Endometrial inflammasome activation accompanies menstruation and may have implications for systemic inflammatory events of the menstrual cycle.
Does NLRP3 (NOD-, LRR- and pyrin domain-containing protein 3) inflammasome activation within decidualized endometrial stromal cells accompany menstruation and is this reflected systemically? Components of the NLRP3 inflammasome immunolocalize to decidualized endometrial stromal cells immediately prior to menstruation, and are activated in an in vitro model of menstruation, as evidenced by downstream interleukin (IL)-1beta and IL-18 release, this being reflected systemically in vivo. Menstruation is a highly inflammatory event associated with activation of NFκB (nuclear factor kappa-light-chain-enhancer of activated B cells), local release of chemokines and cytokines and inflammatory leukocyte influx. Systemically, chemokines and cytokines fluctuate across the menstrual cycle. This study examined the NLRP3 inflammasome and activation of downstream IL-1beta and IL-18 in endometrial tissues from women of known fertility (≥1 previous parous pregnancy) across the menstrual cycle (n ≥ 8 per cycle phase), serum from women during the proliferative, secretory and menstrual phases (≥9 per cycle phase) of the cycle and menstrual fluid collected on Day 2 of menses (n = 18). Endometrial stromal cells isolated from endometrial tissue biopsies (n = 10 in total) were used for an in vitro model of pre-menstrual hormone withdrawal. Expression and localization of components of the NLRP3 inflammasome (NLRP3 & apoptosis-associated speck-caspase recruit domain [ASC]) in endometrial tissues was performed by immunohistochemistry. Unbiased digital quantification of immunohistochemical staining allowed determination of different patterns of expression across the menstrual cycle. Serum from women across the menstrual cycle was examined for IL-1beta and IL-18 concentrations by ELISA. An in vitro model of hormone withdrawal from estrogen/progestin decidualized endometrial stromal cells was used to more carefully examine activation of the NLRP3 inflammasome. Endometrial stromal cells isolated from endometrial tissue biopsies (n = 10) were treated with estrogen/medroxyprogesterone acetate for 12 days to induce decidualization (assessed by release of prolactin) followed by withdrawal of steroid hormone support. Activation of NLRP3, & ASC in these cells was examined on Days 0-3 after hormone withdrawal by Western immunoblotting. Release of IL-1beta and IL-18 examined during decidualization and across the same time course of hormone withdrawal by ELISA. Specific involvement of NLRP3 inflammasome activation in IL-1beta and IL-18 release after hormone withdrawal was investigated via application of the NLRP3 inflammasome inhibitor MCC950 at the time of hormone withdrawal. Critical components of the NLRP3 inflammasome (NLRP3, ASC) were increased in menstrual phase endometrial tissues versus early secretory phase tissues (P < 0.05, n/s, respectively). NLRP3 and ASC were also elevated in the proliferative versus secretory phase of the cycle (P < 0.01, n/s, respectively) with ASC also significantly increased in the late-secretory versus early-secretory phase (P < 0.05). The pattern of activation was reflected in systemic levels of the inflammasome mediators, with IL-1beta and IL-18 elevated in peripheral blood serum during menstruation (Day 2 of menses) versus secretory phase (P = 0.026, P = 0.0042, respectively) and significantly elevated in menstrual fluid (Day 2 of menses) versus systemic levels across all cycle phases, suggesting that local inflammasome activation within the endometrium during menses is reflected by systemic inflammation. NLRP3 and ASC localized to decidualized cells adjacent to the spiral arterioles in the late secretory phase of the menstrual cycle, where the menstrual cascade is thought to be initiated, and to endometrial leukocytes during the menstrual phase. NLRP3 also localized to glandular epithelial cells during the late-secretory/menstrual phases. Localization of both NLRP3 and ASC switched from predominant epithelial localization during the early-secretory phase to stromal localization during the late-secretory/menstrual phase. Using an in vitro model of hormone withdrawal from decidualized human endometrial stromal cells, we demonstrated progressive activation of NLRP3 and ASC after hormone withdrawal increasing from Day 0 of withdrawal/Day 12 of decidualization to Day 3 of withdrawal. Downstream release of IL-1beta and IL-18 from decidualized stromal cells after hormone withdrawal followed the same pattern with the role of NLRP3 inflammasome activation confirmed via the inhibition of IL-1beta and IL-18 release upon application of MCC950. N/A. This study uses descriptive and semi-quantitative measures of NLRP3 inflammasome activation within endometrial tissues. Further, the in vitro model of pre-menstrual hormone withdrawal may not accurately recapitulate the in vivo environment as only one cell type is present and medroxyprogesterone acetate replaced natural progesterone due to its longer stability. We provide novel evidence that the NLRP3 inflammasome is activated within decidualized endometrial stromal cells immediately prior to menses and that local activation of the inflammasome within the endometrium appears to be reflected systemically in by activation of downstream IL-1beta and IL-18. Given the prevalence of menstrual disorders associated with inflammation including dysmenorrhoea and aspects of pre-menstrual syndrome, the inflammasome could be a novel target for ameliorating such burdens. The authors have no competing interests. J.E. was supported by a Fielding Foundation fellowship, NHMRC project grants (#1139489 and #1141946) and The Hudson Institute of Medical Research. L.A.S. was supported by The Hudson Institute of Medical Research and J.H. by an Australian Government Research Training Program Scholarship. We acknowledge the Victorian Government's Operating Infrastructure funding to the Hudson Institute. N/A.


Intrinsic functional connectivity in late-life depression: trajectories over the course of pharmacotherapy in remitters and non-remitters.
Previous studies in late-life depression (LLD) have found that patients have altered intrinsic functional connectivity in the dorsal default mode network (DMN) and executive control network (ECN). We aimed to detect connectivity differences across a treatment trial among LLD patients as a function of remission status. LLD patients (N=37) were enrolled into a 12-week trial of venlafaxine and underwent five functional magnetic resonance imaging resting state scans during treatment. Patients had no history of drug abuse, psychosis, dementia/neurodegenerative diseases or medical conditions with known effects on mood. We investigated whether there were differences in three networks: DMN, ECN and anterior salience network connectivity, as well as a whole brain centrality measure (eigenvector centrality). We found that remitters showed increases in ECN connectivity in the right precentral gyrus and decreases in DMN connectivity in the right inferior frontal gyrus and supramarginal gyrus. The ECN and DMN had regions (middle temporal gyrus and bilateral middle/inferior temporal/fusiform gyrus, respectively) that showed reversed effects (decreased ECN and increased DMN, respectively). Early changes in functional connectivity can occur after initial medication exposure. This study offers new data, indicating that functional connectivity changes differ depending on treatment response and can occur shortly after exposure to antidepressant medication.


Reliability of binocular vision measurements used in the classification of convergence insufficiency.
To evaluate the reliability of binocular vision measurements used in the classification of convergence insufficiency. Two examiners tested 20 fifth and sixth graders in a school setting who passed a screening of visual acuity, refraction, and binocularity. The tests, conducted using a standard protocol, consisted of von Graefe near heterophoria (NH), phorometric positive fusional vergence (PFV), nearpoint of convergence (NPC), and monocular pushup accommodative amplitude (AA). Each examiner measured each child three consecutive times for each test, on two separate occasions, spaced approximately 1 week apart. Intraexaminer and interexaminer agreement was assessed using intraclass correlation coefficients (ICC), the median absolute difference (MAD), and the coefficient of repeatability (COR). The within-session reliability of the NH (ICC: 0.95 to 0.99), NPC (ICC: 0.94 to 0.98), and AA (ICC: 0.88 to 0.95) were good, whereas the PFV was less reliable (ICC: 0.71 to 0.94). The intraexaminer reliability between sessions was good for the NPC (ICC: 0.92 and 0.89), less reliable for NH (ICC: 0.81 and 0.81) and AA (ICC: 0.89 and 0.69), and much less reliable for PFV break (ICC: 0.59 and 0.53). Typical between-session PFV differences (MAD) were between 3 and 4 delta, whereas the COR differences were as large as 12 delta. Three of the four measures (NH, NPC, and AA) often used in the classification of convergence insufficiency generally have good within-session and between-session reliability. The PFV break was found to have only fair reliability with clinically significant differences between sessions. The large potential test-retest differences found could complicate clinical decision-making in regards to diagnosis and treatment.


[Combination of 5-Fluorouracil and folinic acid--is it still the standard therapy for advanced colorectal carcinoma?].
After a long period in which the only therapeutic approach for the management of colorectal cancer was the optimization of fluoropyrimidine-based chemotherapy, the last few years have seen the emergence of newly active chemotherapeutic agents endowed with novel mechanisms of action, such as oxaliplatin (OHP), irinotecan (CPT11), raltitrexed and oral 5-fluorouracil (5-FU) pro-drugs which can offer new therapeutic possibilities. The availability of these different classes of cytotoxic agents has introduced the possibility of combination chemotherapy. The most widely studied combinations are 5-FU/folinic acid (FA)/CPT11 and 5-FU/FA/OHP, both of which have been tested in two randomized studies. These trials consistently demonstrated that the addition of CPT11 or OHP to widely accepted infusional/bolus 5-FU/FA regimens is superior to the same 5-FU/FA schedule alone in term of response rate (rate of about > 50%) and time to progression. It's worth nothing that both trials with the combination of 5-FU/FA/CPT11 showed a significant survival benefit. Toxicity profiles were more pronounced with the combination therapies but the quality of life evaluation showed that the combinations had no negative impact on the evolution of the global health status over time. A current ongoing randomized study compares 5-FU/FA/CPT11 with 5-FU/FA/OHP in order to identify the best first-line chemotherapy. Preliminary results in terms of objective response and toxicity are similar for both treatments. Raltitrexed is a thymidilate synthase inhibitor with activity comparable to 5-FU and a convenient administration schedule. In our institution, we performed a phase II single center trial to assess the efficacy of the combination of raltitrexed and CPT11. The principal aim of the study was the overall response rate. We observed a very promising 51% overall response rate. Most relevant side effects were diarrhea in about 20% of patients, asthenia and vomiting. Two recent studies have assessed the efficacy and tolerability of raltitrexed-OHP combination therapy. The results showed that the combination is active (ORR: 46-62%) and tolerable. Promising results have also been obtained in phase I studies with the OHP/CPT11 combination. All these data need to be confirmed in phase II-III clinical trials. In conclusion, combination therapy with CPT11 plus 5-FU/FA produces a 2-3 month survival advantage over 5-FU/FA alone and represents the new reference in the first-line chemotherapy of colorectal cancer. The association of 5-FU/FA and OHP improves response rates and progression-free survival. The increase in toxicity of these combinations is predictable and reversible and does not compromise quality of life. These important data suggest that there is now a limited role for single-agent first-line chemotherapy of metastatic colorectal cancer.


Results of the 2003 National Resident Matching Program: family practice.
The results of the 2003 National Resident Matching Program (NRMP) reflect a persistent decline of student interest in family practice residency training in the United States. Compared with the 2002 Match, 118 fewer positions (179 fewer US seniors) were filled in family practice residency programs through the NRMP in 2003, as well as 23 fewer (12 fewer US seniors) in primary care internal medicine, 20 fewer in pediatrics-primary care (11 fewer US seniors), and 23 fewer (34 fewer US seniors) in internal medicine-pediatric programs. In comparison, 40 more positions (14 more US seniors) were filled in anesthesiology and 8 more (8 more US seniors) in diagnostic radiology, two "marker" disciplines that have shown increases over the past 3 years. Sixty-seven more positions (but 148 fewer US seniors) were also filled in categorical internal medicine, while 107 more positions (33 more US seniors) were filled in categorical pediatrics programs, where trainees perceive options for either practicing as generalists or entering subspecialty fellowships, depending on the market. While the needs of the nation, especially rural and underserved populations, continue to offer opportunities for family physicians, family practice experienced continued decline though the 2003 NRMP. Many different forces, including student perspectives of the demands, rewards, and prestige of the specialty; the turbulence and uncertainty of the health care environment; liability protection issues; and the impact of faculty and resident role models, are impacting medical student career choices. The 2003 NRMP again confirms the trend away from family practice and primary care careers.


[The determination of the activity of natural killer cells in chronic lympholeukemia based on radiometric and morphological data].
Function of lymphocytes as effector cells of natural cytotoxicity in chronic lympholeukemia was investigated using radiometric and morphological methods. The effectiveness of these methods was compared. A significant decrease in NKC activity in B-cell type of chronic lympholeukemia was detected. A high degree of positive correlation between radiometry and morphology findings was revealed. The radionuclide (51Cr) method of investigation of NKC activity of lymphocytes was shown to have more advantages over the alternative morphological method in terms of practicability, accuracy and objectiveness.


Novel role of zinc in the regulation of prostate citrate metabolism and its implications in prostate cancer.
The prostate gland of humans and many other animals has the major function of accumulating and secreting extraordinarily high levels of citrate. This specialized metabolic process of "net citrate production" is the result of unique metabolic capabilities of the secretory epithelial cells. Most importantly, in prostate cancer (Pca) the capability for net citrate production is lost. In addition to citrate, the normal and BPH (benign prostatic hyperplasia) prostate also accumulates the highest levels of zinc in the body. As with citrate, in Pca the ability for high zinc accumulation is diminished. These and other correlations between zinc and citrate in the prostate have been indicative of an important role of zinc in the regulation of citrate metabolism in normal and malignant prostate epithelial cells. The link between zinc and citrate metabolism has now been established. The intramitochondrial accumulation of high zinc levels inhibits mitochondrial (m-) aconitase activity, which inhibits citrate oxidation. This essentially truncates the Krebs cycle and markedly decreases the cellular energy (ATP) production normally coupled to citrate oxidation. It is also clear that zinc accumulation in citrate-producing prostate epithelial cells is regulated by testosterone and by prolactin. These relationships form the basis for a new concept of the role of zinc and citrate-related energy metabolism in prostate malignancy. The inability of malignant prostate cells to accumulate high zinc levels results in increased citrate oxidation and the coupled ATP production essential for the progression of malignancy. The concept offers new approaches to the treatment of Pca.


Non-invasive In Vivo Imaging of Cancer Using Surface-Enhanced Spatially Offset Raman Spectroscopy (SESORS).
Rationale: The goal of imaging tumors at depth with high sensitivity and specificity represents a significant challenge in the field of biomedical optical imaging. 'Surface enhanced Raman scattering' (SERS) nanoparticles (NPs) have been employed as image contrast agents and can be used to specifically target cells in vivo. By tracking their unique "fingerprint" spectra, it becomes possible to determine their precise location. However, while the detection of SERS NPs is very sensitive and specific, conventional Raman spectroscopy imaging devices are limited in their inability to probe through tissue depths of more than a few millimetres, due to scattering and absorption of photons by biological tissues. Here, we combine the use of "Spatially Offset Raman spectroscopy" (SORS) with that of "surface-enhanced resonance Raman spectroscopy" (SERRS) in a technique known as "surface enhanced spatially offset resonance Raman spectroscopy" (SESO(R)RS) to image deep-seated glioblastoma multiforme (GBM) tumors in vivo in mice through the intact skull. Methods: A SORS imaging system was built in-house. Proof of concept SORS imaging was achieved using a PTFE-skull-tissue phantom. Imaging of GBMs in the RCAS-PDGF/N-tva transgenic mouse model was achieved through the use of gold nanostars functionalized with a resonant Raman reporter to create SERRS nanostars. These were then encapsulated in a thin silica shell and functionalized with a cyclic-RGDyK peptide to yield integrin-targeting SERRS nanostars. Non-invasive in vivo SORS image acquisition of the integrin-targeted nanostars was then performed in living mice under general anesthesia. Conventional non-SORS imaging was used as a direct comparison. Results: Using a low power density laser, GBMs were imaged via SESORRS in mice (n = 5) and confirmed using MRI and histopathology. The results demonstrate that via utilization of the SORS approach, it is possible to acquire clear and distinct Raman spectra from deep-seated GBMs in mice in vivo through the skull. SESORRS images generated using classical least squares outlined the tumors with high precision as confirmed via MRI and histology. Unlike SESORRS, conventional Raman imaging of the same areas did not provide a clear delineation of the tumor. Conclusion: To the best of our knowledge this is the first report of in vivo SESO(R)RS imaging. In a relevant brain tumor mouse model we demonstrate that this technique can overcome the limitations of conventional Raman imaging with regards to penetration depth. This work therefore represents a significant step forward in the potential clinical translation of SERRS nanoparticles for high precision cancer imaging.


Dual-Responsive Nanorobot-Based Marsupial Robotic System for Intracranial Cross-Scale Targeting Drug Delivery.
Nanorobots capable of active movement are an exciting technology for targeted therapeutic intervention. However, the extensive motion range and hindrance of the blood-brain barrier impeded their clinical translation in glioblastoma therapy. Here, a marsupial robotic system constructed by integrating chemical/magnetic hybrid nanorobots (child robots) with a miniature magnetic continuum robot (mother robot) for intracranial cross-scale targeting drug delivery is reported. For primary targeting on macroscale, the continuum robot enters the cranial cavity through a minimally invasive channel (e.g., Ommaya device) in the skull and transports the nanorobots to pathogenic regions. Upon circumventing the blood-brain barrier, the released nanorobots perform secondary targeting on microscale to further enhance the spatial resolution of drug delivery. In vitro experiments against primary glioblastoma cells derived from different patients are conducted for personalized treatment guidance. The operation feasibility within organisms is shown in ex vivo swine brain experiments. The biosafety of the treatment system is suggested in in vivo experiments. Owing to the hierarchical targeting method, the targeting rate, targeting accuracy, and treatment efficacy have improved greatly. The marsupial robotic system offers a novel intracranial local therapeutic strategy and constitutes a key milestone in the development of glioblastoma treatment platforms.


Stenting for left main coronary artery occlusion in adolescent: A case report.
Acute total or subtotal occlusion of left main coronary artery (LMCA) is a catastrophic and mostly fatal event. Patients may present with cardiogenic shock and die whenever this event occurs. Survival is strongly dependent on the presence of collateral blood flow to the left coronary artery or a dominant right coronary artery, and emergency intervention for preserving the left ventricular function. Here, we present a case of a 14-year-old boy with subtotal occlusion of the LMCA accompanying acute myocardial infarction probably caused by congenital syphilis according to his positive serum syphilis antibody. His survival was closely associated with a dominant right coronary artery and timely thrombolytic therapy. Finally, he was treated with angioplasty and paclitaxel-eluting stent implantation. He was followed up after stenting and was doing quite well at the time when we wrote this paper.


Research on prevalence of secondary conditions in individuals with disabilities: an overview.
Individuals with disabilities may be at increased risk of a number of secondary conditions. Secondary conditions influence health status and quality of life. Many of these conditions are preventable and their consequences can be managed. Knowledge of the prevalence, course, and association with age of secondary conditions is needed for better prevention and management. The aim of present study was to review the available literature on the prevalence of secondary conditions in individuals with disability. We performed searches of electronic databases for studies published between 1980 and 2017 that provided information on the prevalence of secondary conditions in individuals with disabilities. The reference list of all relevant citations was also reviewed for further material. Nineteen papers were included. The findings indicate that (i) studies were mainly population-based descriptive studies, (ii) researchers used different questionnaires, (iii) the most common studied population are patients with spinal cord injury, and (iv) even though secondary conditions are different in different patient populations, the main secondary conditions are pain, spasticity, urinary tract infections, fatigue, depression, and sleep problems. The prevalence of each condition varies widely depending on the studied population and research methodologies across studies. There is still much information that remains to be obtained on the prevalence, definition, and conceptual organization of secondary conditions. There is also a lack of longitudinal studies on the natural course of these conditions.


Studies on the cytotoxicity, antimicrobial and DNA-binding activities of plants used by the Ese'ejas.
Thirty-nine extracts of 13 plants used traditionally as medicinal by the Ese'ejas were studied in order to determine their cytotoxic effect in the brine shrimp. Infusions showed no toxicity. Those plants that tested positive for methanolic and dichloromethane extracts were assayed for DNA-binding activity. Cytotoxicity was not due to the presence of compounds that interact with DNA. Antimicrobial activity of plants used to treat infectious diseases was also performed for the decoctions. These proved to be active against some of the test microrganisms used in the assay.


Medical student reflections: Chaplain shadowing as a model for compassionate care training.
Training of compassionate and empathetic physicians requires commitment by educators to make it a priority. Chaplains typically have time and training to effectively demonstrate compassionate care in the clinical setting. This qualitative study aims to explore perceived benefits among medical students from pastoral care shadowing in integrating compassion and spirituality into education curricula. Sixty-four written reflections from first- and second-year medical students were collected from December 2018 to January 2020 after shadowing with hospital chaplains. Unprompted reflections were analyzed using coding networks. Four major themes identified included (1) learned values within pastoral care, (2) learned roles of pastoral care in the healthcare setting, (3) practiced spiritual assessment tools and resource identification, and (4) reflected personal impact on future career. Within each major theme, three to four sub-themes were further identified. Reflections support chaplain shadowing as a model for emphasizing spiritual and compassionate care through role-modeling, hands-on learning and reflective practices.


Reversal of neuromuscular block with a selective relaxant-binding agent: sugammadex.
Muscle relaxants are used in the perioperative period to aid in endotracheal intubation, facilitate surgical exposure, and in the critical care setting for prolonged relaxation. Until now, the only mechanism to reverse their effect is acetylcholinesterase inhibitors that result in excess parasympathetic activity and require a second drug to prevent this side effect. Additionally, the onset and degree of neuromuscular antagonism are often unpredictable and unreliable. Sugammadex is the first of the cyclodextrins to be used as a therapeutic agent. It quickly, effectively, and safely reverses steroidal neuromuscular blockers by encapsulating the muscle relaxant and rendering it inactive. Sugammadex may be considered the ideal reversal agent and the first drug in its class, which will likely change the practice of anesthesia and clinical neuromuscular pharmacology.


Silencing of the annexin II gene down-regulates the levels of S100A10, c-Myc, and plasmin and inhibits breast cancer cell proliferation and invasion.
To explore the roles of annexin II in breast cancer progression, and to study the effect of annexin II on breast cancer cell proliferation and invasion. This study was conducted in the Key Laboratory of Diagnostic Medicine Designated by the Ministry of Education, Chongqing Medical University, Chongqing, China from December 2006 to January 2009. First, we employed Western blot and reverse transcriptase polymerase chain reaction to detect the expression of annexin II and S100A10 in a panel of well-characterized human breast cancer cell lines, and investigated the localization of annexin II and S100A10 by use of immunofluorescence. We then silenced the expression of annexin II in MDA-MB-435s, which was found to over express annexin II, using the chemically-synthetic annexin II small interfering RNA (siRNA) duplexes (including 3 groups: blank MDA-MB-435s cells, cells transfected with negative control siRNA, and cells transfected with annexin II-siRNA). Finally, the cell proliferation, invasion, and plasmin generation were assayed, and the cellular levels of S100A10 and c-Myc were also detected. All the tests were repeated 3 times. Annexin II and S100A10 were over expressed in invasive human breast cancer cell lines. The siRNA targeting annexin II of MDA-MB-435s cells did not only decrease annexin II messenger RNA and protein levels, but also down-regulated the levels of S100A10, and c-Myc. The treated cells were remarkably blocked in the G0/G1 phase, and cells in the S/G2+M phase decreased. Additionally, the treatment with siRNA resulted in reduction of plasmin generation as well as a loss of the invasive capacity of breast cancer cells. Annexin II might be a key contributor to breast cancer proliferation and invasion.


Cystic fibrosis transmembrane conductance regulator recruitment to phagosomes in neutrophils.
Optimal microbicidal activity of human polymorphonuclear leukocytes (PMN) relies on the generation of toxic agents such as hypochlorous acid (HOCl) in phagosomes. HOCl formation requires H2O2 produced by the NADPH oxidase, myeloperoxidase derived from azurophilic granules, and chloride ion. Chloride transport from cytoplasm into phagosomes requires chloride channels which include cystic fibrosis transmembrane conductance regulator (CFTR), a cAMP-activated chloride channel. However, the phagosomal targeting of CFTR in PMN has not been defined. Using human peripheral blood PMN, we determined that 95-99% of lysosomal-associated membrane protein 1 (LAMP-1)-positive mature phagosomes were CFTR positive, as judged by immunostaining and flow cytometric analysis. To establish a model cell system to evaluate CFTR phagosomal recruitment, we stably expressed enhanced green fluorescent protein (EGFP) alone, EGFP-wt-CFTR and EGFP-DF508-CFTR fusion proteins in promyelocytic PLB-985 cells, respectively. After differentiation into neutrophil-like cells, CFTR presentation to phagosomes was examined. EGFP-wt-CFTR was observed to associate with phagosomes and colocalize with LAMP-1. Flow cytometric analysis of the isolated phagosomes indicated that such a phagosomal targeting was determined by the CFTR portion of the fusion protein. In contrast, significantly less EGFP-DF508-CFTR was found in phagosomes, indicating a defective targeting of the molecule to the organelle. Importantly, the CFTR corrector compound VRT-325 facilitated the recruitment of DF508-CFTR to phagosomes. These data demonstrate the possibility of pharmacologic correction of impaired recruitment of mutant CFTR, thereby providing a potential means to augment chloride supply to the phagosomes of PMN in patients with cystic fibrosis to enhance their microbicidal function.


Structure of the shiga-like toxin I B-pentamer complexed with an analogue of its receptor Gb3.
Shiga-like toxin I (SLT-I) is a virulence factor of Escherichia coli strains that cause disease in humans. Like other members of the Shiga toxin family, it consists of an enzymatic (A) subunit and five copies of a binding subunit (the B-pentamer). The B-pentamer binds to a specific glycolipid, globotriaosylceramide (Gb3), on the surface of target cells and thereby plays a crucial role in the entry of the toxin. Here we present the crystal structure at 2.8 A resolution of the SLT-I B-pentamer complexed with an analogue of the Gb3 trisaccharide. The structure reveals a surprising density of binding sites, with three trisaccharide molecules bound to each B-subunit monomer of 69 residues. All 15 trisaccharides bind to one side of the B-pentamer, providing further evidence that this side faces the cell membrane. The structural model is consistent with data from site-directed mutagenesis and binding of carbohydrate analogues, and allows the rational design of therapeutic Gb3 analogues that block the attachment of toxin to cells.


Effect of Stacking Sequence and Sodium Bicarbonate Treatment on Quasi-Static and Dynamic Mechanical Properties of Flax/Jute Epoxy-Based Composites.
The present paper deals with the investigation of quasi-static and dynamic mechanical response of epoxy-based composites reinforced with flax and/or jute plain weave fabrics. In order to evaluate the influence of the stacking sequence, two monolithic laminates reinforced with flax or jute fibers and two hybrid flax/jute laminates were manufactured through the vacuum infusion technique. Furthermore, an eco-friendly and cost-effective surface treatment based on fiber soaking in a sodium bicarbonate solution was employed to improve the fiber-matrix adhesion. The mechanical characterization (by means of quasi-static flexural, dynamic mechanical thermal analysis and Charpy impact tests) allowed to evidence that the sodium bicarbonate treatment leads to noticeable improvement of the mechanical performances of flax reinforced composites, whereas jute composites experience a slight decrease of their mechanical properties. Overall, the hybridization allows to achieve intermediate mechanical properties among those of monolithic composites. Furthermore, the coupled action of hybridization and surface treatment does not lead to a beneficial and reliable effect on the mechanical response of the resulting composites.


A theoretical study of 5,6,7,8-tetrahydro-6-hydroxymethylpterin: insight into intrinsic photoreceptor properties of 6-substituted tetrahydropterins.
Tetrahydropterins are essential biological cofactors, which play a crucial role in DNA and RNA syntheses, NO synthesis, hydroxylation of aromatic amino acids, etc. In the last few years, it has been shown that 6-substituted "unconjugated" tetrahydropterins can also play a photoreceptor chromophoric role in plants and cyanobacteria. However, the nature of the initial light signal transduction act in which H4pterins participate is unknown. Our quantum chemical calculations have shown the possibility of the fast internal conversion of excited states of H4pterins. The potential energy surface scan along the 1ππ* state shows no energy barrier leading to 1ππ*/S0 conical intersection, this explains the absence of fluorescence for H4pterins. Other trajectories of the internal conversion relate to the stretching vibrations of the N-H bonds of the pyrimidine ring for the Rydberg state. The presence of several trajectories of nonradiative quenching of the photoexcited singlet states provides the photostability of the molecule. It was demonstrated for the first time that the nature of the excited states of H4pterins is similar to the nature of the excited states of guanine.


Transient expression shows ligand gating and allosteric potentiation of GABAA receptor subunits.
Human gamma-aminobutyric acid A (GABAA) receptor subunits were expressed transiently in cultured mammalian cells. This expression system allows the simultaneous characterization of ligand-gated ion channels by electrophysiology and by pharmacology. Thus, coexpression of the alpha and beta subunits of the GABAA receptor generated GABA-gated chloride channels and binding sites for GABAA receptor ligands. Channels consisting of only alpha or beta subunits could also be detected. These homomeric channels formed with reduced efficiencies compared to the heteromeric receptors. Both of these homomeric GABA-responsive channels were potentiated by barbiturate, indicating that sites for both ligand-gating and allosteric potentiation are present on receptors assembled from either subunit.


Evolutionary dynamics of specialisation in herbivorous stick insects.
Understanding the evolutionary dynamics underlying herbivorous insect mega-diversity requires investigating the ability of insects to shift and adapt to different host plants. Feeding experiments with nine related stick insect species revealed that insects retain the ability to use ancestral host plants after shifting to novel hosts, with host plant shifts generating fundamental feeding niche expansions. These expansions were, however, not accompanied by expansions of the realised feeding niches, as species on novel hosts are generally ecologically specialised. For shifts from angiosperm to chemically challenging conifer hosts, generalist fundamental feeding niches even evolved jointly with strong host plant specialisation, indicating that host plant specialisation is not driven by constraints imposed by plant chemistry. By coupling analyses of plant chemical compounds, fundamental and ecological feeding niches in multiple insect species, we provide novel insights into the evolutionary dynamics of host range expansion and contraction in herbivorous insects.


Fold assignments for amino acid sequences of the CASP2 experiment.
New and newly extended methods for fold assignment were tested for their abilities to assign folds to amino acid target sequences of unknown three-dimensional structure. These target sequences, released through the CASP2 experiment, are not obviously related to any sequence of known three-dimensional (3D) structure. We assigned 3D folds to target sequences and filed these predictions with CASP2 before their 3D structures were released. The methods tested were (1) Environmental 3D profiles of Bowie and colleagues [Bowie, J.U., Luthy, R., Eisenberg, D. Science 253:164-170, 1991]; (2) A variation of this is termed Directional Profiles; (3) The H3P2 five-dimensional sequence-structure substitution matrix of Rice and Eisenberg [Rice, D., Eisenberg, D.J. Mol. Biol. 267:1026-1037, 1997]; and (4) The Sequence Derived Property methods of Fischer and Eisenberg [Fischer, D., Eisenberg, D. Prot. Sci. 5:947-955, 1996]. When the 3D structures of the sequences were released, 17 of our predictions were evaluated. Of these 17, we assigned high probabilities to 11, of which 9 were correct. Five of these correct predictions were of known 3D structures similar to the targets and four of these were of new folds. The evaluation demonstrated that our methods were effective in assigning the proper fold to more than half of the CASP2 targets with known folds (5/9) and also were able to detect half of the sequences that corresponded to no known folds (4/8). Even when the correct fold is assigned to a sequence, proper alignment of the sequence to the structure remains a challenge. Our methods were able to produce accurate alignments (< 1.2 mean residue shift error from the structural alignment) for four of the targets, including the particularly difficult alignment (only 7% residue identity in the structurally aligned regions) of the ferrochelatase sequence to the fold of a periplasmic binding protein.


Cortisol, 17 alpha-OH, progesterone and androgen responses to a standardized ACTH-stimulation in different stages of the normal menstrual cycle.
The release of cortisol, 17 alpha-OH-progesterone, androstenedione and testosterone during a standardized ACTH-stimulation test was investigated in three different stage of the normal menstrual cycle, to conclude if there is any stage dependency on the release of these hormones. No statistically significant differences were observed between the three stages concerning cortisol and testosterone increase. The increase of androstenedione in the pre-ovulatory stage was significantly high than that seen during the early follicular phase of the cycle. The increase of 17 alpha-OH-progesterone in the luteal phase was significantly less than that of both the early and late follicular stages of the cycle. Progesterone levels showed a small, but significant increase after ACTH-stimulation, in both the early and late stage of the follicular phase. However, the levels remained within the normal range of the follicular phase. In the luteal phase no increase was seen after ACTH-stimulation. Oestradiol-17 beta levels did not change at all after ACTH stimulation. The stage dependency of androstenedione and 17 alpha-OH-progesterone is discussed. The described stage-dependency different increase of 17 alpha-OH-progesterone release can be of importance when the results of ACTH-tests are evaluated to detect carriers of congenital adrenal hyperplasia.


Caregiver's Burden, Coping, and Psycho-Education in Indian Households with Single- and Multiple-Affected Members with Schizophrenia.
There is considerable evidence that family psycho-education combined with pharmacological intervention for patients with schizophrenia increases family understanding of the illness, reduces the familial burden of care, and improves patient outcomes. However, no studies have determined whether the burden of care is greater for those families with more than one ill member (multiplex) than for families with a single-affected individual (simplex), and whether psycho-educational programs should be adapted to meet the specific needs of multiplex families. This study was conducted at a tertiary care postgraduate teaching hospital in New Delhi, India. Caregivers in simplex [n = 50] and multiplex families [n = 30] were compared with regard to levels of burden, coping, and the impact of psycho-education on family functioning. All the caregiver participants attended eight bimonthly, psycho-educational intervention sessions. They were assessed on the Burden Assessment Schedule (BAS) and the Coping Check List (CCL) before and after psycho-education. Caregivers from the multiplex families reported significantly more burden on two domains of the BAS, but there were no significant differences between the groups with regard to coping on the CCL. Following psycho-education, significant improvement occurred in the majority of domains of the BAS and the CCL; the effect sizes varied by domain and family type. Multiplex families face a greater burden of care compared with simplex families. Currently, available psycho-education programs are moderately effective for such families.


Tracing prehistoric migrations by the viruses they carry: human T-cell lymphotropic viruses as markers of ethnic relationships.
Three reasons that HTLV-I and HTLV-II would not be expected to trace human migrations over extended time periods have been examined, and none has proven fatal to the theory. Transmission of the HTLVs (human T-cell lymphotropic viruses) in endemic settings highly depends on passage through breast milk, and this creates a pattern of distribution similar to that of mitochondrial DNA. The HTLVs probably evolve at variable rates, making the extent of sequence change a poor tool for dating human migrations. However, qualitative relationships between the sequence of human population separations and virus strain may be more regular. The uniqueness of viruses as markers of human relationship gives this method special value as a source of novel ideas regarding human movements and as independent confirmation of migration hypotheses that have been based on more conventional methods.


Opposed effects of enzymatic gliotoxin N- and S-methylations.
Gliotoxin (1), a virulence factor of the human pathogenic fungus Aspergillus fumigatus, is the prototype of epipoly(thiodioxopiperazine) (ETP) toxins. Here we report the discovery and functional analysis of two methyl transferases (MTs) that play crucial roles for ETP toxicity. Genome comparisons, knockouts, and in vitro enzyme studies identified a new S-adenosyl-l-methionine-dependent S-MT (TmtA) that is, surprisingly, encoded outside the gli gene cluster. We found that TmtA irreversibly inactivates ETP by S-alkylation and that this detoxification strategy appears to be not only limited to ETP producers. Furthermore, we unveiled that GliN functions as a freestanding amide N-MT. GliN-mediated amide methylation confers stability to ETP, damping the spontaneous formation of tri- and tetrasulfides. In addition, enzymatic N-alkylation constitutes the last step in gliotoxin biosynthesis and is a prerequisite for the cytotoxicity of the molecule. Thus, these specialized alkylating enzymes have dramatic and fully opposed effects: complete activation or inactivation of the toxin.


Size and position of a single condyle allograft influence knee kinematics.
An optimal match for size and shape between the donor femur and the host knee is considered a critical factor influencing the outcome of a knee allograft implantation. An in vitro allograft model was developed to determine the influence of the size and position of a lateral distal femoral condylar allograft on knee kinematics. Functional knee motion was simulated in a cadaver host knee in the intact state after removing and reimplanting the native lateral condyle of the distal femur and after serially replacing the native condyle with eight donor allografts. Each allograft was first tested in an optimal position and subsequently shifted 3 mm proximal and 3 mm distal to the joint line to quantify changes in joint kinematics due to the position of the allograft. The intact knee and the knee with the ideally implanted native allograft followed similar kinematic trends. Decreasing the width of the allograft increased the valgus knee orientation at full flexion, translated the tibia posteriorly at full extension, and externally rotated the tibia throughout knee flexion. The proximal shift in allograft position increased the valgus orientation at full extension, translated the tibia posteriorly at mid-flexion, and externally rotated the tibia throughout flexion. The distal shift in position had the opposite effect on the kinematics of the proximal shift. These results indicate that improving techniques for preoperative size-matching and intraoperative allograft placement may help to reduce biomechanical complications following implantation of the allograft.


Availability and costs of medicines for the treatment of tuberculosis in Europe.
To evaluate the access to comprehensive diagnostics and novel antituberculosis medicines in European countries. We investigated the access to genotypic and phenotypic Mycobacterium tuberculosis drug susceptibility testing and the availability of antituberculosis drugs and calculated the cost of drugs and treatment regimens at major tuberculosis treatment centres in countries of the WHO European region where rates of drug-resistant tuberculosis are the highest among all WHO regions. Results were stratified by middle-income and high-income countries. Overall, 43 treatment centres from 43 countries participated in the study. For WHO group A drugs, the frequency of countries with the availability of phenotypic drug susceptibility testing was as follows: (a) 75% (30/40) for levofloxacin, (b) 82% (33/40) for moxifloxacin, (c) 48% (19/40) for bedaquiline, and (d) 72% (29/40) for linezolid. Overall, of the 43 countries, 36 (84%) and 24 (56%) countries had access to bedaquiline and delamanid, respectively, whereas only 6 (14%) countries had access to rifapentine. The treatment of patients with extensively drug-resistant tuberculosis with a regimen including a carbapenem was available only in 17 (40%) of the 43 countries. The median cost of regimens for drug-susceptible tuberculosis, multidrug-resistant/rifampicin-resistant tuberculosis (shorter regimen, including bedaquiline for 6 months), and extensively drug-resistant tuberculosis (including bedaquiline, delamanid, and a carbapenem) were €44 (minimum-maximum, €15-152), €764 (minimum-maximum, €542-15152), and €8709 (minimum-maximum, €7965-11759) in middle-income countries (n = 12) and €280 (minimum-maximum, €78-1084), €29765 (minimum-maximum, €11116-40584), and €217591 (minimum-maximum, €82827-320146) in high-income countries (n = 29), respectively. In countries of the WHO European region, there is a widespread lack of drug susceptibility testing capacity to new and repurposed antituberculosis drugs, lack of access to essential medications in several countries, and a high cost for the treatment of drug-resistant tuberculosis.


Association of hyperglycemia and molecular subclass on survival in IDH-wildtype glioblastoma.
Hyperglycemia has been associated with worse survival in glioblastoma. Attempts to lower glucose yielded mixed responses which could be due to molecularly distinct GBM subclasses. Clinical, laboratory, and molecular data on 89 IDH-wt GBMs profiled by clinical next-generation sequencing and treated with Stupp protocol were reviewed. IDH-wt GBMs were sub-classified into RTK I (Proneural), RTK II (Classical) and Mesenchymal subtypes using whole-genome DNA methylation. Average glucose was calculated by time-weighting glucose measurements between diagnosis and last follow-up. Patients were stratified into three groups using average glucose: tertile one (<100 mg/dL), tertile two (100-115 mg/dL), and tertile three (>115 mg/dL). Comparison across glucose tertiles revealed no differences in performance status (KPS), dexamethasone dose, MGMT methylation, or methylation subclass. Overall survival (OS) was not affected by methylation subclass (P = .9) but decreased with higher glucose (P = .015). Higher glucose tertiles were associated with poorer OS among RTK I (P = .08) and mesenchymal tumors (P = .05), but not RTK II (P = .99). After controlling for age, KPS, dexamethasone, and MGMT status, glucose remained significantly associated with OS (aHR = 5.2, P = .02). Methylation clustering did not identify unique signatures associated with high or low glucose levels. Metabolomic analysis of 23 tumors showed minimal variation across metabolites without differences between molecular subclasses. Higher average glucose values were associated with poorer OS in RTKI and Mesenchymal IDH-wt GBM, but not RTKII. There were no discernible epigenetic or metabolomic differences between tumors in different glucose environments, suggesting a potential survival benefit to lowering systemic glucose in selected molecular subtypes.


Convergence of plant-rich and plant-only diets.
Discussants at the Third International Congress on Vegetarian Nutrition considered the nutritional adequacy, benefits, and health outcomes of plant-only (e.g., vegan and fruitarian), plant-based (e.g., macrobiotic, lactovegetarian, semivegetarian, and meatless), and omnivorous dietary patterns. The increased availability of a variety of plant foods, the advent of nutrient-fortified plant foods, the use of vitamin and mineral supplements, and the widespread dissemination of sound information on dietary patterns mean that convergence between the essential nutrient profiles of plant-only and plant-rich, plant-based diets is possible. Special attention should be paid to nutrition among vulnerable groups by age or physiologic status if they consume diets based solely on plants. Research has shown that both plant-only and plant-based eating patterns have health benefits, most notably in reducing the risk of chronic, degenerative diseases. The panel concluded that evidence for a convergence of scientific opinion on the safety and healthfulness of plant-only diets that are appropriately planned to meet all nutrient requirements compared with plant-based diets is considerable.


Activity and composition of ammonia oxidizing bacterial communities and emission dynamics of NH3 and N2O in a compost reactor treating organic household waste.
To monitor emissions of NH(3) and N(2)O during composting and link these to ammonia oxidation rates and the community structure of ammonia oxidizing bacteria (AOB). A laboratory-scale compost reactor treating organic household waste was run for 2 months. NH(3) emissions peaked when pH started to increase. Small amounts of N(2)O and CH(4) were also produced. In total, 16% and less than 1% of the initial N was lost as NH(3)-N and N(2)O-N respectively. The potential ammonia oxidation rate, determined by a chlorate inhibition assay, increased fourfold during the first 9 days and then remained high. Initially, both Nitrosospira and Nitrosomonas populations were detected using DGGE analysis of AOB specific 16S rRNA fragments. Only Nitrosomonas europaea was detected under thermophilic conditions, but Nitrosospira populations re-established during the cooling phase. Thermophilic conditions favoured high potential ammonia oxidation rates, suggesting that ammonia oxidation contributed to reduced NH(3) emissions. Small but significant amounts of N(2)O were emitted during the thermophilic phase. The significance of different AOBs detected in the compost for ammonia oxidation is not clear. This study shows that ammonia oxidation occurs at high temperature composting and therefore most likely reduces NH(3) emissions.


Beta-sitosterol from psyllium seed husk (Plantago ovata Forsk) restores gap junctional intercellular communication in Ha-ras transfected rat liver cells.
We purified compounds from the husks of psyllium seeds (Plantago ovata Forsk; desert Indian wheat), beginning with an ethanol extraction then followed by HP-20 and silica gel chromatography, which restored gap junctional intercellular communication (GJIC) in v-Ha-ras transfected rat liver epithelial WB-F344 cell line (WB-Ha-ras). GJIC was assessed by a scrape loading dye transfer assay. The active compound was identified as beta-sitosterol based on gas chromatography retention times and electron ionization mass spectroscopy (EI-MS) spectrum of authentic beta-sitosterol. Authentic beta-sitosterol restored GJIC in the tumorigenic WB-Ha-ras GJIC-deficient cells at a dose of 2.4 microM. In addition, a similar phytosterol, stigmasterol, also restored GJIC, albeit at a lower activity. beta-sitosterol and stigmasterol increased the level of connexin43 protein (Cx43) and restored phosphorylation of Cx43 to levels similar to the parental nontransfected cell line. We concluded that the restoration of intercellular communication in the GJIC-deficient, tumorigenic WB-Ha-ras cell line by the ethanol soluble fraction of psyllium seed husks is largely due to the presence of the phytosterol, beta-sitosterol. We discuss implications for dietary modulation of cancer by beta-sitosterol.


Midwives' experience of their education, knowledge and practice around immersion in water for labour or birth.
There is limited research examining midwives' education, knowledge and practice around immersion in water for labour or birth. Our aim was to address this gap in evidence and build knowledge around this important topic. This mixed method study was performed in two phases, between August and December 2016, in the birth centre of a tertiary public maternity hospital in Western Australia. Phase one utilised a cross sectional design to examine perceptions of education, knowledge and practice around immersion in water for labour or birth through a questionnaire. Phase two employed a qualitative descriptive design and focus groups to explore what midwives enjoyed about caring for women who labour or birth in water and the challenges midwives experienced with waterbirth. Frequency distributions were employed for quantitative data. Thematic analysis was undertaken to extract common themes from focus group transcripts. The majority (85%; 29 of 34) of midwives surveyed returned a questionnaire. Results from phase one confirmed that following training, 93% (27 of 29) of midwives felt equipped to facilitate waterbirth and the mean waterbirths required to facilitate confidence was seven. Midwives were confident caring for women in water during the first, second and third stage of labour and enjoyed facilitating water immersion for labour and birth. Finally, responses to labour and birth scenarios indicated midwives were practicing according to state-wide clinical guidance. Phase two included two focus groups of seven and five midwives. Exploration of what midwives enjoyed about caring for women who used water immersion revealed three themes: instinctive birthing; woman-centred atmosphere; and undisturbed space. Exploration of the challenges experienced with waterbirth revealed two themes: learning through reflection and facilities required to support waterbirth. This research contributes to the growing knowledge base examining midwives' education, knowledge and practice around immersion in water for labour or birth. It also highlights the importance of exploring what immersion in water for labour and birth offers midwives, as this research suggests they are integral to sustaining waterbirth as an option for low risk women.


A case-control study on red meat consumption and risk of stroke among a group of Iranian adults.
We aimed to examine the association between red meat consumption and stroke in a group of Iranian adults. A hospital-based case-control study. The study included stroke patients and hospital-based controls. Usual dietary intakes of participants were assessed by means of a validated 168-item semi-quantitative FFQ. Total red meat consumption was calculated by summing up the consumption of red, processed and visceral meats. One hundred and ninety-five cases were stroke patients hospitalized in the neurology ward and 195 controls were recruited from patients hospitalized in other wards with no history of cerebrovascular diseases or neurological disorders. Participants with stroke were older, more likely to be male and less likely to be obese. Individuals in the highest tertile of red meat intake were 119 % more likely to have stroke (OR=2·19; 95% CI 1·33, 3·60) compared with those in the lowest tertile. After controlling for age, sex and total energy intake, the association between red meat consumption and stroke was strengthened (OR=2·72; 95% CI 1·53, 4·83). This association remained significant even after further controlling for physical activity and smoking as well as dietary intakes. Additional adjustments for BMI, diabetes, hypertension and hyperlipidaemia did not influence the association significantly (OR=2·51; 95 % CI 1·19, 5·09). Consumption of red meat was associated with greater odds of having stroke in a group of Iranian adults.


Bovine sperm cells viability during incubation with or without exogenous DNA.
SummaryThe aim of this study was to assess the effect of exogenous DNA and incubation time on the viability of bovine sperm. Sperm were incubated at a concentration of 5 x 106/ml with or without plasmid pEYFP-NUC. Fluorescent probes, propidium iodide/Hoechst 33342, FITC-PSA and JC-1, were used to assess plasma membrane integrity (PMI), acrosome membrane integrity (AMI) and mitochondrial membrane potential (MMP) respectively at 0, 1, 2, 3 and 4 h of incubation. Exogenous DNA addition did not affect sperm viability; however, incubation time was related to sperm deterioration. Simultaneous assessment of PMI, AMI and MMP showed a reduction in the number of sperm with higher viability (integrity of plasma and acrosome membranes and high mitochondrial membrane potential) from 58.7% at 0 h to 7.5% after 4 h of incubation. Lower viability sperm (damaged plasma and acrosome membranes and low mitochondrial membrane potential) increased from 4.6% at 0 h to 25.9% after 4 h of incubation. When PMI, AMI and MMP were assessed separately we noticed a reduction in plasma and acrosome membrane integrity and mitochondrial membrane potential throughout the incubation period. Therefore, exogenous DNA addition does not affect sperm viability, but the viability is reduced by incubation time.


[THE MODES OF EVALUATION OF TYPE OF DEHYDRATION IN CHILDREN HOSPITALIZED BECAUSE OF ACUTE INTESTINAL INFECTION].
Every year about 800,000 cases of intestinal infections end in lethal outcome due to dehydration. The different types of dehydration acquire differential approach to correction. Everywhere there is no application of routine detection of osmolarity of blood plasma under exicosis in children in view of absence of possibility of instrumental measurement. The search of techniques is needed to make it possible to indirectly detect types of dehydration in children hospitalized because of acute intestinal infection with purpose to apply rationale therapy of water-electrolyte disorders. The sampling of 32 patients with intestinal infections accompanied with signs of exicosis degree I-III was examined. The detection of osmolarity of blood was implemented by instrumental technique using gas analyzer ABL 800 Flex (Radiometer; Denmark) and five estimate techniques according to results of biochemical analysis of blood. The differences in precision of measurement of osmolarity of blood plasma by instrumental and estimate techniques were compared using Bland-Altman graphic technique. It is established that formula: 2x[Na+kp] + [glucosekp] (mmol/l) is the most recise. Its application provided results comparable with values detected by instrumental mode.


Attacks on Ukrainian healthcare facilities during the first year of the full-scale Russian invasion of Ukraine.
Although the Geneva Conventions and Rome Statute demand protections for healthcare facilities during war, breaches of these protections are frequently reported. The ongoing war in Ukraine is no exception, with several healthcare attacks eliciting widespread condemnation. The Ukrainian Healthcare Center (UHC) has been collecting, verifying and documenting attacks on health infrastructure since the Russia-Ukraine War was launched. The aim of this study was to assess UHC documented healthcare facility attacks during the first year (24 February 2022 to 25 February 2023) of the full-scale Russian invasion of Ukraine. The Berkeley Protocol on Digital Open Source Investigations was used to document healthcare attacks. Data collection included temporal factors, location, facility type, attack and weapon type, number of killed and injured healthcare personnel and civilians, and whether facilities were damaged, destroyed or attacked more than once. There were 334 documented attacks on 267 Ukrainian healthcare facilities, with 230 facilities being damaged and 37 destroyed. General hospitals, primary care clinics, emergency departments and children's hospitals were most frequently targeted. The majority of attacks took place during the first three months and in eastern Ukrainian oblasts. Heavy weaponry was employed in almost all attacks. The total number of casualties included 97 fatalities and 114 injuries. During the first year of the full-scale Russian invasion of Ukraine, there were 334 attacks on 267 Ukrainian healthcare facilities documented by the UHC. Heavy weaponry was commonly used, and the direct impact of attacks was considerable in terms of facility damage and casualty tolls.


In-vivo administration of CLC-K kidney chloride channels inhibitors increases water diuresis in rats: a new drug target for hypertension?
The human kidney-specific chloride channels ClC-Ka (rodent ClC-K1) and ClC-Kb (rodent ClC-K2) are important determinants of renal function, participating to urine concentration and blood pressure regulation mechanisms. Here we tested the hypothesis that these chloride channels could represent new drug targets for inducing diuretic and antihypertensive effects. To this purpose, the CLC-K blockers benzofuran derivatives MT-189 and RT-93 (10, 50, 100 mg/kg), were acutely administered by gavage in Wistar rats, and pharmacodynamic and pharmacokinetic parameters determined by functional, bioanalytical, biochemical and molecular biology assays. Plasma concentration values for MT-189 and RT-93 were indicative of good bioavailability. Both MT-189 and RT-93 dose-dependently increased urine volume without affecting electrolyte balance. A comparable reduction of SBP was observed in rats after MT-189, RT-93 or furosemide administration. Benzofuran derivatives treatment did not affect kidney CLC-K mRNA level or inner medulla osmolality, whereas a significant vasopressin-independent down-regulation of aquaporin water channel type 2 was observed at protein and transcriptional levels. In rats treated with benzofuran derivatives, the observed polyuria was mainly water diuresis; this finding indirectly supports a cross-talk between chloride and water transport in nephron. Moreover, preliminary in-vitro evaluation of the drugs capability to cross the blood-inner ear barrier suggests that these compounds have a limited ability to induce potential auditory side effects. CLC-K blockers may represent a new class of drugs for the treatment of conditions associated with expanded extracellular volume, with a hopeful high therapeutic potential for hypertensive patients carrying ClC-K gain-of-function polymorphisms.


[Malignant transformation of mouse bone marrow cells induced in vitro by 60Co gamma-ray irradiation].
Mouse bone marrow cells were irradiated in vitro with 300, 200 and 100 rad of 60Co gamma-ray. Morphological transformation ensued from day 42 after exposure, but only in cells receiving 300 rad. Malignant transformation appeared in the 2nd generation in the subcultures. Some biological characteristics of malignant transformation were observed as follows: The attaching rate was 40.0%. Malignant transformation rate was 0.10-0.45%; Some colonies and clusters were formed in the semi-solid agar medium; There was no cell proliferation or colony formation in anti-ouabain inhibition test which implies that no mutation took place in the malignant transformation cells; Cytochalasin B rendered the cells to form a large number of multinuclear cells; Obvious chromosome aberration in number and structure was observed; Subsequent induction of fibrosarcomas in sites of subcutaneous inoculation of the malignant transformed cells in immunosuppressed mice. The results show that mouse bone marrow mesenchymal cells can undergo malignant transformation by large dose of gamma-ray irradiation in vitro. Criteria of malignant transformation: some clusters of cells formed in the semi-solid agar medium, multinuclear cell formation by action of cytochalasin B, and tumor mass induced in the location of inoculation.


[Applications of videolaparoscopic surgery in children]
OBJECTIVES: To present the videolaparoscopies performed by the Childreńs Surgery Service, and study the main indications for pediatric laparoscopic surgeries, considering advantages and disadvantages over conventional open procedures. METHODS: Retrospective analysis of 612 videolaparoscopies in children aged between 8 days and 17 years treated from November/95 to 2000. Basic principles of videolaparoscopy and the postoperative management of several pediatric diseases are described. The results, advantages, and complications were analyzed after a 5-year follow-up period. RESULTS: Laparoscopic surgery allowed for a wide series of abdominal procedures conventionally carried out through open surgery, mainly for the treatment of gastroesophageal reflux disease, inflammatory acute abdomen, adhesive intestinal obstruction, biliary lithiasis, tumors, cryptorchidism, ovarian diseases, splenectomies, aganglionosis, trauma and others. Morbidity was low (1%), and mortality due to laparoscopy was nonexistent. Conversion to laparotomy occurred in only 14 cases (2.3%), mainly because of trauma. The principal advantages included minimal surgical trauma, pain and reflex ileum, short hospital stay, almost no incisional hernias and better cosmetic scars. CONCLUSIONS: Videolaparoscopy seems to be a great advance in modern pediatric surgery, allowing safer and less invasive treatment of a wide series of abdominal diseases at all pediatric ages.


High-resolution X-ray structure of the unexpectedly stable dimer of the [Lys(-2)-Arg(-1)-des(17-21)]endothelin-1 peptide.
Previous structural studies on the [Lys((-2))-Arg((-1))]endothelin-1 peptide (KR-ET-1), 540-fold less potent than ET-1, strongly suggested the presence of an intramolecular Arg(-1)-Asp(8) (R(-1)-D(8)) salt bridge that was also observed in the shorter [Lys((-2))-Arg((-1))-des(17-21)]endothelin-1 derivative (KR-CSH-ET). In addition, for these two analogues, we have shown that the Lys-Arg dipeptide, which belongs to the prosequence, significantly improves the formation of the native disulfide bonds (>or=96% instead of approximately 70% for ET-1). In contrast to what was inferred from NMR data, molecular dynamics simulations suggested that such an intramolecular salt bridge would be unstable. The KR-CSH-ET peptide has now been crystallized at pH 5.0 and its high-resolution structure determined ab initio at 1.13 A using direct methods. Unexpectedly, KR-CSH-ET was shown to be a head-to-tail symmetric dimer, and the overall interface involves two intermolecular R(-1)-D(8) salt bridges, a two-stranded antiparallel beta-sheet, and hydrophobic contacts. Molecular dynamics simulations carried out on this dimer clearly showed that the two intermolecular salt bridges were in this case very stable. Sedimentation equilibrium experiments unambiguously confirmed that KR-ET-1 and KR-CSH-ET also exist as dimers in solution at pH 5.0. On the basis of the new dimeric structure, previous NMR data were reinterpreted. Structure calculations were performed using 484 intramolecular and 38 intermolecular NMR-derived constraints. The solution and the X-ray structures of the dimer are very similar (mean rmsd of 0.85 A). Since the KR dipeptide at the N-terminus of KR-CSH-ET is present in the prosequence, it can be hypothesized that similar intermolecular salt bridges could be involved in the in vivo formation of the native disulfide bonds of ET-1. Therefore, it appears to be likely that the prosequence does assist the ET-1 folding in a chaperone-like manner before successive cleavages that yield the bioactive ET-1 hormone.


A localized region of basal body proliferation in growing cells of Dileptus visscheri (ciliata, gymnostomate).
In the ciliate Dileptus organisms are completely organized at the time of excystation. They grow intensively, increasing in volume several times, before they reach the trophic cell size. The growth includes both the somatic part of the body as well as the oral parts, consisting of the cytostomal field and the ventral band of the proboscis. From the time of excystation, during the growth of the cell, the oral apparatus is able to capture and ingest food. Growth of an oral apparatus while functioning has not previously been reported in cilates. The proliferation of new oral kinetosomes in the growing oral apparatus occurs only in the cytostomal field. There is no proliferation of ciliature in the oral kineties of the growing proboscis. In those regions of the somatic kineties bordering the cytostomal field an extensive proliferation of somatic ciliature takes place. The zone of proliferation common to the oral and somatic ciliature supplies new kinetosomes for the elongating proboscis, the growing oral apparatus and the growth of the rest of the body. The mode of proliferation of the oral kinetosomes differs from that of the somatic kinetosomes. The oral kinetosomes form randomly in large numbers, appearing similar to an anarchic field such as is found in other ciliates. The somatic kinetosomes always arise singly and in a defined position in relation to the mature kinetosome. The proliferation of basal bodies in a defined area in Dileptus suggests the existence of a growth zone localized around the cytostomal field.


Pharmacological Mechanism of NRICM101 for COVID-19 Treatments by Combined Network Pharmacology and Pharmacodynamics.
Symptom treatments for Coronavirus disease 2019 (COVID-19) infection and Long COVID are one of the most critical issues of the pandemic era. In light of the lack of standardized medications for treating COVID-19 symptoms, traditional Chinese medicine (TCM) has emerged as a potentially viable strategy based on numerous studies and clinical manifestations. Taiwan Chingguan Yihau (NRICM101), a TCM designed based on a medicinal formula with a long history of almost 500 years, has demonstrated its antiviral properties through clinical studies, yet the pharmacogenomic knowledge for this formula remains unclear. The molecular mechanism of NRICM101 was systematically analyzed by using exploratory bioinformatics and pharmacodynamics (PD) approaches. Results showed that there were 434 common interactions found between NRICM101 and COVID-19 related genes/proteins. For the network pharmacology of the NRICM101, the 434 common interacting genes/proteins had the highest associations with the interleukin (IL)-17 signaling pathway in the Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Moreover, the tumor necrosis factor (TNF) was found to have the highest association with the 30 most frequently curated NRICM101 chemicals. Disease analyses also revealed that the most relevant diseases with COVID-19 infections were pathology, followed by cancer, digestive system disease, and cardiovascular disease. The 30 most frequently curated human genes and 2 microRNAs identified in this study could also be used as molecular biomarkers or therapeutic options for COVID-19 treatments. In addition, dose-response profiles of NRICM101 doses and IL-6 or TNF-α expressions in cell cultures of murine alveolar macrophages were constructed to provide pharmacodynamic (PD) information of NRICM101. The prevalent use of NRICM101 for standardized treatments to attenuate common residual syndromes or chronic sequelae of COVID-19 were also revealed for post-pandemic future.


Random forest classifiers trained on simulated data enable accurate short read-based genotyping of structural variants in the alpha globin region at Chr16p13.3.
In regions where reads don't align well to a reference, it is generally difficult to characterize structural variation using short read sequencing. Here, we utilize machine learning classifiers and short sequence reads to genotype structural variants in the alpha globin locus on chromosome 16, a medically-relevant region that is challenging to genotype in individuals. Using models trained only with simulated data, we accurately genotype two hard-to-distinguish deletions in two separate human cohorts. Furthermore, population allele frequencies produced by our methods across a wide set of ancestries agree more closely with previously-determined frequencies than those obtained using currently available genotyping software.


Food, mechanic and septic complications in patients enterally nutritioned in home conditions.
Home enteral nutrition (HEN for short) allows practically normal living for patients who cannot be fed orally but at the same time do not have to stay in hospitals, which is often found to decrease their mental condition, increase of probability of complications and costs of medical treatment. The aim of the study was to analyze the frequency of nutritional, mechanical and septic complications in patients fed enterally in home conditions. The study performed using retrospective analysis of study results and reports from control visits for patients in the period between 2012-2013. 147 patients fed enterally using HEN method participated in the study, including 70 men and 77 women aged 19 to 99 years (average 65 years). The following type of gastrointestinal tract access was used for patients: PEG in 113 (76.5%), feeding jejunostomy - 21 (1.4%), PEG-PEJ - 5 (3.5%), in case of the remaining 8 patients the nasogastric gavage (5.5%) was used. The most common complication were infections (of gastric tract, skin soft tissue in the region of nutritional fistula entry, in three cases the aspiration pneumonia was diagnosed) found in 55 (49.1%) of cases. Mechanical complications were found out in 29 (25.9% of all complications), nutritional complications were present 28 times, which constituted 25% of all complications. In the studied group of patients with an implemented HEN procedure, septic complications were the most common problem. The longest average nutrition time with PEG-PEJ probably results from the effective protection of the patient against aspiration pneumonia.


Molecular basis and characteristics of KATP channel in human corporal smooth muscle cells.
Relaxation of the corpus cavernosum smooth muscle is an absolute prerequisite for penile erection. Potassium channels play a role in the physiologic regulation of corporal smooth muscle tone. In spite of the physiological importance of K(ATP) channel in the modulation of corporal smooth muscle tone, there is a shortage of information available about the K(ATP) channel subtype(s) present in the corporal smooth muscle. The purpose of this study was to investigate the subunit type of K(ATP) channel, that is, the combinations of the Kir subunit and the SUR subunit in the human corporal smooth muscle and determine whether the electrophysiological kinetics and pharmacological properties of K(ATP) channels meet the subunit characteristics of the ion channel. We used cultured human corporal smooth muscle cells. To determine the presence of Kir and SURs subunits, RT-PCR was performed using Kir6.1, Kir6.2, SUR1, SUR2A, and SUR2B gene-specific primers. For electrophysiological recordings, the whole-cell, inside-out, and cell-attached configurations of the patch-clamp technique were used. We observed transcripts for Kir6.1, Kir6.2, and SUR2B in mRNA isolated from smooth muscle cells of cultured human corpus carvernosum. We recorded the unitary K(ATP) channel under the condition of intracellular and extracellular 140 mM [K(+)], and the slope conductance of the channel was 42.0+/-2.6 pS which is an intermediate conductance between that of either Kir6.1 or Kir6.2. The pinacidil (10 microM) increased the magnitude of the outward K(+) current (214.6+/-89.2%, n=12, < or = 0.05), which was blocked by the subsequent addition of the specific K(ATP) channel subtype selective blocker, glibenclamide (10 microM). The SIN-1(200 microM) induced increases in whole-cell outward K(+) currents (126.0+/-1.4%, n=4). The increased currents by SIN-1 were inhibited by glibenclamide (10 microM). We are the first to show that K(ATP) channel in human corporal smooth muscle is composed of Kir6.1-Kir6.2 construct expressed with SUR2B by RT-PCR. These findings, taken together with the electrophysiological results, suggest that K(ATP) channel in corporal smooth muscle cells is composed of heteromultimers of Kir6.1 and Kir6.2 with the ratio of 3 : 1 or 4 : 0 and SUR2B.


Electron mobility and injection dynamics in mesoporous ZnO, SnO₂, and TiO₂ films used in dye-sensitized solar cells.
High-performance dye-sensitized solar cells are usually fabricated using nanostructured TiO(2) as a thin-film electron-collecting material. However, alternative metal-oxides are currently being explored that may offer advantages through ease of processing, higher electron mobility, or interface band energetics. We present here a comparative study of electron mobility and injection dynamics in thin films of TiO(2), ZnO, and SnO(2) nanoparticles sensitized with Z907 ruthenium dye. Using time-resolved terahertz photoconductivity measurements, we show that, for ZnO and SnO(2) nanoporous films, electron injection from the sensitizer has substantial slow components lasting over tens to hundreds of picoseconds, while for TiO(2), the process is predominantly concluded within a few picoseconds. These results correlate well with the overall electron injection efficiencies we determine from photovoltaic cells fabricated from identical nanoporous films, suggesting that such slow components limit the overall photocurrent generated by the solar cell. We conclude that these injection dynamics are not substantially influenced by bulk energy level offsets but rather by the local environment of the dye-nanoparticle interface that is governed by dye binding modes and densities of states available for injection, both of which may vary from site to site. In addition, we have extracted the electron mobility in the three nanoporous metal-oxide films at early time after excitation from terahertz conductivity measurements and compared these with the time-averaged, long-range mobility determined for devices based on identical films. Comparison with established values for single-crystal Hall mobilities of the three materials shows that, while electron mobility values for nanoporous TiO(2) films are approaching theoretical maximum values, both early time, short distance and interparticle electron mobility in nanoporous ZnO or SnO(2) films offer considerable scope for improvement.


Thermosensitive TRPV channel subunits coassemble into heteromeric channels with intermediate conductance and gating properties.
Heat-sensitive transient receptor potential (TRP) channels (TRPV1-4) form the major cellular sensors for detecting temperature increases. Homomeric channels formed by thermosensitive TRPV subunits exhibit distinct temperature thresholds. While these subunits do share significant sequence similarity, whether they can coassemble into heteromeric channels has been controversial. In the present study we investigated the coassembly of TRPV subunits using both spectroscopy-based fluorescence resonance energy transfer (FRET) and single-channel recordings. Fluorescent protein-tagged TRPV subunits were coexpressed in HEK 293 cells; FRET between different subunits was measured as an indication of the formation of heteromeric channels. We observed strong FRET when fluorescence signals were collected selectively from the plasma membrane using a "spectra FRET" approach but much weaker or no FRET from intracellular fluorescence. In addition, no FRET was detected when TRPV subunits were coexpressed with members of the TRPM subfamily or CLC-0 chloride channel subunits. These results indicate that a substantial fraction of TRP channels in the plasma membrane of cotransfected cells were heteromeric. Single-channel recordings confirmed the existence of multiple heteromeric channel forms. Interestingly, heteromeric TRPV channels exhibit intermediate conductance levels and gating kinetic properties. As these subunits coexpress both in sensory neurons and in other tissues, including heart and brain, coassembly between TRPV subunits may contribute to greater functional diversity.


Hedonic eating, obesity, and addiction result from increased neuropeptide Y in the nucleus accumbens during human brain evolution.
The nucleus accumbens (NAc) is central to motivation and action, exhibiting one of the highest densities of neuropeptide Y (NPY) in the brain. Within the NAc, NPY plays a role in reward and is involved in emotional behavior and in increasing alcohol and drug addiction and fat intake. Here, we examined NPY innervation and neurons of the NAc in humans and other anthropoid primates in order to determine whether there are differences among these various species that would correspond to behavioral or life history variables. We quantified NPY-immunoreactive axons and neurons in the NAc of 13 primate species, including humans, great apes, and monkeys. Our data show that the human brain is unique among primates in having denser NPY innervation within the NAc, as measured by axon length density to neuron density, even after accounting for brain size. Combined with our previous finding of increased dopaminergic innervation in the same region, our results suggest that the neurochemical profile of the human NAc appears to have rendered our species uniquely susceptible to neurophysiological conditions such as addiction. The increase in NPY specific to the NAc may represent an adaptation that favors fat intake and contributes to an increased vulnerability to eating disorders, obesity, as well as alcohol and drug dependence. Along with our findings for dopamine, these deeply rooted structural attributes of the human brain are likely to have emerged early in the human clade, laying the groundwork for later brain expansion and the development of cognitive and behavioral specializations.


Mass identification of chloroplast proteins of endosymbiont origin by phylogenetic profiling based on organism-optimized homologous protein groups.
Chloroplasts originate from ancient cyanobacteria-like endosymbiont. Several tens of chloroplast proteins are encoded by the chloroplast genome, while more than hundreds are encoded by the nuclear genome in plants and algae, but the exact number and identity of nuclear-encoded chloroplast proteins are still unknown. We describe here attempts to identify a large number of unidentified chloroplast proteins of endosymbiont origin (CPRENDOs). Our strategy consists of whole genome protein clustering by the homolog group method, which is optimized for organism number, and phylogenetic profiling that extract groups conserved in cyanobacteria and photosynthetic eukaryotes. An initial minimal set of CPRENDOs was predicted without targeting prediction and experimentally validated.


Looking for the mechanism of arsenate respiration of Fusibacter sp. strain 3D3, independent of ArrAB.
The literature has reported the isolation of arsenate-dependent growing microorganisms which lack a canonical homolog for respiratory arsenate reductase, ArrAB. We recently isolated an arsenate-dependent growing bacterium from volcanic arsenic-bearing environments in Northern Chile, Fusibacter sp. strain 3D3 (Fas) and studied the arsenic metabolism in this Gram-positive isolate. Features of Fas deduced from genome analysis and comparative analysis with other arsenate-reducing microorganisms revealed the lack of ArrAB coding genes and the occurrence of two arsC genes encoding for putative cytoplasmic arsenate reductases named ArsC-1 and ArsC-2. Interestingly, ArsC-1 and ArsC-2 belong to the thioredoxin-coupled family (because of the redox-active disulfide protein used as reductant), but they conferred differential arsenate resistance to the E. coli WC3110 ΔarsC strain. PCR experiments confirmed the absence of arrAB genes and results obtained using uncouplers revealed that Fas growth is linked to the proton gradient. In addition, Fas harbors ferredoxin-NAD+ oxidoreductase (Rnf) and electron transfer flavoprotein (etf) coding genes. These are key molecular markers of a recently discovered flavin-based electron bifurcation mechanism involved in energy conservation, mainly in anaerobic metabolisms regulated by the cellular redox state and mostly associated with cytoplasmic enzyme complexes. At least three electron-bifurcating flavoenzyme complexes were evidenced in Fas, some of them shared in conserved genomic regions by other members of the Fusibacter genus. These physiological and genomic findings permit us to hypothesize the existence of an uncharacterized arsenate-dependent growth metabolism regulated by the cellular redox state in the Fusibacter genus.


Explaining the variation in the attained power of a stepped-wedge trial with unequal cluster sizes.
In a cross-sectional stepped-wedge trial with unequal cluster sizes, attained power in the trial depends on the realized allocation of the clusters. This attained power may differ from the expected power calculated using standard formulae by averaging the attained powers over all allocations the randomization algorithm can generate. We investigated the effect of design factors and allocation characteristics on attained power and developed models to predict attained power based on allocation characteristics. Based on data simulated and analyzed using linear mixed-effects models, we evaluated the distribution of attained powers under different scenarios with varying intraclass correlation coefficient (ICC) of the responses, coefficient of variation (CV) of the cluster sizes, number of cluster-size groups, distributions of group sizes, and number of clusters. We explored the relationship between attained power and two allocation characteristics: the individual-level correlation between treatment status and time period, and the absolute treatment group imbalance. When computational time was excessive due to a scenario having a large number of possible allocations, we developed regression models to predict attained power using the treatment-vs-time period correlation and absolute treatment group imbalance as predictors. The risk of attained power falling more than 5% below the expected or nominal power decreased as the ICC or number of clusters increased and as the CV decreased. Attained power was strongly affected by the treatment-vs-time period correlation. The absolute treatment group imbalance had much less impact on attained power. The attained power for any allocation was predicted accurately using a logistic regression model with the treatment-vs-time period correlation and the absolute treatment group imbalance as predictors. In a stepped-wedge trial with unequal cluster sizes, the risk that randomization yields an allocation with inadequate attained power depends on the ICC, the CV of the cluster sizes, and number of clusters. To reduce the computational burden of simulating attained power for allocations, the attained power can be predicted via regression modeling. Trial designers can reduce the risk of low attained power by restricting the randomization algorithm to avoid allocations with large treatment-vs-time period correlations.


Regulation of ribosomal RNA and 5s RNA synthesis in Drosophila melanogaster. I. Bobbed mutants.
Analysis of the rates and amounts of rRNA and 5s RNA synthesized in Drosophila melanogaster bobbed mutants was done by using acrylamide-gel electrophoresis. The results show that the amounts of rRNA synthesized are constant, although the rates of rRNA synthesis in bb's are reduced to 30% of the wild-type level. The rates of synthesis of 5s RNA were constant. The rate of synthesis of the two kinds of molecules that enter in equimolar amounts into the mature ribosome is non-coordinated.-The rates of rRNA synthesis were shown to be proportional to the length of the scutellar bristles, supporting the notion that in trichogen cells there is no developmental delay, but the size of the bristle depends directly on the rate of rRNA synthesis.


Case-Control Study of Individuals with Discrepant Nucleocapsid and Spike Protein SARS-CoV-2 IgG Results.
Laboratory-based methods for SARS-CoV-2 antibody detection vary widely in performance. However, there are limited prospectively-collected data on assay performance, and minimal clinical information to guide interpretation of discrepant results. Over a 2-week period, 1080 consecutive plasma samples submitted for clinical SARS-CoV-2 IgG testing were tested in parallel for anti-nucleocapsid IgG (anti-N, Abbott) and anti-spike IgG (anti-S1, EUROIMMUN). Chart review was conducted for samples testing positive or borderline on either assay, and for an age/sex-matched cohort of samples negative by both assays. CDC surveillance case definitions were used to determine clinical sensitivity/specificity and conduct receiver operating characteristics curve analysis. There were 52 samples positive by both methods, 2 positive for anti-N only, 34 positive for anti-S1 only, and 27 borderline for anti-S1. Of the 34 individuals positive for anti-S1 alone, 8 (24%) had confirmed COVID-19. No anti-S1 borderline cases were positive for anti-N or had confirmed/probable COVID-19. The anti-N assay was less sensitive (84.2% [95% CI 72.1-92.5%] vs 94.7% [95% CI 85.4-98.9%]) but more specific (99.2% [95% CI 95.5-100%] vs 86.9% [95% CI 79.6-92.3%]) than anti-S1. Abbott anti-N sensitivity could be improved to 96.5% with minimal effect on specificity if the index threshold was lowered from 1.4 to 0.6. Real-world concordance between different serologic assays may be lower than previously described in retrospective studies. These findings have implications for the interpretation of SARS-CoV-2 IgG results, especially with the advent of spike antigen-targeted vaccination, as a subset of patients with true infection are anti-N negative and anti-S1 positive.


Bioavailability of the polyphenols: status and controversies.
The current interest in polyphenols has been driven primarily by epidemiological studies. However, to establish conclusive evidence for the effectiveness of dietary polyphenols in disease prevention, it is useful to better define the bioavailability of the polyphenols, so that their biological activity can be evaluated. The bioavailability appears to differ greatly among the various phenolic compounds, and the most abundant ones in our diet are not necessarily those that have the best bioavailability profile. In the present review, we focus on the factors influencing the bioavailability of the polyphenols. Moreover, a critical overview on the difficulties and the controversies of the studies on the bioavailability is discussed.


Comprehensive proteomic profiling of adult Angiostrongylus costaricensis, a human parasitic nematode.
Angiostrongylus costaricensis is a nematode helminth that causes an intestinal acute inflammatory process known as abdominal angiostrongyliasis, which is a poorly understood human disease occurring in Latin America. Our aim was to study the proteomic profiles of adult parasites focusing on immunogenic proteins. Total cellular extracts from both genders showed similar 2-DE profiles, with 60% of all protein spots focused between pH 5-7 and presenting molecular masses from 20.1 to 66 kDa. A total of 53 different dominant proteins were identified in our dataset and were mainly associated with the following over-represented Gene Ontology Biological Process terms: "macromolecule metabolic process", "developmental process", "response to stress", and "biological regulation". Female and male immunoblots showed similar patterns of reactive proteins. Immunoreactive spots identified by MALDI-PSD were found to represent heat shock proteins, a putative abnormal DAuer Formation family member, and galectins. To date, very few biochemical analyses have focused on the nematode Angiostrongylus costaricensis. As such, our results contribute to a better understanding of its biology and the mechanisms underlying the host-parasite relationship associated with this species. Moreover, our findings represent a first step in the search for candidate proteins for diagnostic assays and the treatment of this parasitic infection.


Congenital heart disease and genetic syndromes: new insights into molecular mechanisms.
Advances in genetics allowed a better definition of the role of specific genetic background in the etiology of syndromic congenital heart defects (CHDs). The identification of a number of disease genes responsible for different syndromes have led to the identification of several transcriptional regulators and signaling transducers and modulators that are critical for heart morphogenesis. Understanding the genetic background of syndromic CHDs allowed a better characterization of the genetic basis of non-syndromic CHDs. In this sense, the well-known association of typical CHDs in Down syndrome, 22q11.2 microdeletion and Noonan syndrome represent paradigms as chromosomal aneuploidy, chromosomal microdeletion and intragenic mutation, respectively. Area covered: For each syndrome the anatomical features, distinctive cardiac phenotype and molecular mechanisms are discussed. Moreover, the authors include recent genetic findings that may shed light on some aspects of still unclear molecular mechanisms of these syndromes. Expert commentary: Further investigations are needed to enhance the translational approach in the field of genetics of CHDs. When there is a well-established definition of genotype-phenotype (reverse medicine) and genotype-prognosis (predictive and personalized medicine) correlations, hopefully preventive medicine will make its way in this field. Subsequently a reduction will be achieved in the morbidity and mortality of children with CHDs.


Optical properties of silver nanowire arrays with 35 nm periodicity.
We present highly ordered Ag nanowire arrays with 35nm periodicity grown on patterned templates. The optical properties measured using generalized ellipsometry exhibit strong anisotropy. Dielectric functions are calculated by fitting the Jones matrix elements with a biaxial layer model, accounting for both metallic behavior and localized surface plasmon resonances. The amplitude and wavelength maximum of the plasmon resonance perpendicular to the wires increase with increasing wire width and thickness. The dielectric coefficients of 10-mm-wide nanowires show a transition behavior from insulating in UV to metallic above 550nm. Their potential application as polarization-dependent plasmonic-scattering transparent conductive electrodes is discussed.


Quantitation of scanning electron microscopic urinary cytology.
Using scanning electron microscopy (SEM), differences in cell surface morphology are identifiable between normal urothelium and malignant urothelia including the presence of pleomorphic microvilli (PMV). PMV have been reported in carcinomas of the urinary bladder, and they appear early in the pathogenesis of these tumors in animal models. Preliminary studies in our laboratory and others' demonstrated similar changes in tissue and cytologic specimens from patients with bladder cancer. We observed and evaluated normal and neoplastic cells of the human bladder by SEM in a preliminary assessment of surface topography with regard to tumor growth, grade, and stage. However, while these SEM observations indicated that differences between normal and abnormal urothelia could be readily recognized, the distinction between the various types of atypical cells was not as clearly defined. Data collected so far indicates that changes in the surface topography of the exfoliated cells may possibly vary both in relation to tumor grade and stage. Based on these qualitative observations, we expanded our approach by using a computerized image analysis system directly interfaced with the SEM. The measurements which can be made include cell surface area, diameter, length, width, perimeter, orientation and number of PMV per unit area. Statistical analysis is also performed. The boundaries between cells are not recognizable by the system, making the single cells present in cytologic specimens ideal for evaluation. Uniform short microvilli are readily distinguished from PMV. Preliminary evaluation of 23 patients has distinguished specimens from malignant cases compared to patients with benign lesions.


Physiological and Psychological Effects of Volatile Organic Compounds from Dried Common Rush (Juncus effusus L. var. decipiens Buchen.) on Humans.
This study compared the participants' physiological responses and subjective evaluations of air scented with different concentrations of common rush (Juncus effusus L. var. decipiens Buchen.) (30 g and 15 g, with fresh air as a control). We asked 20 participants to complete a series of visual discrimination tasks while inhaling two different air samples. We evaluated (1) brain activity, (2) autonomic nervous activity, and (3) blood pressure and pulse rate, (4) in combination with self-evaluation. In addition, we quantified the concentrations of volatile organic compounds. The participants reported the scent to be sour, pungent, and smelly; this impression was likely caused by hexanal and acetic acid. Although the self-evaluations showed that participants did not enjoy the scent, their alpha amplitudes of electroencephalogram and parasympathetic nervous activity were increased, suggesting that participants were relaxed in this atmosphere. Moreover, a lower concentration resulted in a greater induction of relaxation. While the air was not pleasant-smelling, the volatile organic compounds present had a positive psychophysiological impact.