Developing telepsychiatry services in KwaZulu-Natal -- an action research study.
In 2009, the Departments of Psychiatry and Telehealth of the University of KwaZulu-Natal (UKZN) initiated a three year project to develop telepsychiatry services in KwaZulu-Natal. This paper describes the challenges and opportunities of this project. This was a collaborative, in situ health service project and an action research framework was adopted. Over the three years, following a needs assessment and an e-health Readiness Assessment, two action research cycles were completed. Cycle 1: The preparation, implementation and evaluation of educational material, technical requirements and clinical guidelines and clinical support materials. Cycle 2: Preparation and evaluation of educational and clinical outreach sessions. Videoconference education sessions were beneficial to health staff without post-qualification psychiatry training. A flexible format for education improved knowledge outcomes. Clinical sessions are feasible, but require administration and technical support for facilitation. With facilitator training, 128Kbps is a suitable bandwidth for education but 384Kbps is preferred for clinical consultation. Telepsychiatry offers tremendous potential to facilitate the three strategies suggested by the World Psychiatric Association to address the treatment gap, save time and costs and improve access to the small pool of specialist psychiatrists in resource constrained environments.


Non-invasive in vivo monitoring of trackable viruses expressing soluble marker peptides.
Noninvasive methods are needed to study the kinetic properties of viruses in living organisms. Oncolytic viruses are used increasingly for cancer therapy but there is currently no satisfactory way to measure efficiency of tumor transduction, changing levels of viral gene expression or the timing of virus elimination. We therefore generated trackable oncolytic measles viruses expressing inert (nonimmunogenic, nonfunctional and accurately measurable) soluble marker peptides. The marker peptides did not compromise virus replication. Ex vivo and in vivo kinetics of the trackable viruses could be easily followed by measuring the concentrations of virally encoded marker peptides in culture supernatant or in serum. When mice bearing human tumor xenografts were challenged with the trackable viruses, distinct kinetic profiles of marker-gene expression could be correlated with distinct therapeutic outcomes. Oncolytic viruses expressing inert soluble marker polypeptides should greatly facilitate the rational development of effective, individually tailored cancer virotherapy.


Outcome of cardiopulmonary resuscitation in patients on chronic dialysis.
The authors analyzed the outcome of 56 episodes of cardiopulmonary resuscitation (CPR) in dialysis patients. Eleven patients (20%) left the hospital alive. Univariate analysis showed that a functional cause of cardiac arrest, absence of rib fractures, and the occurrence of cardiac arrest in the dialysis or intensive care units were associated with a favorable outcome. Logistic regression showed that the outcome of CPR was related to the presence of rib fractures, cause of arrest, degree of preexisting heart disease, and patient age. Despite the high incidence of rib fractures (77%), the outcome of CPR in dialysis patients is similar to its outcome in the general population.


Sodium, potassium, body mass, alcohol and blood pressure in three United Kingdom centres (the INTERSALT study).
Standardised data on blood pressure, 24 h urinary electrolyte excretion, body mass index (BMI) and alcohol intake were collected as part of the INTERSALT study in 598 men and women aged 20-59 years, selected randomly from three population groups in the United Kingdom. For the three centres combined, mean systolic blood pressure was 121.4 mm Hg and diastolic pressure 72.1 mm Hg, urinary sodium excretion 152.1 mmol/24 h, urinary potassium excretion 61.0 mmol/24 h, urinary sodium/potassium ratio 2.64 and BMI 25.2 kg/m2. Prevalence of heavy alcohol drinking in men (greater than or equal to 300 ml/week) was 27.5 per cent. Applying overall INTERSALT regression coefficients to the United Kingdom data suggested that modest changes in average sodium and potassium intakes, together with reductions in the prevalence of obesity and (in men) of heavy alcohol drinking could lead to important reductions in average population blood pressures and the prevalence of hypertension. The potential of this multifactorial approach to blood pressure control was illustrated by stratifying individuals within each of the United Kingdom centres by sodium and potassium excretion, BMI and alcohol intake. The 20 (out of 299) men considered at 'lower risk' for high blood pressure with respect to the above variables had systolic pressure lower by 11 mm Hg (P less than 0.01); for the 27 (out of 299) 'lower risk' women, systolic pressure was lower by 5 mm Hg (P = 0.06). These non-pharmacological approaches towards more favourable blood pressure levels could be accompanied by reductions in mortality from stroke and coronary heart disease.


Financial incentives to discontinue long-term benzodiazepine use: a discrete choice experiment investigating patient preferences and willingness to participate.
Investigate the acceptability of financial incentives for initiating a medically supervised benzodiazepine discontinuation programme among people with long-term benzodiazepine use and to identify programme features that influence willingness to participate. We conducted a discrete choice experiment in which we presented a variety of incentive-based programs to a sample of older adults with long-term benzodiazepine use identified using the outpatient electronic health record of a university-owned health system. We studied four programme variables: incentive amount for initiating the programme, incentive amount for successful benzodiazepine discontinuation, lottery versus certain payment and whether partial payment was given for dose reduction. Respondents reported their willingness to participate in the programmes and additional information was collected on demographics, history of use and anxiety symptoms. The overall response rate was 28.4%. Among the 126 respondents, all four programme variables influenced stated preferences. Respondents strongly preferred guaranteed cash-based incentives as opposed to a lottery, and the dollar amount of both the starting and conditional incentives had a substantial impact on choice. Willingness to participate increased with the amount of conditional incentive. Programme participation also varied by gender, duration of use and income. Participation in an incentive-based benzodiazepine discontinuation programme might be relatively low, but is modifiable by programme variables including incentive amounts. These results will be helpful to inform the design of future trials of benzodiazepine discontinuation programmes. Further research is needed to assess the financial viability and potential cost-effectiveness of such economic incentives.


Molecular phylogenetic position and the life cycle of Phyllodistomum cyprini Feng et Wang, 1995 in Japan, with a note of a larval Phyllodistomum sp. from unionid mussels in North America.
A gorgoderid species, Phyllodistomum cyprini Feng et Wang, 1995 obtained from ureters of common carp, Cyprinus carpio in Japan is described both morphologically and molecularly. Its larval stage is a macrocercous cercaria found in a unionid mussel, Nodularia nipponensis first described by Urabe et al. (2015). A molecular phylogenetic study revealed that P. cyprini is not closely related to rhopalocercous cercariae from unionid mussels both in Europe and North America. This result indicates that there are several distinct clades in Phyllodistomum species that use unionid mussels as a first intermediate host, and suggests that the cercarial morphology may be a more accurate indicator of the phylogeny of Phyllodistomum than molluscan host identity.


A Content Analysis of Teen Parenthood in "Teen Mom" Reality Programming.
A content analysis of the MTV shows 16 and Pregnant (n = 59), Teen Mom (n = 20), and Teen Mom 2 (n = 20) was conducted to determine whether these programs accurately portray teen pregnancy. The results revealed that teen mothers on 16 and Pregnant were younger, more often White, and had more healthy babies as compared to national averages. The babies' fathers were more involved in the daily care of their child as compared to reality. Medical insurance or receipt of government assistance was almost never discussed. Teen mothers in the Teen Mom shows were significantly more likely to achieve a high school diploma as compared to reality. Finally, mothers on Teen Mom and Teen Mom 2 were significantly less likely to voice concern about finances and had more active social lives than mothers on 16 and Pregnant. Using social learning theory as a theoretical framework, we argue that these shows provide mixed messages to young audiences about teen pregnancy and parenthood.


Polypurine/polypyrimidine sequences as cis-acting transcriptional regulators.
Genome sequence information has generated increasing evidence for the claim that repetitive DNA sequences present within and around genes could play a important role in the regulation of gene expression. Polypurine/polypyrimidine sequences [poly(Pu/Py)] have been observed in the vicinity of promoters and within the transcribed regions of many genes. To understand whether such sequences influence the level of gene expression, we constructed several prokaryotic and eukaryotic expression vectors incorporating poly(Pu/Py) repeats both within and upstream of a reporter gene, lacZ (encoding beta-galactosidase), and studied its expression in vivo. We find that, in contrast to the situation in Escherichia coli, the presence of poly(Pu/Py) sequences within the gene does not significantly inhibit gene expression in mammalian cells. On the other hand, the presence of such sequences upstream of lacZ leads to a several-fold reduction of gene expression in mammalian cells. Similar down-regulation was observed when a structural cassette containing poly(Pu/Py) sequences upstream of lacZ was integrated into yeast chromosome V. Sequence analysis of the nine totally sequenced yeast chromosomes shows that a large number of such sequences occur upstream of ORFs. On the basis of our experimental results and DNA sequence analysis, we propose that these sequences can function as cis-acting transcriptional regulators.


Nipah virus infection: current scenario.
The emergence of Nipah virus (NiV) infection into the pig population and subsequently into the human population is believed to be due to changes in ecological conditions. In Malaysia, A major NiV outbreak occurred in pigs and humans from September 1998 to April 1999 that resulted in infection of 265 and death of 105 persons. About 1.1 million pigs had to be destroyed to control the outbreak. The disease was recorded in the form of a major outbreak in India in 2001 and then a small incidence in 2007, both the outbreaks in West Bengal only in humans without any involvement of pigs. There were series of human Nipah incidences in Bangladesh from 2001 till 2013 almost every year with mortality exceeding 70 %. The disease transmission from pigs acting as an intermediate host during Malaysian and Singapore outbreaks has changed in NIV outbreaks in India and Bangladesh, transmitting the disease directly from bats to human followed by human to human. The drinking of raw date palm sap contaminated with fruit bat urine or saliva containing NiV is the only known cause of outbreak of the disease in Bangladesh outbreaks. The virus is now known to exist in various fruit bats of Pteropus as well as bats of other genera in a wider belt from Asia to Africa.


Parallel pathways for recruiting effector proteins determine centromere drive and suppression.
Selfish centromere DNA sequences bias their transmission to the egg in female meiosis. Evolutionary theory suggests that centromere proteins evolve to suppress costs of this "centromere drive." In hybrid mouse models with genetically different maternal and paternal centromeres, selfish centromere DNA exploits a kinetochore pathway to recruit microtubule-destabilizing proteins that act as drive effectors. We show that such functional differences are suppressed by a parallel pathway for effector recruitment by heterochromatin, which is similar between centromeres in this system. Disrupting the kinetochore pathway with a divergent allele of CENP-C reduces functional differences between centromeres, whereas disrupting heterochromatin by CENP-B deletion amplifies the differences. Molecular evolution analyses using Murinae genomes identify adaptive evolution in proteins in both pathways. We propose that centromere proteins have recurrently evolved to minimize the kinetochore pathway, which is exploited by selfish DNA, relative to the heterochromatin pathway that equalizes centromeres, while maintaining essential functions.


Twenty-eight cases of human heart-lung transplantation.
Between March, 1981, and August, 1985, twenty-eight heart-lung transplant operations were done in 27 patients at a single institution. 8 patients died in the perioperative period and adhesions related to previous thoracic surgery proved to be a major risk factor for postoperative haemorrhage. Obliterative bronchiolitis developed in half of the 20 long-term survivors, a mean of 11.2 months (range 2-35 months) after surgery: 4 of these patients died, 3 are functionally limited, 2 were successfully treated with corticosteroids, and the remaining patient was successfully retransplanted. The other 10 long-term survivors returned to a normal life with essentially normal pulmonary function measured at a mean of 22.6 months (range 4-42 months) after transplantation. All the surviving patients have evidence of renal impairment related to cyclosporin nephrotoxicity. The results indicate that, although heart-lung transplantation is compatible with essentially normal long-term pulmonary function, the procedure should not yet be regarded as a routine clinical intervention.


Lights, camera, active! appreciation of active learning predicts positive attitudes towards lecture capture.
Much has been written about instructor attitudes towards lecture capture, particularly concerning political issues such as opt-out policies and the use of recordings by management. Additionally, the pedagogical concerns of lecturers have been extensively described and focus on the belief that recording lectures will impact on attendance and will reduce interactivity and active learning activities in lectures. However, little work has looked at the relationship between attitudes towards lecture capture and broader conceptions of learning and teaching. In this pre-registered study, we administered the Conceptions of Learning and Teaching scale and a novel lecture capture attitude scale to 159 higher education teachers. We found that appreciation of active learning predicted more positive attitudes towards lecture recordings as an educational support tool, whilst higher teacher-centred scores predicted greater concern about the negative educational impact of recordings. The effects observed were small; however, they are strong evidence against the view that it is instructors who value participatory and active learning that are opposed to lecture capture. Exploratory analyses also suggested that those who did not view recordings as an essential educational resource record fewer of their lectures, highlighting the real-world impact that attitudes can have, and further strengthening the need for staff to be provided with evidence-based guidance upon which to base their teaching practice. Data, analysis code, and the pre-registration are available athttps://osf.io/uzs3t/.


Growth hormone for in vitro fertilization.
Ovulation induction protocols for in vitro fertilization (IVF) are constantly under review and revision in an attempt to decrease gonadotropin requirements while improving follicular recruitment and pregnancy rate. Most studies of the effect of Growth Hormone on ovulation induction have investigated normally ovulating infertile women. Attention has begun to turn to its effect on women who fail to produce an optimal number of follicles in response to ovulation induction. To assess the effectiveness of growth hormone adjuvant therapy for women undergoing ovulation induction prior to IVF in two patient groups: a) those with no previous history of poor response and b) those with a history of poor response. This review has drawn on the search strategy developed for the Subfertility Group as a whole. Relevant trials were identified in the Group's Specialised Register of Controlled Trials. See Review Group details for more information. All RCTs were included if they addressed the research question posed and provided outcome data for intervention and control subjects. Outcomes of interest included pregnancy, oocyte/embryo number and peak estradiol level. A diverse search strategy was employed, including hand-search of 43 core journals from 1966 to the present, bibliographies of relevant trials, MEDLINE database, abstracts from North American and European meetings and contact with authors of relevant papers. Relevant data were extracted independently by two reviewers using the standardized data extraction sheet. Validity was assessed in terms of method of randomization, completeness of follow-up, presence or absence of crossover and co-intervention. 2x2 tables were generated for all relevant outcomes. Odds ratios were generated using the Peto modified Mantel-Haenszel technique. Statistical heterogeneity was assessed using x2. In women with no previous history of poor stimulation, the common odds ratio for pregnancy per cycle was 0.97 (95% CI 0.34-2. 76) Total dose of gonadotropin and number of oocytes retrieved were similar between treated and placebo groups. In previous poor responders, the common odds ratio for pregnancy per cycle instituted was 2.55 (95% CI 0.64-10.12). Again, no statistically significant difference was noted in gonadotropin dosage or oocyte number obtained, when all studies were considered. Data from 6 small trials suggest that in women with no previous poor response, GH augmentation does not improve the rate of pregnancy. In poor responders, a trend towards improved outcome with GH treatment deserves further study.


[Template activity of poly(A+) and poly(A-) messenger RNA of pathogenic arenaviruses].
The total RNA from cells infected with Machupo and Lassa viruses as well as poly(A+) and poly(A-) fractions of the RNA were translated in the cell-free protein synthesizing system from rabbit reticulocytes. The translated products were treated with specific antibodies and analyzed in polyacrylamide gel electrophoresis. Only poly(A-) fraction of RNA coded for the synthesis of NP protein in vitro. The mRNAs for NP protein of Machupo and Lassa viruses are supposed to contain no poly(A) sequences at 3'end, or if they really do, the size of the sequences is not adequate for binding with oligo(dT)-cellulose.


APOE genotype results in differential effects on the peripheral clearance of amyloid-beta42 in APOE knock-in and knock-out mice.
The epsilon4 allele of apolipoprotein E (APOE) is currently the major genetic risk factor identified for Alzheimer's disease (AD). Previous in vivo data from our laboratory has demonstrated that amyloid-beta (Abeta) is rapidly removed from the plasma by the liver and kidney and that the rate of its clearance is affected by ApoE in C57BL/6J and APOE-/- mice. To expand upon these findings, we assessed the peripheral clearance of human synthetic Abeta42 in APOE epsilon2, epsilon3, and epsilon4 knock-in and APOE knock-out mice injected with lipidated recombinant apoE2, E3, and E4 protein. Our results show that APOE does influence the rate at which the mice are able to clear Abeta42 from their bloodstream. Both APOE epsilon4 mice and APOE knock-out mice treated with lipidated recombinant apoE4 demonstrated increased retention of plasma Abeta42 over time compared to APOE epsilon2/APOE knock-out rE2 and APOE epsilon3/APOE knock-out rE3 mice. These findings suggest that the peripheral clearance of Abeta42 is significantly altered by APOE genotype. Given that APOE epsilon4 is a risk factor for AD, then these novel findings provide some insight into the role of ApoE isoforms on the peripheral clearance of Abeta which may impact on clearance from the brain.


[Lice in Mali: frequency of infestation, genotyping, infection rate and case management].
Because lice-transmitted infections are a real public health problem, epidemiological studies in different ecoclimatic zones of Africa are useful. This article aims to describe the frequency of lice infestation, their genotypes, and their infection by pathogens in the regions of Koulikoro and Mopti. A cross-sectional survey allowed us to collect lice from rural populations. Techniques of molecular biology (real-time PCR, standard PCR, and genotyping) were used for analysis of lice samples. Infestation rates were 57% (12/21) among subjects in Diankabou, in the Sahelian zone; 91% (39/43) in Doneguebougou, and 86% (59/69) in Zorocoro, in a savanna zone. The overall lice infestation rate in the samples in the three localities was 83% (110/133). Real-time PCR showed 3% (4/92) of Acinetobacter baumanii but no B. quintana in Diankabou. Phylogenetic analysis of the mitochondrial gene (Cytb) showed that head lice in Mali belong to genotype C. The high frequency of lice infestation in the study population indicates that it would be useful to conduct national epidemiological surveys to estimate the magnitude of this public health problem.


Secondary neurologic injury resulting from nonhypotensive hemorrhage combined with mild traumatic brain injury.
Although the emergency physician often treats patients with multiple injuries, there are relatively few clinically relevant models that mimic these situations. To describe the changes after a hemorrhagic insult superimposed on traumatic brain injury (TBI), anesthetized and ventilated juvenile pigs were assigned to 35% hemorrhage (35H), TBI (via fluid percussion); TBI + 35H, and TBI + 40H (40% hemorrhage). Animals were resuscitated with shed blood and crystalloid. Hemodynamic, metabolic, behavioral, and histologic parameters were assessed for 48 h. In TBI, mean arterial pressure (MAP) was not significantly different from baseline. For TBI + 40H, MAP fell by 60% (p < 0.05). This was corrected with resuscitation. Interestingly, TBI + 35H did not show a fall in MAP, while in 35H, MAP was reduced similarly to the TBI + 40H group. ICP was elevated only initially in the TBI group. In TBI + 40H and TBI + 35H, ICP increased markedly with resuscitation, remaining elevated for 60 min. ICP remained at baseline with 35 H. Hemorrhagic focal cerebal contusions at the gray-white interface were observed in 3/5 of TBI + 40H and 5/7 of TBI + 35H. Despite the presence of subarachnoid hemorrhage (SAH) in all the animals in the TBI alone group, none of these animals demonstrated grossly discernible intraparenchymal injury. There was no evidence of intracranial injury in the 35H group. Only in animals receiving a secondary insult of hemorrhage following the primary TBI were cerebral contusions found. These experiments demonstrate the evolution of cerebral contusions as a form of secondary neurologic injury following resuscitation from traumatic brain injury and hemorrhage, even in the absence of significant blood pressure changes.


Cancer stem cell enrichment marker CD98: a prognostic factor for survival in patients with human papillomavirus-positive oropharyngeal cancer.
Several hypotheses have been proposed to explain the relatively good prognosis of patients with a human papillomavirus (HPV)-positive oropharyngeal squamous cell carcinoma (OPSCC) and one of these is a higher sensitivity to (chemo)radiation. Previous studies have suggested that treatment failure in OPSCC patients is caused by resistance of cancer stem cells (CSCs). The purpose of this study was to evaluate the association between the number of CSCs and prognosis in HPV-positive OPSCC patients. All OPSCC patients (n=711) treated between 2000 and 2006 in two Dutch university hospitals were included. Presence of HPV in a tumour tissue specimen was tested by p16-immunostaining followed by HPV DNA GP5+/6+polymerase chain reaction (PCR). The presence and intensity of tumour CSC markers CD44 and CD98 were determined by immunohistochemistry and semiquantitative scoring was performed. Overall survival (OS) and progression-free survival (PFS) rates were compared between patients with low and high CD44/CD98 expression in relation to HPV status. HPV-positive tumours showed a lower percentage of cells with CD44 and CD98 expression than HPV-negative tumours (p<0.001, χ(2)-test). Within the group of patients with HPV-positive OPSCC, a high percentage of CD98-positive tumour cells was associated with a significantly worse 5-year OS and PFS (OS: 36.4% and PFS: 27.3%) compared to patients with a low percentage of CD98-positive cells (OS: 71.9% and PFS: 70.5%, respectively) (p<0.001). HPV-positive OPSCCs harbour fewer cells expressing the CSC enrichment markers CD44 and CD98. Furthermore, OS and PFS were significantly worse for patients with HPV-positive OPSCC with a high percentage of CD98-positive cells.


Indoor orientation and mobility for learners who are blind.
This study consisted of designing a mobile technology to identify the position and orientation of people who are blind in closed environments, such as a school, building or home, identifying the necessary infrastructure based on criteria of usability, reliability and accessibility, in order for PocketPC device interface developers to be able to define the technology to be used at an early stage. The technology consisted of using PocketPC devices with a data collecting application and another for the representation of space. We evaluated its usability, accessibility, reliability, efficiency, and availability. The results of the usability evaluation assured us that the interface designed and developed is usable for users who are blind, which is reaffirmed through the accessibility evaluation. The reliability, efficiency and availability evaluations of the system show that a user who is blind can use the system confidently, because it provides correct information for more independent navigation.


Positive autoregulation of mrkHI by the cyclic di-GMP-dependent MrkH protein in the biofilm regulatory circuit of Klebsiella pneumoniae.
Klebsiella pneumoniae is an important cause of nosocomial infections, primarily through the formation of surface-associated biofilms to promote microbial colonization on host tissues. Expression of type 3 fimbriae by K. pneumoniae facilitates surface adherence, a process strongly activated by the cyclic di-GMP (c-di-GMP)-dependent transcriptional activator MrkH. In this study, we demonstrated the critical importance of MrkH in facilitating K. pneumoniae attachment on a variety of medically relevant materials and demonstrated the mechanism by which bacteria activate expression of type 3 fimbriae to colonize these materials. Sequence analysis revealed a putative MrkH recognition DNA sequence ("MrkH box"; TATCAA) located in the regulatory region of the mrkHI operon. Mutational analysis, electrophoretic mobility shift assay, and quantitative PCR experiments demonstrated that MrkH binds to the cognate DNA sequence to autoregulate mrkHI expression in a c-di-GMP-dependent manner. A half-turn deletion, but not a full-turn deletion, between the MrkH box and the -35 promoter element rendered MrkH ineffective in activating mrkHI expression, implying that a direct interaction between MrkH and RNA polymerase exists. In vivo analyses showed that residues L260, R265, N268, C269, E273, and I275 in the C-terminal domain of the RNA polymerase α subunit are involved in the positive control of mrkHI expression by MrkH and revealed the regions of MrkH required for DNA binding and transcriptional activation. Taken together, the data suggest a model whereby c-di-GMP-dependent MrkH recruits RNA polymerase to the mrkHI promoter to autoactivate mrkH expression. Increased MrkH production subsequently drives mrkABCDF expression when activated by c-di-GMP, leading to biosynthesis of type 3 fimbriae and biofilm formation. Bacterial biofilms can cause persistent infections that are refractory to antimicrobial treatments. This study investigated how a commonly encountered hospital-acquired pathogen, Klebsiella pneumoniae, controls the expression of MrkH, the principal regulator of type 3 fimbriae and biofilm formation. We discovered a regulatory circuit whereby MrkH acts as a c-di-GMP-dependent transcriptional activator of both the gene cluster of type 3 fimbriae and the mrkHI operon. In this positive-feedback loop, whereby MrkH activates its own production, K. pneumoniae has evolved a mechanism to ensure rapid MrkH production, expression of type 3 fimbriae, and subsequent biofilm formation under favorable conditions. Deciphering the molecular mechanisms of biofilm formation by bacterial pathogens is important for the development of innovative treatment strategies for biofilm infections.


Chemosensory Dysfunctions Induced by COVID-19 Can Persist up to 7 Months: A Study of Over 700 Healthcare Workers.
Several studies have revealed either self-reported chemosensory alterations in large groups or objective quantified chemosensory impairments in smaller populations of patients diagnosed with COVID-19. However, due to the great variability in published results regarding COVID-19-induced chemosensory impairments and their follow-up, prognosis for chemosensory functions in patients with such complaints remains unclear. Our objective is to describe the various chemosensory alterations associated with COVID-19 and their prevalence and evolution after infection. A cross-sectional study of 704 healthcare workers with a RT-PCR-confirmed SARS-CoV-2 infection between 2020 February 28 and 2020 June 14 was conducted 3-7 months after onset of symptoms. Data were collected with an online questionnaire. Outcomes included differences in reported chemosensory self-assessment of olfactory, gustatory, and trigeminal functions across time points and Chemosensory Perception Test scores from an easy-to-use at-home self-administered chemosensory test. Among the 704 participants, 593 (84.2%) were women, the mean (SD) age was 42 (12) years, and the questionnaire was answered on average 4.8 (0.8) months after COVID-19. During COVID-19, a decrease in olfactory, gustatory, and trigeminal sensitivities was reported by 81.3%, 81.5%, and 48.0%, respectively. Three to 7 months later, reduced sensitivity was still reported by 52.0%, 41.9%, and 23.3%, respectively. Chemosensory Perception Test scores indicate that 19.5% of participants had objective olfactory impairment. These data suggest a significant proportion of COVID-19 cases have persistent chemosensory impairments at 3-7 months after their infection, but the majority of those who had completely lost their olfactory, gustatory, and trigeminal sensitivities have improved.


Proinflammatory and vasodilator effects of nociceptin/orphanin FQ in the rat mesenteric microcirculation are mediated by histamine.
Nociceptin/orphanin FQ (N/OFQ) is the endogenous ligand for the N/OFQ peptide receptor (NOP). N/OFQ causes hypotension and vasodilation, and we aimed to determine the role of histamine in inflammatory microvascular responses to N/OFQ. Male Wistar rats (220-300 g, n = 72) were anesthetized with thiopental (30 mg/kg bolus, 40-90 mg x kg(-1) x h(-1) iv), and the mesentery was prepared for fluorescent intravital microscopy using fluorescein isothiocyanate-conjugated BSA (FITC-BSA, 0.25 ml/100 g iv) or 1 microm fluorescently labeled microspheres. N/OFQ (0.6-60 nmol/kg iv) caused hypotension (SAP, baseline: 154 +/- 11 mmHg, 15 nmol/kg N/OFQ: 112 +/- 10 mmHg, P = 0.009), vasodilation (venules: 23.9 +/- 1.2 microm, 26.7 +/- 1.2 microm, P = 0.006), macromolecular leak (interstitial gray level FITC-BSA: 103.7 +/- 3.4, 123.5 +/- 11.8, P = 0.009), and leukocyte adhesion (2.0 +/- 0.9, 15.2 +/- 0.9/100 microm, P = 0.036). Microsphere velocity also decreased (venules: 1,230 +/- 370 microm/s, P = 0.037), but there were no significant changes in blood flow. Flow cytometry measured a concurrent increase in neutrophil expression of cd11b with N/OFQ vs. controls (Geo mean fluorescence: 4.19 +/- 0.13 vs. 2.06 +/- 0.38, P < 0.05). The NOP antagonist [Nphe(1),Arg(14),Lys(15)]N/OFQ-NH(2) (UFP-101; 60 and 150 nmol/kg iv), H(1) and H(2)antagonists pyrilamine (mepyramine, 1 mg/kg iv) and ranitidine (1 mg/kg iv), and mast cell stabilizer cromolyn (1 mg x kg(-1) x min(-1)) also abolished vasodilation and macromolecular leak to N/OFQ in vivo (P < 0.05), but did not affect hypotension. Isolated mesenteric arteries (approximately 200 microm, n = 25) preconstricted with U-46619 were also mounted on a pressure myograph (60 mmHg), and both intraluminally and extraluminally administered N/OFQ (10(-5) M) caused dilation, inhibited by pyrilamine in the extraluminal but not the intraluminal (control: -6.9 +/- 3.8%; N/OFQ: 32.6 +/- 8.4%; pyrilamine: 31.5 +/- 6.8%, n = 18, P < 0.05) experiments. We conclude that, in vivo, mesenteric microvascular dilation and macromolecular leak occur via N/OFQ-NOP-mediated release of histamine from mast cells. Therefore, N/OFQ-NOP has an important role in microvascular inflammation, and this may be targeted during disease, particularly as we have proven that UFP-101 is an effective antagonist of microvascular responses in vivo.


A next-generation-sequencing panel for mutational analysis of dominant acute hepatic porphyrias.
Molecular diagnosis of autosomal dominant acute hepatic porphyrias (AHPs) plays an important role in the management of these disorders. To introduce next generation sequencing (NGS) to the porphyria diagnosis, we designed a panel that contained four genes, ALAS1, HMBS, CPOX and PPOX for mutational analysis of acute intermittent porphyria (AIP), hereditary coproporphyria (HCP) and variegate porphyria (VP). To validate the AHP panel, 30 samples with known pathogenic variants as determined by Sanger sequencing, were analyzed using the Ion PGM™. Among them, nine have so far not been reported. The pathogenic variants were identified and annotated manually in IGV by three individuals who were blinded to the Sanger results. The AHP panel consists of 95 amplicons that covers 92% of the coding region of the four genes. Of the 95 amplicons, 93 had an average read-depth of >500 reads. In 29 of the 30 tested samples, pathogenic variants were correctly identified and annotated. The number of reads from the mutated alleles were approximately 50% of the total. The annotation of a 22-bp duplication with NGS differed from that of Sanger by one nucleotide. NGS showed an advantage in allelic discrimination over Sanger sequencing and was also able to detect a known somatic variant in the HMBS gene. The AHP panel will be applied in the initial diagnosis of new patients. Any sequence variations with a frequency of ≥10% will be confirmed by Sanger sequencing. The cost-effectiveness of a NGS approach for AHP in a diagnostic laboratory needs to be further assessed.


Activation Energy Calculations for Formamide-TiO2 and Formamide-Pt Interactions in the Presence of Water.
Formamide contains the four elements (C, H, O, and N) most required for life and it is attractive as a potential prebiotic starting material for nucleobase synthesis. In the presence of catalysts (for example, TiO2) and with moderate heating, formamide can pass surface energy barriers, yielding a complete set of nucleic bases and acyclonucleosides, and favoring both phosphorylations and transphosphorylations necessary for life. In the reaction mechanism, interaction with water seems to be an essential factor for the formamide molecule to function. In this paper, a formamide-water solution on a TiO$_2$ (anatase) surface is simulated using the molecular dynamics method, and activation energy calculations are performed for the temperature range of T = 250 K to T = 400 K. A correlation is established between the diffusion and density profiles for the formamide and water molecules on an anatase surface. Also, the calculated activation energies of the formamide-water-anatase and formamide-water-platinum systems are compared. A comparative analysis is performed of the behavior of formamide-water and ethanol-water interaction on the same (anatase and platinum) surfaces.


Does Vitamin D Deficiency Contribute to Cognitive Dysfunction in Patients with Systemic Lupus Erythematosus?
Objective: Neurocognitive impairment is one of the most common systemic lupus erythematosus (SLE) manifestations. However, its pathophysiology is still poorly understood. Vitamin D deficiency is a possible risk factor for cognitive impairment. The aim of this study was to evaluate the relationship between 25-dihydroxy(OH) D3 levels and cognitive performance in patients with SLE. Methods: This was a cross-sectional, case-control study that included 30 Egyptian patients diagnosed with SLE and 20 age, sex, and educational level-matched controls. Study participants were subjected to a battery of neuropsychological evaluation using the California Verbal Learning Test (CVLT- II), Controlled Oral Word Association Test (COWAT), and Trail Making Test and evaluation of depression using Beck Depression Inventory (BDI). Serum levels of 25(OH) D3 were measured in the SLE group and control group. Results: The patients with SLE performed worse on total recall of verbal memory and executive function tests than the healthy controls. There was no significant difference between the patients and controls in Beck Depression Inventory (BDI). There was a significant negative correlation between vitamin D levels and executive function assessed by Trail Making Test (r=-0.399, p=0.03). Conclusion: Vitamin D deficiency could have a significant impact on cognitive performance in patients with SLE.


Estimating ideal body mass in children.
An equation and nomogram to estimate ideal body mass (IBM) for establishing dosage regimens of certain drugs in children was derived from a large pediatric data base. The data were from more than 20,000 U.S. children (ages 1-17 years) included in two major surveys published by the National Center for Health Statistics. The IBM was defined as the 50th percentile weight for a given height (ht). The equation derived was IBM(kg) = 2.396e0.01863(ht), where height is in cm. The IBMs estimated from this equation were compared with estimates from three other methods, two of which were derived from an adult population. The adult methods were unsuitable for pediatric use. This equation and nomogram for estimating IBM in children should be useful in determining dosage regimens of aminoglycosides and other drugs for obese pediatric patients.


Cariogenic properties of Streptococcus mutans clinical isolates with sortase defects.
In Streptococcus mutans, a Gram-positive pathogen of dental caries, several surface proteins are anchored by the activity of sortase enzyme. Although various reports have shown that constructed S. mutans mutants deficient of sortase as well as laboratory reference strains with a sortase gene mutation have low cariogenic potential, no known studies have investigated clinical isolates with sortase defects. Here, we examined the cariogenic properties of S. mutans clinical isolates with sortase defects as well as caries status in humans harboring such defective isolates. Sortase-defective clinical isolates were evaluated for biofilm formation, sucrose-dependent adhesion, stress-induced dextran-dependent aggregation, acid production, and acid tolerance. Additionally, caries indices of subjects possessing such defective isolates were determined. Our in vitro results indicated that biofilm with a lower quantity was formed by sortase-defective as compared to non-defective isolates. Moreover, impairments of sucrose-dependent adhesion and stress-induced dextran-dependent aggregation were found among the isolates with defects, whereas no alterations were seen in regard to acid production or tolerance. Furthermore, glucan-binding protein C, a surface protein anchored by sortase activity, was predominantly detected in culture supernatants of all sortase-defective S. mutans isolates. Although the sortase-defective isolates showed lower cariogenic potential because of a reduction in some cariogenic properties, deft/DMFT indices revealed that all subjects harboring those isolates had caries experience. Our findings suggest the impairment of cariogenic properties in S. mutans clinical isolates with sortase defects, though the detection of these defective isolates seemed not to imply low caries risk in the subjects harboring them.


Impact of preadmission variables on USMLE step 1 and step 2 performance.
To examine the predictive ability of preadmission variables on United States Medical Licensing Examinations (USMLE) step 1 and step 2 performance, incorporating the use of a neural network model. Preadmission data were collected on matriculants from 1998 to 2004. Linear regression analysis was first used to identify predictors of performance on step 1 and step 2. A generalized regression neural network (GRNN) as well as a feed forward neural network (FFNN) was then developed in an effort to more accurately predict step 1 and step 2 scores from these preadmission data. Statistically significant predictors for step 1 and step 2 included science grade point average (SGPA), the biologic science (BS) section of the Medical College Admissions Test (MCAT), college selectivity, race, and age of the applicant. Neural networks were found to predict a significant portion of the variance, and the FFNN demonstrated some superiority over that obtained with linear regression models as well as the GRNN. The results have implications that could impact the selection of applicants to medical school and the neural networks that we developed could be used in a prospective manner.


MazEF toxin-antitoxin proteins alter Escherichia coli cell morphology and infrastructure during persister formation and regrowth.
When exposed to antibiotics, many bacteria respond by activating intracellular 'toxin' proteins, which arrest cell growth and induce formation of persister cells that survive antibiotics. After antibiotics are removed, persisters can regrow by synthesizing 'antitoxin' proteins that sequester toxin proteins. In Escherichia coli, MazE antitoxin sequesters the activity of MazF toxin, which extensively cleaves cellular RNAs. Although the functions of MazEF proteins are well characterized, there is surprisingly little known about their effects on cell structure. Here, using a combination of microscopy techniques, we visualized the effects of MazEF and three bactericidal antibiotics on E. coli cell morphology and infrastructure. When ectopically expressed in E. coli, MazF temporarily stalled cell growth and induced persister formation, but only mildly elevated DNA mutagenesis. Viewed by electron microscopy, MazF-expressing persister cells were arrested in cell growth and division. Their chromosomal DNAs were compacted into thread-like structures. Their ribosomes were excluded from their nucleoids. After exposure to ciprofloxacin, persister regrowth was activated by MazE. Cell division remained inhibited while cells became extraordinarily elongated, then divided multiple times during stationary growth phase. This extreme filamentation during persister regrowth was unique to ciprofloxacin-treated persisters, likely caused by inhibition of cell division during regrowth, and was not observed with kanamycin-treated persisters.


Prenatal exposure to persistent organic pollutants and childhood obesity: A systematic review and meta-analysis of human studies.
We conducted a systematic review and meta-analysis of the associations between prenatal exposure to persistent organic pollutants (POPs) and childhood obesity. We focused on organochlorines (dichlorodiphenyltrichloroethane [DDT], dichlorodiphenyldichloroethylene [DDE], hexachlorobenzene [HCB], and polychlorinated biphenyls [PCBs]), perfluoroalkyl and polyfluoroalkyl substances (PFAS), and polybrominated diphenyl ethers (PBDEs) that are the POPs more widely studied in environmental birth cohorts so far. We search two databases (PubMed and Embase) through July/09/2021 and identified 33 studies reporting associations with prenatal organochlorine exposure, 21 studies reporting associations with prenatal PFAS, and five studies reporting associations with prenatal PBDEs. We conducted a qualitative review. Additionally, we performed random-effects meta-analyses of POP exposures, with data estimates from at least three prospective studies, and BMI-z. Prenatal DDE and HCB levels were associated with higher BMI z-score in childhood (beta: 0.12, 95% CI: 0.03, 0.21; I2 : 28.1% per study-specific log increase of DDE and beta: 0.31, 95% CI: 0.09, 0.53; I2 : 31.9% per study-specific log increase of HCB). No significant associations between PCB-153, PFOA, PFOS, or pentaPBDEs with childhood BMI were found in meta-analyses. In individual studies, there was inconclusive evidence that POP levels were positively associated with other obesity indicators (e.g., waist circumference).


Upper and lower rib cage deformation during breathing in quadriplegics.
In quadriplegia, the anteroposterior diameter of the rib cage (RC) decreases during inspiration. Our aim was to characterize this paradoxical motion further and to determine to what extent it was due to a diminished inspiratory effect of the diaphragm on the RC because of abdominal muscle paralysis. In six quadriplegics, upper and lower RC (URC, LRC) displacements were studied in various postures with and without extrinsic abdominal support. LRC was in its relaxed configuration at all lung volumes studied in three subjects before and four subjects during abdominal compression. URC distorted most and, despite improvement during abdominal compression, was never in the relaxed configuration during tidal inspiration. Thoracic distortability was further studied by noting the effect on URC and LRC of a cephalad force applied to the RC at the costal margin. This produced distortion similar to that observed during spontaneous inspiration. It was concluded that during spontaneous inspiration in quadriplegics lack of abdominal support does not entirely explain the RC distortion which occurs, and high abdominal pressure results in diminished RC distortion, an effect which we attribute to an increase in the area of diaphragmatic apposition to the RC.


Decoding "us" and "them": Neural representations of generalized group concepts.
Humans form social coalitions in every society on earth, yet we know very little about how the general concepts us and them are represented in the brain. Evolutionary psychologists have argued that the human capacity for group affiliation is a byproduct of adaptations that evolved for tracking coalitions in general. These theories suggest that humans possess a common neural code for the concepts in-group and out-group, regardless of the category by which group boundaries are instantiated. The authors used multivoxel pattern analysis to identify the neural substrates of generalized group concept representations. They trained a classifier to encode how people represented the most basic instantiation of a specific social group (i.e., arbitrary teams created in the lab with no history of interaction or associated stereotypes) and tested how well the neural data decoded membership along an objectively orthogonal, real-world category (i.e., political parties). The dorsal anterior cingulate cortex/middle cingulate cortex and anterior insula were associated with representing groups across multiple social categories. Restricting the analyses to these regions in a separate sample of participants performing an explicit categorization task, the authors replicated cross-categorization classification in anterior insula. Classification accuracy across categories was driven predominantly by the correct categorization of in-group targets, consistent with theories indicating in-group preference is more central than out-group derogation to group perception and cognition. These findings highlight the extent to which social group concepts rely on domain-general circuitry associated with encoding stimuli's functional significance. (PsycINFO Database Record


Lesioning of the hypothalamic paraventricular nucleus inhibits ether-induced ACTH but not prolactin release.
Stress mediators, CRF-41 and vasopressin known to be synthesized in, and released from the parvocellular neurosecretory neurons of the hypothalamic paraventricular nucleus (PVN) are essential to release adrenocorticotropin (ACTH) in response to stress. In addition, suckling-induced prolactin (PRL) release also depends on the integrity of the PVN. In the present study, ether stress-induced adrenocorticotrop hormone (ACTH) and prolactin (PRL) release was studied 2, 5 and 42 days after placing lesions in the hypothalamic paraventricular nucleus (PVN) of male rats. Ether-induced ACTH secretion was strongly inhibited 2 and 5 days after lesions whereas 6 weeks later the lesion induced inhibition was fading. In contrast, PVN lesion failed to inhibit ether-induced PRL release at any time studied. The results suggest that contrary to previous suggestions the peptidergic neurons essential for stress-induced PRL release are outside the PVN.


A methodological approach to non-invasive assessments of vascular function and morphology.
The endothelium is the innermost lining of the vasculature and is involved in the maintenance of vascular homeostasis. Damage to the endothelium may predispose the vessel to atherosclerosis and increase the risk for cardiovascular disease. Assessments of peripheral endothelial function are good indicators of early abnormalities in the vascular wall and correlate well with assessments of coronary endothelial function. The present manuscript details the important methodological steps necessary for the assessment of microvascular endothelial function using laser Doppler imaging with iontophoresis, large vessel endothelial function using flow-mediated dilatation, and carotid atherosclerosis using carotid artery ultrasound. A discussion on the methodological considerations for each of the techniques is also presented, and recommendations are made for future research.


An iron-regulated ferric reductase associated with the absorption of dietary iron.
The ability of intestinal mucosa to absorb dietary ferric iron is attributed to the presence of a brush-border membrane reductase activity that displays adaptive responses to iron status. We have isolated a complementary DNA, Dcytb (for duodenal cytochrome b), which encoded a putative plasma membrane di-heme protein in mouse duodenal mucosa. Dcytb shared between 45 and 50% similarity to the cytochrome b561 family of plasma membrane reductases, was highly expressed in the brush-border membrane of duodenal enterocytes, and induced ferric reductase activity when expressed in Xenopus oocytes and cultured cells. Duodenal expression levels of Dcytb messenger RNA and protein were regulated by changes in physiological modulators of iron absorption. Thus, Dcytb provides an important element in the iron absorption pathway.


Consideration of cytokines as therapeutics agents or targets.
There has been an explosion of our knowledge in cytokine biology in the last decade. Such knowledge is being quickly translated into the identification of etiologies and improved prophylaxis and therapy of disease. While cytokines have the potential to be used as therapeutics or immune adjuvants for certain diseases, they may also be culprits as therapeutic targets in other diseases. This review article serves, as an introduction to the other five articles in this thematic issue each of which has a specific focus on the frontier of cytokine therapeutic biology. This review contains sections dealing with general cytokine properties, cytokine classifications, human conditions caused by cytokine under-expression and over-expression, Th1 and Th2 paradigm, cytokine therapy for acute/chronic inflammatory conditions, cytokine therapy for infectious diseases, and cytokine therapy for cancer.


Nongastric marginal-zone B-cell MALT lymphoma: prognostic value of disease dissemination.
The aim of this study is to describe the clinical features and define the prognostic significance of disease dissemination in a large series of nongastric marginal-zone B-cell mucosa-associated lymphoid tissue (MALT) lymphomas. We studied 208 patients with nongastric marginal-zone B-cell MALT lymphoma diagnosed and treated from 1991 to 2004. Ninety percent of the patients had a single site of MALT involvement--skin (26%), salivary glands (18%), orbit (14%), Waldeyer's ring (13%)--and 39% and 28% had nodal involvement and bone marrow involvement, respectively. After a median follow-up of 2.7 years, the median event-free survival (EFS) time was 2.4 years, and the median overall survival (OS) time was not reached. On univariate analysis, the features significantly associated with longer EFS and OS times were the following: single MALT site involvement (OS), localized disease (EFS and OS), no nodal disease (EFS and OS), skin and orbit lymphoma (OS), and stage IV disease without bone marrow involvement (OS). On multivariate analysis, both bone marrow and nodal involvement were associated with shorter OS. This study describes the clinical features and the natural history of nongastric marginal-zone lymphomas and highlights that the dissemination to lymph nodes and bone marrow is associated with a poorer outcome.


Emergency department chief complaint and diagnosis data to detect influenza-like illness with an electronic medical record.
The purpose of syndromic surveillance is early detection of a disease outbreak. Such systems rely on the earliest data, usually chief complaint. The growing use of electronic medical records (EMR) raises the possibility that other data, such as emergency department (ED) diagnosis, may provide more specific information without significant delay, and might be more effective in detecting outbreaks if mechanisms are in place to monitor and report these data. The purpose of this study is to characterize the added value of the primary ICD-9 diagnosis assigned at the time of ED disposition compared to the chief complaint for patients with influenza-like illness (ILI). The study was a retrospective analysis of the EMR of a single urban, academic ED with an annual census of over 60, 000 patients per year from June 2005 through May 2006. We evaluate the objective in two ways. First, we characterize the proportion of patients whose ED diagnosis is inconsistent with their chief complaint and the variation by complaint. Second, by comparing time series and applying syndromic detection algorithms, we determine which complaints and diagnoses are the best indicators for the start of the influenza season when compared to the Centers for Disease Control regional data for Influenza-Like Illness for the 2005 to 2006 influenza season using three syndromic surveillance algorithms: univariate cumulative sum (CUSUM), exponentially weighted CUSUM, and multivariate CUSUM. In the first analysis, 29% of patients had a different diagnosis at the time of disposition than suggested by their chief complaint. In the second analysis, complaints and diagnoses consistent with pneumonia, viral illness and upper respiratory infection were together found to be good indicators of the start of the influenza season based on temporal comparison with regional data. In all examples, the diagnosis data outperformed the chief-complaint data. Both analyses suggest the ED diagnosis contains useful information for detection of ILI. Where an EMR is available, the short time lag between complaint and diagnosis may be a price worth paying for additional information despite the brief potential delay in detection, especially considering that detection usually occurs over days rather than hours.


[DNA content and synthesis in the nuclei of rat cerebellar cells in organotypic cultures].
Using cytophotometric and autoradiographic methods, it has been shown for the first time that in condition of an organotypic culture the replicative synthesis of DNA is induced in the Purkinje neurons of the cerebellum of newborn rats completing their terminal differentiation. This synthesis is accompanied by polyploidization of the initially diploid population of these cells (4c, much more rarely 8c, and a single 16c cell appear) rather than by cell division. In constant, the granular cells mostly retain their diploid state and only a few of them synthesize DNA to H2c values. The glial cells divide actively. Hence, evidence is presented that neurons, at least those of cerebellum, retain their potential of replicative synthesis of DNA in the organotypic culture. The important point is that DNA synthesis in their nuclei proceeds simultaneously with processes of differentiation.


Arthroscopic treatment of ankle impingement syndrome.
Arthroscopic treatment of ankle impingement syndrome (AIS) is a minimally invasive surgical procedure used to address symptoms caused by impingement in the ankle joint. This syndrome occurs when there is abnormal contact between certain bones or soft tissues in the ankle, leading to pain, swelling, or limited range of motion. Traditionally, open surgery was the standard approach for treating AIS. However, with advancements in technology and surgical techniques, arthroscopic treatment has become a preferred method for many patients and surgeons. With improved visualization and precise treatment of the arthroscopy, patients can experience reduced pain and improved functionality, allowing them to return to their daily activities sooner. In this paper, we reviewed the application and clinical efficacy the of arthroscopic approach for treating AIS, hoping to provide a reference for its future promotion.


Relationship between the calcium-mobilizing action of inositol 1,4,5-trisphosphate in permeable AR4-2J cells and the estimated levels of inositol 1,4,5-trisphosphate in intact AR4-2J cells.
Various experimental strategies were employed in an effort to explain the previously reported [Horstman, Takemura & Putney (1988) J. Biol. Chem. 263, 15297-15303] paradoxically high levels of inositol 1,4,5-trisphosphate [(1,4,5)IP3] in resting and substance-P-stimulated AR4-2J cells. The concentration-effect curves for substance-P-induced [3H](1,4,5)IP3 formation in [3H]inositol-labelled cells and substance-P-induced increase in intracellular [Ca2+] were essentially superimposable, suggesting that formation of (1,4,5)IP3 is limiting for cellular Ca2+ mobilization. In electrically permeabilized AR4-2J cells, (1,4,5)IP3 and other inositol polyphosphates stimulated Ca2+ release with potencies similar to those reported for other cell types, including the parent pancreatic acinar cell. Compartmentalization of basal (1,4,5)IP3 was suggested by the fact that this material was stable in the presence of antimycin A, although this toxin completely blocked agonist stimulation of phospholipase C. However, subcellular fractionation as well as permeabilization of the cells with Staphylococcus aureus alpha-toxin failed to provide evidence for binding or sequestration of [3H](1,4,5)IP3 in AR4-2J cells. The density of (1,4,5)IP3 receptors in AR4-2J cells was not sufficiently large to impose non-linearity in the relationship between (1,4,5)IP3 concentration and (1,4,5)IP3-induced Ca2+ release. Thus the apparent high concentrations of (1,4,5)IP3 in resting and stimulated AR4-2J cells are not indicative of atypically low sensitivity or high concentration of (1,4,5)IP3 receptors, nor is there evidence for compartmentalization of (1,4,5)IP3 outside of the cytoplasm in these cells. It is possible that soluble factors in the cytoplasm of AR4-2J cells regulate the free concentration of (1,4,5)IP3 or the sensitivity of receptors to (1,4,5)IP3.


Small intestinal permeability as an indicator of jejunal mucosal recovery in patients with celiac sprue on a gluten-free diet.
Lactulose/mannitol excretion ratios were measured in 13 patients with celiac disease at diagnosis and after 5-8 months on a gluten-free diet. Jejunal biopsies were assessed histologically at diagnosis and during treatment. The excretion ratios in untreated patients were significantly higher than in 25 normal controls (P less than 0.01). On the diet, the excretion ratios fell in every patient, but in only eight did the ratio return to normal. There was a good correlation between the ratio and jejunal histological grading. During treatment, the ratios significantly inversely correlated with jejunal villous height/mucosal thickness ratios (P less than 0.001). Therefore, excretion ratios provide a well-tolerated noninvasive means of assessing the jejunal mucosa in patients with celiac disease on a gluten-free diet.


A Simple Method for Quantifying Larval Locomotion in Drosophila melanogaster.
The fruit fly Drosophila melanogaster is a powerful genetic model that has been used for many decades to study nervous system function, development, and behavior. There are a large number of developmental and behavioral traits that can be measured to provide a broad readout of neurological function. These include patterned motor behaviors, such as larval locomotion, which can be used to assess whether genetic or environmental factors affect nervous system function to provide an entry point for deeper mechanistic studies. Here, we describe a protocol for quantifying larval locomotion using a simple camera setup and a freely available image analysis software. This protocol can be readily applied to human disease models or in toxicology studies, for example, to broadly assess the impact of treatments on neurological function.


Positive end-expiratory pressure and renal function influence B-type natriuretic peptide in patients with severe sepsis and septic shock.
Myocardial dysfunction is a complication associated with a poor prognosis in septic patients. A biomarker of cardiac function providing prognostic information is of paramount interest. We sought to determine the value of B-type natriuretic peptide in patients with severe sepsis/septic shock. We performed a prospective study in patients with severe sepsis/septic shock in a medical intensive care unit. B-type natriuretic peptide level was determined within 24 hours after the diagnosis of severe sepsis/septic shock. We also analyzed mortality, and presence of association between B-type natriuretic peptide and clinical, hemodynamic and respiratory variables. 23 (9 women; 14 men) patients with ages ranging from 20-79 (mean 51.3+/-18.6) years old and APACHE score of 22.6+/-11.8 were included; 15 (65.2%) patients received pulmonary artery catheters, and 20 (87%) were mechanically ventilated. Multivariate analysis disclosed inverse association between B-type natriuretic peptide values with positive end-expiratory pressure values, and direct association with creatinine (beta 0.548 and 0.377, p 0.02 and 0.002, respectively), but not with mortality, clinical and hemodynamic parameters. This is the first report on an inverse association between positive end-expiratory pressure and BNP levels in patients with severe sepsis and septic shock. BNP and creatinine levels should be taken into consideration when analyzing B-type natriuretic peptide levels in this setting.


Microvasculature in murine tracheal allografts after combined therapy with clopidogrel and everolimus.
Survival after lung transplantation is mainly limited by the development of chronic lung allograft dysfunction. Previous studies have suggested T-cell mediated proliferation and microvascular changes in experimental small airways models as potential therapeutic targets. The aim of this study was to assess microvascular changes in murine orthotopic tracheal allografts after treatment with everolimus alone or in combination with clopidogrel. C57Bl/6 (H-2b) donor tracheas were orthotopically transplanted into CBA (H-2k) recipients. Mice received daily injections of everolimus (0.05 mg/kg) alone or combined with clopidogrel (1 mg/kg). Twenty-eight days after transplantation, ratio of the thickness of tracheal epithelium and lamina propria was measured as an indicator for chronic rejection. Additionally, graft oxygenation and graft perfusion were detected on postoperative days 4, 10 and 28. Quantitative reverse transcription polymerase chain reaction analysis was used for gene expression analysis. While syngeneic grafts showed a stable tissue pO2 and undisturbed microvascular perfusion, rejecting allografts had a drastic decline in both parameters as well as a flattened epithelium and an increased thickness of the lamina propria. Treatment with everolimus reduced allogeneic fibroproliferation, but had no protective effects on the microvasculature; polymerase chain reaction analysis indicated hypoxic stress and inflammation. Combining everolimus with clopidogrel improved microvascular integrity in the tracheal grafts, but had no synergistic effect in preventing obliterative bronchiolitis development. These data demonstrate that the ability of everolimus to reduce the development of post-transplant obliterative bronchiolitis is not caused by microvascular protection and has no synergistic effects with clopidogrel in acute airway rejection.


Differentiation of BCMA-specific induced pluripotent stem cells into CD8αβ+ T cells targeting multiple myeloma.
A major hurdle in adoptive T cell therapy is cell exhaustion and failure to maintain anti-tumor responses. Here, we introduce an induced pluripotent stem cell (iPSC) strategy for reprogramming and revitalization of precursor exhausted BCMA-specific T cells to effectively target multiple myeloma (MM). Heteroclitic BCMA72-80 (YLMFLLRKI)-specific CD8+ memory cytotoxic T lymphocytes (CTL) were epigenetically reprogrammed to a pluripotent state, developed into hematopoietic progenitor cells (HPC: CD34+ CD43+/CD14- CD235a-), differentiated into the T cell lineage and evaluated for their poly-functional activities against MM. The final T cell products demonstrated; 1) mature CD8αβ+ memory phenotype, 2) high expression of activation/costimulatory molecules (CD38, CD28, 41BB), 3) no expression of immune checkpoint and senescence markers (CTLA4, PD1, LAG3, TIM3; CD57), and 4) robust proliferation and poly-functional immune responses to MM. The BCMC-specific iPSC-T cells possessed a single T cell receptor clonotype with cognate BCMA peptide recognition and specificity for targeting MM. RNAseq analyses revealed distinct genome-wide shifts and a distinctive transcriptional profile in selected iPSC clones, which can develop CD8αβ+ memory T cells. This includes a repertoire of gene regulators promoting T cell lineage-development, memory CTL activation, and immune response regulation [LCK, IL7R; 4-1BB, TRAIL, GZMB, FOXF1, ITGA1]. This study highlights the potential application of iPSC technology to an adaptive T cell therapy protocol and identifies specific transcriptional patterns that could serve as a biomarker for selection of suitable iPSC clones for successful development of antigen-specific CD8αβ+ memory T cells to improve MM patient outcome.


Scale-Up Technologies for the Manufacture of Adherent Cells.
Great importance is being given to the impact our food supply chain and consumers' food habits are having on the environment, human health, and animal welfare. One of the latest developments aiming at positively changing the food ecosystem is represented by cultured meat. This form of cellular agriculture has the objective to generate slaughter-free meat products starting from the cultivation of few cells harvested from the animal tissue of interest. As a consequence, a large number of cells has to be generated at a reasonable cost. Just to give an idea of the scale, there were billions of cells just in a bite of the first cultured-meat burger. Thus, one of the major challenges faced by the scientists involved in this new ambitious and fascinating field, is how to efficiently scale-up cell manufacture. Considering the great potential presented by cultured meat, audiences from different backgrounds are very interested in this topic and eager to be informed of the challenges and possible solutions in this area. In light of this, we will provide an overview of the main existing bioprocessing technologies used to scale-up adherent cells at a small and large scale. Thus, giving a brief technical description of these bioprocesses, with the main associated advantages and disadvantages. Moreover, we will introduce an alternative solution we believe has the potential to revolutionize the way adherent cells are grown, helping cultured meat become a reality.


[Cannabis in oncology - much ado about nothing?]
The medical use of Cannabis has gained popularity in Europe and Northern America in recent years. Cannabinoids are available as finished pharmaceuticals, flowers and extracts. This article focuses on supportive medicine for oncological patients. Possible indications are pain, chemotherapy-induced nausea and vomiting, loss of appetite and altered taste perception. Despite the enormous cannabis hype in medicine, the evidence for its use in oncology patients is insufficient. However, palliative patients with refractory symptoms could be candidates for a therapeutic trial. The key parameter for choosing a cannabis medicinal product is the THC/CBD ratio. Oral forms of administration are particularly suitable for cannabis-naive and older patients. Mental and cardiovascular side effects should not be underestimated.


Changing Trends in Surgical Management of Prostate Cancer: The End of Overtreatment?
The use of prostate-specific antigen (PSA) for screening or early detection of prostate cancer (PCa) results in significant stage migration toward more favorable stages and a proven decrease in PCa mortality but is accompanied by substantial rates of overdiagnosis and overtreatment. Acknowledgement of these downsides and endeavors to avoid them have led to substantial changes in treatment patterns. Many centers have reported dramatic changes, with increases in active surveillance (AS) of early cancers and local treatment of advanced disease. To estimate the impact of this development on our radical prostatectomy (RP) series, we analyzed changes in cancer and patient selection over the past 15 yr. Despite a trend toward decreased utilization of RP in Germany, the annual caseload at our institution increased due to regionalization, from 382 RPs in 2000 to 2145 in 2011, and has been stable for the past 3 yr (2106 RPs in 2014). The rate of RPs performed in patients with low-risk PCa, AS candidates, or men with a pure Gleason 6 pattern in the RP specimen dropped from 60%, 38.2%, and 56.2%, respectively, in 2004 to 27%, 14.7%, and 10%, respectively, in 2011-2013. Patients undergoing RP with solely Gleason 6 cancer were younger on average (aged 61 yr) than patients in higher risk groups (aged 65 yr). The rate of histologically insignificant PCa was low, ranging from 1% to 8.8% depending on the definition used. Patient selection is the other important tool used to avoid overtreatment. Long-term other-cause mortality (OCM) should be low in adequately selected RP candidates, and after a minimum follow-up of 15 yr, overall OCM was 14.8%. The OCM rate was 10.2% in men aged <65 yr and 24.3% in men aged ≥65 yr. The current analysis documents a clear shift in utilization of RP toward significant PCa in men with long life expectancy. Based on patient and cancer selection as described, the long-standing discussion of overtreatment with RP might become invalid. Discussion of possible overtreatment has led to dramatic changes in indication for radical prostatectomy (RP). We analyzed a large European patient cohort and found that RP is rarely done in early cancers but is used more for aggressive tumors. Those who underwent RP had long life expectancy and benefit from surgery. With this change in application, overtreatment with RP is unlikely.


Continuity of Care and Self-Management among Patients with Stroke: A Cross-Sectional Study.
(1) Background: Stroke is an important topic in the healthcare industry. The objective of the present study was to investigate patients' sociodemographic characteristics, health status, continuity of care, self-management, and other predictors that affect their self-management. (2) Methods: This cross-sectional correlational study was carried out from March to September 2020, and included a total of 150 patients aged 20 and above who were diagnosed within the past 6 months. The research participants were selected from the Division of Neurology, Department of Internal Medicine/Department of Surgery, at a medical center in Central Taiwan. (3) Results: The mean self-management score of patients with stroke was 110.50 points (30-150 points). As shown in the stepwise regression analysis, the overall regression model explained approximately 44.5% of the variance in self-management. Educational level (10.8%), frequency of exercise per week (2.1%), time that patients were affected by stroke (2.4%), and continuity of care (29.2%) were the main predictors affecting the self-management of stroke patients. (4) Conclusions: To improve stroke patients' self-management, medical teams should provide appropriate continuity of care to those with lower educational levels, those without exercise habits, and those who experienced a stroke within the past six months.


EGF and HB-EGF modulate inward potassium current in human bladder urothelial cells from normal and interstitial cystitis patients.
Interstitial cystitis (IC) is an idiopathic condition characterized by bladder hyperalgesia. Studies have shown cytokine and purinergic signaling abnormalities in cultured bladder urothelial cells (BUC) from IC patients. We performed single-cell electrophysiological studies in both normal and IC BUC. A strongly inward rectifying potassium current with conductance of the Kir2.1 channel was identified in normal BUC. This current was significantly reduced in IC BUC. Kir2.1 protein and mRNA were detected in both IC and normal BUC. Epidermal growth factor (EGF) caused a dose-dependent decrease in the inward potassium current in normal BUC. EGF is secreted in higher amounts by IC BUC and is known to decrease Kir2.1 conductance by phosphorylation of Kir2.1. Genistein, a nonspecific phosphorylation inhibitor, increased the inward potassium current in IC BUC and blocked the effect of EGF on normal BUC. Treatment of IC BUC with heparin-binding epidermal growth factor-like growth factor (HB-EGF), previously shown to be secreted in lower amounts by IC BUC, significantly increased inward potassium current. These data show that the inward potassium current in BUC can be modulated by EGF and HB-EGF. Changes in BUC membrane potassium conductance caused by altered levels of EGF and HB-EGF may therefore play a role in the pathophysiology of IC.


Nano-structure of the laminin γ-1 short arm reveals an extended and curved multidomain assembly.
Laminins are multidomain glycoproteins that play important roles in development and maintenance of the extracellular matrix via their numerous interactions with other proteins. Several receptors for the laminin short arms revealed their importance in network formation and intercellular signaling. However, both the detailed structure of the laminin γ-1 short arm and its organization within the complexes is poorly understood due to the complexity of the molecule and the lack of a high-resolution structure. The presented data provide the first subatomic resolution structure for the laminin γ-1 short arm in solution. This was achieved using an integrated approach that combined a number of complementary biophysical techniques such as small angle X-ray scattering (SAXS), analytical ultracentrifugation, dynamic light scattering and electron microscopy. As a result of this study, we have obtained a significantly improved model for the laminin γ-1 short arm that represents a major step forward in molecular understanding of laminin-mediated complex formations.


Government supervision on quality of smoking-cessation counselling in midwifery practices: a qualitative exploration.
The Dutch Healthcare Inspectorate supervises care providers in order to improve quality of care. Recently the inspectorate assessed and promoted the use of a guideline on smoking-cessation counselling in midwifery practices. The supervision programme consisted of an announcement of the enforcement deadline for the guideline and site visits. The purpose of our qualitative study was to identify factors related to guideline adherence after the supervision programme, and investigate whether the programme had helped improve adherence. We conducted semi-structured interviews with inspected and non-inspected midwives. Additionally, we studied documents and observed the inspection process. The sampled midwives all work in primary care midwifery practices providing care to pregnant smokers. The questions included the current provision of smoking-cessation counselling, support to the midwife in counselling, recent changes in provision of counselling, reasons for recent changes, knowledge about the supervision programme, and experiences with supervision by the inspectorate. Our results show that guideline adherence depends on several factors. Awareness and familiarity with the guideline are important, as is outcome expectancy. Additionally, motivation, guideline factors and environment factors were mentioned. Besides these previously documented factors, we found that professional collaboration also determined guideline adherence. Increased collaboration in counselling is associated with greater adherence to the guideline, such as provision of counselling and taking required training. The supervision programme helped improve stop-smoking counselling, by making midwives aware of the counselling and giving them an extrinsic motivation to provide counselling. Motivation and environmental aspects were the most important factors related to guideline adherence, and professional environment was added as significant factor. The improved guideline adherence is partly attributable to the supervision programme.


Conjugated linoleic acid content of milk and cheese from cows fed extruded oilseeds.
Extruded oilseeds were fed to 24 dairy cows to study the influence on the conjugated linoleic acid content of milk and cheese. Cows were fed one of three diets that contained forage and grain in a ratio of 47:53. A control diet containing 13.5% soybean meal was compared with diets containing 12% full fat extruded soybeans or 12% full fat extruded cottonseed. The control, extruded soybean, and extruded cottonseed diets contained 2.73, 4.89, and 4.56% fatty acids, respectively. Measurements were made during the last 5 wk of the 8-wk experiment. The DM intakes and 3.5% fat-corrected milk yields were higher for cows fed the extruded soybean and extruded cottonseed diets than for cows fed the control diet. A tendency for lower fat and protein contents in the milk of cows fed the extruded soybean and extruded cottonseed diets was detected. Most of the C18 fatty acids were increased in the milk and cheese when extruded soybeans and cottonseeds were fed. The conjugated linoleic acid content in milk and cheese increased a mean of 109% when full fat extruded soybeans were fed and increased 77% when cottonseeds were fed compared with the conjugated linoleic acid content when the control diet was fed. Processing the milk into cheese did not alter the conjugated linoleic acid content. The conjugated linoleic acid content of milk and cheese can be increased by the inclusion of full fat extruded soybeans and full fat extruded cottonseeds in the diets of dairy cows.


Ability of iron to promote surfactant peroxide decomposition and oxidize alpha-tocopherol.
Peroxides are an important factor in oxidative reactions in foods because their decomposition can result in formation of highly reactive free radicals. Emulsifiers such as the Brijs, Tweens, and lecithin were found to contain 4-35 micromol of peroxides/g of surfactant. Peroxide concentrations in Tween 20 micelles increased in the presence of low iron concentrations but decreased when iron concentrations were high, suggesting that iron was capable of promoting both peroxide formation and decomposition. Oxidation of alpha-tocopherol was observed in micelles high in peroxides (Tween 20) but not in micelles where peroxide concentrations were low (Brij). Transition metals accelerated the oxidation of alpha-tocopherol in Tween 20 micelles, whereas EDTA stabilized alpha-tocopherol in the presence of added Fe(2+). These results suggest that surfactant peroxides could decrease the oxidative stability of food emulsions by acting as a source of free radicals, especially in the presence of transition metals.


Comparative effectiveness of digital versus film-screen mammography in community practice in the United States: a cohort study.
Few studies have examined the comparative effectiveness of digital versus film-screen mammography in U.S. community practice. To determine whether the interpretive performance of digital and film-screen mammography differs. Prospective cohort study. Mammography facilities in the Breast Cancer Surveillance Consortium. 329,261 women aged 40 to 79 years underwent 869 286 mammograms (231 034 digital; 638 252 film-screen). Invasive cancer or ductal carcinoma in situ diagnosed within 12 months of a digital or film-screen examination and calculation of mammography sensitivity, specificity, cancer detection rates, and tumor outcomes. Overall, cancer detection rates and tumor characteristics were similar for digital and film-screen mammography, but the sensitivity and specificity of each modality varied by age, tumor characteristics, breast density, and menopausal status. Compared with film-screen mammography, the sensitivity of digital mammography was significantly higher for women aged 60 to 69 years (89.9% vs. 83.0%; P = 0.014) and those with estrogen receptor-negative cancer (78.5% vs. 65.8%; P = 0.016); borderline significantly higher for women aged 40 to 49 years (82.4% vs. 75.6%; P = 0.071), those with extremely dense breasts (83.6% vs. 68.1%; P = 0.051), and pre- or perimenopausal women (87.1% vs. 81.7%; P = 0.057); and borderline significantly lower for women aged 50 to 59 years (80.5% vs. 85.1%; P = 0.097). The specificity of digital and film-screen mammography was similar by decade of age, except for women aged 40 to 49 years (88.0% vs. 89.7%; P < 0.001). Statistical power for subgroup analyses was limited. Overall, cancer detection with digital or film-screen mammography is similar in U.S. women aged 50 to 79 years undergoing screening mammography. Women aged 40 to 49 years are more likely to have extremely dense breasts and estrogen receptor-negative tumors; if they are offered mammography screening, they may choose to undergo digital mammography to optimize cancer detection. National Cancer Institute.


Polywater: an attempt at synthesis in a gas discharge.
An attempt to produce polywater in a corona discharge in moist air was unsuccessful. However, the major product produced, nitric acid, has a midrange infrared spectrum which is strikingly similar to that reported for polywater. The Raman spectrum offers a better means of distinguishing between nitric acid and polywater than the infrared spectrum does.


Genotyping and haplotyping of CYP2C19 functional alleles on thin-film biosensor chips.
Numerous functional polymorphisms in the CYP2C19 gene have been identified; some alleles (e.g. CYP2C19*2 and CYP2C19*3) are associated with poor metabolism of CYP2C19 substrate drugs. Studies have found that the proportion of poor metabolizers, explained by CYP2C19*2 and CYP2C19*3, varies from less than 50% to more than 90% of poor metabolizers. Therefore, phenotype-genotype correlation studies should cover more than CYP2C19*2 and CYP2C19*3. A broader coverage, however, requires an easy-to-use and high-throughput genotyping platform. This broader coverage should also include the recently identified functional allele, CYP2C19*10, which involves a nucleotide change adjacent to the altered nucleotide change in CYP2C19*2. The currently used restriction fragment length polymorphism-based method for genotyping CYP2C19*2 cannot distinguish between CYP2C19*2 and CYP2C19*10. We aim to develop a simple platform that can genotype all CYP2C19 functional alleles. We have developed a thin-film biosensor chip platform to genotype 16 exonic CYP2C19 variants, including two sets of two adjacent single nucleotide polymorphisms and 12 single single nucleotide polymorphisms, using a ligation strategy. We demonstrate that this is a rapid, accurate, and inexpensive method for genotyping CYP2C19 variants using individual's genomic DNA samples. We further demonstrate that this genotyping platform can be used to construct a haplotype structure of the CYP2C19 variants in a population, and to assign a haplotype combination to each individual on the basis of his/her genotype results. This assay can be applied in pharmacogenomic studies in both basic research and clinical laboratories. It is also an ideal technology for pharmacogenomic tests in both developed and developing countries.


Strategies to discover the structural components of cyst and oocyst walls.
Cysts of Giardia lamblia and Entamoeba histolytica and oocysts of Toxoplasma gondii and Cryptosporidium parvum are the infectious and sometimes diagnostic forms of these parasites. To discover the structural components of cyst and oocyst walls, we have developed strategies based upon a few simple assumptions. Briefly, the most abundant wall proteins are identified by monoclonal antibodies or mass spectrometry. Structural components include a sugar polysaccharide (chitin for Entamoeba, β-1,3-linked glucose for Toxoplasma, and β-1,3-linked GalNAc for Giardia) and/or acid-fast lipids (Toxoplasma and Cryptosporidium). Because Entamoeba cysts and Toxoplasma oocysts are difficult to obtain, studies of walls of nonhuman pathogens (E. invadens and Eimeria, respectively) accelerate discovery. Biochemical methods to dissect fungal walls work well for cyst and oocyst walls, although the results are often unexpected. For example, echinocandins, which inhibit glucan synthases and kill fungi, arrest the development of oocyst walls and block their release into the intestinal lumen. Candida walls are coated with mannans, while Entamoeba cysts are coated in a dextran-like glucose polymer. Models for cyst and oocyst walls derive from their structural components and organization within the wall. Cyst walls are composed of chitin fibrils and lectins that bind chitin (Entamoeba) or fibrils of the β-1,3-GalNAc polymer and lectins that bind the polymer (Giardia). Oocyst walls of Toxoplasma have two distinct layers that resemble those of fungi (β-1,3-glucan in the inner layer) or mycobacteria (acid-fast lipids in the outer layer). Oocyst walls of Cryptosporidium have a rigid bilayer of acid-fast lipids and inner layer of oocyst wall proteins.


Nematicidal Activity of the Volatilome of Eruca sativa on Meloidogyne incognita.
Research on new pesticides based on plant extracts, aimed at the development of nontoxic formulates, has recently gained increased interest. This study investigated the use of the volatilome of rucola (Eruca sativa) as a powerful natural nematicidal agent against the root-knot nematode, Meloidogyne incognita. Analysis of the composition of the volatilome, using GC-MS-SPME, showed that the compound (Z)-3-hexenyl acetate was the most abundant, followed by (Z)-3-hexen-1-ol and erucin, with relative percentages of 22.7 ± 1.6, 15.9 ± 2.3, and 8.6 ± 1.3, respectively. Testing of the nematicidal activity of rucola volatile compounds revealed that erucin, pentyl isothiocyanate, hexyl isothiocyanate, (E)-2-hexenal, 2-ethylfuran, and methyl thiocyanate were the most active with EC50 values of 3.2 ± 1.7, 11.1 ± 5.0, 11.3 ± 2.6, 15.0 ± 3.3, 16.0 ± 5.0, and 18.1 ± 0.6 mg/L, respectively, after 24 h of incubation. Moreover, the nematicidal activity of fresh rucola used as soil amendant in a containerized culture of tomato decreased the nematode infection in a dose-response manner (EC50 = 20.03 mg/g) and plant growth was improved. On the basis of these results, E. sativa can be considered as a promising companion plant in intercropping strategies for tomato growers to control root-knot nematodes.


Eruptive malpositioning of the mandibular permanent lateral incisors: three case reports.
Ectopic eruption of the mandibular permanent lateral incisors involving the loss of the adjacent primary canines is not uncommon, yet the extreme distalization and transposition of such teeth (partial or true) has been reported infrequently. Three cases that illustrate various anomalous eruption-exfoliation patterns are presented. Theories that attempt to explain the etiology of anomalous eruption positions are discussed. However, the multifactorial process of growth and development makes it difficult to identify specific primary etiological factors. Treatment options also are discussed and illustrated, with emphasis on early orthodontic intervention for optimal results.


Revisited Surgical Anatomy of the Left Colonic Angle for Tailored Carcinologic Colectomy: A Review.
Although several types of surgical procedure have been advocated to date, the optimal resection of the left colonic angle in cancer treatment remains controversial. Located at the border of the transverse and descending colons, the anatomy of the left colonic angle is complex and characterized by numerous anatomic variations. Recent advances in preoperative (three-dimensional CT angiography with colonography) and/or intraoperative (indocyanine green staining) imaging have allowed for a better identification of these variations. We performed a methodological review of studies assessing the anatomical variations of the left colic artery. While the left colonic angle is classically vascularized by branches of the superior and inferior mesenteric arteries, an accessory middle colonic artery has been identified from 6 % to 36% of cases, respectively, leading to their classification of five types. In the absence of a left colic artery, this artery becomes predominant. In parallel to the variations in the venous drainage of the left colonic angle, which has been classified into four types, new lymphatic drainage routes have also been identified via this accessory artery and the inferior mesenteric vein. Collectively, these newly obtained findings plead for preoperative identification in cases of cancer of the left colonic angle and a surgical strategy adapted to these anatomical variations.


Prevalence of musculoskeletal pain in two primary care clinics in a mid-sized town's urban population in Malaysia.
This study assesses the prevalence of musculoskeletal (MSK) pain in patients attending primary care clinics in a medium-sized town in Malaysia and examines the interventions given for the symptoms and the level of the associated disabilities. This investigation comprises a cross-sectional descriptive study of all patients visiting two primary care clinics aged 18 years and above. Patients presenting with joint pain answered a questionnaire assessing demographic data, disabilities (measured by the Stanford HAQ-DI), and treatment options. Of 1,074 patients surveyed, 202 (18.8%) had MSK complaints. The mean age of those with MSK pain was 56.1 years. Incidence increased with age, reaching 78.8% of those over 48 years of age. The knee was the most common site of MSK pain (52.2%), with 20.3% requiring referral for specialist assessment. The median HAQ score was 0.375 and 89.6% of those surveyed had mild disability. MSK pain is a common problem among patients visiting primary care clinics. The most common site of MSK pain was the knee. On formal assessment, the majority of these patients exhibited mild disability. A significant proportion of patients still required specialist referral. This finding would suggest a need for further training on the management of MSK disease at the primary care level to avoid over-burdening the secondary care services.


Salt-assisted religation of proteolyzed Glutathione-S-transferase follows Hofmeister series.
Proteases have been used not only for proteolysis but also in organic solvent-assisted religation processes. Here, we demonstrated the effect of salts on peptide bond resynthesis in Glutathione-S-transferase (GST) and have found it to be in the purview of the Hofmeister phenomena. Our results show that the efficiency and ease of religation increases with an increase in the surface charge densities of the cations used in the study. Thus, the yield of religated GST follows the order: Mg2(+)>Li(+)>Na(+)>K(+). Characteristics of the salt-religated GST were studied using size exclusion chromatography, CD spectroscopy, mass spectrometry and CDNB activity assay. Results show that the properties of salt-religated GST are in close agreement with those of the native GST. Additionally, we also assessed the specific activity of the protease, Subtilisin Carlsberg, used in this study. Contrary, to aqueous-organic systems, wherein there is a remarkable decrease in the proteolytic activity, the activity in the presence of salts is only minimally changed. Our studies suggest that salt-assisted peptide bond formation is favoured primarily due to changes in the ionic environment of the nicked termini of GST, and that there is no role played by the protease.


Primary bacterial infection of the myocardium.
Primary bacterial infection of myocardial tissue without associated endocarditis occurs only rarely. It is generally seen in the setting of overwhelming bacteremia. The most common bacterial cause of myocarditis is Staphylococcus aureus, although infections with a broad range of bacterial pathogens have been described. Pathologically, the disease process is characterized by multifocal studding of the myocardium with tiny abscesses, and the left ventricle is most commonly involved. Complications include cardiac dysfunction, rhythm disturbances, and myocardial rupture with secondary purulent pericarditis. Since virtually all information regarding primary bacterial myocarditis originates from autopsy studies conducted in the pre-antibiotic era, little is known about the modern approach to diagnosis and management of this clinical entity.


Intubation of Profoundly Agitated Patients Treated with Prehospital Ketamine.
Profound agitation in the prehospital setting confers substantial risk to patients and providers. Optimal chemical sedation in this setting remains unclear. The goal of this study was to describe intubation rates among profoundly agitated patients treated with prehospital ketamine and to characterize clinically significant outcomes of a prehospital ketamine protocol. This was a retrospective cohort study of all patients who received prehospital ketamine, per a predefined protocol, for control of profound agitation and who subsequently were transported to an urban Level 1 trauma center from May 1, 2010 through August 31, 2013. Identified records were reviewed for basic ambulance run information, subject characteristics, ketamine dosing, and rate of intubation. Emergency Medical Services (EMS) ambulance run data were matched to hospital-based electronic medical records. Clinically significant outcomes are characterized, including unadjusted and adjusted rates of intubation. Overall, ketamine was administered 227 times in the prehospital setting with 135 cases meeting study criteria of use of ketamine for treatment of agitation. Endotracheal intubation was undertaken for 63% (85/135) of patients, including attempted prehospital intubation in four cases. Male gender and late night arrival were associated with intubation in univariate analyses (χ2=12.02; P=.001 and χ2=5.34; P=.021, respectively). Neither ketamine dose, co-administration of additional sedating medications, nor evidence of ethanol (ETOH) or sympathomimetic ingestion was associated with intubation. The association between intubation and both male gender and late night emergency department (ED) arrival persisted in multivariate analysis. Neither higher dose (>5mg/kg) ketamine nor co-administration of midazolam or haloperidol was associated with intubation in logistic regression modeling of the 120 subjects with weights recorded. Two deaths were observed. Post-hoc analysis of intubation rates suggested a high degree of provider-dependent variability. Prehospital ketamine is associated with a high rate of endotracheal intubation in profoundly agitated patients; however, ketamine dosing is not associated with intubation rate when adjusted for potential confounders. It is likely that factors not included in this analysis, including both provider comfort with post-ketamine patients and anticipated clinical course, play a role in the decision to intubate patients who receive prehospital ketamine. Olives TD , Nystrom PC , Cole JB , Dodd KW , Ho JD . Intubation of profoundly agitated patients treated with prehospital ketamine. Prehosp Disaster Med. 2016;31(6):593-602.