Iran's higher education and COVID-19: the "new normal" for Tehran University of Medical Sciences.
When the emergency committee of the World Health Organization declared that the outbreak of COVID-19 meets the criteria of a "Public Health Emergency of International Concern" on January 30th, 2020, no one could ever imagine how soon it will spread globally, and a health crisis would turn to a social crisis that affects everything including higher education. However, during this uncertainty, Tehran University of Medical Sciences (TUMS) tried to respond quickly. In this study, we explain how a nerve center has helped TUMS respond to this crisis and ensure safety to keep key operations going, and set up a functional decision-making system for the future. We also share perspectives on the critical issues, the challenges ahead, and the opportunities emerging in the "new normal."


Ceramic inlay systems: some clinical aspects.
Due to their inertness and unsurpassed aesthetic properties, ceramic materials are increasingly used as inlay materials. Based on a questionnaire, the frequency and character of complications related to ceramic inlay therapy were assessed and some important pitfalls and clinical relevant factors discussed. A majority of the dentists performed between 1 and 10 ceramic inlays per month using several different ceramic materials. As many as 85% of the dentists had observed complications in connection with ceramic inlays. Most frequency reported was hypersensitivity of a transient nature (15%). The material revealed an overall inlay fracture frequency of 5%, but the individual variations were high. Tooth fracture, loss of retention and other complications such as secondary caries, endodontic envolvement, colour mismatch and a surface and edge deterioration occurred with a varying but lower frequency. Failure analysis revealed a large number of causative factors among which cavity and inlay design, bonding procedures, selection of luting material and proper case selection were regarded as the most important. Due consideration to the strength of remaining tooth substance was found to be important in reducing the problem of hypersensitivity and the risk of tooth and inlay fracture.


Acid exposure enhances sporulation of certain strains of Clostridium perfringens.
A gastroenteritis results when Clostridium perfringens is ingested in high numbers and sporulates releasing enterotoxin in the intestines. Since the organism must pass through the stomach, its ability to form spores may be affected by the acidic environment. Five strains of C. perfringens were exposed to acidic conditions and then assessed for survival and their ability to form spores. An acidic pH environment kills the bacteria over time but surviving cells are able to recover and form spores. Two of the five strains demonstrated enhanced sporulation following a 30-min exposure to a pH 2 environment. For four of the strains tested, enterotoxin concentrations were higher from acid-exposed cells than from untreated cells. Exposure to a pH 3.5 environment did not affect sporulation when compared to an untreated control. Bacteria in the stationary phase of growth were the most able to resist the acid and sporulate. The results indicate that some strains will produce more spores and enterotoxin following exposure to an acidic environment.


Expression of N-cadherin by skeletal muscle in the degeneration and the degeneration/regeneration processes after nerve injury.
We investigated the expression of N-cadherin by skeletal muscle during the degeneration and degeneration/regeneration processes using the rat sciatic nerve and gastrocnemius muscle model. The right sciatic nerve was exposed in the mid-thigh region, and the nerve was transsected with small scissors. After then, nerve was sutured (sutured group), or both edges of the resected nerve were turned and sutured to the muscle of each side (unsutured group). At various periods up to 24 weeks after the operation the middle portion of the gastrocnemius muscle of the treated hindlimbs was removed. Expression of N-cadherin was detected by western blot analysis and immunofluorescent staining with an anti N-cadherin antibody. In the both groups, the degree of expression had already increased by the end of the first postoperative week, but there were no significant differences between the first and second postoperative weeks between the two groups. However, the values recorded at the fourth, sixth, ninth, and twelfth postoperative weeks were significantly higher in the unsutured group than in the sutured group. Immunofluorescent staining was present around the muscular membrane in all specimens including the control. These results indicated that there was a difference in the kinetics of expression of N-cadherin in skeletal muscle between the degeneration and degeneration/regeneration processes of the muscle after injury to the nerve. It was also clear that N-cadherin has a role at the surface of the muscle cell in skeletal muscle, not in the satellite and inflammatory cells, in both groups.


Cell migration and metastasis markers as targets of environmental pollutants and the Aryl hydrocarbon receptor.
During the last few years, several studies have pointed to a surprising link between environmental pollutants cellular signaling and important cell functions such as plasticity, adhesion and migration. This unexpected link could be related to endogenous functions of pollutants receptors that may be disrupted by environmental factors, which is supported by observations in invertebrate species. It could also reveal novel toxic end-points and mechanisms of those pollutants, such as teratogenesis and cancer metastasis that are highly relevant from a public health point of view. In the present short article, we will review our recent observations on the aryl hydrocarbon receptor and its new molecular and cellular targets. We identified HEF1/NEDD9/CAS-L, a multifunctional protein involved in integrin-based signaling as a transcriptional target of the receptor, and showed that its induction was critical for cell plasticity mediated by environmental pollutants. We will put our studies in perspective with other observations made by several groups.


Serum symmetric dimethylarginine and creatinine in Birman cats compared with cats of other breeds.
Objectives The aim of this study was to assess whether, in contrast to serum creatinine, which is higher in Birman cats than in other breeds, the serum concentration of symmetric dimethylarginine (SDMA) is comparable in clinically healthy Birmans and in the general feline population. This could allow, in this breed, to better evaluate chronic kidney disease (CKD). Methods Serum creatinine and SDMA were measured in clinically healthy Birmans (n = 50) and in cats of other breeds (n = 46), and the results were statistically compared. A breed-specific reference interval (RI) was established for Birmans and compared with the RI for the general feline population (0.0-14.0 µg/dl). Results Creatinine (1.58 ± 0.36 mg/dl) and SDMA (12.2 ± 2.8 µg/dl) were higher ( P <0.001) in Birmans than in cats of other breeds (1.19 ± 0.17 mg/dl; 10.3 ± 2.5 µg/dl). In 20/50 Birman cats (40.0%) serum creatinine was higher than both the non-breed-specific RI of our laboratory and the threshold recommended to classify cats as IRIS stage 2 (1.6 mg/dl). The concentration of SDMA was higher than the pre-existing RI in 10/50 Birmans (20.0%) and in four cats of other breeds (8.7%). Among Birmans, the proportion of cats with SDMA >14 µg/dl was lower ( P <0.017) than the proportion of cats with creatinine >1.60 mg/dl. However, the deviation from the upper limit of the RI was lower than the analytical variability of the method in 7/10 Birmans and in 4/4 cats of other breeds. The breed-specific RI (3.5-18.7 µg/dl) overlapped with the pre-existing one. Conclusions and relevance SDMA may be a better marker of CKD in Birman cats than creatinine when non-breed-specific RIs are utilised. The coupled analysis of creatinine and SDMA could help prevent errors in diagnosing and staging CKD in Birman cats.


The economic impact of failures in total hip replacement surgery: 28,997 cases from the Norwegian Arthroplasty Register, 1987-1993.
The Norwegian Arthroplasty Register was established in 1987. Until January 1994, approximately 200 different implant combinations had been used in total hip replacements (THR) in Norway. About 5,500 THR were performed each year in this period with a total cost of 70 million USD per year. We analyzed the economic consequences related to the use of some inferior primary hip arthroplasties in this period. As the reference arthroplasty, we chose the most commonly used prosthesis in Norway, i.e., the Charnley prosthesis fixed with high viscosity cement containing antibiotic and with systemic antibiotic prophylaxis (n 4,970). We compared this reference group to all other primary THR registered in the same time period (n 24,027), and to the following sub-groups of primary THR: 1) uncemented Ti-Fit/ Bio-Fit (acetabulum/femur) combination (n 173), 2) uncemented Coxa/Femora combination (n 153), 3) THR with low-viscosity cement (n 1,807) and 4) THR with Boneloc cement (n 1,250). We estimated the number of additional revisions compared to the reference arthroplasty after a follow-up of 3-5 years in the different groups, with adjustment for age, sex and diagnosis. The direct extra revision costs were calculated. Compared to the reference arthroplasty, the group of all other primary THR gave an extra revision cost estimated at about 1.7 million USD per year. About 1,000 uncemented Bio-Fit femoral prostheses have been applied in Norway, including those implanted before the registration started (1985-1987). The extra revision costs the first postoperative years for these 1,000 prostheses amount to about 0.7 million USD per year. Corresponding figures in the Coxa/ Femora group were 0.08 million USD, in the group with low-viscosity cement, 0.3 million USD and in the Boneloc group, 0.4 million USD per year.


Concussion Care Practices and Utilization of Evidence-Based Guidelines in the Evaluation and Management of Concussion: A Survey of New England Emergency Departments.
Evidence-based clinical practice guidelines can facilitate proper evaluation and management of concussions in the emergency department (ED), often the initial and primary point of contact for concussion care. There is no universally adopted set of guidelines for concussion management, and extant evidence suggests that there may be variability in concussion care practices and limited application of clinical practice guidelines in the ED. This study surveyed EDs throughout New England to examine current practices of concussion care and utilization of evidence-based clinical practice guidelines in the evaluation and management of concussions. In 2013, a 32-item online survey was e-mailed to 149/168 EDs throughout New England (Connecticut, Rhode Island, Massachusetts, Vermont, New Hampshire, Maine). Respondents included senior administrators asked to report on their EDs use of clinical practice guidelines, neuroimaging decision-making, and discharge instructions for concussion management. Of the 72/78 respondents included, 35% reported absence of clinical practice guidelines, and 57% reported inconsistency in the type of guidelines used. Practitioner preference guided neuroimaging decision-making for 57%. Although 94% provided written discharge instructions, there was inconsistency in the recommended time frame for follow-up care (13% provided no specific time frame), the referral specialist to be seen (25% did not recommend any specialist), and return to activity instructions were inconsistent. There is much variability in concussion care practices and application of evidence-based clinical practice guidelines in the evaluation and management of concussions in New England EDs. Knowledge translational efforts will be critical to improve concussion management in the ED setting.


The clinical significance of serum and urinary neopterin levels in several renal diseases.
Neopterin is a pyrazino-pyrimidine compound, and is known to be a marker associated with cell-mediated immunity in various diseases. We hypothesized that the levels of serum and urine neopterin would be elevated in renal disease, and would correlate with certain clinical parameters. We evaluated serum and urinary neopterin levels in patients with several renal diseases, including nephrotic syndrome (NS, n=19), chronic renal failure (CRF, n=8), end stage renal disease (ESRD, n=64) and controls (n=22). Serum neopterin was elevated in patients with CRF and ESRD, as compared to controls. Urinary neopterin levels were also found to be elevated in patients with CRF and ESRD, as compared to controls. Serum neopterin levels showed significant positive correlation with age, serum BUN and creatinine levels, and inverse correlation with WBC, hemoglobin, hematocrit, serum albumin and total iron binding capacity. Urine neopterin levels exhibited positive correlation with age and serum creatinine levels, and inverse correlation with WBC, hemoglobin, hematocrit, BUN and serum albumin. In conclusion, increased serum and urinary neopterin levels were found in some patients with renal disease and were correlated with certain clinical parameters.


Low values on 50 gram glucose challenge test or oral 100 gram glucose tolerance test are associated with good perinatal outcome.
We set out to reevaluate the hypothesis that high normal (negative) results of 50 g oral glucose challenge test or high normal glucose level on 100 g oral glucose tolerance test are associated with complications of pregnancy and delivery. This was a prospective study involving 735 nondiabetic women. The first group (n=352) was made up of pregnant women with normal 50 g oral glucose challenge test without previous history of diabetes mellitus or gestational diabetes. The second group (n=383) was made up of pregnant women without previous history of diabetes mellitus or gestational diabetes with an abnormal 50 g oral glucose challenge test and with normal 100 g oral glucose tolerance test and not more than one previous delivery. In nondiabetic women, we demonstrated a positive correlation between high normal 50 g glucose challenge test values and the incidence of preeclampsia, caesarean section rate, macrosomia, neonatal hyperlipidaemia and minor congenital abnormalities. We failed to confirm any relationship to any pregnancy complication in pregnant women with 2-hour glucose levels in the range 6.7-9.1 mmol/l on the 100 g oral glucose tolerance test. We have demonstrated a positive relationship between the incidence of premature rupture of membranes and 1-hour glucose level, caesarean section rate and maternal 1-hour glucose level or 1-hour glucose level minus fasting glucose level of 4.2 mmol/l, instrumental delivery rate and maternal 3-hour glucose level, incidence of neonatal macrosomia and 1-hour glucose level, and incidence of neonatal hyperlipidaemia and at least one high but normal glucose level on the 100 g oral glucose tolerance test. With regard to pregnancy and delivery complications there were no significant difference if the high normal value is on the 50 g glucose challenge test or on the 100 g oral glucose tolerance test. It is concluded that one high normal 100 g oral glucose tolerance test or high normal 50 g glucose challenge test are associated with adverse pregnancy and delivery outcome. Nondiabetic women with 50 g glucose challenge test value of 6.1 mmol/l and/or 100 g oral glucose tolerance test values of 5 mmol/l have a favourable pregnancy and delivery outcome.


Modification of the Highly Conductive PEDOT:PSS Layer for Use in Silver Nanogrid Electrodes for Flexible Inverted Polymer Solar Cells.
Silver nanogrid based flexible transparent electrode is recognized as the most promising alternative to ITO electrode for organic electronics, owing to its low production cost and excellent flexibility. Typically, a highly conductive thin film coating layer, such as highly conductive PEDOT:PSS (HC-PEDOT:PSS) is usually deposited onto the Ag-grid electrode to smooth the surface and to minimize the sheet resistance. In this paper, we found that inverted flexible polymer solar cells with structure of Ag-grid/HC-PEDOT:PSS/ZnO/photoactive layer/MoO3/Al generally exhibits strong S-shaped J-V curves, which could be eliminated by light-soaking treatment. Kelvin probe force microscope (KPFM) measurement proved that a large work function (WF) difference (0.70 eV) between HC-PEDOT:PSS and ZnO is the main reason for the formation of S-shape. White light soaking of the Ag-grid/HC-PEDOT:PSS gradually decreased the WF of HC-PEDOT:PSS from 5.10 to 4.60 eV, leading to a reduced WF difference between HC-PEDOT:PSS and ZnO from 0.70 to 0.38 eV. Such a WF difference decrease was believed to be the working mechanism for the light-soaking effect in this flexible device. Based on this finding, the HC-PEDOT:PSS solution was then modified by doping with polyethylenimine (PEI) and aqueous ammonia. The modified PEDOT:PSS film is characteristic of adjusting WF through varying PEI doping concentrations. By using such a modified PEDOT:PSS layer, light-soaking-free flexible inverted polymer solar cell with a power conversion efficiency of 6.58% was achieved for PTB7-Th:PC71BM cells. The current work provides a useful guideline for interfacial modification for Ag-grid based flexible electrode.


Surgical site infections in day surgery settings.
Because major complications are rare in day surgery, minor morbidities such as surgical site infections, which become evident only after patient discharge, have a major impact on the perceived quality of care. We performed an audit of such infections over a oneyear period. All procedures were performed in a multidisciplinary day surgery setting. There were 155 inguinal hernia repairs, 44 excisions of lipomas, and 27 excisions of pilonidal cysts. All patients received antibiotic prophylaxis according to a rotation protocol. One patient (0.65%) operated on for inguinal hernia developed a deep incisional infection caused by Staphylococcus schleiferi. The patient was treated using selective antibiotics, and complete recovery ensued. One patient (2.27%) who underwent excision of a lipoma revealed developed a deep incisional infection associated with fever (39 degrees C) caused by Staphylococcus aureus. He was treated by daily antibiotics and application of dressings for two weeks and recovered. One patient (3.7%) treated for pilonidal cyst developed a deep incisional surgical site infection. A group A Streptococcus was isolated, and he received an intravenous antibiotic for ten days. In the seven patients treated for pilonidal cyst with the wound left open, swab specimens were taken because of persisting local pain, and the following bacteria were found: Pseudomonas aeruginosa, Escherichia coli, Staphylococcus epidermidis, Proteus vulgaris, and Streptococcus constellatus. No antibiotic therapy was given, and local wound care led to complete recovery. In our day surgery unit, the rates of surgical site infection are similar to those described in the literature. Continuing audit is essential to maintain and improve the quality and standard of the surgery provided. Prevention of surgical site infection is very much the responsibility of the persons working in the operating theater. Therefore, it is vital not only to apply strict regulations concerning everyday management but also to maintain a high degree of motivation among the personnel.


An overview of biological signal processing: non-linear and non-stationary aspects.
The elucidation of biological systems will be one of great keys to scientific advance. However, it is difficult to analyze such biological systems because of their non-linear and non-stationary characteristics. This paper reviews newly developed methods for analysing such non-linear and non-stationary characteristics.


Effects of NH4+ on reflexes in cat spinal cord.
1. In deeply barbiturate-anesthetized animals. NH4+ decreases spinal excitatory synaptic transmission by neuronal depolarization and subsequent block of conduction of action potentials into presynaptic terminals of low-threshold (presumably Ia-) afferents. Because barbiturates by themselves depress excitatory synaptic transmission and may have modified the effects of NH4+, this study examines the effect of NH4+ on excitatory synaptic transmission in the unanesthetized animal. 2. The effects of NH4+ on monosynaptic and polysynaptic excitatory reflexes as well as di- and polysynaptic inhibition were investigated in the spinal cord of the decerebrate and unanesthetized cat in vivo. 3. The monosynaptic excitatory reflex (MSR) elicited by muscle nerve stimulation and polysynaptic excitatory reflexes elicited by muscle (MSR-PSR) or cutaneous nerve stimulation (Cut-PSR) were recorded from the ventral roots L7 or S1. The P-wave was recorded from the cord dorsum. Di- and polysynaptic inhibition was elicited by muscle nerve stimulation and measured as decrease of the MSR. 4. Intravenous infusion of ammonium acetate (AA) decreased MSR and the monosynaptic motoneuron pool excitatory postsynaptic potential (EPSP) recorded from the ventral root (VR-EPSP). Decrease of MSR and VR-EPSP was accompanied by an increase of the intraspinal conduction time in presynaptic terminals. The maximal decrease of the MSR was preceded by a period of transient increase of the MSR and reflex discharges from previously subthreshold VR-EPSPs. 5. The effects of NH4+ on MSR and VR-EPSP are consistent with those in barbiturate-anesthetized animals and suggest that NH4+ also decreases monosynaptic excitation in unanesthetized animals by depolarization and subsequent conduction block for action potentials in presynaptic terminals. 6. Decrease of the MSR was accompanied by a decrease of the P-wave, indicating that NH4+ simultaneously decreases mono- and oligosynaptic excitatory synaptic transmission as well as presynaptic inhibition. 7. Decrease of the MSR was accompanied by increases of MSR-PSR and Cut-PSR and decreases of di- and polysynaptic postsynaptic inhibition. 8. The neuronal circuits underlying MSR-PSR and Cut-PSR include presynaptic inhibition of group I and II afferents as well as postsynaptic inhibition of motoneurons. It is suggested that increases of MSR-PSR and Cut-PSR are contributed to by decreases of pre- and postsynaptic inhibition and neuronal depolarization by NH4+. These effects increase afferent input to motoneurons, permit uncontrolled discharge of motoneurons, and initiate reflex discharges by previously subthreshold excitatory postsynaptic potentials.


Demonstration of cytomegalovirus retinitis by in situ DNA hybridization.
Five cases of the acquired immune deficiency syndrome (AIDS), each exhibiting retinitis, were studied by DNA hybridization technique to detect viral infection. Five involved eyes were obtained at the time of autopsy from three patients who had received no treatment for the retinitis. Retinal biopsy specimens were obtained at the time of surgery from two other patients who developed retinal detachments after being treated with ganciclovir (dihydroxy propoxymethyl guanine). DNA hybridization revealed cytomegalovirus in retina and retinal pigment epithelium in all five specimens from patients who had not been treated with ganciclovir. No hybridization occurred in the two retinal biopsy specimens obtained from the ganciclovir-treated patients. These results suggest that in situ DNA hybridization is a highly specific and easily interpretable means of establishing the tissue diagnosis of viral retinitis.


Multicenter data to improve health for pediatric renal transplant recipients in North America: Complementary approaches of NAPRTCS and IROC.
Kidney transplantation increases life expectancy and improves quality of life for children with end-stage kidney disease, yet sequelae of transplantation and treatment make it difficult for transplant recipients to enjoy health and quality of life similar to their healthy peers. The NAPRTCS network was among the first to use multicenter data to inform improvements in care and outcomes for children with a kidney transplant through observational research. Now, with new technologies and unprecedented access to data, it is possible to create learning health systems as envisioned by the US National Academy of Sciences to seamlessly integrate research and continuous improvement of clinical care. In this review, we present two pre-eminent North American networks focused on using multicenter data to drive improved care and outcomes for children with a kidney transplant. Whereas, for the past 30 years NAPRTCS has focused on discovery of best practices through observational research and clinical trials, the Improving Renal Outcomes Collaborative, established in 2016, engages patients, families, clinicians, and researchers in redesigning the healthcare delivery system to enable practice change and continuous improvement of health outcomes. We discuss the history and past contributions of these networks, as well as current activities, barriers, and potential future solutions to more fully realize the vision of a true learning health system for pediatric kidney transplant recipients.


Search for the rare decays B-->Kl(+)l(-) and B-->K(*)l(+)l(-).
We present results from a search for the flavor-changing neutral current decays B-->Kl(+)l(-) and B-->K(*)l(+)l(-), where l(+)l(-) is either an e(+)e(-) or mu(+)mu(-) pair. The data sample comprises 22.7 x 10(6) Upsilon(4S)-->B(-)B decays collected with the BABAR detector at the PEP-II B Factory. We obtain the 90% C.L. upper limits B(B-->Kl(+)l(-))<0.51 x 10(-6) and B(B-->K(*)l(+)l(-))<3.1 x 10(-6), close to standard model predictions for these branching fractions. We have also obtained limits on the lepton-family-violating decays B-->Ke+/-mu(-/+) and B-->K(*)e(+/-)mu(-/+).


Renal effects of Bunodosoma caissarum crude extract: Prostaglandin and endothelin involvement.
Sea anemones contain a variety of interesting biologically active compounds, including some potent toxins. PLA2 from Bunodosoma caissarum, a sea anemone endemic in the Brazilian southern coast, has shown renal alterations on isolated kidney. The aim of this study was to evaluate the renal and vascular effects of B. caissarum crude extract (BcE) on isolated perfused kidney and arteriolar mesenteric bed, as well the involvement of prostaglandins and endothelin. BcE did not show any effect on arteriolar mesenteric bed, but increased perfusion pressure, renal vascular resistance, urinary flow, glomerular filtration rate and decreased the percentage of sodium tubular transport on isolated perfused kidney. Indomethacin blocked the renal effects induced by BcE and tezosentan only partially blocked these effects. These results demonstrate the effects of BcE on kidney in situ, suggesting the involvement of prostaglandins and endothelin.


Emergence of a drug-dependent human immunodeficiency virus type 1 variant during therapy with the T20 fusion inhibitor.
The fusion inhibitor T20 belongs to a new class of anti-human immunodeficiency virus type 1 (HIV-1) drugs designed to block entry of the virus into the host cell. However, the success of T20 has met with the inevitable emergence of drug-resistant HIV-1 variants. We describe an evolutionary pathway taken by HIV-1 to escape from the selective pressure of T20 in a treated patient. Besides the appearance of T20-resistant variants, we report for the first time the emergence of drug-dependent viruses with mutations in both the HR1 and HR2 domains of envelope glycoprotein 41. We propose a mechanistic model for the dependence of HIV-1 entry on the T20 peptide. The T20-dependent mutant is more prone to undergo the conformational switch that results in the formation of the fusogenic six-helix bundle structure in gp41. A premature switch will generate nonfunctional envelope glycoproteins (dead spikes) on the surface of the virion, and T20 prevents this abortive event by acting as a safety pin that preserves an earlier prefusion conformation.


Gene and metabolite expression dependence on body mass index in human myocardium.
We hypothesized that body mass index (BMI) dependent changes in myocardial gene expression and energy-related metabolites underlie the biphasic association between BMI and mortality (the obesity paradox) in cardiac surgery. We performed transcriptome profiling and measured a panel of 144 metabolites in 53 and 55, respectively, myocardial biopsies from a cohort of sixty-six adult patients undergoing coronary artery bypass grafting (registration: NCT02908009). The initial analysis identified 239 transcripts with biphasic BMI dependence. 120 displayed u-shape and 119 n-shape expression patterns. The identified local minima or maxima peaked at BMI 28-29. Based on these results and to best fit the WHO classification, we grouped the patients into three groups: BMI < 25, 25 ≤ BMI ≤ 32, and BMI > 32. The analysis indicated that protein translation-related pathways were downregulated in 25 ≤ BMI ≤ 32 compared with BMI < 25 patients. Muscle contraction transcripts were upregulated in 25 ≤ BMI ≤ 32 patients, and cholesterol synthesis and innate immunity transcripts were upregulated in the BMI > 32 group. Transcripts involved in translation, muscle contraction and lipid metabolism also formed distinct correlation networks with biphasic dependence on BMI. Metabolite analysis identified acylcarnitines and ribose-5-phosphate increasing in the BMI > 32 group and α-ketoglutarate increasing in the BMI < 25 group. Molecular differences in the myocardium mirror the biphasic relationship between BMI and mortality.


Conformation of the 3'-end of beet necrotic yellow vein benevirus RNA 3 analysed by chemical and enzymatic probing and mutagenesis.
Secondary structure-sensitive chemical and enzymatic probes have been used to produce a model for the folding of the last 68 residues of the 3'-non-coding region of beet necrotic yellow vein benevirus RNA 3. The structure consists of two stem-loops separated by a single-stranded region. RNA 3-derived transcripts were produced containing mutations which either disrupted base pairing in the helices or maintained the helices but with alterations in the base pairing scheme. Other mutants contained substitutions in single-stranded regions (loops or bulged sequences). With a few exceptions all three types of mutation abolished RNA 3 replication in vivo, suggesting that both secondary structure and specific sequences are required for efficient recognition of the 3'-terminal region of RNA 3 by viral RNA-dependent RNA polymerase.


Antiviral activities of tragacanthin polysaccharides on Punta Toro virus infections in mice.
Tragacanthin polysaccharides from Astragalus brachycentrus (AV208) and Astragalus echidnaeformis (AV212) plants, which are devoid of in vitro antiviral activity, were evaluated in a mouse model of Punta Toro virus (PTV) infection. The PTV (a phlebovirus member of the Bunyaviridae family of viruses) is a model for studying the treatment of Rift Valley fever and hantavirus infections. Single intraperitoneal treatments with 12.5-200 mg/kg/day doses of AV212 given 24 h before or 4 and 24 h after virus inoculation protected the majority of mice from mortality. Single treatments with AV208 were ineffective when given 24 h before the virus challenge; however, protection was afforded when treatments were administered at 4 and 24 h following virus inoculation. In a follow-up study, AV208 treatments of 1.6-50 mg/kg/day given 24 h subsequent to virus exposure caused reductions in mortality, liver infection scores, liver and spleen virus titers, and serum transaminases. The polysaccharides did not activate lymphocytes or natural killer cells, nor was interferon induced in treated mice. However, mice pretreated with fumed silica (a macrophage poison) and infected with the PTV were not protected by subsequent administration of AV208 or AV212 at 50 mg/kg, providing evidence that activation of peritoneal macrophages by the polysaccharides affords protection to infected animals. These compounds should be considered for the potential treatment of significant human infections induced by bunyaviruses and hantaviruses.


Childhood craniopharyngioma: hypothalamus-sparing surgery decreases the risk of obesity.
Craniopharyngioma is a brain tumor whose high local recurrence rate has for a long time led to a preference for extensive surgery. Limited surgery minimizing hypothalamic damage may decrease the severe obesity rate at the expense of the need for radiotherapy to complete the treatment. We compared weight gain and local recurrence rates after extensive resection surgery (ERS) and hypothalamus-sparing surgery (HSS). Our observational study compared a historical cohort managed with ERS between 1985 and 2002 to a prospective cohort managed with HSS between 2002 and 2010. The patients were treated in a pediatric teaching hospital in Paris, France. Thirty-seven boys and 23 girls were managed with ERS (median age, 8 years); 38 boys and 27 girls were managed with HSS (median age, 9.3 years). Data were collected before and 6 months to 7 years after surgery. Body mass index (BMI) Z-score was used to assess obesity and the number of surgical procedures to assess local recurrence rate. Mean BMI Z-score before surgery was comparable in the 2 cohorts (0.756 after ERS vs 0.747 after HSS; P = .528). At any time after surgery, mean BMI Z-score was significantly lower after HSS (eg, 1.889 SD vs 2.915 SD, P = .004 at 1 year). At last follow-up, the HSS cohort had a significantly lower prevalence of severe obesity (28% vs 54%, P < .05) and higher prevalence of normal BMI (38% vs 17%, P < .01). Mean number of surgical procedures was not significantly different in the 2 cohorts. Hypothalamus-sparing surgery decreases the occurrence of severe obesity without increasing the local recurrence rate.


Spatio-temporal changes in the speed of canopy development and senescence in temperate China.
China has been suggested to be the country with the largest vegetation greenness over the last four decades. In this study, we investigated the change in the speed of canopy development and senescence as well as its linkage with climate at monthly scale across temperate China, using satellite-derived Normalized Difference Vegetation Index (NDVI) data from 1982 to 2015. A significant increase in mean monthly NDVI occurred across all growing-season months except June and November, but this greening trend was mainly contributed by the faster speed of canopy development in April and the slower speed of canopy senescence in October. The average of VNDVI (the difference in NDVI between 2 consecutive months) over temperate China is significantly increased only in April (7.75 × 10-4 year-1 , p < .05) and October (4.98 × 10-4 year-1 , p < .05). In contrast, VNDVI in November is significantly decreased, indicating an increasing trend in the magnitude of leaf fall in the last month of the growing season due to both increase in season maximum greenness and slower canopy senescence in October. We also found clear seasonal differences in the correlations between VNDVI and climatic factors, especially temperature. In April and October, the correlations between VNDVI and temperature were generally positive, while they were negative in June. VNDVI in spring (early growing season) and summer (middle growing season) was also positively correlated with precipitation in semiarid regions. Such seasonally distinct climatic controls on VNDVI should be considered in modelling vegetation responses to climate change. Overall, our findings can help quantify the contribution of different climatic drivers on the shifts in canopy development and senescence and better elucidate the change in vegetation greenness under future climate change.


Language laterality in English/Welsh bilinguals: language-acquisitional and language-specific factors in the development of lateralisation.
In order to test the hypothesis that monolinguals differ from bilinguals in their pattern of language lateralisation and to examine the relative merits of language-acquisitional versus language-specific factors, two experiments involving divided screen presentation of two languages were conducted using Welsh/English speaking participants. In the first experiment 80 monolingual teenagers were compared to 80 bilingual teenagers on a tachistoscopic "visual half-field" test of Welsh and English nouns and verbs. ANOVA revealed a greater left hemisphere advantage for Welsh-English bilinguals as compared to English monolinguals. Thus, in contrast to previous studies, in our bilinguals there was evidence of greater left hemisphere involvement in the processing of language. In the second experiment, four separate groups of 40 teenagers, varying in the age and manner of acquisition of their languages, were compared on the same test of Welsh and English words. These groups can be viewed as graded from the early to late bilinguals. ANOVA revealed a greater left hemisphere advantage when processing Welsh as compared to English words for all four groups. However no significant difference was observed between the four groups in respect of laterality for Welsh and English, indicating an equally greater left hemisphere bias for all four groups when processing Welsh words. We discuss these results in terms of a language-specific effect and suggest the specific orthography of the Welsh language (for individually presented nouns and verbs) promotes a left hemisphere advantage over and above language-acquisitional factors.


The Role of n-3 Fatty Acids in Cardiovascular Disease: Back to the Future.
Cardiovascular disease (CVD) remains the major cause of death and disability worldwide, and residual risk after implementing all current therapies is still high. In this context, the latest (2016) European Cardiology Society/European Atherosclerosis Society guidelines recommend that triglyceride (TG)-lowering drugs should be used in high-risk patients with TGs levels >2.3 mmol/L (200 mg/dL), after lifestyle measures fail to lower them. After several neutral CVD outcome trials with n-3 fatty acids, the Reduction of Cardiovascular Events with EPA-Intervention Trial met its primary end point, that is, among patients with elevated TGs levels despite the use of statins, the risk of ischemic events, including cardiovascular death, was significantly lower in those who received 4 g of icosapent ethyl daily. In this review, we comment on the findings of previous and recently published randomized controlled CVD outcome trials assessing n-3 fatty acids supplementation. Both efficacy and safety, as well as future perspectives, are discussed.


1,2-dithiin annelated with bicyclo[2.2.2]octene frameworks. One-electron and two-electron oxidations and formation of a novel 2,3,5,6-tetrathiabicyclo[2.2.2]oct-7-ene radical cation with remarkable stability owing to a strong transannular interaction.
A stable derivative of 1,2-dithiin annelated with bicyclo[2.2.2]octene frameworks 4 was synthesized as red crystals by the reaction of a dilithiated dimer of bicyclo[2.2.2]octene with elemental sulfur in 59% yield. The cyclic voltammetry of 4 in CH(2)Cl(2) at -78 degrees C showed two reversible oxidation waves at E(1/2) +0.18 V and +0.72 V versus Fc/Fc(+), indicating that the radical cation and dication of 4 are stable under these conditions. Upon chemical one-electron oxidation of 4 in a rather low concentration (4.0 x 10(-4) M) with a 1.5 equiv of SbCl(5) in CH(2)Cl(2), a radical cation 4.+ was formed, whose spin distribution was determined by ESR spectroscopy and by the results of theoretical calculations (UB3LYP/6-31G). The electronic absorption spectrum of 4.+ in CH(2)Cl(2) exhibited a maximum absorption at 428 nm (epsilon = 2.3 x 10(3)), which was hypsochromically shifted from that of neutral 4 (469 nm). When the radical cation 4.+ was produced in higher concentration (0.06 M) in CH(2)Cl(2), a disproportionation was found to take place to give a SbCl(6)(-) salt of remarkably stable radical cation 5.+ having a novel 2,3,5,6-tetrathiabicyclo[2.2.2]oct-7-ene structure. In the X-ray structure of 5.+SbCl(6)(-), the transannular distance (2.794(3) A) between the sulfur atoms was found to be less than the sum of the van der Waals radii of a sulfur atom (3.70 A), suggesting the existence of a bonding interaction between the two disulfide linkages. The theoretical calculations (UB3LYP/6-31G) suggested that this transannular interaction could be described as the resonance between the limiting structures, each of them having a two-center three-electron bond between two sulfur atoms belonging to two different disulfide linkages: thus, both the spin and positive charge are equally delocalized to the four sulfur atoms, causing a great stabilization of 5.+. On the other hand, the 1,2-dithiin radical cation 4.+ was found to readily react with triplet oxygen with subsequent rearrangement to give the 1,2-dithiolium derivative 6+ having a carboxyl group. Finally, the reaction of 4 with an excess amount of SbF(5) gave the corresponding dication 4(2+), which was found to be a 6pi aromatic system on the basis of the results of NMR measurement and theoretical calculations.


Phase and frequency coordination between neuron firing as an integrative mechanism of human CNS self-organization.
It was shown by surface electromyography (sEMG) from spinal cord injury and Parkinson's disease patients that FF-type motor units fire repeatedly at a rate of 10 Hz and FR-type motor units with approximately 3 action potentials per impulse train at 4 to 5 Hz. Such oscillatory firing was compared with the firing of alpha1, alpha2, and alpha3-motoneurons, innervating FF FR, and S-type muscle fibres, which fire oscillatory approximately at 10 Hz, 4 to 7 Hz and 1 Hz, as measured earlier with the single nerve-fibre action potential recording method from motoneuron axons. Repeated firings with impulse trains consisted of 1, 2-5, and 20-50 action potentials per impulse train respectively. Oscillatory firing of motoneurons has now been demonstrated in humans with two methods. In spinal cord injury patients it was observed that the FF-type motor units, firing normally at 10 Hz, fired for higher activations also at approximately 20 Hz. A motor program burst was generated by recruiting for the burst time motor units to fire oscillatory. It was further shown that FF-type motor units fired with relative phase and frequency coordination but non-synchronously. The coordination was between motor units firing of the same muscle and between muscles of the right and left arm. With the single-nerve fibre action potential recording method it was found earlier that alpha and gamma-motoneurons and muscle spindle afferents fired in relative coordination. There is indication that relative phase and frequency coordination is an integrative mechanism for the self-organization of the neuronal networks of the human central nervous system (CNS).


Phorbol diester treatment promotes enhanced adenylate cyclase activity in frog erythrocytes.
Incubation of intact frog erythrocytes with 12-O-tetradecanoyl phorbol-13-acetate (TPA), a tumor-promoting phorbol diester which activates protein kinase C, results in an approximate two- to threefold increase in subsequently tested beta-adrenergic agonist-stimulated adenylate cyclase activity. This increase is due to an elevation in the Vmax of the enzyme rather than to a change in affinity for the agonist. TPA treatment of frog erythrocytes does not alter the affinity (KD) or the binding capacity (Bmax) for the beta-adrenergic antagonist [125I]cyanopindolol. In addition, agonist/[125I]cyanopindolol competition curves are not affected by TPA pretreatment nor is their sensitivity to guanine nucleotides. Incubation of frog erythrocyte membranes alone with TPA does not promote sensitization or activation of adenylate cyclase activity. Pretreatment of intact frog erythrocytes with TPA also produces approximately two- to threefold increases in basal, guanine nucleotide-, prostaglandin E1-, forskolin-, NaF-, and MnCl2-stimulated adenylate cyclase activities in frog erythrocyte membranes. This enhancement of adenylate cyclase activity by TPA is induced rapidly (t1/2 approximately equal to 5 min) and with an EC50 of about 10(-7) to 10(-6) M. Other tumor-promoting phorbol diesters or phorbol diester-like compounds including 4 beta-phorbol 12,13-dibutyrate, 4 beta-phorbol 12,13-didecanoate, and mezerein are effective in promoting enhanced adenylate cyclase activity. In contrast, phorbols such as 4 beta-phorbol, 4 alpha-phorbol 12,13-didecanoate, and 4-O-methylphorbol 12-myristate 13-acetate, which are inactive in tumor promotion and which do not activate protein kinase C, do not affect frog erythrocyte adenylate cyclase activity. These data are suggestive of a protein kinase C-mediated phosphorylation of one of the adenylate cyclase components that is distal to the receptor, i.e., the nucleotide regulatory and/or catalytic components.


A case of histiocytic sarcoma presenting with primary bone marrow involvement.
Histiocytic sarcoma (HS) is a very rare neoplasm that often shows an aggressive clinical course and systemic symptoms, such as fever, weight loss, adenopathy, hepatosplenomegaly and pancytopenia. It may present as localized or disseminated disease. We describe here a 63-yr-old male who manifested systemic symptoms, including fever, weight loss and generalized weakness. Abdominal and chest computed tomography failed to show specific findings, but there was suspicion of multiple bony changes at the lumbar spine. Fusion whole body positron emission tomography, bone scan and lumbar spine magnetic resonance imaging showed multiple bone lesions, suggesting a malignancy involving the bone marrow (BM). Several BM and bone biopsies were inconclusive for diagnosis. Necropsy showed replacement of the BM by a diffuse proliferation of neoplastic cells with markedly increased cellularity (95%). The neoplastic cells were positive for lysozyme and CD68, but negative for T- and B-cell lineage markers, and megakaryocytic, epithelial, muscular and melanocytic markers. Morphologic findings also distinguished it from other dendritic cell neoplasms.


Characterization of the secreted antigens of Mycobacterium bovis BCG: comparison of the 46-kilodalton dimeric protein with proteins MPB64 and MPB70.
Western blot analysis showed that the 46-kilodalton (kDa) dimeric protein antigen secreted in large amounts by some daughter strains of Mycobacterium bovis BCG corresponded to protein MPB70 present in long-term culture filtrates of the Japanese substrain. The 46/23-kDa antigen is the most abundant protein in supernatant from a 5-day culture but is masked by leaked products in old culture supernatants. No similarities were found between the 46-kDa protein and MPB64, a protein with the same strain distribution, or with the antigen of similar molecular mass recognized by monoclonal antibody SA1.D2D.


Interaction of the Belousov-Zhabotinsky Reaction with Phospholipid Engineered Membranes.
Compartmentalized in liposome arrays, the Belousov-Zhabotinsky (BZ) oscillatory reaction might represent a good model for biochemical networks. In order to engineer such liposomes, we used small-angle X-ray scattering (SAXS) to study the effect of individual BZ reactant as well as of the entire BZ mixture on the structural properties of lipid layer(s) formed by 1,2-dimyristoyl-sn-glycero-3-phosphocholine (DMPC) phospholipids in aqueous media. These properties were compared with those of lipid layers doped with myristic acid (Myr-A), sodium tetradecyl sulfate (STS), and cholesterol (CHOL). In parallel, the effect on the BZ reaction exerted by doped DMPC liposomes was investigated by UV-vis spectroscopy, followed by image analysis of the recorded time series. SAXS experiments showed that chemical species involved in the BZ reaction bring small changes to the internal structure of DMPC bilayers. However, ferroin can reduce the liposome lamellarity by being adsorbed on the surface of lipid layers. Also, the presence of charged dopants such as STS and TA tends to reduce the lamellarity of liposomes, while CHOL brings marked changes in the BZ system due to chemical reaction with oxidant species. In particular, an increase of the oscillation frequency is observed when the BZ reaction is carried out in the presence of CHOL-DMPC liposomes. For this behavior, a possible explanation supported by numerical simulations is bromination of CHOL double bonds by BZ intermediates.


Genetic and Life Style Risk Factors for Recurrent Non-alcoholic Fatty Liver Disease Following Liver Transplantation.
Recurrent or de novo non-alcoholic fatty liver disease (NAFLD)/non-alcoholic steatohepatitis (NASH) following liver transplantation (LT) is a frequent event being increasingly recognized over the last decade, but the influence of recurrent NASH on graft and patient outcomes is not yet established. Taking into consideration the long term survival of liver transplanted patients and long term complications with associated morbidity and mortality, it is important to define and minimize risk factors for recurrent NAFLD/NASH. Metabolic syndrome, obesity, dyslipidemia, diabetes mellitus are life style risk factors that can be potentially modified by various interventions and thus, decrease the risk of recurrent NAFLD/NASH. On the other hand, genetic factors like recipient and/or donor PNPLA3, TM6SF2, GCKR, MBOAT7 or ADIPOQ gene polymorphisms proved to be risk factors for recurrent NASH. Personalized interventions to influence the different metabolic disorders occurring after LT in order to minimize the risks, as well as genetic screening of donors and recipients should be performed pre-LT in order to achieve diagnosis and treatment as early as possible.


Steroid recognition by chloramphenicol acetyltransferase: engineering and structural analysis of a high affinity fusidic acid binding site.
The antibiotic fusidic acid and certain closely related steroidal compounds are potent competitive inhibitors of the type I variant of chloramphenicol acetyltransferase (CATI). In the absence of crystallographic data for CATI, the structural determinants of steroid binding were identified by (1) construction in vitro of genes encoding chimaeric enzymes containing segments of CATI and the related type III variant (CATIII) and (2) site-directed mutagenesis of the gene encoding CATIII, followed by kinetic characterisation of the substituted variants. Replacement of four residues of CATIII (Gln92, Asn146, Tyr168 and Ile172) by their equivalents from CATI yields an enzyme variant that is susceptible to competitive inhibition by fusidate with respect to chloramphenicol (Ki = 5.4 microM). The structure of the complex of fusidate and the Q92C/N146F/Y168F/I172V variant, determined at 2.2 A resolution by X-ray crystallography, reveals the inhibitor bound deep within the chloramphenicol binding site and in close proximity to the side-chain of His195, an essential catalytic residue. The aromatic side-chain of Phe146 provides a critical hydrophobic surface which interacts with non-polar substituents of the steroid. The remaining three substitutions act in concert both to maintain the appropriate orientation of Phe 146 and via additional interactions with the bound inhibitor. The substitution of Gln92 by Cys eliminates a critical hydrogen bond interaction which constrains a surface loop (residues 137 to 142) of wild-type CATIII which must move in order for fusidate to bind to the enzyme. Only two hydrogen bonds are observed in the CAT-fusidate complex, involving the 3-alpha-hydroxyl of the A-ring and both hydroxyl of Tyr25 and NE2 of His195, both of which are also involved in hydrogen bonds with substrate in the CATIII-chloramphenicol complex. In the acetyl transfer reaction catalysed by CAT, NE2, of His195 serves as a general base in the abstraction of a proton from the 3-hydroxyl of chloramphenicol as the first chemical step in catalysis. The structure of the CAT-inhibitor complex suggests that deprotonation of the 3-alpha-hydroxyl of bound fusidate by this mechanism could produce an oxyanion nucleophile analogous to that seen with chloramphenicol, but one which is incorrectly positioned to attack the thioester carbonyl of acetyl-CoA, accounting for the observed failure of CAT to acetylate fusidate.


Structural and functional analyses of the optic nerve and lateral geniculate nucleus in glaucoma.
To analyze the correlation between structural characteristics of intraorbital optic nerve (ION) and lateral geniculate nucleus (LGN) measured by 3-Tesla magnetic resonance imaging (3T MRI), and the severity of glaucomatous damage. In this cross-sectional study, 41 glaucoma patients and 12 age- and sex-matched controls underwent standard automated perimetry (SAP) and frequency doubling technology (FDT) as functional evaluation; optic disc stereophotograph, spectral-domain optical coherence tomography (OCT) and confocal scanning laser tomography as ocular structural evaluation; and 3T MRI. Structure-structure and structure-function correlation were performed using bootstrap resampling method for clustered data. The ION mean diameter and cross-sectional area were different between glaucoma and control groups at 5mm and 10mm (all, p≤0.011) from the globe, but not at 15mm (both, p≥0.067). LGN height was significantly lower in glaucoma group (p = 0.005). OCT rim area and functional parameters (SAP and FDT) correlated significantly with all ION segments, showing stronger correlations at 10 and 15 mm. ION parameters at 10 and 15 mm presented mild-to-moderate correlation with OCT peripapillary nerve fiber layer thickness, and ION at 15mm had mild association with the neuroretinal rim area on stereophotographs. Although LGN height was significantly smaller in glaucoma group (p = 0.005), LGN parameters were not associated with any ocular structural or functional parameter. Assessment of central and peripheral nervous systems using 3T MRI confirmed that glaucoma patients had smaller ION dimensions and LGN height compared to the control group. In general, ION dimensions presented mild to moderate correlations with functional and ocular structural parameters. Although ION had significant correlations at any distance from the eye, the ION distal locations correlated better with OCT results and functional parameters. However, LGN parameters were not associated with functional or ocular structural parameters.


Amino acids stimulate translation initiation and protein synthesis through an Akt-independent pathway in human skeletal muscle.
Studies in vitro as well as in vivo in rodents have suggested that amino acids (AA) not only serve as substrates for protein synthesis, but also as nutrient signals to enhance mRNA translation and protein synthesis in skeletal muscle. However, the physiological relevance of these findings to normal humans is uncertain. To examine whether AA regulate the protein synthetic apparatus in human skeletal muscle, we infused an AA mixture (10% Travesol) systemically into 10 young healthy male volunteers for 6 h. Forearm muscle protein synthesis and degradation (phenylalanine tracer method) and the phosphorylation of protein kinase B (or Akt), eukaryotic initiation factor 4E-binding protein 1, and ribosomal protein S6 kinase (p70(S6K)) in vastus lateralis muscle were measured before and after AA infusion. We also examined whether AA affect urinary nitrogen excretion and whole body protein turnover. Postabsorptively all subjects had negative forearm phenylalanine balances. AA infusion significantly improved the net phenylalanine balance at both 3 h (P < 0.002) and 6 h (P < 0.02). This improvement in phenylalanine balance was solely from increased protein synthesis (P = 0.02 at 3 h and P < 0.003 at 6 h), as protein degradation was not changed. AA also significantly decreased whole body phenylalanine flux (P < 0.004). AA did not activate Akt phosphorylation at Ser(473), but significantly increased the phosphorylation of both eukaryotic initiation factor 4E-binding protein 1 (P < 0.04) and p70(S6K) (P < 0.001). We conclude that AA act directly as nutrient signals to stimulate protein synthesis through Akt-independent activation of the protein synthetic apparatus in human skeletal muscle.


The cells of origin of the spinohypothalamic tract in cats.
Various cutaneous and visceral stimuli alter the discharge rates of neurons in the hypothalamus. Changes in the activity of hypothalamic neurons are thought to play important roles in eliciting neuroendocrine, autonomic, and affective responses to somatosensory and visceral stimuli. Information from peripheral structures has been considered generally to reach the hypothalamus via multisynaptic ascending pathways. Recently, a direct projection from the spinal cord to the hypothalamus was demonstrated in rats. The goal of this study was to determine whether a similar projection exists in cats. Either wheat germ agglutinin conjugated to horseradish peroxidase, a mixture of this tracer and the B subunit of cholera toxin conjugated to horseradish peroxidase, or fast blue was injected into the hypothalamus of cats. Injections were centered in the hypothalamus in seven cats and did not spread to the thalamus, zona incerta or midbrain. After these injections, retrogradely labeled neurons were observed bilaterally in each of the 17 spinal segments that were examined. A total of approximately 400-500 labeled neurons was observed in alternate sections through these segments in the most effective cases. Roughly 70% of the labeled neurons were located contralaterally. Labeled neurons were found predominantly in the deep dorsal horn, the intermediate zone/ventral horn and in the area surrounding the central canal. A few were also noted in the superficial dorsal horn. The first and second sacral segments contained the largest numbers of retrogradely labeled neurons in the spinal cord. The number of spinohypothalamic tract neurons observed in this study in cats was roughly an order of magnitude smaller than that previously reported for rats. This finding suggested either that the spinohypothalamic tract is relatively small in cats or that our tracing techniques did not label many spinohypothalamic tract neurons in cats. To test the sensitivity of one of our tracing techniques, control injections of wheat germ agglutinin conjugated to horseradish peroxidase that filled the ventrobasal thalamus were made in two cats. In both cases, thousands of spinal cord neurons were labeled. In summary, our results indicate that a spinohypothalamic tract exists in cats. However, our findings also suggest that the total number of spinohypothalamic tract neurons in cats may be an order of magnitude smaller than it is in rats.


CytoSorb® hemoadsorption and mechanical circulatory support in a newborn with refractory shock after congenital heart surgery.
Refractory cardiogenic and vasoplegic shock after congenital heart surgery is a threatening condition leading to high morbidity and mortality. Control of hemodynamic and inflammatory response is fundamental in medical strategy. We report the case of a newborn with cardiogenic and vasoplegic shock secondary to cardiopulmonary bypass for atrioseptostomy and prostaglandin treatment in the context of hypoplastic left heart syndrome, successfully treated with a combination of mechanical circulatory support and cytokine hemoadsorption column (CytoSorb®). Vasopressor support was weaned during the time of treatment without multiple organ failure occurrence. The use of CytoSorb® cartridge inserted in an extra-corporeal circuit even in a newborn is easy and feasible, as long as some precautions are considered. Routine monitoring of drugs levels is mandatory during the use of the cartridge and immediately after.


In the Spotlight: BioInstrumentation.
This paper discussed physiological measurement instrumentation for ambulatory/wearable physiological monitoring, cardio-pulmonary monitoring, activity monitoring, and biochemical monitoring. Future aspects also were discussed.


A serratial protease causes vascular permeability reaction by activation of the Hageman factor-dependent pathway in guinea pigs.
The 56-kilodalton (56K) protease isolated from a culture filtrate of Serratia marcescens caused vascular permeability enhancement followed by edema formation when injected into guinea pig peripheral corneas and subconjunctival space or skin. The character and the mechanism of permeability enhancement were analyzed in vivo. The enhancement was maximum at 5 to 10 min. The permeability reaction increased exponentially by the amount of enzyme used. The enhancement of permeability induced by the 56K protease was not affected by treatment with an antihistamine but was greatly augmented by simultaneous injection of a kinin potentiator, Glu-Trp-Pro-Arg-Pro-Gln-Ile-Pro-Pro-OH (SQ 20,881). Furthermore, the permeability activity of the protease, but not the amidolytic activity, was inhibited by soybean trypsin inhibitor, a well-known inhibitor of plasma kallikrein, as well as by corn trypsin inhibitor, the best inhibitor of activated Hageman factor. Results of these in vivo studies indicate that the permeability-enhancing reaction induced by the 56K protease is caused by activation of the Hageman factor-dependent pathway in the tissue. The permeability-increasing activity of the 56K protease was parallel with the enzyme activity. Serratial lipopolysaccharide did not produce a permeability enhancement reaction within 30 min when injected into guinea pig skin. These results are consistent with the results of recent in vitro experiments in which activation of the purified Hageman factor but not of prekallikrein by the 56K protease was elucidated (Matsumoto et al., J. Biochem. (Tokyo) 96:739-749, 1984). Thus, the molecular mechanism described above appears to be operative in the pathogenesis of corneal edema and chemosis, which is induced by S. marcescens, in addition to the direct tissue destruction by the protease.


Hydroxychloroquine Blood Levels Predict Hydroxychloroquine Retinopathy.
In 2016, the American Academy of Ophthalmology (AAO) changed the recommended daily dose of hydroxychloroquine (HCQ) from 6.5 mg/kg to <5 mg/kg. However, it is not clear that the lower prescribed dose of HCQ will have the same efficacy for systemic lupus erythematosus (SLE) activity or the same role in protecting against cardiovascular risk factors and thrombosis. This study was undertaken to address the frequency of HCQ retinopathy and the role of HCQ blood levels in identifying those individuals who are at a greater future risk of retinopathy. HCQ blood levels in 537 patients with SLE from a large clinical cohort were repeatedly measured, and patients were tested for HCQ retinopathy. We assessed the risk of retinopathy according to clinical characteristics and blood levels of HCQ. The overall frequency of retinopathy was 4.3% (23 of 537 patients). There was a 1% risk of retinopathy in the first 5 years of HCQ treatment, 1.8% from 6 to 10 years, 3.3% from 11 to 15 years, 11.5% from 16 to 20 years, and 8.0% after 21 years of use. We found that older age (P < 0.0001), higher body mass index (P for trend = 0.0160), and longer duration of HCQ intake (P = 0.0024 and P for trend = 0.0006) were associated with a higher risk of HCQ toxicity. Higher blood levels of HCQ predicted later HCQ retinopathy (P = 0.0124 and P = 0.0340 for mean and maximum HCQ blood levels, respectively). Our data prove the utility of assessing blood levels of HCQ in the prediction of retinopathy. This would allow clinicians to either decrease the dose or increase monitoring in those patients with high HCQ blood levels.


Isolation and analysis of the molecular epidemiology and zoonotic significance of Mycobacterium tuberculosis in domestic and wildlife ruminants from three states in India.
The majority of tuberculosis cases in ruminants are caused by Mycobacterium bovis (MB). However, in this study, the authors reported the isolation of Mycobacterium tuberculosis (MT) from bovine milk, nasal swabs and post-mortem tissue samples (n = 841) collected from cattle and buffaloes in the states of Telangana, Maharashtra and Gujarat in India in the period from 2010 to 2015. The isolates (n = 7) were confirmed as Mycobacterium due to their growth characteristics and colony morphology in a commercial liquid medium Mycobacterial Growth Indicator Tube (MGIT)™ employing the BD BACTEC™ MGIT™ 960 system and the Löwenstein-Jensen (LJ) medium supplemented with glycerol but not with sodium pyruvate, and BD-DIFCO™ Middlebrook 7H10 agar containing oleic albumin dextrose catalase (OADC). These isolates were initially identified as members of the M. tuberculosis complex (MTC) using a commercial nested polymerase chain reaction (PCR) kit based on the IS6110 MTC specific nucleotide sequence. The isolates were confirmed as MT using three commercial line probe assay kits, were further genotyped, and the spoligotypes identified were of East African Indian (EAI) 3_IND, EAI5, Central-Asian (CAS) 1_DELHI, U and T1 lineages. Two MT isolates from one antelope (Antilope cervipara) andone gazelle (Gazella bennettii) from Gujarat, which were identified previously, were spoligotyped during this study and identified as belonging to EAI3_IND and EAI5 lineages, respectively. The epidemiological significance and zoonotic implications of regional presence and documentation of the same or two differents poligotypes in different species within the family Bovidae as well as humans is discussed.


Loss of gastric interstitial cells of Cajal in patients with hereditary transthyretin amyloidosis.
Hereditary transthyretin (TTR) amyloidosis is a systemic neuropathic disorder caused by TTR gene mutations. Gastrointestinal complications are common and the underlying mechanisms remain unclear. The interstitial cells of Cajal (ICC) function as pacemaker cells in the gastrointestinal tract and are important for gastrointestinal motility. The aim of this study was to investigate the densities of gastric ICC and nerves in patients with TTR amyloidosis compared to non-amyloidosis controls. Antral wall autopsy specimens from 11 Japanese ATTR V30M patients and 10 controls were analyzed with immunohistochemistry and computerized analysis. Antibodies to c-Kit and TMEM16A were used to assess ICC and an antibody to PGP 9.5 was used to assess nervous tissue. The study was approved by a Japanese ethical committee. The densities of c-Kit-immunoreactive (IR) ICC were significantly lower in the circular and longitudinal muscle layers of patients compared to controls (p = 0.004 for both). Equivalent results were found for TMEM16A-IR ICC. There were no significant differences in PGP 9.5-IR cells in the circular or longitudinal muscle layers between patients and controls (p = 0.173 and 0.099, respectively). A loss of gastrointestinal ICC may be an important factor for the digestive disturbances in hereditary TTR amyloidosis.


Strategy of topical vaccination with nanoparticles.
Liposomes in the nanosize range have been recognized as a versatile drug delivery system of both hydrophilic and lipophilic molecules. In order to develop a liposome-based topical vaccination strategy, five different types of liposomes were tested as a putative vaccine delivery system on pig ear skin. The investigated liposomes mainly varied in size, lipid composition, and surface charge. Using hydrophilic and hydrophobic fluorescent dyes as model drugs, penetration behavior was studied by means of confocal laser scanning microscopy of intact skin and histological sections, respectively. Follicular penetration of the liposomes was measured in comparison to a standard, nonliposomal formulation at different time points. Dependent on time but independent of their different characters, the liposomes showed a significantly higher penetration depth into the hair follicles compared to the standard formulation. The standard formulation reached a relative penetration depth of 30% of the full hair follicle length after seven days, whereas amphoteric and cationic liposomes had reached approximately 70%. Penetration depth of negatively charged liposomes did not exceed 50% of the total follicle length. The fluorescence dyes were mainly detected in the hair follicle; only a small amount of dye was found in the upper parts of the epidermis.


Telomeric localization of the vertebrate-type hexamer repeat, (TTAGGG)n, in the wedgeshell clam Donax trunculus and other marine invertebrate genomes.
The hexamer repeat sequence (TTAGGG)(n), found at the ends of all vertebrate chromosomes, was previously identified as the main building element of one member of a HindIII satellite DNA family characterized in the genome of the bivalve mollusc Donax trunculus. It was also found in 22 perfect tandem repeats in a cloned junction region juxtaposed to the proper satellite sequence, from which the DNA tract encompassing the clustered tandem copies was excised and subcloned. Here, the chromosomal distribution of (TTAGGG)(n) sequences in the Donax was studied by the sensitivity to Bal31 exonuclease digestion, fluorescence in situ hybridization (FISH) on metaphase chromosomes and rotating-field gel electrophoresis. To verify the occurrence of the hexamer repeat in the genomes of taxonomically related molluscs and other marine invertebrates, genomic DNA from the mussel Mytilus galloprovincialis and the echinoderm Holothuria tubulosa was also analyzed. The kinetics of Bal31 hydrolysis of high molecular mass DNA from the three marine invertebrates revealed a marked decrease over time of the hybridization with the cloned (TTAGGG)(22) sequence, concomitantly with a progressive shortening of the positively reacting DNA fragments. This revealed a marked susceptibility to exonuclease consistent with terminal positioning on the respective chromosomal DNAs. In full agreement, FISH results with the (TTAGGG)(22) probe showed that the repeat appears located in telomeric regions in all chromosomes of both bivalve molluscs. The presence of (TTAGGG)(n) repeat tracts in marine invertebrate telomeres points to its wider distribution among eukaryotic organisms and suggests an ancestry older than originally presumed from its vertebrate distinctiveness.


Different phenotypes of intermittent and persistent respiratory allergy in Zagreb, Croatia.
Allergic rhinitis (AR) is a major chronic respiratory disease because of its prevalence, its impacts on the quality of life, economic burden and links with asthma. A significant relationship between the severity of rhinitis and prevalence of asthma in allergic patients was found both in patients suffering from seasonal rhinitis and in those suffering from perennial-allergic rhinitis (PAR) and asthma. The aim of the study was to investigate allergy phenotypes in patients with intermittent allergic rhinitis (IAR) and persistent allergic rhinitis (PAR) in residents of Zagreb, and to determine if there were any other differences in the clinical (in vivo) and diagnostic (in vitro) presentations of the phenotype of subjects suffering from different types of rhinitis. 205 subjects were divided into 2 groups, 102 with IAR sensitized to ragweed pollen (Ambrosia elatior) and 103 with PAR sensitized to house dust mites (Dermatophagoides pteronyssinus and Dermatophagoides farinae). The diagnosis was based on a detailed medical history, examination and diagnostic tests (spirometry, reversibility after appliance of salbutamol, skin prick test, and total IgE). Rhinitis symptoms in subjects with IAR were significantly different than in those with PAR. Sneezing and a runny, itchy nose were frequently present in the IAR group, but not in the PAR group. Prevalence of the coexistence of asthma was higher in the PAR group (35:57%), as well as more severe clinical phenotype of asthma. The covariation of sensitization was similar (70:74%). The most frequent sensitization in the IAR group was found to house dust mite (38%), and in the PAR group to animal dander (40%). The clinical presentation of the IAR was different from that of the PAR. The prevalence of coexisting asthma was significantly higher in the PAR group. The covariation of sensitization was similar.


Folate-deficient human lymphoblasts: changes in de novo purine and pyrimidine synthesis and phosphoribosylpyrophosphate.
Peripheral blood lymphocytes from healthy volunteers cultured with phytohaemagglutinin in a folate-deficient medium exhibit megaloblastic maturation and reduced cellular folate content. For these cells, changes in de novo purine and pyrimidine synthesis and cellular phosphoribosylpyrophosphate (PRPP) have been determined. Mitogen-stimulated cells cultured with undialyzed pooled human serum (PHS) exhibit undetectable de novo purine synthesis, a three-fold increase in PRPP content and augmented de novo pyrimidine synthesis; these changes are independent of cellular folate status. Folate-replete cells cultured with PHS which was dialyzed to reduce purine compounds concentrations show markedly increased de novo purine synthesis. The PRPP content and pyrimidine synthesis rates of these cells are similar to those of folate-replete cells cultured with undialyzed PHS. Folate-deficient cells cultured in dialyzed PHS show a 10-fold reduction in purine synthesis and a corresponding increase in PRPP levels. Pyrimidine synthesis was moderately reduced. The purine bases hypoxanthine and adenine markedly reduced the augmented purine synthesis of folate-replete cells or the increased PRPP content of folate-deficient cells cultured with dialyzed PHS. These findings suggest that cellular folate status is critical for de novo purine synthesis only when coupled with purine bases restriction and that the latter are efficient regulators of de novo purine synthesis.


Does oxygen help dyspnea in patients with cancer?
Dyspnea in patients with advanced cancer is a common symptom that is difficult to treat. This study investigated whether oxygen helps to relieve rest dyspnea in patients with advanced cancer. In a single-blind controlled trial, oxygen and air were administered in random order to hospice patients reporting dyspnea at rest. Measurements of arterial oxygen saturation, lung function, and dyspnea (using a visual analogue scale [VAS] and Borg score) were made before and after each gas had been given for 15 min. Data from 38 patients were used: analysis of variance revealed that mean VAS levels during baseline conditions, breathing room air (59 mm), were significantly reduced after administration of either air (48 mm; p < 0.001) or oxygen (45 mm; p < 0.001); there was no significant difference for the mean VAS scores between oxygen and air administration. There was no statistically significant order of treatment effect. There was no difference in the response to oxygen or air in patients with a history of cardiopulmonary disease. The improvement in dyspnea with oxygen could not be predicted from a subject's initial level of hypoxia. Results suggested that benzodiazepines may potentiate the effect of oxygen. The overall conclusion is that oxygen and air can have a significant effect in reducing dyspnea at rest in patients with advanced cancer.


Utility of paraffin section immunohistochemistry for C-KIT (CD117) in the differential diagnosis of systemic mast cell disease involving the bone marrow.
Systemic mast cell disease is characterized by an abnormal infiltration of mast cells involving several parenchymal organs and the bone marrow. Its spectrum of clinical and histologic presentation is highly variable and is not necessarily correlated with prognosis. Mast cell disorders presenting as atypical infiltrates in the bone marrow may simulate or be associated with other hematolymphoid malignancies, from which they must be distinguished. The paucity of reliable histochemical and immunohistochemical markers for the detection of mast cells in paraffin sections further confounds this diagnosis. The authors have employed immunohistochemistry for the C-KIT encoded tyrosine kinase receptor protein, CD117, for detection of mast cells on paraffin sections of 89 bone marrow specimens including systemic mast cell disease and other disorders. CD117 staining was found in all cases of mast cell disorders (seven of seven), and in one case of chronic myelogenous leukemia in blast crisis. None of the other myeloid disorders tested (0 of 16), or any of the cases of Hodgkin's disease (0 of 12), B-cell lymphomas (0 of 32), T-cell lymphomas (0 of 3), or histiocytic proliferations (0 of 3) showed staining for CD117. CD117 expression is effective in the separation of mast cell disease from disorders that may simulate it histologically.


The role of carvedilol in the management of portal hypertension.
Carvedilol is a potent noncardioselective beta-blocker, with weak vasodilating properties because of alpha 1 blockade. A reduction in both intrahepatic and portocollateral resistance contribute to enhanced effects on portal pressure. There are 10 published hemodynamic studies involving 168 patients investigating the role of carvedilol in portal hypertension. A reduction in the hepatic venous pressure gradient of up to 43% (range 10-43%) has been reported, particularly after chronic administration. However, tolerability at doses greater than 12.5 mg/day was comprised because of a fall in mean arterial pressure (MAP), particularly in ascitic patients. Carvedilol was more effective than propranolol in reducing hepatic venous pressure gradient in two of three studies, albeit with a greater decrease in MAP. One study showed deterioration of pre-existing ascites with carvedilol. The addition of nitrates to propranolol was less effective than carvedilol monotherapy in another study. A large multicentre, randomized controlled trial comparing carvedilol with variceal band ligation for the prevention of variceal bleeding has been published. Carvedilol resulted in fewer episodes of bleeding, although there was no difference in survival. Carvedilol was well tolerated. Carvedilol is a promising agent, and seems to be more effective than propranolol in hemodynamic studies. The efficacy in primary prevention of variceal bleeding suggests that carvedilol has a role in the management of clinically significant portal hypertension.


The critical role of health promotion for effective universal health coverage.
The Political Declaration from the United Nations High-Level Meeting on Universal Health Coverage: Moving Together to Build a Healthier World (2019) provided important reaffirmation of health as a precondition for sustainable development and equity, as well as of the role for primary care as a cornerstone of universal health coverage. Health promotion, prevention and sustainable healthcare go hand in glove. Health promotion can enable more effective use of health resources by reducing demand for expensive health services and reducing hospital admissions. Promoting mental and physical health, and addressing health literacy and the social determinants of health, enables governments and departments of health to (i) empower citizens and communities to take control of their health, and (ii) better support innovative and financially sustainable healthcare. Without the bedrock underpinning of effective health promotion, treasuries and health systems will struggle to meet the rising costs and burden of ill health.


NMDA receptor-dependent processes in the medial prefrontal cortex are important for acquisition and the early stage of consolidation during trace, but not delay eyeblink conditioning.
Permanent lesions in the medial prefrontal cortex (mPFC) affect acquisition of conditioned responses (CRs) during trace eyeblink conditioning and retention of remotely acquired CRs. To clarify further roles of the mPFC in this type of learning, we investigated the participation of the mPFC in mnemonic processes both during and after daily conditioning using local microinfusion of the GABA(A) receptor agonist muscimol or the NMDA receptor antagonist APV into the rat mPFC. Muscimol infusions into the mPFC before daily conditioning significantly retarded CR acquisition and reduced CR expression if applied after sufficient learning. APV infusion also impaired acquisition of CRs, but not expression of well-learned CRs. When infusions were made immediately after daily conditioning, acquisition of the CR was partially impaired in both the muscimol and APV infusion groups. In contrast, rats that received muscimol infusions 3 h after daily conditioning exhibited improvement in their CR performance comparable to that of the control group. Both the pre- and post-conditioning infusion of muscimol had no effect on acquisition in the delay paradigm. These results suggest that the mPFC participates in both acquisition of a CR and the early stage of consolidation of memory in trace, but not delay eyeblink conditioning by NMDA receptor-mediated operations.


Pain related behaviour during vincristine-induced neuropathy in rats.
The development of suitable animal models of neuropathic pain is essential to understand the pathophysiological mechanisms responsible for this condition. This study presents the alterations in nociception observed in rats suffering from a peripheral neuropathy induced by 10 daily repeated intravenous injections of vincristine at doses of 50 or 75 microg/kg (total dose 500 or 750 microg/ kg). The rats present both mechanical hyperalgesia, allodynia and a loss of sensitivity (thermal hypoalgesia). Conservation of good health, the fast appearance of symptoms which correspond well with human responses and the easy induction of nociceptive symptoms are favourable criteria for using this model at 50 microg/kg vincristine dose in the future.


Morpho-metabolic post-surgical patterns of non-infected prosthetic heart valves by [18F]FDG PET/CTA: "normality" is a possible diagnosis.
To define characteristic PET/CTA patterns of FDG uptake and anatomic changes following prosthetic heart valves (PVs) implantation over time, to help not to misdiagnose post-operative inflammation and avoid false-positive cases. Prospective evaluation of 37 post-operative patients without suspected infection that underwent serial cardiac PET/CTA examinations at 1, 6, and 12 months after surgery, in which metabolic features (FDG uptake distribution pattern and intensity) and anatomic changes were evaluated. Standardized uptake values (SUVs) were obtained and a new measure, the valve uptake index (VUI), (SUVmax-SUVmean)/SUVmax, was tested to homogenize SUV results.In total, 111 PET/CTA scans were performed in 37 patients (19 aortic and 18 mitral valves). FDG uptake was visually detectable in 79.3% of patients and showed a diffuse, homogeneous distribution pattern in 93%. Quantitative analysis yielded a mean maximum standardized uptake value (SUVmax) of 4.46 ± 1.50 and VUI of 0.35 ± 0.10. There were no significant differences in FDG distribution or uptake values between 1, 6, or 12 months. No abnormal anatomic changes or endocarditis lesions were detected in any patient during follow-up. FDG uptake, often seen in recently implanted PVs, shows a characteristic pattern of post-operative inflammation and, in the absence of associated anatomic lesions, could be considered a normal finding. These features remain stable for at least 1 year after surgery, so questioning the recommended 3-month safety period. A new measure, the VUI, can be useful for evaluating the FDG distribution pattern.


Aurora A is differentially expressed in gliomas, is associated with patient survival in glioblastoma and is a potential chemotherapeutic target in gliomas.
Aurora A is critical for mitosis and is overexpressed in several neoplasms. Its overexpression transforms cultured cells, and both its overexpression and knockdown cause genomic instability. In transgenic mice, Aurora A haploinsufficiency, not overexpression, leads to increased malignant tumor formation. Aurora A thus appears to have both tumor-promoting and tumor-suppressor functions. Here, we report that Aurora A protein, measured by quantitative protein gel blotting, is differentially expressed in major glioma types in lineage-specific patterns. Aurora A protein levels in WHO grade II oligodendrogliomas (n=16) and grade III anaplastic oligodendrogliomas (n=16) are generally low, similar to control epilepsy cerebral tissue (n=11). In contrast, pilocytic astrocytomas (n=6) and ependymomas (n=12) express high Aurora A levels. Among grade II to grade III astrocytomas (n=7, n=14, respectively) and grade IV glioblastomas (n=31), Aurora A protein increases with increasing tumor grade. We also found that Aurora A expression is induced by hypoxia in cultured glioblastoma cells and is overexpressed in hypoxic regions of glioblastoma tumors. Retrospective Kaplan-Meier analysis revealed that both lower Aurora A protein measured by quantitative protein gel blot (n=31) and Aurora A mRNA levels measured by real-time quantitative RT-PCR (n=58) are significantly associated with poorer patient survival in glioblastoma. Furthermore, we report that the selective Aurora A inhibitor MLN8237 is potently cytotoxic to glioblastoma cells, and that MLN8237 cytotoxicty is potentiated by ionizing radiation. MLN8237 also appeared to induce senescence and differentiation of glioblastoma cells. Thus, in addition to being significantly associated with survival in glioblastoma, Aurora A is a potential new drug target for the treatment of glioblastoma and possibly other glial neoplasms.


Is tailored adjuvant treatment for colon cancer possible?
High-risk stage II and stage III colon cancers are associated with significant recurrence rates after surgical resection. Adjuvant chemotherapy has demonstrated a significant disease-free and overall survival benefit for patients, even if not helpful for all. Currently, there are limited clinical and molecular markers that can predict response to chemotherapy and clinical outcome in advanced colon cancer. Herein, we review and discuss recent data about the most promising clinicopathologic and molecular factors described thus far and that are of interest in the management of patients with colon cancer.


Factors associated with stable high blood pressure in adolescents.
Our aim was to study the relative importance of the factors associated with stable high blood pressure (BP) in adolescents. Out of 17 634 children born in 1964 and screened in 1977, two samples, an upper group [the upper 5% of the distribution curves for both systolic (SBP) and diastolic (DBP) blood pressures] and a lower group (a 10% random sample from the remainder), were chosen for yearly follow-up (1680 boys and 1143 girls) from 1978 to 1981. A multiple logistic function was used to analyse the factors associated with stable high SBP and DBP (with at least three different measurements of BP at or above the 95th percentile). Different sets of variables were studied. Of the boys with stable high SBP, 55% were found in the upper quintile of risk distribution when Quetelet's index was taken into consideration, 63% when Quetelet's index + age + heart rate+sexual maturation, 67% when Quetelet's index + age + heart rate + serum uric acid and 74% when Quetelet's index + age + heart rate + cardiothoracic index + heart volume index were taken into consideration. Of the girls with a stable high SBP, 68% were found in the upper quintile of risk distribution when any of the following variables was added to Quetelet's index + age + heart rate: R and S waves, serum cholesterol or post-load blood sugar. Similar but smaller effects of associated factors on DBP were observed. Our study shows that Quetelet's index and heart rate are the most important factors associated with stable high BP in adolescents.


Misperceptions and inadequate pain management in cancer patients.
This article examines misperceptions and barriers to adequate pain relief in cancer patients. Healthcare professionals have gaps in their knowledge of opioid drugs as well as misconceptions concerning tolerance, physical dependence, and addiction that often lead to the underprescribing of these agents. The pervasiveness of the "say no to drugs" message in our society and the fear of addiction on the part of patients and their families creates yet another barrier to the legitimate use of opioids to treat cancer pain. Legal and regulatory documents filled with arbitrary and ill-defined labels meant to promote the legitimate use of these drugs and curtail their misuse may instead intimidate healthcare professionals and negatively influence prescribing habits. Increased educational efforts for pharmacists and other healthcare professionals as well as the development of clinical role models and state cancer pain initiatives are cited as means to break down these barriers in order to achieve adequate pain relief for all cancer patients.


Spatially divergent trends of nitrogen versus phosphorus limitation across European forests.
Nitrogen (N) and phosphorus (P) are essential nutrients that widely limit plant growth in global terrestrial ecosystems. Rising atmospheric CO2 concentration generally stimulates terrestrial net primary productivity and consequently may cause or aggravate N and P limitation due to a dilution effect, but the spatial variation of temporal trends in N versus P limitation and its key regulating factors is poorly understood. Using the leaf N:P ratio of 15 dominant tree species as an indicator, we analysed the spatial variation of plot-level shift towards N or P limitation across 163 European forest plots during 1995-2017. Phosphorus limitation increased from 25% to 33% of the studied plots between 1995-1997 and 2015-2017, while N limitation occurred in a negligible number of plots. A major proportion (56%) of the plots showed no significant trend in leaf N:P ratio, implying no shifts in N versus P limitation status. In the remaining plots, 38% of the plots showed a significant increase of leaf N:P ratio and only 6% of the plots showed a significant decrease of leaf N:P ratio. The spatial variation in the rate of decrease in leaf N:P ratio was associated with a significant decrease in leaf N concentration and mainly explained by the rate of decrease in N deposition. In contrast, the spatial variation in the rate of increase in leaf N:P ratio was associated with a significant decrease in leaf P concentration and mainly explained by forest category (broadleaf vs. conifer), mean annual temperature and soil C:N ratio. Our findings highlight a remarkable spatial divergence in temporal trends of nutrient limitation status across European forests over the past two decades, but overall, P is becoming more limiting versus N, especially in broadleaved forests.


Revalidating the Usefulness of a "Sector-Wise Regression" Approach to Predict Glaucomatous Visual Function Progression.
We previously developed a method to measure visual function progression using "Progression sectors," which were derived from a clustering analysis of pointwise rates of glaucomatous visual field (VF) progression; however, only short series of VFs, where pointwise linear regression (PLR) is typically not reliable, were analyzed. The purpose of the current study is to further investigate the usefulness of a sector-wise approach in longer series of VFs where the accuracy of PLR is inherently greater. We analyzed 16 VFs from each of 71 eyes in 45 patients. Sectorial averages of total deviation (TD) values were calculated at each "Progression sector," consisting of 23 sectors as well as at "Nouri-Mahdavi" (NM) sectors, consisting of 10 sectors. Linear regression then was applied to predict sectorial averages in the 11th to 16th VFs and absolute prediction errors were compared to PLR. Absolute prediction errors associated with linear regression of the "Progression sectors" were significantly smaller than those from PLR (P < 0.05) when predicting values in the 14th to 16th VFs. Conversely, prediction errors with NM sectors were significantly larger than those from PLR and "Progression sectors" (P < 0.01) when predicting values in the 11th to 16th VFs. It is useful to use "Progression sectors" to predict future VF progression in short (1-10 VFs) and long (>10 VFs) VF series, compared to PLR and NM sectors.


The Molecular Organization of Human cGMP Specific Phosphodiesterase 6 (PDE6): Structural Implications of Somatic Mutations in Cancer and Retinitis Pigmentosa.
In the cyclic guanosine monophosphate (cGMP) signaling pathway, phosphodiesterase 6 (PDE6) maintains a critical balance of the intracellular concentration of cGMP by catalyzing it to 5' guanosine monophosphate (5'-GMP). To gain insight into the mechanistic impacts of the PDE6 somatic mutations that are implicated in cancer and retinitis pigmentosa, we first defined the structure and organization of the human PDE6 heterodimer using computational comparative modelling. Each subunit of PDE6αβ possesses three domains connected through long α-helices. The heterodimer model indicates that the two chains are likely related by a pseudo two-fold axis. The N-terminal region of each subunit is comprised of two allosteric cGMP-binding domains (Gaf-A & Gaf-B), oriented in the same way and interacting with the catalytic domain present at the C-terminal in a way that would allow the allosteric cGMP-binding domains to influence catalytic activity. Subsequently, we applied an integrated knowledge-driven in silico mutation analysis approach to understand the structural and functional implications of experimentally identified mutations that cause various cancers and retinitis pigmentosa, as well as computational saturation mutagenesis of the dimer interface and cGMP-binding residues of both Gaf-A, and the catalytic domains. We studied the impact of mutations on the stability of PDE6αβ structure, subunit-interfaces and Gaf-cGMP interactions. Further, we discussed the changes in interatomic interactions of mutations that are destabilizing in Gaf-A (R93L, V141 M, F162 L), catalytic domain (D600N, F742 L, F776 L) and at the dimer interface (F426A, F248G, F424 N). This study establishes a possible link of change in PDE6αβ structural stability to the experimentally observed disease phenotypes.


A novel multi-unit tablet for treating circadian rhythm diseases.
This study aimed to develop and evaluate a novel multi-unit tablet that combined a pellet with a sustained-release coating and a tablet with a pulsatile coating for the treatment of circadian rhythm diseases. The model drug, isosorbide-5-mononitrate, was sprayed on microcrystalline cellulose (MCC)-based pellets and coated with Eudragit(®) NE30D, which served as a sustained-release layer. The coated pellets were compressed with cushion agents (a mixture of MCC PH-200/ MCC KG-802/PC-10 at a ratio of 40:40:20) at a ratio of 4:6 using a single-punch tablet machine. An isolation layer of OpadryII, swellable layer of HPMC E5, and rupturable layer of Surelease(®) were applied using a conventional pan-coating process. Central-composite design-response surface methodology was used to investigate the influence of these coatings on the square of the difference between release times over a 4 h time period. Drug release studies were carried out on formulated pellets and tablets to investigate the release behaviors, and scanning electron microscopy (SEM) was used to monitor the pellets and tablets and their cross-sectional morphology. The experimental results indicated that this system had a pulsatile dissolution profile that included a lag period of 4 h and a sustained-release time of 4 h. Compared to currently marketed preparations, this tablet may provide better treatment options for circadian rhythm diseases.


Stability of nifedipine in two oral suspensions stored at two temperatures.
To determine the stability of nifedipine in two oral suspensions stored at 4 degrees C and 25 degrees C in plastic prescription bottles over a 3-month period. Two suspensions were prepared: 1% methylcellulose:simple syrup NF (1:13) stored in ten 2 oz bottles; and Ora Plus:Ora Sweet (1:1) at a nifedipine concentration of 4 mg/mL also stored in ten 2 oz bottles. Five bottles of each formulation were stored at 4 degrees C, and the rest of the bottles were stored at 25 degrees C. Three samples were collected from each bottle at 0, 7, 14, 28, 42, 56, 70, and 91 days and analyzed by a stability-indicating high-performance liquid chromatography analytic method (n = 15). Laboratory. At 4 degrees C, the mean concentration of nifedipine was nearly 100% that of the original concentration in both the Ora Plus-Ora Sweet and the 1% methylcellulose-syrup formulations during the 3-month study period. At 25 degrees C, the mean +/- standard deviation concentration of nifedipine was 98.9% +/- 2.36% of the initial concentration in the Ora Plus-Ora Sweet and 97.4% +/- 2.48% in the 1% methylcellulose-syrup formulation over the 3-month period. No changes in pH or physical appearance were observed. Nifedipine can be prepared in two liquid dosage forms and stored for up to 3 months under refrigeration or at room temperature.


Role of muscarinic receptors in the activity of N-desmethylclozapine: reversal of hyperactivity in the phospholipase C knockout mouse.
Activity of the cholinergic muscarinic system is associated with modulation of locomotor activity, although the precise mechanism remains unclear. The phospholipase C-beta1 knockout mouse displays both M1 muscarinic receptor dysfunction and a hyperactive locomotor phenotype. This mouse serves as an ideal model for the analysis of muscarinic modulation of locomotor activity. The clozapine metabolite N-desmethylclozapine (NDMC) has shown some promise as an alternative or adjunct treatment for psychotic disorders. NDMC shows strong muscarinic acetylcholine receptor affinities, which may contribute to the clinical efficacy of clozapine and account for the correlation between NDMC/clozapine ratio and treatment response. Administration of NMDC reversed a striking hyperactive phenotype in the phospholipase C-beta1 knockout mouse, whereas no significant effects were observed in wild-type animals. This highlights the potential role of muscarinic activity in the behavioural response to NDMC. The M1 muscarinic antagonist pirenzepine, however, also reduced the hyperactive phenotype of these mice, emphasizing the importance of muscarinic function in the control of locomotor behaviour, but also calling into question the specific mechanism of action of NMDC at muscarinic receptors.