Association of Haptoglobin Phenotype With Neurological and Cognitive Outcomes in Patients With Subarachnoid Hemorrhage.
To assess the association of haptoglobin (Hp) phenotype with neurological and cognitive outcomes in a large cohort of patients with subarachnoid hemorrhage (SAH). This prospective multicenter study enrolled patients with aneurysmal SAH between May 2015 and September 2020. The Hp phenotype was confirmed via Western blots. The relative intensities of α1 in individuals carrying Hp2-1 were compared with those of albumin. Multivariable logistic and Cox proportional-hazard regression analyses were used to identify the risk factors for 6-month and long-term outcomes, respectively. A total of 336 patients including the phenotypes Hp1-1 (n = 31, 9.2%), Hp2-1 (n = 126, 37.5%), and Hp2-2 (n = 179, 53.3%) were analyzed. The Hp phenotype was closely associated with 6-month outcome (p = 0.001) and cognitive function (p = 0.013), and long-term outcome (p = 0.002) and cognitive function (p < 0.001). Compared with Hp1-1 as the reference value, Hp2-2 significantly increased the risk of 6-month poor outcome (OR: 7.868, 95% CI: 1.764-35.093) and cognitive impairment (OR: 8.056, 95% CI: 1.020-63.616), and long-term poor outcome (HR: 5.802, 95% CI: 1.795-18.754) and cognitive impairment (HR: 7.434, 95% CI: 2.264-24.409). Long-term cognitive impairment based on the Hp phenotype was significantly higher in patients under 65 years of age (p < 0.001) and female gender (p < 0.001). A lower relative α1/albumin intensity (OR: 0.010, 95% CI: 0.000-0.522) was associated with poor outcome at 6 months but not cognitive impairment in patients with SAH expressing Hp2-1. Hp2-2 increased the risk of poor neurological outcomes and cognitive impairment compared with Hp1-1. For Hp2-1, higher relative α1 intensities were related to 6-month favorable outcomes.


A Special Approach for Stereotactic Radiofrequency Thermocoagulation of Hypothalamic Hamartomas With Bilateral Attachments to the Hypothalamus: The Transthird Ventricular Approach to the Contralateral Attachment.
Disconnection surgery for the treatment of epileptic hypothalamic hamartomas (HHs) is strategically difficult in cases with complex-shaped HHs, especially with bilateral hypothalamic attachments, despite its effectiveness. To evaluate the feasibility of a new approach for stereotactic radiofrequency thermocoagulation (SRT) using penetration of the third ventricle (SRT-TT) aiming to disconnect bilateral hypothalamic attachments in a single-staged, unilateral procedure. Ninety patients (median age at surgery, 5.0 years) who had HHs with bilateral hypothalamic attachments and were followed for at least 1 year after their last SRT were retrospectively reviewed. Thirty-three patients underwent SRT-TT as initial surgery. Of the 58 patients after mid-2013 when SRT-TT was introduced, 33 underwent SRT-TT and 12 (20.7%) required reoperation (ReSRT), whereas 20 of 57 patients (35.1%) without SRT-TT underwent reoperation. Reoperation was required in significantly fewer patients after mid-2013 (n = 12 of 58, 20.7%) than before mid-2013 (n = 15 of 32, 46.9%) ( P = .01). Final seizure freedoms were not different between before and after mid-2013 (gelastic seizure freedom, n = 30 [93.8%] vs n = 49 [84.5%] and other types of seizure freedom, n = 21 of 31 [67.7%] vs n = 32 of 38 [84.2%]). Persistent complications were less in SRT-TT than in ReSRT using the bilateral approach, but not significantly. However, hormonal replacement was required significantly more often in ReSRT using the bilateral approach (4 of 9, 44.4%) than in SRT-TT (3 of 32, 9.4%) ( P = .01). SRT-TT enabled disconnection of bilateral attachments of HHs in a single-staged procedure, which reduced the additional invasiveness of reoperation. Moreover, SRT-TT reduced damage to the contralateral hypothalamus, with fewer endocrinological complications than the bilateral approach.


Evolutionary conservation of biotinidase: implications for the enzyme's structure and subcellular localization.
Biotinidases from various species ranging from fungi and insects to human have specific amino acids, and regions that are evolutionarily conserved. These specific amino acids and regions are further supported by their homology to a variety of amidases and nitrilases and by the location of missense mutations that cause biotinidase deficiency in humans. Glu-Lys-Cys residues from three of these regions are considered the catalytic triad involved in the catalysis of the amide linkage. The last one-third of the biotinidase sequence is lacking in nitrilases-amidases, which do not bind biocytin or biotin, therefore, it is likely that the biocytin-biotin-binding site of biotinidase is within this portion of the molecule. Although there are many missense mutations at the far C-terminus of the enzyme, the function of this region is still unclear. Biotinidase may have different functions in different cells or in different subcellular compartments. Using computer programs that predict the subcellular localization of proteins based on their N-terminal signal peptides, microsomal localization resulting in secretion was predicted for biotinidase from all species, whereas there is little consistent support for mitochondrial or nuclear localization of the enzymes. Additional immunohistochemical studies of various human tissues at different stages of development are necessary to resolve the ambiguity of subcellular localization of biotinidase.


Radiotracer properties determined by high performance liquid chromatography: a potential tool for brain radiotracer discovery.
Previously, development of novel brain radiotracers has largely relied on simple screening tools. Improved selection methods at the early stages of radiotracer discovery and an increased understanding of the relationships between in vitro physicochemical and in vivo radiotracer properties are needed. We investigated if high performance liquid chromatography (HPLC) methodologies could provide criteria for lead candidate selection by comparing HPLC measurements with radiotracer properties in humans. Ten molecules, previously used as radiotracers in humans, were analysed to obtain the following measures: partition coefficient (Log P); permeability (P(m)); percentage of plasma protein binding (%PPB); and membrane partition coefficient (K(m)). Relationships between brain entry measurements (Log P, P(m) and %PPB) and in vivo brain percentage injected dose (%ID); and K(m) and specific binding in vivo (BP(ND)) were investigated. Log P values obtained using in silico packages and flask methods were compared with Log P values obtained using HPLC. The modelled associations with %ID were stronger for %PPB (r(2)=0.65) and P(m) (r(2)=0.77) than for Log P (r(2)=0.47) while 86% of BP(ND) variance was explained by K(m). Log P values were variable dependant on the methodology used. Log P should not be relied upon as a predictor of blood-brain barrier penetration during brain radiotracer discovery. HPLC measurements of permeability, %PPB and membrane interactions may be potentially useful in predicting in vivo performance and hence allow evaluation and ranking of compound libraries for the selection of lead radiotracer candidates at early stages of radiotracer discovery.


Synthesis, structure and theoretical study of mixed fluoro-trifluoromethyl derivatives of C60. Molecular structures of C60F18(CF3)6 and C60F16(CF3)6.
A series of novel mixed C60Fn(CF3)m compounds has been produced by trifluoromethylation of C60F18 with CF3I in ampoules at 380-420 degrees C. Two of these compounds, C60F18(CF3)6 and C60F16(CF3)6, have been characterized by X-ray crystallography, which has revealed addition of six CF3 groups to the C3v-C60F18 for the former and replacement/elimination of two outermost F atoms in the latter. Quantum chemical calculations have been employed to predict the most stable possible isomers of C60F16/18(CF3)6 in order to rationalize the experimental results.


Analysis of Readability, Quality, and Content of Online Information Available for "Stem Cell" Injections for Knee Osteoarthritis.
An increasing number of patients use the Internet to obtain health information, although online information is unregulated and highly variable. We aimed to assess the readability, quality, and content of online information available for "stem cell" injections for knee osteoarthritis. A cross-sectional analysis was performed on March 2019, inputting the search term "stem cells osteoarthritis" into the 3 most popular global search engines: Google, Bing, and Yahoo. The first 50 search results of each engine were evaluated/categorized. Readability was assessed using Flesch-Kincaid Ease/Grade Level. Quality/content was assessed through DISCERN score and a stem cell content score created for this study. Eighty-two websites were analyzed (18 academic websites, 21 commercial, 13 government/non-profit, 30 physician). Among all websites, mean Flesch-Kincaid readability was 35.9 with a grade level of 13.6. The average DISCERN score was 49.5/80 with statistically significant differences between academic vs physician websites (64.6 vs 38.1, P < .001), and commercial vs physician websites (52.3 vs 38.1, P = .001). Mean stem cell content score was 6.5/19 with a statistically significant difference between academic vs physician websites (8.5 vs 5.1, P = .007). Readability of online materials available for patients regarding "stem cell" treatment for knee osteoarthritis is significantly higher than the grade 6-8 recommended by the National Institutes of Health. The quality and content of websites is highly variable, with physician websites scoring especially low. Improving quality and readability of online materials that discuss risks/benefits of stem cell injections may potentially enhance the physician-patient therapeutic alliance and indirectly lead to better patient outcomes.


Role of ophthalmic nurses with visually impaired patients.
In 1987, it was estimated that over 13 million people in the world were visually impaired, and more than 27 to 35 million were classified as blind. Greater than 6 million visually impaired people live in the United States. There are many causes of visual impairment, including age-related macular degeneration, cataracts, glaucoma, diabetic retinopathy, autoimmune deficiency syndrome, and trauma. Some patients may be helped or visual loss delayed with surgery. Many may have dramatic visual improvement with optical devices, large-print publications, and improved lighting. The ophthalmic nurse may assist in accurately assessing the patient's vision and identifying his or her lifestyle and ability to function independently. Assisting the patient to overcome psychosocial and economic barriers and helping to obtain assistance is critical to enable patients to function independently. Ophthalmic nurses may be instrumental in increasing patients' awareness of community resources and services available to those who are visually impaired.


Recovery of partial 16S rDNA sequences suggests the presence of Crenarchaeota in the human digestive ecosystem.
Human feces collected from 10 healthy teenagers was analyzed for the presence of Crenarchaeota. After a first polymerase chain reaction (PCR) with Archaea-specific primers, a nested real-time PCR was performed using Crenarchaeota-specific primers. Real-time Crenarchaeotal PCR products detected from four subjects were cloned and the sequencing revealed that most of the partial 16S rRNA gene sequences were highly similar (> or = 97% homology) to sequences affiliated to the Sulfolobus group of the Crenarchaeota phylum. Our findings suggest for the first time that Crenarchaeota might be present in the microbiota of the human digestive ecosystem in which this phylum has never been found yet.


Vascular endothelial function is not related to serum uric acid in healthy adults.
Some experimental evidence suggests that uric acid impairs endothelial function. It is controversial if high uric acid levels and impaired endothelial function are related in healthy adults. In addition, the effect of uric acid on endothelial cells (ECs) of humans is unexplored. Data of 107 healthy adult volunteers were analyzed. The association between serum uric acid and endothelial-dependant dilation (EDD) and endothelial-independent dilation (EID) was evaluated by linear regression models. We also examined the relations between uric acid and systemic and cellular markers of inflammation and oxidative stress in all or subsets of participants. Uric acid levels and EDD were not related in unadjusted or adjusted models. There was a significant negative correlation between uric acid and EID in the pooled sample (r = -0.34, P = 0.005). This correlation remained significant after adjusting for demographics (P = 0.04) and was attenuated after adjusting for other cardiac risk factors (P = 0.12). Higher serum uric acid levels were found to correlate significantly with C-reactive protein (CRP) (r = 0.31, P = 0.002). Serum uric acid levels were not associated with brachial artery EC nuclear factor-κB (NF-κB) p65 or NADPH oxidase p47(phox) expression or with nitrotyrosine staining, but were inversely associated with EC manganese superoxide dismutase (MnSOD) expression (r = -0.5, P = 0.01, n = 25). Elevated serum uric acid is not associated with endothelial dysfunction among healthy adults, but is inversely related to EID and EC MnSOD, and positively related to systemic inflammation. These findings may have implications for cardiovascular risk in healthy adults.


Bone marrow macrophages induced to antigen presentation by lymphokines interact selectively with distinct T cells.
In vitro matured bone marrow-derived macrophages (BMM phi), which represent a pure population of M phi, were shown to act as antigen-presenting cells (APC) to the T cell clone ST2/K.9. This interaction was major histocompatibility complex restricted. Upon long-term culture in macrophage colony-stimulating factor, BMM phi were activated for antigen presentation by a 48-h pulse with lymphokine-containing supernatant of concanavalin A-stimulated rat spleen cells (Con A sup). The capacity of such activated M phi to function as APC decreased upon removal of Con A sup, and could be regenerated by a second pulse. This finding suggests that antigen presentation by mature M phi is a reversible function regulated by T cell factors. When the responsiveness of various T cell lines to antigen presented on BMM phi or spleen cells was compared, distinct activation requirements were observed for different T cells since lymphokine-activated BMM phi were not capable of inducing antigen-specific proliferation of all lines.


A preliminary evaluation of a new derived EEG index monitor in anesthetized patients.
An electroencephalogram (EEG) monitoring device, recently developed by Nicolet Biomedical, analyzes both high and low EEG frequencies. A processed derivative is obtained and displayed graphically and numerically on a monitor and may be used during anesthesia to indicate anesthetic "depth." However, radio-frequency interference from electrical equipment typically used in the operating room has the potential to interfere with the analysis of the high frequency components of the EEG. The objective of this study was to determine the ability of the Nicolet EEG monitoring device to function satisfactorily and effectively in the operating arena when used for anesthetized patients. A total of 40 patients undergoing surgery with general anesthesia were monitored using a prototype, processed EEG monitoring device. The device was used beginning prior to induction and continuing until emergence from anesthesia. Analysis was performed on 38 of the 40 patients. Brief interruption of the derived EEG Index occurred during use of the monopolar electrosurgical unit (ESU) as well as during stimulation of the facial nerve (used to monitor pharmacologic muscle relaxation). Derived EEG Index interruption was also noted during some movements of the patient's head or the EEG electrode wires. Changes in the derived EEG Index value accompanied changes in inhaled concentration of anesthetics and, in one patient, change in the derived EEG Index preceded patient movement that was not heralded by the typical cardiovascular indications of "light" anesthesia. The derived EEG Index monitoring system evaluated in this study functioned satisfactorily in the operating room setting in patients undergoing general anesthesia. Interference from ESU, facial nerve stimulation, and movement of the electrode wires caused brief interruptions of the derived EEG Index display and did not affect its utility in monitoring brain activity during anesthesia.


Increased Serum HMGB-1, ICAM-1 and Metalloproteinase-9 Levels in Buerger's Patients.
Thromboangiitis obliterans (TAO), or Buerger's disease, is an inflammatory occlusive disorder that affects the limb arteries of young smokers. In the aetiology of TAO the immune system appears to play a critical role; however, information on the aspects involved in the evolution of vascular tissue inflammation and of this disease are still limited. This study was carried out to investigate HMGB-1 (high mobility group box-1), MMP (matrix metalloproteinase)- 2, MMP-9, MMP-11 and ICAM (intercellular adhesion molecule)-1 circulating levels in subjects with Buerger's disease. Between January 2010 and December 2012, eight patients underwent surgical revascularization of the lower limbs and a specimen of the affected arterial wall was obtained for histological confirmation of Buerger's disease. A blood sample was collected on the same day for measuring HMGB-1, MMP-3, MMP-9 and ICAM-1 by western blot analysis. Controls (n=7) were healthy non-smokers. TAO subjects had a significant increase in HMGB-1, MMP-9 and ICAM-1 compared with controls (P<.0001), while no differences were observed in MMP-2 and MMP-11 levels. Histology confirmed a strong inflammatory infiltrate with signs of necrosis in the arterial wall. These data suggest a role for HMGB -1 in the vascular lesions associated with TAO, unveiling HMGB-1 as a potential target for treating this rare disease.


International health training. The Tulane experience.
At the heart of imminent healthcare reform are the future physicians of this decade. Medical educators will want to stimulate a desire within students to practice humanitarian, cost-effective, community-oriented primary care. Further studies to examine the effects of medical student experiences in international electives should be encouraged.


Latitudes mediate the association between influenza activity and meteorological factors: A nationwide modelling analysis in 45 Japanese prefectures from 2000 to 2018.
Cold and dry conditions were well-documented as a major determinant of influenza seasonality in temperate countries but the association may not be consistent when the climate in temperate areas is closer to that in sub-tropical areas. We hypothesized latitudes may mediate the association between influenza activity and meteorological factors in 45 Japanese prefectures. We used the weekly incidence of influenza-like illness of 45 prefectures from 2000 to 2018 as a proxy for influenza activity in Japan, a temperate country lying off the east coast of Asia. A combination of generalized additive model and distributed lag nonlinear model was adopted to investigate the associations between meteorological factors (average temperature, relative humidity, total rainfall, and actual vapour pressure, a proxy for absolute humidity) and the influenza incidence. Kendall's tau b (τ) and Spearman correlation coefficient (rs) between latitude and the adjusted relative risk (ARR) of each meteorological factor were also assessed. A higher vapour pressure was significantly associated with a lower influenza risk but the ARR strongly weakened along with a lower latitude (τ = -0.23, p-value = 0.02; rs = -0.33, p-value = 0.03). Lower temperature and lower relatively humidity were significantly associated with higher influenza risks in over 65% and around 40% of the prefectures respectively but the strength and significance of the correlations between their ARRs and latitude were weaker than that from vapour pressure. Even though the range of latitudes in Japan is small (26°N-43°N), the relationships between meteorological factors and influenza activity were mediated by the latitude. Our study echoed absolute humidity played a more important role in relating influenza risk, but we on the other hand showed its effect on influenza activity could be hampered in a low-latitude temperate region, which have a warmer climate. These findings thus offer a high-resolution characterization of the role of meteorological factors on influenza seasonality.


Injectable composites via functionalization of 1D nanoclays and biodegradable coupling with a polysaccharide hydrogel.
The use of injectable materials in minimally invasive surgical procedures could help in facing the bone diseases connected to the ageing of world population. To this aim, materials integrating the rheological properties of biocompatible polymers with the mechanical properties of 1D inorganic nanostructures represent promising scaffolds. Here we describe the preparation of hydrogel composites made of carboxymethyl cellulose (CMC) and halloysite nanotubes (HNT) as injectable materials for the local treatment of bone defects. The rheology and injectability of the materials reflects their structural properties, showing the possibility of successfully injecting the prepared composites over a large range of operative conditions.


Negative regulation of autophagy by UBA6-BIRC6-mediated ubiquitination of LC3.
Although the process of autophagy has been extensively studied, the mechanisms that regulate it remain insufficiently understood. To identify novel autophagy regulators, we performed a whole-genome CRISPR/Cas9 knockout screen in H4 human neuroglioma cells expressing endogenous LC3B tagged with a tandem of GFP and mCherry. Using this methodology, we identified the ubiquitin-activating enzyme UBA6 and the hybrid ubiquitin-conjugating enzyme/ubiquitin ligase BIRC6 as autophagy regulators. We found that these enzymes cooperate to monoubiquitinate LC3B, targeting it for proteasomal degradation. Knockout of UBA6 or BIRC6 increased autophagic flux under conditions of nutrient deprivation or protein synthesis inhibition. Moreover, UBA6 or BIRC6 depletion decreased the formation of aggresome-like induced structures in H4 cells, and α-synuclein aggregates in rat hippocampal neurons. These findings demonstrate that UBA6 and BIRC6 negatively regulate autophagy by limiting the availability of LC3B. Inhibition of UBA6/BIRC6 could be used to enhance autophagic clearance of protein aggregates in neurodegenerative disorders.


Effects of manipulating sedentary behavior on physical activity and food intake.
Sedentary behaviors have been correlated with obesity. We investigated whether changes in sedentary behaviors relate to changes in energy intake and/or physical activity. Experimental within-subject crossover design in which children participated in three 3-week phases: baseline and increased and decreased targeted sedentary behaviors. Thirteen 8- to 12-year-old, nonobese children. Sedentary behaviors were measured through the use of daily activity logs, physical activity measured with accelerometers, and energy intake measured by means of repeated 24-hour recalls collected during each phase. Energy intake, energy expenditure, and energy balance per day were calculated. Children showed significant (P <.001) increases of 50% and decreases of 53% in targeted sedentary behaviors from baseline during the increase and decrease phases, respectively. There was a significant (P =.05) increase in energy balance per day (+350.7 kcal) when sedentary behaviors were increased, as the result of an increase in energy intake per day (+250.9 kcal) and a decrease in energy expenditure (-99.8 kcal). No significant changes in energy balance were observed when sedentary behaviors were decreased. Increasing sedentary behaviors had a greater influence on physical activity and energy intake than reducing sedentary behavior in nonobese youth. In some children, changes in sedentary behaviors may be important to modify energy balance and prevent obesity.


The impact of diabetes education and peer support upon weight and glycemic control of elderly persons with noninsulin dependent diabetes mellitus (NIDDM).
We assessed diabetes education and peer support interventions as facilitators of weight loss and glycemic control in a community sample of 79 elderly persons with noninsulin-dependent diabetes mellitus (NIDDM). Different groups received: education only, education and peer support, and no treatment. Peer support was higher in groups where it was actively facilitated. Weight loss and reduction in level of glycemic control occurred within groups receiving both diabetes education and peer support.


Inferior vena caval involvement by adrenal cortical carcinoma.
A case of preoperative demonstration of inferior vena caval extension of tumor thrombus from adrenal cortical carcinoma is presented. Surgery showed that extension had occurred via the adrenal vein rather than by direct invasion. A search of the literature failed to uncover either any mention of preoperative attention to the possibility of caval involvement or of this particular mode of extension. Review of our experience with 17 adrenal cortical carcinomas suggested that carcinoma of the right adrenal gland involves the vena cava more commonly than does carcinoma of the left. We recommend that cavography be performed in the evaluation of adrenal carcinoma.


Prostaglandin receptors EP and FP are regulated by estradiol and progesterone in the uterus of ovariectomized rats.
Prostaglandins are important for female reproduction. Prostaglandin-E2 acts via four different receptor subtypes, EP1, EP2, EP3 and EP4 whereas prostaglandin-F2alpha acts through FP. The functions of prostaglandins depend on the expression of their receptors in different uterine cell types. Our aim was to investigate the expression of EPs and FP in rat uterus and to identify the regulation by estradiol, progesterone and estrogen receptor (ER) selective agonists. We performed four different rat experiments involving treatments with estradiol, progesterone and ER agonists. Real-time PCR and immunohistochemistry were employed to evaluate receptor expression. Our results showed that all mRNAs and proteins of EPs and FP are expressed in the rat uterus. The expression pattern and intensity of immunostaining vary between different cell types and treatments. The mRNA expression of all EPs and FP are downregulated by estradiol and the ERalpha specific agonist PPT, whereas the ERbeta specific agonist DPN downregulates only EP2 and EP4. The protein expression however, showed an increase in EP2 and EP3 after estradiol treatment. When treated with estradiol and progesterone in combination, the expressions of EP1 and EP3 are upregulated. Regulation of EPs and FP expression by estradiol appears to be mainly modulated via ERalpha for EP1, EP3 and FP, while EP2 and EP4 also are affected by the ERbeta selective ligand. Our immunohistochemical data shows a cell specific regulation of prostaglandin receptors under the influence of ovarian steroids, where EP2 is estrogen regulated in all uterine tissues examined. EP1 and EP3 are upregulated by the combination of estradiol and progesterone. Thus, our observations indicate that estradiol and progesterone regulate the mRNA and protein expression of EPs and FP in a receptor and tissue specific way.


Novel JAK1-selective benzimidazole inhibitors with enhanced membrane permeability.
The previously identified Janus kinase 1 (JAK1)-selective inhibitor, 1-(2-aminoethyl)-2-(piperidin-4-yl)-1H-benzo[d]imidazole-5-carboxamide (2), suffered from low cell permeability, which resulted in poor pharmacokinetic properties. In this study, by introducing less polar hydrogen bond donors at N(1) (a hydroxyalkyl or a methylaminoalkyl group) and C2 (a cyclohexanol group) positions, a series of novel benzimidazole derivatives were prepared, which exhibited selective JAK1 inhibitory activity (IC50 against JAK1=0.08-0.15μM; JAK1-selectivity=26-40 fold vs JAK2, 12-23 fold vs JAK3, and 38-54 fold vs Tyk2) along with significantly increased lipophilicity (3.3-15.8 times) as well as membrane permeability (6.3-12 times).


Crystal structure of a covalent intermediate of endogenous human arylsulfatase A.
The structures of human arylsulfatase A crystals soaked in solutions containing 4-methylumbelliferyl phosphate and O-phospho-DL-tyrosine have been determined at 2.7- and 3.2-A resolution, respectively. The formylglycine in position 69, a residue crucial for catalytic activity, was unambiguously identified in both structures as forming a covalent bond to the phosphate moiety. A hydroxyl group is present at the Cbeta of residue 69 and the formation of one out of two possible stereomeric forms is strongly favoured. The structures confirm the importance of the gem-diol intermediate in the arylsulfatase's catalytic mechanism. The presence of an apparently stable covalent bond is consistent with the weak phosphatase activity observed for human arylsulfatase A. The structures of the complexes suggest that phosphate ions and phosphate esters inhibit arylsulfatase in non-covalent and covalent modes, respectively. The metal ion present in the active site of arylsulfatase A isolated from human placenta is Ca(2+) and not Mg(2+) as was found in the structure of the recombinant enzyme.


Topical use of 5% acyclovir cream for the treatment of occult and verrucous equine sarcoids: a double-blinded placebo-controlled study.
Previous studies mention the use of topical acyclovir for the treatment of equine sarcoids. Success rates vary and since the bovine papillomavirus (BPV) lacks the presence of a kinase necessary to activate acyclovir, there is no proof of its activity against equine sarcoids. Twenty-four equine sarcoids were topically treated with acyclovir cream and 25 with a placebo. Both creams were applied twice daily during 6 months. Before the start of the treatment and further on a monthly basis, photographs and swabs were obtained. On the photographs, sarcoid diameter and surface area were measured and verrucosity of the tumours was quantified using a visual analog scale (VAS). The swabs were analysed by PCR for the presence of BPV DNA and positivity rates were calculated as the number of positive swabs divided by the total number of swabs for each treatment group at each time point. Success rates were not significantly different between both treatment groups. There was also no significant effect of treatment on sarcoid diameter, surface area or VAS score. For the swabs, a significantly higher BPV positivity rate was found for acyclovir treated tumours compared to placebo treated sarcoids only after 1 month of treatment and not at other time points. None of the results indicate that treatment with acyclovir yields any better results compared to placebo treatment.


Adaptive and Injury Response of Bone to Mechanical Loading.
Bone responds to supraphysiological mechanical loads by increasing bone formation. Depending on the applied strain magnitude (and other loading parameters) the response can be either adaptive (mostly lamellar bone) or injury (mostly woven bone). Seminal studies of Hert, Lanyon, and Rubin originally established the basic "rules" of bone mechanosensitivity. These were reinforced by subsequent studies using non-invasive rodent loading models, most notably by Turner et al. More recent work with these models have been able to explore the structural, transcriptional, and molecular mechanisms which distinguish the two responses (lamellar vs. woven). Wnt/Lrp signaling has emerged as a key mechanoresponsive pathway for lamellar bone. However, there is still much to study with regard to effects of ageing, osteocytes, other signaling pathways, and the molecular regulation that modulates lamellar vs. woven bone formation. This review summarizes not only the historical findings but also the current data for these topics.


Gene mutations and clinical prognosis of mucosal melanoma in different locations of head and neck.
Mucosal melanoma is the second most common subtype of melanoma in China and head and neck region is one of the main sites of this disease. Analyzed the phenotypes of C-Kit, NRAS, PDGFRA and BRAF genes in patients with in different locations to explore the characteristics of gene mutations. 96 patients were included in this study. C-Kit (exons 9, 11, 13, 17 and 18), NRAS (exons 1 and 2), PDGFRA (exons 12, 14 and 18) and BRAF (exons 11 and 15) were analyzed by PCR amplification and Sanger sequencing. 14 (14.58%) patients had C-Kit mutation, 6 (6.25%) had BRAF mutation, 23 (23.96%) had PDGFRA mutation, and 12 (12.50%) had NRAS mutation. The NRAS mutation (p = 0.037, 95%CI: 1.050-4.572) was an independent factor affecting distant metastasis and was most commonly found in the nasal cavity/paranasal sinuses (p = .043) while the BRAF mutation was more common in locations other than the nasal cavity/paranasal sinus (p = .008) and was associated with local recurrence. Gene phenotypes of mucosal melanoma in different locations has differences. Lesions in the nasal cavity and paranasal sinus should be assessed separately from other parts such as the nasopharynx.


The analyst's caseload as a family: transferences to fellow patients.
This article draws attention to a topic that has been mostly ignored in our literature: the manifold transference meanings of our patients' reactions to our other patients. Central aspects of the patient's transference may be displaced onto the analyst's other patients. The author includes clinical examples of hospitalized patients, where such transferences to other patients are often more dramatic and obvious, as well as examples from neurotic patients, where such transferences may be less dramatic and may tend to resolve more readily through interpretation. The displacement of transference feelings onto the analyst's other patients also has important implications for training analyses, where analysands are likely to have multiple relationships with the analyst's other analysands, supervisees, and students.


The PD-1/PD-L1 inhibitory pathway is altered in pre-eclampsia and regulates T cell responses in pre-eclamptic rats.
The programmed cell death-1(PD-1)/PD-ligand 1 (PD-L1) pathway is critical to immune homeostasis by promoting regulatory T (Treg) development and inhibiting effector T (such as Th17) cell responses. However, the association between the PD-1/PD-L1 pathway and the Treg/Th17 imbalance has not been fully investigated in pre-eclampsia (PE). In this study, we observed an inverse correlation between the percentages of Treg and Th17 cells, and the expression of PD-1 and PD-L1 on the two subsets also changed in PE compared with normal pregnancy. We further explored their relationship in vivo using the L-NG-Nitroarginine Methyl Ester (L-NAME) induced PE-like rat models, also characterized by Treg/Th17 imbalance. Administration of PD-L1-Fc protein provides a protective effects on the pre-eclamptic models, both to the mother and the fetuses, by reversing Treg/Th17 imbalance through inhibiting PI3K/AKT/m-TOR signaling and enhancing PTEN expression. In addition, we also observed a protective effect of PD-L1-Fc on the placenta by reversing placental damages. These results suggested that altered PD-1/PD-L1 pathway contributed to Treg/Th17 imbalance in PE. Treatment with PD-L1-Fc posed protective effects on pre-eclamptic models, indicating that the use of PD-L1-Fc might be a potential therapeutic target in PE treatment.


Aberrant brain functional and structural developments in MECP2 duplication rats.
Transgenic animal models with homologous etiology provide a promising way to pursue the neurobiological substrates of the behavioral deficits in autism spectrum disorder (ASD). Gain-of-function mutations of MECP2 cause MECP2 duplication syndrome, a severe neurological disorder with core symptoms of ASD. However, abnormal brain developments underlying the autistic-like behavioral deficits of MECP2 duplication syndrome are rarely investigated. To this end, a human MECP2 duplication (MECP2-DP) rat model was created by the bacterial artificial chromosome transgenic method. Functional and structural magnetic resonance imaging (MRI) with high-field were performed on 16 male MECP2-DP rats and 15 male wildtype rats at postnatal 28 days, 42 days, and 56 days old. Multimodal fusion analyses guided by locomotor-relevant metrics and social novelty time separately were applied to identify abnormal brain networks associated with diverse behavioral deficits induced by MECP2 duplication. Aberrant functional developments of a core network primarily composed of the dorsal medial prefrontal cortex (dmPFC) and retrosplenial cortex (RSP) were detected to associate with diverse behavioral phenotypes in MECP2-DP rats. Altered developments of gray matter volume were detected in the hippocampus and thalamus. We conclude that gain-of-function mutations of MECP2 induce aberrant functional activities in the default-mode-like network and aberrant volumetric changes in the brain, resulting in autistic-like behavioral deficits. Our results gain critical insights into the biomarker of MECP2 duplication syndrome and the neurobiological underpinnings of the behavioral deficits in ASD.


Large-scale Meta-analysis Suggests Low Regional Modularity in Lateral Frontal Cortex.
Extensive fMRI study of human lateral frontal cortex (LFC) has yet to yield a consensus mapping between discrete anatomy and psychological states, partly due to the difficulty of inferring mental states from brain activity. Despite this, there have been few large-scale efforts to map the full range of psychological states across the entirety of LFC. Here, we used a data-driven approach to generate a comprehensive functional-anatomical mapping of LFC from 11 406 neuroimaging studies. We identified putatively separable LFC regions on the basis of whole-brain co-activation, revealing 14 clusters organized into 3 whole-brain networks. Next, we generated functional preference profiles by using multivariate classification to identify the psychological states that best predicted activity within each cluster. We observed large functional differences between networks, suggesting brain networks support distinct modes of processing. Within each network, however, we observed relatively low functional specificity, suggesting discrete psychological states are not strongly localized to individual regions; instead, our results are consistent with the view that individual LFC regions work as part of distributed networks to give rise to flexible behavior. Collectively, our results provide a comprehensive synthesis of a diverse neuroimaging literature using relatively unbiased data-driven methods.


Severity of Hyperemesis Gravidarum and Associated Maternal factors.
Hyperemesis gravidarum is the most severe form of nausea and vomiting in pregnancy. It is one of the most common cause of early pregnancy admissions and associated with various maternal risk factors . Very few studies have been conducted among Nepalese women with hyperemesis gravidarum. This study aims to identify various maternal risk factors among Nepalese women and its severity using pregnancy unique quantification of emesis scoring. This is a cross sectional observational study conducted at Paropakar Maternity and Women's Hospital. Total duration of the study period was for one year from February 2018 to Janurary 2019. A total of 144 patients meeting the inclusion criteria were included in the study.The severity of the hyperemesis gravidarum was assessed and classified using pregnancy unique quantification of emesis scoring. Various maternal demographic, obstetric and personal factors were studied in relation to the incidence and severity of hyperemesis gravidarum. Moderate (49.30%) to severe(50.69%) hyperemesis gravidarum were admitted in the hospital. Most women were nulliparous from 20-24 years age group. Among all categories of BMI, underweight had more severe hyperemesis gravidarum (63.63%) and overweight patient had increased incidence of moderate hyperemesis gravidarum (66.66%). Women with previous dysmenorrhea had severe hyperemesis gravidarum (54.05%) and non-smoker had severe hyperemesis gravidarum (52.03%) while smoker had moderate hyperemesis gravidarum (57.14%). Pregnant women of age group of 20-24 years, nulliparity and underweight were associated with severe hyperemesis gravidarum.


Can Functionally Mature Islet β-Cells be Derived from Pluripotent Stem Cells? A Step Towards Ready-To-Use β-Cells in Type 1 Diabetes.
Pluripotent Stem Cells [PSCs] are emerging as an excellent cellular source for the treatment of many degenerative diseases such as diabetes, ischemic heart failure, Alzheimer's disease, etc. PSCderived pancreatic islet β-cells appear to be a promising therapy for type 1 diabetic patients with impaired β-cell function. Several protocols have been developed to derive β-cells from PSCs. However, these protocols produce β-like cells that show low glucose stimulated insulin secretion (GSIS) function and mirror GSIS profile of functionally immature neonatal β-cells. Several studies have documented a positive correlation between the sirtuins (a family of ageing-related proteins) and the GSIS function of adult β-cells. We are of the view that the GSIS function of PSC-derived β-like cells could be enhanced by improving the function of sirtuins in them. Studying the sirtuin expression and activation pattern during the β-cell development and inclusion of the sirtuin activators and inhibitor cocktail (specific to a developmental stage) in the present protocols may help us derive functionally mature, ready-to-use β- cells in-vitro making them suitable for transplantation in type 1 diabetes.


Transepidermal water loss in infants born at 24 and 25 weeks of gestation.
The rate of evaporation of water from the skin of 13 infants born at 24 (n=3) and 25 (n=10) weeks of gestation was measured on the first day after birth and at postnatal ages of 1, 3, 7 and 28 d, using the gradient method. Transepidermal water loss was estimated from this rate and corrected to an ambient relative humidity (RH) of 50%. Transepidermal water loss, corrected to 50% RH, was high on the first day after birth (58.4+/-14.8 g m(-2) h(-1)) and remained at the same level during the second day (59.3+/-17.6 g m(-2)h(-1)). It then decreased significantly to 43.8+/-9.5 at a postnatal age of 3 d, 36.1+/-12.6 at 7 d and 24.2+/-7.7 g m(-2) h(-1) at 28 d (p < 0.001). Within the group investigated, there was no significant correlation between transepidermal water loss and body or skin temperature, birth weight, gender, mode of delivery or gestational age. Transepidermal water loss on the first day after birth was somewhat lower than the highest losses previously found in infants born at 25 weeks of gestation, and of the same magnitude as previously reported for infants born at 25-27 weeks. Transepidermal water loss at postnatal ages of 1, 3, 7 and 28 d in the present study was higher than that previously found in the group of infants born at 25-27 weeks. In conclusion, in infants born at 24-25 completed weeks of gestation transepidermal water loss was high immediately after birth and decreased with increasing postnatal age, but at a slower rate than previously reported for slightly more mature infants.


Thymoindependent antigentic stimulation in nude mice: response to polyvinylpyrrolidone and adjuvant effects of Corynebacterium parvum and LH1.
The humoral thymoindependent response to polyvinylpyrrolidone (PVP) measured by the Farr test, was not different in athymic nude and phenotypically normal mice. In athymic and normal mice, this response was significantly increased by Cornebacterium parvum administered at the time of immunization. A recently synthetized compound, LH1 (3alpha-N-[2 aminoethanol]-amino 5-alpha-pregnane-20-one) was, however, a better immunostimulant of the anti-PVP response in athymic mice.


Addiction and the law: confidentiality to disclosure.
With growing demands for access to medical records, confidentiality and privacy will increasingly compete with professional desire for collaboration and the need for reimbursement. Physicians need to be aware of medicolegal safeguards.


Drug supply for psoriasis - results from a national pharmacy network.
In Germany, drugs are to a large extent provided by pharmacies. Thus, investigations in pharmacies permit drug usage studies both on patients receiving prescribable drugs and using self-medication. The current study evaluated the quality of medical care, disease burden and spectrum of treatments for patients with psoriasis in a nationwide network of pharmacies. A nationwide cross-sectional study was conducted in 61 pharmacies. Patients with psoriasis vulgaris who came to the pharmacy to obtain antipsoriatic drugs or basic ointments were consecutively recruited, interviewed and asked to complete a standardized questionnaire. The questionnaire focused on socio-demographic characteristics and prior therapies for psoriasis. Furthermore, data on the patient's treatment satisfaction, disease-related burden, and treatment adherence were evaluated. In addition, the proportion and significance of health care providers for psoriasis as well as the number of patients using self-medication were assessed. The data on 241 patients show a high and long-lasting disease-related burden. A high utilization of resources was found. Dermatologists were the most frequently consulted providers (reported by 77.1 % of patients), followed by general practitioners (10.4 %). 3.5 % of patients were using self-medication. Self-reported adherence with treatment was moderate (71.6 %). Patient satisfaction varied considerably and demonstrated the need for improvement. Psoriasis is a socio-economically relevant disease. Health care is provided primarily by dermatologists. Surveying patients in a national network of pharmacies is a unique and effective way of collecting relevant "real world" data. Selection biases related to the health care setting are minimized.


Epidemiology of melanoma: is it still epidemic? What is the role of the sun, sunbeds, Vit D, betablocks, and others?
The incidence of cutaneous malignant melanoma is continuously increasing worldwide, but only minimal changes in mortality have been observed. This phenomenon has brought into question whether this increased incidence reflects a true or apparent melanoma epidemic. The most recent data suggest that this epidemiological trend may be explained by the existence of a certain degree of melanoma overdiagnosis, especially of thin lesions, which probably would never progress to advanced disease if left untreated. However, acute sun exposure and widespread use of sunbeds may also justify the increase in melanoma incidence. Recently, both vitamin D and beta-blocker use seem to play a beneficial role in melanoma progression.


Respiratory disease and suicide among U.S. coal miners: is there a relationship?
A case-control study was performed to test whether or not respiratory disease in coal miners presented a risk for suicide. While coal miners in general do not experience elevated rates of suicide, coal miners with respiratory disease have been found to have high rates of psychiatric disability, especially depressive reactions. Further, depression has been related to suicide. To test the hypothesis, 50 suicide deaths from four National Institute for Occupational Safety and Health cohorts of coal miners were matched by age at death to two series of controls, a noncancer, nonaccident control series, and a cancer control series. Using odds ratios (tested by chi-square) the risks of obstructive lung disease and coal workers pneumoconiosis were evaluated together with the risks of years of underground mining, cigarette smoking at the time of cohort creation, and ever having smoked cigarettes. Neither respiratory disease was found to pose a statistically elevated risk of suicide in this sample of U.S. white male coal miners.


The modified Bosworth technique for the treatment of acute traumatic Achilles tendon rupture.
Between 1983 and 1994, 15 patients (range 18 to 62 years) with acute traumatic Achilles tendon rupture, were treated surgically in our Department. We employed a modified Bosworth technique. The modifications were the use of a shorter strip of tendon and more secure fixation of the proximal and distal stump, than the original Bosworth technique. Postoperatively an above - knee plaster cast was applied with the knee flexed 30°-40° and the foot in a relaxed equinus position. The plaster cast was changed to a below - knee after 4 weeks and the foot gradually dorsiflexed to a neutral position until the 8th week, and then the plaster cast was removed. No patient had wound separation, infection or skin sloughs. After an average follow-up of 9 years, no rerupture has been reported and all the patients have returned to their pre injury activities.


Non-obstetric female genital tract trauma in Port Moresby, Papua New Guinea.
In a 3-year period there were 25 admissions to Port Moresby General Hospital because of non obstetric injuries to the female genital tract. The injuries were caused by voluntary coitus, rape, assault and falls. The most frequent injury was a laceration to the posterior fornix of the vagina. Although rape was frequently committed in Port Moresby, it was not the major cause of genital tract injury.


A prospective randomized study of the difference in diagnostic yield between endoscopic ultrasound-guided fine-needle aspiration (EUSFNA) needles with and without a side port in pancreatic masses.
Two 22G needles with similar designs, apart from the absence (A) or presence of a side port (B), are available for endoscopic ultrasound (EUS)-guided fine-needle aspiration (FNA). The side port was designed to increase diagnostic yield but this advantage was unproven. This study evaluated the difference in diagnostic yield between both needles in pancreatic masses. This was a prospective multicenter randomized cross-over study. Patients with pancreatic masses were randomized to one needle for the first two passes, followed by the other for the next two passes. A pathologist blinded to the needle assessed each puncture for cellularity and morphology. The diagnostic yield between both needles was compared. In total, 30 patients were recruited (mean lesion size: 3.5 cm, range: 1.2 - 6.3). Comparison of cellularity adequacy: first pass: A vs. B: 26/30 vs. 24/30 (P = 0.488): 2nd pass: A vs. B: 25/30 vs. 26/30 (P = 0.718). Comparison of diagnostic accuracy: first pass: A vs. B: 22/30 vs. 23/30 (P = 0.766); after two passes: A vs. B: 26/30 vs. 26/30 (P = 1.0). When all four passes were assessed, adequate cellularity was obtained in 29/30 and the correct diagnosis was obtained in 28/30 patients. There were no procedural complications. There was no significant difference in diagnostic yield between EUSFNA needles with or without a side port for pancreatic masses. NCT02092519.


Topoisomerase 3b is dispensable for replication of a positive-sense RNA virus--murine coronavirus.
A recent study demonstrated that a DNA-RNA dual-activity topoisomerase complex, TOP3B-TDRD3, is required for normal replication of positive-sense RNA viruses, including several human flaviviruses and coronaviruses; and the authors proposed that TOP3B is a target of antiviral drugs. Here we examined this hypothesis by investigating whether inactivation of Top3b can inhibit the replication of a mouse coronavirus, MHV, using cell lines and mice that are inactivated of Top3b or Tdrd3. We found that Top3b-KO or Tdrd3-KO cell lines generated by different CRISPR-CAS9 guide RNAs have variable effects on MHV replication. In addition, we did not find significant changes of MHV replication in brains or lungs in Top3B-KO mice. Moreover, immunostaining showed that Top3b proteins are not co-localized with MHV replication complexes but rather, localized in stress granules in the MHV-infected cells. Our results suggest that Top3b does not have a universal role in promoting replication of positive-sense RNA virus, and cautions should be taken when targeting it to develop anti-viral drugs.


Mitochondrial genome evidence suggests Cooperia sp. from China may represent a distinct species from Cooperia oncophora from Australia.
Cooperia spp. are parasitic nematodes parasitizing in small intestine of ruminants with a worldwide distribution. Infection of ruminants with Cooperia species can cause severe enteritis, causing significant socio-economic losses to the livestock industry. However, it is yet to know whether there is genetic diversity in mitochondrial (mt) DNA sequences of Cooperia nematodes from different geographic regions. The objective of the present study was to examine sequence difference in mt genomes between Cooperia sp. from China and other Cooperia species. We determined the sequences of the internal transcribed spacer (ITS-1 and ITS-2) of nuclear ribosomal DNA (rDNA) of 11 Cooperia specimens collected from the small intestine of a Tianzhu White yak in Gansu Province, northwestern China, which had 99% similarity with that of C. oncophora from Brazil (GenBank accession Number: AJ544290) in ITS-1, and 99% similarity with those from Denmark (AB245040), Scotland and Australia (AJ000032) in ITS-2, indicating that specimens used in the present study should at least represent parasites in Cooperia. We then determined the complete mt genome sequences of one representative specimen of Cooperia sp. from China (CspC), compared the mt DNA sequences with that of C. oncophora from Australia (COA, GQ888713), and conducted phylogenetic analysis with selected nematodes using both maximum likelihood (ML) and Bayesian inference (BI) methods based on both concatenated 12 PCGs, rrnL and rrnS sequences and partial cox2 sequences. The complete mt genome sequence of CspC (KY769271) is 13, 583 bp in length, which is 91 bp shorter than that from COA. The sequence difference over the entire mt genome between CspC and COA was 12.2% in nucleotide and 6.3% in inferred amino acids, with nad4L and nad1 being the most variable and the most conserved PCGs, respectively. Phylogenetic analysis indicated that CspC and COA were closely-related but distinct taxa. The determination of mt genome sequences for Cooperia sp. from China also provides novel resources for further studies of taxonomy, systematics and population genetics of Cooperia from different geographical locations.


Linarite and connellite dataset from Calabria region (Southern Italy): First evidence.
This dataset article contains mineralogical and chemical data of linarite and connellite sulfate minerals. These minerals were found included in the oxidized zones of cupriferous sulfide ores in the Fiumarella Mine, in Calabria region (Southern Italy). Linarite is a basic sulfate of copper and lead with the formula of PbCuSO4(OH)2 while connellite is a hydrated sulfate of copper with an ideal formula of Cu36Cl6(SO4)2(OH)62∙12H2O. Recently, in the mine of Fiumarella, in addition to primary minerals such as barite, galena, cerussite, anglesite, fluorite and chalcopyrite, wulfenite (PbMoO4) was also detected. Linarite consists of a prismatic bright blue crystal with a vitreous luster of micrometric size implanted upon on a matrix made up of barite. Connellite includes micrometric acicular tuft crystals protruding from matrix. Methods for obtaining the datasets include optical microscopy, micro X-ray Fluorescence and micro-Raman spectroscopy.


Subfascial endoscopic perforator vein surgery (SEPS) using the ultrasonic scalpel.
Subfascial endoscopic perforator vein surgery (SEPS) was recently introduced as a minimally invasive method to ligate incompetent perforating veins in patients with severe chronic venous insufficiency of the lower extremities. Herein we describe a technique in which we used a 5-mm ultrasonic scalpel for the transection of perforating veins in 16 SEPS performed in 14 patients. The use of the ultrasonic scalpel allowed for the precise coagulation and transection of the perforator vein with hemostasis, while avoiding the use of metal clips. Our initial results showed that the technique was feasible with minimal morbidity. We recommend the use of the ultrasonic scalpel as an alternative tool to transect perforating veins in SEPS.


Heteroduplex analysis of the sequence relationships between the genomes of Kirsten and Harvey sarcoma viruses, their respective parental murine leukemia viruses, and the rat endogenous 30S RNA.
The sequence relations between Kirsten murine sarcoma virus (Ki-SV), Harvey murine sarcoma virus (Ha-SV), and a rat endogenous 30S RNA were studied by electron microscope heteroduplex analysis. The sequence relationships between the sarcoma viruses and their respective parental murine leukemia viruses (Kirsten and Moloney murine leukemia viruses), as well as between the two murine leukemia viruses, were also studied. The only observed nonhomology feature of the Kirsten murine leukemia virus/Moloney murine leukemia virus heteroduplexes was a substitution loop with two arms of equal length extending from 1.80 +/- 0.18 kilobases (kb) to 2.65 +/- 0.27 kb from the 3' end of the RNA. It is believed that this feature lies in the env gene region of the viral genomes. The Ha-SV and Moloney murine leukemia virus genomes (respective lengths, 6.0 and 9.0 kb) were homologous in a 1.0 +/- 0.05-kb region at the 3' end and possibly over a 200-nucleotide region at the 5' ends; otherwise, they were nonhomologous. Ha-SV and Ki-SV (length, 7.5 kb) were homologous in the first 4.36 +/- 0.37-kb region from the 3' end and in a 0.70 +/- 0.15-kb region at the 5' end. In between, there was a nonhomology region, possibly containing a short (0.23-kb) region of partial or total homology. The heteroduplex analysis between rat endogenous 30S RNA and Ki-SV shows that there are mixed regions of sequence homology and nonhomology at both the 5' and 3' ends. However, there is a large (4-kb) region of homology between Ki-SV and the rat 30S RNA in the center of the genomes, with only a small nonhomology hairpin feature. These studies help to define the regions of homology between the Ha-SV and Ki-SV genomes with each other and with the rat endogenous 30S RNA. These regions may be related to the sarcoma genicity of the viruses. In particular, the 0.7-kb region of homology of Ha-SV with Ki-SV at the 5' ends may be related to the formation of a 21,000-dalton phosphoprotein in cells transformed by either virus.


Recent Advances in Celiac Disease.
Recent diagnostic advances have demonstrated that celiac disease is relatively common although most patients have less florid symptoms than previously recognised. The mucosal lesion of this autoimmune disorder depends on both adaptive and innate immune responses. The characteristic antibodies to tissue transglutaminase-2 (tTG-2) and deamidated gliadin peptides may be produced in persons possessing the relevant HLA-DQ genotypes if intact gliadin peptides can penetrate the epithelial barrier to reach antigen presenting cells. Progression from celiac autoimmunity to overt disease may depend on innate immune mechanisms, not HLA-restricted, where IL-15 is generated within the epithelial compartment. A specific innate immune response previously thought restricted to invertebrates, the encapsulation reaction, may contribute to mucosal volume expansion through recruitment of syndecan-expressing leukocytes and stimulated matrix production. It is notable that tissue transglutaminase is critical in this reaction in insects, and that the very few insects that can predate wheat, possess specific salivary or intestinal enzymes that degrade gluten. Animal models in HLA-DQ transgenic mice suggest that the microbial flora of the intestine may play a role in host responses and modulate the evolution of the disease. This suggests that therapeutic modulation of the microbiome may contribute to management of celiac disease. In developing world countries, there is a potential difficulty in histological diagnosis because of the widespread incidence of environmental enteropathy amongst apparently healthy children. Thus, recognition of local patterns of enteropathy will be important for histopathologists, and high titre tTG-2 autoantibody titres may hold considerable diagnostic significance.


Differences in wasting assessment through Middle-Upper Arm Circumference (MUAC) adjusted by sex, age and geographic origin for children aged 6-59 months: New reference based on anthropometric surveys from 22 low-and-middle-income countries.
The Middle Upper Arm Circumference (MUAC) bracelet is a widely used instrument in public health assessments and humanitarian assistance projects. The WHO guidelines present a universal cut-off point of 115 mm to determine whether a child has severe acute malnutrition. The objective of this study is to analyze the existing differences in the MUAC for boys and girls aged between 6 and 59 months, from 22 countries distributed in three different continents, in contrast to the use of this single cut-off point. In addition, the creation of MUAC growth charts is presented for reference use. This study was carried out with a database developed by Action Against Hunger, composed, after the data pre-processing phase, of 97 921 individuals without anthropometric failure from African, Asian, and American continents. MUAC measurements were compared between countries, dividing by sex and age groups. A k-means method was used to create country clusters to allow comparisons and the variability was resumed using a Principal Component Analysis. For each cluster, growth curves were created and smoothed using the LOESS method. Our research has revealed the existence of differences in the MUAC between countries in both, males and females, although with different trends. The evidence was confirmed with the creation of two clusters using the k-means method, which, when graphically represented by the Principal Component Analysis, showed that the MUAC was clearly different. There were also differences between males and females within each cluster, where growth curves did not overlap in any age group. All statistical analysis indicate that there are differences in the MUAC values for children without anthropometric failure between countries, but also between sexes. With this research, a new reference is proposed that consider the existing variability between human populations to improve the precision in the determination of severe acute malnutrition in children.


Amperometry in Single Cells and Tissue.
The release from cells of signaling molecules through the controlled process of exocytosis involves multiple coordinated steps and is essential for the proper control of a multitude of biological pathways across the endocrine and nervous systems. However, these events are minute both temporally and in terms of the minute amounts of neurotransmitters, hormones, growth factors, and peptides released from single vesicles during exocytosis. It is therefore difficult to measure the kinetics of single exocytosis events in real time. One noninvasive method of measuring the release of molecules from cells is carbon-fiber amperometry. In this chapter, we will describe how we undertake such measurements from both single cells and in live tissue, how the subsequent data are analyzed, and how we interpret these results in terms of their relevant physiology.


Non-interventional LUME-BioNIS study of nintedanib plus docetaxel after chemotherapy in adenocarcinoma non-small cell lung cancer: A subgroup analysis in patients with prior immunotherapy.
To evaluate the effectiveness and safety of nintedanib plus docetaxel in patients with advanced adenocarcinoma non-small cell lung cancer (NSCLC) previously treated with both chemo- and immunotherapy. LUME-BioNIS is a European, prospective, multicenter, non-interventional study of patients with advanced adenocarcinoma NSCLC, who initiated nintedanib plus docetaxel after first-line chemotherapy in routine practice according to the approved nintedanib EU label. The primary objective is to explore whether molecular biomarkers can predict overall survival (OS). Information on clinical or radiologic progression and death, and adverse drug reactions (ADRs)/fatal adverse events (AEs) was collected during follow-up. Here, we report a subgroup analysis evaluating outcomes in immunotherapy-pretreated patients. Of 260 enrolled patients, 67 (25.8%) had prior immunotherapy and were included in this subgroup analysis. Prior immunotherapy was administered in first-line in 20 patients (29.9%; combined with chemotherapy in 4 patients [6.0%]) and later-lines in 47 patients (70.1%), and most commonly comprised nivolumab (39 patients; 58.2%), atezolizumab (14 patients; 20.9%) and pembrolizumab (11 patients; 16.4%). Nintedanib plus docetaxel was given in second-line in 10 patients (14.9%) and in later-lines in 57 patients (85.1%). Median OS was 8.8 months (95% confidence interval [CI]: 7.0-11.5) and median progression-free survival (PFS) was 4.6 months (95% CI: 3.5-5.7). Among 55 patients with available data, rates of objective response and disease control were 18.2% and 78.2%, respectively. In 65 patients evaluable for safety, the most common on-treatment ADRs/AEs were malignant neoplasm progression (19 patients; 29.2%), diarrhea (21 patients; 32.3%) and nausea (10 patients; 15.4%). Used according to the approved nintedanib label in routine practice, nintedanib plus docetaxel demonstrated clinical effectiveness, with no unexpected safety findings, in patients with prior chemotherapy and first- or later-line immunotherapy. These data add to the real-world evidence that can inform clinical decisions in the changing therapeutic landscape.


Fractionation of various elements in CRMs and in polluted soils.
An optimised BCR three steps sequential extraction procedure (BCR SEP) and several single extractions with KCl, NH(4)Cl, Na(4)P(2)O(7) and 0.5 mol L(-1) HCl were used for the fractionation of Al, Cu, Fe, Mn, Pb and Zn in CRMs and in samples from a mining area with sulphidic deposits. A good interlaboratory comparability was obtained for Cu, Pb and Zn in CRM 483, CRM 701, SRM 2710 and SRM 2711 by BCR SEP. The reliability of the results obtained is also very satisfactory. Some differences were found between our results and the indicative data for Al and Fe fractionation. However, serious discrepancies were found for Mn, not only for individual steps of the fractionation, but for the data obtained overall (sum of 1-3 steps), and for the total concentration as well. Our results could be utilized as a contribution to the existing indicative values for CRM 483, SRM 2710 and SRM 2711 for interlaboratory study. Moreover, data for the fractionation of elements mentioned above for CRM 701 are first presented here.A high correlation between 0.5 mol L(-1) of HCl-extractable amounts of the elements studied, and the sum of the three steps of BCR SEP in acid sulphatic weathering products and naturally acidified soils was established, which allows us to suggest this rapid and cost-effective single extraction procedure as a valuable tool in contamination assessment.


Microstructure and Defect-Based Fatigue Mechanism Evaluation of Brazed Coaxial Ti/Al2O3 Joints for Enhanced Endoprosthesis Design.
Alumina-based ceramic hip endoprosthesis heads have excellent tribological properties, such as low wear rates. However, stress peaks can occur at the point of contact with the prosthesis stem, increasing the probability of fracture. This risk should be minimized, especially for younger and active patients. Metal elevations at the stem taper after revision surgery without removal of a well-fixed stem are also known to increase the risk of fracture. A solution that also eliminates the need for an adapter sleeve could be a fixed titanium insert in the ceramic ball head, which would be suitable as a damping element to reduce the occurrence of stress peaks. A viable method for producing such a permanent titanium-ceramic joint is brazing. Therefore, a brazing method was developed for coaxial samples, and two modifications were made to the ceramic surface to braze a joint that could withstand high cyclic loading. This cyclic loading was applied in multiple amplitude tests in a self-developed test setup, followed by fractographic studies. Computed tomography and microstructural analyses-such as energy dispersive X-ray spectroscopy-were also used to characterize the process-structure-property relationships. It was found that the cyclic loading capacity can be significantly increased by modification of the surface structure of the ceramic.


Transcriptional inhibition of oxytocin receptor expression in human myometrial cells by melatonin involves protein kinase C signaling.
Our laboratory recently characterized the expression of the melatonin receptors in the human myometrium and showed that the expression of these receptors is suppressed during late pregnancy. In an effort to understand better the significance of melatonin in the human myometrium, we explored the mechanisms through which this hormone influences the expression of the oxytocin receptor in vitro. The stable melatonin analog iodomelatonin was presented to cultured telomerase-immortalized myometrial cells of the human telomerase reverse transcriptase line under physiological doses and durations. Pharmacological inhibitors of melatonin binding, gene transcription, phospholipase C, and protein kinase C signaling were used to define the mechanism of melatonin action. Our results reveal that melatonin significantly inhibits oxytocin receptor mRNA expression primarily via the melatonin 2 receptor. The melatonin-dependent decrease in oxytocin receptor transcripts involves suppression of gene transcription rather than enhanced rates of transcript degradation. Melatonin effects were abolished by pretreating the cells with the phospholipase C inhibitor U73122 or the protein kinase C inhibitor C1. Melatonin, like oxytocin, can negatively regulate oxytocin receptor transcription in human myometrial cells via modulation of protein kinase C signaling. This is consistent with the hypothesis that the reduced melatonin receptor expression during late pregnancy, which occurs at a time when oxytocin receptors are up-regulated, may be physiologically important for the subsequent timing of labor.


A study of the antidiarrheal properties of Loeselia mexicana on mice and rats.
Antidiarrheal properties of hexane, chloroform, methanol and aqueous extracts from Loeselia mexicana were studied using mice and rats as animal models. Flavones and sesquiterpenlactones were detected in the chemical screening of the aqueous extract. Diarrhea was induced by castor oil or MgSO4. The methanol extract diminished diarrhea in mice induced by MgSO4, while the aqueous extract showed an effect on castor-oil-induced. The aqueous extract also reduced castor-oil-enhanced intestinal transit and inhibited defecation of normal mice. The results obtained showed a symptomatic relief of diarrhea.


The learning curve of imageless robot-assisted total knee arthroplasty with standardised laxity testing requires the completion of nine cases, but does not reach time neutrality compared to conventional surgery.
The assistance of robot technology is introduced into the operating theatre to improve the precision of a total knee arthroplasty. However, as with all new technology, new technology requires a learning curve to reach adequate proficiency. The primary aim of this study was to identify the learning curve of an imageless robotic system with standardised laxity testing. The secondary aim of this study was to evaluate the accuracy of the intra-operative coronal alignment during the learning curve. A prospective study was performed on 30 patients undergoing robot-assisted total knee arthroplasty with an imageless robotic system (Corin, Massachusetts, USA) associated with a dedicated standardised laxity testing device. The learning curve of all surgical steps was assessed with intra-operative video monitoring. As comparison, the total surgical time of the last 30 patients receiving conventional total knee arthroplasty by the same surgeon and with the same implant was retrospectively assessed. Coronal lower limb alignment was evaluated pre- and post-operatively on standing full-leg radiographs. CUSUM (cumulative summation) analysis has shown inflexion points in multiple steps associated with robot-assisted surgery between one and 16 cases, which indicates the progression from the learning phase to the proficiency phase. The inflexion point for total operative time occurred after nine cases. Robot-assisted total knee surgery required significantly longer operative times than the conventional counterpart, with an average increase of 22 min. Post-operative limb and implant alignment was not influenced by a learning curve. The introduction of an imageless robotic system with standardised laxity assessment for total knee arthroplasty results in a learning curve of nine cases based on operative time. Compared to conventional surgery, the surgeon is not able to reach time neutrality with the robotic platform. There is no learning curve associated with coronal limb or implant alignment. This study enables orthopaedic surgeons to understand the implementation of this surgical system and its specific workflow into clinical practice.


Effects of two different meshes used in hernia repair on nerve transport.
We aimed to investigate the effects of two different types of mesh used in hernia repair on nerve transport and neural injury. Forty-five Wistar-albino rats were randomly allocated to three groups. Basal neural transport on the index of the left sciatic nerve was performed in all groups before surgery. In the control group, only sciatic nerve manipulation was performed. Other groups received a cuff around the index sciatic nerve. The second group received a monofilament polypropylene and the third group received a polytetrafluoroethylene mesh. Effects of entrapment on neural transport were evaluated by electromyography on the 28th day. Tissue samples from sciatic nerves were taken for evaluation of nerve injury. There was no significant difference between basal latency and amplitudes (p=0.609 and p=0.152, respectively). But latency was significantly different 4 weeks after the operation (p < 0.0001). At post hoc evaluation, entrapment groups had longer latency times compared with those in the control group (p=0.006 and p < 0.0001, respectively). But the increase in latency between entrapment arms was similar on the 28th day (p=0.601). Both initial and late term amplitudes were similar (p=0.364 and 0.913, respectively). Histologic evaluation by Masson's trichrome staining revealed high fibrosis scores and increased collagen deposits, especially in the polypropylene group (p < 0.0001), and increased inflammation in the polytetrafluoroethylene group (p < 0.001). Our results showed that the two most commonly used meshes in tension-free hernia surgery have different effects on nerve physiology and morphology. We concluded that alterations in nerve physiomorphology are from the properties of the mesh and may be the source of postoperative pain in hernia surgery.


New Technologies for Insect-Resistant and Herbicide-Tolerant Plants.
The advent of modern molecular biology and recombinant DNA technology has resulted in a dramatic increase in the number of insect-resistant (IR) and herbicide-tolerant (HT) plant varieties, with great economic benefits for farmers. Nevertheless, the high selection pressure generated by control strategies for weed and insect populations has led to the evolution of herbicide and pesticide resistance. In the short term, the development of new techniques or the improvement of existing ones will provide further instruments to counter the appearance of resistant weeds and insects and to reduce the use of agrochemicals. In this review, we examine some of the most promising new technologies for developing IR and HT plants, such as genome editing and antisense technologies.


Evaluation and comparison of nasal airway flow patterns among three subjects from Caucasian, Chinese and Indian ethnic groups using computational fluid dynamics simulation.
Nasal airflow is one of the most important determinants for nasal physiology. During the long evolution of human beings, different races have developed their own attributes of nasal morphologies which result in variations of nasal airflow patterns and nasal functions. This study evaluated and compared the effects of differences of nasal morphology among three healthy male subjects from Caucasian, Chinese and Indian ethnic groups on nasal airflow patterns using computational fluid dynamics simulation. By examining the anterior nasal airway, the nasal indices and the nostril shapes of the three subjects were found to be similar to nasal cavities of respective ethnic groups. Computed tomography images of these three subjects were obtained to reconstruct 3-dimensional models of nasal cavities. To retain the flow characteristics around the nasal vestibules, a 40 mm-radius semi sphere was assembled around the human face for the prescription of zero ambient gauge pressure. The results show that more airflow tends to pass through the middle passage of the nasal airway in the Caucasian model, and through the inferior portion in the Indian model. The Indian model was found with extremely low flow flux flowing through the olfactory region. The sizes of vortexes near the anterior cavity were found to be correlated with the angles between the upper nasal valve wall and the anterior head of the nasal cavity.


The structure of the vimentin gene.
The structure of the chromosomal gene encoding the intermediate filament protein vimentin is described. This gene, which is present as a single copy in the hamster genome, comprises about 10 kb of DNA and contains more than 80% of intron sequences. S1 mapping and sequence analysis reveal nine exons with a total length of 1848 nucleotides. For the complete primary structure of hamster vimentin, 464 amino acids are predicted, giving a molecular weight of 53,500 daltons. The intron positions are at codons 186, 206/207, 238/239, 292/293, 334/335, 408, 423, and 451/452. The overall homology with chicken desmin is 60% and is even higher in the central (alpha-helical) regions of both molecules. Cross-hybridization at the DNA level, however, is low. Comparison of the amino acid sequence of vimentin with prekeratin sequences shows that there is lesser homology of primary structure, but both the position and size of alpha-helical regions are strongly conserved. At the 5' end of the gene there is a consensus promoter sequence. The first AUG start codon is found 132 nucleotides downstream of the estimated cap site. The 3' nontranslated sequence shows homologies with the chicken vimentin gene. An interesting feature of the vimentin gene is a stretch of 44 nucleotides of alternating dC and dA within intron 2 that may form left-handed Z-DNA.


Update on Survival in Osteosarcoma.
Osteosarcoma is the most common primary bone malignancy in children. Treatment has evolved to include systemic chemotherapy and local control surgery. Although survival improved initially in a drastic fashion with this approach, recent decades have seen little to no further gains in this area. Limb salvage surgery evolved with effective chemotherapy and advances in imaging, and continues to improve in the recent era. This article serves as a review of survival in high-grade osteosarcoma: prognostic factors, advances in chemotherapy and surgery, late effects of chemotherapy and surgery in survivors, and future directions.


Tomotherapy PET-guided dose escalation: A dosimetric feasibility study for patients with malignant pleural mesothelioma.
The aim of this study was to investigate whether a safe escalation of the dose to the pleural cavity and PET/CT-positive areas in patients with unresectable malignant pleural mesothelioma (MPM) is possible using helical tomotherapy (HT). We selected 12 patients with MPM. Three planning strategies were investigated. In the first strategy (standard treatment), treated comprised a prescribed median dose to the planning target volume (PTV) boost (PTV1) of 64.5 Gy (range: 56 Gy/28 fractions to 66 Gy/30 fractions) and 51 Gy (range: 50.4 Gy/28 fractions to 54 Gy/30 fractions) to the pleura PTV (PTV2). Thereafter, for each patient, two dose escalation plans were generated prescribing 62.5 and 70 Gy (2.5 and 2.8 Gy/fraction, respectively) to the PTV1 and 56 Gy (2.24 Gy/fraction) to the PTV2, in 25 fractions. Dose-volume histogram (DVH) constraints and normal tissue complication probability (NTCP) calculations were used to evaluate the differences between the plans. For all plans, the 95 % PTVs received at least 95 % of the prescribed dose. For all patients, it was possible to perform the dose escalation in accordance with the Quantitative Analysis of Normal Tissue Effects in the Clinic (QUANTEC) constraints for organs at risk (OARs). The average contralateral lung dose was < 8 Gy. NTCP values for OARs did not increase significantly compared with the standard treatment (p > 0.05), except for the ipsilateral lung. For all plans, the lung volume ratio was strongly correlated with the V20, V30, and V40 DVHs of the lung (p < 0.0003) and with the lung mean dose (p < 0.0001). The results of this study suggest that by using HT it is possible to safely escalate the dose delivery to at least 62.5 Gy in PET-positive areas while treating the pleural cavity to 56 Gy in 25 fractions without significantly increasing the dose to the surrounding normal organs.


Effects of solvent polarity and solvent viscosity on the fluorescent properties of molecular rotors and related probes.
Fluorescent molecular rotors belong to a group of twisted intramolecular charge transfer complexes (TICT) whose photophysical characteristics depend on their environment. In this study, the influence of solvent polarity and viscosity on several representative TICT compounds (three Coumarin derivatives, 4,4-dimethylaminobenzonitrile DMABN, 9-(dicyanovinyl)-julolidine DCVJ), was examined. While solvent polarity caused a bathochromic shift of peak emission in all compounds, this shift was lowest in the case of molecular rotors. Peak intensity was influenced strongly by solvent viscosity in DMABN and the molecular rotors, but polarity and viscosity influences cannot be separated with DMABN. Coumarins, on the other hand, did not show viscosity sensitivity. This study shows the unique suitability of molecular rotors as fluorescent viscosity sensors.


Use of functional analysis methodology in the evaluation of medication effects.
The atypical antipsychotic medication risperidone was evaluated using a double-blind, placebo-controlled design in the treatment of destructive behavior in two individuals with autism. Pre-medication functional analyses indicated that destructive behavior was maintained by escape from demands, attention, or access to tangible items. For both individuals, destructive behavior during the demand condition was significantly reduced during the medication phases, whereas destructive behavior continued to occur to obtain tangible items (Reggie) and attention (Sean). In addition, there appeared to be a differential effect of the medication on self-injurious behavior (SIB) versus aggression for Sean. Results of the study demonstrate how functional analysis may provide information on those conditions and behaviors that are most likely to be affected by a specific medication.


[Application prospect of human-artificial intelligence system in future manned space flight].
To make the manned space flight more efficient and safer, a concept of human-artificial (AI) system is proposed in the present paper. The task of future manned space flight and the technique requirement with respect to the human-AI system development were analyzed. The main points are as follows: 1)Astronaut and AI are complementary to each other functionally; 2) Both symbol AI and connectionist AI should be included in the human-AI system, but expert system and Soar-like system are used mainly inside the cabin, the COG-like robots are mainly assigned for EVA either in LEO flight or on the surface of Moon or Mars; 3) The human-AI system is hierarchical in nature with astronaut at the top level; 4) The complex interfaces between astronaut and AI are the key points for running the system reliably and efficiently. As the importance of human-AI system in future manned space flight and the complexity of related technology, it is suggested that the R/D should be planned as early as possible.


Recombination between chloroplast genomes of Trachystoma ballii and Brassica juncea following protoplast fusion.
We document here the presence of a recombinant plastome in a cytoplasmic male sterile (CMS) line of Brassica juncea developed from the somatic hybrid Trachystoma ballii + B. juncea. Restriction endonuclease digestion of the chloroplast (cp) DNA has revealed that the recombinant plastome gives rise to novel fragments in addition to the parent-specific fragments. Analysis of the 16S rRNA region by Southern hybridization shows no variation between B. juncea, T. ballii and the CMS line. The rbcL gene region of the recombinant plastome is identical to that in T. ballii. Analysis with probes for psbA and psbD using single and double DNA digests indicates that the hybridization patterns of the recombinant plastome are identical to those of the parents in digests obtained with some restriction enzymes, while novel bands hybridize to probes in other digests. In the psbA region, a B. juncea-specific PstI site and a T. ballii-specific EcoRI site are found in the recombinant plastome. The pshD region of the recombinant plastome contains a B. juncea-specific HindIII site and T. ballii-specific BamHI and HpaII sites. These results indicate the occurrence of intergenomic recombination between the chloroplasts of T. ballii and B. juncea in the somatic hybrid from which the CMS line was developed. The recombined plastome appears to be a mosaic of fragments specific to both parents and the recombination event has occurred in the single-copy regions. These recombinational events have not caused any imbalance in the recombinant plastome in terms of chloroplast-related functions, which have remained stable over generations.


A comparison between liquid group A plasma and thawed group A plasma for massive transfusion activation in trauma patients.
The use of group A thawed 24-h plasma when resuscitating haemorrhagic shock patients has become more common; however, limited data exist on the clinical use of liquid plasma (LP). Our aim is to determine whether LP is of clinical benefit to patients requiring massive transfusion. The objective of this retrospective study was to detect any difference in 24-h survival between patients receiving liquid or thawed plasma (TP) during their massive transfusion activation. Other objectives were to report any difference in hospital length of stay (LOS), intensive care unit (ICU) LOS and in-hospital survival. Data collected included gender, age, mechanism of injury, Injury Severity Score, Revised Trauma Score and Trauma Injury Severity Score. A total of 178 patients received 1283 units of LP, median 4 and range (1-56), whereas 270 patients received 2031 units of TP, median 5 and range (1-87). The two study groups were comparable in terms of gender, age, mechanism of injury, whole blood, red blood cells, platelets and cryoprecipitate transfused. The use of LP during the massive transfusion activation in traumatically injured patients was not associated with increased 24-h survival compared to when using TP, p = 0.553. Our study did not show a difference in 24-h or 30-day survival between the use of LP compared to TP in trauma patients. LP should be considered an alternative to TP in trauma patients requiring immediate plasma resuscitation.


Trophoblastic cell sensitivity to 8-day chemotherapy in nonmetastatic gestational trophoblastic neoplasia.
Serial radioimmunoassay determinations of serum beta hCG and methotrexate were compared in two patients with nonmetastatic gestational trophoblastic neoplasia (NMGTN) treated with Goldstein's modification of Bagshawe's intermediate-dose methotrexate-citrovorum factor rescue-treatment program. Pretreatment beta hCG levels (mIU/ml) ranged within the outer limits of the 10(3) log level. Following intravenous methotrexate, sharp serum peaks between 10(-6) and 10(-5) M were observed. Plasma disappearance was rapid with a 3 log drop noted within 24 hr to levels incapable of inhibiting DNA synthesis. beta hCG levels manifested a 1 to 1.5 log drop over the 8 days of chemotherapy and complete remission was noted within 5 to 6 weeks of the first dose of methotrexate. No significant clinical or laboratory toxicity was observed. Although cell culture studies show that 100% of cell death can be achieved with serum levels of 10(-5) M in methotrexate-resistant choriocarcinoma, similar data do not exist for previously untreated trophoblastic neoplastic cells. These preliminary observations suggest that serum methotrexate levels are important for establishing sensitivity levels in a heterogeneous population of trophoblastic cells in NMGTN and that the total dose of methotrexate may be safely preselected on the basis of the pretreatment beta hCG.


"Candidatus Midichloriaceae" fam. nov. (Rickettsiales), an ecologically widespread clade of intracellular alphaproteobacteria.
"Candidatus Midichloria mitochondrii" is an intramitochondrial bacterium of the order Rickettsiales associated with the sheep tick Ixodes ricinus. Bacteria phylogenetically related to "Ca. Midichloria mitochondrii" (midichloria and like organisms [MALOs]) have been shown to be associated with a wide range of hosts, from amoebae to a variety of animals, including humans. Despite numerous studies focused on specific members of the MALO group, no comprehensive phylogenetic and statistical analyses have so far been performed on the group as a whole. Here, we present a multidisciplinary investigation based on 16S rRNA gene sequences using both phylogenetic and statistical methods, thereby analyzing MALOs in the overall framework of the Rickettsiales. This study revealed that (i) MALOs form a monophyletic group; (ii) the MALO group is structured into distinct subgroups, verifying current genera as significant evolutionary units and identifying several subclades that could represent novel genera; (iii) the MALO group ranks at the level of described Rickettsiales families, leading to the proposal of the novel family "Candidatus Midichloriaceae." In addition, based on the phylogenetic trees generated, we present an evolutionary scenario to interpret the distribution and life history transitions of these microorganisms associated with highly divergent eukaryotic hosts: we suggest that aquatic/environmental protista have acted as evolutionary reservoirs for members of this novel family, from which one or more lineages with the capacity of infecting metazoa have evolved.


RetroDISCO: Clearing technique to improve quantification of retrograde labeled motor neurons of intact mouse spinal cords.
Quantification of the number of axons reinnervating a target organ is often used to assess regeneration after peripheral nerve repair, but because of axonal branching, this method can overestimate the number of motor neurons regenerating across an injury. Current methods to count the number of regenerated motor neurons include retrograde labeling followed by cryosectioning and counting labeled motor neuron cell bodies, however, the process of sectioning introduces error from potential double counting of cells in adjacent sections. We describe a method, retroDISCO, that optically clears whole mouse spinal cord without loss of fluorescent signal to allow imaging of retrograde labeled motor neurons using confocal microscopy. Complete optical clearing of spinal cords takes four hours and confocal microscopy can obtain z-stacks of labeled motor neuron pools within 3-5min. The technique is able to detect anticipated differences in motor neuron number after cross-suture and conduit repair compared to intact mice and is highly repeatable. RetroDISCO is inexpensive, simple, robust and uses commonly available microscopy techniques to determine the number of motor neurons extending axons across an injury site, avoiding the need for labor-intensive cryosectioning and potential double counting of motor neuron cell bodies in adjacent sections. RetroDISCO allows rapid quantification of the degree of reinnervation without the confounding produced by axonal sprouting.


Capacity building in residential care.
A retrospective analysis of how the Dementia Care Program (DCP) contributed to capacity building at The Lodge at Broadmead (2004-2009) was conducted in 2010. Results showed facility-wide enhancements in (1) information capital, because of educational workshops and other forms of interactive learning for all staff, and (2) social capital, because of evaluation and information-sharing. It was concluded that the DCP increased staff's capacity to deliver best practice dementia care.


The development of a novel technique for predicting the exudate handling properties of modern wound dressings.
Numerous methods have been developed for measuring the absorbency of wound dressings but many are product specific and do not permit comparisons between different dressing types. Existing methods are also of limited value when assessing how a dressing is likely to perform in clinical practice. A new wound model has been developed that overcomes both of these problems. It facilitates direct comparisons between products that are different in structure and composition and provides, perhaps for the first time, a method that may be used to predict the time for which dressings might be expected to remain effective on exuding wounds. The historical background to the introduction of test methods for surgical dressings is briefly reviewed, and the development of the new wound model is described. Some preliminary results that illustrate the potential value of this new technique are also presented.


Environmental health risk assessment of dioxin in foods at the two most severe dioxin hot spots in Vietnam.
Bien Hoa and Da Nang airbases were bulk storages for Agent Orange during the Vietnam War and currently are the two most severe dioxin hot spots. This study assesses the health risk of exposure to dioxin through foods for local residents living in seven wards surrounding these airbases. This study follows the Australian Environmental Health Risk Assessment Framework to assess the health risk of exposure to dioxin in foods. Forty-six pooled samples of commonly consumed local foods were collected and analyzed for dioxin/furans. A food frequency and Knowledge-Attitude-Practice survey was also undertaken at 1000 local households, various stakeholders were involved and related publications were reviewed. Total dioxin/furan concentrations in samples of local "high-risk" foods (e.g. free range chicken meat and eggs, ducks, freshwater fish, snail and beef) ranged from 3.8 pg TEQ/g to 95 pg TEQ/g, while in "low-risk" foods (e.g. caged chicken meat and eggs, seafoods, pork, leafy vegetables, fruits, and rice) concentrations ranged from 0.03 pg TEQ/g to 6.1 pg TEQ/g. Estimated daily intake of dioxin if people who did not consume local high risk foods ranged from 3.2 pg TEQ/kg bw/day to 6.2 pg TEQ/kg bw/day (Bien Hoa) and from 1.2 pg TEQ/kg bw/day to 4.3 pg TEQ/kg bw/day (Da Nang). Consumption of local high risk foods resulted in extremely high dioxin daily intakes (60.4-102.8 pg TEQ/kg bw/day in Bien Hoa; 27.0-148.0 pg TEQ/kg bw/day in Da Nang). Consumption of local "high-risk" foods increases dioxin daily intakes far above the WHO recommended TDI (1-4 pg TEQ/kg bw/day). Practicing appropriate preventive measures is necessary to significantly reduce exposure and health risk.


A systematic diagnostic and therapeutic approach for the treatment of patients after cardio-pulmonary resuscitation: a prospective evaluation of 212 patients over 5 years.
A literature on systematic treatment protocols for patients after resuscitation for cardiac arrest is lacking. We evaluated a systematic protocol, including ECG, echocardiogram, urgent cardiac catheterisation ("STEMI-like" workflow), CT scans, laboratory findings, IABP, hypothermia, and cMRI, prospectively over 5 years. The primary endpoint was the Cerebral Performance Category Scale (CPCS). During the period from January 2008 to December 2012, 212 patients were included. The mean age was 66.7 years, n = 151 (71.2 %) were male, mean time from the first medical contact to start of catheterisation was 76.6 min, and ventricular fibrillation (VF) was present in n = 99 (46.7 %). A significant coronary artery stenosis was seen in n = 130 (61.3 %), PCI was performed in n = 101 (47.6 %), an ACS was found in n = 100 (47.2 %), n = 91 patients (42.9 %) had another cardiac cause, an extra-cardiac cause was found in n = 12 (5.7 %, mostly a cerebral process), and in 9 patients (4.3 %), no cause was identifiable. A significant difference in mortality was found for patients with TIMI flow 2/3 vs. 0/1 (65.4 vs. 95.7 %, p < 0.01). The difference of intra-aortic balloon pumping vs. no pumping was not significant, performing hypothermia reduced mortality significantly (52.7 vs. 68.2 %, p = 0.04). The survival rate was n = 76 (35.9 %), a CPCS of 1/2 was reached in n = 68 pts (32.1 %), patients with ongoing resuscitation had a 100 % mortality (n = 41), and VF had a lower mortality (54.6 vs. 72.6 %, p < 0.01). A systematic algorithm may improve the outcome of patients after reanimation compared with classically reported outcomes. The data are hypothesis generating for further studies.