HIV counseling and testing in a tertiary care hospital in Ganjam district, Odisha, India.
Human immunodeficiency virus (HIV) counseling and testing (HCT) conducted at integrated counseling and testing centers (ICTCs) is an entry point, cost-effective intervention in preventing transmission of HIV. To study the prevalence of HIV among ICTC attendees, sociodemographic characteristics, and risk behaviors of HIV-seropositive clients. It was hospital record-based cross-sectional study of 26,518 registered ICTC clients at a tertiary care hospital in Ganjam district, Odisha, India over a 4-year period from January 2009 to September 2012. A total of 1732 (7.5%) out of 22,897 who were tested for HIV were seropositive. Among HIV-seropositives, 1138 (65.7%) were males, while 594 (34.3%) were females. Majority (88.3%) of seropositives were between the age group of 15-49 years. Client-initiated HIV testing (12.1%) was more seropositive compared to provider-initiated (2.9%). Among discordant couples, majority (95.5%) were male partner/husband positive and female partner/wife negative. Positives were more amongst married, less educated, low socioeconomic status, and outmigrants (P<0.0001). Risk factors included heterosexual promiscuous (89.3%), parent-to-child transmission 5.8%, unknown 3.1%, infected blood transfusion 0.8%, homosexual 0.5%, and infected needles (0.5%). There is need to encourage activities that promote HCT in all health facilities. This will increase the diagnosis of new HIV cases. The data generated in ICTC provide an important clue to understand the epidemiology in a particular geographic region and local planning for care and treatment of those infected with HIV and preventive strategies for those at risk especially married, young adults, and outmigrants to reduce new infections.


Cytotoxicity and antibacterial activity of scutellarein and carajurone-enriched fraction obtained from the hydroethanolic extract of the leaves of Fridericia chica (Bonpl.) L.G. Lohmann.
The hydroethanolic extract obtained from the dry leaves of Fridericia chica (HEFc) underwent several fractionations by different chromatographic techniques. The ethyl acetate and dichloromethane fraction were subjected to phytochemical analysis, resulting in the identification and isolation of scutellarein (1) and in a fraction rich in carajurone (2). They were tested for cytotoxicity in CHO-K1 and the antibacterial activity and mode of action by in vitro assays. The HEFc and scutellarein (1) presented no cytotoxicity. The results showed good antibacterial effect of HEFc against Streptococcus pyogenes and Staphylococcus aureus and moderate activity for Staphylococcus epidermidis, Pseudomonas aeruginosa and Salmonella typhimurium. The fraction containing the compound carajurone (2) showed good activity against Staphylococcus aureus and Staphylococcus epidermidis and moderate activity against Streptococcus pyogenes. Scutellarein (1) showed no activity against the bacteria tested. HEFc antibacterial mode of action appeared to be associated with changes in the permeability of bacterial membranes and nucleotide leakage.


[The use of titanium mesh plates for graft fixation in mandibular osteoplasty].
Opportunities of the use of mesh titanium plates as fixing devices are considered at reconstruction operations. Data of following up 8 patients with defects of the mandible of various location and extent are analysed. They were operated with reconstruction of the mandible with iliac graft and bone graft fixing with the mesh titanium plates. A technique of the operations is described, a case of successful use of mesh titanium implant modelled by the form of the mandible in repair of the mandible defect. The analysis of long-term results shows the potential efficiency of the use of mesh titanium plates as fixing adaptations in bone plastic operations.


Overexpression of GRIM-19 in cancer cells suppresses STAT3-mediated signal transduction and cancer growth.
Constitutive activation of signal transducer and activator of transcription 3 (STAT3) is common in many human and murine cancer cells, and its activation leads to cellular transformation. STAT3 pathway inhibitors have been reported to suppress cancer growth. To investigate the antitumor effects of inhibiting the STAT3-mediated signaling cascade in the cancer microenvironment, using a molecular-targeting approach, we focused on the gene associated with retinoid-IFN-induced mortality 19 (GRIM-19). GRIM-19 has been reported to interact physically with STAT3 and inhibit STAT3-dependent signal transduction. We used the nona-arginine (R9)-protein transduction domain (R9-PTD) as a protein carrier to induce high levels of GRIM-19 expression in vitro and in vivo. We generated an R9-PTD-containing GRIM-19 fusion protein (rR9-GRIM19) and successfully induced overexpression in the cytoplasm of cancer cells. Analysis of the expression of downstream molecules of STAT3 confirmed that in vitro rR9-GRIM19 treatment of constitutively activated STAT3 (STAT3c) cancer cells significantly reduced STAT3-dependent transcription. Moreover, intratumoral injections of rR9-GRIM19 in STAT3c cancer-bearing mice significantly suppressed tumor growth. These results suggest that intratumoral injections of rR9-GRIM19 have potential as a novel anticancer therapy in STAT3c cancer due to their ability to inhibit STAT3-mediated signal transduction without major systemic side effects.


Child abuse reporting by physicians.
Despite the prominent role of physicians in the child abuse reporting statutes of all 50 states, there is little published information concerning child abuse reporting by physicians. Our study examines the number and type of child abuse reports originating from physicians compared to other reporting sources. Reports by physicians comprised only 8% of the total number of child abuse reports received in Virginia in 1983. Reports by physicians were founded more often than reports from any other source, however, so that physicians contributed 14% of the reports that were ultimately founded. When compared to reports by all other sources, physician reports contained a significantly greater proportion of cases of physical abuse (P less than .0001), but a smaller proportion of physical neglect (P less than .0001). Physicians appear to be reporting those types of abuse that they are in a unique position to observe.


[Functional organ preservation in laryngeal and hypopharyngeal cancer].
The principles of open vs. laser microsurgical approaches for partial resections of the larynx are described, oncological as well as functional results discussed and corresponding outcomes following primary radiotherapy are opposed. Over the last decade, the endoscopic partial resection of the larynx has developed to an accepted approach in the treatment of early glottic and supraglottic carcinomas thus leading to a remarkable decline in the use of open surgery. Comparing the various surgical approaches of laryngeal partial resections, the oncological outcome of the patients, as far as survival and organ preservation are concerned, are comparable, whereas functional results of the endoscopic procedures are superior with less morbidity. The surgical procedures put together, are all superior to radiotherapy concerning organ preservation. Transoral laser microsurgery has been used successfully for vocal cord carcinomas with impaired mobility or fixation of the vocal cord, supraglottic carcinomas with infiltration of the pre- and/or paraglottic space as well as for selected hypopharyngeal carcinomas. It has been well documented that laser microsurgery achieves good oncological as well as functional results with reasonable morbidity. However, patients with those tumours have been successfully treated by open partial resections of the larynx at medical centres with appropriate expertise. The initially enthusiastic assessment of study results concerning the efficacy of various protocols of chemoradiation with the intent of organ preservation for laryngeal and hypopharyngeal carcinomas are judged more cautious, today, due to recent reports of rather high rates of late toxicity complications.


The CSC proteins FAP61 and FAP251 build the basal substructures of radial spoke 3 in cilia.
Dynein motors and regulatory complexes repeat every 96 nm along the length of motile cilia. Each repeat contains three radial spokes, RS1, RS2, and RS3, which transduct signals between the central microtubules and dynein arms. Each radial spoke has a distinct structure, but little is known about the mechanisms of assembly and function of the individual radial spokes. In Chlamydomonas, calmodulin and spoke-associated complex (CSC) is composed of FAP61, FAP91, and FAP251 and has been linked to the base of RS2 and RS3. We show that in Tetrahymena, loss of either FAP61 or FAP251 reduces cell swimming and affects the ciliary waveform and that RS3 is either missing or incomplete, whereas RS1 and RS2 are unaffected. Specifically, FAP251-null cilia lack an arch-like density at the RS3 base, whereas FAP61-null cilia lack an adjacent portion of the RS3 stem region. This suggests that the CSC proteins are crucial for stable and functional assembly of RS3 and that RS3 and the CSC are important for ciliary motility.


Caffeic acid phenethyl ester guards against benign prostate hypertrophy in rats: Role of IGF-1R/protein kinase-B (Akt)/β-catenin signaling.
Benign prostate hypertrophy (BPH) is among the most common diseases with a huge impact on the quality of life of elderly men. There is a current need for the development of well-tolerated and effective preventive strategies to improve the clinical outcome. Caffeic acid phenethyl ester (CAPE) is an important active ingredient isolated from honey-bee propolis with potent anti-proliferative, anti-inflammatory and antioxidant effects. These properties promote CAPE as a promising candidate to be tested as an alternative therapy for BPH, which is still uninvestigated. Herein, we tested the ability of CAPE to guard against testosterone-induced BPH and investigated the involvement of IGF1-R/Akt/β-catenin signaling as a protective mechanism in testosterone-induced BPH rat model. Treatment with CAPE reduced testosterone-induced increase in the prostate index and histopathological alterations. In addition, co-treatment with CAPE significantly suppressed insulin-like growth factor-1 receptor (IGF-1R)/Akt/β-catenin/cyclinD1 axis as well as tumor necrosis factor-α level and nuclear factor (NF)-kB activity. Furthermore, the treatment with CAPE replenished the antioxidant defense systems, superoxide dismutase (SOD) and reduced glutathione (GSH) with subsequent reduction in prostate tissue lipid peroxides. This study highlights the potential merit of CAPE-enriched propolis formulations to protect elderly men against the development of BPH. © 2018 IUBMB Life, 70(6):519-528, 2018.


Dissemination of Pseudomonas aeruginosa during lung infection in hamsters. Role of oxygen-induced lung injury.
Pseudomonas aeruginosa was inoculated into the lungs of normal and oxygen-exposed hamsters. Air-breathing animals developed focal bronchopneumonias but viable organisms were not recovered from the lungs after 3 days; bacteremia was not detected but P. aeruginosa was isolated from the livers of 3 of 12 animals with positive lung cultures. Pseudomonas aeruginosa infection shortened the survival time of oxygen-breathing hamsters, and organisms persisted in the lungs during oxygen exposure. Focal bronchopneumonias were uncommon in animals inoculated after 4 days of oxygen exposure; the predominant histopathologic finding was accentuation of diffuse alveolar damage and increased numbers of scattered neutrophils. Pseudomonas aeruginosa was recovered from the livers of 15 of 18 oxygen-exposed animals in which lung colony counts exceeded 10(3)/lung; only 8 of these animals had demonstrable bacteremia. The concentration of elongation factor-2 in the livers and lungs of oxygen-exposed animals was reduced as colony counts of P. aeruginosa increased in the lungs of infected animals, suggesting exotoxin A activity in these organisms. These findings suggest that bacterial superinfection of injured lungs may account for both worsening lung function and impaired function of other organs.


All-solid all-chalcogenide microstructured optical fiber.
The realization of an all-solid microstructured optical fiber based on chalcogenide glasses was achieved. The fiber presents As(2)S(3) inclusions selected as low refractive index material (n = 2.4) embedded in a As(38)Se(62) glass matrix (n = 2.8). The single mode regime of the fiber was demonstrated both theoretically by multipole method calculations and experimentally by near field measurements. Optical transmission measurements of the microstructured fiber and single index fibers made of the initial glasses reveal an excess of losses as high as 6-7 dB/m. This excess is not due to the guide geometry but can be explained by the presence of defects in the glass interface regions.


Palliative endoscopic retrograde biliary drainage for malignant biliary obstruction in Korea: A nationwide assessment.
Malignant biliary obstruction (MBO) is usually caused by biliary tract cancer or pancreatic cancer. This study was performed to summarize the current situation regarding palliative endoscopic retrograde biliary drainage (ERBD) for MBO in Korea and to determine its clinical significance by analyzing representative nationwide data. Patients that underwent palliative ERBD for MBO between 2012 and 2015 were identified using the Health Insurance Review and Assessment database, which covers the entire Korean population. We assessed clinical characteristics and complications and compared the clinical impacts of initial metal and plastic stenting in these patients. A total of 9,728 subjects (mean age, 65 ± 11.4 years; male, 61.4%) were identified and analyzed. The most common diagnosis was malignant neoplasm of liver and intrahepatic bile ducts (32.1%) and this was followed by extrahepatic or Ampulla of Vater cancer and pancreatic cancer. Initial plastic stent(s) placement was performed in 52.9% of the study subjects, and metal stent(s) placement was performed in 23.3%. The number of sessions of endoscopic retrograde cholangiopancreatography (ERCP) or percutaneous transhepatic biliary drainage (PTBD) was significantly higher in patients that underwent initial plastic stenting than in patients that underwent metal stenting (2.2 ± 1.7 vs 1.8 ± 1.4, P < 0.0001), but rates of post-ERCP pancreatitis, hospital days, and time to second ERCP or PTBD were not significantly different. This nationwide assessment study suggests that initial metal stenting is associated with fewer sessions of total ERCP or PTBD following the initial procedure, despite the preference for initial plastic stenting in Korea.


Functional involvement of RFVT3/SLC52A3 in intestinal riboflavin absorption.
Riboflavin, also known as vitamin B2, is transported across the biological membrane into various organs by transport systems. Riboflavin transporter RFVT3 is expressed in the small intestine and has been suggested to localize in the apical membranes of the intestinal epithelial cells. In this study, we investigated the functional involvement of RFVT3 in riboflavin absorption using intestinal epithelial T84 cells and mouse small intestine. T84 cells expressed RFVT3 and conserved unidirectional riboflavin transport corresponding to intestinal absorption. Apical [(3)H]riboflavin uptake was pH-dependent in T84 cells. This uptake was not affected by Na(+) depletion at apical pH 6.0, although it was significantly decreased at apical pH 7.4. The [(3)H]riboflavin uptake from the apical side of T84 cells was prominently inhibited by the RFVT3 selective inhibitor methylene blue and significantly decreased by transfection of RFVT3-small-interfering RNA. In the gastrointestinal tract, RFVT3 was expressed in the jejunum and ileum. Mouse jejunal and ileal permeabilities of [(3)H]riboflavin were measured by the in situ closed-loop method and were significantly reduced by methylene blue. These results strongly suggest that RFVT3 would functionally be involved in riboflavin absorption in the apical membranes of intestinal epithelial cells.


Stent implantation into the patent ductus arteriosus in cyanotic congenital heart disease with duct-dependent or diminished pulmonary circulation.
We assessed the efficacy and safety of stent implantation into the ductus arteriosus in infants with cyanotic heart disease. Ten patients with duct-dependent or diminished pulmonary circulation underwent cardiac catheterization with the aim of stent implantation. Coronary stents of 3.5-4 mm in diameter were used and successfully implanted in eight. All patients in whom stent implantation was successful had adequate relief of their cyanosis and were discharged home. During a mean follow-up of 9.3 months (median 10 months), one patient had evidence of restenosis of the stent at five months. Re-dilatation of the stenosed stent was attempted but was unsuccessful and a modified left Blalock-Taussig shunt was placed surgically. Stent implantation of ductus arteriosus in neonates and infants with a duct-dependent pulmonary circulation is an effective and safe palliative procedure. It may be an alternative to surgical systemic-pulmonary artery shunt in a select group of patients.


Retrospectively and prospectively modulated hippocampal place responses are differentially distributed along a common path in a continuous T-maze.
Hippocampal place responses can be prospectively or retrospectively modulated by the animal's future or prior trajectory. Two main hypotheses explain this. The "multiple-map hypothesis" switches between different maps for different trajectories (rate remapping). In contrast, in the "buffer hypothesis," the hippocampus encodes an ongoing representation that includes the recent past and/or the impending future choice. This study examines the distribution of prospective and retrospective responses distributed along a common path in a continuous T-maze (providing all four combinations of provenance and destination) during a visual discrimination task. The multiple-map hypothesis predicts either uniform distributions or concerted shifts about a task-decision relevant point, whereas the buffer hypothesis predicts a time-limited overexpression around choice points (with retrospective responses after the central arm entry point and prospective responses nearer its exit). Here bilateral recordings in the dorsal CA1 region of the rat hippocampus show that retrospective responses were twice as prevalent as prospective responses. Furthermore, retrospective and prospective modulations have distinct spatial distributions, with retrospective primarily in the first two-thirds of the central arm and prospective restricted to the last third. To test for possible trial-by-trial remapping in relation to the two-thirds transition point, data from the first and second halves of the sessions were compared. Backward drift of path-modulated activity was significant only for retrospective, but not prospective, fields. Thus, these data are more consistent with the buffer hypothesis. Retrospective and prospective modulation would then participate in a single hippocampal representation of spatial and behavioral context.


Third coronary artery.
The human hart is in most cases vascularized by two coronary arteries, the right and the left one. Supernumerary or added coronary arteries are also present sometimes. The aim of this paper is to ascertain presence of supernumerary coronary arteries that arise directly from aorta, their frequency, flow, way of branching, and possibly their anastomoses with other coronary arteries. Out of 25 hearts, examined by dissection, 8 of them (32%) had conal artery. According to our opinion, the most suitable term for this artery, in order to differ it clearly from the conal branch of the right coronary artery, is third coronary artery. One heart (4%) had four coronary arteries. Both supernumerary coronary arteries arose from the right aortic sinus. The third coronary artery represents a significant way of collateral coronary circulation, as it frequently anastomoses with anterior interventricular branch.


Electrolyzed water produced using carbon electrodes promotes the proliferation of normal cells while inhibiting cancer cells.
We previously developed electrolyzed water (EW) using carbon electrodes and estimated its ability to inhibit the proliferation of human cervical carcinoma HeLa cells. In this study, we found that EW-containing media could not only inhibit HeLa cell proliferation, but were also capable of promoting the proliferation of normal human dermal fibroblasts (NHDF). In addition, the developed EW could reduce cytochrome c, as demonstrated by the cytochrome c reduction assay. Interestingly, EW with a greater pH, which was unable to inhibit HeLa cell proliferation, completely lost the ability to reduce cytochrome c. Our results indicate that EW has opposite effects on cancer and normal cell proliferation and has the ability to reduce cytochrome c. Based on our findings, we suggest the possibility that the reducing capacity of our developed EW may be involved in the significant inhibition of HeLa cell proliferation.


Multiple forms of nuclear deoxyribonucleic acid polymerases and their relationship with the soluble enzyme.
1. DNA polymerase from nuclear and supernatant fractions of cultured mouse L929 cells was fractionated on columns of Sephadex G-200, Sepharose 4B and of DEAE-cellulose. Several peaks of activity are found on Sephadex chromatography and the distribution of activity between these depends on: (a) the source of the enzyme, i.e. nuclear or supernatant fraction; (b) the mode of extraction of the enzyme from the nucleus; (c) the amount of enzyme applied to the column. 2. The DNA polymerase activity in the lower-molecular-weight peaks (approximate molecular weights are 35000, 70000 and 140000) is firmly bound within the cell nucleus and shows a preference for native DNA as template, whereas the high-molecular-weight peak (peak I, molecular weight 250000 or greater) is found in supernatant fractions and shows greater activity with a denatured DNA template. 3. During periods of DNA synthesis the high-molecular-weight enzyme becomes more firmly bound within the nucleus. 4. Peak I enzymic activity is relatively unstable and is inhibited by thiol-blocking reagents and deoxycholate, but it is stimulated by univalent cations. 5. Very little endonuclease is present in the polymerase preparations, but a very active exonuclease and nucleoside diphosphokinase are present. On Sephadex chromatography, however, it was shown that the immediate precursors for DNA synthesis, at least by peak I enzyme, are the deoxyribonucleoside triphosphates. 6. Attempts to decrease the molecular weight of the peak I enzyme while still retaining activity failed.


Death Rate of Dental Anaesthesia.
Death was the most important side effect of anaesthesia in dentistry. In this article we reviewed more than 20 studies with adequate data focusing on death associated with dental procedures since 1955 and found 218 deaths out of 71,435,282 patients (3 deaths per 1,000,000 persons) with the mortality rate of 1:327,684. In addition, mortality rate per million has dropped to half (6.2 per 1,000,000 vs. 3 per 1,000,000) since 1955 till the last report in 2012 without any sex predilection. In children, most cases died in the age of two to five years. Hypoxia was the most common cause of death, and cardiovascular, respiratory, and endocrine disorders, hepatic cirrhosis, septicaemia, and bacterial endocarditis were the most frequent underlying systemic disease in deceased patients. Although rare death following general anaesthesia in dentistry, is a critical side effect mostly seen in patients with compromised health condition. Therefore, appropriate case selection in regard with patients' general health status as well as standard technical and equipment conditions are mandatory to diminish the risk of death during dental anaesthesia.


Analgesic Buxus alkaloids with Enhanced Selectivity for the Low-Voltage-Gated Calcium Channel Cav 3.2 over Cav 3.1 through a New Binding Mode.
Low-voltage-gated calcium channels (LVGCCs; Cav 3.1-3.3) represent promising drug targets for epilepsy, pain, and essential tremor. At present, modulators with heightened selectivity for a subtype of LVGCCs are still highly desired. In this study we explored three classes of Buxus alkaloids and identified 9(10/19)abeo-artanes Buxusemine H and Buxusemine L (BXSL) as an unprecedented type of Cav 3.2 inhibitors. Particularly, BXSL exhibited Cav 3.2 inhibition comparable to Z944, a non-subtype-selective LVGCCs inhibitor under clinical trial. While lacking specificity for Cav 3.3, BXSL showed a 30-fold selectivity of Cav 3.2 over Cav 3.1. As compared to several well-known inhibitors, the experimental and computational studies suggested BXSL exhibits a distinct binding mode to Cav 3.2, notably through the essential interaction with serine-1543 in domain III. Furthermore, BXSL showed minimal impact on various recombinant and native nociceptive ion channels, while significantly reducing the excitability of isolated mouse dorsal root ganglion neurons. Animal studies in wild-type and Cav 3.2 knock-out mice revealed that BXSL (5 mg/kg), by inhibiting Cav 3.2, exhibits an analgesic effect equivalent to Z944 (10 mg/kg) or mibefradil (10 mg/kg). Moreover, we proposed a structural rationale for the high selectivity of 9(10/19)abeo-artane-type alkaloids towards Cav 3.2 over Cav 3.1. This study introduces a novel analgesic agent and valuable molecular insight for structure-based innovative Cav 3.2 drug development.


[Factors influencing the use of mechanisms for coordinating healthcare levels in Colombia].
To analyse the factors influencing the use of mechanisms for the clinical coordination of two Colombian public healthcare networks' healthcare levels in Bogotá from the main social actors' perspective. This was a descriptive-interpretative, qualitative study of two public healthcare networks. Discussion groups and semi-structured interviews were used for collecting information. The approach involved two-stage theoretical sampling of a selection of centres operating at different healthcare levels and a selection of informants, including managers (n=19), healthcare employees (n=23) and administrative staff (n=20). Content analysis involved adopting a mixed method approach for generating categories, segmented by network, informant group and topic. Both networks had few mechanisms for enabling the clinical coordination of healthcare levels; information transfer mechanisms predominated and clinical management coordination mechanisms only dealt with maternal-perinatal care. Organisational factor-related complications were found regarding their use: lack of time, staff turnover, administrative use and technological deficiency. Employee/staff-related difficulties were due to lack of interest. These factors directly affected coordination with limited information transfer, patient follow-up and healthcare quality (diagnosis and treatment delays). The results highlighted the limited use of clinical coordination mechanisms in both public healthcare networks studied here, with problems in their use. Changes are required that affect directly organisational factors (time for coordination and working conditions) and professional factors (attitudes towards collaborative work).


The Stroop Color and Word Test.
The Stroop Color and Word Test (SCWT) is a neuropsychological test extensively used to assess the ability to inhibit cognitive interference that occurs when the processing of a specific stimulus feature impedes the simultaneous processing of a second stimulus attribute, well-known as the Stroop Effect. The aim of the present work is to verify the theoretical adequacy of the various scoring methods used to measure the Stroop effect. We present a systematic review of studies that have provided normative data for the SCWT. We referred to both electronic databases (i.e., PubMed, Scopus, Google Scholar) and citations. Our findings show that while several scoring methods have been reported in literature, none of the reviewed methods enables us to fully assess the Stroop effect. Furthermore, we discuss several normative scoring methods from the Italian panorama as reported in literature. We claim for an alternative scoring method which takes into consideration both speed and accuracy of the response. Finally, we underline the importance of assessing the performance in all Stroop Test conditions (word reading, color naming, named color-word).


Biobank participant support of newborn screening for disorders with variable treatment and intervention options.
We aimed to better understand biobank participant opinions of the benefits of newborn screening (NBS) for certain disorder types and how terminology used in NBS discourse might impact stakeholder opinion. We conducted a between-subjects randomized survey of 5840 members of the Northwestern University Biobank. The survey contained 12 scenarios, each describing a disorder and its treatment. For each scenario, we varied the terminology used to describe treatment options. One survey version used the term intervention and the other treatment. The outcome measured for each scenario was perceived benefit (for the infant) and importance of testing (for participants). Comparisons were made between participants and between scenarios. Ratings of benefit and importance were not influenced by the use of the term intervention versus treatment within scenarios. Nuances existed in ratings of benefit to the infant and importance to participants amongst scenarios. Participants were most likely to perceive benefit and importance in screening for a disorder if treatment/intervention offered a high chance of improved outcomes. While participants perceived benefit to the infant and importance to themselves in screening for most disorders, nuances in inter-scenario ratings suggest participants weighed availability and type of treatment/intervention in consideration of the benefits of NBS.


Cardiovascular profile of FPL 62129, a novel dihydropyridine calcium channel blocker, in anaesthetised dogs: a comparison with nifedipine.
The cardiovascular effects of FPL 62129, a novel dihydropyridine calcium channel blocker, were studied in chloralose-anaesthetised dogs in comparison with those of nifedipine. FPL 62129 and nifedipine produced dose-related falls in blood pressure and total peripheral resistance and were of similar potency in these respects. While the vasodilator effects of nifedipine were accompanied by reflex tachycardia, FPL 62129 reduced blood pressure without altering heart rate. Both compounds increased cardiac contractility and cardiac output. Nifedipine reduced PR interval duration of the ECG, whereas FPL 62129 produced no change. These differences between FPL 62129 and nifedipine are reflections of their respective cardiac profiles. In studies performed after cardiac autonomic block (to eliminate the contribution from reflexes), FPL 62129 showed marked activity at the sinus node and to a lesser extent at the atrioventricular node. Unlike nifedipine, FPL 62129 demonstrates vasodilator and direct decelerator properties over the same dose range, which accounts for its ability to lower blood pressure without causing reflex tachycardia in this model. The observed profile of FPL 62129 questions attempts to classify calcium channel blockers of the dihydropyridine type as selective vasodilators.


Peptides of the variable IgG domain as potential biomarker candidates in primary open-angle glaucoma (POAG).
Autoantibody profiling has gained increasing interest in the research field of glaucoma promising the detection of highly specific and sensitive marker candidates for future diagnostic purposes. Recent studies demonstrated that immune responses are characterized by the expression of congruent or similar complementarity determining regions (CDR) in different individuals and could be used as molecular targets in biomarker discovery. Main objective of this study was to characterize glaucoma-specific peptides from the variable region of sera-derived immunoglobulins using liquid chromatography--mass spectrometry (LC-MS)-based quantitative proteomics. IgG was purified from sera of 13 primary open-angle glaucoma patients (POAG) and 15 controls (CTRL) and subsequently digested into Fab and Fc by papain. Fab was further purified, tryptic digested and measured by LC-MS/MS. Discovery proteomics revealed in total 75 peptides of the variable IgG domain showing significant glaucoma-related level changes (P < 0.05; log2 fold change ≥ 0.5): 6 peptides were high abundant in POAG sera, whereas 69 peptides were low abundant in comparison to CTRL group. Via accurate inclusion mass screening strategy 28 IgG V domain peptides were further validated showing significantly decreased expression levels in POAG sera. Amongst others 5 CDR1, 2 CDR2 and 1 CDR3 sequences. In addition, we observed significant shifts in the variable heavy chain family distribution and disturbed κ/λ ratios in POAG patients in contrast to CTRL. These findings strongly indicate that glaucoma is accompanied by systemic effects on antibody production and B cell maturation possibly offering new prospects for future diagnostic or therapy purposes.


Accelerated pathological and clinical nephritis in systemic lupus erythematosus-prone New Zealand Mixed 2328 mice doubly deficient in TNF receptor 1 and TNF receptor 2 via a Th17-associated pathway.
TNF-alpha has both proinflammatory and immunoregulatory functions. Whereas a protective role for TNF administration in systemic lupus erythematosus (SLE)-prone (New Zealand Black x New Zealand White)F(1) mice has been established, it remains uncertain whether this effect segregates at the individual TNFR. We generated SLE-prone New Zealand Mixed 2328 mice genetically deficient in TNFR1, in TNFR2, or in both receptors. Doubly-deficient mice developed accelerated pathological and clinical nephritis with elevated levels of circulating IgG anti-dsDNA autoantibodies and increased numbers of CD4(+) T lymphocytes, especially activated memory (CD44(high)CD62L(low)) CD4(+) T cells. We show that these cells expressed a Th17 gene profile, were positive for IL-17 intracellular staining by FACS, and produced exogenous IL-17 in culture. In contrast, immunological, pathological, and clinical profiles of mice deficient in either TNFR alone did not differ from those in each other or from those in wild-type controls. Thus, total ablation of TNF-alpha-mediated signaling was highly deleterious to the host in the New Zealand Mixed 2328 SLE model. These observations may have profound ramifications for the use of TNF and TNFR antagonists in human SLE and related autoimmune disorders, as well as demonstrate, for the first time, the association of the Th17 pathway with an animal model of SLE.


Peritonitis in juvenile chronic arthritis. A report of 2 cases.
Peritonitis is a rare extra-articular manifestation of juvenile chronic arthritis (JCA) which, although mentioned in passing, seems not to have been documented previously. Two patients with systemic-onset JCA, 1 of whom had peritonitis in the early weeks of onset while the second developed peritonitis 10 years after onset, are described.


Protection by alpha-thalassaemia against Plasmodium falciparum malaria: modified surface antigen expression rather than impaired growth or cytoadherence.
We have attempted to determine the cellular mechanism by which alpha-thalassaemia may protect against Plasmodium falciparum malaria. Invasion and development of P. falciparum in the microcytic red cells of two-gene deletion forms of alpha-thalassaemia when measured morphologically or by [3H]hypoxanthine incorporation were normal compared to controls. Normal invasion rates were also observed following schizogony in thalassaemic red cells. Neither the addition of the oxidant menadione, 30% oxygen, nor modified medium, produced differential damage to parasites within thalassaemic cells. Furthermore, there were no significant differences in the binding of P. falciparum-parasitized alpha-thalassaemic and normal cells to C32 melanoma cells in vitro. However, when neoantigen expression on the surface of infected thalassaemic cells was estimated using a quantitative radiometric antiglobulin assay, clear differences were observed. It was found that alpha-thalassaemic cells bound higher levels of antibody from serum obtained from individuals living in a malaria endemic area than control normal red cells. The binding ratio for thalassaemic compared with controls was 1.69 on a cell-for-cell basis, and 1.97 when related to surface area. The binding of antibody from immune serum increased exponentially during parasite maturation. We also found increased binding of naturally occurring antibody present in non-immune serum to parasitized thalassaemic red cells which also increased during parasite maturation. We conclude that the protection afforded by thalassaemia against malaria may not reside in the ability of parasites to enter, grow or cytoadhere to endothelium in such cells, but may be related to immune recognition and subsequent clearance of parasitized red cells.


Fibrinogen and fibronectin as substrates for epidermal cell migration during wound closure.
Pieces of glass coverslip coated with human fibronectin or human fibrinogen were implanted under one margin of a skin wound on adult newt (Notophthalmus viridescens) hind limbs. In contrast to uncoated glass or glass coated with nest serum, bovine serum or bovine serum albumin, glass treated with either fibronectin or fibrinogen supported considerable epidermal cell migration. When optimal amounts of each protein were used, the amount of migration on fibrinogen-coated glass did not differ from the amount on fibronectin-coated glass or from the amount on the wound bed. Migration on a fibronectin substrate, could be blocked by treating the substrate with an antiserum against fibronectin just prior to implantation. Similarly, migration on a fibrinogen substrate could be blocked by exposing it to an antiserum against fibrinogen. While we have yet to determine it fibrinogen and fibronectin are interacting directly with the cell surface, our observations suggest that these two proteins may play an important role in wound closure by providing a suitable substrate for epithelial cell migration.


The effects of okra (Abelmoschus esculentus Linn.) on the cellular events associated with Alzheimer's disease in a stably expressed HFE neuroblastoma SH-SY5Y cell line.
It has been reported that persons carrying the H63D variant in their hemochromatosis (HFE) gene are at increased risk of Alzheimer's disease (AD). We investigated the possibility that okra (Abelmoschus esculentus) and quercetin could mitigate this risk factor by examining its effect on AD-associated cellular events in HFE stably expressing SH-SY5Y cells. Treatment of H63D HFE cells either with okra or quercetin significantly decreased reactive oxygen species (ROS), hydrogen peroxide (H2O2), and protein oxidation compared to untreated cells. The levels of tau phosphorylation at serine-199, serine-202, and serine-396 sites were also significantly decreased when cells were treated with okra. Exposure of the H63D and wild type (WT) cells to iron increased tau phosphorylation, but this response was decreased significantly when cells were treated with okra. The mechanism responsible for these changes appears to be related to decreased glycogen synthase kinase (GSK)-3β activity, an upstream signaling kinase of tau phosphorylation. We also established that okra treatment dramatically decreases intracellular iron levels in H63D cells compared to untreated cells. Our results provide important in vitro data on the effects of okra on various AD-associated cellular processes in H63D variant HFE cells. These results suggest okra may be beneficial in people expressing the H63D variant to reduce the risk of AD and other neurodegenerative diseases related to oxidative stress. Further in vivo studies would help confirm this.


Implementation of a Single Emulsion Mask for Three-Dimensional (3D) Microstructure Fabrication of Micromixers Using the Grayscale Photolithography Technique.
Three-dimensional (3D) microstructures have been exploited in various applications of microfluidic devices. Multilevel structures in micromixers are among the essential structures in microfluidic devices that exploit 3D microstructures for different tasks. The efficiency of the micromixing process is thus crucial, as it affects the overall performance of a microfluidic device. Microstructures are currently fabricated by less effective techniques due to a slow point-to-point and layer-by-layer pattern exposure by using sophisticated and expensive equipment. In this work, a grayscale photolithography technique is proposed with the capability of simultaneous control on lateral and vertical dimensions of microstructures in a single mask implementation. Negative photoresist SU8 is used for mould realisation with structural height ranging from 163.8 to 1108.7 µm at grayscale concentration between 60% to 98%, depending on the UV exposure time. This technique is exploited in passive micromixers fabrication with multilevel structures to study the mixing performance. Based on optical absorbance analysis, it is observed that 3D serpentine structure gives the best mixing performance among other types of micromixers.


Ni-Catalyzed Reductive Dicarbofunctionalization of Nonactivated Alkenes: Scope and Mechanistic Insights.
Olefins devoid of directing or activating groups have been dicarbofunctionalized here with two electrophilic carbon sources under reductive conditions. Simultaneous formation of one C(sp3)-C(sp3) and one C(sp3)-C(sp2) bond across a variety of unbiased π-systems proceeds with exquisite selectivity by the combination of a Ni catalyst with TDAE as sacrificial reductant. Control experiments and computational studies revealed the feasibility of a radical-based mechanism involving, formally, two interconnected Ni(I)/Ni(III) processes and demonstrated the different ability of Ni(I) species (Ni(I)I vs PhNi(I)) to reduce the C(sp3)-I bond. The role of the reductant was also investigated in depth, suggesting that a one-electron reduction of Ni(II) species to Ni(I) is thermodynamically favored. Further, the preferential activation of alkyl vs aryl halides by ArNi(I) complexes as well as the high affinity of ArNi(II) for secondary over tertiary C-centered radicals explains the lack of undesired homo- and direct coupling products (Ar-Ar, Ar-Alk) in these transformations.


Greater trochanteric pain syndrome: does imaging-identified pathology influence the outcome of interventions?
To assess the outcomes for patients seen in a rheumatology service presenting with features of the greater trochanteric pain syndrome (GTPS) and the impact of imaging results on the outcomes of treatment. Retrospective audit, using a phone interview was performed to establish links between results of imaging undertaken in the diagnostic work-up of patients with lateral hip pain and clinical outcomes for these patients. Patient perceptions of the effectiveness of interventions were also assessed. Forty-five patients were included (82% female, mean age 69.6 years). Sixty-nine percent underwent radiological work-up, including plain X-rays (55%), computed tomography scans (64%), magnetic resonance imaging (48%) and ultrasound (90%). Coexistent trochanteric bursitis (TB) and gluteal tendinopathy were the most commonly elucidated pathologies accounting for the symptomatic presentation of 40% of patients. Forty-one patients underwent some form of intervention, most commonly injection of local anesthetic and corticosteroid (LACS) into the region of the TB (87%), two-thirds of which were undertaken under radiological guidance. Pain reduction was maximal following the third injection, with a significantly better response to unguided interventions and levels of symptomatic relief following the first injection being a good indicator of the probability of complete remission. Radiological demonstration of isolated TB correlated with a greater reduction in lateral hip symptoms following LACS TB injections both in the immediate post-injection phase and in the long-term. The results of this audit suggest that the management of GTPS has reasonable patient outcomes; however, a prospective study with greater patient numbers is needed to confirm these results.


Molecular and Cellular Mechanisms for Trapping and Activating Emotional Memories.
Recent findings suggest that memory allocation to specific neurons (i.e., neuronal allocation) in the amygdala is not random, but rather the transcription factor cAMP-response element binding protein (CREB) modulates this process, perhaps by regulating the transcription of channels that control neuronal excitability. Here, optogenetic studies in the mouse lateral amygdala (LA) were used to demonstrate that CREB and neuronal excitability regulate which neurons encode an emotional memory. To test the role of CREB in memory allocation, we overexpressed CREB in the lateral amygdala to recruit the encoding of an auditory-fear conditioning (AFC) memory to a subset of neurons. Then, post-training activation of these neurons with Channelrhodopsin-2 was sufficient to trigger recall of the memory for AFC, suggesting that CREB regulates memory allocation. To test the role of neuronal excitability in memory allocation, we used a step function opsin (SFO) to transiently increase neuronal excitability in a subset of LA neurons during AFC. Post-training activation of these neurons with Volvox Channelrhodopsin-1 was able to trigger recall of that memory. Importantly, our studies show that activation of the SFO did not affect AFC by either increasing anxiety or by strengthening the unconditioned stimulus. Our findings strongly support the hypothesis that CREB regulates memory allocation by modulating neuronal excitability.


Iontophoresis for palmar and plantar hyperhidrosis.
Iontophoresis is a safe, efficacious, and cost-effective primary treatment of palmar and plantar hyperhidrosis. Decades of clinical experience and research show significant reduction in palmoplantar excessive sweating with minimal side effects. To get the best results from iontophoresis, health care professionals need to provide education on the mechanism of action and benefits, evidence of its use, and creation of a future patient-specific plan of care for continued treatments at home or in the physician's office. Iontophoresis may be combined with other hyperhidrosis treatments, such as topical antiperspirants and botulinum toxin injections.


Cannabis psychosis: examining the evidence for a distinctive psychopathology in a systematic and narrative review.
The term "cannabis psychosis" has become ubiquitous in the psychiatric literature. Few authors have described the precise psychopathology of this potentially distinct subtype of psychosis. Specifically, little attention has been paid to exploring whether cannabis psychosis is characterized by a psychopathology which is different from that of other types of psychosis. The purpose of this paper was to systematically review the literature for evidence of a specific constellation of symptoms which are consistently characteristic of cannabis psychosis within an inpatient psychiatric setting and to determine whether these combine to create a psychopathology which is distinct from that of other types of psychosis. Systematic review using Meta-analysis Of Observational Studies in Epidemiology (MOOSE) guidelines. 13 studies of the 439 identified met the inclusion criteria. Only eight studies had sufficient internal and external validity to allow comparison in a narrative format of the psychopathology present, compared with controls. Of these eight selected studies, seven reported at least one significant difference (p < .05) in the psychopathology of the cannabis group to the control group used as a comparator. This study should be interpreted with great caution and conclusions should not be generalized. These findings do not suggest that "cannabis psychosis" does not exist, only that from a psychopathological perspective it may not be qualitatively any different from other forms of psychosis. Future research in this area needs to focus on clarifying the definition or description of "cannabis psychosis" and the use of standardized robust experimental and/or observational designs to eliminate heterogeneity that may lead to inconclusive results.


[Comparative histology of mushroom bodies in carnivorous beetles of the suborder polyphaga (Insecta, Coleoptera)].
Mushroom bodies in beetles of the families Histeridae, Staphylinidae, Cantharidae, Trogossitidae, Peltidae, Cleridae, Malachiidae, and Coccinellidae are shown to be rather poorly developed. The calyx region of the mushroom bodies in these beetles never forms two separate cups, and the peduncular apparatus includes a unified shaft almost over its entire length. Only the pedunculus contains two separate shafts in a few cases. Two proliferative centers consisting of one to three neuroblasts are often found in each Kenyon cell group. The shift from carnivorous to feeding on pollen or leaves, which has taken place in some taxa, does not visibly affect the degree of mushroom body development.


Intellectual Property and the Politics of Public Good in COVID-19: Framing Law, Institutions, and Ideas during TRIPS Waiver Negotiations at the WTO.
To facilitate the manufacturing of COVID-19 medical products, in October 2020, India and South Africa proposed a waiver of certain WTO intellectual property (IP) provisions. After 18 months, a narrow agreement that did little for vaccine access passed the ministerial, despite the pandemic's impact on global trade, which the WTO is mandated to safeguard. The authors conducted a content analysis of WTO legal texts, key actor statements, media reporting, and the WTO's procedural framework to explore legal, institutional, and ideational explanations for the delay. IP waivers are neither legally complex nor unprecedented within WTO law, yet TRIPS waiver negotiations exceeded their mandated 90-day negotiation period by nearly 2 years. Waiver opponents and supporters engaged in escalating strategic framing, which justified and eventually secured political attention at head-of-state level, sidelining other pandemic solutions. The frames deployed discouraged consensus on a meaningful waiver, which ultimately favored the status quo that opponents preferred. WTO institutional design encouraged drawn-out negotiation while limiting legitimate players in debate to trade ministers, empowering narrow interest group politics. Despite global political attention, the WTO process contributed little to emergency vaccine production, suggesting a pressing need for reforms aimed at more efficient and equitable multilateral processes.


Accessibility to minors of cigarettes from vending machines--Broward County, Florida, 1996.
The sale of tobacco products to persons ages < 18 years has been prohibited by law in Florida since October 1992, and since May 1994, a statewide law in Florida has required retailers or owners of businesses that sell cigarettes or other tobacco products to post a conspicuous sign stating that tobacco sales to minors are illegal and that proof of age is required to purchase tobacco products. To assess the impact of these laws in Broward County (1990 population: 1,255,531) during February-March 1996, the Florida Atlantic University Department of Exercise Science/Wellness Education conducted studies of vendor compliance with laws enacted to prevent minors from gaining access to cigarettes through vending machines and to ensure that tobacco vendors comply with the sign statute. This report summarizes the findings of the assessment of access to cigarettes from vending machines, which indicated that approximately one third of such attempts by minors were successful.


Frame dislocation of body middle rings in endovascular stent tube grafts.
To understand the cause, and propose a mechanism for frame dislocation in endovascular grafts. Five tube grafts were explanted due to secondary distal leakage 15-21 months after operation. One bifurcated graft was removed during emergency operation after aortic rupture caused by secondary leakage. A second bifurcated graft was harvested from a patient with thrombotic occlusion of one limb, who died after transurethral prostatic resection. The inside of the grafts were examined endoscopically. The stent was inspected after removal of the fabric, broken ligatures were counted and examined by scanning electron microscopy. The fabric strength was tested by probe puncture. We found 17-44% of the stent ligatures of the body middle rings to be loose. The knots were intact. Degradation of the polyester textile was not observed. Continuous movements in the grafted aorta and blood pressure impose permanent stress to the stent frame and the polyester fabric resulting in morphological changes in the body middle ring of grafts. The clinical implications of the suture breakages are unknown although they may be related to distal secondary leakage in tube grafts.


Lighting the Way to Protein-Protein Interactions: Recommendations on Best Practices for Bimolecular Fluorescence Complementation Analyses.
Techniques to detect and verify interactions between proteins in vivo have become invaluable tools in functional genomic research. While many of the initially developed interaction assays (e.g., yeast two-hybrid system and split-ubiquitin assay) usually are conducted in heterologous systems, assays relying on bimolecular fluorescence complementation (BiFC; also referred to as split-YFP assays) are applicable to the analysis of protein-protein interactions in most native systems, including plant cells. Like all protein-protein interaction assays, BiFC can produce false positive and false negative results. The purpose of this commentary is to (1) highlight shortcomings of and potential pitfalls in BiFC assays, (2) provide guidelines for avoiding artifactual interactions, and (3) suggest suitable approaches to scrutinize potential interactions and validate them by independent methods.


Increased promyelocytic-derived microparticles: a novel potential factor for coagulopathy in acute promyelocytic leukemia.
The frequent serious bleeding and thrombotic complications in acute promyelocytic leukemia (APL) are major causes of early mortality, but the complex mechanisms causing the bleeding have not been completely elucidated. Because microparticles (MPs) are known to be elevated in thromboembolic disorders, we hypothesized a role for MPs in the pathogenesis of coagulopathy in APL. MPs were isolated from 30 APL patients and 20 healthy subjects and from cultured NB4/APL cells. The morphology of the MPs was examined, and they were quantified and analyzed for their thrombin-generating potential. We confirmed the existence of promyelocytic-derived MPs by morphology using transmission electron microscopy and laser scanning confocal microscopy. Counts of MPs in APL were elevated and were typically from promyelocytic cells (CD33(+) TF(+) MPs). Importantly, the CD33(+) MPs strongly correlated with patient leukocyte count (R = 0.64, p = 0.002) and D-dimer (R = 0.51, p = 0.0038). Moreover, the MPs from patients with APL decreased the coagulation times and induced thrombin generation. APL MP-associated thrombin generation was reduced by 54 % when the extrinsic pathway was blocked using an anti-human tissue factor (TF) antibody. However, neither anti-factor XI nor anti-tissue factor pathway inhibitor had any significant inhibitory effect. Our results show that the procoagulant state in APL is partially due to the TF-dependent procoagulant properties of circulating promyelocytic-derived MPs. TF(+) MPs may be a novel potential risk factor for coagulopathy in APL.


A longitudinal behavioral genetic analysis of the etiology of aggressive and nonaggressive antisocial behavior.
Developmental studies of antisocial behavior (ASB) have found two subgroups of behaviors, roughly described as aggressive and nonaggressive ASB. Theoretical accounts predict that aggressive ASB, which shows greater stability, should have high heritability. In contrast, nonaggressive ASB is very common in adolescence, shows less continuity, and should be influenced both by genes and shared environment. This study explored the genetic and environmental influences on aggressive and nonaggressive ASB in over 1,000 twin pairs aged 8-9 years and again at 13-14 years. Threshold models were fit to the data to incorporate the skew. In childhood, aggressive ASB was highly heritable and showed little influence of shared environment, whereas nonaggressive ASB was significantly influenced both by genes and shared environment. In adolescence, both variables were influenced both by genes and shared envirnmment. The continuity in aggressive antisocial behavior symptoms from childhood to adolescence was largely mediated by genetic influences, whereas continuity in nonaggressive antisocial behavior was mediated both by the shared environment and genetic influences. These data are in agreement with the hypothesis that aggressive ASB is a stable heritable trait as compared to nonaggressive behavior, which is more strongly influenced by the environment and shows less genetic stability over time.


Diversity, extinction risk and conservation of Malaysian fishes.
A total of 1951 species of freshwater and marine fishes belonging to 704 genera and 186 families are recorded in Malaysia. Almost half (48%) are currently threatened to some degree, while nearly one third (27%) mostly from the marine and coral habitats require urgent scientific studies to evaluate their status. Freshwater habitats encompass the highest percentage of threatened fish species (87%) followed by estuarine habitats (66%). Of the 32 species of highly threatened (HT) species, 16 are freshwater and 16 are largely marine-euryhaline species. Fish extinctions in Malaysia are confined to two freshwater species, but both freshwater and marine species are being increasingly threatened by largely habitat loss or modification (76%), overfishing (27%) and by-catch (23%). The most important threat to freshwater fishes is habitat modification and overfishing, while 35 species are threatened due to their endemism. Brackish-water, euryhaline and marine fishes are threatened mainly by overfishing, by-catch and habitat modification. Sedimentation (pollution) additionally threatens coral-reef fishes. The study provides recommendations to governments, fish managers, scientists and stakeholders to address the increasing and unabated extinction risks faced by the Malaysian fish fauna.


[Low frequency repetitive transcranial magnetic stimulation (rTMS) for the treatment of chronic tinnitus--are there long-term effects?].
Clinical, neurophysiological and neuroimaging data suggest that chronic tinnitus resembles neuropsychiatric syndromes characterised by focal brain activation. Low frequency repetitive transcranial magnetic stimulation (rTMS) has been proposed as an efficient method in treating brain hyperexcitability disorders. Patients suffering from chronic tinnitus underwent a [ (18)F]deoxyglucose-PET (positron emission tomography). Fusioning of the individual PET scan with the structural MRI-scan (magnetic resonance imaging, T1, MPRAGE) revealed an increased metabolic activation in the primary auditory cortex as target point for rTMS. A neuronavigational system enabled the positioning of the figure of eight coil in relation to the target area. rTMS (110 % motor threshold; 1 Hz; 2000 stimuli/day over 5 days) was performed using a placebo controlled cross-over design. Following active rTMS there was a moderate improvement of tinnitus perception. Treatment effects lasted up to six months in some patients. Neuronavigated rTMS offers new possibilities in the understanding and treatment of chronic tinnitus.


CD4+ T cell-mediated antigen-specific immunotherapy in a mouse model of cervical cancer.
A major agenda for tumor immunology is the generation of specific immune responses leading to the destruction of incipient and frank neoplasia. In this report, we show that a novel HPV16 E7 fusion protein can produce objective therapeutic responses against incipient cervical cancer in genetically engineered mice that express in the cervix the HPV16 early region genes implicated as causative agents in human cervical cancer. Although nonresponsive toward the HPV16 E7 oncoprotein in the CD8+ T-cell compartment by virtue of MHC haplotype, the mice were capable of mounting an induced CD4+ T-cell response against E7, and in addition developed spontaneous anti-E7 antibodies. HPV16/CD4-/- mice showed increased tumor burden indicative of CD4-mediated immune surveillance. Seeking to enhance the CD4 response, we immunized mice bearing incipient cervical cancer with a recombinant protein fusing E7 with a mycobacterial heat shock protein. The incidences of cervical carcinoma and of high-grade dysplasia (CIN 3) were consequently reduced by comparison to control mice. Thus, an HPV16 E7 immunogen holds promise for noninvasive treatment and prevention of human cervical cancer.


Assessing effects of disease genes and gene-environment interactions: the case-spouse design and the counterfactual-control analysis.
Assessing joint genetic and environmental contributions to disease risk is the central issue in many genetic epidemiological studies. To characterise the effects of a gene, the case-control study may suffer from the problem of population stratification bias. For a late onset disease, recruiting control subjects into case-parents and case-sibling studies may be difficult. Two novel approaches to analysing case-spouse data are introduced: the 1:1 case-counterfactual-control analysis (genotype swapping between the case and their spouse) and the 1:5 case-counterfactual-controls analysis (allele swapping). Both can be implemented using statistical packages that allow matched analysis (the conditional logistic regression) to yield valid estimates of the genotype relative risk, the gene-environment interaction parameter, the gene-sex interaction parameter, and the gene-environment-sex three factor interaction parameter (if desired), if certain assumptions are fulfilled. Because of the ease in recruiting subjects, and in collecting and analysing data, this approach makes a convenient tool for gene characterisation.


Somatic cell mating and segregation in chimeric frogs.
Both pentaploid and haploid cells were observed in a short-term culture of bone marrow of a diploid-triploid frog chimera. Apparently, diploid and triploid marrow cells fused to form "hybrid" pentaploid cells, which subsequently gave rise by somatic reduction to haploid daughter cells. The hybrid marrow cells and their haploid segregants are presumably at a selective disadvantage, as neither type of cell has been detected in the circulating blood of chimeric frogs.


Enzymatic degradation of human skin dermis revealed by fluorescence and reflectance spectroscopy.
Dermis is the major source of the fluorescence and light scattering of skin. Tumor-induced degradation of the dermis is expected to change the fluorescence and light scattering properties of skin. To investigate how these fluorescence and light scattering properties are changed, human skin dermis was degraded with enzymes to mimic tumor invasion. The enzymatic erosion process was investigated with fluorescence and reflectance spectroscopy. Dermis degradation by the enzymes resulted in a decrease in fluorescence emission and light scattering in the dermis. Fluorescence anisotropy, however, could not detect the change in the dermis induced by the enzyme treatments.


Discontinuing medications: a novel approach for revising the prescribing stage of the medication-use process.
Thousands of Americans are injured or die each year from adverse drug reactions, many of which are preventable. The burden of harm conveyed by the use of medications is a significant public health problem, and therefore, improving the medication-use process is a priority. Recent and ongoing efforts to improve the medication-use process have focused primarily on improving medication prescribing, and not much emphasis has been put on improving medication discontinuation. A formalized approach for rationally discontinuing medications is a necessary antecedent to improving medication safety and improving the nation's quality of care. This article proposes a conceptual framework for revising the prescribing stage of the medication-use process to include discontinuing medications. This framework has substantial practice and research implications, especially for the clinical care of older persons, who are particularly susceptible to the adverse effects of medications.


Epidemiology and clinical characteristics of the Measles outbreak in the Nylon Health District, Douala Cameroon: a retrospective descriptive cross sectional study.
Measles is a public health problem especially in South Asia and Africa. Nylon Health District has experienced two measles outbreaks over a period of three years. We hereby describe the epidemiology and clinical characteristics of the outbreak of February 2011. A retrospective descriptive cross sectional study was conducted in November 2011. All suspected measles cases according to the World Health Organization case definition line listed in the district service were included. Data was analyzed using Epi Info version 3.5.3 for Windows and Microsoft Office Excel 2010. An epidemic curve was drawn and proportions per variable category were estimated and presented in frequency tables. The outbreak started from the 4(th) to the 25(th) epidemiological week of 2011 with a peak on the 10(th) week after onset. The attack and case fatality rates were 34/100000 inhabitants and zero respectively. Females and infants aged 9-59 months represented 97(63.4%) and 75(49%) of cases respectively. Bonadiwoto health area alone had 81(52.9%) of cases. Of the 153 cases, only 34(22.2%) had a card-confirmed measles vaccination status. Active community surveillance permitted the identification of 42(27.5%) cases. Low measles vaccine coverage rate over the past years in the Nylon health district led to the accumulation of susceptible individuals which coupled with poor environmental conditions favoured inter-human spread of measles. Developing novel strategies to vaccinate every child, especially the Hard-to-Reach in the slums of the district will help to prevent future outbreaks.


Toxic encephalopathy caused by occupational exposure to 1, 2-Dichloroethane.
This study describes the clinical and neuroimaging features of five patients with 1, 2-Dichloroethane (DCE) toxic encephalopathy. From January 1st 1998 to June 30th 2009, five patients who were subsequently diagnosed with DCE toxic encephalopathy were admitted to our hospital. All were female workers who had been in contact with DCE and subsequently had had seizures or symptoms of intracranial hypertension, including headache, nausea, and vomiting. The cranial MRI showed extensive brain edema in either the subcortical white matter, bilateral globus pallidus, and cerebellar nucleus dendatus, or the cortices. Of the five patients in the study, three had vasogenic edema, one had cytotoxic edema, and one had both types of edema. Following treatment with steroids and mannitol for 3 to 10 weeks, all patients made either a partial or complete recovery. The imaging findings were resolved on a follow-up MRI. It is clear that occupational exposure to DCE can cause severe toxic encephalopathy. Moreover, extensive brain edema, secondary to blood-brain barrier damage or neuronal injury, is the major neuroimaging feature and the cause of clinical manifestations. Early diagnosis and prompt treatment leads to a good outcome.


Amorphization of Inorganic Solid Electrolyte Interphase Components and Its Impact on Enhancing Their Transport and Mechanical Properties.
The safety and cycle life of lithium-ion batteries (LIBs) are largely determined by the solid electrolyte interphase (SEI) formed on the surface of the anode. However, there is still a lack of understanding regarding the structure and properties of the individual SEI components. Among others, lithium oxide (Li2O), lithium carbonate (Li2CO3), and lithium fluoride (LiF) are known to be the main components of the inorganic SEI layer in conventional LIBs, but their intrinsic protective roles remain controversial. Herein, we present the transformational effects of their amorphous phase on the mechanical and transport characteristics, based on first-principles calculations. Our studies clearly demonstrate that their amorphous phases exhibit significantly improved Li-ion conductivity when compared to the crystalline structures. Additionally, among them, amorphous LiF emerges as a frontrunner for fast Li+ ion transportation, reversing the conventionally understood hierarchy. Under ambient conditions, the amorphous phases of LiF, Li2O, and Li2CO3 are thermodynamically unstable and tend to undergo recrystallization. However, this work highlights that exceptionally ductile and resilient amorphous phases can form if SEI formation and growth would involve some admixing of lithiophilic impurities like nitrogen (N) within the host matrices.


In situ formation and characterisation of singly ionised atomic europium in rare gas matrices--luminescence spectroscopy and MP2 calculations.
Irradiation of atomic europium isolated in the solid rare gases, with low intensity laser excitation of the y(8)P←a(8)S resonance transition at ca. 465 nm, is found to produce singly charged europium cations (Eu(+)) in large amounts in xenon and in smaller amounts in argon. Confirmation of the formation of matrix-isolated Eu(+) is obtained from characteristic absorption bands in the UV and in the visible spectral regions. The luminescence produced with excitation of the cation bands is presented in greatest detail for Eu/Xe and assigned. Excitation of the 4f(7)((8)S7/2)6p3/2 absorption bands of Eu(+) between 390 and 410 nm produces emission which is quite distinct from that resulting from excitation of the 4f(7)((8)S7/2)6p1/2 absorption (430 to 450 nm) features. The latter consists of narrow, resolved emission bands with Stokes shifts ten times smaller than the former. The observed spectral differences are discussed in relation to the different spatial symmetries of the p3/2 and p1/2 orbitals in these j-j coupled (7/2, 3/2)J and the (7/2, 1/2)J levels. Møller-Plesset calculations are conducted to obtain the molecular parameters of the neutral Eu-RG and cationic Eu(+)-RG diatomics (RG = Ar, Kr, Xe). From the short bond lengths and the strong binding energies obtained for the Eu(+)-RG species, these values suggest the isolation of the ion in small, possibly interstitial sites especially in xenon. In contrast, but consistent with previous work [O. Byrne and J. G. McCaffrey, J. Chem. Phys. 134, 124501 (2011)], the interaction potentials calculated herein for the Eu-RG diatomics suggest that the neutral Eu atom occupies tetra-vacancy (tv) and hexa-vacancy (hv) sites in the solid rare gas hosts. Possible reasons for the facile production of Eu(+) in the solid rare gases are discussed. The mechanism proposed is that atomic europium is also acting as an electron acceptor, providing a temporary trap for the ionised electron in the matrices.


Dietary habits and oral health in infancy and early childhood of Kenyan children at the Kenyatta National Hospital Paediatric Demonstration Unit.
The survey was conducted by the method of questionnaire in the Maternal Child Health (MCH) clinic. A total of ninety seven mothers brought their children to MCH clinic. The mean maternal age was 22 years, the youngest mother was 16 years old. The total number of children who attended the MCH clinic was 98 with an equal number of boys and girls. The age range was from 0.5-50 months with a mean age of 12 months. Most of the children were introduced to breastmilk substitutes or compliments from the age of 4 months but some of the children had been introduced to breastmilk compliments at an early age of one month. The commonly used breastmilk compliment at the age of one month was noted to be whole cow's milk, skimmed milk from Kenya Cooperative Creameries (KCC), cooked green bananas, ribena (only given to children of working mothers), fresh orange juice. The orange juice was noted to be given to children belonging to mothers who are either housewives or employed. For ages 2-3 months the commonly used breastmilk compliments or substitutes were noted to be maize meal, sorgum or millet porridge, whole cow's milk, KCC milk, ribena, orange juice, bananas and potatoes in order of preference. These same foods were given at ages 4 months up to 12 months. In addition pulses and eggs were introduced. The number of children who used the artificial formula was 33%. The method of feeding for the children attending the clinic was by using either bottle or spoon and cup. Twenty nine percent of these children were bottlefed.(ABSTRACT TRUNCATED AT 250 WORDS)


[Profile of silodosin].
Silodosin is a highly selective alpha1A-adrenoceptor antagonist approved for the treatment of the signs and symptoms of benign prostatic hyperplasia. Its clinical pharmacology profile offers a number of advantages, including uroselectivity, once-daily (QD) dosing, a standard dose of 8 mg QD that does not need to be adjusted according to age, and the feasibility of concomitant treatment with phosphodiesterase type 5 (PDE5) inhibitors and antihypertensive agents. Three phase 3 double-blind, randomised trials using the dosage regimen of 8 mg QD in > 800 patients have shown that silodosin is significantly more effective than placebo (p < 0.001) and at least as effective as tamsulosin (0.4 mg QD) in improving International Prostate Symptom Score (IPSS) total score, storage subscore, and voiding subscore. It is significantly more effective than tamsulosin in inducing simultaneous improvement of bothersome lower urinary tract symptoms such as incomplete emptying, frequency, and nocturia (p = 0.03). Safety data collected in 1581 patients exposed to chronic treatment with silodosin 8 mg QD have shown that the drug is safe and well tolerated. As was to be expected with a uroselective compound, cardiovascular effects have been minimal. The most common adverse reaction is "retrograde ejaculation" (anejaculation), which led to treatment discontinuation in only 3.9% of patients. The rare, drug class-related safety issue of intraocular floppy iris syndrome can be satisfactorily managed by warning patients simply to inform their ophthalmologist that they are or were on treatment with an alpha1-adrenoceptor blocker.


Ethnic differences in coronary artery calcium in a healthy cohort aged 60 to 69 years.
Measurement of coronary artery calcium (CAC) has been proposed as a screening tool, but CAC levels may differ according to race and gender. Racial/ethnic and gender distributions of CAC were examined in a randomly selected cohort of 60- to 69-year-old healthy subjects. Demographic, race/ethnicity (R/E), and clinical characteristics and assessment of CAC were collected. There were 723 white/European, 105 African-American, 73 Hispanic, and 67 East Asian subjects (597 men, 369 women) included in this analysis. Men had a significantly higher prevalence of any CAC (score>10) than women (76% vs 41%; p<0.0001). For men, the unadjusted odds of having any CAC was 2.2 (95% confidence interval [CI] 1.3 to 3.8) for whites compared with African-Americans. For women, CAC scores were not significantly different across ethnic groups. After adjustment for coronary risk factors, African-American and East Asian R/E remained associated with a lower prevalence of CAC in men (adjusted odds ratios [ORs] 0.33 and 0.47, respectively), as well as older age (OR 1.2, 95% CI 1.1 to 1.3), known hyperlipidemia (OR 1.7, 95% CI 1.1 to 2.7), and history of hypertension (OR 2.2, 95% CI 1.4 to 3.3). In women, Asian R/E (OR 2.5, 95% CI 1.1 to 5.7), history of smoking (adjusted OR 2.8, 95% CI 1.3 to 6.1), and known hyperlipidemia (adjusted OR 2.0, 95% CI 1.3 to 3.1) were associated with a higher prevalence of CAC independent of other risk factors. In conclusion, our data indicate that the presence of CAC varied significantly across selected race/ethnic groups independent of traditional cardiovascular risk factors.


The effect of breastfeeding on lung function at 12 and 18 years: a prospective cohort study.
The objective was to assess associations between duration of total and exclusive breastfeeding and lung function up to adolescence.A birth cohort (Melbourne Atopy Cohort Study) of 620 infants with a family history of allergic disease was recruited. Mothers were encouraged to breastfeed exclusively for 6 months. Lung function was assessed at 12 and 18 years of age. Associations between breastfeeding and lung function were investigated using multivariable linear regression and path analysis was used to assess the potential mediating factors.Duration of breastfeeding (total and exclusive) was not associated with most assessed lung function outcomes. However, there was a trend for increased pre-bronchodilator mid-expiratory flow (MEF) at both 12 (adjusted mean difference (95% CI) per week of breastfeeding of 10 (-1-20) mL·s(-1)) and 18 years (11 (-1-22) mL·s(-1)) (p-values of 0.07 and 0.08, respectively). There was a strong indirect effect of height on these observed associations.Duration of breastfeeding does not appear to greatly influence lung function outcomes in children with a family history of allergic diseases. Longer duration of exclusive breastfeeding may be associated with an increase in MEF, partly due to greater attained height of the child.


Tracking stem cells using magnetic nanoparticles.
Stem cell therapies offer great promise for many diseases, especially those without current effective treatments. It is believed that noninvasive imaging techniques, which offer the ability to track the status of cells after transplantation, will expedite progress in this field and help to achieve maximized therapeutic effect. Today's biomedical imaging technology allows for real-time, noninvasive monitoring of grafted stem cells including their biodistribution, migration, survival, and differentiation, with magnetic resonance imaging (MRI) of nanoparticle-labeled cells being one of the most commonly used techniques. Among the advantages of MR cell tracking are its high spatial resolution, no exposure to ionizing radiation, and clinical applicability. In order to track cells by MRI, the cells need to be labeled with magnetic nanoparticles, for which many types exist. There are several cellular labeling techniques available, including simple incubation, use of transfection agents, magnetoelectroporation, and magnetosonoporation. In this overview article, we will review the use of different magnetic nanoparticles and discuss how these particles can be used to track the distribution of transplanted cells in different organ systems. Caveats and limitations inherent to the tracking of nanoparticle-labeled stem cells are also discussed.


Rumination-focused cognitive behaviour therapy for non-responsive chronic depression: an uncontrolled group study.
One-third of patients with depression do not respond satisfactorily to treatment, and approximately 20% of all patients treated for depression develop a chronic depression. One approach to more effective treatment of chronic and treatment-resistant depression is to target rumination - an underlying mechanism implicated in the development and maintenance of depression. The purpose of this uncontrolled group study was to investigate the feasibility of individual rumination-focused cognitive behavioural therapy (RfCBT) for patients with chronic and treatment-resistant depression. A total of 10 patients with chronic and treatment-resistant depression were offered 12-16 individual sessions of RfCBT. The primary outcome was depressive symptoms as measured by Hamilton Depression Scale at pre-, post- and 3-month follow-up. Secondary symptoms measured included self-reported rumination and worry. There was a significant reduction in depressive symptoms (p < 0.05), rumination (p < 0.01) and worry (p < 0.5) from pre- to post-treatment. Half of the participants (n = 5) showed significant reliable change on levels of depressive symptoms post-treatment. The reduction in depressive symptoms, rumination and worry were maintained at follow-up. RfCBT was associated with significant reductions in depressive symptoms in a small sample with chronic and treatment-resistant depression. Despite limitations of being a small uncontrolled study with limited follow-up, these results are promising in a difficult to treat population. RfCBT warrants further systematic evaluation.


Extracellular vaccinia virus formation and cell-to-cell virus transmission are prevented by deletion of the gene encoding the 37,000-Dalton outer envelope protein.
There are two types of infectious vaccinia virus particles: intracellular naked virions and extracellular enveloped virions (EEV). To determine the biological role of the enveloped form of vaccinia virus, we produced and characterized a mutant that is defective in EEV formation. The strategy involved replacement by homologous recombination of the gene F13L, encoding a 37,000-Da protein (VP37) that is specific for the outer envelope of EEV, with a selectable antibiotic resistance marker, the Escherichia coli gpt gene. Initial experiments, however, suggested that such a mutation was lethal or prevented plaque formation. By employing a protocol consisting of high-multiplicity passages of intracellular virus from the transfected cells and then limiting dilution cloning, we succeeded in isolating the desired mutant, which was defective in production of plaques and extracellular virus but made normal amounts of intracellular naked virions. Electron microscopic examination indicated that the mutant virus particles, unlike wild type, were neither wrapped with Golgi-derived membranes nor associated with the cell surface. The absence of VP37 did not prevent the transport of the viral hemagglutinin to the plasma membrane but nevertheless abrogated both low-pH- and antibody-mediated cell fusion. These results indicate that VP37 is required for EEV formation and also plays a critical role in the local cell-to-cell transmission of vaccinia virus, perhaps via enveloped virions attached to or released from the cell membrane. By contrast, a mutated virus with a deletion of the K4L open reading frame, which is a homolog of the VP37 gene, was not defective in formation of plaques or EEV.


Ligation of the uterine artery and early postnatal food restriction - animal models for growth retardation.
Intrauterine growth restriction (IUGR) is one of the major causes of short stature in child- and adulthood. The cause of IUGR is unknown, however, an impaired uteroplacental function during the second half of human pregnancy might be an important factor, by affecting the programming of somatotropic axis and leading to postnatal growth failure into adulthood. Two rat models with perinatally induced growth retardation were used to examine the long-term effects of perinatal insults on growth. IUGR rats were prepared from pregnant dams, with a bilateral uterine artery ligation at day 17 of their pregnancy. Since the rat is relatively immature at birth, an early postnatal food restriction model was included as another model to broaden the time window of sensitive period of organogenesis. An individual growth curve was calculated of each animal (n = 813). From these individual growth curves the predicted growth curve for each experimental group was calculated by multilevel analysis. The proposed mathematical model allows us to estimate the growth potentials of these rat models with precision and could provide basic information to investigate the relationships among a number of other variables in future studies. Furthermore, we concluded that both pre- and early postnatal malnutrition leads to irreversible slowing down of postnatal growth.


Complications of glans penis augmentation.
Despite the controversy regarding penile augmentation (PA), glans penis augmentation (GPA) is used in penile reconstructive surgery in selected patients with small glans penis. Since 2003, injectable hyaluronic acid (HA) gel has been used for GPA. The attractiveness of HA gel fillers and interest in this technique have led to the use of other fillers for GPA, particularly irreversible fillers that improve the longevity of HA gels. Conversely, indirect glans augmentation via implantation of dermofat grafts or scaffolds between the corpus spongiosum and the distal tip of the corpus cavernosum is used to overcome poor longevity of the gel and glans surface undulation, which are the primary limitations of GPA using HA gel. Unfortunately, these implants form hard painful lumps over time in most patients and are difficult to remove. Blunt dissection is an invasive procedure that may cause vascular compromise and consequent glans necrosis in a few patients. We present our review regarding the current status of GPA for a more comprehensive understanding of this subject. Additionally, we have discussed a few complicated cases that were referred to us.


Aggregation-enhanced fluorescence in PEGylated phospholipid nanomicelles for in vivo imaging.
We report polymeric nanomicelles doped with organic fluorophores (StCN, (Z)-2,3-bis[4-(N-4-(diphenylamino)styryl)phenyl]-acrylonitrile), which have the property of aggregation-enhanced fluorescence. The fluorescent nanomicelles have two unique features: (1) They give much brighter fluorescence emission than mono-fluorophores. (2) The nanomicelles with amphiphilic copolymers [e.g., phospholipids-PEG (polyethylene glycol)] make the encapsulated fluorophores more stable in various bio-environments and easy for further conjugation with bio-molecules. After chemical and optical characterization, these fluorescent nanomicelles are utilized as efficient optical probes for in vivo sentinel lymph node (SLN) mapping of mice. The StCN-encapsulated nanomicelles, as well as their bioconjugates with arginine-glycine-aspartic acid (RGD) peptides, are used to target subcutaneously xenografted tumors in mice, and in vivo fluorescence images demonstrate the potential to use PEGylated phospholipid nanomicelles with aggregation-enhanced fluorescence as bright nanoprobes for in vivo diagnosis of tumors.


Association of HMOX1 gene promoter polymorphisms with hyperbilirubinemia in the early neonatal period.
Heme oxygenase (HO) is the rate-limiting enzyme in the heme degradation pathway that produces bilirubin. The promoter region of human heme oxygenase-1 (HMOX1) contains a polymorphic (GT)n repeat that can regulate gene expression. Here, we investigated the association of (GT)n repeat length in the HMOX1 promoter region with neonatal hyperbilirubinemia in a population of Japanese term neonates. Using polymerase chain reaction and fragment analysis, we determined the number of (GT)n repeats in 149 Japanese neonates. To omit the effects of the G71R mutation in uridine diphosphoglucuronosyltransferase on hyperbilirubinemia, we excluded 41 neonates with the G71R mutation. As a result, 25 neonates with hyperbilirubinemia and 83 non-hyperbilirubinemic controls were included in this prospective case-control study. Allele and genotype frequencies of (GT)n repeats in the HMOX1 gene were compared between hyperbilirubinemic and non-hyperbilirubinemic control neonates. The prevalence of short alleles (< 22 (GT)n repeats) was significantly higher in hyperbilirubinemic than in control neonates (18% vs 7%, P = 0.015). Hyperbilirubinemia was more frequent in homozygous or heterozygous short allele carriers than control neonates (28% vs 11%, respectively, P = 0.03). Possession of short alleles was significantly associated with the development of neonatal hyperbilirubinemia (OR, 3.1; 95%CI: 1.03-9.53). Infants carrying short alleles (< 22 (GT)n repeats) in the HMOX1 gene promoter region appear to be at a higher risk for developing neonatal hyperbilirubinemia.


Fluorescence resonance energy transfer (FRET)-based subcellular visualization of pathogen-induced host receptor signaling.
Bacteria-triggered signaling events in infected host cells are key elements in shaping the host response to pathogens. Within the eukaryotic cell, signaling complexes are spatially organized. However, the investigation of protein-protein interactions triggered by bacterial infection in the cellular context is technically challenging. Here, we provide a methodological approach to exploit fluorescence resonance energy transfer (FRET) to visualize pathogen-initiated signaling events in human cells. Live-cell microscopy revealed the transient recruitment of the Src family tyrosine kinase Hck upon bacterial engagement of the receptor carcinoembryonic antigen-related cell adhesion molecule 3 (CEACAM3). In cells expressing a CEACAM3 variant lacking the cytoplasmic domain, the Src homology 2 (SH2) domain of Hck (Hck-SH2) was not recruited, even though bacteria still bound to the receptor. FRET measurements on the basis of whole cell lysates revealed intimate binding between Hck-SH2 (using enhanced yellow fluorescent protein (YPet)-Hck-SH2) and the tyrosine-phosphorylated enhanced cyan fluorescent protein-labeled cytoplasmic domain of wild-type CEACAM3 (CEACAM3 WT-CyPet) and a flow cytometry-based FRET approach verified this association in intact cells. Using confocal microscopy and acceptor photobleaching, FRET between Hck-SH2 and CEACAM3 was localized to the sites of bacteria-host cell contact. These data demonstrate not only the intimate binding of the SH2 domain of Hck to the tyrosine-phosphorylated cytoplasmic domain of CEACAM3 in intact cells, but furthermore, FRET measurements allow the subcellular localization of this process during bacterial infection. FRET-based assays are valuable tools to resolve bacteria-induced protein-protein interactions in the context of the intact host cell.


Long-term health impacts of COVID-19 among 242,712 adults in England.
The COVID-19 pandemic is having a lasting impact on health and well-being. We compare current self-reported health, quality of life and symptom profiles for people with ongoing symptoms following COVID-19 to those who have never tested positive for SARS-CoV-2 infection and those who have recovered from COVID-19. Overall, 276,840/800,000 (34·6%) of invited participants took part. Mental health and health-related quality of life were worse among participants with ongoing persistent symptoms post-COVID compared with those who had never had COVID-19 or had recovered. In this study, median duration of COVID-related symptoms (N = 130,251) was 1·3 weeks (inter-quartile range 6 days to 2 weeks), with 7·5% and 5·2% reporting ongoing symptoms ≥12 weeks and ≥52 weeks respectively. Female sex, ≥1 comorbidity and being infected when Wild-type variant was dominant were associated with higher probability of symptoms lasting ≥12 weeks and longer recovery time in those with persistent symptoms. Although COVID-19 is usually of short duration, some adults experience persistent and burdensome illness.


Biochemistry and molecular biology techniques for person characterization.
Using the traditional serological tests and the most novel techniques for DNA fingerprinting, forensic scientists scan different traits that vary from person to person and use the data to include or exclude suspects based on matching with the evidence obtained in a criminal case. Although the forensic application of these methods is well known, the procedures and techniques used to obtain these results are not so well studied. Here, we report a laboratory exercise aimed to familiarize the students in several of the methods and markers employed for people individualization. With this exercise, students have to characterize themselves and an unknown sample according to the following characteristics: ABO blood group, presence or absence of Y chromosome, and their polymorphism for a variable number of tandem repeats (VNTR). This exercise has been originally developed for students of the course "Techniques in Criminology" within the Bachelor of Criminology. Although the schedule and procedures have been addressed to students who are faced for their first time with a laboratory of molecular biology, the exercise can also be adapted to students in more specialized courses and studies.


Molecular Diagnosis of Hemorrhagic Fever with Renal Syndrome Caused by Puumala Virus.
Rodent-borne hantaviruses cause two severe acute diseases: hemorrhagic fever with renal syndrome (HFRS) in Eurasia, and hantavirus pulmonary syndrome (HPS; also called hantavirus cardiopulmonary syndrome [HCPS]) in the Americas. Puumala virus (PUUV) is the most common causative agent of HFRS in Europe. Current routine diagnostic methods are based on serological analyses and can yield inconclusive results. Hantavirus-infected patients are viremic during the early phase of disease; therefore, detection of viral RNA genomes can be a valuable complement to existing serological methods. However, the high genomic sequence diversity of PUUV has hampered the development of molecular diagnostics, and currently no real-time reverse transcription-quantitative (RT)-PCR assay is available for routine diagnosis of HFRS. Here, we present a novel PUUV RT-PCR assay. The assay was validated for routine diagnosis of HFRS on samples collected in Sweden during the winter season from 2013 to 2014. The assay allowed detection of PUUV RNA in 98.7% of confirmed clinical HFRS samples collected within 8 days after symptomatic onset. In summary, this study shows that real-time RT-PCR can be a reliable alternative to serological tests during the early phase of HFRS.


3D microstructural architecture of muscle attachments in extant and fossil vertebrates revealed by synchrotron microtomography.
Firm attachments binding muscles to skeleton are crucial mechanical components of the vertebrate body. These attachments (entheses) are complex three-dimensional structures, containing distinctive arrangements of cells and fibre systems embedded in the bone, which can be modified during ontogeny. Until recently it has only been possible to obtain 2D surface and thin section images of entheses, leaving their 3D histology largely unstudied except by extrapolation from 2D data. Entheses are frequently preserved in fossil bones, but sectioning is inappropriate for rare or unique fossil material. Here we present the first non-destructive 3D investigation, by propagation phase contrast synchrotron microtomography (PPC-SRµCT), of enthesis histology in extant and fossil vertebrates. We are able to identify entheses in the humerus of the salamander Desmognathus from the organization of bone-cell lacunae and extrinsic fibres. Statistical analysis of the lacunae differentiates types of attachments, and the orientation of the fibres, reflect the approximate alignment of the muscle. Similar histological structures, including ontogenetically related pattern changes, are perfectly preserved in two 380 million year old fossil vertebrates, the placoderm Compagopiscis croucheri and the sarcopterygian fish Eusthenopteron foordi. We are able to determine the position of entheses in fossil vertebrates, the approximate orientation of the attached muscles, and aspects of their ontogenetic histories, from PPC-SRµCT data. Sub-micron microtomography thus provides a powerful tool for studying the structure, development, evolution and palaeobiology of muscle attachments.


Burrow webs: Clawing the surface of interactions with burrows excavated by American badgers.
Ecosystem engineers are organisms that influence their environment, which includes alterations leading to habitat provisioning for other species. Perhaps the most well-examined guild of species provisioning habitat for other species is tree cavity excavators or woodpeckers (Picidae). Many studies have examined the suite of secondary cavity users that rely on woodpeckers, and how the ecological network of secondary users, collectively referred to as the nest web, changes across communities. Despite similar habitat provisioning processes, fewer studies have assessed the suite of species associated with burrowers providing access to subterranean habitat. Here, we begin to characterize the burrow web provisioned by American badgers (Taxidea taxus) and evaluate the diversity and frequency of species interactions we detected at abandoned badger burrows in Wyoming, USA. We deployed camera traps at 23 badger burrows and identified interactions with the burrow by birds, mammals, and reptiles. Overall, we discovered 31 other species utilizing badger burrows, consisting of 12 mammals, 18 birds, and 1 reptile. Mammals, other than American badgers themselves and other fossorial species such as ground squirrels (Urocitellus sp.), frequently using burrows included mice (Peromyscus sp.), long-tailed weasel (Mustela frenata), pygmy rabbit (Brachylagus idahoensis), and desert cottontail (Sylvilagus audubonii). Of the 18 bird species detected, most accounted for <5% of overall detections, besides chipping sparrows (Spizella passerina) at 7.2%-11.5% of detections. The most common category of detection by bird species was foraging, contrary to mammals, which used the burrow frequently and were commonly observed entering and exiting the burrow. This work provides additional context on the ecological role of American badgers within their environment. More broadly, this work scratches the surface of many remaining questions to explore with the aim of advancing our understandings about burrow webs across the diversity of burrowing species and the communities in which they occur.