Huntingtin Polyglutamine Fragments Are a Substrate for Hsp104 in Saccharomyces cerevisiae.
The aggregation of huntingtin fragments with expanded polyglutamine repeat regions (HttpolyQ) that cause Huntington's disease depends on the presence of a prion with an amyloid conformation in yeast. As a result of this relationship, HttpolyQ aggregation indirectly depends on Hsp104 due to its essential role in prion propagation. We find that HttQ103 aggregation is directly affected by Hsp104 with and without the presence of [RNQ+] and [PSI+] prions. When we inactivate Hsp104 in the presence of prion, yeast cells have only one or a few large HttQ103 aggregates rather than numerous smaller aggregates. When we inactivate Hsp104 in the absence of prion, there is no significant aggregation of HttQ103, whereas with active Hsp104, HttQ103 aggregates accumulate slowly due to the severing of spontaneously nucleated aggregates by Hsp104. We do not observe either effect with HttQ103P, which has a polyproline-rich region downstream of the polyglutamine region, because HttQ103P does not spontaneously nucleate and Hsp104 does not efficiently sever the prion-nucleated HttQ103P aggregates. Therefore, the only role of Hsp104 in HttQ103P aggregation is to propagate yeast prion. In conclusion, because Hsp104 efficiently severs the HttQ103 aggregates but not HttQ103P aggregates, it has a marked effect on the aggregation of HttQ103 but not HttQ103P.


Channel opening and gating mechanism in AMPA-subtype glutamate receptors.
AMPA (α-amino-3-hydroxy-5-methyl-4-isoxazole propionic acid)-subtype ionotropic glutamate receptors mediate fast excitatory neurotransmission throughout the central nervous system. Gated by the neurotransmitter glutamate, AMPA receptors are critical for synaptic strength, and dysregulation of AMPA receptor-mediated signalling is linked to numerous neurological diseases. Here we use cryo-electron microscopy to solve the structures of AMPA receptor-auxiliary subunit complexes in the apo, antagonist- and agonist-bound states and determine the iris-like mechanism of ion channel opening. The ion channel selectivity filter is formed by the extended portions of the re-entrant M2 loops, while the helical portions of M2 contribute to extensive hydrophobic interfaces between AMPA receptor subunits in the ion channel. We show how the permeation pathway changes upon channel opening and identify conformational changes throughout the entire AMPA receptor that accompany activation and desensitization. Our findings provide a framework for understanding gating across the family of ionotropic glutamate receptors and the role of AMPA receptors in excitatory neurotransmission.


Tautomerism and Rotamerism of Favipiravir and Halogenated Analogues in Solution and in the Solid State.
Favipiravir is an important selective antiviral against RNA-based viruses, and currently, it is being repurposed as a potential drug for the treatment of COVID-19. This type of chemical system presents different carboxamide-rotameric and hydroxyl-tautomeric states, which could be essential for interpreting its selective antiviral activity. Herein, the tautomeric 3-hydroxypyrazine/3-pyrazinone pair of favipiravir and its 6-substituted analogues, 6-Cl, 6-Br, 6-I, and 6-H, were fully investigated in solution and in the solid state through ultraviolet-visible, 1H nuclear magnetic resonance, infrared spectroscopy, and X-ray diffraction techniques. Also, a study of the gas phase was performed using density functional theory calculations. In general, the keto-enol balance in these 3-hydroxy-2-pyrazinecarboxamides is finely modulated by external and internal electrical variations via changes in solvent polarity or by replacement of substituents at position 6. The enol tautomer was prevalent in an apolar environment, whereas an increase in the level of the keto tautomer was favored by an increase in solvent polarity and, even moreso, with a strong hydrogen-donor solvent. Keto tautomerization was favored either in solution or in the solid state with a decrease in 6-substituent electronegativity as follows: H ≫ I ≈ Br > Cl ≥ F. Specific rotameric states based on carboxamide, "cisoide" and "transoide", were identified for the enol and keto tautomer, respectively; their rotamerism is dependent on the tautomerism and not the aggregation state.


Metal ions-binding T4 lysozyme as an intramolecular protein purification tag compatible with X-ray crystallography.
Phage T4 lysozyme is a well folded and highly soluble protein that is widely used as an insertion tag to improve solubility and crystallization properties of poorly behaved recombinant proteins. It has been used in the fusion protein strategy to facilitate crystallization of various proteins including multiple G protein-coupled receptors, lipid kinases, or sterol binding proteins. Here, we present a structural and biochemical characterization of its novel, metal ions-binding mutant (mbT4L). We demonstrate that mbT4L can be used as a purification tag in the immobilized-metal affinity chromatography and that, in many respects, it is superior to the conventional hexahistidine tag. In addition, structural characterization of mbT4L suggests that mbT4L can be used as a purification tag compatible with X-ray crystallography.


Role of hypoxia‑inducible factor‑1α in autophagic cell death in microglial cells induced by hypoxia.
Microglial cells are phagocytic cells of the central nervous system (CNS) and have been proposed to be a primary component of the innate immune response and maintain efficient CNS homeostasis. Microglial cells are activated during various phases of tissue repair and participate in various pathological conditions in the CNS. Following spinal cord injury (SCI), anoxemia is a key problem that results in tissue destruction. Hypoxia‑inducible factor 1‑α (HIF‑1α) may protect hypoxic cells from apoptosis or necrosis under ischemic and anoxic conditions. However, numerous studies have revealed that hypoxia upregulates HIF‑1α expression leading to the death of microglial cells. The present study investigated the alterations in HIF‑1α expression levels and the mechanism of autophagic cell death mediated by HIF‑1α in microglial cells induced by hypoxia. Hypoxia was demonstrated to induce HIF‑1α expression and autophagic cell death in microglial cells. Enhanced autophagy reduced cell death during the initial stages by restraining the functions of autophagy‑associated genes (microtubule‑associated protein 1A/1B‑light chain 3 phosphatidylethanolamine conjugate and Beclin‑1) and modulating the expression of inflammatory cytokines (tumor necrosis factor‑α and interleukin‑1β). Target value was determined by Cell Counting Kit 8 and cell death by flow cytometry. Transmission electron microscopy, immunohistochemical staining, reverse transcription‑quantitative polymerase chain reaction, western blotting, and ELISA were used for further analysis. However, increased expression of HIF‑1α induced cell death and autophagic cell death in microglial cells. Furthermore, the effects of the HIF‑1α inhibitor 2‑methoxyestradiol and HIF‑1α small interfering RNA on the death and autophagy of microglial cells in vitro were investigated. These investigations revealed the suppression of autophagy, the decrease of cell viability and the increase of inflammatory cytokines results from HIF‑1α inhibition or HIF‑1α silencing. In conclusion, the results indicated that appropriate expression of HIF‑1α can ameliorate autophagic cell death of microglial cells associated with hypoxia, and may provide a novel therapeutic approach for SCI associated with microglial cell activation.


Prenatal mood disturbance predicts sleep problems in infancy and toddlerhood.
Experimental animal data link prenatal stress with sleep disturbance in offspring, but the link in humans is unclear. To investigate the link between prenatal maternal anxiety and depression and infant sleep disturbance from 6 to 30 months of age. Longitudinal prospective study of a large birth cohort from pregnancy to 30 months. Questionnaire measures of anxiety and depression were completed by mothers at 18 and 32 weeks gestation and at 8 weeks and 8 months postpartum. The ALSPAC cohort, a prospective community study of women in the UK who have been followed since pregnancy. Measures of total sleep time, number of awakenings, and broadly defined sleep problems were available on children at ages 6, 18, and 30 months. Reliable measures of total sleep time, nighttime awakenings, and sleep problems were identified at 6, 18, and 30 months. Higher levels of prenatal maternal anxiety and depression predicted more sleep problems at 18 and 30 months, after controlling for postnatal mood and obstetric and psychosocial covariates; the association was not restricted to clinical extremes. No link with total sleep time was observed. Mood disturbance in pregnancy has persisting effects on sleep problems in the child, a finding that is consistent with experimental animal research. The findings add to a growing literature showing that maternal prenatal stress, anxiety, and depression may have lasting effects on child development.


An assessment of anthropometric indices and its association with NCDs among the older adults of India: evidence from LASI Wave-1.
The purpose of this study is to assess the status of physical body indices such as body mass index (BMI), waist circumference (WC), and waist-to-hip ratio (WHR) among the older adults aged 45 and above in India. Further, to explore the association of anthropometric indices with various non-communicable morbidities. The study uses secondary data of the Longitudinal Ageing Survey's first wave in India (2017-18). The national representative sample for older adults 45 and above (65,662) considered for the analysis. The prevalence of the non-communicable diseases (NCDs) included in the study is based on the self-reporting of the participants. Diseases included are among the top ten causes of death, such as cancer, hypertension, stroke, chronic heart diseases, diabetes, chronic respiratory diseases, and multi-morbidity. Multi-morbidity is a case of having more than one of the morbidities mentioned above. BMI-obese indicates an individual having a BMI ≥30, and the critical threshold value for high-risk WC for men is ≥102 cm while for women is ≥88 cm. The critical limit for the high-risk WHR for men and women is ≥0.90 and ≥ 0.85, respectively. Descriptive statistics and multiple logistic regressions are used to assess the association BMI, WC, and WHR with non-communicable morbidities. Based on the multivariate-adjusted model, odds shows that an Indian older adult aged 45 and above is 2.3 times more likely (AOR: 2.33; 95% CI (2.2, 2.5)) by obesity, 61% more likely (AOR: 1.61; 95% CI (1.629, 1.631)) by high-risk WHR and 98% more likely (AOR: 1.98; 95% CI (1.9, 2.1)) by high-risk WC to develop CVDs than their normal counterparts. Similarly, significant positive associations of obesity, high-risk WC, and high-risk WHR were observed with other NCDs and multi-morbidity. Our study shows that obesity, high-risk WC, and high-risk WHR are significant risks for developing NCDs and multi-morbidity among the older adults in India. There is a need for a multi-sectoral approach to reduce the share of the elderly population in high-risk groups of BMIs, WHR, and WC.


Two-year survey of etiologic agents of diarrheal disease at San Lazaro Hospital, Manila, Republic of the Philippines.
The prevalence of bacterial pathogens and rotavirus in 2,908 patients with diarrhea who were admitted to San Lazaro Hospital in Manila in 1983 and 1984 was determined. One or more enteric pathogens were isolated or detected in samples from 1,698 (58.4%) patients. Isolation rates for the various enteropathogens were as follows: rotavirus, 30.6%; Shigella spp., 11.6%; Salmonella spp., 9.2%; enterotoxigenic Escherichia coli (1983 only), 7.8%; Vibrio cholerae biotype eltor, 3.8%; non-O1 V. cholerae, 2.8%; Vibrio parahaemolyticus, 1.7%; other Vibrio spp., 1.1%; Campylobacter jejuni, 3.0%; Aeromonas hydrophila, 1.3%; and Plesiomonas shigelloides 1.1%. Giardia lamblia and Entamoeba histolytica were detected in 0.6 and 0.1%, respectively, of stool samples examined. Determination of the etiologic role of isolates was complicated by one or more of the following factors: isolation of multiple enteric pathogens (302 cases); isolation of Salmonella spp., enterotoxigenic E. coli, and C. jejuni from a similar proportion of asymptomatic control patients and patients with diarrhea; and isolation of a high proportion of certain pathogens (especially Salmonella spp.) only from enrichment broth, suggesting infection with a small number of organisms. Isolation of V. cholerae eltor was seasonal, with the majority of cases occurring in the rainy months. In addition, the number of patients with diarrhea increased with the onset of the monsoon rains and peaked during the months of maximum rainfall. Rotavirus infection occurred in both children and adults throughout the year and was the most frequently identified cause of diarrhea in children under 5 years of age. Shigella spp. were the most common agents of diarrhea in adults.


Effect of Conditioned Medium from Human Umbilical Cord-Derived Mesenchymal Stromal Cells on Rejuvenation of Nucleus Pulposus Derived Stem/Progenitor Cells from Degenerated Intervertebral Disc.
Mesenchymal stromal cells (MSCs)-based treatment for degeneration of intervertebral disc (IVD) has been proposed recently. We here addressed whether MSC secreted factors can rejuvenate nucleus pulposus- derived stem/progenitor cells from degenerated disc (D-NPSCs) in vitro. We analyzed the expression of MSCs and NP cell specific surface markers, pluripotency related genes, multilineage potential and cell proliferative capacity of D-NPSCs upon incubation with the conditioned medium which was collected from the umbilical cord derived MSCs (UCMSCs). Our results indicated that the conditioned medium restore the stemness of D-NPSCs by up-regulating the expression level of CD29 and CD105, pluripotency related genes OCT4 and Nanog, and NP progenitor marker Tie2. The increased stemness was accompanied by promoted cell proliferative capacity and improved osteogenic and chondrogenic differentiation potential. Our findings suggested that the UCMSCs derived conditioned medium might be used to rejuvenate the degenerated NP stem/progenitor cells.


Probing Variations of Reduction Activity at the Plasma Membrane Using a Targeted Ratiometric FRET Probe.
Plasma membrane (PM) is the turntable of various reactions that regulate essential functionalities of cells. Among these reactions, the thiol disulfide exchange (TDE) reaction plays an important role in cellular processes. We herein designed a selective probe, called membrane reduction probe (MRP), that is able to report TDE activity at the PM. MRP is based on a green emitting BODIPY PM probe connected to rhodamine through a disulfide bond. MRP is fluorogenic as it is turned off in aqueous media due to aggregation-caused quenching, and once inserted in the PM, it displays a bright red signal due to an efficient fluorescence energy resonance transfer (FRET) between the BODIPY donor and the rhodamine acceptor. In the PM model, the MRP can undergo TDE reaction with external reductive agents as well as with thiolated lipids embedded in the bilayer. Upon TDE reaction, the FRET is turned off and a bright green signal appears allowing a ratiometric readout of this reaction. In cells, the MRP quickly labeled the PM and was able to probe variations of TDE activity using ratiometric imaging. With this tool in hand, we were able to monitor variations of TDE activity at the PM under stress conditions, and we showed that cancer cell lines presented a reduced TDE activity at the PM compared to noncancer cells.


The Effect of Preoperative Neutrophil-To-Lymphocyte Ratio and Platelet-To-Lymphocyte Ratio on Predicting Rupture Risk in Tubal Ectopic Pregnancies.
Consecutive measurements of β-hCG levels and sonographic evaluation of adnexae are critical for choosing the optimal management in ampullar tubal ectopic pregnancies (EP). To select suitable patients for conservative approach, there is a need for an affordable and reliable marker for determining rupture risk. Evaluation of systemic inflammatory markers in combination with serum β-hCG levels and ultrasound might help to decide the appropriate treatment option. The purpose of the present study was to evaluate the diagnostic value of neutrophil-to-lymphocyte ratio (NLR) and platelet-to-lymphocyte ratio (PLR) in determining the rupture risk in ampullar tubal EPs and to compare with intraoperative findings. A total of 142 patients who underwent surgery for tubal EP were included. Seventy-two patients were in the intraoperatively diagnosed tubal rupture group and 70 patients without rupture findings were included in the control group. Both groups were compared for inflammation markers, β-hCG levels, and sonographic findings. Both NLR and PLR levels were found to be significantly higher in the tubal rupture group (4.62 ± 3.13 vs. 2.67 ± 1.43, 162.94 ± 63.61 vs. 115.84 ± 41.15, p < 0.01, respectively). According to the receiver operating characteristic analysis performed for the diagnostic performance of tubal diameter measurement, β-hCG, NLR, and PLR levels were significantly associated with histopathologically confirmed tubal rupture (p < 0.01). Systemic inflammatory markers are feasible and affordable tools for predicting tubal rupture risk in ampullar EPs and might be useful for determining surgery decision especially in low resource settings.


Unusual chemoselective Rh(II)-catalysed transformations of α-diazocarbonyl piperidine cores.
The reactivity of various α-diazocarbonyl piperidine scaffolds, characterised by an increased molecular complexity, was tested with various Rh(II) catalysts. The structure of the starting reagent is of relevance to the synthetic results. An unexpected dimerisation took place, starting from the simple piperidine scaffold, to give the hexahydrotetrazine ring system. Products derived from a nitrogen ylide intermediate or aromatic substitution (1,3,4,5-tetrahydro-2,5-methanobenzo[c]azepine and 1,2,3,3a-tetrahydrocyclopenta[de]isoquinolin-4(5 H)-one rings, respectively) were obtained from tetrahydroisoquinoline derivatives. The chemoselectivity of the reaction could be controlled by the choice of starting reagent, Rh(II) catalyst and the reaction conditions. Finally, it was found that the azepino heterocycle could coordinate to the catalyst to give new Rh(II) complexes.


Nanoemulsion: Concepts, development and applications in drug delivery.
Nanoemulsions are biphasic dispersion of two immiscible liquids: either water in oil (W/O) or oil in water (O/W) droplets stabilized by an amphiphilic surfactant. These come across as ultrafine dispersions whose differential drug loading; viscoelastic as well as visual properties can cater to a wide range of functionalities including drug delivery. However there is still relatively narrow insight regarding development, manufacturing, fabrication and manipulation of nanoemulsions which primarily stems from the fact that conventional aspects of emulsion formation and stabilization only partially apply to nanoemulsions. This general deficiency sets up the premise for current review. We attempt to explore varying intricacies, excipients, manufacturing techniques and their underlying principles, production conditions, structural dynamics, prevalent destabilization mechanisms, and drug delivery applications of nanoemulsions to spike interest of those contemplating a foray in this field.


Recovery from x-ray induced damage in human cells grown in culture.
Cells from an established human cell-line, NHIK 3025, originally derived from an early stage of cancer of the cervix, were tested for recovery capacity when irradiated with X-rays under aerobic (equilibrated with air) and extremely hypoxic conditions (O2 content less than 4 ppm). The dose-response curve obtained under aerobic conditions had a D0-value of 130 rads and an extrapolation number of 3.8. The corresponding curve obtained with a first dose exceeding that of the shoulder region of the survival curve followed 3 hours later by graded second doses, had a D0-value of 130 rads and extrapolation number 2.2. The curve obtained by measuring the survival after two equal doses of 330 rads, each with variable time intervals between the doses, showed a time dependent radiosensitivity. The dose-response curve obtained by irradiating the cells under extremely hypoxic conditions was well fitted by an exponential line up to about 2500 rads followed by a downward bend. Recovery from sublethal damage was not observed in split-dose experiments where the total doses were less than 2500 rads. For a total dose of 4200 rads, however, a split-dose effect was observed (SDR = 1.6). This split-dose effect is probably not due to Elkind repair, but is rather a consequence of the technique used.


Amyloid goiter: cytomorphological features and differential diagnosis on fine needle aspiration cytology: a case report.
Amyloid goiter is a rare cause of thyroid enlargement which can be confused clinically as well as cytologically with both colloid goiter and neoplastic process of thyroid. A 30-year-old man, diagnosed with chronic kidney disease 5 months previously and currently on dialysis and awaiting renal transplant, was referred by clinicians for fine needle aspiration cytology (FNAC) for thyroid swelling. FNAC showed dense amorphous clumps of extracellular material which appeared magenta colored on Giemsa and eosinophilic on Papanicolaou stain. Congo red staining and polarization showed characteristic apple green birefringence, thus confirming the material as amyloid, and the diagnosis of amyloid goiter was made. Amyloid on FNAC smears can be easily mistaken for colloid, and correct interpretation can avoid a false diagnosis of colloid goiter. A search should be made to look for any features suggestive of medullary carcinoma of the thyroid as amyloid is more often associated with it.


Catheter ablation of supraventricular tachycardia in patients with dextrocardia and situs inversus.
Dextrocardia with situs inversus is a rare cardiac positional anomaly. Catheter ablation procedures performed in this set of patients have not been sufficiently reported. A total of 10 patients with dextrocardia and situs inversus who received catheter ablation for supraventricular tachycardia (SVT) were included from a cohort of over 20 000 cases of catheter ablation for SVT in three centers from 2005 to 2016. All patients underwent electrophysiologic study and catheter ablation of SVT. Ablation targets were selected based on different tachycardia mechanisms with the primary endpoint of noninduction of tachycardia. The average age was 32.4 ± 5.6 years. Congenitally corrected transposition of great arteries (TGA) with situs inversus and D-looping of the ventricles and aorta (congenitally corrected TGA {I,D,D}) was found in four patients, while the other six patients exhibited mirror-image dextrocardia {I,L,L}. The mechanisms of SVT were atrioventricular nodal reentrant tachycardia in four patients, atrioventricular reentrant tachycardia in three, typical atrial flutter in one, intra-atrial reentrant tachycardia in one, and focal atrial tachycardia in one. Immediate procedural success was achieved in 9 out of 10 patients with no procedural complications. During a follow-up period of 6.3 ± 3.5 years on average, all patients remained free from recurrent tachycardia. For patients with dextrocardia and situs inversus, catheter ablation of SVT is safe and feasible. Differences in catheter maneuver and fluroscopy projection, along with difficulties in distorted anatomy are major obstacles for successful ablation.


Environmental dynamics during the onset of the Middle Stone Age in eastern Africa.
Development of the African Middle Stone Age (MSA) before 300,000 years ago raises the question of how environmental change influenced the evolution of behaviors characteristic of early Homo sapiens We used temporally well-constrained sedimentological and paleoenvironmental data to investigate environmental dynamics before and after the appearance of the early MSA in the Olorgesailie basin, Kenya. In contrast to the Acheulean archeological record in the same basin, MSA sites are associated with a markedly different faunal community, more pronounced erosion-deposition cycles, tectonic activity, and enhanced wet-dry variability. Aspects of Acheulean technology in this region imply that, as early as 615,000 years ago, greater stone material selectivity and wider resource procurement coincided with an increased pace of land-lake fluctuation, potentially anticipating the adaptability of MSA hominins.


Synthesis of 2-methyl-3-indolylacetic derivatives as anti-inflammatory agents that inhibit preferentially cyclooxygenase 1 without gastric damage.
Novel substituted 2-methyl-3-indolylacetic derivatives were synthesized and evaluated for their activity in vitro and in vivo on COX-1 and COX-2. Active compounds were screened to determine their gastrointestinal tolerability in vivo in the rat. Results showed that 3 and 4 preferentially inhibited COX-1 in vitro and in vivo. MD simulations indicated an induced fit for COX-1 but not for COX-2, probably because of a lower plasticity of the latter.


Computational formulation of a multiepitope vaccine unveils an exceptional prophylactic candidate against Merkel cell polyomavirus.
Merkel cell carcinoma (MCC) is a rare neuroendocrine skin malignancy caused by human Merkel cell polyomavirus (MCV), leading to the most aggressive skin cancer in humans. MCV has been identified in approximately 43%-100% of MCC cases, contributing to the highly aggressive nature of primary cutaneous carcinoma and leading to a notable mortality rate. Currently, no existing vaccines or drug candidates have shown efficacy in addressing the ailment caused by this specific pathogen. Therefore, this study aimed to design a novel multiepitope vaccine candidate against the virus using integrated immunoinformatics and vaccinomics approaches. Initially, the highest antigenic, immunogenic, and non-allergenic epitopes of cytotoxic T lymphocytes, helper T lymphocytes, and linear B lymphocytes corresponding to the virus whole protein sequences were identified and retrieved for vaccine construction. Subsequently, the selected epitopes were linked with appropriate linkers and added an adjuvant in front of the construct to enhance the immunogenicity of the vaccine candidates. Additionally, molecular docking and dynamics simulations identified strong and stable binding interactions between vaccine candidates and human Toll-like receptor 4. Furthermore, computer-aided immune simulation found the real-life-like immune response of vaccine candidates upon administration to the human body. Finally, codon optimization was conducted on the vaccine candidates to facilitate the in silico cloning of the vaccine into the pET28+(a) cloning vector. In conclusion, the vaccine candidate developed in this study is anticipated to augment the immune response in humans and effectively combat the virus. Nevertheless, it is imperative to conduct in vitro and in vivo assays to evaluate the efficacy of these vaccine candidates thoroughly. These evaluations will provide critical insights into the vaccine's effectiveness and potential for further development.


Age-distribution dynamics and aging indexes.
I analyze the dynamics of the age distribution as some vital rates change. When the fertility or mortality rate declines, the age distribution often manifests a dynamic stochastic-dominance relationship. I also propose some alternative indexes for measuring population aging. These indexes are closely connected with the age-distribution dynamics and contain more refined information about the distribution of age among the old.


Similar Comparative Low and High Doses of Deltamethrin and Acetamiprid Differently Impair the Retrieval of the Proboscis Extension Reflex in the Forager Honey Bee (Apis mellifera).
In the present study, the effects of low (10 ng/bee) and high (100 ng/bee) doses of acetamiprid and deltamethrin insecticides on multi-trial learning and retrieval were evaluated in the honey bee Apis mellifera. After oral application, acetamiprid and deltamethrin at the concentrations used were not able to impair learning sessions. When the retention tests were performed 1 h, 6 h, and 24 h after learning, we found a significant difference between bees after learning sessions when drugs were applied 24 h before learning. Deltamethrin-treated bees were found to be more sensitive at 10 ng/bee and 100 ng/bee doses compared to acetamiprid-treated bees, only with amounts of 100 ng/bee and at 6 h and 24 h delays. When insecticides were applied during learning sessions, none of the tested insecticides was able to impair learning performance at 10 ng/bee or 100 ng/bee but retention performance was altered 24 h after learning sessions. Acetamiprid was the only one to impair retrieval at 10 ng/bee, whereas at 100 ng/bee an impairment of retrieval was found with both insecticides. The present results therefore suggest that acetamiprid and deltamethrin are able to impair retrieval performance in the honey bee Apis mellifera.


Impact testing of the residual limb: System response to changes in prosthetic stiffness.
Currently, it is unknown whether changing prosthetic limb stiffness affects the total limb stiffness and influences the shock absorption of an individual with transtibial amputation. The hypotheses tested within this study are that a decrease in longitudinal prosthetic stiffness will produce (1) a reduced total limb stiffness, and (2) reduced magnitude of peak impact forces and increased time delay to peak force. Fourteen subjects with a transtibial amputation participated in this study. Prosthetic stiffness was modified by means of a shock-absorbing pylon that provides reduced longitudinal stiffness through compression of a helical spring within the pylon. A sudden loading evaluation device was built to examine changes in limb loading mechanics during a sudden impact event. No significant change was found in the peak force magnitude or timing of the peak force between prosthetic limb stiffness conditions. Total limb stiffness estimates ranged from 14.9 to 17.9 kN/m but were not significantly different between conditions. Thus, the prosthetic-side total limb stiffness was unaffected by changes in prosthetic limb stiffness. The insensitivity of the total limb stiffness to prosthetic stiffness may be explained by the mechanical characteristics (i.e., stiffness and damping) of the anatomical tissue within the residual limb.


Cell Surface Polysaccharides Represent a Common Strategy for Adsorption among Phages Infecting Lactic Acid Bacteria: Lessons from Dairy Lactococci and Streptococci.
Food fermentations rely on the application of robust bacterial starter cultures, the majority of which are represented by members of the lactic acid bacteria including Lactococcus lactis and Streptococcus thermophilus. Bacteriophage (or phage) proliferation remains one of the most significant threats to the fermentation industry. Therefore, it is imperative to define the phage ecology of fermented foods and to elucidate the mechanisms by which they recognize and bind to their bacterial hosts. Through a combination of functional and comparative genomics and structural analysis of the phage-host interactome, it is now possible to link the genotypes of strains of certain bacterial species to the chemical composition/structure of the associated cell wall polysaccharides (CWPS). In this paper, I discuss how the identification of common host recognition and binding strategies facilitates the development of rational starter culture systems and the implications of these findings in the context of sustainable food production systems.


[Relationship between the Trypsin Activity and Conformational Change Caused by Ultra High Static Pressure].
Trypsin was treated by high pressure technology, and its spatial structure was changed, the relationship between structural changes and trypsin activity was investigated. The secondary structure change of trypsin after pressure treatment was observed by Fourier transform infrared spectroscopy(FTIR). Moreover its tertiary structure change was observed by fluorescence spectroscopy; and its activity was tested using Folin phenol method. The results showed that, compared with the untreated(0.1 MPa), trypsin activity change was significant(p<0. 05) under different pressure(100~600 MPa) treatment at 37 °C for 20 min. After treated with 300 MPa, its activity was 0. 386 times higher than the untreated. Secondary structure of trypsin was analysed using FTIR, and the peak area ratio of α-helix and β-tum in secondary structure was the maximum(2. 749); Endogenous fluorescence spectra intensity was the maximum (1 353) at excitation wavelength 295 nm, and was 4 262 at excitation wavelength 280 nm; exogenous fluorescent spectra intensity was 2 022 at excitation wavelength 228 nm, all these change was remarkable(p<0. 05) comparing with the untreated. Therefore, ultrahigh pressure processing influence on the spatial structure of trypsin and induce enzyme activity.change. Trypsin activity is relate to the peak area ratio of α-helix and β-turn and the exposure degree of Trp and other hydrophobic a mino acid residues and Tyr.


Case report: A case of Culler-Jones syndrome caused by a novel mutation of GLI2 gene and literature review.
Culler-Jones syndrome is a rare clinical phenomenon with diverse manifestations and is prone to misdiagnosis. We report one patient who presented with a 10-year history of anosmia and a 1-year history of epididymal pain. Kallmann syndrome was suspected initially. The results of his laboratory tests, imaging, and genetic testing, however, combined to provide a conclusive diagnosis of Culler-Jones syndrome. With the aid of high-throughput sequencing technology, the GLI2 gene c.527A>G (p.Tyr176Cys) heterozygous mutation in the child was identified. No published works have yet described this mutation site. We described Culler-Jones syndrome in a child at length. We recommend that Culler-Jones syndrome be taken into account when considering the spectrum of disorders associated with abnormal growth and development in children. Once diagnosed, individualized hormone replacement treatment is required for each patient.


Brain Photobiomodulation-Preliminary Results from Regional Cerebral Oximetry and Thermal Imaging.
A new piece of equipment for LED (light emitting diode) brain photobiomodulation is introduced. Preliminary results from regional cerebral oxygen saturation and from thermography are shown before, during and after stimulation. The procedure offers a new way to quantify the biological effects of a possible innovative therapeutic method. However further measurements are absolutely necessary.


Primary cardiac angiosarcoma evaluated with contrast two-dimensional and real-time three-dimensional echocardiography.
A 63-year-old lady presented with syncope and atypical chest pain. Conventional two-dimensional (2D) contrast echocardiogram showed a highly vascular mass compressing the right atrium without direct communication. A real-time three-dimensional echocardiogram (RT3DE) revealed its outer spatial relationship with the surrounding structures and its inner heterogeneity as well. A chest computed tomography revealed a solid cardiac mass (6 x 7 cm(2)) arising from the right atrium and multiple pulmonary nodules. Histopathology of the pericardial fluid confirmed angiosarcoma. To our knowledge, this is the first report of histology-confirmed primary cardiac angiosarcoma, which was completely evaluated by conventional 2D, contrast, and RT3DE.


Deoxynivalenol affects in vitro intestinal epithelial cell barrier integrity through inhibition of protein synthesis.
Deoxynivalenol (DON), one of the most common mycotoxin contaminants of raw and processed cereal food, adversely affects the gastrointestinal tract. Since DON acts as a protein synthesis inhibitor, the constantly renewing intestinal epithelium could be particularly sensitive to DON. We analyzed the toxicological effects of DON on intestinal epithelial protein synthesis and barrier integrity. Differentiated Caco-2 cells, as a widely used model of the human intestinal barrier, were exposed to realistic intestinal concentrations of DON (50, 500 and 5000 ng/ml) during 24h. DON caused a concentration-dependent decrease in total protein content associated with a reduction in the incorporation of [(3)H]-leucine, demonstrating its inhibitory effect on protein synthesis. DON simultaneously increased the paracellular permeability of the monolayer as reflected through a decreased transepithelial electrical resistance associated with an increased paracellular flux of the tracer [(3)H]-mannitol. A concentration-dependent reduction in the expression level of the tight junction constituent claudin-4 was demonstrated by Western blot, which was not due to diminished transcription, increased degradation, or NF-kappaB, ERK or JNK activation, and was also observed for a tight junction independent protein, i.e. intestinal alkaline phosphatase. These results demonstrate a dual toxicological effect of DON on differentiated Caco-2 cells consisting in an inhibition of protein synthesis as well as an increase in monolayer permeability, and moreover suggest a possible link between them through diminished synthesis of the tight junction constituent claudin-4.


Liposomes as carriers of hydrophilic small molecule drugs: strategies to enhance encapsulation and delivery.
Although hydrophilic small molecule drugs are widely used in the clinic, their rapid clearance, suboptimal biodistribution, low intracellular absorption and toxicity can limit their therapeutic efficacy. These drawbacks can potentially be overcome by loading the drug into delivery systems, particularly liposomes; however, low encapsulation efficiency usually results. Many strategies are available to improve both the drug encapsulation efficiency and delivery to the target site to reduce side effects. For encapsulation, passive and active strategies are available. Passive strategies encompass the proper selection of the composition of the formulation, zeta potential, particle size and preparation method. Moreover, many weak acids and bases, such as doxorubicin, can be actively loaded with high efficiency. It is highly desirable that once the drug is encapsulated, it should be released preferentially at the target site, resulting in an optimal therapeutic effect devoid of side effects. For this purpose, targeted and triggered delivery approaches are available. The rapidly increasing knowledge of the many overexpressed biochemical makers in pathological sites, reviewed herein, has enabled the development of liposomes decorated with ligands for cell-surface receptors and active delivery. Furthermore, many liposomal formulations have been designed to actively release their content in response to specific stimuli, such as a pH decrease, heat, external alternating magnetic field, ultrasound or light. More than half a century after the discovery of liposomes, some hydrophilic small molecule drugs loaded in liposomes with high encapsulation efficiency are available on the market. However, targeted liposomes or formulations able to deliver the drug after a stimulus are not yet a reality in the clinic and are still awaited.


An Improved UWB/IMU Tightly Coupled Positioning Algorithm Study.
The combination of ultra-wide band (UWB) and inertial measurement unit (IMU) positioning is subject to random errors and non-line-of-sight errors, and in this paper, an improved positioning strategy is proposed to address this problem. The Kalman filter (KF) is used to pre-process the original UWB measurements, suppressing the effect of range mutation values of UWB on combined positioning, and the extended Kalman filter (EKF) is used to fuse the UWB measurements with the IMU measurements, with the difference between the two measurements used as the measurement information. The non-line-of-sight (NLOS) measurement information is also used. The optimal estimate is obtained by adjusting the system measurement noise covariance matrix in real time, according to the judgment result, and suppressing the interference of non-line-of-sight factors. The optimal estimate of the current state is fed back to the UWB range value in the next state, and the range value is dynamically adjusted after one-dimensional filtering pre-processing. Compared with conventional tightly coupled positioning, the positioning accuracy of the method in this paper is improved by 46.15% in the field experimental positioning results.


Anisotropic multi-scale fluid registration: evaluation in magnetic resonance breast imaging.
Registration using models of compressible viscous fluids has not found the general application of some other techniques (e.g., free-form-deformation (FFD)) despite its ability to model large diffeomorphic deformations. We report on a multi-resolution fluid registration algorithm which improves on previous work by (a) directly solving the Navier-Stokes equation at the resolution of the images, (b) accommodating image sampling anisotropy using semi-coarsening and implicit smoothing in a full multi-grid (FMG) solver and (c) exploiting the inherent multi-resolution nature of FMG to implement a multi-scale approach. Evaluation is on five magnetic resonance (MR) breast images subject to six biomechanical deformation fields over 11 multi-resolution schemes. Quantitative assessment is by tissue overlaps and target registration errors and by registering using the known correspondences rather than image features to validate the fluid model. Context is given by comparison with a validated FFD algorithm and by application to images of volunteers subjected to large applied deformation. The results show that fluid registration of 3D breast MR images to sub-voxel accuracy is possible in minutes on a 1.6 GHz Linux-based Athlon processor with coarse solutions obtainable in a few tens of seconds. Accuracy and computation time are comparable to FFD techniques validated for this application.


Tubular epithelial cell dedifferentiation is driven by the helix-loop-helix transcriptional inhibitor Id1.
In the fibrotic kidney, tubular cells undergo epithelial-to-mesenchymal transition (EMT), a phenotypic conversion that is characterized by sequential loss of epithelial markers and gain of mesenchymal features. For understanding of the molecular mechanism that governs this process, a high-throughput gene expression microarray analysis was used to identify the critical genes in the initial phase of the TGF-beta1-mediated EMT. Inhibitor of differentiation-1 (Id1), a dominant negative antagonist of the basic helix-loop-helix transcription factors, was found to be induced rapidly in human proximal tubular epithelial cells after TGF-beta1 treatment. This induction of Id1 depended on intracellular Smad signaling. Ectopic expression of Id1 suppressed epithelial E-cadherin and zonula occludens-1 expression. Id1 physically formed complex with basic helix-loop-helix transcription factor HEB (Hela E-box binding factor), sequestered its ability to bind to E-box, and repressed the trans-activation of E-cadherin promoter. However, overexpression of Id1 failed to induce alpha-smooth muscle actin, matrix metalloproteinase-2, fibronectin, and integrin-linked kinase (ILK), indicating its inability to confer a complete EMT. Overexpression of ILK or inhibition of ILK activity had no effect on Id1 induction by TGF-beta1, suggesting that Id1 and ILK have independent roles in epithelial dedifferentiation and EMT. In vivo, Id1 was induced exclusively in the degenerated, dilated renal tubular epithelium after unilateral ureteral obstruction. These studies identify Id1 transcriptional inhibitor as a crucial player in mediating cell dedifferentiation of renal tubular epithelium and suggest that EMT is a multistep process in which loss of epithelial adhesion does not necessarily lead to an autonomous mesenchymal transition.


Spatiotemporal Heterogeneity of Lung-Deposited Surface Area in Zurich Switzerland: Lung-Deposited Surface Area as a New Routine Metric for Ambient Particle Monitoring.
Objective: To assess the spatiotemporal heterogeneity of lung-deposited particle surface area concentration (LDSA), while testing the long-term performance of a prototype of low-cost-low-maintenance LDSA sensors. One factor hampering epidemiological studies on fine to ultrafine particles (F-to-UFP) exposure is exposure error due to their high spatiotemporal heterogeneity, not reflected in particle mass. Though LDSA shows consistent associations between F-to-UFP exposure and health effects, LDSA data are limited. Methods: We measured LDSA in a network of ten sensors, including urban, suburban, and rural environments in Zurich, Switzerland. With traffic counts, traffic co-pollutant concentrations, and meteorological parameters, we assessed the drivers of the LDSA observations. Results: LDSA reflected the high spatiotemporal heterogeneity of F-to-UFP. With micrometeorological influences, local sources like road traffic, restaurants, air traffic, and residential combustion drove LDSA. The temporal pattern of LDSA reflected that of the local sources. Conclusion: LDSA may be a viable metric for inexpensively characterizing F-to-UFP exposure. The tested devices generated sound data and may significantly contribute to filling the LDSA exposure data gap, providing grounds for more statistically significant epidemiological studies and regulation of F-to-UFP.


Evaluation of European-based polygenic risk score for breast cancer in Ashkenazi Jewish women in Israel.
Polygenic risk score (PRS), calculated based on genome-wide association studies (GWASs), can improve breast cancer (BC) risk assessment. To date, most BC GWASs have been performed in individuals of European (EUR) ancestry, and the generalisation of EUR-based PRS to other populations is a major challenge. In this study, we examined the performance of EUR-based BC PRS models in Ashkenazi Jewish (AJ) women. We generated PRSs based on data on EUR women from the Breast Cancer Association Consortium (BCAC). We tested the performance of the PRSs in a cohort of 2161 AJ women from Israel (1437 cases and 724 controls) from BCAC (BCAC cohort from Israel (BCAC-IL)). In addition, we tested the performance of these EUR-based BC PRSs, as well as the established 313-SNP EUR BC PRS, in an independent cohort of 181 AJ women from Hadassah Medical Center (HMC) in Israel. In the BCAC-IL cohort, the highest OR per 1 SD was 1.56 (±0.09). The OR for AJ women at the top 10% of the PRS distribution compared with the middle quintile was 2.10 (±0.24). In the HMC cohort, the OR per 1 SD of the EUR-based PRS that performed best in the BCAC-IL cohort was 1.58±0.27. The OR per 1 SD of the commonly used 313-SNP BC PRS was 1.64 (±0.28). Extant EUR GWAS data can be used for generating PRSs that identify AJ women with markedly elevated risk of BC and therefore hold promise for improving BC risk assessment in AJ women.


Optimal standard measures: Comment on Matthews et al. (2016).
Matthews, Adler, Forrest, and Stead (2016) implied that measures of mental health constructs such as depression can be made interchangeable by standardizing their scores. The present author contends that content differences in the measures a priori prevent their equivalence and that, instead, we need to agree on a standard definition of the construct and then use only a definition-fitting optimal standard measure. (PsycINFO Database Record


Tolerance and efficacy of palliative radiotherapy for advanced pancreatic cancer: A retrospective analysis of single-institutional experiences.
This study was conducted to investigate hypofractionated radiotherapy (RT) in patients with locally advanced or metastatic adenocarcinoma of the pancreas. A total of 31 patients were enrolled in this study, 26 of whom had locally advanced (M0) pancreatic cancer and 5 had metastatic (M1) disease. The patients were treated with palliative RT (6-30 Gy in 1-10 fractions over a period of 1 day-2 weeks). Treatment-related toxicity was classified according to the Common Terminology Criteria for Adverse Events, version 3.0. Early mild toxicity was observed. A total of 17 patients (55%) achieved good pain control without pharmacological therapy, and 12 patients (39%) reduced their use of analgesics; in the remaining 2 patients (6%), there was no change in analgesic use. Late high-grade (>3) toxicity was not observed. The average survival time for the 31 patients was 9 months. The 1-year overall survival rate was 16%. Palliative RT was well-tolerated and was able to prolong the survival time. The majority of the patients achieved better pain control with palliative RT.


Pilot In-Flight Sleep During Long-Range and Ultra-Long Range Commercial Airline Flights.
INTRODUCTION: In commercial aviation, pilot fatigue is a major threat to safety. One key fatigue mitigation strategy on long-range (LR; 8-16 h) and ultra-long range (ULR; 16+ h on at least 10% of trips) routes is allotting in-flight rest breaks for the pilots. Since sleep is a strong predictor of performance, it is important to quantify total in-flight sleep (TIFS) and determine rest scheme schedules that optimize sleep opportunity and subsequent performance. Here we quantify in-flight sleep and characterize rest schemes by type and efficiency.METHODS: Between 2015 and 2019, we collected data on in-flight sleep on 3 LR and 5 ULR routes totaling 231 pilots flying over 1200 flight duty periods. Data were collected using a combination of actigraphy and logbooks.RESULTS: Over all combinations of flight direction, crew and LR vs. ULR, average TIFS ranged from 3.4 h to 5.2 h with some ULR pilots getting over 8 h. Most crews made use of simple two- or three-break rest schemes and the complex four-break rest schemes were used almost exclusively on the three longest ULR routes. The complex schemes were less efficient than simple schemes, although this effect was small. Complex schemes resulted in no more TIFS compared to simple schemes on the same routes.DISCUSSION: Overall, we find that crews are getting more sleep on these routes than previously reported on similar routes. Most crews use simple rest schemes and these simple schemes are more efficient than complex schemes.Rempe MJ, Basiarz E, Rasmussen I, Belenky G, Lamp A. Pilot in-flight sleep during long-range and ultra-long range commercial airline flights. Aerosp Med Hum Perform. 2022; 93(4):368-375.


Synthesis, characterization, and detection of new oxandrolone metabolites as long-term markers in sports drug testing.
The discovery and implementation of the long-term metabolite of metandienone, namely 17β-hydroxymethyl-17α-methyl-18-norandrost-1,4,13-trien-3-one, to doping control resulted in hundreds of positive metandienone findings worldwide and impressively demonstrated that prolonged detection periods significantly increase the effectiveness of sports drug testing. For oxandrolone and other 17-methyl steroids, analogs of this metabolite have already been described, but comprehensive characterization and pharmacokinetic data are still missing. In this report, the synthesis of the two epimeric oxandrolone metabolites-17β-hydroxymethyl-17α-methyl-18-nor-2-oxa-5α-androsta-13-en-3-one and 17α-hydroxymethyl-17β-methyl-18-nor-2-oxa-5α-androsta-13-en-3-one-using a fungus (Cunninghamella elegans) based protocol is presented. The reference material was fully characterized by liquid chromatography nuclear magnetic resonance spectroscopy and high resolution/high accuracy mass spectrometry. To ensure a specific and sensitive detection in athlete's urine, different analytical approaches were followed, such as liquid chromatography-tandem mass spectrometry (QqQ and Q-Orbitrap) and gas chromatography-tandem mass spectrometry, in order to detect and identify the new target analytes. The applied methods have demonstrated good specificity and no significant matrix interferences. Linearity (R(2) > 0.99) was tested, and precise results were obtained for the detection of the analytes (coefficient of variation <20%). Limits of detection (S/N) for confirmatory and screening analysis were estimated at 1 and 2 ng/mL of urine, respectively. The assay was applied to oxandrolone post-administration samples to obtain data on the excretion of the different oxandrolone metabolites. The studied specimens demonstrated significantly longer detection periods (up to 18 days) for the new oxandrolone metabolites compared to commonly targeted metabolites such as epioxandrolone or 18-nor-oxandrolone, presenting a promising approach to improve the fight against doping.


Stability of EPSPS gene copy number in Hordeum glaucum Steud (barley grass) in the presence and absence of glyphosate selection.
Gene amplification has been shown to provide resistance to glyphosate in several weed species, including Hordeum glaucum populations in South Australia. The stability of 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) gene copies in resistant populations in the presence or absence of glyphosate selection has not been determined. Applying glyphosate to a cloned plant resulted in an increase in resistance and EPSPS copy number in the progeny of that plant compared to the untreated clone. The LD50 (herbicide concentration required for 50% mortality) increased by 75% to 79% in the progeny of the treated clones compared to the untreated in both populations (YP-17 and YP-16). EPSPS copy number estimates were higher in treated individuals compared to untreated individuals with an average of seven copies compared to six in YP-16 and 11 compared to six in YP-17. There was a positive correlation (R2 = 0.78) between EPSPS copy number and LD50 of all populations. EPSPS gene copy number and resistance to glyphosate increased in H. glaucum populations under glyphosate selection, suggesting the number of EPSPS gene copies present is dependent on glyphosate selection. © 2021 Society of Chemical Industry.


Day Bed Unit at the Department of Gastro-enterology. Experience of a study group.
Day Bed Unit at the Department of Gastroenterology. Experience of a study group. We analyzed the activity of the Day-Bed Unit at the Department of Gastroenterology of Turin Hospital (Molinette). The quality of the service provided and the days of stay in hospital for each admission were evaluated in terms of cost benefit ratio. The average stay in hospital for each admission was 2.57 days as opposed to 9.3 in the in patient ward. Patients were divided in subsets according to the cause of admission. Stratifying the patients according to diagnosis showed a hospital stay of 2.73 days for liver disease and of 2.81 for pancreas and biliary disease. Among the admissions for liver disease, pre or posttranspiant patients required 2.89 days as opposed to 2.62 days of those who were treated for esophageal varices. Thus, the cause of admission was a factor influencing length of hospital stay. Patients needing non surgical treatment for liver cancer took 2.22 days if treated with percutaneous ethanol injection; those undergoing chemoembolization required 2.93 days. In conclusion, the day bed unit has proven to be able to provide a service with a good cost benefit ratio. Patients admitted to this service may be withdrawn from the waiting list of the in patient ward, thus reducing the waiting time. Optimization of this service needs integration with the surroudings Units within the Department and in the Hospital.


[Violation of the homologous chromosomes approchement as the answer to the small doses of roentgen irradiation].
Any violation of the homologous chromosome DNA repair leads to the genome instability, characteristic for hereditary syndromes and for aging cells. Using low doses of ionizing radiation (3-10 cGy) we have found any transference of the homologous centromere loci of the chromosome 1 (1q12) from the periphery to the cen- tre of the nucleus in the lymphocytes of young healthy donors. The same effect was found after any influence of RNA-polymerase inhibitor a-amanitine. Some changes in the chromatin structure during aging (70-80 years old patients) result in the difficulties in chromosome displacement, accompanied with any trouble in the appro- achement of the homologous chromosome loci as an answer to low doses of radiation.


Bottleneck effect in three-dimensional turbulence simulations.
At numerical resolutions around 512(3) and above, three-dimensional energy spectra from turbulence simulations begin to show noticeably shallower spectra than k(-5/3) near the dissipation wave number ("bottleneck effect"). This effect is shown to be significantly weaker in one-dimensional spectra such as those obtained in wind tunnel turbulence. The difference can be understood in terms of the transformation between the one-dimensional and three-dimensional energy spectra under the assumption that the turbulent velocity field is isotropic. Transversal and longitudinal energy spectra are similar and can both accurately be computed from the full three-dimensional spectra. Second-order structure functions are less susceptible to the bottleneck effect and may be better suited for inferring the scaling exponent from numerical simulation data.


Variability in flexible sigmoidoscopy performance among examiners in a screening trial.
The efficacy of flexible sigmoidoscopy (FSG) in reducing colorectal cancer mortality is being evaluated in randomized trials. In 2 European trials, wide variability across examiners in FSG performance was noted. We report on the performance of examiners in the US randomized trial: the Prostate, Lung, Colorectal and Ovarian Cancer Screening Trial. Screening was performed at 10 geographically dispersed clinical centers. Patients with screens positive for a lesion or mass were referred to their private health care providers for endoscopic follow-up evaluation; lesions were not removed and a biopsy examination was not performed at screening. FSG performance among 64 examiners at these centers, each performing 100 or more baseline FSG examinations, with an aggregate of almost 50,000 examinations, was analyzed. Screen-positivity results among examiners ranged from 9%-58%, with a coefficient of variation (CV) of 36%. CVs were 29% for distal polyp detection and 21% for distal adenoma detection. Inadequate rates ranged from 1%-27% (CV, 52%). Examiners with higher screen-positivity rates had higher false-positive rates, defined as a positive screen with no distal lesion found on endoscopic follow-up evaluation. Considerable variability exists in the rates of positive screens and in polyp and adenoma detection rates among FSG examiners performing the procedures using a common protocol.


New Frontiers in Vascular Access Practice: From Standardized to Patient-tailored Care and Shared Decision Making.
Vascular access planning is critical in the management of patients with advanced kidney disease who elect for hemodialysis for RRT. Policies put in place more than two decades ago attempted to standardize vascular access care around the model of optimal, namely arteriovenous fistula, and least preferred, namely central venous catheter, type of access. This homogenized approach to vascular access care emerged ineffective in the increasingly heterogeneous and complex dialysis population. The most recent vascular access guidelines acknowledge the limitations of standardized care and encourage tailoring vascular access care on the basis of patient and disease characteristics. In this article, we discuss available literature in support of patient-tailored access care on the basis of differences in vascular access outcomes by biologic and social factors-age, sex, and race. Further, we draw attention to the overlooked dimension of patient-reported preferences and shared decision making in the practice of vascular access planning. We discuss milestones to overcome as requisite steps to implement effective shared decision making in vascular access care. Finally, we take into consideration local practice cofactors as major players in vascular access fate. We conclude that a personalized approach to hemodialysis vascular access will require dynamic care specifically relevant to the individual on the basis of biologic factors, fluctuating clinical needs, values, and preferences.


Effects of heat treatment on metal-ceramic combination of selective-laser-melted cobalt-chromium alloy.
Components fabricated by selective laser melting (SLM) deform because of residual stress, but heat treatment allows the release of that stress and avoids deformation. Although dental cobalt-chromium (Co-Cr) alloy has been specifically designed for SLM, the effects of heat treatment on the metal-ceramic combination of SLM Co-Cr restorations require investigation. The purpose of this in vitro study was to evaluate the effects of heat treatment on the metal-ceramic combination of SLM Co-Cr alloy. Following ISO 9693:2012, Co-Cr metal strips (Solibond C Plus cast alloy and SLM powders; YETI Dental) were fabricated with a dimension of 25×3×0.5 mm by casting and SLM. The SLM specimens were divided into 3 subgroups (n=15 for each subgroup). Two subgroups were subjected to heat treatment at 880°C (SLM-880) and 1100°C (SLM-1100). The third subgroup was not subjected to heat treatment and served as a control (As-SLM). Cast specimens (n=15) also acted as a control. A porcelain layer with a thickness of 1.1 mm was fired to the central area (3×8 mm) of each specimen. The 3-point bend test was used to evaluate the metal-ceramic bond strength (τb). The fractured metal surfaces were examined by the naked eye, using a digital camera, and also using a scanning electron microscope. The area fraction of adherence porcelain (AFAP) was determined by measuring the atomic percentage of silicon using energy-dispersive x-ray spectroscopy (EDS). One-way ANOVA and the Kruskal-Wallis test, followed by the Mann-Whitney test (α=.05), were used for statistical analysis. The 1-way ANOVA found no significant difference in the bond strength among the 4 groups. The EDS analysis indicated that specimens in the groups receiving heat treatment (SLM-880 and SLM-1100) showed higher AFAP values than those in the As-SLM group (P<.05). The SLM-880 also showed significantly higher AFAP values than the SLM-1100. Compared with the cast group, significant differences in AFAP values were also observed, and the specimens in the order of highest to lowest AFAP values were SLM-880>cast>As-SLM; no significant difference was found between the SLM-1100 and cast groups. Heat treatment at 880°C and 1100°C did not affect the metal-ceramic bond strength of Co-Cr alloy made by SLM but did improve the porcelain adherence. SLM-fabricated and heat-treated Co-Cr alloy shows comparable or more porcelain adherence than cast specimens.


Arterial percutaneous angioplasty in upper limbs with vascular access devices for haemodialysis.
The purpose of this study was to evaluate retrospectively the clinical context and effectiveness of arterial percutaneous transluminal angioplasty (PTA) of arterio-venous fistulae in chronic haemodialysis patients. Between May 1992 and June 1997, arterial PTA was performed in 33 patients with a total of 35 angioaccess devices of the upper limbs (18 arterio-venous fistulae and 17 PTFE grafts). Clinical indications for arterial PTA were unexplained acute thrombosis in 12 patients (34.3%), insufficient blood flow in 13 patients (37.1%), and severe limb ischaemia in 10 patients (28.6%), two of whom had skin ulcerations and one had severe neurological damage. Follow-up periods varied between 1 and 55 months (mean 15.5 months). PTA was attempted in 22 radial, 10 brachial and seven ulnar arteries. Angioplasty was successful (i.e. residual stenosis of </=30%) in all but one patient. There were no complications. Early re-thrombosis (<1 month) occurred in two of the 12 patients with acute occlusions. All the angioaccesses of patients with insufficient blood flow were improved. Eight of the patients with limb ischaemia became symptom free, and two were failures (one had partial healing of skin ulcerations and one did not improve). Re-stenosis occurred in six cases (27.3% of the 22 angiograms performed) but re-dilatation was performed in only two instances. Primary and secondary patencies were 63.5 and 90.6% at 6 months and 40.8 and 75.6% at 24 months, respectively. Chronic arterial lesions in upper limbs bearing vascular access devices for haemodialysis may lead to thrombosis, ischaemia and insufficient flow for dialysis treatment. PTA is a safe and effective technique with a low rate of re-intervention.


Diversity in medullary thymic epithelial cells controls the activity and availability of iNKT cells.
The thymus supports multiple αβ T cell lineages that are functionally distinct, but mechanisms that control this multifaceted development are poorly understood. Here we examine medullary thymic epithelial cell (mTEC) heterogeneity and its influence on CD1d-restricted iNKT cells. We find three distinct mTEClow subsets distinguished by surface, intracellular and secreted molecules, and identify LTβR as a cell-autonomous controller of their development. Importantly, this mTEC heterogeneity enables the thymus to differentially control iNKT sublineages possessing distinct effector properties. mTEC expression of LTβR is essential for the development thymic tuft cells which regulate NKT2 via IL-25, while LTβR controls CD104+CCL21+ mTEClow that are capable of IL-15-transpresentation for regulating NKT1 and NKT17. Finally, mTECs regulate both iNKT-mediated activation of thymic dendritic cells, and iNKT availability in extrathymic sites. In conclusion, mTEC specialization controls intrathymic iNKT cell development and function, and determines iNKT pool size in peripheral tissues.


Data-Driven Residency Training: A Scoping Review of Educational Interventions for Neurosurgery Residency Programs.
Neurosurgery residency programs are tasked with imparting large volumes of both clinical knowledge and technical skill to trainees in limited time. Many investigators have described local practices, which may offer evidence-based interventions in neurosurgical residency education, but this literature has not been systematically reviewed. To perform a scoping review of educational practices in neurosurgery, which are supported by quantitative, peer-reviewed research. A scoping review of the literature was performed. PubMed, Embase, and Web of Science databases were queried for articles describing educational interventions for neurosurgery residents, which included a quantitative assessment of the effect on resident performance. From an initial set of 1785 unique articles, 29 studies were ultimately screened and included. Studies were into the following 6 topics: (1) didactics and curricula (n = 13), (2) nontechnical skills (n = 6), (3) wellness and burnout (n = 4), (4) assessment and feedback (n = 2), (5) mentorship and career development (n = 2), and (6) research (n = 2). Individual study results were described. Several educational interventions in neurosurgical training are supported by quantitative evidence. Methodological shortcomings are prevalent among studies of education, particularly in the selection of meaningful outcome measures. A summary of evidence-based considerations is provided for current and future program directors.


Novel DnaJ Protein Facilitates Thermotolerance of Transgenic Tomatoes.
DnaJ proteins, which are molecular chaperones that are widely present in plants, can respond to various environmental stresses. At present, the function of DnaJ proteins was studied in many plant species, but only a few studies were conducted in tomato. Here, we examined the functions of a novel tomato (Solanum lycopersicum) DnaJ protein (SlDnaJ20) in heat tolerance using sense and antisense transgenic tomatoes. Transient conversion assays of Arabidopsis protoplasts showed that SlDnaJ20 was targeted to chloroplasts. Expression analysis showed that SlDnaJ20 expression was induced by chilling, NaCl, polyethylene glycol, and H₂O₂, especially via heat stress. Under heat stress, sense plants showed higher fresh weights, chlorophyll content, fluorescence (Fv/Fm), and D1 protein levels, and a lower accumulation of reactive oxygen species (ROS) than antisense plants. These results suggest that SlDnaJ20 overexpression can reduce the photoinhibition of photosystem II (PSII) by relieving ROS accumulation. Moreover, higher expression levels of HsfA1 and HsfB1 were observed under heat stress in sense plants, indicating that SlDnaJ20 overexpression contributes to HSF expression. The yeast two-hybrid system proved that SlDnaJ20 can interact with the chloroplast heat-shock protein 70. Our results indicate that SlDnaJ20 overexpression enhances the thermotolerance of transgenic tomatoes, whereas suppression of SlDnaJ20 increases the heat sensitivity of transgenic tomatoes.


The Gentle Side of the UPS: Ubiquitin-Proteasome System and the Regulation of the Myogenic Program.
In recent years, the ubiquitin-proteasome system (UPS) has emerged as an important regulator of stem cell function. Here we review recent findings indicating that UPS also plays critical roles in the biology of satellite cells, the muscle stem cell responsible for its maintenance and regeneration. While we focus our attention on the control of key transcriptional regulators of satellite cell function, we briefly discuss early studies suggesting the UPS participates more broadly in the regulation of satellite cell stemness and regenerative capacity.


[A brief history of diagnosis and treatment of primary aldosteronism.].
Primary aldosteronism is a clinical syndrome characterized by hypokalemia, suppressed activities of plasma renin and high urinary and plasma aldosterone levels in hypertensive patients. It was first defined and reported by Jerome W. Conn in 1955. From 1960s to early 1970s, its techniques of diagnosis and treatment were greatly improved by the availability of spironolactone, realization of the renin-angiotensin-aldosterone system, and progress in laboratory tests and adrenal venous sampling. In 1970s, notwithstanding the extensive application of modern imaging modalities, such as CT scanning, adrenal venous sampling and steroid analysis have remained to be the most accurate and reliable localization method. From 1980s, more and more patients with primary aldosteronism were screened out from the hypertensive population by plasma renin activity/plasma aldosterone concentration ratio and cured by surgical interventions; laparoscopic unilateral adrenalectomy has become the generally accepted golden standard of operation.


Distribution and projections of neurons with immunoreactivity for both gastrin-releasing peptide and bombesin in the guinea-pig small intestine.
Bombesin-like and gastrin-releasing peptide (GRP)-like immunoreactivities were localized in nerves of the guinea-pig small intestine and celiac ganglion with the use of antibodies raised against the synthetic peptides. The anti-bombesin serum (preincubated to avoid cross reactivity with substance P) and the anti-GRP serum revealed the same population of neurons. Preincubation of the anti-bombesin serum with bombesin abolished the immunoreactivity in nerves while absorption of the anti-GRP serum with either bombesin or the 14-27 C-terminal of GRP only reduced the immunoreactivity. The immunoreactivity was abolished by incubation with GRP 1-27. Immunoreactive nerves were found in the myenteric plexus, circular muscle, submucous plexus and in the celiac ganglion. Faintly reactive nerve cell bodies were found in the myenteric ganglia (3.2% of all neurons) but not in submucous ganglia. After all ascending and descending pathways in the myenteric plexus had been cut, reactive terminals disappeared in the myenteric plexus, circular muscle (including the deep muscular plexus) and the submucous plexus on the anal side. After the mesenteric nerves were cut no changes were observed in the intestinal wall but the reactive fibres in celiac ganglia disappeared. It is deduced that GRP/bombesin-immunoreactive nerve cell bodies in myenteric ganglia project from the myenteric plexus to other myenteric ganglia situated further anally (average length 12 mm), anally to the circular muscle (average length 9 mm), anally to submucous ganglia (average length 13 mm) and external to the intestine to the celiac ganglia.(ABSTRACT TRUNCATED AT 250 WORDS)


Echocardiography derived three-dimensional printing of normal and abnormal mitral annuli.
The objective of this study was to assess the clinical feasibility of using echocardiographic data to generate three-dimensional models of normal and pathologic mitral valve annuli before and after repair procedures. High-resolution transesophageal echocardiographic data from five patients was analyzed to delineate and track the mitral annulus (MA) using Tom Tec Image-Arena software. Coordinates representing the annulus were imported into Solidworks software for constructing solid models. These solid models were converted to stereolithographic (STL) file format and three-dimensionally printed by a commercially available Maker Bot Replicator 2 three-dimensional printer. Total time from image acquisition to printing was approximately 30 min. Models created were highly reflective of known geometry, shape and size of normal and pathologic mitral annuli. Post-repair models also closely resembled shapes of the rings they were implanted with. Compared to echocardiographic images of annuli seen on a computer screen, physical models were able to convey clinical information more comprehensively, making them helpful in appreciating pathology, as well as post-repair changes. Three-dimensional printing of the MA is possible and clinically feasible using routinely obtained echocardiographic images. Given the short turn-around time and the lack of need for additional imaging, a technique we describe here has the potential for rapid integration into clinical practice to assist with surgical education, planning and decision-making.


Linear assembly of a porphyrin-C60 complex confined in vertical nanocylinders of amphiphilic block copolymer films.
Linear assemblies of a 1 : 1 porphyrin-fullerene C60 complex were formed in vertical cylindrical polyether nanodomains of amphiphilic block copolymer films by a simple spin coating-annealing method. The nanocylinder structures were retained even with high contents of the complex in the polymer films.


[Treatment of progressive, locally advanced prostate carcinoma].
Due to the lack of randomized and controlled trials comparing treatment options for locally advanced prostate cancer no clear cut therapeutical advise can be given. Currently, external beam radiotherapy is the form of treatment most often applied to which other therapeutic modalities have to be compared. Radical prostatectomy alone does not improve survival at this stage. The combination of local treatment modalities with neoadjuvant androgen deprivation seems to be a promising approach which has to be a promising approach which has to be evaluated in prospective clinical trials.


Baseline circulating tumour DNA and interim PET predict response in relapsed/refractory classical Hodgkin lymphoma.
Reliable biomarkers for early identification of treatment failure in relapsed/refractory (r/r) classical Hodgkin lymphoma (cHL) are lacking. Circulating tumour DNA (ctDNA) profiling has emerged as a powerful predictive and prognostic tool in several haemopoietic and non-haemopoietic malignancies and may guide rational treatment choices in r/r cHL. To assess the predictive and prognostic value of ctDNA, we performed a retrospective analysis on 55 r/r cHL patients treated with the bendamustine, gemcitabine and vinorelbine (BEGEV) regimen and additionally evaluated the potential utility of integrating ctDNA with interim [18 F]-FDG positron emission tomography (iPET). Baseline ctDNA genotyping in r/r cHL mirrored gene mutations and pathways involved in newly diagnosed cHL. We found that baseline ctDNA quantification and serial ctDNA monitoring have prognostic value in r/r cHL receiving salvage chemotherapy. Lastly, integrating ctDNA quantification with iPET evaluation may improve the early identification of patients at high risk of failing standard salvage therapy, who may benefit from an early switch to immunotherapeutic agents. Collectively, our results support the implementation of non-invasive methods to detect minimal residual disease in recurrent cHL and justify its prospective evaluation in appropriately designed clinical trials.


Tourniquets may increase postoperative swelling and pain after internal fixation of ankle fractures.
Tourniquets frequently are used in orthopaedic surgery to provide a bloodless field. There is still controversy among authors regarding the benefits and potential risks of using a tourniquet. The objective of this prospective randomized study was to quantify the effect of tourniquet use on postoperative swelling, pain, and range of motion after open reduction and internal fixation of ankle fractures. Fifty-four patients with closed ankle fractures were treated surgically; 26 patients were operated on using a thigh tourniquet (Group A), and 28 patients had surgery without the use of a tourniquet (Group B). The groups of patients were similar for age, gender, fracture types, and operative procedures. There was no difference in operation time. Using a tourniquet during open reduction and internal fixation of ankle fractures did increase postoperative swelling and postoperative pain at Day 5 and at Week 6 after surgery. We found a trend for a better range of motion in the ankle until the 6-week followup in the nontourniquet group compared with the group in which the tourniquet was inflated. According to these results we do not recommend using a tourniquet for osteosynthesis of ankle fractures. Therapeutic study, Level I-1 (randomized controlled trial). See the Guidelines for Authors for a complete description of levels of evidence.


Short-Term Genetic Selection for Adolescent Locomotor Sensitivity to Delta9-Tetrahydrocannabinol (THC).
Cannabis use is linked to positive and negative outcomes. Identifying genetic targets of susceptibility to the negative effects of cannabinoid use is of growing importance. The current study sought to complete short-term selective breeding for adolescent sensitivity and resistance to the locomotor effects of a single 10 mg/kg THC dose in the open field. Selection for THC-locomotor sensitivity was moderately heritable, with the greatest estimates of heritability seen in females from the F2 to S3 generations. Selection for locomotor sensitivity also resulted in increased anxiety-like activity in the open field. These results are the first to indicate that adolescent THC-locomotor sensitivity can be influenced via selective breeding. Development of lines with a genetic predisposition for THC-sensitivity or resistance to locomotor effects allow for investigation of risk factors, differences in consequences of THC use, identification of correlated behavioral responses, and detection of genetic targets that may contribute to heightened cannabinoid sensitivity.


Surgical methods for anorectal malformations from Rehbein to Peña--critical assessment of score systems and proposal for a new classification.
In this review article the surgical techniques of Rehbein, Romualdi, Kiesewetter, and Peña/De Vries are compared and corresponding surgical steps are pointed out. Peña/De Vries have adopted many surgical steps from Stephens and further standardized this technique. The results of recent postoperative examinations are presented with regard to the surgical methods of Stephens, Rehbein, Romualdi, Kiesewetter, and Peña. In that connection, the paper makes it clear that comparison of postoperative studies is not possible because the authors evaluate too small numbers of cases, use different scoring systems, do not indicate individual steps of their surgical techniques and do not consider the height of the atresias. The reference to the type of fistula that can be found in recent literature unfortunately does not provide a common basis for comparative studies either. Therefore, the paper suggests complete abandonment of scoring systems and classification of postoperative continence results instead according to the therapy that has to be applied. In addition, it again stresses the importance and necessity of adhering to the Wingspread classification.


The effects of horse-riding simulator exercise on balance in elderly with knee osteoarthritis.
[Purpose] The aim of this study was to examine the effects of horse-riding simulator exercise on balance in elderly with knee osteoarthritis. [Subjects and Methods] Fifty elderly patients with knee osteoarthritis were recruited, a horse-riding simulator group performed exercise three times a week for eight weeks. And each exercise was performed for 30 minutes. [Results] The horse-riding simulator group showed significant differences after the intervention in Short Form Berg Balance Scale, Functional reaching test. [Conclusion] The results of this study indicate that horse-riding simulator exercise is effective on knee osteoarthritis. Therefore, horse-riding simulator exercise can be used balance training for knee osteoarthritis.


Flap reconstruction following high pressure injection injuries of the hand: a report of three cases.
High pressure injection injuries are well known to cause significant injury to the hand, with high amputation rates and poor functional outcome. Surgical treatment consists of early aggressive debridement followed by secondary closure. Flap reconstruction is a reconstruction option which can increase the chance of digit salvage, as well as give an acceptable functional and cosmetic result. We review three cases of flap reconstruction following high pressure injection injuries, and discuss their role in the treatment of these injuries.


Corticotrophin-releasing factor type 2 receptor-mediated suppression of gonadotrophin-releasing hormone mRNA expression in GT1-7 cells.
Corticotrophin-releasing factor (CRF) released during stress has been implicated in the suppression of the hypothalamo-pituitary-gonadal (HPG) axis, especially the gonadotrophin-releasing hormone (GnRH) pulse generator, the central neural regulator of pituitary LH and FSH secretion, resulting in reproductive dysfunction. The gonadal steroid 17beta-oestradiol (E2) has been shown to enhance CRF- and stress-induced suppression of pulsatile LH secretion. In the present study, we investigated the potential direct action of CRF on GnRH neurones by using GT1-7 cells, an established GnRH cell line. Furthermore, we investigated the modulatory influence of E2 on the effects of CRF and expression of CRF type 2 receptors (CRF-R2). Expression of CRF-R2 in the GT1-7 cells was detected by reverse transcription-polymerase chain reaction (RT-PCR). CRF produced a dose-dependent suppression of GnRH mRNA expression, an effect reversed by the selective CRF-R2 antagonist, astressin2-B (Ast2-B). E2 combined with CRF resulted in a greater suppression of GnRH expression compared with either treatment alone. E2 also increased CRF-R2 expression. These results demonstrate for the first time expression of CRF-R2 in the GT1-7 cells and suggest that CRF may directly regulate GnRH gene expression, an effect mediated, at least in part, by CRF-R2. They also raise the possibility that up-regulation of CRF-R2 may contribute to the sensitising influence of E2 on CRF- and stress-induced suppression of the GnRH pulse generator.


Amended final report of the safety assessment of t-Butyl Alcohol as used in cosmetics.
t-Butyl Alcohol (t-BuOH) is a tertiary aliphatic alcohol that is used as a solvent or an alcohol denaturant and as a perfume carrier in cosmetics. t-BuOH was reported as an ingredient in 32 formulations of eye makeup, fragrance, and shaving preparations, at concentrations ranging from 0.00001% and 0.3%. There is little acute oral toxicity in animals; e.g., the acute oral LD(50) in rats was 3.0 to 3.7 g/kg. In short-term oral studies in rats, t-BuOH at 2% (w/v) or less in drinking water did not cause gross organ or tissue damage in mice, although weight loss was reported and microscopic damage to livers and kidney and alterations such as centrilobular necrosis, vacuolation in hepatocytes, and loss of hepatic architecture were noted. Subchronic oral dosing with t-BuOH increased the mineralization of the kidney, nephropathy, and urinary bladder transitional cell epithelial hyperplasia in rats; and liver damage, chronic inflammation, hyperplasia of transitional cell epithelium urinary, and proliferative changes including hyperplasia and neoplasia in the thyroid in mice. Male rats exposed to t-BuOH were susceptible to alpha 2mu-globulin nephropathy. t-BuOH (99.9%) was a moderate to severe ocular irritant to rabbits and caused mild to moderate dermal irritation to rabbits. It was not considered to be a primary dermal irritant to rabbits. In animal studies, fetotoxicity generally increased with concentration, and fetal weights were slightly depressed at concentrations of 0.5% to 1% t-BuOH. t-BuOH produced a significant increase in the number of resorptions per litter. There was also a significant decrease in the number of live fetuses per litter. t-BuOH reduced maternal weight gain, litter sizes, birth weights, and weights at weaning, and increased perinatal and postnatal mortality. t-BuOH was not mutagenic in several bacterial and mammalian test systems. The principal effects from 2 years of exposure to t-BuOH in drinking water (up to 10 mg/ml for rats and 20 mg/ml for mice) were proliferative lesions (hyperplasia, adenoma, and carcinoma) in the kidneys of exposed male rats, and nephropathy in all exposed groups of female rats. There was some evidence of carcinogenic activity, but it was not consistent between species, sexes, or doses. A repeat-insult patch test (RIPT) test showed no potential for eliciting either dermal irritation or sensitization by 100% t-BuOH. Dermatitis can result from dermal exposure of humans to t-BuOH. In consideration of these data, it was concluded that t-BuOH was (at most) a weak carcinogen and unlikely to have significant carcinogenic potential as currently used in cosmetic formulations. In addition, the renal tubule effects found in male rats were likely an effect of alpha 2mu-globulin. In consideration of the reproductive and developmental toxicity data, the increased incidence of still births occurred at high exposure levels and was likely secondary to maternal toxicity. Based on the available animal and clinical data in this report, it was concluded that t-BuOH is safe as used in cosmetic products.


Electrogenic ion transport in the mouse endometrium: functional aspects of the cultured epithelium.
A primary culture of mouse endometrial epithelium grown on permeable supports was established and the electrogenic ion transport across the endometrial epithelium was studied using the short-circuit current (I(SC)) technique. Enzymatically isolated mouse endometrial cells were immunostained with epithelial cells markers, cytokeratins, indicating an epithelial origin of the culture. Mouse endometrial epithelial cells grown on Millipore filters formed polarized monolayers with junctional complexes as revealed by light and electron microscopy. The cultured monolayers exhibited an average basal I(SC) of 4.6 +/- 0.3 microA/cm2, transepithelial voltage of 2.7 +/- 0.2 mV and transepithelial resistance of 599 +/- 30 omega cm2. The basal current was reduced by 85% in Na+-free solution and 13% in Cl(-)-free solution. The basal current could also be substantially (57.7%) blocked by an apical Na+ channel blocker, amiloride (10 microM), suggesting that Na+ absorption largely contributed to the basal current. Apical addition of Cl- channel blocker, DPC (2 mM), also exhibited an inhibitory effect, 19.4%, on the basal I(SC), indicating minor involvement of Cl- secretion as compared to that of Na+ absorption. The cultured endometrial epithelium also responded to a number of secretagogues including adrenaline and forskolin with increases in the I(SC), which could involve substantial Cl- secretion. The present study has established a culture of mouse endometrial epithelium exhibiting predominantly Na+ absorption under unstimulated condition, and Cl- secretion in response to various secretagogues. This culture may be useful for studying various regulatory mechanisms of electrogenic ion transport across the endometrial epithelium.


Challenges in perioperative blood transfusions in kidney transplantation and the need for Patient Blood Management.
Anemia is highly prevalent in end-stage chronic kidney disease patients, increasing their risk of receiving blood transfusions during and on the days after a kidney transplant (KTx) surgery. However, there is currently a lack of data that thoroughly describes this phenomenon in this population, the associated risk factors, and how they could benefit from the application of Patient Blood Management (PBM) guidelines. Observational study. All adult patients who received a KTx between January 1st, 2020, and December 31st, 2021, were included and followed up to six months after transplantation. Those who received a multiorgan transplant, whose data was missing in the electronic health records, and who had primary non-function were excluded. We recorded donor and recipient characteristics, cold ischemia time, preoperative hemoglobin concentration, iron status deficiency biomarkers, incidence of delayed graft function and biopsy-proven graft rejections, and graft function at discharge and 6 months after transplantation. We found that a high amount (39%) of KTx recipients required at least one blood transfusion during the perioperative period. And that 1) most of these patients had anemia at the time of transplantation (85.4%), 2) iron status upon admission was associated with the transfusion of more blood units (3.9 vs 2.7, p=0.019), 3) surgical reintervention (OR 7.28, 2.35-22.54) and deceased donor donation (OR 1.99, 1.24-3.21) were associated with an increased risk of transfusion, and finally, 4) there was an association between a higher number of blood units transfused and impaired kidney graft function six months after hospital discharge (1.6 vs 1.9, p=0.02). In conclusion, PBM guidelines should be applied to patients on the KTx deceased donor waiting list and especially those scheduled to receive a transplant from a living donor. This could potentially increase the utilization efficiency of blood products and avoid transfusion-related severe adverse effects.


Histopathologic study of partial hydatidiform moles and DNA triploid placentas.
Sixty-two placentas with a triploid DNA content, which were analyzed by flow cytometry using paraffin-embedded tissues, were histologically investigated. These placentas were histologically classified as follows: 51 partial hydatidiform moles (PM), two hydropic abortuses and nine non-hydropic placentas. The DNA indices of the triploid peaks were between 1.41 and 1.60. Histologically, two populations of normal and edematous villi, vesicular villous edema with cistern formation, focal syncytiotrophoblastic hyperplasia with vacuolation, and villous scalloping with trophoblastic inclusion were almost always observed in the PM, but no single pathologic feature was specific for PM. The two entities, PM and triploid placenta, overlapped. Not all triploid gestations are PM and not all PM moles are triploid as shown in previously reported diploid or tetraploid PM. Although no patient with triploid PM developed persistent disease in this series, follow up of triploid PM is required as long as its true biological potential remains unclear. Flow cytometry is a reliable aid in the diagnosis of PM.


Delayed accumulation (flip flop) of Tc-99m DTPA in a leiomyosarcoma.
An 85-year-old woman with leiomyosarcoma occupying the abdominopelvic area had a renal dynamic study with Tc-99m DTPA. The tumor area initially showed almost no content of the radiopharmaceutical. At 24 hours, there was retention of activity in the leiomyosarcoma as compared with surrounding tissues (flip flop). It was noted that, based on literature reports, tumors of mesodermal origin (leiomyoma, spindle cell sarcoma, neurofibroma) may have the ability to concentrate Tc-99m DTPA.


Divergent selection and phenotypic plasticity during incipient speciation in Lake Victoria cichlid fish.
Divergent selection acting on several different traits that cause multidimensional shifts are supposed to promote speciation, but the outcome of this process is highly dependent on the balance between the strength of selection vs. gene flow. Here, we studied a pair of sister species of Lake Victoria cichlids at a location where they hybridize and tested the hypothesis that divergent selection acting on several traits can maintain phenotypic differentiation despite gene flow. To explore the possible role of selection we tested for correlations between phenotypes and environment and compared phenotypic divergence (P(ST)) with that based on neutral markers (F(ST)). We found indications for disruptive selection acting on male breeding colour and divergent selection acting on several morphological traits. By performing common garden experiments we also separated the environmental and heritable components of divergence and found evidence for phenotypic plasticity in some morphological traits contributing to species differences.


[Development of degenerative changes in the wrist with pseudarthrosis of the scaphoid bone].
In patients with symptomatic scaphoid nonunion there was an increased evidence of progessive osteoarthrosis expressed as instability of the wrist; it is defined as a scapholunate angle more than 70 degrees or a radiolunate angle of more than 10 degrees. We tried to determine the factors of risk and prognostic indicators of degenerative arthritis. In this study 40 patients with painfull nonunion of the scaphoid were analysed. Duration of nonunion was not in correlation with development of osteoarthritic changes (p=0,644; p>0,05). Progessive degenerative changes correlated well with radiolunate angle (p=0,398; p<0,05), kapitolunate angle (p=0,381; p<0,05) and carpal index (p=0,392; p<0,05). The average values of intercarpal angles increased with progression of osteoarthritic changes. There was a statistically strong corelation between location of fracture in proximal third and presence of degenerative changes (p=0,341; p<0,01). Intesity of arthritic changes showed no statistically significant correlation regarding untreated fractures (p=0,665; p>0,05). We recommend that a scaphoid nonunion associated with carpal instability should be operated before degenerative changes develop.


Hydrocarbon-Stitched Peptide Agonists of Glucagon-Like Peptide-1 Receptor.
Glucagon-like peptide 1 (GLP-1) is a natural peptide agonist of the GLP-1 receptor (GLP-1R) found on pancreatic β-cells. Engagement of the receptor stimulates insulin release in a glucose-dependent fashion and increases β-cell mass, two ideal features for pharmacologic management of type 2 diabetes. Thus, intensive efforts have focused on developing GLP-1-based peptide agonists of GLP-1R for therapeutic application. A primary challenge has been the naturally short half-life of GLP-1 due to its rapid proteolytic degradation in vivo. Whereas mutagenesis and lipidation strategies have yielded clinical agents, we developed an alternative approach to preserving the structure and function of GLP-1 by all-hydrocarbon i, i + 7 stitching. This particular "stitch" is especially well-suited for reinforcing and protecting the structural fidelity of GLP-1. Lead constructs demonstrate striking proteolytic stability and potent biological activity in vivo. Thus, we report a facile approach to generating alternative GLP-1R agonists for glycemic control.


Genomic structure of the human prion protein gene.
Creutzfeld-Jacob disease and Gerstmann-Sträussler syndrome are rare degenerative disorders of the nervous system which have been genetically linked to the prion protein (PrP) gene. The PrP gene encodes a host glycoprotein of unknown function and is located on the short arm of chromosome 20, a region with few known genes or anonymous markers. The complete structure of the PrP gene in man has not been determined despite considerable interest in its relationship to these unusual disorders. We have determined that the human PrP gene has the same simple genomic structure seen in the hamster gene and consists of two exons and a single intron. In contrast to the hamster PrP gene the human gene appears to have a single major transcriptional start site. The region immediately 5' of the transcriptional start site of the human PrP gene demonstrates the GC-rich features commonly seen in housekeeping genes. Curiously, the genomic clone we have isolated contains a 24-bp deletion that removes one of five octameric peptide repeats predicted to form a B-pleated sheet in this region of the PrP. We have also identified 5' of the PrP gene an RFLP which has a high degree of heterozygosity and which should serve as a useful marker for the pter-12 region of human chromosome 20.


[Treatment of visual hallucinations with ramelteon in two patients with dementia Lewy bodies].
Herein, I report the improvement of visual hallucinations (VHs) in 2 patients with a clinical diagnosis of probable dementia with Lewy bodies (DLB) through the use of ramelteon, a selective MT1/MT2 melatonin receptor agonist. When first seen with complaints of VHs and insomnia in our memory clinic, their scores on the Mini-Mental State Examination were 25 and 20. One patient developed VHs at the age of 72 and could describe them in detail (e.g., a large crowd attending a midnight party). The other patient had dreamed of his deceased wife since the age of 78. He developed VHs at the age of 81 and could describe them in detail: his deceased wife was present and he tried to talk to her, but she did not respond. Both patients lived alone and telephoned their family with complaints of VHs in the middle of night, resulting in an increase in the burdens of care. Because cholinesterase inhibitors had been prescribed for both patients, further pharmacotherapeutic interventions targeted their insomnia rather than their VHs to avoid neuroleptic drug sensitivity, a recommendation of the DLB guidelines. Furthermore, ramelteon has few adverse effects. Within 8 weeks of initiating ramelteon, the VHs had disappeared and the insomnia had improved in both patients. Melatonin is recommended for the treatment of REM sleep behavior disorder (RBD), which is a frequent feature of DLB. The efficacy of ramelteon for VHs suggests a possible association of RBD with psychiatric symptoms in patients with DLB.