Total laparoscopic and thoracoscopic Ivor Lewis esophagectomy after neoadjuvant Chemoradiation with minimal overall and anastomotic complications.
The published rates of morbidity and mortality remain relatively high for patients who undergo laparoscopic and thoracoscopic Ivor Lewis esophagectomy. We report the postoperative and oncologic outcomes of a large cohort of patients with esophageal carcinoma who were uniformly treated with laparoscopic and thoracoscopic Ivor Lewis esophagectomy following neoadjuvant chemoradiation. This is a retrospective observational study of 112 patients diagnosed with esophageal carcinoma who underwent total laparoscopic and thoracoscopic Ivor Lewis esophagectomy from May 2014 to May 2018. All of the patients received neoadjuvant chemoradiation consisting of 45 to 50.4 Gray of radiation and 3-5 cycles of carboplatin and paclitaxel chemotherapy. Perioperative morbidity and 90-day mortality were recorded. The overall and disease-free survival rates were estimated by Kaplan Meier techniques. A total of 112 patients completed induction chemoradiation followed by a total laparoscopic and thoracoscopic Ivor Lewis esophagectomy. There were 87 (77.68%) males and 25 (22.32%) females with a mean age of 61.6 years ± 10.4. A total of 28 (25%) patients had one or more complications. A total of 4 patients (3.57%) had an anastomotic leak. The 90-day mortality rate was 0.89%. The 3-year overall survival rate was 64.7% and the 3-year disease-free survival rate was 70.2%. The current outcomes suggest that laparoscopic and thoracoscopic Ivor Lewis esophagectomy can be performed with minimal overall and anastomotic complications following neoadjuvant chemoradiation.


Bone density, turnover, and estimated strength in postmenopausal women treated with odanacatib: a randomized trial.
Odanacatib, a cathepsin K inhibitor, increases spine and hip areal bone mineral density (BMD) in postmenopausal women with low BMD and cortical thickness in ovariectomized monkeys. The objective of the study was to examine the impact of odanacatib on the trabecular and cortical bone compartments and estimated strength at the hip and spine. This was a randomized, double-blind, 2-year trial. The study was conducted at a private or institutional practice. PARTICIPANTS included 214 postmenopausal women with low areal BMD. The intervention included odanacatib 50 mg or placebo weekly. Changes in areal BMD by dual-energy x-ray absorptiometry (primary end point, 1 year areal BMD change at lumbar spine), bone turnover markers, volumetric BMD by quantitative computed tomography (QCT), and bone strength estimated by finite element analysis were measured. Year 1 lumbar spine areal BMD percent change from baseline was 3.5% greater with odanacatib than placebo (P < .001). Bone-resorption marker C-telopeptide of type 1 collagen was significantly lower with odanacatib vs placebo at 6 months and 2 years (P < .001). Bone-formation marker procollagen I N-terminal peptide initially decreased with odanacatib but by 2 years did not differ from placebo. After 6 months, odanacatib-treated women had greater increases in trabecular volumetric BMD and estimated compressive strength at the spine and integral and trabecular volumetric BMD and estimated strength at the hip (P < .001). At the cortical envelope of the femoral neck, bone mineral content, thickness, volume, and cross-sectional area also increased from baseline with odanacatib vs placebo (P < .001 at 24 months). Adverse experiences were similar between groups. Over 2 years, odanacatib decreased bone resorption, maintained bone formation, increased areal and volumetric BMD, and increased estimated bone strength at both the hip and spine.


Effects of elicitors on tropane alkaloids and gene expression in Atropa baetica transgenic hairy roots.
Elicitation of transgenic Atropa baetica overexpressing the h6h gene with salicylic acid (SA), acetylsalicylic acid (ASA),or methyl jasmonate (MeJ) was conducted to boost tropane alkaloid yields. Scopolamine (1) amounts increased after treatment with ASA and MeJ, but not with SA. The highest enhancement of 1 was achieved with MeJ followed by ASA dissolved in EtOH. Transcriptomic analyses showed a direct relationship between content of 1 and gene expressions;the engineered h6h gene and other biosynthetic genes were stimulated. ASA dissolved in EtOH showed a high h6h gene expression, increasing 25-fold and 5-fold compared to controls; tr-I also displayed a 5-fold increase. The controls to which EtOH was added showed a 5-fold increase in h6h gene expression and 125-fold for pmt, demonstrating that EtOH also functioned as an enhancer of 1. MeJ was the best elicitor, displaying a 25-fold increase in h6h expression level, not affecting the expression of the other three genes analyzed, and it appears to possibly stimulate the phenylpropanoids branch of the tropane alkaloid pathway.


Public Knowledge of Osteoarthritis in Al-Qunfudah Governorate, Saudi Arabia.
Joint pain is one of the most frequent complaints among adults and older people in primary healthcare settings worldwide. There are many causes for joint pain, osteoarthritis (OA) is so far the most prevalent form of arthritis that causes joint pain. It can attack almost any joint, but the most frequently affected joints are the hands, knees, hips, and spine. This study aimed to identify public knowledge of OA and its associative variables in Al-Qunfudah governorate, Saudi Arabia. A cross-sectional descriptive community-based study was carried out among the general population in the Al-Qunfudah governorate. The research data were collected over two months, from November to December 2022, via an Arabic version of a self-administrated online survey of 29 items. A total of 746 respondents were included in this study. The majority of them were females (78%). The age group 18-29 was predominant. In terms of education, 69.9% were holding university degrees. The overall participants' knowledge of OA was poor at 36.1%, fair at 36.8%, and good at 26.9%. The associative variables with better participants' knowledge were; holding university degrees (P=0.021), being a student (P<0.001) and living in urban areas (P=0.020), having normal BMI (P=0.018), and depending on the school topics as a source of information (P<0.001). Good knowledge was significantly higher among healthy individuals and non-smokers (P<0.001) for each variable. This study reveals the lack of knowledge of osteoarthritis among the general population in Al-Qunfudah governorate, Saudi Arabia. Being a student, university educated, from urban areas, and having a normal BMI, all were associative factors with good knowledge. Therefore, this study highlights the necessity for providing awareness and educational campaigns for the public, focusing on the rural population.


Roles and mechanism of miR-199a and miR-125b in tumor angiogenesis.
MicroRNAs (miRNAs) have been shown to be involved in different aspects of cancer biology including tumor angiogenesis. In this study, we identified that two miRNAs, miR-199a and miR-125b were downregulated in ovarian cancer tissues and cell lines. Overexpression of miR-199a and miR-125b inhibited tumor-induced angiogenesis associated with the decrease of HIF-1α and VEGF expression in ovarian cancer cells. Moreover, the levels of miR-199a and miR-125b were negatively correlated with VEGF mRNA levels in ovarian tissues. We further showed that direct targets of miR-199a and miR-125b HER2 and HER3 were functionally relevant. Forced expression of HER2 and HER3 rescued miR-199a- and miR-125b-inhibiting angiogenesis responses and Akt/p70S6K1/HIF-1α pathway. This study provides a rationale for new therapeutic approach to suppress tumor angiogenesis using miR-199a, miR-125b, or their mimics for ovarian cancer treatment in the future.


Influence of the linkage type between the polymer backbone and side groups on the surface segregation of methyl groups during film formation.
Although poly(vinyl acetate) (PVAc) differs from poly(methyl acrylate) (PMA) only in the reversed position of the ester group, a large difference in the concentration dependence of the casting solution on the corresponding surface structure of the cast films of PVAc, PMA and poly(methyl methacrylate) (PMMA) was observed. The hydrophobicity of both PMA and PMMA films increased with increasing concentration of the corresponding polymer solution, whereas cast PVAc films showed the reverse trend. The surface structure of the cast films prepared with different concentrations of the casting solution, characterized by sum frequency generation (SFG) vibrational spectra, showed that the order of the methylene groups increased while that of the acetyl methyl group decreased on the surface of cast PVAc film with increasing concentration of casting solution. However, the order of the ester methyl group increased and that of methylene groups did not change for cast PMA films with increasing concentration of casting solution. The cast PMMA film showed a reverse trend compared with the corresponding PMA film. It is apparent that well-ordered ester or acetyl methyl groups on the surface, which are oriented away from the polymer film, rather than methylene groups, play an important role in determining surface hydrophobicity, as the latter shield the OC[double bond, length as m-dash]O groups of PVAc, PMA and PMMA film surfaces from being exposed, resulting in low surface free energy. The reason for this difference is attributed to the relatively low energy for ester methyl group reorientation, an ester group structure nearer to the trans state and more regular local configuration of segments in concentrated solutions of PMA and PMMA compared to that of PVAc.


New Polyketides With Anti-Inflammatory Activity From the Fungus Aspergillus rugulosa.
Two new polyketide compounds, asperulosins A and B (1-2), and one new prenylated small molecule, asperulosin C (3), along with nine known compounds (4-12), were isolated and identified from a fungus Aspergillus rugulosa. Their structures were extensively elucidated via HRESIMS, 1D, and 2D NMR analysis. The absolute configurations of the new compounds were determined by the comparison of their electronic circular dichroism (ECD), calculated ECD spectra, and the detailed discussion with those in previous reports. Structurally, compounds 1 and 2 belonged to the polyketide family and were from different origins. Compound 2 was constructed by five continuous quaternary carbon atoms, which occur rarely in natural products. All of the isolates were evaluated for anti-inflammatory activity against the production of nitric oxide (NO) in lipopolysaccharide (LPS)-induced RAW264.7 cells. Among those, compounds 1 and 5 showed a significant inhibitory effect on NO production with IC50 values of 1.49 ± 0.31 and 3.41 ± 0.85 μM, respectively. Additionally, compounds 1 and 5 markedly increased the secretion of anti-inflammatory cytokine IL10 while suppressing the secretion of pro-inflammatory cytokines IL6, TNF-α, IFN-γ, MCP-1, and IL12. Besides, 1 and 5 inhibited the transcription level of pro-inflammatory macrophage markers IL6, IL1β, and TNF-α while remarkably elevating the anti-inflammatory factor IL10 and M2 macrophage markers ARG1 and CD206. Moreover, 1 and 5 restrained the expression and nuclear translocation of NF-κB, as well as its downstream signaling proteins COX-2 and iNOS. All these results suggest that 1 and 5 have potential as anti-inflammatory agents, with better or comparable activities than those of the positive control, dexamethasone.


Complete Genome Sequence of Sphingopyxis macrogoltabida Strain 203N (NBRC 111659), a Polyethylene Glycol Degrader.
We determined the complete genome sequence of Sphingopyxis macrogoltabida strain 203N, a polyethylene glycol degrader. Because the PacBio assembly (285× coverage) seemed to be full of nucleotide-level mismatches, the Newbler assembly of MiSeq mate-pair and paired-end data was used for finishing and the PacBio assembly was used as a reference. The PacBio assembly carried 414 nucleotide mismatches over 5,953,153 bases of the 203N genome.


Radioiodide uptake in brain, CSF, thyroid, and salivary glands of audiogenic seizure mice.
DBA/2J (DBA) mice are susceptible to audiogenic seizures (ASs) in an age-dependent manner. Anion transport as measured by radioiodide uptake was determined in thyroid gland, salivary gland, skeletal muscle, cerebral cortex, cerebellum, brainstem, and CSF from these mice at various ages. Anion transport was also determined in C57BL/6J(C57) mice, an AS-resistant strain. In thyroid, DBA mice had an enhanced ability to concentrate iodide at 21 days of age when they have maximal AS susceptibility, as compared with the same-aged C57 mice. This difference in thyroid function was less marked at 40 days of age, when DBA mice are less AS susceptible, and was absent at 110 days of age, when DBA mice are AS resistant. In brain, differences in iodide uptake were also noted between these two strains of mice at 21 days of age. DBA mice had an increased concentration of iodide in CSF, an indication that they have a defect in the transport of iodide out of the CSF across the choroid plexus. In addition, DBA mice had a lower ratio of cerebral cortex to CSF iodide, which suggests that DBA mice have a defect in the transport of this anion into cerebral cortical cells from brain interstitial fluid. These differences in iodide transport in brain decreased with age as the AS susceptibility of DBA mice decreased. These results suggest a relation between anion transport in thyroid gland, cerebral cortex, and choroid plexus and AS susceptibility in DBA mice at 21 days of age.


[The white coat in pediatrics: link between medical history and preference for informally dressed physicians].
To find out if children show a preference for a doctor in a white coat or an informally dressed doctor, and what factors influence this preference. Further to determine if children show a preference for a physician of their own or of the other sex. A series of 20 pairs of photographs showing varying combinations of a male and a female doctor in a white coat or in informal dress was presented to test subjects: 37 children without a medical history, 28 children with a minor and 16 with an extensive medical history, aged between 6 and 12 years. Out of each pair of photographs, the test subjects were invited to choose the doctor they would prefer. On average, the test subjects opted for a doctor in a white coat more often than for an informally dressed doctor (p < 0.05). There was a significant difference in dress preference between children with a less and with a more extensive medical history (p = 0.001): children with no medical history more often chose a doctor in a white coat. Children with an extensive medical history more frequently showed preference for informal dress. The children opted more frequently for a doctor of their own sex (p < 0.001). The preference of children for a doctor in a white coat or for an informally dressed doctor depended on their medical history. The more extensive the medical history, the more the preference shifted to an informally dressed doctor. Also, the children preferred a doctor of their own sex.


Canadian Treatment Guidelines for Individuals at Clinical High Risk of Psychosis.
Young people who are at clinical high risk (CHR) of developing psychosis are often help seeking and have significant distress and dysfunction. There are limited guidelines for the assessment and treatment for this population. The aim of this guideline was to develop treatment recommendations for this at-risk group. A systematic search was conducted for published guidelines for CHR. All current guidelines for schizophrenia were reviewed for treatment guidelines on individuals at CHR. The recommendations adopted were primarily drawn from the European Psychiatric Association (EPA) guidance on the early intervention in clinical high-risk states of psychoses and the 2014 National Institute for Health and Care Excellence (NICE) guidelines on the treatment and management of those at CHR for psychosis. After the guideline development process described, 9 recommendations were developed based on the quality of evidence, appropriateness for the Canadian health care system, and clinical expert consensus. Assessment by an expert in the field was the first recommendation. It was recommended that treatment follow a staged approach with psychological treatments being the first-line treatment and pharmacotherapy reserved for adults, those who did not respond to psychological interventions, and those who had more severe symptoms.


Activation of the ribosomal DNA promoter in cells exposed to insulinlike growth factor I.
We constructed a stable cell line, 3T3A5, which carried a chimeric gene in which the simian virus 40 T-antigen-coding gene was under the control of the mouse ribosomal DNA promoter. These cells expressed T antigen when they were growing exponentially in 10% fetal calf serum, but they all became T negative when incubated for 5 days in low-concentration serum. The readdition of serum or platelet-poor plasma again induced the expression of T antigen, which was accompanied by an increase in steady-state levels of the corresponding RNA. Among the various growth factors tested for their ability to induce T-antigen expression in 3T3A5 cells, only insulinlike growth factor I (IGF-I) could induce T antigen at physiological concentrations. The effect of IGF-I or platelet-poor plasma was abolished by an antibody to IGF-I. Other growth factors, like insulin and epidermal growth factor, could induce the expression of T antigen in 3T3A5 cells, but only at concentrations far above the physiological range. Other growth factors were totally ineffective. These results indicate that exposure of cells to IGF-I can activate transcription from the ribosomal DNA promoter.


Epidural spinal infection in intravenous drug abusers.
The incidence of spinal epidural abscess and disk space infection appears to be rising in intravenous drug-using patients. We report 18 cases seen over three years in two municipal hospitals. Staphylococcus aureus was the most common infective agent, but two patients had Mycobacterium tuberculosis infection. Computed tomography of the spine facilitates diagnosis. Early treatment improves outcome but host factors play a role in recovery.


Co-catalyzed highly selective C(sp3)-H nitration.
A Co-catalyzed highly chemo- and regio-selective nitration of C(sp3)-H was developed. Diverse aliphatic nitro compounds were obtained in good yields, using t-BuONO as a nitrating reagent. Specific nitration of C(sp3)-H instead of C(sp2)-H was achieved via a radical process rather than concerted metalation-deprotonation.


Taxol resistance and its reversal by synthetic isoprenoids in human bladder cancer cell line.
We investigated the mechanism of action of reversal agents for taxol-resistance in bladder cancers. We isolated a taxol-resistant cell line (KK47/TX30) from a human KK47 bladder cancer cell line (KK47/WT). We characterized KK47/TX30 cells and screened reversal agents for taxol-resistance. KK47/TX30 cells exhibited approximately 700-fold resistance to taxol and cross-resistance to Vinca alkaloids and topoisomerase II inhibitors. Western blot analysis demonstrated P-glycoprotein (P-gp) overexpression in taxol-resistant cells. Drug accumulation and efflux studies showed that the decreased taxol accumulation in the resistant cell line was due to enhanced taxol efflux. We synthesized 31 isoprenoids based on the structure of N-solanesyl-N,N'-bis(3,4-dimethoxybenzyl)ethylenediamine (SDB), which could completely reverse multidrug resistance (MDR) as shown previously. Among those examined, trans-N,N'-bis(3,4-dimethoxybenzyl)-N-solanesyl-1,2-diaminocyclohexane (N-5228) could completely reverse taxol-resistance in KK47/TX30 cells. Results of a structure-activity relationship study of isoprenoids suggested that the following structural features were important for overcoming taxol-resistance: (1) a basic structure of 8 to 10 isoprene units, (2) a cyclohexane ring or benzene ring within the framework, (3) two cationic sites in close proximity to each other, and (4) a benzyl group with 3,4-dimethoxy functionalities with moderate electron-donation. Taxol-resistance was primarily mediated by P-gp overexpression in KK47/TX30 cells. One of the synthetic isoprenoids, N-5228 could completely reverse taxol-resistance in KK47/TX30 cells.


Improved Capacitive Energy Storage Nanocomposites at High Temperature Utilizing Ultralow Loading of Bimetallic MOF.
It is urgent to develop high-temperature dielectrics with high energy density and high energy efficiency for next-generation capacitor demands. Metal-organic frameworks (MOFs) have been widely used due to their structural diversity and functionally adaptable properties. Doping of metal nodes in MOFs is an effective strategy to change the band gap and band edge positions of the original MOFs, which helps to improve their ability to bind charges as traps. In this work, the incorporation of ultralow loading (<1.5 wt%) of novel bimetallic MOFs (ZIF 8-67) into the polyetherimide (PEI) polymer matrix is exhibited. With the addition of ZIF 8-67, the breakdown strength and energy storage capacity of ZIF 8-67/PEI nanocomposites are significantly improved, especially at high temperatures (200 °C). For example, the energy densitiy of the 0.5 wt% ZIF 8-67/PEI nanocomposite is up to 2.96 J cm-3 , with an efficiency (η) > 90% at 150 °C. At 200 °C, the discharge energy density of 0.25 wt% ZIF 8-67/PEI nanocomposites can still reach 1.84 J cm-3 with a η > 90%, which is nine times higher than that of pure PEI (0.21 J cm-3 ) under the same conditions, and it is the largest improvement compared with the previous reports.


Effects of carbon dioxide on Penicillium chrysogenum: An autoradiographic study.
Previous research has shown that dissolved carbon dioxide causes significant changes in submerged penicillin fermentations, such as stunted, swollen hyphae, increased branching, lower growth rates, and lower penicillin productivity. Influent carbon dioxide levels of 5 and 10% were shown through the use of autoradiography to cause an increase in chitin synthesis in submerged cultures of Penicillium chrysogenum. At an influent 5% carbon dioxide level, chitin synthesis is ca. 100% greater in the subapical region of P. chrysogenum hyphae than that of the control, in which there was no influent carbon dioxide. Influent carbon dioxide of 10% caused an increase of 200% in chitin synthesis, it is believed that the cell wall must be plasticized before branching can occur and that high amounts of dissolved carbon dioxide cause the cell to lose control of the plasticizing effect, thus the severe morphological changes occur.


The kinase domain of Jak2 mediates induction of bcl-2 and delays cell death in hematopoietic cells.
Granulocyte-macrophage colony-stimulating factor (GM-CSF), interleukin (IL)-3, and IL-5 stimulate DNA synthesis and proliferation and inhibit apoptosis in hematopoietic cells. Multiple signal pathways are activated by binding of these ligands to their receptors, which share a common beta subunit. Janus protein kinase 2 (Jak2) binds to the membrane proximal domain of the beta chain and is phosphorylated on receptor ligation. To explore the role of Jak2 in the regulation of specific signal transduction pathways, we constructed fusion proteins with a CD16 external domain, a CD7 transmembrane region, and a Jak2 cytoplasmic domain. This cytoplasmic domain consisted either of wild type Jak2 (CD16/Jak2-W) or Jak2 mutations with deletions of (a) the amino terminus (CD16/Jak2-N), (b) kinase-like domain (CD16/Jak2-B), (c) kinase domain (CD16/Jak2-C), or (d) amino-terminal and kinase-like domains, leaving the kinase domain (CD16/Jak-K) intact. In contrast to the CD16/Jak2-W fusion protein, which requires cross-linking for activation, CD16/Jak2-N, CD16/Jak2-B, and CD16/Jak2-K were constitutively phosphorylated, and they stimulated Shc phosphorylation and increased binding of STAT to DNA in Ba/F3 cells. Cell lines derived from IL-3-dependent Ba/F3 cells stably transfected with CD16/Jak2-W, CD16/Jak2-N, or CD16/Jak2-B mammalian expression vectors died at a rate similar to that of the parental cells on IL-3 deprivation. In contrast, CD16/Jak2-K cell lines exhibited increased expression of bcl-2 and pim-1 mRNA and maintained their viability when compared with control cell lines. Thus, activation of tyrosine phosphorylation by creating a CD16/Jak2-K fusion is sufficient to activate pathways that prevent cell death.


Policies to Reduce Lead Exposure: Lessons from Buffalo and Rochester.
Lead exposure remains a major issue in cities, such as Buffalo and Rochester, with concentrated, segregated poverty and old, deteriorated housing stock. Exploring and comparing local policies and programs in these two cities, the author suggests that increasing the number of proactive housing inspections in high-risk areas and forming a single-purpose non-profit group dedicated to lead education and advocacy are two valuable interventions. He recommends additional policy steps, such as more stringent inspection standards; state adoption of the Environmental Protection Agency's Renovation and Repair Program; the lowering of state elevated blood level thresholds; a focus on in-person, interactive education by community health workers; and more vigorous enforcement of testing requirements among physicians.


Effect of Short Glass Fiber Addition on Flexural and Impact Behavior of 3D Printed Polymer Composites.
Fused deposition modeling (FDM), one of the most widely used additive manufacturing (AM) processes, is used for fabrication of 3D models from computer-aided design data using various materials for a wide scope of applications. The principle of FDM or, in general, AM plays an important role in minimizing the ill effects of manufacturing on the environment. Among the various available reinforcements, short glass fiber (SGF), one of the strong reinforcement materials available, is used as a reinforcement in the acrylonitrile butadiene styrene (ABS) matrix. At the outset, very limited research has been carried out till date in the analysis of the impact and flexural strength of the SGF-reinforced ABS polymer composite developed by the FDM process. In this regard, the present research investigates the impact and flexural strength of SGF-ABS polymer composites by the addition of 15 and 30 wt % SGF to ABS. The tests were conducted as per ASTM standards. Increments in flexural and impact properties were observed with the addition of SGF to ABS. The increment of 42% in impact strength was noted for the addition of 15 wt % SGF and 54% increase with the addition of 30 wt % SGF. On similar lines, flexural properties also showed improved values of 44 and 59% for the addition of 15 and 30 wt % SGF to ABS. SGF addition greatly enhanced the properties of flexural and impact strength and has paved the path for the exploration of varied values of reinforcement into the matrix.


Retention of Microbiota Diversity by Lactose-Free Milk in a Mouse Model of Elderly Gut Microbiota.
Prebiotics may improve aging-related dysbiosis. Milk is a source of nutrients including oligosaccharides whose prebiotic potential remains largely unexplored. We used a murine model to explore the effect of milk products on high diversity and lower diversity faecal microbiota from healthy and frail elderly subjects, respectively. Mice were treated with antibiotics and subsequently "humanized" with human faecal microbiota. The mice received lactose-free or whole milk, glycomacropeptide, or soy protein (control) supplemented diets for one month. The faecal microbiota was analyzed by 16S rRNA gene amplicon sequencing. Lactose-free milk diet was as efficient as the control diet in retaining faecal microbiota diversity in mice. Both milk diets had a significant effect on the relative abundance of health-relevant taxa (e.g., Ruminococcaceae, Lachnospiraceae). The glycomacropeptide prebiotic activity previously observed in vitro was not replicated in vivo. However, these data indicate the novel prebiotic potential of bovine milk for human nutrition.


Photochemically induced cataracts in rat lenses can be prevented by AL-3823A, a glutathione peroxidase mimic.
Oxidative stress is known to cause cataracts in lens culture systems and is believed to be an important factor contributing to human cataracts. In this communication, it is demonstrated that cataract development of cultured rat lenses produced as a result of photochemically induced oxidation in a 4% oxygen atmosphere similar to the native environment of the lens can be blocked by the transition metal complex AL-3823A. In this system, riboflavin is added to the medium as a photosensitizer. AL-3823A acts primarily as a glutathione peroxidase mimic, which catalytically metabolizes H2O2 and also has low superoxide dismutase-like activity. Measurements of H2O2, O2.-, and OH. indicate that appreciable levels of the first two of these oxidants and low levels of OH. are produced by this photochemical stressing system. The H2O2 concentrations are similar to those found in some patients with cataracts. The development of cataracts was followed over a 96-hr period. Transparency, hydration, glyceraldehyde-3-phosphate dehydrogenase activity, and protein and nonprotein thiol were monitored. All parameters show marked changes during the 96-hr period. However, in the presence of 200 microM AL-3823A, no difference between control and light-exposed lenses was observed with respect to these parameters. The results suggest that in vivo human cataract development caused by oxidative stress may be prevented by compounds of this type.


Black Phosphorus Nanosheets Immobilizing Ce6 for Imaging-Guided Photothermal/Photodynamic Cancer Therapy.
In preclinical and clinical research, to destroy cancers, particularly those located in deep tissues, is still a great challenge. Photodynamic therapy and photothermal therapy are promising alternative approaches for tissue cancer curing. Black phosphorus (BP)-based nanomaterials, with broad UV-vis near-infrared absorbance and excellent photothermal effect, have shown great potential in biomedical applications. Herein, a biocompatible therapeutic platform, chlorin e6 (Ce6)-decorated BP nanosheets (NSs), has been developed for fluorescence and thermal imaging-guided photothermal and photodynamic synergistic cancer treatment. Taking advantage of the relatively high surface area of exfoliated BP NSs, the PEG-NH2-modified BP NSs (BP@PEG) are loaded with a Ce6 photosensitizer. The resulted BP@PEG/Ce6 NSs not only have good biocompatibility, physiological stability, and tumor-targeting property but also exhibit enhanced photothermal conversion efficiency (43.6%) compared with BP@PEG NSs (28.7%). In addition, BP@PEG/Ce6 NSs could efficiently generate reactive oxygen species because of the release of the Ce6 photosensitizer, which is also verified by in vitro studies. In vivo fluorescence imaging suggests that BP@PEG/Ce6 NSs can accumulate in the tumor targetedly through the enhanced permeability and retention effect. Both in vitro and in vivo studies suggest that BP@PEG/Ce6 can be a promising nanotheranostic agent for synergetic photothermal/photodynamic cancer therapy.


Evidence of aerosols as a media for rapid daytime HONO production over China.
Current knowledge of daytime HONO sources remains incomplete. A large missing daytime HONO source has been found in many places around the world, including polluted regions in China. Conventional understanding and recent studies attributed this missing source mainly to ground surface processes or gas-phase chemistry, while assuming aerosols to be an insignificant media for HONO production. We analyze in situ observations of HONO and its precursors at an urban site in Beijing, China, and report an apparent dependence of the missing HONO source strength on aerosol surface area and solar ultraviolet radiation. Based on extensive correlation analysis and process-modeling, we propose that the rapid daytime HONO production in Beijing can be explained by enhanced hydrolytic disproportionation of NO2 on aqueous aerosol surfaces due to catalysis by dicarboxylic acid anions. The combination of high abundance of NO2, aromatic hydrocarbons, and aerosols over broad regions in China likely leads to elevated HONO levels, rapid OH production, and enhanced oxidizing capacity on a regional basis. Our findings call for attention to aerosols as a media for daytime heterogeneous HONO production in polluted regions like Beijing. This study also highlights the complex and uncertain heterogeneous chemistry in China, which merits future efforts of reconciling regional modeling and laboratory experiments, in order to understand and mitigate the regional particulate and O3 pollutions over China.


Blood-brain barrier disruption results in delayed functional and structural alterations in the rat neocortex.
Disruption of the blood-brain barrier (BBB) is a characteristic finding in common neurological disorders. Human data suggest BBB disruption may underlie cerebral dysfunction. Animal experiments show the development of epileptiform activity following BBB breakdown. In the present study we investigated the neurophysiological, structural and functional consequences of BBB disruption. Adult rats underwent focal BBB disruption in the rat sensory-motor cortex using the bile salt sodium deoxycholate (DOC). Magnetic resonance imaging in-vivo showed an early BBB disruption with delayed reduction in cortical volume. This was associated with a reduced number of neurons and an increased number of astrocytes. In-vitro experiments showed that the threshold for spreading depression and the propagation velocity of the evoked epileptic potentials were increased 1 month after treatment. Furthermore, animals' motor functions deteriorated during the first few weeks following BBB disruption. Treatment with serum albumin resulted in a similar cell loss confirming that the effect of DOC was due to opening of the BBB. Our findings suggest that delayed neurodegeneration and functional impairment occur following the development of the epileptic focus in the BBB-permeable cerebral cortex.


A combination of oral sildenafil and beraprost ameliorates pulmonary hypertension in rats.
Sildenafil, an oral phosphodiesterase type-5 inhibitor, has vasodilatory effects through a cyclic guanosine 3', 5'-monophosphate-dependent mechanism, whereas beraprost, an oral prostacyclin analog, induces vasorelaxation through a cAMP-dependent mechanism. We investigated whether the combination of oral sildenafil and beraprost is superior to each drug alone in the treatment of pulmonary hypertension. Rats were randomized to receive repeated administration of saline, sildenafil, beraprost, or both of these drugs twice a day for 3 weeks. Three weeks after monocrotaline (MCT) injection, there was significant development of pulmonary hypertension. The increases in right ventricular systolic pressure and ratio of right ventricular weight to body weight were significantly attenuated in the Sildenafil and Beraprost groups. Combination therapy with sildenafil and beraprost had additive effects on increases in plasma cAMP and cyclic guanosine 3', 5'-monophosphate levels, resulting in further improvement in pulmonary hemodynamics compared with treatment with each drug alone. Unlike MCT rats given saline, sildenafil, or beraprost alone, all rats treated with both drugs remained alive during 6-week follow-up. These results suggest that combination therapy with oral sildenafil and beraprost attenuates the development of MCT-induced pulmonary hypertension compared with treatment with each drug alone.


ATF3 Positively Regulates Antibacterial Immunity by Modulating Macrophage Killing and Migration Functions.
The clinical severity of Staphylococcus aureus (S. aureus) respiratory infection correlates with antibacterial gene signature. S. aureus infection induces the expression of an antibacterial gene, as well as a central stress response gene, thus activating transcription factor 3 (ATF3). ATF3-deficient mice have attenuated protection against lethal S. aureus pneumonia and have a higher bacterial load. We tested the hypothesis that ATF3-related protection is based on the increased function of macrophages. Primary marrow-derived macrophages (BMDM) were used in vitro to determine the mechanism through which ATF3 alters the bacterial-killing ability. The expression of ATF3 correlated with the expression of antibacterial genes. Mechanistic studies showed that ATF3 upregulated antibacterial genes, while ATF3-deficient cells and lung tissues had a reduced level of antibacterial genes, which was accompanied by changes in the antibacterial process. We identified multiple ATF3 regulatory elements in the antibacterial gene promoters by chromatin immunoprecipitation analysis. In addition, Wild type (WT) mice had higher F4/80 macrophage migration in the lungs compared to ATF3-null mice, which may correlate with actin filament severing through ATF3-targeted actin-modifying protein gelsolin (GSN) for the macrophage cellular motility. Furthermore, ATF3 positively regulated inflammatory cytokines IL-6 and IL-12p40 might be able to contribute to the infection resolution. These data demonstrate a mechanism utilized by S. aureus to induce ATF3 to regulate antibacterial genes for antimicrobial processes within the cell, and to specifically regulate the actin cytoskeleton of F4/80 macrophages for their migration.


Case of labial sclerosing polycystic adenoma with ductal carcinoma in situ (DCIS).
Minor salivary gland tumours are enigmatic in their behaviour and presentations. Histopathological diagnosis of these tumours encompasses a large spectrum. Conventionally, small nodular tumours of the upper lip or the labial mucosa are generally adenomas, frequently pleomorphic adenomas. Here, we describe a case of a solitary nodular tumour, occurring in the upper labial mucosa, diagnosed as sclerosing polycystic adenoma (SPA) with intraductal epithelial proliferation of high grade. This is a rare lesion, which has entered into the category of salivary gland tumours recently in the 2017 WHO categorisation. We report a case of paucicystic SPA with intraductal epithelial proliferations in the labial minor salivary gland of a 56-year-old woman, which might be the first report of a case occurring in the upper labial mucosa.


A Rare Case Report and Literature Review of Anabolic-Androgenic Steroids (AAS)-Induced Acute Myocardial Infarction.
Anabolic-androgenic steroids (AAS) abuse is common in competitive athletes in order to enhance athletic performances. However, AAS abuse is often associated with deleterious side effects including but not limited to cardiovascular diseases, depression, hormonal abnormalities, and cancer. We present a case of a 31-year-old male with a history of Crohn's disease on infliximab and chronic AAS use who had persistent retrosternal chest pain found to have an acute myocardial infarction (MI) without obvious cardiovascular risk factors.


[Surgical treatment of metastatic spinal malignancies].
The goal of surgical treatment of metastatic tumors of the spine is maintaining the relief of pain and/or neurologic symptoms for as long as possible after surgery. For the quality of life, thirty-five patients were surgically treated. By preoperative analyses, tumor invasion was divided into 3 types as follows; Type 1: involvement of vertebral body, Type 2: involvement of posterior column, Type 3: involvement of anterior and posterior column. Of 35 patients, 31 were in Type 1 or 3, who underwent anterior and/or posterior direct decompression by removal of the tumor, followed by vertebral reconstruction. Posterior surgery was performed on the remaining 4 patients in Type 2, resulting in complete decompression. A combined anterior or posterior instrumentation with anterior reconstruction for the involved vertebral body in Type 1 and 3 provided rigid spinal stability immediately after surgery. Neurologic improvement was evaluated by Frankel's classification. As a result, 4 patients who are alive have been ambulatory for an average duration of 26.3 months. Thirty-one, who survived for an average duration of 10 months, had no neurologic deterioration in an average duration of 8.4 months. Removal of the tumor and reconstructive surgery combined with rigid spinal instrumentation may provide satisfactory results.


State Medicaid and private payer reimbursement for telemedicine: An overview.
Individual states in the USA were given the option of paying for telemedicine services with Medicaid (i.e. federal health-care funds administered by the state) in 1998, when the Health Care Financing Administration (HCFA) published final rules for Medicare payment for teleconsultations in health professional shortage areas (HPSAs). It was left to telemedicine practitioners in each state to negotiate the scope of the services covered with the state Medicaid office. Three reports of data gathered by 2002-03 surveys on state reimbursement policies have been reviewed, with additional information from a brief informal 2005 survey conducted by the author. In the seven years since 1998, 34 states have added coverage of telemedicine services to their Medicaid programmes, although there are wide variations in service coverage, payment policies, and geographical and other restrictions. There is less published information on private payer reimbursement. One survey performed by AMD Telemedicine (AMD) and the American Telemedicine Association (ATA) showed that over half of the 72 telemedicine programmes in 25 states delivering billable services were being reimbursed by private payers. In 1999, 43% of responding telemedicine networks saw reimbursement as a barrier to long-term sustainability, while in 2004 only 22% did so. It appears that some progress has been made in Medicaid and private payer reimbursement for telemedicine.


Metabolic screen and intervene: improving mental health inpatient metabolic monitoring.
To assess rates of metabolic monitoring in patients prescribed antipsychotic medications in the psychiatric inpatient setting and the impact education can have regarding monitoring compliance. Two identical audits were undertaken at a NSW mental health inpatient service before and after a campaign designed to educate mental health workers about the importance of metabolic monitoring. Results from both audits were compared for statistically significant improvements in monitoring rates. Rates of monitoring plasma lipids increased from 21.7% to 78.8% (p < 0.01) and rates for plasma glucose increased from 20.8% to 73.7% (p < 0.01). There were no statistically significant changes in rates of monitoring body mass index (83.0% and 77.1%, respectively), waist circumference (36.8% and 43.2%, respectively) and blood pressure (99.1% and 100%, respectively). This study has shown that rates of metabolic monitoring in the inpatient setting can be improved with a relatively low-cost education intervention. While absolute rates remain low, outcomes suggest that it may be worthwhile trialling further modes of education and repeating this education in cycles.


Otitis media in the first year of life in two Eskimo communities.
Severe otitis media of early onset is particularly prevalent among Eskimo infants. An investigation of health records of all infants born in 1984 and served by two health facilities was undertaken to assess the relative significance of three particular variables associated with this problem: 1) sex versus age, 2) sex versus severity, and 3) sex versus other health conditions. Ninety-five percent of the children in one locale and 74.4% in the other had at least one episode during the first year of life. While none of the comparisons were statistically significant, infants at the northernmost community were generally more severely affected; females in each community were found to have 1) fewer and less severe episodes of disease, 2) the first episode at a later age, 3) fewer episodes during the year, and 4) fewer episodes of bilateral disease.


Serotonin: beyond the brain.
The Serotonin Club has played a crucial role in disseminating information on the roles played by this monoamine neurotransmitter, as well as in the process of pharmacological classification of serotonin membrane receptors. The Club's future mission should be to emphasize that serotonin (5-hydroxytyptamine) contributes to physiology and pathology beyond its function in the central nervous system.


Fat-soluble vitamin deficiency and exocrine pancreatic insufficiency among adults with chronic pancreatitis: Is routine monitoring necessary for all patients?
Chronic pancreatitis (CP) is often associated with exocrine pancreatic insufficiency (EPI), which may increase risk for fat-soluble vitamin depletion. Although vitamin D deficiency is widespread among the general population, vitamins A, E, and K deficiencies may more uniquely present in patients with CP. Yet, it is unclear whether fat-soluble vitamin status should be routinely monitored in all patients with CP or limited to those with EPI. The purpose of this review is to describe the laboratory status of vitamins A, E, and K in adult patients with CP and their association with exocrine pancreatic function. Five primary, observational studies met the inclusion criteria for qualitative synthesis. Biochemical deficiencies in fat-soluble vitamins were observed across trials but results varied with respect to whether EPI increased risk. Challenges related to the diagnosis and treatment of EPI along with potential confounders may contribute to the heterogeneity among study results. Although more studies are needed to determine the influence of pancreatic enzyme replacement therapy on fat-soluble vitamin status as well as effective vitamin repletion strategies, clinicians should consider periodically screening for deficiencies in all patients with CP regardless of EPI to avoid associated health effects of vitamin depletion.


Neoadjuvant and adjuvant chemotherapy for muscle-invasive bladder cancer: The likelihood of initiation and completion.
Chemotherapy was shown to improve survival in patients undergoing radical cystectomy (RC) for muscle-invasive bladder cancer (MIBC). The initiation and completion rates for perioperative chemotherapy are variable. Our aim is to compare the likelihood of initiating and completing neoadjuvant (NAC) and adjuvant chemotherapy (AC) in patients who underwent of RC for MIBC. We performed a retrospective analysis of patients who underwent RC between 1992 and 2011. NAC was advised for patients with clinical stage ≥T2, hydronephrosis, extensive lymphovascular invasion (LVI), or prostatic stromal invasion. Patients with ≥pT3 or lymph node metastases were considered for AC. A total of 363 patients were considered for perioperative chemotherapy. Among the 141 patients who were offered NAC, 125 (88.6%) initiated NAC. A total of 222 were considered for AC, and 151 (68.0%) initiated AC (P < 0.001). In the NAC group, 118 (83.5%) completed planned number of cycles of chemotherapy and 7 (5.6%) did not complete the planned chemotherapy. In the AC group, 79 (35.5%) completed at least four cycles and 72 (47.3%) could not complete the planned cycles (P < 0.001). Patients with MIBC are more likely to initiate and complete NAC than AC.


Infant attachment security and early childhood behavioral inhibition interact to predict adolescent social anxiety symptoms.
Insecure attachment and behavioral inhibition (BI) increase risk for internalizing problems, but few longitudinal studies have examined their interaction in predicting adolescent anxiety. This study included 165 adolescents (ages 14-17 years) selected based on their reactivity to novelty at 4 months. Infant attachment was assessed with the Strange Situation. Multimethod BI assessments were conducted across childhood. Adolescents and their parents independently reported on anxiety. The interaction of attachment and BI significantly predicted adolescent anxiety symptoms, such that BI and anxiety were only associated among adolescents with histories of insecure attachment. Exploratory analyses revealed that this effect was driven by insecure-resistant attachment and that the association between BI and social anxiety was significant only for insecure males. Clinical implications are discussed.


Convergence of isoenzyme expression in human tumor xenografts.
We have grown 7 bona fide non-HeLa contaminated cell lines in the cheek pouch of immunosuppressed adult hamsters. Six of the cell lines studied produced or continued to express the chorionic form of alkaline phosphatase under these conditions. The Regan isoenzyme produced in culture by the J82, T24, HCT-8, and COLO 16 cell lines disappeared in vivo with a concomitant appearance of the chorionic isoenzyme. The other cell lines, however, maintained the expression of the Regan isoenzyme when grown as a xenograft: the C41 line produced the Regan isoenzyme in culture, and continued to do so in vivo, while the BeWo cells, which synthesized the chorionic enzyme in culture were induced by in vivo growth to produce both the Regan and chorionic isoenzymes. No expression of the oncoamniotic (FL) isoenzyme was observed in any of the lines studied. Our results indicate a convergence of human tumor xenografts with respect to the production of the chorionic form of alkaline phosphatase. Furthermore, the loss of production of the Regan isoenzyme was seen in most, but not all cell lines studied. Since the oncoamniotic (FL) isoenzyme was only observed by HeLa cells growing as a xenograft, the expression of this enzyme form may be a HeLa specific phenomenon. We must now look to the regulation of the chorionic isoenzyme as the most promising avenue of approach to understanding the biological significance of altered isoenzyme expression in human tumor xenografts.


Treating TB human neuroectodermal cell line with retinoic acid induces the appearance of neuron-like voltage-gated ionic currents.
TB is a cell line derived from the cerebrospinal fluid sample of a patient with primary leptomeningeal melanomatosis. Our previous immunological and ultrastructural analysis revealed that TB cells differentiate towards a neuronal phenotype when grown in vitro up to 7 days in presence of 10 µM all-trans retinoic acid (RA). Recently, we reported that TB cells are sensitive to the cytotoxic effects of β-amyloid peptides, activating the cytosolic phospholipase A2. To date, it is not known if RA, in addition to inducing morphological changes, also causes functional modification in TB cells, by regulating voltage-gated ionic currents. To this purpose, we performed electrophysiological characterization of undifferentiated (TB) and differentiated (RA-TB) cells by means of whole-cell patch clamp recordings. Upon depolarizing stimuli, both groups displayed voltage-gated K+ outward currents of similar amplitude. By contrast, the low amplitude voltage-gated Na+ currents recorded in undifferentiated TB cells were largely up-regulated by RA exposure. This current was strongly reduced by TTX and lidocaine and completely abolished by removal of extracellular sodium. Furthermore, treatment with RA caused the appearance of a late-onset inward current carried by Ca2+ ions in a subpopulation of TB cells. This current was not affected by removal of extracellular Na+ and was completely blocked by Cd2+, a broad-spectrum blocker of Ca2+ currents. Altogether, our results indicate that RA-differentiation of TB cells induces functional changes by augmenting the amplitude of voltage-gated sodium current and by inducing, in a subpopulation of treated cells, the appearance of a voltage-gated calcium current.


Decay of Moloney leukemia virus production after enucleation of chronically infected mouse cells.
The effect of enucleation of synthesis of Moloney leukemia virus was studied in chronically infected YACIR-A9 mouse hybrid cells. The synthesis of infectious virus decreased gradually until, 12 h after enucleation, virtually no infectious virus was produced. As determined by particle-bound p30 and reverse transcriptase, cytoplasts produced a low level of noninfectious viral particles. Thus, the assembly of virus particles does not appear to be wholly dependent on the presence of the cell nucleus. In contrast, production of infectious particles shows an absolute requirement for the cell nucleus.


Cell-Based Therapies with T Regulatory Cells.
CD4+CD25highFoxP3+ T regulatory cells (Tregs) are immunodominant suppressors in the immune system. Tregs use various mechanisms to control immune responses. Preclinical data from animal models have confirmed the huge therapeutic potential of Tregs in many immune-mediated diseases. Hence, these cells are now on the road to translation to cell therapy in the clinic as the first clinical trials are accomplished. To date, clinical research has involved mainly hematopoietic stem cell transplantations, solid organ transplantations, and autoimmunity. Despite difficulties with legislation and technical issues, treatment is constantly evolving and may soon represent a valid alternative for patients with diseases that are currently incurable. This review focuses on the basic and clinical experience with Tregs with adoptive transfer of these cells, primarily from clinical trials, as well as on perspectives on clinical use and technical problems with implementing the therapy.


Energy and greenhouse gas balances for a solid waste incineration plant: a case study.
Energy and greenhouse gas balances for a waste incineration plant (Reno-Nord I/S, Aalborg, Denmark) as a function of time over a 45-year period beginning 1960 are presented. The quantity of energy recovered from the waste increased over time due to increasing waste production, increasing lower heating value of the waste and implementation of improved energy recovery technology at the incineration plant. Greenhouse gas (GHG) balances indicated progressively increasing GHG savings during the time period investigated as a result of the increasing energy production. The GHG balances show that the Reno-Nord incineration plant has changed from a net annual GHG emission of 30 kg CO(2)-eq person(-1) year(-1) to a net annual GHG saving of 770 kg CO(2)-eq person(-1) year(-1) which is equivalent to approximately 8% of the annual emission of GHG from an average Danish person (including emissions from industry and transport). The CO(2) emissions associated with combustion of the fossil carbon contained in the waste accounted for about two-thirds of the GHG turnover when no energy recovery is applied but its contribution reduces to between 10 and 15% when energy recovery is implemented. The reason being that energy recovery is associated with a large CO(2) saving (negative emission).


4-{2-[2-(4-Chloro-benzyl-idene)hydrazinyl-idene]-3,6-dihydro-2H-1,3,4-thia-diazin-5-yl}-3-(4-meth-oxy-phen-yl)sydnone.
The title compound, C(19)H(15)ClN(6)O(3)S, exists in trans and cis configurations with respect to the acyclic C=N bonds. The 3,6-dihydro-2H-1,3,4-thia-diazine ring adopts a half-boat conformation. The sydnone ring is approximately planar [maximum deviation = 0.013 (1) Å] and forms dihedral angles of 34.76 (4) and 48.67 (4)° with the benzene rings. An intra-molecular C-H⋯O hydrogen bond stabilizes the mol-ecular structure and forms an S(6) ring motif. In the crystal packing, inter-molecular N-H⋯N hydrogen bonds link centrosymmetrically related mol-ecules into dimers, generating R(2) (2)(8) ring motifs. The dimers are then linked into a three-dimensional network by inter-molecular C-H⋯O and C-H⋯Cl hydrogen bonds, and by C-H⋯π inter-actions. Further stabilization is provided by π-π inter-actions involving the sydnone rings, with centroid-centroid separations of 3.4198 (5) Å.


Retinal tear: an unusual complication of ocular toxoplasmosis.
It is aimed to report on a 16-year-old patient with acquired ocular toxoplasmosis complicated by a retinal tear. Retrospective medical chart review. A 16-year-old Caucasian female presented with vision loss in her right eye. In addition to a white active lesion between the fovea and the optic nerve head, marked vitreous opacification was noted. She was diagnosed with ocular toxoplasmosis. The patient was treated with oral azithromycin, clindamycin, and trimethoprimsulfamethoxazole. One month later, retinochoroiditis resolved and vitreous cleared. Three months after onset, patient presented with floaters in the right eye and a retinal tear was located at the temporal region of the retina. Prophylactic argon laser treatment that encircled the retinal tear was performed. No other abnormalities were noted during 6 months of follow-up. Retinal tear associated with ocular toxoplasmosis is rare; however, a retinal tear can occur due to vitreoretinal traction following post-inflammatory structural alteration of the vitreous. Retinal tears may be seen during the healing phase, when the inflammation turns into tightening of vitreous substance. Careful retinal examination in cases of ocular toxoplasmosis is warranted, especially in patients with severe vitreous inflammation.


X-linked mental retardation with dystonic movements of the hands (PRTS): revisited.
A family with a syndrome of mental retardation, dystonic movements of the hands, and dysarthria (MIM no. 309510) was described and mapped to Xp22 by Partington et al. (Am J Med Genet 1988; 30:251-262). The original localization encompassed the distal half of the short arm of the X chromosome, with a peak lod score of 2.1 at the DXS41 locus. The gene localization for this disorder (PRTS) has now been refined using recently characterized dinucleotide repeat markers. The PRTS gene maps between DXS365 and DXS28, an interval estimated to be less than 15 cM. A peak lod score of 3.01 at a recombination fraction of zero was generated by 2-point linkage analysis with the marker DXS989. Dystonic movements may be progressive and could be overlooked in children. Clinical assessments of affected men who are mentally retarded should be critically evaluated for this manifestation, where they belong to families in which the gene localization overlaps with PRTS.


Emerging monoclonal antibodies against Clostridium difficile infection.
Clostridium difficile infections are characterized by a high recurrence rate despite antibiotic treatments and there is an urgent need to develop new treatments such as fecal transplantation and immonotherapy. Besides active immunotherapy with vaccines, passive immunotherapy has shown promise, especially with monoclonal antibodies. Areas covered: Herein, the authors review the different assays performed with monoclonal antibodies against C. difficile toxins and surface proteins to treat or prevent primary or recurrent episodes of C. difficile infection in animal models and in clinical trials as well. Notably, the authors lay emphasis on the phase III clinical trial (MODIFY II), which allowed bezlotoxumab to be approved by the Food and Drug Administration and the European Medicines Agency. They also review new strategies for producing single domain antibodies and nanobodies against C. difficile and new approaches to deliver them in the digestive tract. Expert opinion: Only two human Mabs against TcdA and TcdB have been tested alone or in combination in clinical trials. However, many animal model studies have provided rationale for the use of Mabs and nanobodies in C. difficile infection and pave the way for further clinical investigation.


Potassium oxonate induces acute hyperuricemia in the tree shrew (tupaia belangeri chinensis).
Potassium oxonate, a selectively competitive uricase inhibitor, produced hyperuricemia (HUA) in rodents in a previous study. In this study, we employed the tree shrew as an animal model to study potassium oxonate-induced HUA. The effect of allopurinol (ALLO), a uric acid reducer, was also examined in this model. Potassium oxonate at doses of 5, 20, 40, 60, 80, 100, and 1,000 mg/kg was given intraperitoneally to tree shrews. The results showed that potassium oxonate can effectively increase the levels of uric acid in tree shrews at doses ranging from 40 to 100 mg/kg. Semiquantitative RT-PCR showed that the xanthine dehydrogenase/oxidase (XDH/XO) mRNA expression level was significantly higher in the liver tissue of tree shrews with high levels of uric acid. There were no changes in serum urea nitrogen, or serum creatinine values. ALLO can significantly decrease serum uric acid levels (P<0.01) and raise XDH/XO mRNA expression levels in the liver tissue of tree shrews with HUA. XDH/XO mRNA expression levels did not change in untreated tree shrews. Studies on acute toxicity in the tree shrew did not show any significantly abnormal signs. There were no adverse effects at the macroscopic level up to doses ≤100 mg/kg. Potassium oxonate induced acute HUA in tree shrews at lower doses compared with other animal models. Potassium oxonate-treated tree shrews may be a potential animal model for studying pathogenic mechanism and evaluating a new therapeutic agent for treatment of HUA in humans.


[Folic acid intake in daily food rations of 20-25 years old women from Wroclaw and assessment of knowledge about its importance for health].
The mean folic acid content in the diets of 50 young women from Wroclaw was 282 mcg and complied in 70.5% of recommended intake in prophylactic of neural tube defects. Only 8% of women used peri-conceptional folate supplementation every day. In the examined group low knowledge about importance of folic acid in health was noticed. Educational interventions are required to inform women at childbearing age of the benefits resulting from supplementation of diet with folic acid.


30 YEARS OF THE MINERALOCORTICOID RECEPTOR: Mineralocorticoid receptor and NaCl transport mechanisms in the renal distal nephron.
A key role of aldosterone and mineralocorticoid receptor is to regulate fluid volume and K+ homeostasis in the body by acting on the renal distal nephron. Global responses of the kidney to elevated aldosterone levels are determined by the coordinate action of different constituent tubule cells, including principal cells, intercalated cells and distal convoluted tubule cells. Recent studies on genetic mutations causing aldosterone overproduction have identified the molecules involved in aldosterone biosynthesis in the adrenal gland, and there is also increasing evidence for mechanisms and signaling pathways regulating the balance between renal NaCl reabsorption and K+ secretion, the two major effects of aldosterone. In particular, recent studies have demonstrated that mineralocorticoid receptor in intercalated cells is selectively regulated by phosphorylation, which prevents ligand binding and activation. Moreover, the ubiquitin ligase complex composed of Kelch-like 3 and Cullin 3 acts downstream of angiotensin II and plasma K+ alterations, regulating Na-Cl cotransporter independently of aldosterone in distal convoluted tubule cells. These and other effects are integrated to produce appropriate kidney responses in a high-aldosterone state, and are implicated in fluid and electrolyte disorders in humans. This review summarizes the current knowledge on mechanisms modulating mineralocorticoid receptor and its downstream effectors in the distal nephron.


Improvement in Protein Domain Identification Is Reached by Breaking Consensus, with the Agreement of Many Profiles and Domain Co-occurrence.
Traditional protein annotation methods describe known domains with probabilistic models representing consensus among homologous domain sequences. However, when relevant signals become too weak to be identified by a global consensus, attempts for annotation fail. Here we address the fundamental question of domain identification for highly divergent proteins. By using high performance computing, we demonstrate that the limits of state-of-the-art annotation methods can be bypassed. We design a new strategy based on the observation that many structural and functional protein constraints are not globally conserved through all species but might be locally conserved in separate clades. We propose a novel exploitation of the large amount of data available: 1. for each known protein domain, several probabilistic clade-centered models are constructed from a large and differentiated panel of homologous sequences, 2. a decision-making protocol combines outcomes obtained from multiple models, 3. a multi-criteria optimization algorithm finds the most likely protein architecture. The method is evaluated for domain and architecture prediction over several datasets and statistical testing hypotheses. Its performance is compared against HMMScan and HHblits, two widely used search methods based on sequence-profile and profile-profile comparison. Due to their closeness to actual protein sequences, clade-centered models are shown to be more specific and functionally predictive than the broadly used consensus models. Based on them, we improved annotation of Plasmodium falciparum protein sequences on a scale not previously possible. We successfully predict at least one domain for 72% of P. falciparum proteins against 63% achieved previously, corresponding to 30% of improvement over the total number of Pfam domain predictions on the whole genome. The method is applicable to any genome and opens new avenues to tackle evolutionary questions such as the reconstruction of ancient domain duplications, the reconstruction of the history of protein architectures, and the estimation of protein domain age. Website and software: http://www.lcqb.upmc.fr/CLADE.


Rapid diagnosis of Theileria annulata by recombinase polymerase amplification combined with a lateral flow strip (LF-RPA) in epidemic regions.
Rapid and accurate diagnosis of Theileria annulata infection contributes to the formulation of strategies to eradicate this parasite. A simple and efficient diagnostic tool, recombinase polymerase amplification (RPA) combined with a lateral flow (LF) strip, was used in detection of Theileria and compared to other methods that require expensive instruments and skilled personnel. Herein, we established and optimized an LF-RPA method to detect the cytochrome b gene of T. annulata mitochondrial DNA from experimentally infected and field-collected blood samples. This method has many unparalleled characteristics, including that it is rapid (clear detection in 5min at constant temperature), sensitive (the limitation of detection is at least 2pg genomic DNA), and specific (no cross-reaction with other piroplasms that infect cattle). The LF-RPA assay was evaluated via testing 17 field blood samples and comparing the results of that of a PCR, showing 100% agreement, which demonstrates the ability of the LF-RPA assay to detect T. annulata infections in small number of samples (n=17). Taken together, the results indicate that this method could be used as an ideal diagnostic tool for detecting T. annulata in endemic regions with limited to fewer and local resources and could also be a potential technique for the surveillance and control of blood protozoa.


Meta-Analyses of Japanese Encephalitis Virus Infection, Dissemination, and Transmission Rates in Vectors.
The objective of this work was to summarize and quantify Japanese encephalitis virus (JEV) infection, dissemination, and transmission rates in mosquitoes, using a meta-analysis approach. Data were obtained from experimental studies, gathered by means of a systematic review of the literature. Random-effects subgroup meta-analysis models by mosquito species were fitted to estimate pooled estimates and to calculate the variance between studies for three outcomes of interest: JEV infection, dissemination, and transmission rates in mosquitoes. To identify sources of heterogeneity among studies and to assess the association between different predictors (mosquito species, virus administration route, incubation period, and diagnostic method) with the outcome JEV infection rate in vectors, we fitted univariable meta-regression models. Mosquito species and administration route represented the main sources of heterogeneity associated with JEV infection rate in vectors. This study provided summary effect size estimates to be used as reference for other investigators when assessing transmission efficiency of vectors and explored sources of variability for JEV infection rates in vectors. Because transmission efficiency, as part of vector competence assessment, is an important parameter when studying the relative contribution of vectors to JEV transmission, our findings contribute to further our knowledge, potentially moving us toward more informed and targeted actions to prevent and control JEV in both affected and susceptible regions worldwide.


Discovery and functional analysis of lncRNAs: Methodologies to investigate an uncharacterized transcriptome.
It is known that more than 70% of mammalian genomes are transcribed, yet the vast majority of transcripts do not code for proteins. Are these noncoding transcripts merely transcriptional noise, or do they serve a biological purpose? Recent developments in genomic analysis technologies, especially sequencing methods, have allowed researchers to create a large atlas of transcriptomes, study subcellular localization, and investigate potential interactions with proteins for a growing number of transcripts. Here, we review the current methodologies available for discovering and investigating functions of long noncoding RNAs (lncRNAs), which require a wide variety of applications to study their potential biological roles. This article is part of a Special Issue entitled: Clues to long noncoding RNA taxonomy1, edited by Dr. Tetsuro Hirose and Dr. Shinichi Nakagawa.


Novel Outer Retinal Columnar Abnormalities (ORCA) and Non-Vasogenic Cystoid Macular Edema in Dense Deposit Disease.
To describe the occurrence of bilateral outer retinal columnar abnormalities, non-vasogenic cystoid macular edema, and drusen in the context of dense deposit disease. Case report. An 18-year-old female with dense deposit disease was referred to our specialist center for diagnosis and management with findings consistent with bilateral non-vasogenic cystoid macular edema and drusen. She was followed up in our clinic for forty months and treated with acetazolamide and ketorolac drops. Baseline examination revealed bilateral visual acuity (VA) reduction, and macular elevation with peripapillary drusen on fundus biomicroscopy. Optical coherence tomography revealed bilateral hyporeflective cystoid central macula changes, microcystoid changes with increased central subfield thickness (>450 microns), and outer retinal columnar abnormalities (ORCAs). Fluorescein angiography showed no evidence of macular leakage. Electrodiagnostic testing was within normal limits. Over the course of follow-up, she received treatment with acetazolamide 250mg BD PO and ketorolac 0.5% eye drops, with a partial reduction in her edema and improvement in VA. Dense deposit disease is a rare disease secondary to complement cascade dysregulation, associated with drusen. To the best of our knowledge, this is the first report of bilateral non-vasogenic cystoid macular edema and ORCA in a young female patient with dense deposit disease, confirmed with multimodal imaging.


Posttranslational modifications and secretion efficiency of immunogenic hepatitis B virus L protein deletion variants.
Subviral particles of hepatitis B virus (HBV) composed of L protein deletion variants with the 48 N-terminal amino acids of preS joined to the N-terminus of S protein (1-48preS/S) induced broadly neutralizing antibodies after immunization of mice with a Semliki Forest virus vector. A practical limitation for use as vaccine is the suboptimal secretion of such particles. The role of the N-terminal preS myristoylation in the cellular retention of full-length L protein is described controversially in the literature and the relation of these data to the truncated L protein was unknown. Thus, we studied the effect of preS myristoylation signal suppression on 1-48preS/S secretion efficiency, glycosylation and subcellular distribution. The findings are that 1-48preS/S is secreted, and that removal of the N-terminal myristoylation signal in its G2A variant reduced secretion slightly, but significantly. The glycosylation pattern of 1-48preS/S was not affected by the removal of the myristoylation signal (G2A mutant) but was different than natural L protein, whereby N4 of the preS and N3 of the S domain were ectopically glycosylated. This suggested cotranslational translocation of 1-48preS in contrast to natural L protein. The 1-48preS/S bearing a myristoylation signal was localized in a compact, perinuclear pattern with strong colocalization of preS and S epitopes, while the non-myristoylated mutants demonstrated a dispersed, granular cytoplasmic distribution with weaker colocalization. The large deletion in 1-48preS/S in presence of the myristoylation site facilitated formation and secretion of protein particles with neutralizing preS1 epitopes at their surface and could be a useful feature for future hepatitis B vaccines.


A toolbox for intensity modulated radiation therapy optimization.
We have designed a toolbox that provides an environment for testing radiotherapy optimization techniques, objective functions, and constraints. A set of three-dimensional (3D) pencil beam dose distributions have been computed for a cylindrical phantom. The 6 MV pencil beams were computed using a superposition-based dose engine commissioned for an Elekta SL20 linear accelerator. Due to the cylindrical symmetry of the phantom, the pencil beam dose distributions for any arbitrary beam angle can be determined by simply rotating the pencil beam data sets. Thus, the full accuracy is maintained without the need for additional dose calculations or large data storage requirements. In addition to the pencil beam data sets, tools are included for (1) rotating the pencil beams, (2) calculating the beam's eye view, (3) drawing structures, (4) writing the pencil beam dose data out to the optimizer, and (5) visualizing the optimized results. The pencil beam data sets and the corresponding tools are available for download at http://medschool.umaryland.edu/departments/radiationoncology/pencilbeam/. With this toolbox, researchers will have the ability to rapidly test new optimization techniques and formulations for intensity modulated radiation therapy and 3D conformal radiotherapy.


Clinical Practice Guidelines for Prevention, Diagnosis and Management of Early and Delayed-onset Ocular Injuries Due to Mustard Gas Exposure.
To develop clinical practice guidelines (CPGs) for prevention, diagnosis, treatment and follow-up of ocular injuries caused by exposure to mustard gas. The clinical questions were designed by the guideline team. Websites and databases including National Guidelines Clearinghouse, National Institute for Clinical Excellence, Cochrane, and PubMed were searched to find related CPGs and explore possible answers to the clinical questions. Since there were no relevant CPGs in the literature, related articles in Persian and English languages were extracted. Each article along with its level of evidence was summarized. Additionally, hand search was performed by looking the reference list of each article. Consequently, recommendations were developed considering the clinical benefits and side effects of each therapeutic modality. The recommendations were re-evaluated in terms of customization criteria. All recommendations along with the related evidence were scored from 1 to 9 by experts from all medical universities of Iran. The level of agreement among the experts was evaluated by analyzing the given scores. The agreement was achieved for all recommendations. The experts suggested a number of minor modifications which were applied to the recommendations. Finally, CPGs were developed with 98 recommendations under three major domains including prevention of injury, diagnosis and management of the acute and delayed-onset mustard gas ocular injuries. Considering the lack of CPGs for the prevention, diagnosis, and management of mustard gas-induced keratitis, these recommendations would be useful to prevent the serious ocular complications of mustard gas and standardize eye care services to the affected individuals.


The progression of bone and muscle atrophy in mice hind limb with immobilization.
This study investigated the time course of changes of bone and muscle atrophy in mice with immobilization by denervation and fixation. The animals were fifty-two male C57 BL/6J mice, aged 10 weeks old. Eight mice were used as the base line, and the remaining ones were cut at the sciatic nerve of the left hind limb and fixed with a plaster cast. At week 1, 2, 3, and 4 after the operation, a cross-sectional area of the rectus femoris muscles and bone mechanical strength with a three-point bending test of the femur and tibia were measured. The time course of changes of the bone mechanical strength and of the cross-sectional area of the rectus femoris muscles between the intact and experimental limbs in each period compared with the control limbs, was determined. The bone mechanical strength of the femur, tibia, and the cross-sectional area of the rectus femoris muscles of the experimental limbs significantly decreased compared with those of the intact limbs at week 4, 3, 2 and 1 after the operation (p<0.05). Compared with the intact limbs, the bone mechanical strength and the cross-sectional area of the rectus femoris muscles of the experimental limbs declined approximately 10% and 30%, respectively, during the experiment (p<0.05). It was demonstrated that bone and muscle atrophy occurred at an early stage after immobilization by denervation and fixation, and that both types of atrophy progressed simultaneously in the present study.


Isolation and long-term culture of primary ocular human immunodeficiency virus type 1 isolates in primary astrocytes.
Vitreous specimens from 14 HIV-1 infected persons undergoing medically indicated vitrectomy were assayed for the presence of infectious HIV-1 and viral tropism. Human primary fetal astrocytes, adult lymphocytes, or macrophages were exposed to vitreous in culture and and cells were then assayed for HIV-1 DNA by polymerase chain reaction amplification. We found that 11 of 14 patients tested carried ocular HIV-1 which replicated in one or more primary cell types; of the 13 vitreous samples tested in astrocytes, eight contained transmissible HIV-1. The three patients with no culturable ocular virus were in antiviral therapy at the time of vitrectomy. Comparison of envelope V3 sequences from astrocytes infected in culture to that in uncultured blood cells revealed 21% sequence divergence indicating that ocular HIV-1 transmitted to astrocytes was not recently derived from virus present in the blood. Two ocular samples transmissible to astrocytes were tested further and found capable of sustained replication by serial passage to uninfected astrocytes. However, the viral structural proteins produced by infected astrocytes were abnormal, p24 was absent and higher molecular weight Gag proteins were present. We conclude that the eye is a central nervous system compartment which frequently contains HIV-1 capable of replication in human astrocytes.


Medical-legal partnership: collaborating with lawyers to identify and address health disparities.
Medical-legal partnerships (MLPs) bring together medical professionals and lawyers to address social causes of health disparities, including access to adequate food, housing and income. Eighty-one MLPs offer legal services for patients whose basic needs are not being met. Besides providing legal help to patients and working on policy advocacy, MLPs educate residents (29 residency programs), health care providers (160 clinics and hospitals) and medical students (25 medical schools) about how social conditions affect health and screening for unmet basic needs, and how these needs can often be impacted by enforcing federal and state laws. These curricula include medical school courses, noon conferences, advocacy electives and CME courses. Four example programs are described in this paper. Established MLPs have changed knowledge (MLP | Boston-97% reported screening for two unmet needs), attitudes (Stanford reported reduced concern about making patients "nervous" with legal questions from 38% to 21%) and behavior (NY LegalHealth reported increasing resident referrals from 15% to 54%) after trainings. One developing MLP found doctors experienced difficulty addressing social issues (NJ LAMP-67% of residents felt uncomfortable). MLPs train residents, students and other health care providers to tackle socially caused health disparities.


Baltic sub-basin turnover times examined using the Rossby Centre Ocean model.
Not least when judging the possible effects of climate change it proves necessary to estimate the water-renewal rates of limited marine areas subject to pronounced external influences. In connection with the SWECLIM programme this has been undertaken for two ecologically sensitive sub-basins of the Baltic, viz. the Gulf of Riga and Gdansk Bay. For this purpose two methodologically different approaches have been employed, based on mass-balance budgets and analysis of Lagrangian trajectories, respectively. When compared to the results obtained using the Lagrangian technique, the box-model approach proved to be adequate for the Gulf of Riga representing a morphologically highly constrained basin, whereas it demonstrated certain shortcomings when applied to the more open topographic conditions characterizing Gdansk Bay.


[Molecular and immunological analyses suggest the absence of hydrophilic surface proteins in Leishmania (Viannia) panamensis].
The genus Leishmania is divided into two subgenera: Leishmania and Viannia. The two subgenera present several important differences such as the pathology they cause in susceptible hosts, their in vitro growth behavior, their genetic characteristics, and the expression pattern of several proteins, including those of the hydrophilic surface protein family. To characterize the hydrophilic surface protein family in Leishmania (Viannia) panamensis. The hasp genes were amplified in L. (V.) panamensis, using specific primers previously designed to amplify this gene in Leishmania (Leishmania) major. The PCR products were cloned, sequenced, and the sequences analyzed using common bioinformatics tools. Secondly, a serological screening was undertaken with an enzyme-linked immunosorbent assay and Western blot to detect specific antibodies against the hydrophilic surface recombinant protein from L. (L.) major. A copy of a pseudogene was amplified in L. (V.) panamensis which was 60% homologous with the L. (L.) major orthologous gene. Antibodies responded to the hydrophilic surface recombinant proteins only in sera from patients with visceral leishmaniasis [Leishmania (Leishmania) chagasi]. These results suggest the lack of a functional hasp gene in L. (V.) panamensis, suggesting probably the loss of the complete gene family in this species of the Viannia subgenus.


Effect of different muscle contraction interventions using an isokinetic dynamometer on muscle recovery following muscle injury.
The objective of the present study was to examine the effects of different muscle contraction interventions using isokinetic dynamometers on the muscle recovery after muscle injury caused by eccentric exercise. A total of 28 healthy male adults participated in the present study and each subject was randomly assigned to eccentric muscle contraction (EC, n=7), concentric muscle contraction (CC, n=7), both eccentric and concentric muscle contraction (BEC, n=7), and control (CON, n=7) groups after performing eccentric exercise. In all groups, except the control group, a different type of muscle contraction intervention was applied repeatedly for 24-96 hr after eccentric exercise, and maximum strength, muscle soreness, and creatine kinase (CK) levels were measured. The results showed a significant association between time and group, with respect to maximum strength (P<0.05), muscle soreness (P<0.001), and CK (P<0.001) after eccentric exercise. Moreover, BEC group showed faster muscle recovery than other groups. In conclusion, implementing active strategies such as performing BEC using an isokinetic dynamometer after eccentric exercise may be effective in facilitating muscle recovery.


Mouse models and human islet transplantation sites for intravital imaging.
Human islet transplantations into rodent models are an essential tool to aid in the development and testing of islet and cellular-based therapies for diabetes prevention and treatment. Through the ability to evaluate human islets in an in vivo setting, these studies allow for experimental approaches to answer questions surrounding normal and disease pathophysiology that cannot be answered using other in vitro and in vivo techniques alone. Intravital microscopy enables imaging of tissues in living organisms with dynamic temporal resolution and can be employed to measure biological processes in transplanted human islets revealing how experimental variables can influence engraftment, and transplant survival and function. A key consideration in experimental design for transplant imaging is the surgical placement site, which is guided by the presence of vasculature to aid in functional engraftment of the islets and promote their survival. Here, we review transplantation sites and mouse models used to study beta cell biology in vivo using intravital microscopy and we highlight fundamental observations made possible using this methodology.


Indications for removal of etonogestrel implant within two years of use in Jos, Nigeria.
Implanon is a new long-term and reversible sub-dermal contraceptive implant in Nigeria. It is a single rod containing 68mg of etonogestrel meant to offer contraception for three years and marketed by Organon. To determine the indications for removal of Implanon rods from clients within a two-year period. A retrospective review of 30 consecutive Implanon removals within the study period. The fertility regulation unit of the department of obstetrics and gynaecology of the Jos University Teaching Hospital, North-Central Nigeria. A total of 30 clients requested for and had their Implanon rods removed out of 669 insertions constituting 95.5% crude continuation rate in the second year. The clients were of mean age 31.4 +/- 6.2 years, mean parity 2.9 +/- 1.8 and mean number of living children 2.7 +/- 1.6. There was an average weight gain of 1.9 kg. The most common indication for removal was menstrual disruption (33.3%). Desire for another pregnancy closely followed (30.0%). Weight gain was another indication for discontinuation (13.3%). Two women were pregnant at insertion of the implant. There was one failure of the method with pregnancy as a result. Spousal disapproval was an indication for removal in two cases. Like all progestin-only contraceptive methods, menstrual disruption was the most common indication for removal of implants. Inadvertent insertion of implants with existing pregnancy is of concern and should be avoided as much as is possible. In doubtful cases at insertion, this insertion should be deferred or serum beta-HCG should be assessed to exclude chemical pregnancy.


Serum IGF-1 is insufficient to restore skeletal size in the total absence of the growth hormone receptor.
States of growth hormone (GH) resistance, such those observed in Laron dwarf patients, are characterized by mutations in the GH receptor (GHR), decreased serum and tissue IGF-1 levels, impaired glucose tolerance, and impaired skeletal acquisition. IGF-1 replacement therapy in such patients increases growth velocity but does not normalize growth. Herein we combined the GH-resistant (GHR knockout [GHRKO]) mouse model with mice expressing the hepatic Igf-1 transgene (HIT) to generate the GHRKO-HIT mouse model. In GHRKO-HIT mice, serum IGF-1 levels were restored via transgenic expression of Igf-1, allowing us to study how endocrine IGF-1 affects growth, metabolic homeostasis, and skeletal integrity. We show that in a GH-resistant state, normalization of serum IGF-1 improved body adiposity and restored glucose tolerance but was insufficient to support normal skeletal growth, resulting in an osteopenic skeletal phenotype. The inability of serum IGF-1 to restore skeletal integrity in the total absence of GHR likely resulted from reduced skeletal Igf-1 gene expression, blunted GH-mediated effects on the skeleton that are independent of serum or tissue IGF-1, and poor delivery of IGF-1 to the tissues. These findings are consistent with clinical data showing that IGF-I replacement therapy in patients with Laron syndrome does not achieve full skeletal growth.


MECP2 missense mutations outside the canonical MBD and TRD domains in males with intellectual disability.
Methyl-CpG-binding protein 2 (MeCP2) is a nuclear protein highly expressed in neurons that is involved in transcriptional modulation and chromatin remodeling. Mutations in MECP2 in females are associated with Rett syndrome, a neurological disorder characterized by a normal neonatal period, followed by the arrest of development and regression of acquired skills. Although it was initially thought that MECP2 pathogenic mutations in males were not compatible with life, starting from 1999 about 60 male patients have been identified and their phenotype varies from severe neonatal encephalopathy to mild intellectual disability. Targeted next-generation sequencing of a panel of intellectual disability related genes was performed on two unrelated male patients, and two missense variants in MECP2 were identified (p.Gly185Val and p.Arg167Trp). These variants lie outside the canonical methyl-CpG-binding domain and transcription repression domain domains, where the pathogenicity of missense variants is more difficult to establish. In both families, variants were found in all affected siblings and were inherited from the asymptomatic mother, showing skewed X-chromosome inactivation. We report here the first missense variant located in AT-hook domain 1 and we underline the importance of MECP2 substitutions outside the canonical MeCP2 domains in X-linked intellectual disability.


Disconnected lives: Women with intellectual disabilities in conflict with the law.
Women with intellectual/developmental disabilities in conflict with the law experience childhood trauma, substance abuse and intimate partner violence but continue to have difficulty accessing appropriate therapeutic services, both within correctional settings and upon discharge. The aim of this study is to explore women's service needs and to critically assess whether the available services are meeting their identified needs. Semi-structured interviews were conducted with 16 women with intellectual/developmental disabilities who were in the Special Needs Unit of a women's correctional centre from December 2014 to March 2015. Women with intellectual/developmental disabilities struggle to manage the impact of intergenerational trauma, exacerbated by issues of substance abuse and addiction, poor coping skills and minimal education, all of which impact their sense of well-being. This study highlights the need for increasing trauma treatment for women with intellectual/developmental disabilities, and emphasizes the need for accessible intervention to facilitate coping, trauma processing and community integration.


Cellular Redox Systems Impact the Aggregation of Cu,Zn Superoxide Dismutase Linked to Familial Amyotrophic Lateral Sclerosis.
Protein misfolding is implicated in neurodegenerative diseases such as ALS, where mutations of superoxide dismutase 1 (SOD1) account for about 20% of the inherited mutations. Human SOD1 (hSOD1) contains four cysteines, including Cys(57) and Cys(146), which have been linked to protein stability and folding via forming a disulfide bond, and Cys(6) and Cys(111) as free thiols. But the roles of the cellular oxidation-reduction (redox) environment in SOD1 folding and aggregation are not well understood. Here we explore the effects of cellular redox systems on the aggregation of hSOD1 proteins. We found that the known hSOD1 mutations G93A and A4V increased the capability of the thioredoxin and glutaredoxin systems to reduce hSOD1 compared with wild-type hSOD1. Treatment with inhibitors of these redox systems resulted in an increase of hSOD1 aggregates in the cytoplasm of cells transfected with mutants but not in cells transfected with wild-type hSOD1 or those containing a secondary C111G mutation. This aggregation may be coupled to changes in the redox state of the G93A and A4V mutants upon mild oxidative stress. These results strongly suggest that the thioredoxin and glutaredoxin systems are the key regulators for hSOD1 aggregation and may play critical roles in the pathogenesis of ALS.


Associations between polymorphisms in the vitamin D receptor and breast cancer risk.
Biological and epidemiologic data suggest that vitamin D levels may influence breast cancer development. The vitamin D receptor (VDR) is a crucial mediator for the cellular effects of vitamin D and additionally interacts with other cell-signaling pathways that influence cancer development. Because functional data exist on FOK1 and previous studies have suggested a relation between BSM1 and breast cancer risk, we evaluated the associations of the FOK1 and BSM1 VDR polymorphisms and breast cancer risk. In a case-control study nested within the Nurses' Health Study, we genotyped 1,234 incident cases (diagnosed between return of a blood sample in 1989-1990 and June 1, 2000) and 1,676 controls for FOK1, and 1,180 cases and 1,547 controls for BSM1. We observed a significantly increased risk of breast cancer among carriers of the ff genotype of FOK1 (multivariate odds ratio, 1.34; 95% confidence intervals, 1.06-1.69) compared with those with FF. We did not observe an association between polymorphisms in BSM1 and breast cancer risk (multivariate odds ratio, 0.93; 95% confidence intervals, 0.72-1.20) for BB versus bb). The FOK1 association did not vary significantly by menopausal status, estrogen, and progesterone receptor status of the tumors, or plasma levels of 25 hydroxyvitamin D or 1,25 dihydroxyvitamin D. Our results suggest that the VDR may be a mediator of breast cancer risk and could represent a target for cancer prevention efforts.


Rubik-Net: Learning Spatial Information via Rotation-Driven Convolutions for Brain Segmentation.
The accurate segmentation of brain tissue in Magnetic Resonance Image (MRI) slices is essential for assessing neurological conditions and brain diseases. However, it is challenging to segment MRI slices because of the low contrast between different brain tissues and the partial volume effect. 2-Dimensional (2-D) convolutional networks cannot handle such volumetric image data well because they overlook spatial information between MRI slices. Although 3-Dimensional (3-D) convolutions capture volumetric spatial information, they have not been fully exploited to enhance representative ability of deep networks; moreover, they may lead to overfitting with insufficient training data. In this paper, we propose a novel convolutional mechanism, termed Rubik convolution, to capture multi-dimensional information between MRI slices. Rubik convolution rotates the axis of a set of consecutive slices, enabling 2-D convolution kernels to extract features of each axial plane simultaneously. Next, feature maps are rotated back to fuse multidimensional information by the Max-View-Maps. Furthermore, we propose an efficient 2-D convolutional network, namely Rubik-Net, where the residual connections and the bottleneck structure are used to enhance information transmission and reduce the number of network parameters. The Rubik-Net shows promising results on iSeg2017, iSeg2019, IBSR and BrainWeb datasets in terms of segmentation accuracy. In particular, we achieved the best results in 95th percentile Hausdorff distance and average surface distance in cerebrospinal fluid segmentation on the most challenging iSeg2019 dataset. The experiments indicate that Rubik-Net improves the accuracy and efficiency of medical image segmentation. Moreover, Rubik convolution can be easily embedded into existing 2-D convolutional networks.


The COUP-TFII variant lacking a DNA-binding domain inhibits the activation of the Cyp7a1 promoter through physical interaction with COUP-TFII.
The COUP-TFII (chicken ovalbumin upstream promoter-transcription factor II) nuclear receptor, which is composed of a DNA-binding domain and a ligand-binding domain, exerts pleiotropic effects on development and cell differentiation by regulating the transcription of its target genes, including Cyp7a1 (cytochrome P450, family 7, subfamily a, polypeptide 1), which plays important roles in catabolism of cholesterol in the liver. Although multiple variants of COUP-TFII exist, their roles in the regulation of Cyp7a1 expression have not been elucidated. In the present study, we investigated the roles of COUP-TFII-V2 (variant 2), which lacks a DNA-binding domain, in the regulation of the transcriptional control of the Cyp7a1 gene by COUP-TFII in hepatocellular carcinoma cells. We found that COUP-TFII-V2 was significantly expressed in Huh7 cells, in which Cyp7a1 was not expressed. Furthermore, knockdown of COUP-TFII-V2 enhanced endogenous Cyp7a1 expression in Huh7 cells. Although COUP-TFII activates the Cyp7a1 promoter through direct binding to DNA, this activation was affected by COUP-TFII-V2, which physically interacted with COUP-TFII and inhibited its DNA-binding ability. Chromatin immunoprecipitation assays showed that COUP-TFII-V2 inhibited the binding of endogenous COUP-TFII to the intact Cyp7a1 promoter. The results of the present study suggest that COUP-TFII-V2 negatively regulates the function of COUP-TFII by inhibiting its binding to DNA to decrease Cyp7a1 expression.