Panel-based next-generation sequencing identifies novel mutations in Bulgarian patients with inherited retinal dystrophies.
Next-generation sequencing (NGS)-based method is being used broadly for genetic testing especially for clinically and genetically heterogeneous disorders, such as inherited retinal degenerations (IRDs) but still not routinely used for molecular diagnostics in Bulgaria. Consequently, the purpose of this study was to evaluate the effectiveness of a molecular diagnostic approach, based on targeted NGS for the identification of the disease-causing mutations in 16 Bulgarian patients with different IRDs. We applied a customized NGS panel, including 125 genes associated with retinal and other eye diseases to the patients with hereditary retinopathies. Systematic filtering approach coupled with copy number variation analysis and segregation study lead to the identification of 16 pathogenic and likely pathogenic variants in 12/16 (75%) of IRD patients, 2 of which novel (12.5%): ABCA4-c.668delA (p.K223Rfs18) and RР1-c.2015dupA (p.K673Efs*25). Mutations in the ABCA4, PRPH2, USH2A, BEST1, RР1, CDHR1, and RHO genes were detected reaching a diagnostic yield between 42.9% for Retinitis pigmentosa cases and 100% for macular degeneration, Usher syndrome, and cone-rod dystrophy patients. Our results confirm the usefulness of targeted NGS approach based on frequently mutated genes as a comprehensive and successful genetic diagnostic tool for IRDs with significant impact on patients counseling.


Sleep disturbance as detected by actigraphy in pre-pubertal juvenile monkeys receiving therapeutic doses of fluoxetine.
Sleep disturbance is a reported side effect of antidepressant drugs in children. Using a nonhuman primate model of childhood selective serotonin reuptake inhibitor (SSRI) therapy, sleep was studied quantitatively with actigraphy. Two 48-h sessions were recorded in the home cage environment of juvenile male rhesus monkeys at two and three years of age, after one and two years of treatment with a therapeutic dose of the SSRI fluoxetine, and compared to vehicle treated controls. A third session was conducted one year after discontinuation of treatment at four years of age. During treatment, the fluoxetine group demonstrated sleep fragmentation as indexed by a greater number of rest-activity transitions compared to controls. In addition fluoxetine led to more inactivity during the day as indexed by longer duration of rest periods and the reduced activity during these periods. The fluoxetine effect on sleep fragmentation, but not on daytime rest, was modified by the monkey's genotype for polymorphisms of monoamine oxidase A (MAOA), an enzyme that metabolizes serotonin. After treatment, the fluoxetine effect on nighttime rest-activity transitions persisted, but daytime activity was not affected. The demonstration in this nonhuman primate model of sleep disturbance in connection with fluoxetine treatment and specific genetic polymorphisms, and in the absence of diagnosed psychopathology, can help inform use of this drug in children.


Differences in affinities of muscarinic acetylcholine receptor antagonists for brain and heart receptors.
The affinities of atropine, scopolamine, 3-quinuclidinyl benzilate and twelve analogues of 3-quinuclidinyl benzilate were determined for the muscarinic acetylcholine receptor (m-AChR) using membrane preparations from caudate/putamen. The affinity constants thus obtained were compared with affinities previously reported for the m-AChR obtained from ventricular muscle. The affinities differed significantly for six of the compounds, the largest difference being 16-fold. Neither solubilization nor variation of physiologically significant salts led to a significant change in the affinity of that compound. These results are interpreted as supporting the subclassification of the muscarinic acetylcholine receptor.


Drag force acting on a neuromast in the fish lateral line trunk canal. I. Numerical modelling of external-internal flow coupling.
Fishes use a complex, multi-branched, mechanoreceptive organ called the lateral line to detect the motion of water in their immediate surroundings. This study is concerned with a subset of that organ referred to as the lateral line trunk canal (LLTC). The LLTC consists of a long tube no more than a few millimetres in diameter embedded immediately under the skin of the fish on each side of its body. In most fishes, pore-like openings are regularly distributed along the LLTC, and a minute sensor enveloped in a gelatinous cupula, referred to as a neuromast, is located between each pair of pores. Drag forces resulting from fluid motions induced inside the LLTC by pressure fluctuations in the external flow stimulate the neuromasts. This study, Part I of a two-part sequence, investigates the motion-sensing characteristics of the LLTC and how it may be used by fishes to detect wakes. To this end, an idealized geometrical/dynamical situation is examined that retains the essential problem physics. A two-level numerical model is developed that couples the vortical flow outside the LLTC to the flow stimulating the neuromasts within it. First, using a Navier-Stokes solver, we calculate the unsteady flow past an elongated rectangular prism and a fish downstream of it, with both objects moving at the same speed. By construction, the prism generates a clean, periodic vortex street in its wake. Then, also using the Navier-Stokes solver, the pressure field associated with this external flow is used to calculate the unsteady flow inside the LLTC of the fish, which creates the drag forces acting on the neuromast cupula. Although idealized, this external-internal coupled flow model allows an investigation of the filtering properties and performance characteristics of the LLTC for a range of frequencies of biological interest. The results obtained here and in Part II show that the LLTC acts as a low-pass filter, preferentially damping high-frequency pressure gradient oscillations, and hence high-frequency accelerations, associated with the external flow.


One-month prophylactic use of OKT3 in cadaver kidney transplant recipients.
Fifty-five recipients of first cadaveric renal allografts were randomly assigned to three treatment groups in order to compare the safety and efficacy of a mouse antihuman T cell monoclonal antibody (OKT3) given prophylactically for a one-month period. This long period of administration was made possible by concomitant administration of azathioprine. The immune response against the foreign immunoglobulin was thus delayed and decreased in both intensity and severity, and OKT3 treatment could be given during almost the entire month in the majority of patients. The 18 patients who were enrolled in this treatment group had significantly (P less than 0.01) fewer rejection episodes during the first month posttransplantation than the 19 patients allocated to the high-dose (HD) steroid control group or the 18 patients allocated to the low-dose (LD) control group. Actual 2-year as well as actuarial 4-year graft survival rates were 89% in the OKT3 group, whereas they were 70% and 67% respectively in the steroid control groups. Four-year serum creatinine levels were normal and doses of steroids and other immunosuppressive agents were lower in the OKT3 group as compared with the control groups. Tolerance to OKT3 was good, and while viral infections were more frequently observed in OKT3 treated patients, the total number and the severity of infectious episodes were similar in all groups. The combination of one-month OKT3 plus azathioprine prophylaxis followed by conventional azathioprine plus low-dose steroid maintenance produced very satisfactory long-term results comparable to the best results now being obtained with cyclosporine regimens.


[Evolution of the profile of patients attended by a hospital Geriatric Home Care Unit in the last 20 years].
To describe the evolution of the profile of patients attended by a Geriatric Home Care (GHC) Unit and its care activity in the last two decades. Data on the historical activity of the AGD Unit from 2001 to 2020, grouped into 5-year periods, were recorded. Sociodemographic, clinical, functional (Functional Red Cross Scale and Barthel index) and mental (Mental Red Cross Scale) variables were collected, baseline and at inclusion to AGD. Also the waiting time until first visit, mean follow-up, origin of referral, destination at the end of the intervention, reason for consultation and cause of functional dependence were also included. A descriptive analysis was performed with the SPSSv.23 program. Ten thousand six hundred fifty-four patients attended in AGD (1 January 2001 to 31 December 2020). A progressive increase in age and in the number of geriatric syndromes was observed. Patients presented higher functional and cognitive decline, and the number of patients living alone and in need of private assistance increased. The duration of the intervention decreases and Primary Care remains the main source and destination at the end of the intervention, with dementia standing out as the main cause for functional dependence. The vulnerability of the populations in need of specialised geriatric care is increasing: patients are older, and have more geriatric syndromes, high functional and cognitive decline, and suffer social frailty; more work is needed to address the role of these support units in the community, as well as their coordination with Primary Care teams.


Preclinical assessment of anti-CD40 Mab 5D12 in cynomolgus monkeys.
Monoclonal antibody (Mab) 5D12 is a potent antagonist of the CD40-CD40L pathway. This cellular interaction has been validated in a large number of experimental animal models where dys-regulation of the immune system plays a role. Chimeric 5D12 (ch5D12) was constructed to reduce the potential immunogenicity and enhance the in vivo half-life when used in humans. ch5D12 is a molecularly engineered human IgG(4) antibody containing the variable domains of the heavy and light chains of the murine version of 5D12 (mu5D12). This new chimeric Mab was tested in a marmoset experimental autoimmune encephalomyelitis model and was shown to effectively prevent disease symptoms. The results of this in vivo evaluation supported clinical use of ch5D12 for immune targeted diseases. Therefore GMP material was prepared and a GLP-compliant tissue cross-reactivity study on human tissues (3 donors/37 tissues) and cynomolgus tissues (2 donors/37 tissues) was performed. ch5D12 stained on the surface of B cells and selected dendritic cells and no unexpected cross-reactivity was observed. The identical staining patterns in human and cynomolgus tissues justified the use of cynomolgus monkeys as a relevant model for humans. A GLP-compliant safety and tolerability evaluation for ch5D12 in cynomolgus monkeys was performed using the GMP produced material. Weekly administration of ch5D12 at two dose levels for 4 weeks was shown to be safe and without any side effects in all monkeys.


Scopolamine reverses haloperidol-attenuated lever-pressing for water but not haloperidol-attenuated water intake in the rat.
The operant lever-pressing response has previously (Ljungberg, Pharmacol Biochem Behav 27: 341-350, 1987) been found to be inhibited by lower doses of haloperidol than the corresponding consummatory act, i.e., water intake. In the present study it was found that the attenuation of the lever-pressing response caused by the neuroleptic, but not the attenuation of the water intake, could be counteracted by scopolamine. The results support the notion that blockade of operant responding by low doses of neuroleptics are probably related to the extra-pyramidal side-effects of neuroleptics seen in the clinic, as both phenomena can be counteracted by anticholinergics. These results therefore conflict with the anhedonia hypothesis put forward as an explanation of the attenuating effects of neuroleptics in operant settings. The findings also have a clear bearing on the role of dopamine in feeding and drinking behavior, as the results implies that different aspects of the control of water intake (i.e., the operant vs. the consummatory phase) are governed by different mechanisms in the CNS.


Molecular-Orbital Delocalization Enhances Charge Transfer in π-Conjugated Organic Semiconductors.
π-Conjugated organic semiconductors are promising materials for surface-enhanced Raman scattering (SERS)-active substrates based on the tunability of electronic structures and molecular orbitals. Herein, we investigate the effect of the temperature-mediated resonance-structure transitions of poly(3,4-ethylenedioxythiophene) (PEDOT) in poly(3,4-ethylenedioxythiophene)-poly(styrenesulfonate) (PEDOT : PSS) films on the interactions between substrate and probe molecules, thereby affecting the SERS activity. Absorption spectroscopy and density functional theory calculations show that this effect occurs mainly due to delocalization of the electron distribution in molecular orbitals, effectively promoting the charge transfer between the semiconductor and probe molecules. In this work, we investigate for the first time the effect of electron delocalization in molecular orbitals on SERS activity, which will provide new design ideas for the development of highly sensitive SERS substrates.


Relationship between plasma mid-regional pro-adrenomedullin level and resistance to antihypertensive therapy in stable kidney transplant recipients.
We investigated the relationship between plasma mid-regional pro-adrenomedullin (MR-proADM)-like immunoreactive substance (IS) level and clinical characteristics associated with renal failure or resistance to antihypertensive therapy in stable kidney transplant recipients. Forty-six Japanese kidney transplant recipients who underwent transplantation more than 90 days prior to the study were included. To evaluate resistance to antihypertensive therapy, we calculated the treatment intensity score of the antihypertensive drugs in each recipient. Morning blood samples were collected and plasma MR-proADM-IS levels were measured using an enzyme immunoassay. A significant correlation was observed between plasma MR-proADM-IS level with creatinine clearance or treatment intensity score. Multiple regression analysis identified plasma MR-proADM level and body mass index as significant independent factors associated with treatment intensity score. Plasma MR-proADM level may be a useful biomarker indicating the degree of resistance to antihypertensive therapy.


Stimulation of gastrointestinal motility by loperamide in dogs.
The effects of loperamide on gastrointestinal motility were investigated in conscious fasted dogs chronically fitted with strain-gauge transducers on the antrum, the jejunum, and the colon. Oral administration of loperamide (0.1 mg/kg) induced, after a delay of 20-30 min, a long-lasting (8-12 hr) stimulation of gastrointestinal motility associated with a disorganization of the cyclic activity at the three levels investigated. These effects were reproduced by a subcutaneous administration at the same dose and were antagonized by previous intravenous administration of naloxone or a quaternary opiate antagonist. Intracolonic administration (0.1 mg/kg) stimulated, after a delay of 20-30 min, colonic motility only. Intracerebroventricular loperamide (1 microgram/kg) induced a long-lasting (15-20 hr) inhibition of the gastric motility and a short (2-hr) disorganization of the jejunal motor profile. These data show that oral loperamide stimulates gastrointestinal motility in dogs and involves peripheral opiate receptors.


The process of wound healing of the avascular outer layers of the retina. Light- and electron microscopic studies on laser lesions of monkey eyes.
The process of wound healing of the avascular outer layers of the retina is studied on a series of Laser lesions of the monkey, Cercopithecus aethiops. The extent of the retinal lesions is limited to the pigment epithelium and photoreceptor cells, whereas Bruch's membrane and the vascular inner layers of the retina intact. The retinal lesions are not seen to be invaded by cells from the choroid, nor from the retinal vessels. During the first three days after irradiation the pigment epithelial cells in a zone, about 75 micrometer wide, around the lesions appear to be changed into bi- or multinucleate cells with the additional nuclei situated in apical cytoplasmic protrusions which extend towards and into the lesions. During the second and third days after irradiation the lesions are invaded by two kinds of cells which both appear to arise through the off-budding of the nucleus-containing apical protrusions of the pigment epithelial cells around the lesions: (a) phagocytic cells which disintegrate the damaged tissue and, as this process is completed 4--9 days after irradiation show evidence of melanogenesis; and (b) regenerated pigment epithelium which form a single layer of cells along Bruch's membrane, but show no evidence of phagocytosis or intracellular disintegration of the damaged tissue. Within the lesions both these kinds of cells show mitotic figures the third day after irradiation.


Blood to the cornual area of the uterus is mainly supplied from the ovarian artery in the follicular phase and from the uterine artery in the luteal phase.
The blood supply to the uterus is provided by the uterine and ovarian arteries, which form anastomoses. Yet the flow direction through this anastomoses and the primary source of blood supply to the tubes and uterine cornua remains unknown. To clarify this issue, we studied the spatial propagation of temperature changes following cooling of the upper vaginal area. A thermocatheter with eight measurement points at 5-mm intervals was inserted into the uterus of nine women in the follicular phase and 11 in the luteal phase. The distal tip was positioned in the cornual area and temperatures were registered every 2 s. The vagina was then cooled for 7 min with 25 degrees C saline. The pattern of uterine cooling based on local counter-current transfer differed between the follicular and luteal phase. Cooling of the cornual area was significantly lower in the luteal phase compared with the follicular phase, indicating a shift in the prevailing source of arterial supply in that area following ovulation. The divide between the territories irrigated by the uterine and ovarian arteries moves between the follicular and luteal phase. This constitutes the first description of a functionally determined shift in the territorial divide of two vascular systems, and has numerous practical implications.


Correlation of presence of granulomas with clinical and immunologic variables in Crohn's disease.
Approximately 50% of patients with Crohn's disease have epithelioid granulomas present in the diseased intestine. Some studies have associated the presence of granulomas with a good prognosis. In this prospective study, 44 patients with Crohn's disease requiring surgery were followed up for five years. Twenty-two patients (50%) had granulomas. Patients with granulomas were younger and had a shorter duration of disease. They also had more extensive disease and a greater degree of peripheral lymphopenia. Follow-up showed a trend toward greater recurrence rate in the patients with granulomas. It seems that patients with aggressive and extensive Crohn's disease are not protected from the development of symptomatic early recurrence by the presence of epithelioid granulomas.


Nearby grandmother enhances calf survival and reproduction in Asian elephants.
Usually animals reproduce into old age, but a few species such as humans and killer whales can live decades after their last reproduction. The grandmother hypothesis proposes that such life-history evolved through older females switching to invest in their existing (grand)offspring, thereby increasing their inclusive fitness and selection for post-reproductive lifespan. However, positive grandmother effects are also found in non-menopausal taxa, but evidence of their associated fitness effects is rare and only a few tests of the hypothesis in such species exist. Here we investigate the grandmother effects in Asian elephants. Using a multigenerational demographic dataset on semi-captive elephants in Myanmar, we found that grandcalves from young mothers (<20 years) had 8 times lower mortality risk if the grandmother resided with her grandcalf compared to grandmothers residing elsewhere. Resident grandmothers also decreased their daughters' inter-birth intervals by one year. In contrast to the hypothesis predictions, the grandmother's own reproductive status did not modify such grandmother benefits. That elephant grandmothers increased their inclusive fitness by enhancing their daughter's reproductive rate and success irrespective of their own reproductive status suggests that fitness-enhancing grandmaternal effects are widespread, and challenge the view that grandmother effects alone select for menopause coupled with long post-reproductive lifespan.


Rural non-farm income diversification: implications on smallholder farmers' welfare and agricultural technology adoption in Ghana.
This study explored the potential impact of rural non-farm income diversification on households' welfare and adoption of Zai-technology (a proxy for agricultural technology adoption) using primary data collected from agricultural households in the Upper East region of Ghana. We used Propensity Score Matching (PSM) and Inverse-Probability-weighted Regression Adjustment (IPWRA) techniques to estimate welfare and Zai-technology impacts of non-farm income diversification. After controlling for differences in covariates, the results show that non-farm income diversification increases the likelihood of Zai-technology adoption and contributes to significant household welfare gains. We therefore suggest that the activities of agricultural extension services and farmer-based organizations (FBOs) be enhanced as they facilitate the diversification of non-farm incomes, thereby increasing investment in productivity-enhancing technologies (Zai) and household welfare.


Comparison of bicyclic phosphorous esters with bicuculline and picrotoxin as antagonists of presynaptic inhibition in the rat cuneate nucleus.
The effects of 2 of a series of bicyclic phosphorous esters, the ethyl (EPTBO) and isopropyl (IPTBO) compounds, were compared with those of the GABA antagonists, picrotoxin and bicuculline, on presynaptic inhibition in the rat cunneate nucleus. Both EPTBO and IPTBO were found to be effective, reversible antagonists of presynaptic inhibition, with IPTBO approximately 10 times more potent than EPTBO and equipotent with bicuculline, EPTBO equipotent with picrotoxin.


Combined immunostaining of neurofilaments, neuron specific enolase, GFAP and S-100. A possible means for assessing the morphological and functional status of the enteric nervous system.
Neurofilaments, part of the cytoskeletal network, and neuron specific enolase, a major enzyme in glycolysis, are both present in central and peripheral neurons. Glial fibrillary acidic protein and S-100, on the other hand, are soluble proteins which are found exclusively in the supportive cells of the nervous system, i.e. the glial cells. Examination was made, using immunocytochemistry, of all main areas of the gastrointestinal tract of three mammalian species, rat, pig and man. By applying serial tissue sectioning, it was possible to study the relative occurrences of the two neuronal markers in the same cell bodies and to examine the relationships of the neurons with the glial cells as revealed by the antibodies to glial fibrillary acidic protein and S-100. Both neurofilaments and neuron specific enolase were localised to an extensive system of enteric nerves, with the level of neuron specific enolase-immunoreactivity showing greater variability than that observed using antibodies to neurofilaments. Comparison of the occurrence of neuron specific enolase and neurofilament immunoreactivity in serially sectioned neuronal cell bodies revealed that a minor population stained only with antibodies to neurofilaments. The equivocal or absent neuron specific enolase-immunoreactivity in some perikarya may reflect variations in functional status within the nervous system. Glial fibrillary acidic protein- and S-100-immunoreactivities were confined to glial cells which, in this normal tissue, were always in close association with the neurons.(ABSTRACT TRUNCATED AT 250 WORDS)


Environmental impact assessment of dam-break floods considering multiple influencing factors.
Dam-break floods cause substantial damage to the environment, and evaluating the negative impacts of dam-break floods on the environment (EI) is an important part of flood risk management. EI has been evaluated using various methods with different indices. However, the evaluation results of EI are typically one-sided or inaccurate because of diverse indices and complex influencing factors. A new method was proposed herein to calculate EI with an index system, including geomorphic changes (GC), water pollution (WP), plant biomass loss (PB), and biodiversity loss (BL). Eight factors that influence EI were sorted out, namely, erosion or deposition depth, non-point source (NPS) and point source (PS) pollution, plant biomass, species richness, plant height, and flood depth, velocity, and duration. After combining the proposed damage functions to calculate the influence of flood depth, velocity, and duration, and plant height on the environment, methods to calculate GC, WP, PB, BL, and EI were proposed. A dam-break flood scenario for Luhun Reservoir was used to verify the method. The results showed that (1) the trend in EI was similar to that in geomorphic changes in the inundated area and seriously affected by PS in local areas, (2) the average EI of woodland was the highest, while that of towns was the slowest, and (3) GC and WP contributed 93.7% of EI in the entire inundated area. This study summarized the complex impacts of dam-break floods on the environment from four aspects and proposed a method to quantify the overall impact of dam-break floods on the environment. The evaluation model could evaluate the impact of floods on the environment accurately, presenting the results on a flood inundation map. This provides a scientific basis for evaluating flood consequences and managing flood risk.


[The influence of some carbamazepin metabolites on Nielsen's quantitative determination. 5th communication: on chemistry and analytics of dibenzazepine derivatives (author's transl)].
H. R. Nielsen and H. Remmer described a quantitative method for the estimation of carbamazepin serum levels. The specificity of this method was examined with the result that besides carbamazepin also its metabolites react under the authors' conditions.


[Comparison of algorithms for the resolution of a secular equation in the vibrational analysis of biological molecules].
A computer analysis of several solution methods for secular equations leads to a determination of their optimum conditions. The results contribute to the application of a new preliminary method of frequency assignment to the vibrational study of the heavy atoms coordination sphere in complex biological structures, such as metalporphyrins and adenosine triphosphate.


Supporting biomedical ontology evolution by identifying outdated concepts and the required type of change.
The consistent evolution of ontologies is a major challenge for systems using semantically enriched data, for example, for annotating, indexing, or reasoning. The biomedical domain is a typical example where ontologies, expressed with different formalisms, have been used for a long time and whose dynamic nature requires the regular revision of underlying systems. However, the automatic identification of outdated concepts and proposition of revision actions to update them are still open research questions. Solutions to these problems are of great interest to organizations that manage huge and dynamic ontologies. In this paper, we present an approach for (i) identifying the concepts of an ontology that require revision and (ii) suggesting the type of revision. Our analysis is based on three aspects: structural information encoded in the ontology, relational information gained from external source of knowledge (i.e., PubMed and UMLS) and temporal information derived from the history of the ontology. Our approach aims to evaluate different methods and parameters used by supervised learning classifiers to identify both the set of concepts that need revision, and the type of revision. We applied our approach to four well-known biomedical ontologies/terminologies (ICD-9-CM, MeSH, NCIt and SNOMED CT) and compared our results to similar approaches. Our model shows accuracy ranging from 68% (for SNOMED CT) to 91% (for MeSH), and an average of 71% when considering all datasets together.


Adverse effects of carbamazepine monotherapy among patients in Nigeria: a pilot study and implications.
Antiepileptic drugs do cause adverse effects, affecting patients' quality of life, adherence and seizures. Consequently, there is a need to study this among Nigerian patients. Descriptive cross-sectional study assessing the extent of adverse effects with carbamazepine monotherapy and potential factors. 54 patients (64.3%) reported no adverse effects, while 30 did. The most common adverse effects were memory problems followed by headaches, restlessness, tiredness and depression. Adverse effects were significantly higher in females, with females with low educational levels having significantly higher rates. Dose, co-medications, seizure control and comorbid conditions did not significantly affect adverse effects. Perception of adverse effects was common in patients on carbamazepine monotherapy, more common in women than men. Educational status was important in women.


Gonadotropin-releasing hormone agonist for the preservation of ovarian function in survivors of haematopoietic stem cell transplantation for haematological diseases.
Administration of GnRH agonist (GnRHa) prior to chemotherapy may decreases the risk of gonadal dysfunction in patients with tumors. However, relevant data in haematopoietic stem cell transplantation (HSCT) recipients has not yet been established. Hence, the present study was designed to evaluate the clinical efficacy of GnRHa cotreatment prior to myeloablative regimens on ovarian protection in female survivors of HSCT for haematological diseases. Eligible patients were divided into a GnRHa group and a control group. Medical records regarding age at HSCT; diagnosis/indication for HSCT; pre- and posttransplantation serum sex hormone levels; menstruation and perimenopausal symptoms after HSCT were collected and compared. The primary and secondary outcome was the incidence of premature ovarian insufficiency (POI) symptoms associated with hypoestrogenism. A total of 330 patients were enrolled in the study: 19 patients were lost to follow-up, and clinical information was obtained in 311 patients. There was no significant difference in the primary outcome of follow-up between the two groups (78.50% [84 of 107] for the GnRHa group versus 83.33% [170 of 204] for the control group). The adjusted relative risks (RR) and 95% confidence interval (CI) were 1.19 and 0.73-1.93 (P = 0.487). Among patients who received cotreatment with GnRHa, 62.62% (67 of 107) complained of perimenopausal symptoms, which was significantly lower than the 74.51% (152 of 204) in the control group (adjusted RR: 1.46, 95% CI: 1.04-2.06, P = 0.031). GnRHa cotreatment may not decrease the POI rate in HSCT survivors. However, it may reduce perimenopausal symptoms in this population, suggesting a potential benefit of GnRHa in clinical practice and warrant further researches.


A Crisis for a System in Crisis: Forecasting from the Short- and Long-Term Impacts of COVID-19 on the Child Welfare System.
The COVID-19 pandemic has thrust the world into a crisis - and the child welfare system is particularly susceptible to its effects. This pandemic has exacerbated some of the most problematic aspects of the system, and its impacts will reverberate long after the immediate crisis ends. As COVID-19 spread, families were instantly impacted - in-person family time was cancelled, youth and families were unable to access basic resources, services, and technology, and access to the courts was curtailed. Those short-term effects may give way to long-term harms such as disrupted attachments and delays in achieving permanency. The pandemic also reinforced the importance of key tenets of a well-functioning child welfare system: high-quality legal representation, creativity, and youth and family engagement. Attorneys must learn from the fallout of the pandemic, retain the best responsive practices, and use the lessons learned from this crisis to transform dependency cases, and the system writ large, into what families need and deserve.


β-Carotene biofortification of chia sprouts with plant growth regulators.
Chia (Salvia hispanica) is a native plant species from South America that is very appreciated for its oleaginous seeds in the agri-food field. Chia seeds are natural sources of many bioactive compounds which provide benefits to human health. Nevertheless, chia sprouts have better nutritional properties than seeds, such as antioxidants, essential amino acids, and phenolic compounds. Among all these beneficial compounds, β-carotene has not been studied in chia sprouts. β-carotene is a precursor of vitamin A, which contributes to maintaining our health status. In this study, to improve β-carotene content in chia sprouts, some plant growth regulators (abscisic acid, methyl jasmonate and methyl salicylate) were applied exogenously to germinating chia seeds. Gibberellins A4/A7 and cytokinin 6-benzyladenine (Promalin®) were also applied, combined with the other regulators, to antagonize a possible inhibition in the germination. Seeds were grown in darkness for 4 days, then seeds were exposed to a short light stimulus (30') and finally to a continued light stimulus (48h). β-carotene, xanthophylls, chlorophylls, de-epoxidation status of xanthophyll cycle (DPS), germination rate, and sprouts fresh weight were analysed. The results show that sprouts treated with methyl salicylate in-creased 2,35 fold their β-carotene content when they were exposed to light for 30'+48h. Sprouts fresh weight and germination were not affected by methyl salicylate. Although more research is needed before industrial application, it is concluded that methyl salicylate can be used to improve β-carotene contents in chia sprouts.


A label-free fluorescent molecular beacon based on DNA-templated silver nanoclusters for detection of adenosine and adenosine deaminase.
A simple and reliable fluorescent molecular beacon is developed utilizing DNA-templated silver nanoclusters as a signal indicator and adenosine triphosphate (ATP) and adenosine deaminase as mechanical activators.


Cortisol metabolism in hypertensive patients who do and do not develop hypokalemia from diuretics.
Deficiency of 11 beta-hydroxysteroid dehydrogenase causes hypertension and hypokalemia. To test whether hypertensive patients who develop hypokalemia when treated with diuretics have low levels of activity of this enzyme as a metabolic predisposition to the development of hypokalemia, we measured urinary cortisone, cortisol, tetrahydrocortisol, tetrahydrocortisone, and creatinine in 42 hypertensive patients who either did or did not become hypokalemic on hydrochlorothiazide. The mean ratios of cortisone to cortisol, tetrahydrocortisone to tetrahydrocortisol, tetrahydrocortisol to cortisol, and cortisol to creatinine did not differ between the two groups. We conclude that hypertensives who develop hypokalemia on diuretics do not have low activity of this enzyme. They also do not appear to have low ring A reduction or higher cortisol secretion rates compared with hypertensives who do not develop hypokalemia. We failed to find a metabolic predisposition to the development of hypokalemia by diuretic treatment.


Erdheim-Chester disease of the central nervous system: new manifestations of a rare disease.
ECD is a rare non-Langerhans-cell histiocytosis, which can involve the CNS; therefore, CNS imaging findings have been described in only a small number of patients. To gain additional insight into the CNS manifestations of ECD, we reviewed the findings on imaging of the brain, head and neck, and spine in patients with ECD who presented to our institution. Here, we illustrate manifestations that have not, to our knowledge, been previously described. CT, MR imaging, and PET/CT studies of the brain, maxillofacial region, and spine were reviewed in 11 patients with ECD. Four new manifestations of ECD were present, including the following: a stellate appearance of intracranial extra-axial lesions, ependymal enhancement along the lateral ventricle with deep linear extension to the lentiform nucleus, irregular enhancement in the pons, and diffuse involvement of the vertebral column on PET/CT. ECD has a variety of imaging appearances in the CNS, including new manifestations described herein. Neuroradiologists should be aware of these manifestations to avoid mistaking them for other disease processes.


Indomethacin in the treatment of idiopathic minimal lesion nephrotic syndrome.
Ten patients with idiopathic minimal lesion nephrotic syndrome were treated with indomethacin. Nine patients received the drug for five weeks and one patient for nine weeks. Indomethacin was administered in a dose of 1 mg/kg/day during the first week and 2 mg/kg/day, thereafter. At the end of the study, nine patients remained edematous and all had a urinary protein excretion greater than 50 mg/kg/day, within the nephrotic range. Indomethacin did not change the course of the disease and its use in the treatment of IMLNS is not advised.


Interaction between primary alveolar macrophages and primary alveolar type II cells under basal conditions and after lipopolysaccharide or quartz exposure.
Intercellular communications between alveolar macrophages (AM) and alveolar epithelial type II (TII) cells have been suggested to be important in cellular responses. The main objective of this study was to improve our understanding of the interactions between AM and TII cells that might occur in the lung. In the present investigation, this interaction was studied under different interaction conditions (transwell or mixed coculture) and different exposure conditions (basal, lipopolysaccharide [LPS] exposure, or silica exposure). Studies also attempted different approaches to identify specific mediator(s) involved in this interaction. The following findings were made: (1) Surfactant released from TII cells appears to exert an inhibitory effect on AM activity. (2) Basal transwell coculture conditions are better than mixed coculture conditions to study AM/TII cell interactions, since the inhibitory effect of the surfactant in the transwell coculture is minimized. (3) AM/TII cell interaction is dependent on cell culture (transwell vs mixed) and exposure conditions. (4) Oxidants, tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta, prostaglandins, and leukotrienes probably do not independently affect the AM/TII intercellular interaction; instead, they appear to indirectly modulate the complex pathways of AM/TII communication.


Posttransplant hematopoiesis in patients undergoing sibling allogeneic stem cell transplantation reflects that of their respective donors although with a lower functional capability.
We tested the principle of whether patient long-term hematopoiesis following allogeneic stem cell transplantation (allo-SCT) reflects the characteristics of the hematopoiesis of their respective donor. For this purpose, we analyzed bone marrow (BM) hematopoiesis using long-term cultures (LTC), delta assays, and clonogeneic assays as well as CD34+ cells and their subsets by flow cytometry in a series of 37 patients undergoing allo-SCT, and we compared it to that of their respective human leukocyte antigen-matched sibling donors in a paired study performed more than 1 year after the transplant procedure. Interestingly, the main factor that influenced post-allo-SCT BM hematopoiesis in the long term was donor hematopoiesis. Nevertheless, compared to their respective donors, patients exhibited a significantly lower number of colony-forming units granulomonocytic, burst-forming units erythroid, and immature progenitors (CD34++/CD38dim/CD90+/CD133+ cells, LTC-initiating cells, and colonies generated in the delta assay). Moreover, BM stromal function was diminished in patients undergoing allo-SCT compared to their donors. In addition, the presence of chronic graft-versus-host disease under immunosuppressive treatment also conditioned an impaired hematopoietic function. In summary, our study shows that BM hematopoiesis evaluated more than 1 year after an allo-SCT mainly reproduces that of their respective donors, although with a significantly decreased in vitro activity.


Prescribing "placebo treatments": results of national survey of US internists and rheumatologists.
To describe the attitudes and behaviours regarding placebo treatments, defined as a treatment whose benefits derive from positive patient expectations and not from the physiological mechanism of the treatment itself. Cross sectional mailed survey. Physicians' clinical practices. 1200 practising internists and rheumatologists in the United States. Investigators measured physicians' self reported behaviours and attitudes concerning the use of placebo treatments, including measures of whether they would use or had recommended a "placebo treatment," their ethical judgments about the practice, what they recommended as placebo treatments, and how they typically communicate with patients about the practice. 679 physicians (57%) responded to the survey. About half of the surveyed internists and rheumatologists reported prescribing placebo treatments on a regular basis (46-58%, depending on how the question was phrased). Most physicians (399, 62%) believed the practice to be ethically permissible. Few reported using saline (18, 3%) or sugar pills (12, 2%) as placebo treatments, while large proportions reported using over the counter analgesics (267, 41%) and vitamins (243, 38%) as placebo treatments within the past year. A small but notable proportion of physicians reported using antibiotics (86, 13%) and sedatives (86, 13%) as placebo treatments during the same period. Furthermore, physicians who use placebo treatments most commonly describe them to patients as a potentially beneficial medicine or treatment not typically used for their condition (241, 68%); only rarely do they explicitly describe them as placebos (18, 5%). Prescribing placebo treatments seems to be common and is viewed as ethically permissible among the surveyed US internists and rheumatologists. Vitamins and over the counter analgesics are the most commonly used treatments. Physicians might not be fully transparent with their patients about the use of placebos and might have mixed motivations for recommending such treatments.


Acute phenylalanine/tyrosine depletion of phasic dopamine in the rat brain.
Dopamine plays a critical role in striatal and cortical function, and depletion of the dopamine precursors phenylalanine and tyrosine is used in humans to temporarily reduce dopamine and probe the role of dopamine in behavior. This method has been shown to alter addiction-related behaviors and cognitive functioning presumably by reducing dopamine transmission, but it is unclear what specific aspects of dopamine transmission are altered. We performed this study to confirm that administration of an amino acid mixture omitting phenylalanine and tyrosine (Phe/Tyr[-]) reduces tyrosine tissue content in the prefrontal cortex (PFC) and nucleus accumbens (NAc), and to test the hypothesis that Phe/Tyr[-] administration reduces phasic dopamine release in the NAc. Rats were injected with a Phe/Tyr[-] amino acid mixture, a control amino acid mixture, or saline. High-performance liquid chromatography was used to determine the concentration of tyrosine, dopamine, or norepinephrine in tissue punches from the PFC and ventral striatum. In a separate group of rats, phasic dopamine release was measured with fast-scan cyclic voltammetry in the NAc core after injection with either the Phe/Tyr[-] mixture or the control amino acid solution. Phe/Tyr[-] reduced tyrosine content in the PFC and NAc, but dopamine and norepinephrine tissue content were not reduced. Moreover, Phe/Tyr[-] decreased the frequency of dopamine transients, but not their amplitude, in freely moving rats. These results indicate that depletion of tyrosine via Phe/Tyr[-] decreases phasic dopamine transmission, providing insight into the mechanism by which this method modifies dopamine-dependent behaviors in human imaging studies.


Otoscopy and tympanometry outcomes from the National Health and Nutrition Examination Survey (NHANES).
The purpose of the study was to estimate the prevalence of conductive pathologies based on tympanometric and otoscopic findings in adults in the U.S. and examine relationships between abnormal findings and audiometrically defined or reported perceived hearing loss and tinnitus. Data from 3409 individuals aged 20-69 years from the 1999-2000 and 2000-2002 cycles of the National Health and Nutrition Examination Survey (NHANES) were analyzed. Outcomes included report of ear tubes, self-perceived hearing loss, and tinnitus (yes or no). Otoscopy involved screening for presence of cerumen (ear wax) and physical abnormality for each ear. Ear-specific tympanometry included measures of middle ear pressure, external ear volume, tympanometric width, and admittance/compliance for each ear. Mean and prevalence estimates are provided applying sample weights. A multivariate ordinal regression model adjusting for age, sex, race/ethnicity, and education was used to examine relationships between otoscopy or tympanometry measures with hearing loss and tinnitus. Approximately 12% of the sample showed evidence of abnormal tympanometry and 12% showed evidence of abnormal otoscopy in at least one ear. Measured hearing loss was consistently related to tympanometry outcomes, in both univariate and adjusted models, but perceived hearing loss and tinnitus were not related to abnormal tympanometry or otoscopy. The most common reason for abnormal otoscopy was presence of excessive or impacted cerumen, representing an estimated 10% of the population. With 1 out of 10 adults having excessive cerumen, adults considering over-the-counter (OTC) hearings aids may benefit from an ear examination prior to purchase.


Challenges to Telehealth: What Was Learned from Families of Children with Neurodevelopmental Disorders.
Children with neurodevelopmental disorders (NDD) are a vulnerable population diagnosed as having an impairment of the central nervous system caused by genetic, metabolic, toxic or traumatic factors. During the coronavirus disease 2019 (COVID-19) pandemic children with NDD benefitted from the swift transition to telehealth. Most found telehealth favorable but some encountered challenges with accessibility and technology. Racial disparity was found with accessibility challenges by marginalized groups within this already vulnerable population. Telehealth use should continue to be utilized by children with NDD but clinicians should be aware of how to address challenges.


Portable Prehospital Methods to Treat Near-Hypothermic Shivering Cold Casualties.
To compare the effectiveness of a single-layered polyethylene survival bag (P), a single-layered polyethylene survival bag with a hot drink (P+HD), a multi-layered metalized plastic sheeting survival bag (MPS: Blizzard Survival), and a multi-layered MPS survival bag with 4 large chemical heat pads (MPS+HP: Blizzard Heat) to treat cold casualties. Portable cold casualty treatment methods were compared by examining core and skin temperature, metabolic heat production, and thermal comfort during a 3-hour, 0°C cold air exposure in 7 shivering, near-hypothermic men (35.4°C). The hot drink (70°C, ~400 ml, ~28 kJ) was consumed at 0, 1, and 2 hours during the cold air exposure. During the cold air exposure, core rewarming and thermal comfort were similar on all trials (P = .45 and P = .36, respectively). However, skin temperature was higher (10%-13%; P < .001; large effect sizes d > 2.7) and metabolic heat production lower (15%-39%; P < .05; large effect sizes d > .9) on MPS and MPS+HP than P and P+HD. The addition of heat pads further lowered metabolic heat production by 15% (MPS+HP vs MPS; P = .05; large effect size d = .9). The addition of the hot drink to polyethylene survival bag did not increase skin temperature or lower metabolic heat production. Near-hypothermic cold casualties are rewarmed with less peripheral cold stress and shivering thermogenesis using a multi-layered MPS survival bag compared with a polyethylene survival bag. Prehospital rewarming is further aided by large chemical heat pads but not by hot drinks.


Comparison of usefulness of the wall thickness of the left anterior descending coronary artery, determined by transthoracic echocardiography, and carotid intima-media thickness in predicting multivessel coronary artery disease.
Wall thickness in the distal part of the left anterior descending coronary artery (LAD) can be measured by using two-dimensional high-resolution transthoracic echocardiography (2DHTTE). The objective of this study was to compare the diagnostic accuracy of measuring carotid intima-media thickness (IMT) and distal LAD wall thickness for prediction of multivessel coronary artery disease (MVD). We measured wall thickness in the distal LAD using 2DHTTE and carotid IMT using B-mode ultrasound in 100 patients who subsequently underwent coronary angiography (CAG). Patients were classified into three groups based on the results of CAG-no significant stenosis (group N), single-vessel disease (group S), or multivessel disease (group M). Successful measurements of LAD wall thickness were accomplished in 96 patients. Distal LAD wall thickness was significantly greater in group M (0.92 ± 0.20 mm) than in group N (0.72 ± 0.11 mm, P < 0.01) and group S (0.76 ± 0.19 mm, P < 0.01). Carotid IMT was significantly greater in group M (0.90 ± 0.24 mm) than in group N (0.75 ± 0.21 mm, P < 0.05) and group S (0.80 ± 0.17 mm, P < 0.05). Distal LAD wall thickness >0.8 mm had a sensitivity of 75% and a specificity of 67% in predicting MVD, whereas carotid IMT >0.8 mm was 63% sensitive and 67% specific in the prediction of MVD. There was a weak but significant correlation between distal LAD wall thickness and IMT (r = 0.31, P < 0.01). Non-invasive measurement of distal LAD wall thickness by 2DHTTE is feasible, and has equivalent diagnostic accuracy to IMT measurements for predicting MVD.


Antisocial behavioral syndromes and 3-year quality-of-life outcomes in United States adults.
To examine 3-year quality-of-life (QOL) outcomes among United States adults with Diagnostic and Statistical Manual of Mental Disorders - Fourth Edition (DSM-IV) antisocial personality disorder (ASPD), syndromal adult antisocial behavior without conduct disorder (CD) before age 15 [adulthood antisocial behavioral syndrome (AABS), not a DSM-IV diagnosis], or no antisocial behavioral syndrome at baseline. Face-to-face interviews (n = 34 653). Psychiatric disorders were assessed using the Alcohol Use Disorder and Associated Disabilities Interview Schedule - DSM-IV Version. Health-related QOL was assessed using the Short-Form 12-Item Health Survey, version 2 (SF-12v2). Other outcomes included past-year Perceived Stress Scale-4 (PSS-4) scores, employment, receipt of Supplemental Security Income (SSI), welfare, and food stamps, and participation in social relationships. Antisocial personality disorder and AABS predicted poorer employment, financial dependency, social relationship, and physical health outcomes. Relationships of antisociality to SSI and food stamp receipt and physical health scales were modified by baseline age. Both antisocial syndromes predicted higher PSS-4, AABS predicted lower SF-12v2 Vitality, and ASPD predicted lower SF-12v2 Social Functioning scores in women. Similar prediction of QOL by ASPD and AABS suggests limited utility of requiring CD before age 15 to diagnose ASPD. Findings underscore the need to improve prevention and treatment of antisocial syndromes.


Red ink tattoo reactions: successful treatment with the Q-switched 532 nm Nd:YAG laser.
In the South-west Thames region there were an unprecedented number of lichenoid tattoo reactions to red ink in patients who had visited a local tattoo parlour. The red ink was found to contain mercuric sulphide, a compound known to cause allergic reactions. Topical Dermovate (clobetasol propionate 0.05%, GlaxoWellcome) ointment alone had little impact. To investigate whether the Q-switched 532 nm Nd:YAG laser could produce permanent flattening of the reaction. This was an open nonrandomized clinical trial. Biopsies were taken from the lichenoid areas within the tattoos. Subjects were patch tested to 1% ammoniated mercury in petrolatum prior to treatment with the Q-switched 532 nm Nd:YAG laser. Laser treatments were delivered at 6-weekly intervals by a single operator. Patients also applied topical Dermovate between treatments. Therapy was discontinued when the lesions flattened. Clinical photographs were assessed at baseline and prior to each laser treatment. Seven patients with Fitzpatrick skin types I-III were enrolled in the study (four females, three males, mean age 39 years). All patients completed the trial. Patch testing to mercury was universally negative at 48 and 96 h. Substantial flattening and depigmentation of the red ink within the tattoos was noted after six laser treatments. No adverse effects were recorded. The Q-switched 532 nm Nd:YAG laser in combination with topical Dermovate ointment is a safe and effective method of treating red ink tattoo reactions.


Thermally controlled intein splicing of engineered DNA polymerases provides a robust and generalizable solution for accurate and sensitive molecular diagnostics.
DNA polymerases are essential for nucleic acid synthesis, cloning, sequencing and molecular diagnostics technologies. Conditional intein splicing is a powerful tool for controlling enzyme reactions. We have engineered a thermal switch into thermostable DNA polymerases from two structurally distinct polymerase families by inserting a thermally activated intein domain into a surface loop that is integral to the polymerase active site, thereby blocking DNA or RNA template access. The fusion proteins are inactive, but retain their structures, such that the intein excises during a heat pulse delivered at 70-80°C to generate spliced, active polymerases. This straightforward thermal activation step provides a highly effective, one-component 'hot-start' control of PCR reactions that enables accurate target amplification by minimizing unwanted by-products generated by off-target reactions. In one engineered enzyme, derived from Thermus aquaticus DNA polymerase, both DNA polymerase and reverse transcriptase activities are controlled by the intein, enabling single-reagent amplification of DNA and RNA under hot-start conditions. This engineered polymerase provides high-sensitivity detection for molecular diagnostics applications, amplifying 5-6 copies of the tested DNA and RNA targets with >95% certainty. The design principles used to engineer the inteins can be readily applied to construct other conditionally activated nucleic acid processing enzymes.


Emotional eating in overweight, normal weight, and underweight individuals.
Emotional states and situations can affect food intake. We predicted that underweight individuals would eat less and overweight individuals would eat more during negative as well as positive emotional states and situations. Questionnaires to assess eating during emotional states and situations were distributed and collected in person in several major university and public libraries. Ninety questionnaires, representing for each gender the 15 most overweight, the 15 closest to normal weight, and the 15 most underweight, were analyzed. Gender had only minor effects on the eating ratings, and therefore the results are presented for the sexes combined. Underweight individuals reported eating less (P=.000) than both the normal and overweight groups during negative emotional states and situations. More surprisingly, underweight individuals also reported eating more (P=.01) than the other groups during positive emotional states and situations. Thus, part of the prediction was confirmed: the relative undereating by the underweight group, and the relative overeating by the overweight group during negative emotional states and situations. As compared to their usual eating behavior, undereating by underweight individuals during negative emotional states and situations was of a greater magnitude than their own overeating during positive states and situations (P=.01). Undereating by underweight individuals when experiencing negative emotions may contribute to their low body weight.


Quantitative determination of phosphatidylcholine hydroperoxides during copper oxidation of LDL and HDL by liquid chromatography/mass spectrometry.
1-Palmitoyl-2-linoleoylphosphatidylcholine monohydroperoxide (PC 16:0/18:2-OOH) and 1-stearoyl-2-linoleoylphosphatidylcholine monohydroperoxide (PC 18:0/18:2-OOH) were measured by liquid chromatography/mass spectrometry (LC/MS) using nonendogenous 1-palmitoyl-2-heptadecenoylphosphatidylcholine monohydroperoxide as an internal standard. The calibration curves for synthetic PC 16:0/18:2-OOH and PC 18:0/18:2-OOH, which were obtained by direct injection of the internal standard into the LC/MS system, were linear throughout the calibration range (0.8-12.8 pmol). Within-day and between-day coefficients of variation were less than 10%, and the recoveries were between 86% and 105%. The limit of detection (LOD) and the limit of quantification (LOQ) were determined using synthetic standards. The LOD (signal-to-noise ratio 3:1) was 0.01 pmol, and the LOQ (signal-to-noise ratio 6:1) was 0.08 pmol for both PC 16:0/18:2-OOH and PC 18:0/18:2-OOH. With use of this method, the concentrations of PC 16:0/18:2-OOH and PC 18:0/18:2-OOH in the lipoprotein fractions during copper-mediated oxidation were determined. We prepared oxLDL and oxHDL by incubating native LDL and native HDL from human plasma (n = 10) with CuSO(4) for up to 4 h. The time course of the PC 16:0/18:2-OOH and PC 18:0/18:2-OOH levels during oxidation consisted of three phases. For oxidized LDL, both compounds exhibited a slow lag phase and a subsequent rapidly increasing propagation phase, followed by a gradually decreasing degradation phase. In contrast, for oxidized HDL, both compounds initially exhibited a prompt propagation phase with a subsequent plateau phase, followed by a rapid degradation phase. The analytical LC/MS method for phosphatidylcholine hydroperoxides might be useful for the analysis of biological samples.


Magnetic resonance safety.
Magnetic resonance imaging (MRI) has a superior soft-tissue contrast compared to other radiological imaging modalities and its physiological and functional applications have led to a significant increase in MRI scans worldwide. A comprehensive MRI safety training to protect patients and other healthcare workers from potential bio-effects and risks of the magnetic fields in an MRI suite is therefore essential. The knowledge of the purpose of safety zones in an MRI suite as well as MRI appropriateness criteria is important for all healthcare professionals who will work in the MRI environment or refer patients for MRI scans. The purpose of this article is to give an overview of current magnetic resonance safety guidelines and discuss the safety risks of magnetic fields in an MRI suite including forces and torque of ferromagnetic objects, tissue heating, peripheral nerve stimulation, and hearing damages. MRI safety and compatibility of implanted devices, MRI scans during pregnancy, and the potential risks of MRI contrast agents will also be discussed, and a comprehensive MRI safety training to avoid fatal accidents in an MRI suite will be presented.


A morphometric study of the lumbar foramen. Influence of flexion-extension movements and of isolated disc collapse.
The authors undertook a morphometric study of the intervertebral foramina in anatomic specimens of spines unaffected by degenerative lesions. They studied the variations in shape and size of 38 intervertebral foramina during flexion-extension movements and following an isolated disc collapse. The cast technique takes into account the bony prominences, the bulge of the disc anteriorly and of the capsulo-ligamentous structures posteriorly. In flexion, all the diameters of the foramina are maximal. In full extension all the diameters decrease significantly: the pedicles come closer together, the disc bulges posteriorly and the ligamentum flavum is pushed forward by the superior articular process of the underlying vertebra. A disc collapse of 4 mm decreases all the diameters, and in this case lumbar extension results in a sufficient decrease of foraminal diameter to threaten the nerve root.


Kerevata Belokobylskij (Hymenoptera: Braconidae: Rogadinae) is no longer a Papua New Guinean endemic with descriptions of three new species from the Indomalayan Region.
The rogadine genus Kerevata Belokobylskij is newly reported from the Indomalayan region. We describe and illustrate three new species, two from India (K. kethai sp. nov. and K. orientalia sp. nov.) and one from Vietnam (K. longi sp. nov.) and provide an illustrated key to the extant species of the genus along with the photographic illustration of the type species of K. pacifica Belokobylskij. Range extension and morphological characters of Kerevata are discussed.


Quantitation of related proteins by Western blot analysis.
A spectrophotometric assay for quantitation of related proteins following Western blot analysis is described. Proteins were separated by polyacrylamide gel electrophoresis and electrophoretically transferred to nitrocellulose. In the system utilized in this study, immobilized interleukin-1 (IL-1) proteins were identified by ELISA reaction using a rabbit antibody specific for IL-1 beta. Subsequently, the membrane was probed with goat anti-rabbit antibody conjugated to horseradish peroxidase and IL-1 proteins were detected by incubation with 4-chloro-1-naphthol and hydrogen peroxide. The stained IL-1 bands were cut out, exposed to the peroxidase chromogenic substrate, o-phenylenediamine (OPD), and hydrogen peroxide and the rate of reaction was determined spectrophotometrically at 490 nM. A linear relationship between enzyme activity and IL-1 concentration was observed from 10 to 1000 ng/lane. Thus, this represents a sensitive and specific method for quantitating small amounts of IL-1 in complex protein mixtures.


Dielectrophoresis of nanocolloids: a molecular dynamics study.
Dielectrophoresis (DEP), the motion of polarizable particles in non-uniform electric fields, has become an important tool for the transport, separation, and characterization of microparticles in biomedical and nanoelectronics research. In this article we present, to our knowledge, the first molecular dynamics simulations of DEP of nanometer-sized colloidal particles. We introduce a simplified model for a polarizable nanoparticle, consisting of a large charged macroion and oppositely charged microions, in an explicit solvent. The model is then used to study DEP motion of the particle at different combinations of temperature and electric field strength. In accord with linear response theory, the particle drift velocities are shown to be proportional to the DEP force. Analysis of the colloid DEP mobility shows a clear time dependence, demonstrating the variation of friction under non-equilibrium. The time dependence of the mobility further results in an apparent weak variation of the DEP displacements with temperature.


Realizing the 2050 Paris climate agreement in West Africa: the role of financial inclusion and green investments.
International organizations have emphasized the importance of global economies supporting efforts to combat climate change. The Paris Agreement or Agenda 2050 urges nations to ensure that the increase in global temperature is limited to 1.5 °C. Studies have analyzed the factors that contribute to harmful emissions, particularly carbon dioxide emissions, in order to limit temperature rise. However, since there are other equally harmful pollutants, this study evaluates the impact of financial inclusion and green investment on reducing greenhouse gas emissions. The study uses data from West Africa, where environmental pollution has significantly increased. The study employed regression analysis while controlling for economic growth, foreign direct investment (FDI), and energy consumption. The study's key findings reveal that financial inclusion and green investment have a monotonic effect on reducing greenhouse gas emissions. Additionally, the study confirms the environmental Kuznets curve hypothesis and the pollution haven effect for the region. Technological innovation reduces pollution, but green investment and financial inclusion reinforce this effect. Therefore, the study recommends that governments in the sub-region commit to supporting green investment and environmentally friendly technological innovations. It is also crucial to strictly enforce laws regulating the operations of multinational corporations in the region.


HRT1 modulates vascular smooth muscle cell proliferation and apoptosis.
The Notch signaling pathway plays vital roles in vascular development and homeostasis. However, the functional role of HRT1, a primary downstream effector of Notch signaling in VSMC, is poorly characterized. In the present study, we postulated that HRT1 plays fundamental roles in modulating VSMC fate. To test the hypothesis that HRT1 is coupled to growth regulation, we generated VSMC lines constitutively overexpressing HRT1 (HRT1SMC) and demonstrated an exaggerated growth behavior compared to its control cell line. The lack of cell cycle arrest at confluence in HRT1SMC was associated with an attenuated up-regulation of the cell cycle inhibitor, p21(WAF1/CIP1). We further established that both transient and constitutive HRT1 signaling promoted VSMC survival in response to serum deprivation and pro-apoptotic Fas ligand. Resistance to apoptosis was associated with the induction of Akt expression/activity, a well-described anti-apoptotic mediator. Overall, these findings provide initial evidence that HRT1 functions as a critical determinant of VSMC proliferation and survival.


Dynamic extreme values modeling and monitoring by means of sea shores water quality biomarkers and valvometry.
Water quality can be evaluated using biomarkers such as tissular enzymatic activities of endemic species. Measurement of molluscs bivalves activity at high frequency (e.g., valvometry) during a long time period is another way to record the animal behavior and to evaluate perturbations of the water quality in real time. As the pollution affects the activity of oysters, we consider the valves opening and closing velocities to monitor the water quality assessment. We propose to model the huge volume of velocity data collected in the framework of valvometry using a new nonparametric extreme values statistical model. The objective is to estimate the tail probabilities and the extreme quantiles of the distribution of valve closing velocity. The tail of the distribution function of valve closing velocity is modeled by a Pareto distribution with parameter 𝜃 t,τ , beyond a threshold τ according to the time t of the experiment. Our modeling approach reveals the dependence between the specific activity of two enzymatic biomarkers (Glutathione-S-transferase and acetylcholinesterase) and the continuous recording of oyster valve velocity, proving the suitability of this tool for water quality assessment. Thus, valvometry allows in real-time in situ analysis of the bivalves behavior and appears as an effective early warning tool in ecological risk assessment and marine environment monitoring.


Correlation between the triglyceride-glucose index and the onset of atrial fibrillation in patients with non-alcoholic fatty liver disease.
Non-alcoholic fatty liver disease (NAFLD) is associated with atrial fibrillation (AF). Insulin resistance (IR) is the main cause of the high prevalence of AF in NAFLD patients. The triglyceride-glucose index (TyG) is a novel IR-related indicator implicated in the incidence and severity of NAFLD. However, the role of TyG in determining the risk for AF in patients with NAFLD remains unclear. A retrospective study was conducted on 912 patients diagnosed with NAFLD via ultrasonography. These patients were divided into two groups: (1) NAFLD+ AF and (2) NAFLD+ non-AF. Least Absolute Shrinkage and Selection Operator (LASSO) regression was used to assess the correlation between the TyG index and the high risk for AF. A receiver operating characteristic (ROC) curve was constructed to evaluate the predictive value for the TyG index for AF. Restricted cubic splines (RCS) were used to test the linear correlation between TyG and the risk for AF. A total of 204 patients with AF and 708 patients without AF were included in this study. The LASSO logistic regression analysis showed that TyG was an independent risk factor for AF (odds ratio [OR] = 4.84, 95% confidence interval [CI] 2.98-7.88, P < 0.001). The RCS showed that the risk for AF increased linearly with TyG over the entire TyG range; this risk was also evident when the patients were analyzed based on sex (P for nonlinear > 0.05). In addition, the correlation between TyG and AF was a consistent finding in subgroup analysis. Furthermore, ROC curve analysis showed that TyG levels combined with traditional risk factors improved the predictive value for atrial fibrillation. The TyG index is useful in assessing the risk for atrial fibrillation in patients with NAFLD. Patients with NAFLD and increased TyG indices have higher risks for atrial fibrillation. Therefore, TyG indices should be assessed when managing patients with NAFLD.


Spanish Pacemaker Registry. 14th Official Report of the Spanish Society of Cardiology Working Group on Cardiac Pacing (2016).
This report describes the results of analysis of implanted pacemakers reported to the Spanish Pacemaker Registry. The analysis was based on information provided by the European Pacemaker Identification Card. Information was received from 115 hospitals, with a total of 12 697 cards, representing 32.3% of the estimated activity. Use of conventional and resynchronization pacemakers was 818 and 79 units per million inhabitants, respectively. A total of 200 leadless pacemakers were implanted. The mean age of the patients receiving an implant was 77.8 years, and 52% of devices were implanted in persons older than 80 years. In all, 74.9% were first implants and 23.4% corresponded to generator exchange. Endocardial leads were bipolar, 82.9% with active fixation, and 16.1% had magnetic resonance imaging protection. Most patients received bicameral sequential pacing, although single chamber pacing VVI(R) was used in 26.7% of the patients with sick sinus syndrome and in 23.8% of those with atrioventricular block, despite sinus rhythm. Total use of pacemaker generators in Spain has increased by 1.6% compared with 2015. Most implanted leads have active fixation and less than 20% have magnetic resonance imaging protection. Age and sex directly influenced pacing mode selection, which could be improved in around 32% of patients.


Cyclic fertilin-derived peptide stimulates in vitro human embryo development.
To study the cyclic fertilin peptide effects on preimplantation human embryogenesis. Cyclic fertilin peptide reproduces the structure of the binding site of the sperm Fertilin β (also named A Disintegrin and Metalloprotease 2: ADAM2) disintegrin domain. It binds to the oocyte membrane and increases sperm-oocyte fusion index in human and fertilization rate in mouse, providing healthy pups. It also improves human oocyte maturation and chromosome segregation in meiosis I and binds to human embryo blastomeres, suggesting that it has a membrane receptor. Thawed human embryos at the 3 to 4 cells stage were randomly included in a dose-response study with cyclic fertilin peptide. Inner cell mass (ICM), trophectoderm (TE), and total cell numbers were evaluated in top- and good-quality blastocysts. The study was performed in an academic hospital and research laboratory. Human embryos donated for research. This project was approved by the French "Agence de la Biomédecine." Immunofluorescence and tissue-specific gene expression analysis, using Clariom D microarrays, were performed to study its mechanism of action. Cyclic fertilin peptide improves blastocyst formation by almost 20%, the concentration of 1 μM being the lowest most efficient concentration. It significantly increases twice the TE cell number, without modifying the ICM. It increases the in vitro hatching rate from 14% to 45%. Cyclic fertilin peptide stimulates TE growth. In the ICM, it induces transcriptional activation of intracellular protein and vesicle-mediated transport. Cyclic fertilin peptide dramatically improves human embryo development potential. It could be used to supplement culture medium and improve the in vitro human embryo development. Starting supplementation immediately after fertilization, instead of day 2, could significantly upgrade assisted reproductive technology outcome.


Millimeter wave reflectivity used for measurement of skin hydration with different moisturizers.
A new non-invasive method for determining the free water content in human skin has been developed. The method analyzes the reflection of millimeter (mm) wavelength electromagnetic waves. The amount of reflection of mm waves depends on an electrical property (namely, the permittivity) of the skin, and this depends upon the free water content of the various skin layers. The aim of the present study was to use the mm wave reflectometry method for determination of free water content in healthy skin treated with different hydrating substances. Skin lotion, pure water, glycerol, and petroleum jelly (an occlusive substance) were used for hydration of skin. The amount of free water was calculated using the permittivity values of skin layers found from fitting a three layer skin model to measured reflection data. The skin model consisted of (1) the stratum corneum (SC), (2) the viable epidermis plus the dermis, and (3) fat layers. Mm wave reflection was significantly affected by the water content of the thick SC of the palm but not by the very thin SC of the forearm. Treatment of the forearm and palm skin with different hydrating substances produced notable changes of the free water content in the SC, but not in the viable epidermis or dermis. The greatest hydration was produced by pure water and skin lotion, and the lowest by petroleum jelly. However, petroleum jelly produced prolonged retention of water in the SC following its hydration by other moisturizers. The content of free water was found to return to its baseline value after removal of moisturizers in as short a time as 8.3 min. The study shows that mm wave reflectometry can be used as a sensitive technique for the non-invasive determination of water content in living skin.


Sensitizing effect of tacrine on M-cholinergic receptors in gastric smooth muscle of rats.
To find if tacrine exerts a sensitizing effect on the cholinergic receptors of gastric smooth muscles, and study some of the mechanisms inducing it and measure the relative intensity of tacrine's effects on contractile activity. Isometric recording of the mechanical activity of gastric smooth muscle preparations; determination of acetyl-cholinesterase activity in smooth-muscle tissue homogenates using Ellman's method. We found that the threshold concentration for tacrine not reducing the acetylcholinesterase activity and not having an effect on the smooth muscle preparations was 1 x 10(-8) mol/l. This concentration, however, significantly increased the acetylcholine-induced contraction compared with the controls, after the smooth-muscle tissue was incubated for 60 or 100 min. Treating smooth-muscle preparations with tacrine in a concentration of 5 x 10(-6) mol/l triggered a contraction induced by the drug's anti-cholinesterase activity. A secondary contraction was induced after 38.6 +/- 5.6 min. There was no secondary contraction after the control acetylcholine-induced effect. Atropine (1 x 10(-6) mol/l) inhibits this effect. Preliminary treatment of smooth muscle preparations with hexamethonium (1 x 10(-6) mol/l) did not change significantly the intensity of the first phase of tacrine-induced contraction and shifted in time the appearance of the second contractile phase. Tacrine has a sensitizing effect on M-cholinergic receptors; it occurs after a long incubation of the gastric smooth muscles with the drug and is manifested as a secondary contraction which is shifted in time and is significantly inhibited by atropine.


Synergistic Combination of Facile Thiol-Maleimide Derivatization and Supramolecular Solvent-Based Microextraction for UHPLC-HRMS Analysis of Glutathione in Biofluids.
Glutathione (GSH) is the most abundant non-protein thiol in biofluids, enabling diverse physiological functions. Among the proposed methods for GSH detection, ultra-high-performance liquid chromatography (UHPLC) coupled with high-resolution mass spectrometry (HRMS) has the advantages of high sensitivity and efficiency. In this study, a novel analytical method was developed for the determination of GSH using supramolecular solvent (SUPRAS)-based dispersive liquid-liquid microextraction (DLLME) and UHPLC-HRMS. N-Laurylmaleimide was dissolved in tetrahydrofuran, which served three functions: 1) precipitate the proteins present in the biofluid sample, 2) provide a reaction environment for derivatization, and 3) enable the use of SUPRAS as the dispersing agent. Critical parameters were optimized based on single factor testing and response surface methodology. The established method was validated in terms of linearity, accuracy, precision, and successful quantitative analysis of GSH in saliva, urine, and plasma samples. Experimental results showed that SUPRAS as an extraction solvent was particularly suitable for the extraction of GSH from complex matrices. The current study provides a useful tool for accurate measurements of GSH concentrations, which could potentially be used for clinical diagnostics.


Chemosensitization at reduced nitroimidazole concentrations by mixed-function compounds combining 2-nitroimidazole and chloroethylnitrosourea.
A mixed-function compound (I-278) combining 2-nitroimidazole and chloroethylnitrosourea has been shown to be greater than 2-fold more toxic to hypoxic HeLa-MR cells than to cells similarly exposed under aerobic conditions, consistent with chemosensitization of nitrosourea toxicity by the 2-nitroimidazole Misonidazole (MISO). However, in the case of I-278, the enhancement resulted from micromolar concentrations of 2-nitroimidazole as opposed to the millimolar quantities required for a similar enhancement by MISO. These experiments provide evidence (1) that the enhanced hypoxic toxicity of I-278 is not attributable to additional, independent hypoxic cell killing by the nitroimidazole group and (2) that the interaction between the two functions under hypoxic conditions results in increased crosslink formation typical of chemosensitization. The data strongly suggest that the chemosensitizing efficiency of nitroimidazoles can be dramatically improved by covalent linkage to a chloroethylating species.


Transparent intrastromal corneal lenticule obtained from SMILE surgery as a free graft for the treatment of primary pterygium: A pilot study.
To report our initial experience in patients with primary pterygium surgery who had a transparent intrastromal corneal lenticule as a free graft obtained during small incision lenticule extraction (SMILE) surgery. This study enrolled five eyes of 5 patients with pterygium who were surgically treated with single or double free lenticule that was sutured to the defective area of the nasal corneosclera immediately following pterygium excision. All cases with pterygium were primary, and topical mitomycin C was not used at the time of surgery. The mean age of the patients was 51.0 years and 2 of the 5 eyes belonged to male subjects. The patients were followed up for an average of six months. During the follow-up period, the lenticule grafts demonstrated no sign of rejection and were intact in all cases. All patients recovered well with no complications or recurrences at six months postoperatively. This report suggests that transparent corneal lenticules obtained during SMILE laser surgery may be used as an alternative novel graft source for the surgical treatment of patients with primary pterygium. It appears to be a safe, easy, cost- and time-effective reliable method.


Mediation of serotonin hyperalgesia by the cAMP second messenger system.
In this study we have evaluated the second messenger system that might couple 5-HT1A receptor activation to produce peripheral hyperalgesia. The intradermal injection of the serotonin (5-hydroxytryptamine; 5-HT) receptor agonist for the 1A receptor subset (5-HT1A), (+/-)-2-dipropylamino-8-hydroxy-1,2,3,4-tetrahydronaphthaline hydrobromide (8-OH DPAT) produces a dose-dependent hyperalgesia which was attenuated by a cAMP kinase inhibitor (the R-isomer of cyclic adenosine-3'-5'-monophosphate), but prolonged by the inhibition of endogenous phosphodiesterase by rolipram, supporting a role for the cAMP second messenger system. The 5-HT1A receptor agonist, 8-OH-DPAT, and the adenyl cyclase activator, forskolin administered together, produced an additive hyperalgesia, suggesting that the 5-HT1A receptor in peripheral terminals of the primary afferent neurons is positively coupled to the cAMP second messenger system in producing hyperalgesia. The inability of pertussis toxin to inhibit 8-OH DPAT-induced hyperalgesia further supports this hypothesis. The coupling of the 5-HT1A receptor to the cAMP second messenger system appears to be through guanine regulatory proteins since guanosine 5'-O-(3-thiotriphosphate) and cholera toxin both markedly enhanced 8-OH DPAT hyperalgesia. In further support of the role of guanine nucleotide regulatory proteins, guanosine 5'-O-(2-thiodiphosphate), as well as activators of inhibitory guanine regulatory proteins (the mu-opioid agonist, [D-Ala2,N-Me-Phe4,Gly5-ol]-enkephalin, and the adenosine A1 agonist, N6-cyclopentyladenosine, significantly attenuated 8-OH DPAT hyperalgesia.


[Maxillofacial dysmorphosis and myasthenia. Orthodontic and surgical treatment of a case].
Case report of a patient with myasthenia and dysmorphic facial features. Pluridisciplinary treatments approach.


Adhesion molecules involved in protein kinase A- and C-dependent lymphocyte adherence to microvascular endothelial cells.
A twofold increase in lymphocyte adherence to rat microvascular endothelial cells (EC) was achieved by incubating EC for 4 h with IL-1 alpha or dibutyryl-cAMP (stimulators of protein kinase A, PKA) and PMA (stimulator of protein kinase C, PKC). Monoclonal antibodies anti-CD11a, anti-CD18 (LFA-1) and anti-CD49d (VLA-4 alpha) significantly inhibited the increased lymphocyte binding to IL-1 alpha-induced EC, anti-CD18 and to a lesser extent anti-CD11a and anti-CD49d to dibutyryl-cAMP-induced EC, whereas only anti-CD11a and anti-CD18 monoclonal antibodies inhibited PMA-induced lymphocyte binding. These findings suggest that stimulation of PKA and PKC induces lymphocyte binding to EC via different adhesion molecules.


Trends in market demand for internal medicine 1999 to 2004: an analysis of physician job advertisements.
The health care marketplace has changed substantially since the last assessment of demand for internal medicine physicians in 1996. We reviewed internal medicine employment advertisements published in 4 major medical journals between 1996 and 2004. The number of positions, specialty, and other practice characteristics (e.g., location) were collected from each advertisement. Four thousand two hundred twenty-four advertisements posted 4,992 positions. Of these positions, jobs in the Northeast (31% of positions) or single specialty groups (36.8% of positions) were most common. The relative proportion of advertisements for nephrologists declined (P < .001), while the relative proportions of advertisements for critical care specialists (0.5% in 1996 to 1.7% in 2004, P = .004) and hospitalists (1.0% in 1996 to 12.1% in 2004, P < .001) increased. Advertisements for outpatient-based generalist positions (i.e., Primary Care and Internal Medicine) declined (-2.7% relative annual change, 95% confidence interval [95% CI] -4.1%, -1.2%) between 1996 and 2004, a decrease largely due to a substantial decline in advertisements noted between 1996 and 1998. However, over the entire time period, the combined proportion of advertisements for all generalists (hospitalists and outpatient-based generalists) did not change (0.5% relative annual change, 95% CI -0.8% to 2.0%). Since 1996, demand for the majority of medical subspecialties has remained constant while relative demand has decreased for primary care and increased for hospitalists and critical care. Increase in demand for generalist-trained hospitalists appears to have offset falling demand for outpatient generalists.


Immunocytochemical assays in human endometrial carcinomas: a multiparametric computerized analysis and comparison with nonmalignant changes.
Immunocytochemical assay (ICAs) were performed on frozen sections from human endometrial samples (n = 89) including normal endometrium, decidua, hyperplasia with and without atypia, and carcinomas. Monoclonal antiestrogen receptor (ER), anti-laminin (Lam), anti-type IV collagen (Coll IV), and anti-Ki67 were applied with avidin-biotin-peroxidase complex or peroxidase-antiperoxidase complex. The results of the ICAs were evaluated through a computerized system of image analysis referred to as SAMBA. It was shown that this system provided for an accurate reliable and reproducible analysis of ICAs in tissue sections. It is concluded that this multiparametric and standardized method of analysis of ICAs can further be applied in correlations with clinical and biochemical data.


Association of metabolic health phenotypes, obesity, and hepatocellular carcinoma risk.
The obesity and hepatocellular carcinoma (HCC) risk association may differ by individuals' metabolic health status. To investigate the association between obesity categories and HCC risk among individuals with different metabolic health phenotypes. A case-control study among 518 HCC cases and 1,036 frequency-matched controls was conducted. Body mass index (BMI) was assessed before diagnosis. Pre-diagnosis data on dyslipidemia, hypertension, and diabetes were used to categorize participants as metabolically healthy or metabolically unhealthy. Participants were further categorized into metabolically healthy normal weight (MHNW), metabolically healthy overweight (MHOW), metabolically healthy obese (MHO), metabolically unhealthy normal weight (MUNW), metabolically unhealthy overweight (MUOW), and metabolically unhealthy obese (MHO). We used logistic regression to calculate multivariable-adjusted odds ratios (ORs) and 95% confidence intervals (CIs). Being overweight (OR=1.68, 95%CI=1.21-2.34) or obese (OR=1.49, 95%CI=1.11-1.89) was associated with higher HCC risk. Among metabolically healthy participants, no association was found between being overweight or obese and HCC risk. However, among the metabolically unhealthy participants, being overweight (OR=1.89, 95%CI=1.31-2.72) or obese (OR=1.50, 95%CI=1.07-2.09) was associated with higher HCC risk. Compared to the MHNW phenotype, no association was found between the MHOW and MHO phenotypes and HCC risk, but the MUNW (OR=1.94, 95%CI=1.09-3.43), MUOW (OR=3.78, 95%CI=2.15-6.65), and MUO (OR=2.93, 95%CI=1.70-5.05) phenotypes were associated with higher HCC risk. The association between BMI and HCC appears to be restricted to individuals with underlying metabolic abnormalities.


The effects of neurochemical lesioning of dopaminergic terminals in early ontogenesis on behavior in adult rats.
6-Hydroxydopamine, which induces selective degeneration of the dopaminergic system of the brain, was given intraamniotically to rats on days 13 and 17 of intrauterine development at a dose of 75 microg/fetus. Similar experiments were performed with 6-hydroxydopamine on days 4 and 10 of neonatal life. Rats were subsequently reared and motor and emotional (dopamine-dependent) types of behavior were studied in adulthood, addressing behavior in the open field test, rotatory behavior, anxiety in an elevated cross maze, a place-preference conditioned response, acquisition of the ability to differentiate new and old arms in a Y maze, aggressivity in the "foreigner-resident" test, and self-stimulation in a Skinner box. Prenatal exposure, to a lesser extent than postnatal exposure, initiated rotatory and stereotypical behavior, decreased the level of anxiety (fear) in the elevated maze, and reinforced the effects of phenamine in the conditioned place-preference test, impaired the differentiation of old and new Y-maze arms, impaired aggressivity in the "foreigner-resident" test, and impaired self-stimulation of the lateral hypothalamus. It is concluded that the early postnatal period of development is more sensitive to the action of this neurotoxin than the prenatal period. This is associated with the critical periods of the formation of the dopaminergic system during ontogenesis, which depend on synaptogenesis.


Brucella abortus induces apoptosis of human T lymphocytes.
Immune evasion is essential for Brucella abortus to survive in the face of robust adaptive CD4+ T cell response. We have previously demonstrated that B. abortus can indirectly inhibit CD4+ T cells by down-regulating MHC-II expression and antigen presentation on macrophages. However, whether B. abortus is able to directly interfere with T lymphocytes is not known. We report here that B. abortus induces apoptosis of human T lymphocytes, even though invasion of T lymphocytes was low and non-replicative. The ability of heat-killed B. abortus to reproduce the same phenomenon suggested that there was a bacterial structural component involved. We demonstrated that a prototypical B. abortus outer membrane lipoprotein (l-Omp19), but not its unlipidated form, induced T lymphocyte apoptosis. Moreover, a synthetic lipohexapeptide that mimics the structure of the protein lipid moiety also induced an increase in T lymphocyte cell death, indicating that the structural component implicated in the phenomenon could be any B. abortus lipoprotein. B. abortus-induced T lymphocyte apoptosis was dependent on the secretion of TNF-α since pre-incubation of T lymphocytes with anti-TNF-α mAb inhibited the apoptosis of the cells. Overall, these results represent a new mechanism whereby B. abortus by directly inhibiting T cell-mediated responses may evade adaptive immune responses.


Prognostic Value of HER-2/neu Gene Amplification in Epithelial Ovarian Carcinoma.
Human epidermal growth factor receptor 2 (HER2) has tyrosine kinase activity and is a member of the epidermal growth factor receptor family. This initiates a variety of signal for pathways which leads to cell proliferation and tumourigenesis. In approximately 15-30% of breast cancers and 10-30% of gastric/gastroesophageal cancers, amplification or overexpression of HER2 occurs, and it serves as a prognostic and predictive biomarker. HER-2/neu is one of the most frequently studied molecular biological parameters in epithelial ovarian cancer (EOC), but its prognostic impact has not been fully assessed. The objective of the current study was firstly to analyse the presence of HER-2 overexpression in EOC patients and secondly to evaluate association between human epidermal growth factor receptor 2 (HER-2/neu) expression and progression-free survival in patients with EOC. In this prospective study of 2 years in our department, 186 gynaecological cancers were operated out of which 98 cases were operated for epithelial ovarian cancer and rest for cervical cancer, endometrial cancer, and other subtypes of ovarian cancer. In this study, HER2 gene status was evaluated in EOC by immunohistochemistry analysis, and overall survival of these patients was evaluated. HER-2 overexpression was found in 22 of the 98 investigated cases (22.45%), in which 14 OC (14.29%) were weakly positive and 8 OC (8.16%) were moderately to intensely positive. In the study, increased HER2 expression was associated with decreased progression-free survival (PFS) and hence poor prognosis (P 0.054). The introduction of HER2-directed therapies has dramatically influenced the outcome of patients with HER2 positive breast and gastric/gastroesophageal cancers. The present study findings provided that HER-2/neu expression in patients with OC has an adverse impact on the PFS. Therefore, our results show that the decision algorithm usually used in breast cancer by HER2 may be appropriate in ovarian cancer.


Do 25-Hydroxyvitamin D Levels Correlate With Fracture Complications?
To determine the relationship between serum 25-hydroxyvitamin D [25(OH)D] levels and the likelihood of postoperative complications and fracture reoperation rate in orthopaedic trauma patients receiving vitamin D and calcium supplementation. Retrospective case series. Level I trauma center, Midwestern United States. All orthopaedic trauma patients-18 years or older-over a 20-month period were included with available initial and repeat 25(OH)D serum levels. In total, 201 patients met inclusion criteria. All patients received 1000 IU of vitamin D3 and 1500 mg of calcium daily. Vitamin D deficient and insufficient patients also received 50,000 IU of ergocalciferol (vitamin D2) weekly until 25(OH)D levels normalized or fractures healed. fracture complications and 25(OH)D levels. Fifteen patients experienced postoperative healing complications. There was no significant difference between initial (P = 0.92) or repeat (P = 0.91) 25(OH)D levels between patients with and without fracture healing complications. Twenty-eight patients required repeat orthopaedic surgery. There was no significant difference between initial (P = 0.62) or repeat (P = 0.18) 25(OH)D levels between patients who did or did not require repeat orthopaedic surgery. There was no significant difference between initial (P = 0.66) or repeat (P = 0.89) 25(OH)D levels between patients who did or did not require nonorthopaedic surgery. Serum 25(OH)D levels did not significantly affect the likelihood of fracture healing complications requiring surgery or any nonorthopaedic injury-related surgery. Therapeutic Level IV. See Instructions for Authors for a complete description of levels of evidence.


Expression, purification and preliminary biochemical and structural characterization of the leucine rich repeat namesake domain of leucine rich repeat kinase 2.
Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial Parkinson's disease. Much research effort has been directed towards the catalytic core region of LRRK2 composed of GTPase (ROC, Ras of complex proteins) and kinase domains and a connecting COR (C-terminus of ROC) domain. In contrast, the precise functions of the protein-protein interaction domains, such as the leucine-rich repeat (LRR) domain, are not known. In the present study, we modeled the LRRK2 LRR domain (LRR(LRRK2)) using a template assembly approach, revealing the presence of 14 LRRs. Next, we focused on the expression and purification of LRR(LRRK2) in Escherichia coli. Buffer optimization revealed that the protein requires the presence of a zwitterionic detergent, namely Empigen BB, during solubilization and the subsequent purification and characterization steps. This indicates that the detergent captures the hydrophobic surface patches of LRR(LRRK2) thereby suppressing its aggregation. Circular dichroism (CD) spectroscopy measured 18% α-helices and 21% β-sheets, consistent with predictions from the homology model. Size exclusion chromatography (SEC) and dynamic light scattering measurements showed the presence of a single species, with a Stokes radius corresponding to the model dimensions of a protein monomer. Furthermore, no obvious LRR(LRRK2) multimerization was detected via cross-linking studies. Finally, the LRR(LRRK2) clinical mutations did not influence LRR(LRRK2) secondary, tertiary or quaternary structure as determined via SEC and CD spectroscopy. We therefore conclude that these mutations are likely to affect putative LRR(LRRK2) inter- and intramolecular interactions.


Health Profession Students' Awareness, Knowledge, and Confidence Regarding Preexposure Prophylaxis: Results of a National, Multidisciplinary Survey.
Preexposure prophylaxis (PrEP) is a highly effective, pharmacologic method of HIV prevention. Despite its safety and efficacy, PrEP prescription remains low in those patients who are at highest risk for HIV infection. One possible reason for this may be the lack of inclusion of PrEP and HIV prevention discussions within the curricula of health professions education. An online survey was administered to a cross-sectional sample of future prescribers (osteopathic/allopathic medical and physician assistant students), future nurses, and future pharmacists (n = 2085) in the United States between January and July 2019 to assess and compare awareness of PrEP, PrEP education, PrEP knowledge, and confidence in 2 areas related to PrEP. We show that, overall, awareness of PrEP is high among future health care providers (81.6%), with the future pharmacists reporting the greatest awareness (92.2%; P < 0.001) and more commonly reporting PrEP education (71.0%). Students had mixed knowledge of PrEP, with future pharmacists reporting the highest knowledge of PrEP. Approximately 30% of students in all disciplines reported having low confidence counseling a patient about PrEP and low confidence educating a colleague about PrEP. Knowledge of PrEP was a significant predictor of confidence counseling a patient about PrEP (P < 0.001) and educating a colleague about PrEP (P < 0.001). This study identifies opportunities to improve and incorporate evidence-based strategies for educating future health care providers about PrEP for HIV prevention within health professions curricula.