuclear carrier for Ssa4p. Ethanol treatment significantly increases the formation of import complexes containing Nmd5p and the N-terminal Ssa4p domain. Likewise, docking of the carrier Nmd5p at the nuclear pore is enhanced by ethanol. Furthermore, we show that the stressed-induced nuclear accumulation of Ssa4p depends on signaling through protein kinase C and requires sensors of the cell integrity pathway.


Swim pacemakers in box jellyfish are modulated by the visual input.
A major part of the cubozoan central nervous system is situated in the eye-bearing rhopalia. One of the neuronal output channels from the rhopalia carries a swim pacemaker signal, which has a one-to-one relation with the swim contractions of the bell shaped body. Given the advanced visual system of box jellyfish and that the pacemaker signal originates in the vicinity of these eyes, it seems logical to assume that the pacemakers are modified by the visual input. Here, the firing frequency and distribution of inter-signal intervals (ISIs) of single pacemakers are examined in the Caribbean box jellyfish, Tripedalia cystophora. It is shown that the absolute ambient light intensity, if kept constant, has no influence on the signal, but if the intensity changes, it has a major impact on both frequency and ISIs. If the intensity suddenly drops there is an increase in firing frequency, and the ISIs become more homogeneously distributed. A rise in intensity, on the other hand, produces a steep decline in the frequency and makes the ISIs highly variable. These electrophysiological data are correlated with behavioral observations from the natural habitat of the medusae.