Successes and gaps in the HIV cascade of care of a high HIV prevalence setting in Zimbabwe: a population-based survey.
Gutu, a rural district in Zimbabwe, has been implementing comprehensive HIV care with the support of Médecins Sans Frontières (MSF) since 2011, decentralizing testing and treatment services to all rural healthcare facilities. We evaluated HIV prevalence, incidence and the cascade of care, in Gutu District five years after MSF began its activities. A cross-sectional study was implemented between September and December 2016. Using multistage cluster sampling, individuals aged ≥15 years living in the selected households were eligible. Individuals who agreed to participate were interviewed and tested for HIV at home. All participants who tested HIV-positive had their HIV-RNA viral load (VL) measured, regardless of their antiretroviral therapy (ART) status, and those not on ART with HIV-RNA VL ≥ 1000 copies/mL had Limiting-Antigen-Avidity EIA Assay for cross-sectional estimation of population-level HIV incidence. Among 5439 eligible adults ≥15 years old, 89.0% of adults were included in the study and accepted an HIV test. The overall prevalence was 13.6% (95%: Confidence Interval (CI): 12.6 to 14.5). Overall HIV-positive status awareness was 87.4% (95% CI: 84.7 to 89.8), linkage to care 85.5% (95% CI: 82.5 to 88.0) and participants in care 83.8% (95% CI: 80.7 to 86.4). ART coverage among HIV-positive participants was 83.0% (95% CI: 80.0 to 85.7). Overall, 71.6% (95% CI 68.0 to 75.0) of HIV-infected participants had a HIV-RNA VL < 1000 copies/mL. Women achieved higher outcomes than men in the five stages of the cascade of care. Viral Load Suppression (VLS) among participants on ART was 83.2% (95% CI: 79.7 to 86.2) and was not statistically different between women and men (p = 0.98). The overall HIV incidence was estimated at 0.35% (95% CI 0.00 to 0.70) equivalent to 35 new cases/10,000 person-years. Our study provides population-level evidence that achievement of HIV cascade of care coverage overall and among women is feasible in a context with broad access to services and implementation of a decentralized model of care. However, the VLS was relatively low even among participants on ART. Quality care remains the most critical gap in the cascade of care to further reduce mortality and HIV transmission.


Expression of the plasma prekallikrein gene: utilization of multiple transcription start sites and alternative promoter regions.
The plasma prekallikrein gene is expressed in many different human tissues at distinctly different levels and therefore tissue-specific control of the gene transcription is likely. In this study we demonstrate that transcription of the plasma prekallikrein gene can be initiated at multiple sites, for which at least four different promoters are utilized. A comparison of the genomic and mRNA sequences of mouse plasma prekallikrein revealed that the sequence segment that was formerly regarded as the first exon of the mouse plasma prekallikrein gene consists of three exons, with the first exon localized 14.2 kbp upstream of the translation start. For the rat and human plasma prekallikrein genes, in silico analysis suggested an analogous exon-intron organization. Determination of the transcription start sites showed that in both mouse and human, the proximal and distal regions could be utilized for transcription initiation; however, the proximal region is preferred. A deletion mutation analysis of the proximal promoter region using a 1.7-kbp segment revealed a strong activating region immediately upstream of the known mRNA, followed by both a modest repressor and an enhancer region.


A universal approach for calculating the Judd-Ofelt parameters of RE3+ in powdered phosphors and its application for the β-NaYF4:Er3+/Yb3+ phosphor derived from auto-combustion-assisted fluoridation.
It is difficult to calculate the Judd-Ofelt (J-O) parameters for trivalent rare earth (RE)-doped powders due to the unavailable absorption spectrum that is necessarily used in the conventional J-O calculation procedure. In this study, a universal method starting from the diffuse-reflection spectrum for calculating the J-O parameters of RE3+-doped powdered samples was proposed. In this proposed method, by taking the Kubelka-Munk function into account, the absorption cross-section spectrum was derived from the diffuse-reflection spectrum in the RE3+-doped powdered sample using the connection between the absorption cross section and the radiative transition rate of RE3+. Then, the J-O parameters might be calculated from the absorption cross-section spectrum via the traditional J-O calculation technique. The NaYF4:Er3+/Yb3+ and NaYF4:Er3+ phosphors were prepared via an auto-combustion-assisted fluoridation technique, and the J-O calculation was carried out for the obtained samples. The obtained J-O parameters were compared with those reported in the literature and also verified by comparing the calculated radiative transition lifetimes with the experimental values. Finally, it was deduced that the proposed J-O calculation route was practicable.


The Design and Rationale of the Trail1 Trial: A Randomized Double-Blind Phase 2 Clinical Trial of Pirfenidone in Rheumatoid Arthritis-Associated Interstitial Lung Disease.
Rheumatoid arthritis (RA) is the most common of the connective tissue diseases (CTD), affecting up to 0.75% of the United States (U.S.) population with an increasing prevalence. Interstitial lung disease is prevalent and morbid condition in RA (RA-ILD), affecting up to 60% of patients with RA, leading to premature death in 10% and accruing an average of US$170,000 in healthcare costs per patient over a 5-year period. Although there have been significant advances in the management of this joint disease, there are no ongoing randomized clinical trials looking at pharmacologic treatments for RA-ILD, and there currently are no U.S. Food and Drug Administration-approved drugs for RA-ILD. We describe the Treatment for Rheumatoid Arthritis and Interstitial Lung Disease 1 (TRAIL1) trial, a multicenter randomized, double-blind, placebo-controlled, phase 2 study of the safety, tolerability and efficacy of pirfenidone in patients with RA-ILD. The study will enroll approximately 270 subjects across a network of sites who have RA and ILD as defined by a fibrotic abnormality involving greater than 10% of the lung parenchyma. The primary endpoint of the study is the incidence of the composite endpoint of decline in percent predicted forced vital capacity of 10 or greater or death during the 52-week study period. A number of secondary and exploratory endpoints have been chosen to evaluate the safety and efficacy in different domains. The TRAIL1 trial is designed to evaluate the safety and efficacy of pirfenidone in RA-ILD, a disease with significant impact on patients' quality of life and outcome. In addition to investigating the safety and efficacy of pirfenidone, this trial looks at a number of exploratory endpoints in an effort to better understand the impact of therapy on areas such as changes in quantitative high-resolution computed tomography scores and a patient's quality of life. Biospecimens will be collected in order to investigate biomarkers that could potentially predict the subtype of disease, its behavior over time, and its response to therapy. Finally, by creating a network of institutions and clinician investigators with an interest in RA-ILD, this trial will pave the way for future studies of investigational agents in an effort to reduce or eliminate the burden of disease for those suffering from RA-ILD. Genentech, a member of the Roche Group. Clinicaltrials.gov, identifier NCT02808871.


Premature ventricular complexes as a first manifestation in left ventricular non-compaction cardiomyopathy: A case report and focused review of the literature.
Left ventricular non-compaction (LVNC) cardiomyopathy is an uncommon unclassified or genetic myocardial disorder. Frequent premature ventricular complexes (PVCs) as unique finding in LVNC cardiomyopathy are rare. We report a case of a 36-year-old woman in whom isolated LVNC was diagnosed due to an incidental finding of PVCs in pre-operative consultation.


Safety of TCMCB07, a melanocortin-4 antagonist peptide, in dogs with naturally occurring cachexia.
The melanocortin 4 antagonist TCMCB07 is safe and effective in reversing cachexia caused by sepsis or cancer in rodents. The safety and pharmacokinetics of TCMCB07 are demonstrated in healthy beagle dogs. The objectives of this study were to investigate the safety, peak plasma concentrations, and potential for efficacy of TCMCB07 in pet dogs with naturally occurring cachexia over a 4-week time period. Fourteen dogs with cachexia of any underlying cause, except cancer of the oral cavity or gastrointestinal tract, were eligible for enrollment with informed client consent. This study was a prospective, 1-armed open-label trial. Physical examination, complete blood count, chemistry panel, and owner-assessed quality of life surveys were checked at weeks 1, 2, and 4. Due to potential for bradycardia and hypotension, Holter monitoring and blood pressure evaluations were scheduled at pre-enrollment and week 4. Fourteen dogs completed the trial. Significant changes detected included increased mean body weight (18.6-19.5 kg, P < .02), increased body condition score (median Tufts 5-point thin dog scale score P < .004 and WSAVA muscle condition score P < .02) and increased mean blood urea nitrogen (21.79-30.43 mg dL-1 , P < .004). On quality of life surveys, pet owners perceived their dog appeared to be panting less (P < .002) and that the general health improved (P < .03). Four dogs had a change in coat pigmentation. The peak plasma concentration of TCMCB07 in cachectic dogs was similar to that in healthy beagle dogs. TCMCB07 was safe and has potential efficacy in pet dogs with cachexia.


Ultraviolet lighting during orthopaedic surgery and the rate of infection.
Ultraviolet lighting is an alternative to laminar airflow in the operating room that may be as effective for lowering the number of environmental bacteria and possibly lowering infection rates by killing the bacteria rather than simply reducing the number at the operative site. The purpose of the present study was to compare the infection rates following joint replacement procedures performed by one orthopaedic surgeon with and without the use of ultraviolet lighting. From July 1986 to July 2005, one surgeon performed 5980 total joint replacements at one facility. In September 1991, ultraviolet lighting was installed in the operating rooms. All procedures that were performed before the installation of the ultraviolet lighting utilized horizontal laminar airflow, whereas all procedures that were performed after that date utilized ultraviolet lighting without laminar airflow. Factors associated with the rate of infection were analyzed. Over a nineteen-year period, forty-seven infections occurred following 5980 joint replacements. The infection rate without ultraviolet lighting (and with laminar airflow) was 1.77%, and the infection rate with ultraviolet lighting was 0.57% (p < 0.0001). The odds of infection were 3.1 times greater for procedures performed without ultraviolet lighting (and with laminar airflow) as compared with those performed with only ultraviolet lighting (p < 0.0001). The infection rate associated with total hip replacement decreased from 1.03% to 0.72% (p = 0.5407), and the infection rate associated with total knee replacement decreased from 2.20% to 0.50% (p < 0.0001). Revision surgery, previous infection, age, total body mass index, use of cement, disease, and diagnosis were not associated with an elevated infection rate. When appropriate safety precautions are taken, ultraviolet lighting appears to be an effective way to lower the risk of infection in the operating room during total joint replacement surgery.


Overview of systematic reviews on invasive treatment of stable coronary artery disease.
The aim of the study was to evaluate the validity of the systematic reviews as a source of best evidence and to present and interpret the evidence of the systematic reviews on effectiveness of surgery and percutaneous interventions for stable coronary artery disease. Electronic databases were searched without language restriction from January 1966 to March 2004. The databases used included the Cochrane Database of Systematic Reviews, the Cochrane Central Register of Controlled Trials, DARE, the Health Technology Assessment Database, MEDLINE(R), MEDLINE(R) In-Process & Other Non-Indexed Citations. We included systematic reviews of randomized clinical trials on patients with stable coronary heart disease undergoing percutaneous coronary intervention or coronary artery bypass surgery in comparison with medical treatment or a comparison between invasive techniques. At least one of the following outcomes had to be reported: death, myocardial infarction, angina pectoris, revascularization. The methodological quality was assessed using a modified version of the scale devised by Oxman and Guyatt (1991). A standardized data-extraction form was used. The method used to evaluate clinical relevance was carried out with updated method guidelines from the Cochrane Back Research Group. Quantitative synthesis of the effectiveness data is presented. We found nineteen systematic reviews. The median score of validity was 13 points (range, 6-17 points), with a maximum of 18 points. Coronary artery bypass surgery gives better relief of angina, and the need for repeated procedures is reduced after bypass surgery compared with percutaneous interventions. There is inconsistent evidence as to whether bypass surgery improves survival compared with percutaneous intervention. A smaller need for repeated procedures exists after bare metal stent and even more so after drug-eluting stent placement than after percutaneous intervention without stent placement. However, according to the current evidence, these treatment alternatives do not differ in terms of mortality or myocardial infarction. We found some high-quality systematic reviews. There was evidence on the potential of invasive treatments to provide symptomatic relief. Surgery seems to provide a longer-lasting effect than percutaneous interventions with bare metal stents or without stents. Evidence in favor of drug-eluting stents so far is based on short-term follow-up and mostly on patients with single-vessel disease.


Significance of Perfluoroalkyl Substances (PFAS) in Food Packaging.
Food safety authorities and the food industry are focused on uses of perfluoroalkyl substances (PFAS) in various food-contact packaging applications. Not widely known until recently, certain PFAS occur in paper-based packaging materials typically at parts-per-billion to parts-per-million concentrations. These substances are nonintentionally added substances (NIAS) and are attributed to residues from recycled fiber and paperboard used in the manufacture of new food packaging products. Low concentration PFAS detection has generated debate in the food industry and among scientific and governmental organizations about understanding their significance in food-contact products because certain PFAS are intentionally added to some food packaging materials. Distinguishing between both sources of PFAS in food packaging is essential for regulatory compliance purposes. In this paper, we describe ongoing research using contact angle measurement analysis to determine limits of performance (LOP) for perfluorocarboxylic acids (PFCAs) (C4, C6, C8, and C10) on the surface of recycled paper packaging materials. We find that the LOP concentrations for PFCAs ranged from 37 ppm (C10) to higher than 1238 ppm (C4). Because there is no economic justification for the presence of PFAS that do not provide functional performance, these LOP concentrations can reliably be considered as NIAS thresholds. This analytical method and the resulting test data are able to differentiate the source of PFAS in food packaging. Future research will broaden the test method to include measurements of fluorotelomer, sulfonamide, and fluoropolymer substances to develop a more comprehensive understanding of PFAS performance and NIAS concentration thresholds. Integr Environ Assess Manag 2021;17:7-12. © 2020 SETAC.


Sertindole, a potent antagonist at dopamine D₂ receptors, induces autophagy by increasing reactive oxygen species in SH-SY5Y neuroblastoma cells.
Autophagy is associated with cell survival and cell death. Autophagy is implicated in the pathophysiology of various human diseases. In order to identify autophagy regulatory molecules, we screened a chemical drug library in SH-SY5Y cells and selected Sertindole as a potent autophagy inducer. Sertindole was developed as an antipsychotic drug for Schizophrenia. Sertindole treatment highly induced the formation of autophagosomes as well as LC3 conversion. Subsequently, Sertindole-induced autophagy was efficiently suppressed by down regulation of ATG5. Sertindole also increased reactive oxygen species (ROS) production, which contributes to autophagy-associated cell death in neuroblastoma cells. ROS scavengers such as N-acetylcysteine and Trolox suppressed not only ROS generation but also autophagy activation by Sertindole. These results suggest Sertindole induces autophagy and autophagy-associated cell death by ROS production in neuroblastoma cells.


Intracellular voltage of isolated epithelia of frog skin: apical and basolateral cell punctures.
Isolated epithelia of frog skin were prepared with collagenase, and the cells were punctured with intracellular microelectrodes across their apical (outer) and basolateral (inner) surfaces. Regardless of the route of cell puncture, the intracellular voltage (Vosc) in short-circuited isolated epithelia was markedly negative, averaging -70.4 mV for apical punctures and -91.6 mV for basolateral punctures. As in intact epithelia, amiloride outside caused the Vosc to become more negative (means of -96.7 and -101.8 mV), with a concomitant increase in the resistance of the apical barrier. Increasing the [K)i of the basolateral solution from 2.4 to 8.0 or 14.4 mM caused rapid step depolarization (5-10 s) of the Vosc under transepithelial Na transporting and amiloride-inhibited conditions of Na transport, with the delta Vosc ranging between 23.9 and 68.3 mV per decade change of [K]i. The finding that the Vosc of isolated epithelia of frog skin is independent of the route of cell penetration is consistent with the notion that the cells of the stratified epithelium are electrically coupled (functional syncitium). Moreover, the isolated epithelium can serve as a useful preparation, especially in studies designed to investigate the properties of the basolateral surfaces of cells.


Reproductive effort in viscous populations.
Here I study a kin selection model of reproductive effort, the allocation of resources to fecundity versus survival, in a patch-structured population. Breeding females remain in the same patch for life. Offspring have costly, partial long-distance dispersal and compete for breeding sites, which become vacant upon the death of previous occupants. The main result is that the evolutionarily stable reproductive effort decreases as offspring dispersal rate increases. The result can be understood as follows: In a well-mixed population with global competition, neither adults nor juveniles compete with relatives, but in a patch-structured population with dispersal restricted to the juvenile phase, juveniles experience relatively less competition with relatives than adults, thus making juveniles relatively more valuable. Because this asymmetry between adults and juveniles decreases with the dispersal rate, so does the evolutionarily stable level of allocation to fecundity.


Recurrent autonomic dysreflexia due to chronic aortic dissection in an adult male with cervical spinal cord injury.
Autonomic dysreflexia is a hypertensive clinical emergency for persons with spinal cord injury at T-6 level or above. Recurrent autonomic dysreflexia is uncommon in spinal cord injury patients and is usually caused by noxious stimuli that cannot be removed promptly, e.g., somatic pain, abdominal distension. A 61-year-old man, who sustained tetraplegia at C-5 (ASIA-A) 38 years ago, was admitted with chest infection. Computerised tomography (CT) of the chest showed the ascending aorta to measure 4 cm in anteroposterior diameter; descending thoracic aorta measured 3.5 cm. No dissection was seen. Normal appearances of abdominal aorta were seen. He was treated with noninvasive ventilation, antibiotics, and diuretics. Nineteen days later, when there was sudden deterioration in his clinical condition, CT of the pulmonary angiogram was performed to rule out pulmonary embolism. This showed no pulmonary embolus, but the upper abdominal aorta showed some dissection with thrombosis of the false lumen. Blood pressure was controlled with perindopril 2 mg, once a day, doxazosin 4 mg, twice a day, and furosemide 20 mg, twice a day. Since this patient did not show clinical features of mesenteric or lower limb ischaemia, the vascular surgeon did not recommend subdiaphragmatic aortic replacement. This patient subsequently developed recurrent episodes of autonomic dysreflexia. Each acute episode of dysreflexia was controlled by nifedipine given sublingually in doses varying from 5 to 20 mg. No inciting cause for autonomic dysreflexia could be found other than chronic aortic dissection. This patient's medication was then changed to doxazosin 8 mg, twice a day, and sustained-release nifedipine 10 mg, twice a day, which helped to prevent recurrent autonomic dysreflexia. Chronic aortic dissection is a very rare cause for recurrent autonomic dysreflexia in ageing spinal cord injury patients. When the inciting cause for dysreflexia is not amenable for treatment, recurrent dysreflexic episodes can be prevented by pharmacotherapy with an alpha-adrenergic blocking agent (doxazosin) and sustained-release nifedipine.


Expression and characterization of recombinant hepatitis A virus 3C proteinase.
The 3C proteinase from the hepatitis A virus (HAV) was cloned into a multicopy expression vector in Escherichia coli under control of the tac promoter. The resulting plasmid construction produced 3C proteinase as a soluble and active enzyme constituting approximately 10% of total cellular proteins. The enzyme was purified to apparent homogeneity as judged by SDS gel electrophoresis and HPLC reversed-phase and FPLC ion-exchange chromatography. A colorimetric assay was developed, and synthetic peptides derived from the predicted cleavage sites of the HAV polyprotein were tested for proteolysis of the enzyme. The peptide representing the 2B/2C cleavage site was cleaved most efficiently with a Km and kcat of 2.1 +/- 0.5 mM and 1.8 +/- 0.1 s-1, respectively. Site-directed mutagenesis was then used to identify the cysteine at position 172 as the active site nucleophile. Finally, the purified enzyme showed the expected endoproteinase activity on the P1 precursor protein generated by in vitro transcription/translation.


First-in-human study of SBRT and adenosine pathway blockade to potentiate the benefit of immunochemotherapy in early-stage luminal B breast cancer: results of the safety run-in phase of the Neo-CheckRay trial.
Luminal B breast cancer (BC) presents a worse prognosis when compared with luminal A BC and exhibits a lower sensitivity to chemotherapy and a lower immunogenicity in contrast to non-luminal BC subtypes. The Neo-CheckRay clinical trial investigates the use of stereotactic body radiation therapy (SBRT) directed to the primary tumor in combination with the adenosine pathway inhibitor oleclumab to improve the response to neo-adjuvant immuno-chemotherapy in luminal B BC. The trial consists of a safety run-in followed by a randomized phase II trial. Here, we present the results of the first-in-human safety run-in. The safety run-in was an open-label, single-arm trial in which six patients with early-stage luminal B BC received the following neo-adjuvant regimen: paclitaxel q1w×12 → doxorubicin/cyclophosphamide q2w×4; durvalumab (anti-programmed cell death receptor ligand 1 (PD-L1)) q4w×5; oleclumab (anti-CD73) q2w×4 → q4w×3 and 3×8 Gy SBRT to the primary tumor at week 5. Surgery must be performed 2-6 weeks after primary systemic treatment and adjuvant therapy was given per local guidelines, RT boost to the tumor bed was not allowed. Key inclusion criteria were: luminal BC, Ki67≥15% or histological grade 3, MammaPrint high risk, tumor size≥1.5 cm. Primary tumor tissue samples were collected at three timepoints: baseline, 1 week after SBRT and at surgery. Tumor-infiltrating lymphocytes, PD-L1 and CD73 were evaluated at each timepoint, and residual cancer burden (RCB) was calculated at surgery. Six patients were included between November 2019 and March 2020. Median age was 53 years, range 37-69. All patients received SBRT and underwent surgery 2-4 weeks after the last treatment. After a median follow-up time of 2 years after surgery, one grade 3 adverse event (AE) was reported: pericarditis with rapid resolution under corticosteroids. No grade 4-5 AE were documented. Overall cosmetical breast evaluation after surgery was 'excellent' in four patients and 'good' in two patients. RCB results were 2/6 RCB 0; 2/6 RCB 1; 1/6 RCB 2 and 1/6 RCB 3. This novel treatment combination was considered safe and is worth further investigation in a randomized phase II trial. NCT03875573.


Store-operated Ca2+ entry supports contractile function in hearts of hibernators.
Hibernators have a distinctive ability to adapt to seasonal changes of body temperature in a range between 37°C and near freezing, exhibiting, among other features, a unique reversibility of cardiac contractility. The adaptation of myocardial contractility in hibernation state relies on alterations of excitation contraction coupling, which becomes less-dependent from extracellular Ca2+ entry and is predominantly controlled by Ca2+ release from sarcoplasmic reticulum, replenished by the Ca2+-ATPase (SERCA). We found that the specific SERCA inhibitor cyclopiazonic acid (CPA), in contrast to its effect in papillary muscles (PM) from rat hearts, did not reduce but rather potentiated contractility of PM from hibernating ground squirrels (GS). In GS ventricles we identified drastically elevated, compared to rats, expression of Orai1, Stim1 and Trpc1/3/4/5/6/7 mRNAs, putative components of store operated Ca2+ channels (SOC). Trpc3 protein levels were found increased in winter compared to summer GS, yet levels of Trpc5, Trpc6 or Trpc7 remained unchanged. Under suppressed voltage-dependent K+, Na+ and Ca2+ currents, the SOC inhibitor 2-aminoethyl diphenylborinate (2-APB) diminished whole-cell membrane currents in isolated cardiomyocytes from hibernating GS, but not from rats. During cooling-reheating cycles (30°C-7°C-30°C) of ground squirrel PM, 2-APB did not affect typical CPA-sensitive elevation of contractile force at low temperatures, but precluded the contractility at 30°C before and after the cooling. Wash-out of 2-APB reversed PM contractility to control values. Thus, we suggest that SOC play a pivotal role in governing the ability of hibernator hearts to maintain their function during the transition in and out of hibernating states.


Interleukin-1beta and barrier function of retinal pigment epithelial cells (ARPE-19): aberrant expression of junctional complex molecules.
To examine the effects of interleukin (IL)-1beta on the resistance and permeability of the tight junctions of cultured retinal pigment epithelial cells. A human RPE cell line (ARPE-19) cultured on microporous filter-supports was used. IL-1beta and monoclonal anti-IL-1beta antibody-treated IL-1beta (mAbIL-1beta) were added to the standard culture medium. Transepithelial resistance (TER) of confluent RPE cells was measured by epithelial voltmeter. The permeability of the RPE cells to sodium fluorescein, horseradish peroxidase, and inulin was measured. The expression of the occludin and claudin was determined by real-time polymerase chain reaction (PCR), immunohistochemistry, and Western blot analysis. A significantly greater decrease of TER occurred in IL-1beta-supplemented medium than in standard medium plus mAbIL-1beta after several days of stimulation. A significantly greater increase of sodium fluorescein, horseradish peroxidase, and inulin permeability occurred in IL-1beta-supplemented medium than in standard medium. The expression of the occludin gene and some types of claudin genes was observed. The expression of occludin was downregulated and that of claudin-1 upregulated more in IL-1beta-supplemented medium than in standard medium by real-time PCR, immunohistochemistry, and Western blot analysis. The tight junctions of ARPE-19 cells are altered by IL-1beta supplementation either directly or through other factors activated by IL-1beta. The downregulation of occludin and upregulation of claudin-1 may have participated in the dysfunction of the RPE tight junctions in these in vitro experiments.


Heterotetracenes: Flexible Synthesis and in Silico Assessment of the Hole-Transport Properties.
Thienoacenes and furoacenes are among the most frequent molecular units found in organic materials. The efficient synthesis of morphologically different heteroacenes and the rapid determination of their solid-state and electronic properties are still challenging tasks, which slow down progress in the development of new materials. Here, we report a flexible and efficient synthesis of unprecedented heterotetracenes based on a platinum- and gold-catalyzed cyclization-alkynylation domino process using EthynylBenziodoXole (EBX) hypervalent iodine reagents in the key step. The proof-of-principle in silico estimation of the synthesized tetracenes' charge transport properties reveals their strong dependence on both the position and nature of the heteroatoms in the ring system. A broad range of mobility is predicted, with some compounds displaying performance potentially comparable to that of state-of-the-art electronic organic materials.


Evaluation of PCR in detection of Mycobacterium tuberculosis from formalin-fixed, paraffin-embedded tissues: comparison of four amplification assays.
We compared the sensitivities and specificities of four nested PCR assays for the detection of Mycobacterium tuberculosis from formalin-fixed, paraffin-embedded tissues. Thirty-seven autopsy samples from human immunodeficiency virus-positive patients were analyzed: 15 were M. tuberculosis positive, 11 served as negative controls, and 11 were Ziehl-Neelsen positive without cultural confirmation of M. tuberculosis. Three genomic sequences (mtp40, 65-kDa antigen gene, and IS6110) with different molecular masses and numbers of repetitions within the M. tuberculosis genome were targeted. On the IS6110 sequence, two fragments of different sizes (106 and 123 bp, respectively) were amplified with two separate pairs of primers. The highest sensitivity rates were obtained by amplifying the highly repetitive IS6110 insertion sequence, and the different primers tested showed a sensitivity ranging from 80 to 87%. Amplification of the large 223-bp fragment of the mtp40 sequence present in a single copy in the M. tuberculosis genome yielded a high rate of false-negative results, ranging from 66 to 80%. A poor sensitivity (from 47 to 60%) was also shown by PCR amplification of the 142-bp 65-kDa antigen gene. All the PCRs except that for the 65-kDa antigen gene showed a specificity of 100%. Moreover, different results were obtained with different dilutions of DNA, and DNA concentrations of 1 and 3 microg yielded the highest sensitivities depending upon which protocol was used. Application of the PCRs to the Ziehl-Neelsen-positive, culture-negative samples confirmed the sensitivities of the PCRs obtained with the control samples. In conclusion, PCR can successfully be used to detect M. tuberculosis from paraffin-embedded tissues and can be particularly useful in the validation of a diagnosis of tuberculosis in clinical settings in which the diagnosis is uncertain. However, the efficacy of PCR strictly depends on several amplification parameters such as DNA concentration, target DNA size, and the repetitiveness of the amplified sequence.


Relations of photometry. Part 2.
The most general photometric definitions are presented and the origins of Lamberts's law and the cosine (or inverse square) law of illumination are shown. These derivations are made possible through the use of a simple diagram relating emitting and illuminated surfaces. Several classical problems are solved to illustrate the theory and use of the diagram, and the problems of the photometric microscope and telescope are discussed in detail.


Two-transcript gene expression classifiers in the diagnosis and prognosis of human diseases.
Identification of molecular classifiers from genome-wide gene expression analysis is an important practice for the investigation of biological systems in the post-genomic era--and one with great potential for near-term clinical impact. The 'Top-Scoring Pair' (TSP) classification method identifies pairs of genes whose relative expression correlates strongly with phenotype. In this study, we sought to assess the effectiveness of the TSP approach in the identification of diagnostic classifiers for a number of human diseases including bacterial and viral infection, cardiomyopathy, diabetes, Crohn's disease, and transformed ulcerative colitis. We examined transcriptional profiles from both solid tissues and blood-borne leukocytes. The algorithm identified multiple predictive gene pairs for each phenotype, with cross-validation accuracy ranging from 70 to nearly 100 percent, and high sensitivity and specificity observed in most classification tasks. Performance compared favourably with that of pre-existing transcription-based classifiers, and in some cases was comparable to the accuracy of current clinical diagnostic procedures. Several diseases of solid tissues could be reliably diagnosed through classifiers based on the blood-borne leukocyte transcriptome. The TSP classifier thus represents a simple yet robust method to differentiate between diverse phenotypic states based on gene expression profiles. Two-transcript classifiers have the potential to reliably classify diverse human diseases, through analysis of both local diseased tissue and the immunological response assayed through blood-borne leukocytes. The experimental simplicity of this method results in measurements that can be easily translated to clinical practice.


Climate change and the characterization, breeding and conservation of animal genetic resources.
Livestock production both contributes to and is affected by climate change. In addition to the physiological effects of higher temperatures on individual animals, the consequences of climate change are likely to include increased risk that geographically restricted rare breed populations will be badly affected by disturbances. Indirect effects may be felt via ecosystem changes that alter the distribution of animal diseases or affect the supply of feed. Breeding goals may have to be adjusted to account for higher temperatures, lower quality diets and greater disease challenge. Species and breeds that are well adapted to such conditions may become more widely used. Climate change mitigation strategies, in combination with ever increasing demand for food, may also have an impact on breed and species utilization, driving a shift towards monogastrics and breeds that are efficient converters of feed into meat, milk and eggs. This may lead to the neglect of the adaptation potential of local breeds in developing countries. Given the potential for significant future changes in production conditions and in the objectives of livestock production, it is essential that the value provided by animal genetic diversity is secured. This requires better characterization of breeds, production environments and associated knowledge; the compilation of more complete breed inventories; improved mechanisms to monitor and respond to threats to genetic diversity; more effective in situ and ex situ conservation measures; genetic improvement programmes targeting adaptive traits in high-output and performance traits in locally adapted breeds; increased support for developing countries in their management of animal genetic resources; and wider access to genetic resources and associated knowledge.


Mental health, suicide attempt, and family function for adolescents' primary health care during the COVID-19 pandemic.
Background: The study's purpose was to identify associations between mental health risk, suicide attempts, and family function. Methods: A correlational, descriptive, and cross-sectional study was carried out in a group of adolescents in the last grade of secondary school to establish the association between mental health risk, suicide attempt, and family functionality. The instruments used were the self-report questionnaire, the suicide risk assessment scale, and the family APGAR. Data analysis was performed using the artificial intelligence algorithm (gower clustering). Results: 246 adolescents responded to the three instruments, which made it possible to select those with correlations of sensitive interest and, based on these, an intervention plan. Psychological distress was found in 28%, psychotic symptoms in 85%, and problematic alcohol use in 9%. Good family functioning was identified in 34% and some type of family dysfunction in 66%. In terms of suicide risk, there was a low suicide risk of 74%, 24% medium risk, and 2% high risk. It could be shown that there is a correlation in a group of 15% of the respondents. Conclusions: The risk of suffering mental health deterioration and the suicide risk, during this pandemic period, seems to be related to family functionality.


Streptomyces Dominate the Soil Under Betula Trees That Have Naturally Colonized a Red Gypsum Landfill.
The successful restoration of well-engineered tailings storage facilities is needed to avoid mine tailings problems. This study characterized the bacterial communities from vegetated and non-vegetated soils from a red gypsum landfill resulting from the industrial extraction of titanium. A set of 275 bacteria was isolated from vegetated soil and non-vegetated soil areas and taxonomically characterized using BOX-PCR. The study also evaluated the ability of a subset of 88 isolated bacteria on their ability to produce plant growth promoting (PGP) traits [indoleacetic acid (IAA) production, phosphate solubilization, and siderophore production] and their tolerance to potentially toxic elements (PTEs). Twenty strains were chosen for further analysis to produce inoculum for birch-challenging experiments. Principal component analysis (PCA) showed that the set of pedological parameters (pH, granulometry, carbon, organic matter, and Mg content) alone explained approximately 40% of the differences between the two soils. The highest density of total culturable bacteria was found in the vegetated soil, and it was much higher than that in the non-vegetated soil. The Actinobacteria phyla dominated the culturable soil community (70% in vegetated soil and 95% in non-vegetated soil), while the phyla Firmicutes (including the genus Bacillus) and Bacteroides (including the genera Pedobacter and Olivibacter) were found only in the vegetated soil fraction. Additional genera (Rhizobium, Variovorax, and Ensifer) were found solely in the vegetated soil. The vegetated soil bacteria harbored the most beneficial PGP bacteria with 12% of the isolates showing three or more PGP traits. The strains with higher metal tolerances in our study were Phyllobacterium sp. WR140 (RO1.15), Phyllobacterium sp. WR140 (R01.34), and Streptomyces sp. (R04.15), all isolated from the vegetated soil. Among the isolates tested in challenging experiments, Phyllobacterium (R01.34) and Streptomyces sp. (R05.33) have the greatest potential to act as PGP rhizobacteria and therefore to be used in the biological restoration of tailings dumps.


A parallelogram metallomacrocycle bearing self-catenation and its derivative supramolecular isomerism.
Macrocycle-based architectures are of importance in synthetic chemistry. Here, a novel parallelogram metallomacrocycle Fe2(LA)2 with reversible structural transformation was designed and synthesized. The template-free metalla[2]catenane [Fe2(LA)2]2 could be obtained by changing the concentration and has been monitored on the basis of NMR analysis. By redesigning the metallo-ligand, a catenane-like intersected parallelogram assembly with two shape-persistent supramolecular isomers was achieved. This work develops the field of terpyridine-based macrocycle system research and is valuable for obtaining other types of supramolecular isomerism.


Awareness about Tobacco Causing Head and Neck Cancers via Mass Media: A Case-Control Study from India.
This study aimed to assess the awareness of people about the adverse effects of tobacco (smoking and chewing) consumption causing head and neck cancers (HNCs) via mass media channels like television, cinema, radio and newspapers or magazines, wall painting or billboards / hoardings, public transportation and packets of chewing tobacco, bidis or cigarettes. Hospital-based case-control was conducted in Pune, Maharashtra, India. Face to face interviews were conducted for the purpose of data collection on 225 cases and 240 controls. The relationship between two categorical variables were estimated using chi-square test with a 2-tailed P value of <.05. SPSS software was used for data analysis. Controls as compared to cases had good awareness scores for chewing (59.9%) and smoking tobacco (63.7%), P<0.001. The most common form of mass media was television where the cases (60.4%) and controls (77.9%) had heard messages about tobacco in chewing and smoking form causing HNCs. Level of awareness of tobacco causing HNCs amongst tobacco users, stratified by their status (cases versus controls) showed that cases were 1.68 times less likely than controls to have heard or seen messages about the association between chewing tobacco and HNCs via radio. Males (61.3% and 61.0%) had significantly (P<0.001) more awareness as compared to females (46.9% and 43.5%) about chewing and smoking tobacco as a causal factor for HNCs. Mass media needs to create a social environment which discourages tobacco consumption and promotes oral health at the population level. Additionally, there should be easy access to the availability of support services like Quitline and other community support services.


Colon and rectal cancer incidence and water trihalomethane concentrations in New South Wales, Australia.
There is evidence, although inconsistent, that long term exposure to disinfection by products (DBPs) increases the risk of bowel cancer. No study has been conducted in Australia to examine this association and due to difference in the methods of disinfection the risk can vary across geographical regions and. This study was conducted to analyse the association of trihalomethanes (THMs) in water with colon and rectal cancer in NSW Australia. Average yearly concentrations of total and individual species of THMs were obtained for 50 local government areas (LGAs). Indirectly-standardized incidence rates of colon and rectal cancers in LGAs for the period 1995 to 2001 were regressed against mean THM concentrations lagged five years, adjusting for socioeconomic status, high risk drinking, smoking status, usual source of water and year of diagnosis, including local and global random effects within a Bayesian framework. The incidence rate ratios (IRRs) for an interquartile range (IQR) increase in THMs were estimated. Using five year lag of exposure there was a positive association between bromoform concentration and CRC in men (IRR = 1.025, 95% CI 1.010, 1.040) but not in women (IRR = 1.003, 95% CI 0.987, 1.018). The association in men was mainly found in colon cancer with bromoform (IRR = 1.035, 95% CI 1.017, 1.053). There was no appreciable association of colorectal cancer with other species of THMs. Sensitivity analyses did not materially change the associations observed. A positive association was observed between colon cancer and water bromoform concentrations in men. Given the potential population impact of such an association, further research into the relationship between THMs, particularly brominated species, and colorectal cancer is warranted.


A. Conan Doyle, Sherlock Holmes, and murder by tropical infection.
The scientific insights with which A. Conan Doyle endowed his creation, the master detective Sherlock Holmes, continue to attract scholarly interest. Indeed, the clinical and/or scientific aspects of Doyle's fiction hold appeal for those interested in the epidemiology of tropical infectious diseases. The origins and routes of transmission of tropical infections were subjects of fruitful investigation in the latter half of the nineteenth century. In "The Adventure of the Dying Detective," Sherlock Holmes investigates a murder that he suspects to have resulted from a fatal Asiatic disease associated with a short incubation period: the indications point to primary septicemic plague as the murder weapon.


Oral inflammatory process and general health. Part 2: How does the periapical inflammatory process compromise general health?
At present, the focal infection theory still has very controversial aspects. In spite of the great number of studies, there is no evidence that focal infections or even antigenic mimicry are responsible for anything other than sporadic abscesses/infections and possibly rare autoimmune disorders. linflammation of endodontic origin (i.e., apical periodontitis--AP) has not received the same attention as inflammation originating from the periodontium. Endodontics is a microbiological problem, since the bacterial infection is the "prime mover" of pulp (before) and periapical (later) disease. The aims of endodontic treatment have to be considered from a microbiological viewpoint. Considering these problems in this second part of their study, the Authors, after close examination of the virulence of microorganisms and of the host defense, analyze the endodontic infection and microbiological species. They emphasize the possibility of a relationship between periapical inflammatory lesions and bacterial endocarditis in preventing metafocal disease. Bacterial endocarditis deserves special mention because despite involving specialists of two scientific fields, its prophylaxis is almost always assigned to medical practice, and especially, to dentistry. Given the dangers of the disease, antibiotic prophylaxis is both absolutely necessary and can be very effective, and it should be used especially in clinical situations with high risk individuals. However, the ability of antibiotic therapy to prevent or reduce the frequency, magnitude or duration of bacteremia associated with a dental procedure is controversial. Studies should also be undertaken to determine to compare the efficacy of endodontic treatment with alternative therapy such as implants, prosthetic replacements or no treatment other than extraction. To date, these studies have not been carried out, and there is no evidence to support the theory that modern endodontic therapy is not safe and effective.


Shift in cytotype frequency and niche space in the invasive plant Centaurea maculosa.
Polyploidy is often assumed to increase the spread and thus the success of alien plant species, but few empirical studies exist. We tested this hypothesis with Centaurea maculosa Lam., a species native to Europe and introduced into North America approximately 120 years ago where it became highly invasive. We analyzed the ploidy level of more than 2000 plants from 93 native and 48 invasive C. maculosa populations and found a pronounced shift in the relative frequency of diploid and tetraploid cytotypes. In Europe diploid populations occur in higher frequencies than tetraploids and only four populations had both cytotypes, while in North America diploid plants were found in only one mixed population and thus tetraploids clearly dominated. Our results showed a pronounced shift in the climatic niche between tetraploid populations in the native and introduced range toward drier climate in North America and a similar albeit smaller shift between diploids and tetraploids in the native range. The field data indicate that diploids have a predominately monocarpic life cycle, while tetraploids are often polycarpic. Additionally, the polycarpic life-form seems to be more prevalent among tetraploids in the introduced range than among tetraploids in the native range. Our study suggests that both ploidy types of C. maculosa were introduced into North America, but tetraploids became the dominant cytotype with invasion. We suggest that the invasive success of C. maculosa is partly due to preadaptation of the tetraploid cytotype in Europe to drier climate and possibly further adaptation to these conditions in the introduced range. The potential for earlier and longer seed production associated with the polycarpic life cycle constitutes an additional factor that may have led to the dominance of tetraploids over diploids in the introduced range.


Prophylactic endoscopic sclerotherapy of oesophageal varices in liver cirrhosis. A multicentre prospective controlled randomised trial in Vienna.
The effect of prophylactic treatment of oesophageal varices by endoscopic injection sclerotherapy before the first episode of variceal haemorrhage was studied in patients with cirrhosis in a prospective, randomised and controlled multicentre trial. From February 1984 to March 1987 patients with liver cirrhosis and large varices (stage III-IV according to Paquet) were treated and followed up. The sample comprised 87 patients: 45 in the prophylactic treatment and 42 in the control group. After excluding drop outs, 41 patients were treated in each group. Twenty nine per cent of patients in the sclerotherapy group and 34% in the control group had a variceal haemorrhage during the period of observation. There was no significant difference in the distributions of the bleeding free intervals between the sclerotherapy and the control groups. During the follow up period 24% of patients in the sclerotherapy group and 46% in the control group died. The distribution of survival times indicates a tendency towards longer survival of patients with prophylactic sclerotherapy, particularly in those with alcoholic cirrhosis.


[The correlation between the heart contraction rate and the operator's stress-resistance].
There is factual confirmation of the correlation between operator's stress-resistance and heart rate (HR). HR data alone without assessment of the job quality in a group of operators were enough to rank the operators by their resistance to stress. However, there are several requirements to be met. To begin with, operators should experience a weak or moderate mental stress that will modify HR. Secondly, other sympathetic influences should be reduced to a minimum, as they may mask the emotion-driven effects (motor activity, elevated tone of posture muscles, meals). With this in view, HR measurements are advisable during night sleep to escape, in addition, the endogenous sympathetic activation associated with ergotrophic metabolism. In the day time, HR should be measured after 10 to 15 minutes in recumbent position.


Mobile healthcare for automatic driving sleep-onset detection using wavelet-based EEG and respiration signals.
Driving drowsiness is a major cause of traffic accidents worldwide and has drawn the attention of researchers in recent decades. This paper presents an application for in-vehicle non-intrusive mobile-device-based automatic detection of driver sleep-onset in real time. The proposed application classifies the driving mental fatigue condition by analyzing the electroencephalogram (EEG) and respiration signals of a driver in the time and frequency domains. Our concept is heavily reliant on mobile technology, particularly remote physiological monitoring using Bluetooth. Respiratory events are gathered, and eight-channel EEG readings are captured from the frontal, central, and parietal (Fpz-Cz, Pz-Oz) regions. EEGs are preprocessed with a Butterworth bandpass filter, and features are subsequently extracted from the filtered EEG signals by employing the wavelet-packet-transform (WPT) method to categorize the signals into four frequency bands: α, β, θ, and δ. A mutual information (MI) technique selects the most descriptive features for further classification. The reduction in the number of prominent features improves the sleep-onset classification speed in the support vector machine (SVM) and results in a high sleep-onset recognition rate. Test results reveal that the combined use of the EEG and respiration signals results in 98.6% recognition accuracy. Our proposed application explores the possibility of processing long-term multi-channel signals.


Delusions with religious content in patients with psychosis: how they interact with spiritual coping.
Delusions with religious content have been associated with a poorer prognosis in schizophrenia. Nevertheless, positive religious coping is frequent among this population and is associated with a better outcome. The aim of this study was to compared patients with delusions with religious content (n = 38), patients with other sorts of delusions (n = 85) and patients without persistent positive symptoms (n = 113) clinically and spiritually. Outpatients (n = 236) were randomly selected for a quantitative and qualitative evaluation of religious coping. Patients presenting delusions with religious content were not associated with a more severe clinical status compared to other deluded patients, but they were less likely to adhere to psychiatric treatment. For almost half of the group (45%), spirituality and religiousness helped patients cope with their illness. Delusional themes consisted of: persecution (by malevolent spiritual entities), influence (being controlled by spiritual entities), and self-significance (delusions of sin/guilt or grandiose delusions). Both groups of deluded patients valued religion more than other patients, but patients presenting delusions with religious content received less support from religious communities. In treating patients with such symptoms, clinicians should go beyond the label of "religious delusion," likely to involve stigmatization, by considering how delusions interact with patients' clinical and psychosocial context.


Tailgut cyst: diagnosis with CT and sonography.
Tailgut cyst is a rare congenital lesion that presents as a presacral mass, found in infancy or persisting undetected into adulthood, and characterized by multiple cysts lined with a gastrointestinal type of epithelium. In a retrospective review of five patients whose ages ranged from 1 month to 42 years--three cases with CT and two with sonograms--tailgut cyst appeared as a complex mass on both procedures, representing its composition of multiple small cysts filled with keratinous material and other debris. Other CT and sonographic features include well-defined borders, and, in one case, presentation as a mass lateral to the midline. CT also shows preservation of adjacent fat planes. Tailgut cyst should be included in the differential diagnosis of a retrorectal mass at any age.


Combined intraoperative and postoperative irradiation for soft tissue sarcomas.
A major problem in the management of soft tissue sarcomas is the high rate of local recurrence postoperatively, regardless of whether the surgery performed was radical or conservative. The addition of irradiation to the tumor bed has decreased this risk significantly. In an attempt to achieve maximal local tumor control, a number of patients were treated by means of excisional surgery combined with an intraoperative implant and postoperative external irradiation. A new type of intraoperative irradiation procedure was used, in which iridium (192Ir) is inserted into afterloading nylon catheters which are implanted in the tumor bed at the time that the tumor is surgically excised.


Autologous transfusion of stored red blood cells increases pulmonary artery pressure.
Transfusion of erythrocytes stored for prolonged periods is associated with increased mortality. Erythrocytes undergo hemolysis during storage and after transfusion. Plasma hemoglobin scavenges endogenous nitric oxide leading to systemic and pulmonary vasoconstriction. We hypothesized that transfusion of autologous blood stored for 40 days would increase the pulmonary artery pressure in volunteers with endothelial dysfunction (impaired endothelial production of nitric oxide). We also tested whether breathing nitric oxide before and during transfusion could prevent the increase of pulmonary artery pressure. Fourteen obese adults with endothelial dysfunction were enrolled in a randomized crossover study of transfusing autologous, leukoreduced blood stored for either 3 or 40 days. Volunteers were transfused with 3-day blood, 40-day blood, and 40-day blood while breathing 80 ppm nitric oxide. The age of volunteers was 41 ± 4 years (mean ± SEM), and their body mass index was 33.4 ± 1.3 kg/m(2). Plasma hemoglobin concentrations increased after transfusion with 40-day and 40-day plus nitric oxide blood but not after transfusing 3-day blood. Mean pulmonary artery pressure, estimated by transthoracic echocardiography, increased after transfusing 40-day blood (18 ± 2 to 23 ± 2 mm Hg; P < 0.05) but did not change after transfusing 3-day blood (17 ± 2 to 18 ± 2 mm Hg; P = 0.5). Breathing nitric oxide decreased pulmonary artery pressure in volunteers transfused with 40-day blood (17 ± 2 to 12 ± 1 mm Hg; P < 0.05). Transfusion of autologous leukoreduced blood stored for 40 days was associated with increased plasma hemoglobin levels and increased pulmonary artery pressure. Breathing nitric oxide prevents the increase of pulmonary artery pressure produced by transfusing stored blood. Clinical trial registered with www.clinicaltrials.gov (NCT 01529502).


Long-term behavioral and biochemical effects of an ultra-low dose of Δ9-tetrahydrocannabinol (THC): neuroprotection and ERK signaling.
We have previously reported that a single injection of an ultra-low dose of delta-9-tetrahydrocannabinol (THC; the psychoactive ingredient of marijuana) protected the brain from pentylenentetrazole (PTZ)-induced cognitive deficits when applied 1-7 days before or 1-3 days after the insult. In the present study we expanded the protective profile of THC by showing that it protected mice from cognitive deficits that were induced by a variety of other neuronal insults, including pentobarbital-induced deep anesthesia, repeated treatment with 3,4 methylenedioxymethamphetamine (MDMA; "ecstasy") and exposure to carbon monoxide. The protective effect of THC lasted for at least 7 weeks. The same ultra-low dose of THC (0.002 mg/kg, a dose that is 3-4 orders of magnitude lower than the doses that produce the known acute effects of the drug in mice) induced long-lasting (7 weeks) modifications of extracellular signal-regulated kinase (ERK) activity in the hippocampus, frontal cortex and cerebellum of the mice. The alterations in ERK activity paralleled changes in its activating enzyme MEK and its inactivating enzyme MKP-1. Furthermore, a single treatment with the low dose of THC elevated the level of pCREB (phosphorylated cAMP response element-binding protein) in the hippocampus and the level of BDNF (brain-derived neurotrophic factor) in the frontal cortex. These long-lasting effects indicate that a single treatment with an ultra-low dose of THC can modify brain plasticity and induce long-term behavioral and developmental effects in the brain.


Comparison of phenotypic versus marker-assisted background selection for the SUB1 QTL during backcrossing in rice.
Marker assisted backcrossing has been used effectively to transfer the submergence tolerance gene SUB1 into popular rice varieties, but the approach can be costly. The selection strategy comprising foreground marker and phenotypic selection was investigated as an alternative. The non-significant correlation coefficients between ranking of phenotypic selection and ranking of background marker selection in BC(2)F(1), BC(3)F(1) and BC(3)F(2) generations indicated inefficiency of phenotypic selection compared to marker-assisted background selection with respect to recovery of the recipient genome. In addition, the introgression size of the chromosome fragment containing SUB1 was approximately 17 Mb, showing the effects of linkage drag. The significant correlation coefficient between rankings of phenotypic selection with the percentage of recipient alleles in the BC(1)F(1) generation suggested that background selection could be avoided in this generation to minimize the genotyping cost. The phenotypically selected best plant of the BC(3)F(1) generation was selfed and backcross recombinant lines were selected in the resulting BC(3)F(4) generation. The selection strategy could be appropriate for the introgression of SUB1 QTL in countries that lack access to high-throughput genotyping facilities.


Surgeons' attitudes toward HIV/AIDS in Turkey.
Currently HIV/AIDS is one of the most difficult challenges for health care professionals. As primary medical care providers, attitudes of the physicians towards HIV/AIDS are of utmost significance. The aim of this research is to determine the attitudes of the surgeons towards HIV/AIDS in their working environment. Data were collected from a self-administered questionnaire given to 128 surgeons. Results revealed that gender, age and professional experience are not significant. This study demonstrated that doctors are worried about contracting HIV/AIDS from the patients. In some circumstances doctors' attitudes, are not clear which may lead to some ethical problems. It is concluded that doctors overestimate the risks and they need special education about HIV/AIDS, as well as professional help to handle their attitudes toward HIV/AIDS.


Mandibular overdentures retained by two implants: 10-year results from a crossover clinical trial comparing ball-socket and bar-clip attachments.
The aim of this study was to evaluate patient satisfaction and clinical and prosthetic outcomes of two-implant mandibular overdenture treatment with different attachment types after 10 years of function. In a crossover clinical trial, 18 edentulous subjects with complaints regarding their mandibular dentures received two implants and a new denture with magnet, ball-socket, or bar-clip attachments that were applied in a random order. At the end of the experiment, the attachment type of their choosing was fitted in the overdenture. After 10 years, 7 subjects with a ball-socket and 7 subjects with a bar-clip attachment were available for evaluation. The same questionnaire from 10 years before was completed, and subjects were asked to express their overall appreciation of their dentures on a visual analog scale (VAS). Six scales of denture complaints were constructed. Mean scale and VAS scores between initial evaluation and after 10 years were compared. In addition, marginal probing depths, Bleeding Index, and radiographic marginal bone loss were assessed. There was no marked difference in satisfaction between subjects with ball-socket- and bar-clip-retained two-implant mandibular overdentures at initial evaluation and after 10 years of function. Patients' appreciation of their implant-retained denture was and remained high over time. Clinical parameters revealed healthy mucosal conditions and stable marginal bone levels, determined radiographically. Probing depths around implants provided with ball-socket attachments were slightly shallower than those with bar-clip attachments after 10 years of function (P < .05).


An Integrated Mechanistic Model of Mindfulness-Oriented Recovery Enhancement for Opioid-Exposed Mother-Infant Dyads.
A growing body of neurobiological and psychological research sheds light on the mechanisms underlying the development and maintenance of opioid use disorder and its relation to parenting behavior. Perinatal opioid use is associated with risks for women and children, including increased risk of child maltreatment. Drawing from extant data, here we provide an integrated mechanistic model of perinatal opioid use, parenting behavior, infant attachment, and child well-being to inform the development and adaptation of behavioral interventions for opioid-exposed mother-infant dyads. The model posits that recurrent perinatal opioid use may lead to increased stress sensitivity and reward dysregulation for some mothers, resulting in decreased perceived salience of infant cues, disengaged parenting behavior, disrupted infant attachment, and decreased child well-being. We conclude with a discussion of Mindfulness-Oriented Recovery Enhancement as a means of addressing mechanisms undergirding perinatal opioid use, parenting, and attachment, presenting evidence on the efficacy and therapeutic mechanisms of mindfulness. As perinatal opioid use increases in the United States, empirically informed models can be used to guide treatment development research and address this growing concern.


Stem cells: a promising source for vascular regenerative medicine.
The rising and diversity of many human vascular diseases pose urgent needs for the development of novel therapeutics. Stem cell therapy represents a challenge in the medicine of the twenty-first century, an area where tissue engineering and regenerative medicine gather to provide promising treatments for a wide variety of diseases. Indeed, with their extensive regeneration potential and functional multilineage differentiation capacity, stem cells are now highlighted as promising cell sources for regenerative medicine. Their multilineage differentiation involves environmental factors such as biochemical, extracellular matrix coating, oxygen tension, and mechanical forces. In this review, we will focus on human stem cell sources and their applications in vascular regeneration. We will also discuss the different strategies used for their differentiation into both mature and functional smooth muscle and endothelial cells.


Effects of the novel coronary selective calcium channel blocker, McN-6186, on cardiocirculatory dynamics, coronary vascular resistance, and cardiac output distribution in normal, conscious rats.
The purpose of this study was to characterize the cardiocirculatory effects of the novel calcium channel blocker, McN-6186 (McN), normal, conscious rats. Animals were instrumented under halothane anesthesia for right atrial, left ventricular, arterial, and venous pressure recordings. The radioactive microsphere technique was used to measure regional blood flow (RBF) and cardiac output (CO) before (control) and during intravenous (i.v.) infusion of either McN at three doses (0.03, 0.1, 0.3 mg/kg) or vehicle at an equal infusion rate (0.0408 ml/min). Nifedipine (NIF) was also studied at three similar blood pressure (BP)-lowering doses (0.025, 0.150, and 0.375 mg/kg). The predominant effect of McN in conscious rats was to cause coronary vasodilation. The coronary vasodilator potency of McN was similar to NIF (ED25, McN = 0.03 mg/kg, NIF = 0.025 mg/kg). Neither McN nor NIF significantly changed systemic vascular resistance (SVR) over their respective coronary vasodilator dose ranges, suggesting that both compounds are selective coronary vasodilators. The doses of McN and NIF that reduced mean arterial pressure (MAP) by 25% (ED25) were similar (McN = 0.3 mg/kg, NIF = 0.375 mg/kg). At equal BP-lowering doses, McN increased coronary flow by 145% versus 110% for NIF. McN did not have major effects on other regions of the peripheral circulation. There was, however, some vasodilator activity in the renal and cerebral vascular beds. Because McN reduced coronary vascular resistance at a dose lower than that required to reduce resistance in other vascular beds, this compound appears to be a selective coronary vasodilator and may have therapeutic efficacy as an antianginal agent.


Human immunodeficiency and hepatitis virus infections and their associated conditions and treatments among people with haemophilia.
Treatment with contaminated plasma products before 1990 resulted in extraordinary prevalence rates of human immunodeficiency virus (HIV) and hepatitis B and C viruses (HBV, HCV). In the Second Multicentre Haemophilia Cohort Study (MHCS-II) during 2001-03, 30% of HCV-seropositive survivors had HIV and 4.6% were HBV carriers. Highly active antiretroviral therapy (HAART) radically altered the consequences of HIV/HCV coinfection. Whereas opportunistic infections predominated previously, current major complications are liver failure and bleeding (exacerbated by decreased clotting factor synthesis, hypersplenic thrombocytopenia, and oesophageal varices). Most HIV-positives in MHCS-II were HIV RNA-negative and had > 200 CD4(+) cells microL(-1), but only 59% were on HAART. With HIV, especially after 41 years of age, liver disease was apparent (jaundice in 5%, ascites 7%, hepatomegaly 9%, splenomegaly 19%). HAART increases survival but may contribute to various comorbidities. Without HIV, sustained HCV clearance is obtained in > 50% with combined pegylated interferons plus ribavirin, but data in haemophilic populations, especially with HIV, are limited. In MHCS-II, HCV RNA negativity was 41% following standard interferon plus ribavirin; among interferon-naive participants (implying spontaneous HCV clearance), HCV RNA negativity was 12% with and 25% without HIV. Without HIV, spontaneous HCV clearance was much more likely with early age at infection and particularly with recent birth (late 1970s or early 1980s) but not with bleeding propensity or its treatment. Most (72%) participants had received no anti-HCV therapy. Hepatic and haematological conditions are likely to increase during the coming years unless most adult haemophiliacs are successfully treated for HIV, HCV or both.


A review of acid sulfate soil impacts, actions and policies that impact on water quality in Great Barrier Reef catchments, including a case study on remediation at East Trinity.
An estimated 666,000 ha of acid sulfate soils (ASS) occur within the Great Barrier Reef (GBR) catchments of Queensland, Australia. Extensive areas have been drained causing acidification, metal contamination, deoxygenation and iron precipitation in reef receiving waters. The close proximity of ASS to reef waters makes them a substantial threat to water quality. Another important issue linked with ASS is their release of soluble iron, which is known to stimulate nuisance marine algal blooms, in particular Lyngbya majuscula. Known blooms of the cyanobacteria in reef waters have been confirmed at Shoalwater Bay, Corio Bay, the Whitsunday area and Hinchinbrook Channel. Acid sulfate soils are intimately related to coastal wetland landscapes. Where landscapes containing ASS have been disturbed (such as for agriculture, aquaculture, marinas, etc.) the biodiversity of adjacent wetlands can be adversely affected. However, there is no clear knowledge of the real extent of the so-called "hotspot" ASS areas that occur within the GBR catchments. Management of ASS in reef catchments has benefited from the implementation of the Queensland Acid Sulfate Soils Management Strategy through policy development, mapping, training programs, an advisory service, research and community participation. However, major gaps remain in mapping the extent and nature of ASS. Areas of significant acidification (i.e. hotspots) need to be identified and policies developed for their remediation. Research has a critical role to play in understanding ASS risk and finding solutions, to prevent the adverse impacts that may be caused by ASS disturbance. A case study is presented of the East Trinity site near Cairns, a failed sugar cane development that episodically discharges large amounts of acid into Trinity Inlet, resulting in periodic fish kills. Details are presented of scientific investigations, and a lime-assisted tidal exchange strategy that are being undertaken to remediate a serious ASS problem.


Phylogeny of related functions: the case of polyamine biosynthetic enzymes.
Genome annotation requires explicit identification of gene function. This task frequently uses protein sequence alignments with examples having a known function. Genetic drift, co-evolution of subunits in protein complexes and a variety of other constraints interfere with the relevance of alignments. Using a specific class of proteins, it is shown that a simple data analysis approach can help solve some of the problems posed. The origin of ureohydrolases has been explored by comparing sequence similarity trees, maximizing amino acid alignment conservation. The trees separate agmatinases from arginases but suggest the presence of unknown biases responsible for unexpected positions of some enzymes. Using factorial correspondence analysis, a distance tree between sequences was established, comparing regions with gaps in the alignments. The gap tree gives a consistent picture of functional kinship, perhaps reflecting some aspects of phylogeny, with a clear domain of enzymes encoding two types of ureohydrolases (agmatinases and arginases) and activities related to, but different from ureohydrolases. Several annotated genes appeared to correspond to a wrong assignment if the trees were significant. They were cloned and their products expressed and identified biochemically. This substantiated the validity of the gap tree. Its organization suggests a very ancient origin of ureohydrolases. Some enzymes of eukaryotic origin are spread throughout the arginase part of the trees: they might have been derived from the genes found in the early symbiotic bacteria that became the organelles. They were transferred to the nucleus when symbiotic genes had to escape Muller's ratchet. This work also shows that arginases and agmatinases share the same two manganese-ion-binding sites and exhibit only subtle differences that can be accounted for knowing the three-dimensional structure of arginases. In the absence of explicit biochemical data, extreme caution is needed when annotating genes having similarities to ureohydrolases.


Non-operative management of humeral periprosthetic fracture after stemless shoulder arthroplasty.
Periprosthetic fractures around a stemless implant often involve lesser and greater tuberosities with a well-fixed implant in the metaphysis. This exposes the surgeon to unique questions and challenges as no surgical option (open reduction and internal fixation or revision to a stem) appears satisfactory to address them. Purpose of this study was to evaluate the clinical outcomes after non-operative management of periprosthetic fractures after stemless shoulder arthroplasty. A retrospective multicenter study was conducted to identify all patients who had sustained non-operative management of a periprosthetic fracture after a stemless shoulder. Exclusion criteria were as follows: (1) intraoperative fractures and (2) implant loosening. Primary outcomes included mean Constant score and mean active range of motion. Secondary outcomes were VAS, radiological analysis, and complications. Nine patients were included. One was excluded due to the loss of follow-up at three months. Mean age was 79 years. At the last follow-up, no significant difference was observed between the Constant score, VAS, or the range of motion before fracture and at the last follow-up. Fracture healing did not result in any change in angulation in the frontal plane in seven cases and was responsible for a varus malunion in two cases of anatomic arthroplasty. No change in lateralization or distalization was reported. No cases of implant loosening after fracture have been observed. Conservative management seems to be appropriate in cases of minimally displaced fractures without implant loosening.


Impact of cervical screening on cervical cancer mortality: estimation using stage-specific results from a nested case-control study.
It is well established that screening can prevent cervical cancer, but the magnitude of the impact of regular screening on cervical cancer mortality is unknown. Population-based case-control study using prospectively recorded cervical screening data, England 1988-2013. Case women had cervical cancer diagnosed during April 2007-March 2013 aged 25-79 years (N=11 619). Two cancer-free controls were individually age matched to each case. We used conditional logistic regression to estimate the odds ratio (OR) of developing stage-specific cancer for women regularly screened or irregularly screened compared with women not screened in the preceding 15 years. Mortality was estimated from excess deaths within 5 years of diagnosis using stage-specific 5-year relative survival from England with adjustment for age within stage based on SEER (Surveillance, Epidemiology and End Results, USA) data. In women aged 35-64 years, regular screening is associated with a 67% (95% confidence interval (CI): 62-73%) reduction in stage 1A cancer and a 95% (95% CI: 94-97%) reduction in stage 3 or worse cervical cancer: the estimated OR comparing regular (⩽5.5yearly) screening to no (or minimal) screening are 0.18 (95% CI: 0.16-0.19) for cancer incidence and 0.08 (95% CI: 0.07-0.09) for mortality. It is estimated that in England screening currently prevents 70% (95% CI: 66-73%) of cervical cancer deaths (all ages); however, if everyone attended screening regularly, 83% (95% CI: 82-84%) could be prevented. The association between cervical cancer screening and incidence is stronger in more advanced stage cancers, and screening is more effective at preventing death from cancer than preventing cancer itself.


Identification of a serpin specifically expressed in multipotent and bipotent hematopoietic progenitor cells and in activated T cells.
We have identified a gene that has a high level of mRNA expression in undifferentiated, multipotential hematopoietic cells (FDCP-Mix) and that downregulates both transcript and protein, as these cells are induced to differentiate into mature myeloid cells. Sequence analysis of this gene has identified it as a serine protease inhibitor EB22/3 (serpin 2A). Constitutive expression of serpin 2A in FDCP-Mix cells was associated with an increase in the clonogenic potential of the cells and with a delay in the appearance of fully mature cells in cultures undergoing granulocyte macrophage differentiation when compared with control cells. Serpin 2A was also found to be expressed in bone marrow-derived bipotent granulocyte macrophage progenitor cells (GM-colony forming cell [CFC]), but not in erythrocyte progenitor cells from day 15 fetal liver. Expression of serpin 2A also showed a marked up regulation during the activation of cytotoxic suppressor CD8+ T cells, with a clear lag between the appearance of transcript and detection of protein.


[A master's exam in surgical training].
AN EXAMPLE FOR THE PRESENT DAY: The current requirement for explicit quality standards and examination of surgeons is an opportunity to contemplate surgical training from a historical perspective by looking at the regulations of the Amsterdam Surgeons' Guild (1461-1736). At that time Amsterdam surgeons usually trained for five years in a master-apprentice relationship under the guidance of a master surgeon in a surgeon's shop. An important part of the surgical training took place in the botanical gardens and anatomical theatre, where, during the weekly lessons, the praelector anatomiae would also demonstrate anatomy on the bodies of the deceased. Surgical training was complete after the trainee had passed the 'meesterproef' (master's exam), in which the manufacturing of lancets, blood-letting and performing a trepanation on a skull played a major part. However, over the course of time the final master's exam as the ultimate test of capability at the end of surgical training has disappeared. From the perspective of renewed interest in explicit quality standards and examination of surgeons, the reintroduction of a modern master's exam should perhaps be considered.


Glutamate and GABA-mediated synaptic currents in neurons of the rat dorsal motor nucleus of the vagus.
We report the presence of excitatory and inhibitory spontaneous and evoked synaptic currents in the dorsal motor nucleus of the vagus (DMV) in the rat upon vagal and perivagal stimulation. Whole cell current-clamp recordings from anatomically identified DMV neurons in rat brain stem slices show that these neurons are capable of sustained slow-frequency action potential firing probably because of the presence of pacemaker current. Spontaneously occurring, tetrodotoxin-resistant miniature inhibitory and excitatory synaptic potentials were observed. Stimulation of the vagus mostly induced antidromic action potentials in DMV neurons. However, careful positioning of the stimulating electrode in the tissue surrounding the recording neuron, and sometimes in the vagus itself, was capable of evoking orthodromic-evoked mixed inhibitory-excitatory postsynaptic potentials, and eventually, action potentials. Whole cell voltage-clamp recordings of the synaptic currents corresponding to these synaptic potentials in the presence of pharmacological antagonists of the neurotransmitters gamma-aminobutyric acid (GABA), glutamate, and glycine receptor subtypes indicate that the inhibitory synaptic currents are mediated by GABA-activated Cl- channels, while the excitatory synaptic currents are due to activation of ionotropic glutamate receptors of the N-methyl-D-aspartic acid (NMDA) and non-NMDA subtypes.


Perspectives in musculoskeletal injury management by traditional bone setters in Ashanti, Ghana.
The popularity of the services of traditional bone setters (TBS) in Ghana as an alternative health care requires exploration and documentation of the perspectives of providers and users. To explore and document the perspectives of providers and users of the services of TBS in the management of musculoskeletal injuries in the Ashanti region, Ghana. From the social constructivist and qualitative approach, in-depth interviews were used to explore the perspectives of eight TBS and 16 users of their services, selected purposively through snowballing. Thematic content analysis (TCA) was employed. High recovery rate, warm reception, prompt attention, and the relatively lower charges, are reported to motivate the patronage of the services of TBS for the management of fractures in the legs, arms, ribs, joint bones dislocations, waist and spinal cord problems. The TBS combined traditional and orthodox procedures, using plant and animal-based materials, beliefs, spirituality (God-given) and physical therapy in the management of musculoskeletal injuries. No adverse experience was reported by either the providers or users of the traditional management methods. With plant and animal-based materials, TBS are observed to combine traditional and orthodox procedures to confidently manage musculoskeletal injuries to the satisfaction of their highly motivated patrons. Although over 60% of the TBS attribute the healing power behind their practice to God, the rest do not discount the role of spiritual therapy. Further studies expanded to include the perspectives of non-users of the services of the TBS will authenticate the findings of this study.


Family screening in hypertrophic cardiomyopathy is underperformed, but can be improved by a specialised clinic.
Hypertrophic cardiomyopathy (HCM) causes significant morbidity and sudden death. First-degree relatives (FDR) of affected patients are at risk due to autosomal dominant inheritance. Guidelines recommend clinical screening, including echocardiography, for all FDR. We sought to determine adherence to these guidelines, and whether a specialised HCM clinic improves screening rates. This 12-month prospective follow-up study obtained family pedigrees from all patients referred to the HCM Clinic @ The Alfred. The number of living FDR was determined, and whether they had previously been assessed by echocardiography. One year after a coordinated clinic-based family screening approach was instituted, the number of additionally screened FDR was recorded. Three hundred and eight living FDR of 61 HCM patients were identified. Of these, echocardiography had previously been performed in only 80 (26%), yielding 13 (16%) additional cases of HCM. Twelve months after attendance at our clinic, 51 additional FDR were screened (64% improvement) and 8 new cases of HCM were identified. Recommended family screening for HCM is underperformed, resulting in missed opportunities to detect subclinical HCM. A coordinated approach through a specialised HCM clinic improves screening rates, thus referral to such a service should be considered for all patients with HCM and their families.


Comparison of the Effects of the 1975 Japanese Diet and the Modern Mediterranean Diet on Lipid Metabolism in Mice.
The Japanese diet and the Mediterranean diet are both known to be good for health, but there had been no direct comparison of their health benefits. In this study, we compared the 1975 Japanese diet, which has been found to have high health benefits, with the 2010 Italian diet, which contributes to the longest life expectancy in Mediterranean countries. Diets were created using one-week menus of the two diets based on FAOSTAT Food Balance Sheets. The diets were prepared, freeze-dried, powdered and fed to mice for 4 weeks to examine their effects on lipid metabolism. In mice fed the Japanese diet, the visceral fat weight was lower, adipocytes were smaller, the liver weight was lower and liver TG tended to be lower than those fed the Italian diet, and little lipid accumulation was observed in hepatocytes of mice fed the Japanese diet. In addition, in mice fed the Japanese diet, the expression levels of genes related to fatty acid synthesis were lower, whereas those of genes related to catabolism of fatty acids and cholesterol were higher than those fed the Italian diet. Therefore, the Japanese diet reduced accumulation of lipids in the white adipose tissue and liver by suppressing fatty acid synthesis and promoting catabolism of fatty acids and cholesterol in the liver, compared to the Italian diet.


Pre-ovulatory LH/hCG surge decreases C-type natriuretic peptide secretion by ovarian granulosa cells to promote meiotic resumption of pre-ovulatory oocytes.
In mammalian follicles, oocytes are arrested at the diplotene stage of prophase I until meiotic resumption following the LH surge. Recently, C-type natriuretic peptide (CNP), encoded by natriuretic peptide precursor type C (NPPC) was found to suppress mouse oocyte maturation by promoting cyclic guanosine 5'-monophospate (cGMP) production in cumulus cells. However, regulation of NPPC/CNP expression during the pre-ovulatory period and their regulation by the LH surge have not been investigated. Based on genome-wide analysis of DNA microarray data sets using samples from periovulatory ovaries, we found increases in NPPC transcripts in granulosa cells during pre-ovulatory follicle growth in mice and a rapid decline induced by the pre-ovulatory LH/hCG stimulation. Treatment of pre-ovulatory animals with hCG decreased ovarian CNP content. In isolated ovarian cells, NPPC mRNA was predominantly expressed in mural granulosa cells exhibiting similar regulation following gonadotrophin treatment. In cultured mouse pre-ovulatory follicles, meiosis resumption in oocytes by hCG treatment was accompanied by decreases in NPPC transcript levels. In cultured mouse cumulus cell-oocyte complexes, CNP treatment inhibited the resumption of meiosis with increases in cGMP levels in both cumulus cells and oocytes. In human ovaries, CNP levels in ovarian follicular fluid were also decreased following treatment of patients with an ovulatory dose of hCG. Our findings demonstrate gonadotrophins regulation of NPPC/CNP expression in mouse and human ovaries and confirm the role of CNP as a potent paracrine oocyte maturation inhibitor.


Towards combinatorial targeted therapy in melanoma: from pre-clinical evidence to clinical application (review).
Over the last few years, clinical trials with BRAF and mitogen-activated protein/extracellular signal-regulated kinase (MEK) inhibitors have shown significant clinical activity in melanoma, but only a fraction of patients respond to these therapies, and development of resistance is frequent. This has prompted a large set of preclinical studies looking at several new combinatorial approaches of pathway- or target-specific inhibitors. At least five main drug association strategies have been verified in vitro and in preclinical models. The most promising include: i) vertical targeting of either MEK or phosphoinositide-3 kinase (PI3K)/mammalian target of rapamycin (mTOR) pathways, or their combined blockade; ii) association of receptor tyrosine kinases (RTKs) inhibitors with other pro-apoptotic strategies; iii) engagement of death receptors in combination with MEK-, mTOR/PI3K-, histone deacetylase (HDAC)-inhibitors, or with anti-apoptotic molecules modulators; iv) strategies aimed at blocking anti-apoptotic proteins belonging to B-cell lymphoma (Bcl-2) or inhibitors of apoptosis (IAP) families associated with MEK/BRAF/p38 inhibition; v) co-inhibition of other molecules important for survival [proteasome, HDAC and Signal transducers and activators of transcription (Stat)3] and the major pathways activated in melanoma; vi) simultaneous targeting of multiple anti-apoptotic molecules. Here we review the anti-melanoma efficacy and mechanism of action of the above-mentioned combinatorial strategies, together with the potential clinical application of the most promising studies that may eventually lead to therapeutic benefit.


Prospective observational measurement of tracheal tube cuff pressures in the emergency department.
Tracheal mucosal blood flow is impaired when tracheal tube cuff pressure is above 30 cm of water, with the potential for tracheal mucosal necrosis. Previous studies have found excessive cuff pressures in simulated patients intubated by North American emergency physicians as well as patients intubated in the prehospital setting and emergency department (ED). This study assessed whether patients intubated in a UK prehospital setting or ED had excessive cuff pressures. Prospective observational study in five ED in southwest England over a 2-month period. All patients over 18 years and intubated in the prehospital setting or in the ED were included. Clinical staff independent of the patients' care recorded the following: age, sex, presenting complaint and indication for intubation, tube size and cuff pressure. Neither the paramedics nor the participating ED staff were aware of the study purpose. Cuff pressure measurements were recorded using a standardised cuff inflator pressure gauge. 61 patients were recruited. The median and mean cuff pressures were 58 and 62 cm of water, respectively. 75% of patients had a cuff pressure greater than 30 cm of water. The median cuff pressures in those patients intubated by senior emergency physicians, junior emergency physicians and paramedics were 70, 46 and 79 cm of water, respectively. Excessive tracheal tube cuff pressures were demonstrated in the majority of patients intubated both in the prehospital setting and ED. This is in keeping with existing evidence. Early measurement and adjustment of cuff pressures is recommended for those patients who require ongoing care.


Benefits of Expressive Writing on Healthcare Workers' Psychological Adjustment During the COVID-19 Pandemic.
COVID-19 outbroke in Wuhan, China, in December 2019 and promptly became a pandemic worldwide, endangering health and life but also causing mild-to-severe psychological distress to lots of people, including healthcare workers (HCWs). Several studies have already showed a high prevalence of depression, anxiety, and post-traumatic symptoms in HCWs but less is known about the efficacy of psychological interventions for relieving their mental distress. The aims of this study were: (1) to evaluate the psychological adjustment of Italian HCWs during the COVID-19 pandemic; (2) to investigate the efficacy of an expressive writing (EW) intervention, based on Pennebaker's paradigmatic protocol, on their psychological adjustment; (3) to analyze if outcomes of EW vary in function of individual differences (age, gender, marital status, and baseline values of symptoms). Fifty-five HCWs were randomly assigned to one of two writing conditions: EW (n = 30) or neutral writing (NW; n = 25). Psychological adjustment (in terms of ptsd, depression and global psychopathology's symptoms, perceived social support, and resilience) was assessed before and after three writing sessions. Participants who received the EW intervention showed higher improvements in ptsd, depression, and global psychopathology symptoms. Improvements in EW group varied in function of age, gender, marital status, and baseline values: young, men, married participants and those who had higher baseline scores showed a higher reduction of psychological distress symptoms while women, single and those who had lower baseline value showed increased social support, and resilience. In conclusion, the EW intervention had positive effects which varied in function of individual differences on HCWs' psychological health.


[Prevalence of mental diseases in women of an urban area].
The main aim was to calculate the prevalence of mental pathology in women between 18 and 70 in a Health District of Pamplona; second, to describe comorbidity and to analyse how mental pathology was recorded in the clinical histories. An observational crossover study with randomised selection. A community study in the Txantrea quarter of Pamplona, covering 21,590 inhabitants, with 7605 women between 18 and 70. Randomised sample, stratified by age, of 237 women between 18 and 70 taken from the 1991 Census. In a face-to-face interview at the Health Centre, the DIS Questionnaire, which diagnoses mental illness, was administered to all participants. A check was made to see if mental pathology was recorded in their clinical history. The prevalence of mental illnesses, mainly Phobias and Depression, in the "last year of life" was 33.3% (27.5-39.5), which fell to 24.9% (19.7-30.7) when tobacco abuse was excluded. The most common pathologies were: Depression (17.3%), Tobacco dependency (17.3%), simple Phobia (14.8%), Agoraphobia (13.5%), social Phobia (8.9%) and post-traumatic stress (8.0%). Understanding the high psychological morbidity in these urban women can contribute to the development of Mental Health Promotion and Prevention Programmes and foment fuller mental health training for Primary Care professionals.


Gene therapy for late infantile neuronal ceroid lipofuscinosis: neurosurgical considerations.
The authors conducted a phase I study of late infantile neuronal ceroid lipofuscinosis using an adenoassociated virus serotype 2 (AAV2) vector containing the deficient CLN2 gene (AAV2(CU)hCLN2). The operative technique, radiographic changes, and surgical complications are presented. Ten patients with late infantile neuronal ceroid lipofuscinosis disease each underwent infusion of AAV2(CU)hCLN2 (3 x 10(12) particle units) into 12 distinct cerebral locations (2 depths/bur hole, 75 minutes/infusion, and 2 microl/minute). Innovative surgical techniques were developed to overcome several obstacles for which little or no established techniques were available. Successful infusion relied on preoperative stereotactic planning to optimize a parenchymal target and diffuse administration. Six entry sites, each having 2 depths of injections, were used to reduce operative time and enhance distribution. A low-profile rigid fixation system with 6 integrated holding arms was utilized to perform simultaneous infusions within a practical time frame. Dural sealant with generous irrigation was used to avoid CSF egress with possible subdural hemorrhage or altered stereotactic registration. Radiographically demonstrated changes were seen in 39 (65%) of 60 injection sites, confirming localization and infusion. There were no radiographically or clinically defined complications. The neurosurgical considerations and results of this study are presented to offer guidance and a basis for the design of future gene therapy or other clinical trials in children that utilize direct therapeutic delivery.


Changes in abundance or structure of the per gene product can alter periodicity of the Drosophila clock.
The period (per) locus, which controls biological rhythms in Drosophila, was originally defined by three chemically induced mutations. Flies carrying the pero mutation were arrhythmic, whereas pers and perl mutants had circadian behavioural rhythms with 19-hour and 29-hour periodicities, respectively. Wild-type flies have 24-hour rhythms. Here we compare the per locus DNA sequences of the three mutants with the parental wild-type. The pers and perl mutations lead to amino-acid substitutions, whereas pero introduces an early translation stop (amber). The results indicate that the protein product of per controls biological rhythms. We also report that the abundance of this protein may set the pace of the Drosophila clock. Although circadian rhythms are restored when arrhythmic (per-) Drosophila are transformed with per locus DNA, flies receiving identical transforming DNA segments can produce rhythms with periods that differ by more than 12 hours. Transcription studies reveal a tenfold variation in the level of per RNA among transformed lines. Levels of per RNA are inversely correlated with period length, so that flies with lowest levels of the per product have slow-running biological clocks. On the basis of the combined studies we suggest that perl and pers mutants produce hypoactive and hyperactive per proteins, respectively.