Docking flexible ligands in proteins with a solvent exposure- and distance-dependent dielectric function.
Physics-based force fields for ligand-protein docking usually determine electrostatic energy with distance-dependent dielectric (DDD) functions, which do not fully account for the dielectric permittivity variance between approximately 2 in the protein core and approximately 80 in bulk water. Here we propose an atom-atom solvent exposure- and distance-dependent dielectric (SEDDD) function, which accounts for both electrostatic and dehydration energy components. Docking was performed using the ZMM program, the AMBER force field, and precomputed libraries of ligand conformers. At the seeding stage, hundreds of thousands of positions and orientations of conformers from the libraries were sampled within the rigid protein. At the refinement stage, the ten lowest-energy structures from the seeding stage were Monte Carlo-minimized with the flexible ligand and flexible protein. A search was considered a success if the root mean square deviation (RMSD) of the ligand atoms in the apparent global minimum from the x-ray structure was <2 A. Calculations on an examining set of 60 ligand-protein complexes with different DDD functions and solvent-exclusion energy revealed outliers in most of which the ligand-binding site was located at the protein surface. Using a training set of 16 ligand-protein complexes, which did not overlap with the examining set, we parameterized the SEDDD function to minimize the RMSD of the apparent global minima from the x-ray structures. Recalculation of the examining set with the SEDDD function demonstrated a 20% increase in the success rate versus the best-performing DDD function.


A comparison of iron absorption in adults and infants consuming identical infant formulas.
Fe absorption was estimated in adults and infants from the erythrocyte incorporation of Fe isotopes added to infant formula. Fe absorption was measured in adults using radioisotopes, and in infants with a stable-isotope technique. In adults, the geometric mean Fe absorption from a ready-to-feed soya formula with its native phytic acid content was 2.4%. This increased to 6.0% (P < 0.05) after almost complete dephytinization. In infants, mean Fe absorption values were 3.9 and 8.7% respectively from the same products (P < 0.05). In adults, mean Fe absorption from a spray-dried soya formula containing 110 mg ascorbic acid/l was 4.1%, increasing to 5.3% (P < 0.05) when ascorbic acid was doubled to 220 mg/l. In infants, mean Fe absorption values were 5.7 and 9.5% (P < 0.05) from the same products. Mean Fe absorption from a milk-based formula was 6.5% in adults compared with 6.7% in infants. All meals in the adult and infant studies were fed using an identical meal size of 217 g. Increasing the meal size threefold in adults did not change fractional Fe absorption. Mean Fe absorption values for each meal were lower in adults than in infants but the relative inhibitory effect of phytic acid and the enhancing effect of ascorbic acid were similar. We conclude that Fe absorption studies in adults can be used to assess the influence of enhancers and inhibitors of Fe absorption in infant formulas fed to infants. Further studies, however, are required to extend these findings to weaning foods and complete meals.


Shear bond strength of a composite resin after delayed bonding to a polymerised dentine bonding system.
Polymerised adhesive may serve as a temporary sealant on prepared dentine surfaces and may be used as a precursor to the final bonding of a resin restoration. This procedure may, however, have an adverse effect on the strength of the final bond. This study was undertaken on 40 extracted human molar teeth to assess the effect of delayed bonding on the shear bond strength of a composite resin luted with a luting resin to abraded dentine treated with the Scotchbond Multi-Purpose (SMP) Bonding System (3M). No significant difference (p = 0.969) could be demonstrated between shear bond strengths achieved after immediate and delayed bonding to the polymerised Scotchbond Multi-Purpose adhesive.


Epidemiology of injury in elite and subelite female gymnasts: a comparison of retrospective and prospective findings.
An 18 month prospective injury survey was conducted on 64 Australian elite and subelite female gymnasts. The aims were to determine the rate of injury, anatomical location, and types of injury incurred by female competitive gymnasts, and to compare the findings with data collected retrospectively from the same sample of gymnasts. The gymnasts recorded (weekly) in an injury record booklet the number of hours trained and information on any injuries suffered over that week. The sample reported 349 injuries, a rate of 5.45 per person (6.29 for the elite and 4.95 for subelite gymnasts) over the 18 month survey. Injuries to the ankle and foot (31.2%) were the most commonly reported, followed by the lower back (14.9%). The most prevalent type of injury were sprains (29.7%), followed by strains (23.2%), and growth plate injuries (12.3%). The elite gymnasts reported that, for each injury, they missed fewer training sessions (p = 0.01), but modified more sessions (p = 0.0001) than their subelite counterparts. Further, the elite gymnasts spent 21.0% of the year training at less than full capacity because of injury. Although a significantly higher number of injuries were recorded in the prospective study (p = 0.0004), no differences were found between the distribution of injury by anatomical location or type between the two methods of data collection. The findings have important implications in terms of training procedures and periodic screening of gymnasts.


Advances in understanding and management of lymphoedema (cancer, primary).
Despite being neglected both clinically and in research in the past, primary lymphoedema, and lymphoedema secondary to cancer treatment have been the subject of an increasing number of studies in recent years. This review will focus on new developments relevant to clinical practice. The main themes covered by the review include the incidence and prevalence of lymphoedema, early detection and intervention in cancer-related lymphoedema, risk reduction for the development of cancer-related lymphoedema, surgical treatment, palliative care for lymphoedema in advanced cancer and developments in the genetics of primary lymphoedema. Evidence suggests that lymphoedema is more common than has been previously recognized. It continues to be a significant problem for people following cancer treatment. Developments in the early detection and treatment of cancer-related lymphoedema should reduce the future impact for patients. Advice on how to reduce the risk of developing lymphoedema may need to be modified in light of recent research. New treatments such as surgical procedures are still in their infancy but the results seem encouraging for selected patients.


Central retinal vein occlusion in a woman receiving bromocriptine.
We evaluated a unilateral retinal vascular abnormality in a 34-year-old woman receiving bromocriptine mesylate for the suppression of postpartum lactation. After using bromocriptine, 2.5 mg twice daily for 1 week, the patient complained of metamorphopsia in the left eye. A central retinal vein occlusion was found. We believe that central retinal vein occlusion in patients receiving bromocriptine, as demonstrated in our patient, is rare.


Inducible nitric oxide synthase inhibition improves intestinal microcirculatory oxygenation and CO2 balance during endotoxemia in pigs.
We examined whether selective inhibition of inducible nitric oxide synthase (iNOS) promotes intestinal microvascular oxygenation (microPO2) and CO2 off-load after endotoxic shock. Prospective, controlled experimental study in a university animal research laboratory. 13 domestic pigs. After baseline measurements shock was induced by 1 microg kg-1 h-1 endotoxin until mean arterial pressure fell below 60 mmHg. After 30 min in shock the animals were resuscitated with either fluid alone (control, n=6) or fluid and the iNOS inhibitor N-[3-(aminomethyl)benzyl]acetamidine hydrochloride (1400W, n=7). As final experimental intervention all animals received the nonselective NOS inhibitor L-NAME. Systemic and regional hemodynamic and oxygenation parameters were measured at baseline, during endotoxemia and shock, hourly for 3 h of 1400W therapy, and 30 min after the final L-NAME administration. microPO2 was assessed by the Pd-porphyrin phosphorescence technique, and the arterial to intestinal PCO2 gap was determined by air tonometry. Endotoxemia and shock resulted in a decrease in ileal mucosal and serosal microPO2 and a rise in PCO2 gap. The combination of 1400W and fluid resuscitation, but not fluid alone, normalized both the serosal microPO2 and the intestinal PCO2 gap. Administration of L-NAME decreased cardiac output and oxygen delivery and intestinal microPO2 and blood flow in both groups. Partial blockade of NO production by 1400W increased serosal microvascular oxygenation and decreased the intestinal CO2 gap. This findings are consistent with the idea that 1400W corrects pathological flow distribution and regional dysoxia within the intestinal wall following endotoxic shock.


The effect of crestal versus mucobuccal incisions on the success rate of implant osseointegration.
A retrospective analysis compared the success rate of osseointegration of Nobelpharma implants at stage 2 surgery. The implants had been placed using either a mucobuccal fold incision or a crestal incision. A total of 386 implants was placed in 92 patients; 265 implants were placed in 60 patients using a mucobuccal fold incision, with a success rate of 98.8%, and 121 implants were placed in 32 patients using a crestal incision, with a success rate of 98.3%. Implants can be placed using either a mucobuccal fold incision or a crestal incision to achieve predictable success.


Deoxyribonucleic acid restriction and modification systems in Salmonella: chromosomally located systems of different serotypes.
With the use of four different phages, Salmonella strains representing 85 different serotypes were examined to determine their restriction-modification phenotype. They fell into one of three groups on this basis: group 1, those which lacked the common LT system; group 2, those in which only the LT system could be recognized; and group 3. those which possessed the LT system and at least one other system shown with some serotypes to be closely linked to serB. The specificity of the serB-linked restriction-modification system was unique for each serotype, but different strains of the same serotype expressed the same specificity. Two of the systems were shown to behave in genetic crosses as functional alleles of the S. typhimurium SB system. It is possible that these serB-linked restriction-modification systems constitute a large multiallelic series of genes extending throughout the Salmonella genus and Escherichia coli. We suggest that the division of the Salmonella into the three restriction-modification groups may be significant in defining a "biological grouping" of the different serotypes within the genus which may ultimately be useful in describing the Salmonella species. From the genetic relatedness between the genes of some of the Salmonella restriction-modification systems with those of the E. coli systems, we deduce that the restriction endonuclases produced by the Salmonella serB-linked systems are of type 1. Determination of the nucleotide sequences of the recognition sites of the restriction endonucleases of selected Salmonella systems should further our understanding of specificity with these enzymes.


Fixed 30 mCi (1110 MBq) 131I-iodine therapy in autonomously functioning nodules: Single toxic nodule as a predictive factor of success.
The aim of this study is to evaluate the efficacy of a fixed 30 mCi (1110 MBq) 131I-iodine dose for the treatment of hyperthyroidism due to uninodular or multinodular toxic goiter and identify predictors of success. Fifty-nine patients diagnosed with nonautoimmune toxic goiter were treated with a fixed 30 mCi dose of 131I-iodine and were followed at a tertiary service between 2000 and 2016. The therapy was considered successful if the patient reached euthyroidism or hypothyroidism without needing an extra 131I-iodine dose or antithyroid drugs for at least 1 year after the radioiodine therapy (RIT). Patients with a single toxic nodule were younger at diagnosis (52 vs. 63 years; P = 0.007), presented a shorter disease duration until RIT (2 vs. 3.5 years; P = 0.007), smaller total thyroid volume (20 vs. 82 cm3; P = 0.044), and lower pre-RIT thyroid uptake (P = 0.043) than patients with multinodular goiter. No significant difference was seen with antithyroid drug use, thyroid-stimulating hormone and free thyroxine level, and follow-up after RIT. After RIT, 47 patients (79.66%) met the success criteria, and 12 (20.33%) remained hyperthyroid. Among the success group, 32 (68.08%) reached euthyroidism, while 31.92% developed hypothyroidism after 1 year. Patients with single toxic nodules who achieved success after RIT presented smaller nodules (2.8 vs. 5.75 cm; P = 0.043), while the pre-RIT thyroid uptake was higher among patients with multinodular toxic goiter who achieved success after RIT (5.5% vs. 1.5%; P = 0.007). A higher success rate was observed among patients with a single toxic nodule than those with a toxic multinodular goiter (92.3% vs. 55%; P = 0.001), and a single toxic nodule presentation was found to be an independent predictor of success (P = 0.009). The fixed 30 mCi 131I-iodine dose was particularly effective in the group of patients with single autonomously functioning nodule rather than the group with multiple nodules. A single toxic nodule was an independent predictor of treatment success.


Affective disorders and mortality. A general population study.
A 16-year prospective study of a general population sample indicates that those who had reported a depression and/or anxiety disorder at baseline experienced 1.5 times the number of deaths expected on the basis of rates for a large reference population. As part of the Stirling County Study (Canada), the information was gathered from 1003 adults through structured interviews and was analyzed by means of a diagnostic computer program. The risk for mortality was assessed using external and internal standards, controlling for the effects of age and sex as well as for the presence of self-reported physical disorders at baseline. Increased risk was found to be significantly associated with affective but not physical disorders and with depression but not generalized anxiety. When this evidence about mortality was combined with information about subsequent psychiatric morbidity among survivors, 82% of those who were depressed at baseline had a poor outcome.


Propranolol and periocular capillary hemangiomas: assessment of refractive effect.
To assess the effects of systemic propranolol on refractive error in infants with periocular capillary hemangiomas. A single-center study of consecutive patients with capillary hemangiomas treated with systemic propranolol. Refractive data were analyzed using Long's matrix formalism and the methods of Harris and Kaye. Seventeen patients were included. At 6 months postoperatively, hemangioma size reduced from 3,214 to 1,806 mm(3) (standard deviation: 4,122 to 2,441). Mean refractive error in the affected eye significantly reduced: -1.25/0.38 × 36 (95% confidence intervals: -5.08/1.20 × 90 to 1.64/1.43 × 180, P = .048) with a smaller change (P = .06) in the unaffected eye of -1.01/+0.31 × 3.16 (95% confidence intervals: -4.02/+1.12 × 180 to +1.49/+0.51 × 90). Propranolol produced a clinically significant reduction in the infants' refractive error and anisometropia. The reduction in the total refractive error and anisometropia has not been evident in previous analyses, which have concentrated on the change in the "cylinder" as the principal outcome measure.


Effect of pelvic floor muscle exercise on pelvic floor muscle activity and voiding functions during pregnancy and the postpartum period.
The aim of this study was to investigate the effects of pelvic floor muscle exercise during pregnancy and the postpartum period on pelvic floor muscle activity and voiding functions. Pregnant women (n = 60) were randomly assigned into two groups (Training [n = 30] and Control [n = 30]) using a computer-based system. Pelvic floor muscle strength was measured using a perineometry device. Urinary symptoms were measured using the Urinary Distress Inventory (UDI-6), Incontinence Impact Questionnaire (IIQ-7), and the Overactive Bladder Questionnaire (OAB-q). Voiding functions were measured using uroflowmetry and 3-day voiding diaries. Measurements were obtained at week 28, weeks 36-38 of pregnancy, and postpartum weeks 6-8. Pelvic floor muscle strength significantly decreased during the pregnancy (P < 0.001). However, pelvic floor muscle strength improvement was significantly higher in the Training group compared to the Control group (P < 0.001). The UDI-6, IIQ-7, and OAB-q scores did not significantly change during weeks 36-38 of pregnancy in the Training group (P > 0.05). However, UDI-6, coping, concern, and total scores of OAB-q were significantly decreased during weeks 36-38 of pregnancy in the Control group (P < 0.05). The UDI-6 and OAB-q scores were significantly improved during postpartum weeks 6-8 (P < 0.05). Voiding functions were negatively affected in both groups, decreasing during weeks 36-38 of pregnancy and improving during the postpartum period. Pregnancy and delivery affect pelvic floor muscle strength, urinary symptoms, quality of life, and voiding functions. Pelvic floor muscle exercises applied during pregnancy and the postpartum period increase pelvic floor muscle strength and prevent deterioration of urinary symptoms and quality of life in pregnancy.


Carbon Monoxide Preserves Circadian Rhythm to Reduce the Severity of Subarachnoid Hemorrhage in Mice.
Subarachnoid hemorrhage (SAH) is associated with a temporal pattern of stroke incidence. We hypothesized that natural oscillations in gene expression controlling circadian rhythm affect the severity of neuronal injury. We moreover predict that heme oxygenase-1 (HO-1/Hmox1) and its product carbon monoxide (CO) contribute to the restoration of rhythm and neuroprotection. Murine SAH model was used where blood was injected at various time points of the circadian cycle. Readouts included circadian clock gene expression, locomotor activity, vasospasm, neuroinflammatory markers, and apoptosis. In addition, cerebrospinal fluid and peripheral blood leukocytes from SAH patients and controls were analyzed for clock gene expression. Significant elevations in the clock genes Per-1, Per-2, and NPAS-2 were observed in the hippocampus, cortex, and suprachiasmatic nucleus in mice subjected to SAH at zeitgeber time (ZT) 12 when compared with ZT2. Clock gene expression amplitude correlated with basal expression of HO-1, which was also significantly greater at ZT12. SAH animals showed a significant reduction in cerebral vasospasm, neuronal apoptosis, and microglial activation at ZT12 compared with ZT2. In animals with myeloid-specific HO-1 deletion (Lyz-Cre-Hmox1fl/fl ), Per-1, Per-2, and NPAS-2 expression was reduced in the suprachiasmatic nucleus, which correlated with increased injury. Treatment with low-dose CO rescued Lyz-Cre-Hmox1fl/fl mice, restored Per-1, Per-2, and NPAS-2 expression, and reduced neuronal apoptosis. Clock gene expression regulates, in part, the severity of SAH and requires myeloid HO-1 activity to clear the erythrocyte burden and inhibit neuronal apoptosis. Exposure to CO rescues the loss of HO-1 and thus merits further investigation in patients with SAH.


The relationship between hamstring muscle extensibility and spinal postures varies with the degree of knee extension.
The aim was to determine the relationship between hamstring muscle extensibility and sagittal spinal curvatures and pelvic tilt in cyclists while adopting several postures. A total of 75 male cyclists were recruited for this study (34.79 ± 9.46 years). Thoracic and lumbar spine and pelvic tilt were randomly measured using a Spinal Mouse. Hamstring muscle extensibility was determined in both legs by a passive knee extension test. Low relationships were found between hamstring muscle extensibility and spinal parameters (thoracic and lumbar curvature, and pelvic tilt) in standing, slumped sitting, and on the bicycle (r = .19; P > .05). Significant but low relationships were found in maximal trunk flexion with knees flexed (r = .29; P < .05). In addition, in the sit-and-reach test, low and statistically significant relationships were found between hamstring muscle extensibility for thoracic spine (r = -.23; P = .01) and (r = .37; P = .001) for pelvic tilt. In conclusion, hamstring muscle extensibility has a significant relationship in maximal trunk flexion postures with knees flexed and extended, but there are no relationships while standing or on the bicycle postures.


Evolution of the sweet taste receptor gene Tas1r2 in bats.
Taste perception is an important component of an animal's fitness. The identification of vertebrate taste receptor genes in the last decade has enabled molecular genetic studies of the evolution of taste perception in the context of the ecology and dietary preferences of organisms. Although such analyses have been conducted in a number of species for bitter taste receptors, a similar analysis of sweet taste receptors is lacking. Here, we survey the sole sweet taste-specific receptor gene Tas1r2 in 42 bat species that represent all major lineages of the order Chiroptera, one of the most diverse groups of mammals in terms of diet. We found that Tas1r2 is under strong purifying selection in the majority of the bats studied, with no significant difference in the strength of the selection between insect eaters and fruit eaters. However, Tas1r2 is a pseudogene in all three vampire bat species and the functional relaxation likely started in their common ancestor, probably due to the exclusive feeding of vampire bats on blood and their reliance on infrared sensors rather than taste perception to locate blood sources. Our survey of available genome sequences, together with previous reports, revealed additional losses of Tas1r2 in horse, cat, chicken, zebra finch, and western clawed frog, indicating that sweet perception is not as conserved as previously thought. Nonetheless, we found no common dietary pattern among the Tas1r2-lacking vertebrates, suggesting different causes for the losses of Tas1r2 in different species. The complexity of the ecological factors that impact the evolution of Tas1r2 calls for a better understanding of the physiological roles of sweet perception in different species.


Relationship between lunar cycle and haemorrhagic complication rate in surgery.
The aim of this study was to evaluate a possible relationship between lunar cycles and haemorrhagic complication rate in surgery. The possible relationship between moon phases and surgical outcome was tested by evaluating the haemorrhagic complication rate for 18,760 patients who underwent surgery between January 2001 and December 2008 at the National Institute for Cancer Research in Genoa. A total of 103 lunar phases were considered using Chi-square (χ2) test analysis, and patients were allocated a surgery date. One hundred and sixty-seven haemorrhagic complications were observed. Three hundred and nine new moon phase days were analysed and 12 incidences of complications detected, with a 3.9% complication rate per day. In the waxing moon phase, 1184.5 d were analysed with 68 incidences of complications at a daily rate of 5.7%. In the full moon phase there was a 4.9% complication rate per day (15 incidences in 309 d), whereas in the waning moon phase, the 6% percentage rate per day resulted from 72 incidences in 1184.5 d. No statistically significant correlations were found between moon cycles and postoperative haemorrhagic complications (p = .50).


The Jumonji-domain histone demethylase inhibitor JIB-04 deregulates oncogenic programs and increases DNA damage in Ewing Sarcoma, resulting in impaired cell proliferation and survival, and reduced tumor growth.
Ewing Sarcoma is an aggressive malignant neoplasm affecting children and young adults. Ewing Sarcoma is driven by transcription factor fusion oncoproteins, most commonly EWS/Fli1. While some patients can be cured with high-dose, multi-agent, chemotherapy, those that cannot currently have few options. Targeting of the driver oncofusion remains a logical therapeutic approach, but has proven difficult. Recent work has pointed to epigenetic mechanisms as key players, and potential new therapeutic targets, in Ewing Sarcoma. In this study we examined the activity of the pan-JHDM pharmacologic inhibitor JIB-04 in this disease. We show that JIB-04 potently inhibits the growth and viability of Ewing Sarcoma cells, and also impairs tumor xenograft growth. Effects on histone methylation at growth-inhibitory doses vary among cell lines, with most cell lines exhibiting increased total H3K27me3 levels, and some increased H3K4me3 and H3K9me3. JIB-04 treatment widely alters expression of oncogenic and tumor suppressive pathways, including downregulation of known oncogenic members of the Homeobox B and D clusters. JIB-04 also disrupts the EWS/Fli1 expression signature, including downregulation of pro-proliferative pathways normally under positive oncofusion control. Interestingly, these changes are accompanied by increased levels of the EWS/Fli1 oncofusion, suggesting that the drug could be uncoupling EWS/Fli1 from its oncogenic program. All Ewing Sarcoma cell lines examined also manifest increased DNA damage upon JIB-04 treatment. Together, the findings suggest that JIB-04 acts via multiple mechanisms to compromise Ewing Sarcoma cell growth and viability.


Submuscular Versus Subcutaneous Ulnar Nerve Transposition: A Cadaveric Model Evaluating Their Role in Primary Ulnar Nerve Repair at the Elbow.
To investigate the length gained from subcutaneous and submuscular transposition of the ulnar nerve at the elbow. Specifically, the study aimed to define an expected nerve gap able to be overcome, and to determine if a difference between transposition techniques exists. Eleven cadaveric specimens from the scapula to fingertip were procured. In situ decompression and mobilization of the ulnar nerve at the elbow followed by simulated laceration of the nerve was performed. Nerves were marked 5 mm from the laceration site to facilitate overlap measurement and to simulate nerve end preparation to viable fascicles before primary coaptation. Nerve ends were attached to spring gauges set at 100 g of tension (strain ≤ 10%). Measurements of nerve overlap were obtained in varying degrees of wrist (0°, 30°, 60°) and elbow (0°, 15°, 30°, 45°, 60°, 90°) flexion. Measurements were performed after in situ decompression and mobilization, and then repeated after both subcutaneous and submuscular transposition. Ulnar nerve transposition was found to increase nerve overlap at an elbow flexion of 30° or greater. No difference was seen between subcutaneous and submuscular transpositions at all wrist and elbow positions. In situ decompression and mobilization alone provided an average of 3.5 cm of length gain with the elbow extended. Transposition in conjunction with clinically feasible wrist and elbow flexion (30° and 60°, respectively) provided 5.2 cm of length gain. Controlling for mobilization, a statistically significant increase in overlap of approximately 2 cm was gained from transposition. Although mobilization combined with wrist and elbow flexion may afford substantial gap reduction and should be used initially when approaching proximal ulnar nerve lacerations, transposition should be considered when faced with a large nerve gap greater than 3 cm at the elbow. No difference was seen between submuscular and subcutaneous transposition techniques. This study defines the extent an ulnar nerve gap at the elbow can be overcome by in situ mobilization, joint positioning, and transposition. It additionally compares the efficacy of submuscular and subcutaneous transposition techniques in closing this gap.


Tropical splenomegaly syndrome in a nontropical setting.
Tropical splenomegaly syndrome, a rare complication of recurrent malarial infection thought to occur only in endemic areas, was diagnosed in a 9-year resident of the United States. The patient had splenomegaly, anemia, a history of recurrent fever since childhood, cryoglobulinemia, increased serum IgM, and elevated specific immunofluorescent antibody titers to Plasmodium falciparum. After antimalarial treatment and splenectomy, she became asymptomatic and the IgM levels and specific antibody titers returned to normal. Because of increased travel to and from endemic malarial areas, this syndrome should be considered in the differential diagnosis of chronic splenomegaly.


Perinatal and maternal outcomes in women with sickle or hemoglobin C trait.
Recent studies have reported increased fetal loss and preeclampsia in women with sickle cell trait (hemoglobin [Hb] AS). There is a paucity of studies of outcomes in carriers of hemoglobin C. We examined the prevalence of hemoglobin C and S carrier status (Hb AC and Hb AS, respectively) and their effect on pregnancy outcomes. This was a retrospective cohort study using data prospectively collected from 1991 to 2006. Perinatal and maternal outcomes for African-American women with Hb AS and Hb AC were compared with those with normal hemoglobin (Hb AA). Multivariable regression was performed by applying generalized estimating equations to account for correlation between births from the same woman. Among 22,096 eligible African-American women (36,897 pregnancies) with routine antenatal hemoglobin electrophoresis, 88.5% had a normal (Hb AA) pattern. Hemoglobin AS was identified in 8.2% and Hb AC in 2.4% of women. Hemoglobin SS and Hb SC each accounted for less than 0.2% and Hb CC for 0.01%. Prevalence and relative risks for adverse outcomes in 3,019 AS pregnancies (3,062 births) and 875 AC (886 births), compared with 32,724 AA pregnancies (33,213 births), were not increased. Adjusted relative risks (95% confidence intervals) for perinatal mortality and preeclampsia were 0.7 (0.5-1.0) and 1.0 (0.8-1.2), respectively, for AS and 0.7 (0.3-1.4) and 1.0 (0.6-1.3), respectively, for AC. Risks of stillbirths and pregnancy-associated hypertension were also not increased. Contrary to other recent reports, perinatal mortality and preeclampsia are not increased in carriers of sickle cell trait or hemoglobin C. II.


Involvement of the membrane lipid bilayer in sorting prohormone convertase 2 into the regulated secretory pathway.
Prohormone convertase 2 (PC2) is a neuroendocrine-specific protease involved in the intracellular maturation of prohormones and proneuropeptides. PC2 is synthesised as a proprotein (proPC2) that undergoes proteolysis, aggregation and membrane association during its transit through the regulated secretory pathway. We have previously shown that the pro region of proPC2 plays a key role in its aggregation and membrane association. To investigate this further, we determined the binding properties of a peptide containing amino acids 45-84 of proPC2 (proPC2(45-84)) to trans-Golgi network/granule-enriched membranes from the AtT20 cell line. Removal of peripheral membrane proteins or hydrolysis of integral membrane proteins did not affect the binding properties of proPC2(45-84). Rather, proPC2(45-84) was shown to bind to protein-free liposomes in a pH- and Ca(2+)-dependent manner. To identify the component of the lipid bilayer involved in this membrane association, we used chromaffin-granule membranes and studied the binding properties of the endogenous PC2. Treatment of the membranes with saponin, a cholesterol-depleting detergent, failed to extract PC2 from the membranes, whereas chromogranin A (CgA) was removed. Treatment of the membranes with Triton X-100 yielded a low-density detergent-insoluble fraction enriched in PC2, but not CgA. The detergent-insoluble fraction also contained glycoprotein III, known to be part of the lipid rafts (membrane microdomains rich in sphingolipids). Finally, sphingolipid depletion of AtT20 cells resulted in the mis-sorting of PC2, suggestive of a link between the association of PC2 with lipid rafts in the membrane and its sorting into the regulated secretory pathway.


[Lumbar epidural anesthesia, 1931-1936: a second debut].
Fidel Pagés Miravé (1886-1923) was a well-known Spanish military surgeon and among the first in the world to pioneer lumbar epidural anesthesia in 1920. Between 1931 and 1936, surgeons of various nationalities spread the use of lumbar epidural anesthesia worldwide. This second debut of metameric anesthesia has given us amusing anecdotes and stories of singular old- and new-world personalities who left a wealth of literature that deserves to be reviewed and valued. This review will use their original words and idiomatic expressions which, though archaic, have beauty and significance. Special emphasis is put on dates in comparisons. Because well-known names have been associated with lumbar epidural anesthesia, this review closes with the biographies of illustrious European and Latin American surgeons who played a major role in spreading the technique in the United States of America and other parts of the world.


Pharmacodynamic and pharmacokinetics of BW 825C: a new antihistamine.
The new H1-receptor antagonist BW 825C and triprolidine (2.5 and 5 mg) were administered to 12 healthy male volunteers in a double blind placebo controlled, balanced, crossover design. Histamine antagonism was measured by assessment of flare and weal areas after intradermal injection of histamine. The 2 compounds were approximately equipotent in blocking the flare and weal response to intradermal histamine and had a similar duration of action. Triprolidine impaired performance of vigilance and reaction time (p less than 0.05) compared with placebo while BW 825C did not. Drowsiness measured using visual analogue scales followed both triprolidine treatments, but not BW 825C. BW 825C had a plasma half-life (t1/2) of 1.7 +/- 0.2 h and triprolidine of 4.6 +/- 4.3 h. The peak plasma level of BW 825C was approximately 6 times that of triprolidine. It was concluded that BW 825C might be a clinically active H1-antagonist with reduced sedative side-effects.


A Single Tube Overlap Extension PCR Method for Splicing of Multiple DNA Fragments.
Despite the ease of conventional splicing by overlap-extension (SOEing) PCR technique in theory, when splicing more than two fragments, and especially if one of the complementary sequences is A-T rich, the attachment of the fragments would be challenging. A new rapid and highly efficient SOEing PCR assay was developed for simultaneous splicing of multiple DNA fragments and induction of site-directed mutagenesis in a single tube. The method was adapted for splicing human beta-globin UTRs to OCT4, SOX2, KLF4, C-MYC, LIN28A, and destabilized GFP for the construction of chimeric DNA fragments for in vitro transcription. In addition, the native Kozak sequence of beta-globin (K1) was replaced by the strongest Kozak sequence (K2) using site-directed mutagenesis to enhance the expression of target genes. ChimericGFPd2/K1, GFPd2/K2, OCT4, and KLF4 were created by the optimized conventional SOEing PCR. The single tube method was able to create the chimeric SOX2, C-MYC, and LIN28A in high quality and quantity in comparison with the conventional SOEing PCR. Moreover, using single tube SOEing PCR, the reaction time and materials that are required in the conventional SOEing PCR were significantly reduced. Fluorescent microscopy and flow cytometry examinations indicated highly efficient translation of K2 sequence in comparison with the K1sequence. Single tube SOEing PCR is a valuable method to construct more multiple fragments with high yield. The method can successfully be applied for construction of various kinds of complex chimeric genes.


Group III nitride nanomaterials for biosensing.
Biosensing has found wide applications in biological and medical research, and in clinical diagnosis, environmental monitoring and other analytical tasks. Recognized as novel and outstanding transducing materials because of their superior and unique physical/chemical properties, group III nitride (III-nitride) nanomaterials have been introduced into biosensor development with remarkable advancements achieved in the past few decades. This paper presents the first comprehensive review on biosensor development with III-nitride nanomaterials. The review starts with the introduction of the material properties and biocompatibility of III-nitrides that are useful for biosensing. The focus is then placed on surface treatments of III-nitrides, which lay the foundation for biosensing, and on biosensing mechanisms where the exceptional properties of III-nitride nanomaterials lead to superior biosensing performance. From a practical point of view, techniques for biosensor fabrication are then summarized. Finally, existing biosensing applications and future directions are discussed.


Neuropsychological correlates of behavioral symptoms in Alzheimer's disease, frontal variant of frontotemporal, subcortical vascular, and lewy body dementias: a comparative study.
The aim of this study was to investigate the neuropsychological correlates of behavioral and psychological symptoms (BPSD) in patients affected by various forms of dementia, namely Alzheimer's disease (AD), frontal-variant frontotemporal dementia (fvFTD), Lewy body dementia (LBD), and subcortical ischemic vascular dementia (SIVD). 21 fvFTD, 21 LBD, 22 AD, and 22 SIVD patients matched for dementia severity received a battery of neuropsychological tests and the Neuropsychiatry Inventory (NPI). The possible association between performance on neuropsychological tests and severity of BPSD was assessed by correlational analysis and multivariate regression. BPSD were present in 99% of patients. Most behavioral symptoms were not related to a particular dementia group or to a specific cognitive deficit. Euphoria and disinhibition were predicted by fvFTD diagnosis. Hallucinations correlated with the severity of visuospatial deficits in the whole sample of patients and were predicted by LBD diagnosis. Apathy, which was found in all dementia groups, correlated with executive functions and was predicted by both reduced set-shifting aptitude and fvFTD diagnosis. The results confirm the high prevalence of BPSD in the mild to moderate stages of dementia and show that most BPSD are equally distributed across dementia groups. Most of the cognitive and behavioral symptoms are independent dimensions of the dementia syndromes. Nevertheless, hallucinations in LBD and euphoria and disinhibition in fvFTD are related to the structural brain alterations that are responsible for cognitive decline in these dementia groups. Finally, apathy arises from damage in the frontal cortical areas that are also involved in executive functions.


Walking barefoot vs. with minimalist footwear - influence on gait in younger and older adults.
In recent years, minimalist footwear has been increasingly promoted for its use in sportive and recreational activities. These shoes are considered to function naturally like barefoot walking while providing a protective surface. Despite a growing popularity of these shoes in the older population, little is known about the influence of minimalist footwear on gait patterns. This study investigated whether overground walking with minimalist shoes is comparable to barefoot walking regarding gait stability and variability parameters. In a randomized within-subject study design, 31 healthy younger (29 ± 4 years) and 33 healthy community-dwelling older adults (71 ± 4 years) volunteered. Participants walked on flat ground, once barefoot and once with minimalist shoes. Gait variability of minimum toe clearance (MTC), stride length, stride time, and local dynamic gait stability were analysed. The results for both age groups showed significant condition effects (minimalist shoes vs. barefoot walking) for the outcomes of local dynamic stability (p = .013), MTC variability (p = .018), and stride length variability (p < .001) indicating increased local dynamic stability and decreased gait variability during the minimalist shoe condition. Group effects (young vs. older adults) were detected in all gait outcomes. Walking with minimalist shoes appeared to be associated with better gait performance than walking barefoot in both age groups. Thus, walking with minimalist shoes is not similar to barefoot walking. With respect to reducing the risk of falling, we suggest that minimalist shoes could be an alternative to barefoot walking or a transition option between shoes to barefoot for older adults.


Capitalizing on Cancer Specific Replication: Oncolytic Viruses as a Versatile Platform for the Enhancement of Cancer Immunotherapy Strategies.
The past decade has seen considerable excitement in the use of biological therapies in treating neoplastic disease. In particular, cancer immunotherapy and oncolytic virotherapy have emerged as two frontrunners in this regard with the first FDA approvals for agents in both categories being obtained in the last 5 years. It is becoming increasingly apparent that these two approaches are not mutually exclusive and that much of the therapeutic benefit obtained from the use of oncolytic viruses (OVs) is in fact the result of their immunotherapeutic function. Indeed, OVs have been shown to recruit and activate an antitumor immune response and much of the current work in this field centers around increasing this activity through strategies such as engineering genes for immunomodulators into OV backbones. Because of their broad immunostimulatory functions, OVs can also be rationally combined with a variety of other immunotherapeutic approaches including cancer vaccination strategies, adoptive cell transfer and checkpoint blockade. Therefore, while they are important therapeutics in their own right, the true power of OVs may lie in their ability to enhance the effectiveness of a wide range of immunotherapies.


Management of Adult Choledochal Cyst Coexisting with Gallbladder Carcinoma: A Case Report and Review of Literature.
Choledochal cyst is a relatively rare condition. Even rarer is a choledochal cyst in association with a gallbladder carcinoma. This study reports a rare case of gallbladder carcinoma coexisting with a choledochal cyst in a Nigerian patient. Clinical records of the patient including preoperative evaluation, intraoperative findings, and postoperative care were reviewed. A 38-year-old woman presented with the recurrent right upper abdominal pain of 3 years duration associated with progressive weight loss, anorexia, recurrent vomiting, as well as, low-grade fever with chills and rigors. Physical examination revealed an anicteric woman with tenderness in the right hypochondrium and a positive Murphy's sign. A combination of abdominal ultrasound and computed tomography scan suggested a Type IV choledochal cyst and a distended gallbladder with thickened walls containing a heterogeneous hyperdense mass. Preoperative serum alkaline phosphatase was elevated while endoscopic retrograde cholangiopancreatography was inconclusive. At laparotomy, extrahepatic biliary dilatation and enlarged, the nodular gallbladder was found with a diffusely fibrotic pancreas. Intraoperative cholangiogram confirmed Type IV choledochal cyst. Excision of the common bile duct and radical cholecystectomy was performed, and a Roux-en-Y hepaticojejunostomy. Histopathology confirmed the diagnosis of gallbladder adenocarcinoma. She had adjuvant chemotherapy and is presently on follow-up. No evidence of recurrence after 5 years of follow-up. A high index of suspicion is required to detect a combination of these two rare entities. When detected, both conditions should be surgically addressed at the same sitting, and when combined with adjuvant chemotherapy, may increase the chances of achieving a cure.


Unilateral neonatal testicular torsion.
Testicular torsion in the neonatal period is an unusual finding. Because of the high morbidity associated with this condition, early recognition and appropriate management are imperative. Testicular torsion may be unilateral or bilateral and may occur prenatally or in the early postnatal period. Often, the nurse is the first health care provider to examine a newborn after birth and must be aware of the signs of this condition to minimize the risk of testicular loss.


Effectiveness of non-adjuvanted pandemic influenza A vaccines for preventing pandemic influenza acute respiratory illness visits in 4 U.S. communities.
We estimated the effectiveness of four monovalent pandemic influenza A (H1N1) vaccines (three unadjuvanted inactivated, one live attenuated) available in the U.S. during the pandemic. Patients with acute respiratory illness presenting to inpatient and outpatient facilities affiliated with four collaborating institutions were prospectively recruited, consented, and tested for influenza. Analyses were restricted to October 2009 through April 2010, when pandemic vaccine was available. Patients testing positive for pandemic influenza by real-time RT-PCR were cases; those testing negative were controls. Vaccine effectiveness was estimated in logistic regression models adjusted for study community, patient age, timing of illness, insurance status, enrollment site, and presence of high-risk medical conditions. Pandemic virus was detected in 1,011 (15%) of 6,757 enrolled patients. Fifteen (1%) of 1,011 influenza positive cases and 1,042 (18%) of 5,746 test-negative controls had record-verified pandemic vaccination >14 days prior to illness onset. Adjusted effectiveness (95% confidence interval) for pandemic vaccines combined was 56% (23%, 75%). Adjusted effectiveness for inactivated vaccines alone (79% of total) was 62% (25%, 81%) overall and 32% (-92%, 76%), 89% (15%, 99%), and -6% (-231%, 66%) in those aged 0.5 to 9, 10 to 49, and 50+ years, respectively. Effectiveness for the live attenuated vaccine in those aged 2 to 49 years was only demonstrated if vaccination >7 rather than >14 days prior to illness onset was considered (61%∶ 12%, 82%). Inactivated non-adjuvanted pandemic vaccines offered significant protection against confirmed pandemic influenza-associated medical care visits in young adults.


[Sodium and potassium ion transport in the skin of Rana temporaria frogs under anaerobic conditions].
Saturation of incubation media with argon completely inhibits the unidirectional transport of Na+ across the frog skin while the ion composition of the skin cells does not change at all. The maintenance of the intracellular concentrations of Na+ and K+ cannot be explained by an assumption that at anaerobic conditions the cell membrane permeability to monovalent cations is markedly decreased. For instance, an addition of ouabain to the serosa bathing medium saturated with argon results in increasing the intracellular concentration of Na+ and loss of cell K+ indicating that both Na+ and K+ can freely diffuse across the cell membrane down to their electrochemical gradients. The results obtained suggest that the Na+/K+ pumps involved in the mechanism of unidirectional Na+ transport are critically dependent on the respiration energy while the pumps responsible for the cell ion homeostasis can operate at the expense of glycolysis. Compartmentalization of the energy sources and their coupling to consumers may explain independent functioning of the two mechanisms of Na+ transport developed in the course of cell evolution.


Support for people with dementia experiencing severe responsive behaviours: Unpacking the disconnect between policy and practice.
The Severe Behaviour Response Team (SBRT) program, which was established in 2015 to support aged care residents with dementia experiencing very severe and extreme responsive behaviours received far fewer referrals than projected during its first year. This article describes the outcomes of a brief survey to identify potential barriers to referrals and identify opportunities to improve the uptake of the service. A pragmatic, quasi-experimental study was conducted involving clinical leads working in a representative sample of care homes that had not used the SBRT. The study was part of the formative evaluation activities of an ongoing program evaluation. Of the 53 clinical leads that participated in the survey, one-third had not heard of the SBRT prior to being contacted. The remaining two-thirds (n = 36) had not used the service due to the availability of existing resources and concerns regarding responsiveness of, and access to, the new service. Three themes emerged from the study relating to awareness of the service, responsiveness and the interface between local aged care and health services. Referrals increased following interventions to address the first two themes; however, they continue to remain well below the number projected. This indicates a fundamental disconnection between the policy design process and the day-to-day experience of residential aged care. The study highlights the importance of aged care clinical leads being engaged in dementia policy and program development processes to support improved targeting of resources.


Cell trafficking, mediator release, and articular metabolism in acute inflammation of innervated or denervated isolated equine joints.
To describe the acute cellular response, inflammatory mediator release, and effect on chondrocyte metabolism of interleukin 1 beta (IL-1 beta) in isolated innervated or denervated equine metacarpophalangeal joints. One metacarpophalangeal joint of 24 adult horses. The metacarpophalangeal joint was isolated for 6 hours in a pump-perfused, auto-oxygenated, innervated or denervated metacarpophalangeal joint preparation. Isolated joints were assigned to 4 groups: control, control-denervated, inflamed, and inflamed-denervated, and inflammation was induced by intra-articular injection of IL-1 beta. Synovial fluid was collected for cytologic examination and determination of IL (IL)-1 beta, (IL-6), prostaglandin E2 (PGE2), and substance P (SP) values. Synovial membrane was immunostained with SP and nerve-specific enolase (NSE) antibodies. Cartilage was collected for determination of proteoglycan (PG) synthesis and degradation. IL-1 beta induced significant neutrophilic leukocytosis in synovial and synovial membrane. IL-1 beta concentration and returned to baseline by 5.5 hours, but IL-6 concentration significantly increased throughout the study. Total SP content was significantly higher in inflamed joints. There was a significant increase in 24- and 48-hour PG degradation in inflamed innervated joints. Cellular response to IL-1 beta was rapid and sustained; joint clearance of IL-1 beta was rapid, and endogenous production of IL-1 beta did not follow. The IL-6 and PGE2 concentrations significantly increased, and SP content was increased in association with inflammation but not denervation. A degradative response of cartilage of IL-1 beta was observed, and was enhanced by innervation. This model was useful for investigation of the articular response to acute inflammation and the influence of denervation in modulating this response.


Prognostic significance of infarction location in patients with recurrent myocardial infarction. SPRINT Study Group. Secondary Prevention Reinfarction Israel Nifedipine Trial.
This study assesses the impact of infarct location on immediate (in-hospital) and 1- and 5-year mortality among patients with reinfarction during the year following discharge from the initial episode of myocardial infarction. The analysis included 192 patients with a second myocardial infarction who were compared in four infarction location groups. The in-hospital mortality associated with reinfarction was higher in patients with a second anterior (32%) than with a second inferior (18%) location, irrespective of the first infarction location (p = 0.03). At 5 years of follow-up, the mortality (65%) tended to be higher in patients with a first anterior-second anterior infarction as compared with patients with all other combinations of location.


Vasopressin, theophylline, PGE2, and indomethacin on active Na transport in frog skin: studies with microelectrodes.
Active transepithelial Na transport in frog skin is influenced by vasopressin, theophylline, indomethacin, and PGE2. During stimulation or inhibition of the short-circuit current, the transapical membrane voltage of short-circuited skins was recorded using an intracellular microelectrode. The microelectrode also permitted determination of the fractional resistance of the apical barrier of the cells (fRo) and the E'1 (transepithelial voltage at which the apical membrane voltage is zero). Analysis of the data according to an electrical model proposed previously indicated that changes of ISC were mediated primarily via changes of the slope resistance Rfo (Vo negative) of the apical barrier of the cells with little or no effect on the Thévenin emf or resistance of the basolateral membranes. These data are in accordance with previous observations that ADH had no effect on the ENa and are discussed in relation to the origin of the ENa at the basolateral membranes of the epithelial cells.


Clinical presentation of candidaemia in elderly patients: experience in a single institution.
To analyse the clinical presentation of candidaemia in elderly patients. A comparison of clinical presentation of candidaemia cases was carried out in a Spanish tertiary hospital between January 2010 and September 2015. Forty-five cases (32%) corresponded to elderly patients (≥ 75 years) and 95 cases (68%) to non-elderly patients (16-74 years). A higher proportion of elderly patients presented solid tumour (51% versus 32%, p=0.026) and a lower proportion had undergone solid or hematopoietic transplantation (0% versus 28%, p<0.001). Fewer elderly patients (16 patients, 36%) had a central venous line inserted than non-elderly patients (81 patients, 85%, p<0.001). Isolation of Candida parapsilosis was significantly lower among elderly (13.3%) than among non-elderly patients (32%, p=0.015). Fundoscopy was carried out in 20 elderly (44%) and in 64 younger patients (67%, p=0.009). The proportion of patients who underwent echocardiography was similar in both groups (56% vs 66%, respectively; p=0.218). Adequate antifungal treatment within the first 48 hours was administered in16 elderly patients (36%) and 58 younger patients (61%, p=0.005). Catheter removal was carried out in 9 elderly patients (68.1%) and in 40 non-elderly patients (49%, p=0.544). Mortality was higher among elderly patients (55.6%) than non-elderly patients (36.8%; p=0.037). Elderly patients account for a substantial proportion of patients suffering from candidaemia in recent years. The clinical management of these patients was less appropriate than in younger patients with respect to fundus examination and the prescription of appropriate antifungal treatment. Mortality in elderly patients was higher than in younger patients.


Measuring auditory event-related potentials at the external ear canal: A demonstrative study using a new electrode and error-feedback paradigm.
Although ear canal electroencephalogram (EEG) recording has received interest from basic and applied research communities, evidence on how it can be implemented in practice is limited. The present study involving eight male participants including the authors presents the utility of our ear canal electrode and method by demonstrating both comparability of ear canal EEG to those at nearby sites and distinctiveness that ear canal event-related potentials (ERPs) could have. For this purpose, we used the balanced noncephalic electrode reference and an experimental paradigm with an error-feedback sound. Clear auditory ERPs were detected at the ear canal sites with a sufficiently low noise level comparable with those at conventional sites. The N1c, a temporal maximum subcomponent, spread over the bilateral temporal sites, including the ear canals and earlobes. While consecutive signals are generally highly similar between the ear canal and the earlobe, the N1c was larger at the ear canal than the earlobe, as demonstrated by the conventional frequentist and the hierarchical Bayesian modelling approaches. Although an evident caveat is that our sample was limited in terms of size and sex, the general capability indicates that the structure of our ear canal electrode provides EEG measurement that can be used in basic and applied settings. Our experimental method can also be an ERP-based test that conveniently assesses the capability of existing and future ear canal electrodes. The distinctive nature of the ERPs to the error-feedback sound may be utilized to examine the basic aspects of auditory ERPs and to test the processes involved in feedback-guided behaviour of participants.


T(h)2 cytokine dependence of IgD production by normal human B cells.
IgD is a minor component of serum Ig and the control of IgD secretion is virtually unknown. We measured concentrations of IgD (and IgE and IgM as controls) in culture supernatants of peripheral blood mononuclear cells (PBMC) from 60 normal donors as well as mononuclear cells from 10 tonsils following culture in the absence or presence of CD40 mAb and cytokines. Low levels of IgD were measured in cultures of PBMC, either unstimulated or stimulated by anti-CD40 antibodies. IL-4 and IL-10 significantly increased IgD production by CD40 mAb-stimulated cells in the majority of normal subjects studied, whereas in a limited number of individuals, spontaneous IgD production was either low or high, but with no increase upon stimulation. Spontaneous IgD production by tonsil-derived mononuclear cells was higher than by PBMC and increased after CD40 stimulation and even more in the presence of IL-10, but not IL-4. IL-2 and IFN-gamma exerted a dose-dependent inhibition on spontaneous as well as CD40- and cytokine-induced IgD production by PBMC, but not by tonsil mononuclear cells. Activation by IL-4 of CD40-stimulated purified B cells from tonsil and PBMC, and by IL-10 of tonsil B cells increased IgD production, whereas IL-2 and IFN-gamma had no detectable inhibitory effect. This suggests that accessory cells indirectly regulate IgD synthesis. IgD production induced in PBMC by IL-4 or IL-10 appeared to result from an active synthesis, and correlated with an increase in the number of IgD-containing plasma cells as demonstrated by immunofluorescence and increased expression of secreted IgD transcripts. These findings suggest that IgD production by normal peripheral blood human B cells is regulated positively by T(h)2 cytokines and negatively by T(h)1 cytokines.


The activated clotting time in cardiac surgery: should Celite or kaolin be used?
Both kaolin- and Celite-activated clotting times (ACT) are used to guide anticoagulation during cardiopulmonary bypass. It is unknown whether these methods lead to similar management procedures for anticoagulation in patients and are thus interchangeable in terms of bias, precision and variability. We randomized 97 patients undergoing coronary artery bypass grafting or aortic valve replacement to either kaolin- or Celite-guided anticoagulation. The ACT was measured simultaneously with the other method. We administered 300 IU/kg heparin to obtain initial ACT values greater than 400 s and additional heparin in each group using the minimum value of duplicate measurements according to a predefined protocol. The primary end point was the total heparin dose and the number of heparin supplements. The total heparin dose per patient in the 48 Celite-guided patients was 35 271 ± 12 406 IU with 51 supplements and in the 49 kaolin-guided patients, 35 997 ± 11 540 IU ( P = 0.77) with 56 supplements ( P = 0.53). Postoperative thrombin generation time, fibrinolytic response time, chest tube loss and transfusion requirements were not different between the two groups. However, the methods differed in individual patients with regard to supplemental heparin ( P = 0.002). Bias between methods at baseline was +10.3%, Celite being higher, and changed to a value of -12.9% at 2 h bypass. The coefficient of variation at baseline for individual patients was 2.6 times larger with kaolin than with Celite ( P < 0.001). Correlation between ACT values at baseline was only 45%. Kaolin- and Celite-guided management of anticoagulation is clinically not different, but the methods are not interchangeable. www.trialregister.nl identifier 1738.


Anxiogenic doses of rapamycin prevent URB597-induced anti-stress effects in socially defeated mice.
Repeated exposure to psychosocial stress modulates the endocannabinoid system, particularly anandamide (AEA) signaling in brain regions associated with emotional distress. The mTOR protein regulates various neuroplastic processes in the brain disrupted by stress, including adult hippocampal neurogenesis. This kinase has been implicated in multiple effects of cannabinoid drugs and the anti-stress behavioral effects of psychoactive drugs. Therefore, our hypothesis is that enhancing AEA signaling via pharmacological inhibition of the fatty acid amide hydrolase (FAAH) enzyme induces an anti-stress behavioral effect through an mTOR-dependent mechanism. To test this hypothesis, male C57Bl6 mice were exposed to social defeat stress (SDS) for 7 days and received daily treatment with either vehicle or different doses of the FAAH inhibitor, URB597 (0.1; 0.3; 1 mg/Kg), alone or combined with rapamycin. The results suggested that URB597 induced an inverted U-shaped dose-response curve in mice subjected to SDS (with the intermediate dose of 0.3 mg/kg being anxiolytic, and the higher tested dose of 1 mg/Kg being anxiogenic). In a second independent experiment, rapamycin treatment induced an anxiogenic-like response in control mice. However, in the presence of rapamycin, the anxiolytic dose of URB597 treatment failed to reduce stress-induced anxiety behaviors in mice. SDS exposure altered the hippocampal expression of the mTOR scaffold protein Raptor. Furthermore, the anxiogenic dose of URB597 decreased the absolute number of migrating doublecortin (DCX)-positive cells in the dentate gyrus, suggesting an anti-anxiety effect independent of newly generated/immature neurons. Therefore, our results indicate that in mice exposed to repeated psychosocial stress, URB597 fails to counteract the anxiogenic-like response induced by the pharmacological dampening of mTOR signaling.


Prediction of cerebral ischaemia during carotid endarterectomy with preoperative CO2-reactivity studies and angiography.
The objective of this study was to assess the value of combining the preoperative CO2 cerebrovascular reactivity index (CO2RI) with carotid and cerebral angiography in predicting the risk of severe cerebral ischaemia (SCI) during carotid endarterectomy (CEA). Seventy-four consecutive patients scheduled for CEA underwent preoperative digital subtraction angiography and CO2-reactivity tests. During CEA, cerebral function monitor (CFM) was used to document cortical electrical activity, whilst transcranial Doppler measured the middle cerebral artery flow velocity (FV). A persistent fall in CFM voltage and/or a fall in FV > or = 60% on internal carotid artery (ICA) clamping were used as criteria for defining SCI. Complete data from 59 patients were obtained for final analysis. Twelve cases showed a fall in FV > or = 60%; 11 of these also showed a sustained fall in CFM voltage. Using logistic regression, the risk of SCI was found to be negatively associated with (1) contralateral CO2RI, (2) the percentage stenosis of the contralateral ICA, and (3) the difference between ipsilateral and contralateral CO2RI. Using these factors, a logistic regression model for predicting the risk of SCI was established which provided a sensitivity of 75% and specificity of 100%. The risk of SCI during CEA was related to the contralateral ICA stenosis and the CO2RI of both cerebral hemispheres. This information may assist in presurgical planning and help to select asymptomatic carotid lesions for surgery.


Regulation of Wilms' tumor gene expression by nerve growth factor and follicle-stimulating hormone in the immature mouse ovary.
This study investigated the regulation of Wilms' tumor gene (WT1) in the ovary by nerve growth factor and FSH to better understand signals that initiate early follicular growth. Nerve growth factor showed a direct stimulatory effect on endogenous expression of WT1, whereas FSH attenuated basal and nerve growth factor-stimulated WT1 protein expression, which most likely depended on FSH responsiveness according to the follicle growth stage.


New African American life tables from 1935-1940 to 1985-1990.
New life tables for African Americans are presented from 1935 through 1990. They are based on a historical series of vital statistics data on deaths that have been corrected for age misreporting, on reconstructed population counts that have been adjusted for census underenumeration, and on births that have been corrected for underregistration. The new life tables show rapid mortality declines for both African American males and females from 1935 to 1950, and relatively steady reductions thereafter for females. The smaller declines in male mortality in young adulthood and middle age since the 1950s have led to exceptionally high ratios of male to female mortality at these ages. Corrections for census undercounts lead to higher values of life expectancy than in official life tables, but to less improvement over time. Official estimates of life expectancy at age 65 appear to be about 10% too high around 1940 but only about 1.5% too high in the late 1980s.


Repeated exposures to enzyme-generated oxidants cause alveolitis, epithelial hyperplasia, and fibrosis in hamsters.
Oxidant-mediated epithelial injury and repair processes may promote the development of pulmonary fibrosis. The authors examined this hypothesis by inducing oxidant injury in hamsters with intratracheally instilled mixtures of glucose, glucose oxidase (GO) and lactoperoxidase at weekly intervals. Solutions containing denatured GO (DE) served as a control treatment. One and six days after each treatment, anesthetized animals were sacrificed and lavaged, and their lungs and plasma were preserved for further study. Although DE-treatment consistently evoked a transient, neutrophil-rich inflammatory response, no significant biochemical or morphologic changes were detected at the ensuing 6-day time points. In contrast, repeated GO treatments prolonged inflammation and injured the alveolar epithelium, evidenced by significantly greater levels of neutrophils and macrophages in bronchoalveolar lavage fluid (BALF) and increased BALF levels of protein, beta-glucuronidase and lactic dehydrogenase activities. Active GO also altered BALF lymphocytes and monocytes, but no discernable pattern emerged. Fibrotic, consolidated parenchyma appeared after the second and third GO exposures, coinciding with increased levels of total collagen, prolyl hydroxylase activity, and anti-oxidant enzyme activities. Although alveolitis and type II cell hyperplasia were observed after the initial treatment, polyplike nodules covered by hyperplastic, undifferentiated epithelium were evident after the third treatment. After each exposure, GO-treated animals had larger volumes of parenchymal lesion than DE-treated hamsters. These data indicate that normal alveolar epithelial repair processes were greatly disrupted by repeated oxidant injury and suggest that repeated and/or continued epithelial injury may contribute to the development of pulmonary fibrosis.


Functional characterization and expression analysis of a gene, OsENT2, encoding an equilibrative nucleoside transporter in rice suggest a function in cytokinin transport.
We identified four genes for potential equilibrative nucleoside transporters (ENTs) from rice (Oryza sativa; designated OsENT1 through OsENT4). Growth analysis of budding yeast (Saccharomyces cerevisiae) cells expressing OsENTs showed that OsENT2 transported adenosine and uridine with high affinity (adenosine, K(m) = 3.0 microm; uridine, K(m) = 0.7 microm). Purine or pyrimidine nucleosides and 2'-deoxynucleosides strongly inhibited adenosine transport via OsENT2, suggesting that OsENT2 possesses broad substrate specificity. OsENT2-mediated adenosine transport was resistant to the typical inhibitors of mammalian ENTs, nitrobenzylmercaptopurine ribonucleoside, dilazep, and dipyridamole. The transport activity was maximal at pH 5.0 and decreased slightly at lower as well as higher pH. In competition experiments with various cytokinins, adenosine transport by OsENT2 was inhibited by isopentenyladenine riboside (iPR). Direct measurements with radiolabeled cytokinins demonstrated that OsENT2 mediated uptake of iPR (K(m) = 32 microm) and trans-zeatin riboside (K(m) = 660 microm), suggesting that OsENT2 participates in iPR transport in planta. In mature plants, OsENT2 was predominantly expressed in roots. The OsENT2 promoter drove the expression of the beta-glucuronidase reporter gene in the scutellum during germination and in vascular tissues in germinated plants, suggesting a participation of OsENT2 in the retrieval of endosperm-derived nucleosides by the germinating embryo and in the long-distance transport of nucleosides in growing plants, respectively.


Identification of candidate genes controlling chilling tolerance of rice in the cold region at the booting stage by BSA-Seq and RNA-Seq.
Rice is sensitive to low temperatures, specifically at the booting stage. Chilling tolerance of rice is a quantitative trait loci that is governed by multiple genes, and thus, its precise identification through the conventional methods is an arduous task. In this study, we investigated the candidate genes related to chilling tolerance at the booting stage of rice. The F2 population was derived from Longjing25 (chilling-tolerant) and Longjing11 (chilling-sensitive) cross. Two bulked segregant analysis pools were constructed. A 0.82 Mb region containing 98 annotated genes on chromosomes 6 and 9 was recognized as the candidate region associated with chilling tolerance of rice at the booting stage. Transcriptomic analysis of Longjing25 and Longjing11 revealed 50 differentially expressed genes (DEGs) on the candidate intervals. KEGG pathway enrichment analysis of DEGs was performed. Nine pathways were found to be enriched, which contained 10 DEGs. A total of four genes had different expression patterns or levels between Longjing25 and Longjing11. Four out of the 10 DEGs were considered as potential candidate genes for chilling tolerance. This study will assist in the cloning of the candidate genes responsible for chilling tolerance and molecular breeding of rice for the development of chilling-tolerant rice varieties.


Cigarette smoking and serious psychological distress: a population-based study of California adults.
This study examines differences in smoking behaviors between adults with and without serious psychological distress (SPD) in California, which has the longest running comprehensive tobacco control program in the world. Cross-sectional data from the 2007 California Health Interview Survey on 50,880 noninstitutionalized adults were used to analyze smoking prevalence, cigarette consumption, and quit ratio. Persons with SPD were identified using the K6 scale, a clinically validated psychological screening instrument. About 3.8% of California adults screened positive for SPD in the past 30 days (acute SPD) and an additional 4.8% screened positive for SPD in the past 2-12 months (recent SPD). Persons with SPD were more likely to be current smokers than those without SPD (adjusted odds ratios [AOR] = 2.54, 95% CI = 2.02-3.19 for acute SPD and AOR = 2.20, 95% CI = 1.79-2.71 for recent SPD). Current smokers with acute SPD were more likely to smoke ≥20 cigarettes daily than those without SPD (AOR = 1.59, 95% CI = 1.06-2.39). The quit rate was lower among ever-smokers with acute (AOR = 0.46, 95% CI = 0.35-0.62) or recent SPD (AOR = 0.55, 95% CI = 0.42-0.71) than those without SPD. While persons with acute or recent SPD comprised 8.6% of adults, they consumed 19.2% of all cigarettes in California. In California, adults with SPD were more likely to be current smokers and to smoke heavily and less likely to quit than those without SPD. The findings underscore the need for effective smoking cessation strategies targeting this group.


Solid nanotubes comprising alpha-Fe2O3 nanoparticles prepared from ferritin protein.
Solid nanotubes comprising alpha-Fe2O3 nanoparticles were prepared from iron-storage protein ferritin. Their structure, magnetic properties, and photocatalytic activities were characterized. The initial ferritin nanotube precursors were fabricated using alternating layer-by-layer depositions of poly-L-arginine (PLA) and ferritin into a track-etched polycarbonate membrane (pore diameter, 400 nm) with subsequent dissolution of the template. The obtained uniform cylinders of (PLA/ferritin)3 (outer diameter, 410 +/- 14 nm) were calcinated at 500 degrees C under air, yielding reddish-brown iron oxide nanotubes. The one-dimensional hollow structure remained perfect, but its diameter, wall thickness, and maximum length were markedly diminished. Disappearance of the protein shell and the PLA layers were confirmed using IR and EDX spectroscopy. Subsequent SEM, TEM, and XPS measurements showed that the tubular walls comprise fine alpha-Fe2O3 nanoparticles with a 5 nm diameter. These alpha-Fe2O3 nanotubes demonstrated superparamagnetic properties with a blocking temperature of 37 K and efficient photocatalytic activity for degradation of 4-chlorophenol.


The origin of post-injury neointimal cells in the rat balloon injury model.
The origin of post-injury neointimal cells is still a matter of debate. This study aims to determine the anatomic source of neointimal cells in one of the most important animal models for the study of vascular stenosis in response to injury, the rat balloon injury model. Chimeric rats were generated by rescuing lethally irradiated animals with green fluorescent protein (GFP)(+) bone marrow (BM) cells from transgenic rats. Neointimal formation was induced in the right iliac artery of these animals using a balloon angioplasty catheter. Injured and non-injured contra-lateral arteries were harvested at 7, 14, and 30 days post-surgery. BM-derived monocytes/macrophages (CD68(+) GFP(+)) were abundant in the media and adventitia of injured vessels harvested at 7 days as determined by immunofluorescence and confocal microscopy. The number of GFP(+) cells declined in the vascular wall with time. Post-injury neointimal cells were mostly GFP(-)/smooth muscle actin (SMA)(+), which indicated that those cells originated in the recipient. Only a few neointimal cells seemed to come from circulating progenitors (GFP(+) SMA(+), 2.34% +/- 1.61). The vascular origin of cells in the neointima was further confirmed by transplanting injured GFP arteries into wild-type recipients. In these grafts, 94.23 +/- 0.44% of medial and 92.95 +/- 19.34% of neointimal cells were GFP(+) SMA(+). Finally, we tested the capacity of vascular smooth muscle cells (VSMC) to migrate through the vascular wall using a novel in vivo assay. As expected, VSMC migrated and populated the neointima only in response to injury. Our results suggest that neointimal cells in the rat balloon injury model mostly derive from pre-existing vascular cells and that only a small population of those cells come from BM-derived progenitors.


[Study of the effect of reality orientation on a group of psychogeriatric patients].
In 22 mildly deteriorated elderly patients the total score on a reality orientation questionnaire improved after 3 months ROT. This increased knowledge on basic facts was ascertained in both groups: patients who attended the ROT-class and patients who participated in the ROT-discussion-group. It became clear though that the effect of the ROT was limited to those items of the questionnaire that had been offered to the patients. The knowledge of items which had not been offered in the ROT remained unchanged. ROT had no generalized effect on the verbal orientation. It only had a specific effect on trained items of information.


Malaria theranostics using hemozoin-generated vapor nanobubbles.
Malaria remains a widespread and deadly infectious human disease, with increasing diagnostic and therapeutic challenges due to the drug resistance and aggressiveness of malaria infection. Early detection and innovative approaches for parasite destruction are needed. The high optical absorbance and nano-size of hemozoin crystals have been exploited to detect and mechanically destroy the malaria parasite in a single theranostic procedure. Transient vapor nanobubbles are generated around hemozoin crystals in malaria parasites in infected erythrocytes in response to a single short laser pulse. Optical scattering signals of the nanobubble report the presence of the malaria parasite. The mechanical impact of the same nanobubble physically destroys the parasite in nanoseconds in a drug-free manner. Laser-induced nanobubble treatment of human blood in vitro results in destruction of up to 95% of parasites after a single procedure, and delivers an 8-fold better parasiticidal efficacy compared to standard chloroquine drug treatment. The mechanism of destruction is highly selective for malaria infected red cells and does not harm neighboring, uninfected erythrocytes. Thus, laser pulse-induced vapor nanobubble generation around hemozoin supports both rapid and highly specific detection and destruction of malaria parasites in one theranostic procedure.


A comparative pharmacokinetic study of a fixed dose combination for essential hypertensive patients: a randomized crossover study in healthy human volunteers.
This study was aimed to investigate the relative bioavailability of fixed-dose-combination (FDC) product of amlodipine, telmisartan and hydrochlorothiazide with individual marketed products in healthy male volunteers. Control of blood pressure with fixed dose combination of the above drugs acting through different mechanism have a benefit of convenient dosing in terms of compliance, lower the dose and subsequently reduce the side effects. The authors investigated the relative bioavailability under a fasting state of the 3 drugs in a randomized, open-label, 2-treatment, 2-period, 2-sequence, crossover bioequivalence study with a washout period of 21 days. Plasma concentration of the analytes were assayed in timed samples with a simple, highly sensitive and rapid validated method using HPLC coupled to tandem mass spectrometry that had a lower limit of quantification of 1 ng/mL for all the 3 components. Test and reference formulations gave a mean Cmax of 5.234±0.914 ng/mL and 4.991±0.563 ng/mL, 108.839±13.601 ng/mL and 114.783±12.315 ng/mL and 97.814±10.779 ng/mL and 93.731±10.018 ng/mL for amlodipine, telmisartan and hydrochlorothiazide respectively. The AUC0-t of amlodipine, telmisartan and hydrochlorothiazide was 161.484 ng.h/mL, 1 917.644 ng.h/mL and 822.847 ng.h/mL for test formulation and 162.108 ng.h/mL, 2 014.764 ng.h/mL and 829.323 ng.h/mL for reference in the fasting state. The 90% confidence intervals for the test/reference ratio of the pharmacokinetic parameters in fasting state (mean Cmax, AUC0-t, and AUC0-∞) were within the acceptable range of 80.00-125.00. Thus, these findings clearly indicate that the FDC product is bioequivalent with the individual marketed products in terms of rate and extent of drug absorption and is well tolerated with no significant adverse reactions.


Safety evaluation of standardized Ayurvedic formulation-Panchvalkala, by subacute toxicity study.
Panchvalkala is a conventional Ayurvedic medicine used as a douche in gynecological disorders such as leucorrhea, infertility, and endometriosis. Recently, we have reported the anticancer activity of Panchvalkala aqueous extract (PVaq) in cervical cancer cell lines, SiHa (HPV16+), HeLa (HPV18+), and mouse papilloma models. Here, we have evaluated the safety of the aqueous extract of Ayurvedic formulation, Panchvalkala (PVaq), in Swiss albino mice by performing subacute toxicity study. Male and female Swiss albino mice (n = 5/sex/group) were gavaged orally with different doses of PVaq for 28 consecutive days. The mice were distributed into six groups: I (vehicle control), II (vehicle control reversal), III (PVaq 250 mg/kg), IV (PVaq 500 mg/kg), V (1000 mg/kg) and VI (1000 mg/kg high dose reversal). Animals were observed periodically to record any clinical signs of toxicity or mortality. After completion of treatment and recovery periods, animals were evaluated for the effect of PVaq on urine parameters, followed by hematological and biochemical parameters. Animals were sacrificed on day 29 for gross observation of vital organs and to study their histopathology. Reversal groups were maintained for further 14 days to observe any delayed onset of toxic side effects or reversal of toxicity, followed by sacrificing the mice on day 43. In the subacute toxicity study, PVaq did not show any significant change in food, water consumption, and body weights. There were no significant alterations in hematology, biochemistry, urine parameters, and histopathology of the analyzed tissues (brain, heart, liver, lung, spleen, thymus, kidney, epididymis/ovaries, and testis/uterus). The parameters were comparable to their respective controls in both the female as well as the male mice groups. Upon macroscopic and microscopic observation of vital organs, no abnormality was detected compared to the respective control groups. The subacute toxicity study demonstrated that oral administration of PVaq was safe in female and male Swiss albino mice.


[Relese of Ca2+ from mitochondria after mitochondrial membrane depolarisation].
With the aid of specific inhibitors of Ca(2+)-uniporter (ruthenium red) and mitochondrial permeability transition pore, PTP (cyclosporine A) it is shown that PTP opening takes place after loading the rat liver mitochondria with calcium and depolarisation of mitochondrial membrane with protonophore (carbonyl cyanide m-chlorophenyl hydrazone, CCCP), and the pore opening accounts for accelerated efflux of calcium from mitochondrial matrix as well as availability of "rapid" component of two-exponential kinetic curve of Ca(2+)-efflux. An analysis of kinetic data of Ca2+ transport after membrane depolarisation also confirms our earlier observations that time frame of the pore open state is restricted, and membrane integrity is restored before all the calcium load is delivered into incubation medium. The absence of additivity between the shares of Ca(2+)-uniporter and PTP in Ca(2+)-transport is observed, and conclusion is made that partial share of PTP in calcium transport is not a constant, but a variable constituent which is diminished to zero as soon as the Ca(2+)-uniporter activity reaches its maximum after the abolition of membrane potential with CCCP. Based on some observations, it is supposed also that PTP inactivation takes place during calcium translocation across the mitochondrial membrane, which could account for limited release of Ca2+ from mitochondrial matrix through the pore itself as well as relatively narrow limits of the pore open state in comparison with time scale of complete cation release from depolarised mitochondria.


Risk Factors for the Development of Esophagorespiratory Fistula in Esophageal Cancer.
The development of esophagorespiratory fistula (ERF) in esophageal cancer (EC) is a devastating complication, leading to poor survival rates and low quality of life. Goal of this study was to identify risk factors leading to fistula formation in esophageal cancer. We identified 47 patients with malignant ERF formation in EC in a period of 10 years. Clinical characteristics were compared by univariable analysis to 47 randomly selected patients with EC, but without ERF. A case-control study was conducted for patients with squamous cell carcinoma (SCC) and ERF matching in a 1:2 fashion for primary tumor localization. Identifiable risk factors in EC patients were histology of SCC (P-value < 0.001), former or current smoking status (P = 0.002) and primary tumor localization in the proximal esophagus (P < 0.001). The "hot spot" for ERF formation was tumor growth 20-25cm distal to dental arch. An additional risk factor in SCC patients was age. Patients with ERF formation in SCC were younger than patients without ERF (median 63 vs. 67 years, P = 0.02). No difference in the rate of fistula formation was seen between esophagectomy and definitive chemoradiation, but the latter developed ERF earlier in the course of the disease (237 vs. 596.5 days, P = 0.01). Patients with proximal SCC of the esophagus and a smoking history, as well as young patients with SCC should be closely monitored for ERF formation.


Hemolytic factors in Schistosoma japonicum eggs.
Extracts of Schistosoma japonicum eggs were found to exhibit hemolytic activity on erythrocytes of various species. The hemolytic reaction took place more rapidly at 37 degrees C than at 4 degrees C and did not require divalent cations. The degree of hemolysis was dependent on the concentration of the egg extracts. The hemolytic factors seemed to be lipid in nature because of being heat-stable and soluble in chloroform solvent. Further analysis by means of thin-layer chromatography showed that hemolytic activity in the extracts was due to free fatty acids. Analysis by gas chromatography revealed that fatty acids in the egg extracts consist of myristic, palmitic, palmitoleic, stearic, oleic, linoleic, linolenic, and arachidonic acids. Among them, based on the experiments with commercially available fatty acids, arachidonic acid was the most intensively hemolytic, followed by linoleic, linolenic, oleic, palmitoleic, and myristic acids in that order, whereas palmitic acid showed weak activity only in the presence of divalent cations. From the estimated calculation, it was assumed that 1 mg of protein of the egg extracts contains as much as 0.22 mg of free fatty acids.


Climate change: Overview of risks to pregnant persons and their offspring.
Climate change is one of the greatest challenges confronting humanity. Pregnant persons, their unborn children, and offspring are particularly vulnerable, as evidenced by adverse perinatal outcomes and increased rates of childhood illnesses. Environmental inequities compound the problem of maternal health inequities, and have given rise to the environmental justice movement. The International Federation of Gynecology and Obstetrics and other major medical societies have worked to heighten awareness and address the deleterious health effects of climate change and toxic environmental exposures. As part of routine prenatal, neonatal, and pediatric care, neonatal-perinatal care providers should incorporate discussions with their patients and families on potential harms and also identify actions to mitigate climate change effects on their health. This article provides clinicians with an overview of how climate change affects their patients, practical guidance in caring for them, and a frame setting of the articles to follow. Clinicians have a critical role to play, and the time to act is now.


Hierarchical thermoplastic rippled nanostructures regulate Schwann cell adhesion, morphology and spatial organization.
Periodic ripples are a variety of anisotropic nanostructures that can be realized by ion beam irradiation on a wide range of solid surfaces. Only a few authors have investigated these surfaces for tuning the response of biological systems, probably because it is challenging to directly produce them in materials that well sustain long-term cellular cultures. Here, hierarchical rippled nanotopographies with a lateral periodicity of ∼300 nm are produced from a gold-irradiated germanium mold in polyethylene terephthalate (PET), a biocompatible polymer approved by the US Food and Drug Administration for clinical applications, by a novel three-step embossing process. The effects of nano-ripples on Schwann Cells (SCs) are studied in view of their possible use for nerve-repair applications. The data demonstrate that nano-ripples can enhance short-term SC adhesion and proliferation (3-24 h after seeding), drive their actin cytoskeleton spatial organization and sustain long-term cell growth. Notably, SCs are oriented perpendicularly with respect to the nanopattern lines. These results provide information about the possible use of hierarchical nano-rippled elements for nerve-regeneration protocols.


Life after the loss: protocol for a Danish longitudinal follow-up study unfolding life and grief after the death of a child during pregnancy from gestational week 14, during birth or in the first 4 weeks of life.
After the death of a child during pregnancy, birth or in the neonatal period, parents often experience feelings of guilt, disenfranchisement, feelings of betrayal by one's own body and envy of others. Such bereavement results in high rates of distress: psychologically, emotionally, physiologically and existentially. These data are collected using a national, longitudinal cohort to assess grief in mothers and their partners after the death of a child during pregnancy, birth or in the neonatal period. Our aim is to achieve a general description of grief, emotional health, and existential values after pregnancy or perinatal death in a Danish population. The cohort comprises mothers and their partners in Denmark who lost a child during pregnancy from gestational week 14, during birth or in the neonatal period (4 weeks post partum). We began data collection in 2015 and plan to continue until 2024. The aim is to include 5000 participants by 2024, generating the largest cohort in the field to date. Parents are invited to participate at the time of hospital discharge or via the Patient Associations homepage. Data are collected using web-based questionnaires distributed at 1-2, 7 and 13 months after the loss. Sociodemographic and obstetric variables are collected. Validated psychometric measures covering attachment, continuing bonds, post-traumatic stress, prolonged grief, perinatal grief and existential values were chosen to reach our aim. The study was approved by The Danish National Data Protection Agency (no. 18/15684, 7 October 2014). The results will be disseminated in peer-reviewed and professional journals as well as in layman magazines, lectures and radio broadcasts.


Gene flow from weedy red rice (Oryza sativa L.) to cultivated rice and fitness of hybrids.
Gene transfer from weeds to crops could produce weedy individuals that might impact upon the evolutionary dynamics of weedy populations, the persistence of escaped genes in agroecosystems and approaches to weed management and containment of transgenic crops. The present aim was to quantify the gene flowrate from weedy red rice to cultivated rice, and evaluate the morphology, phenology and fecundity of resulting hybrids. Field experiments were conducted at Stuttgart and Rohwer, Arkansas, USA. Twelve red rice accessions and an imazethapyr-resistant rice (Imi-R; Clearfield) were used. Hybrids between Imi-R rice x red rice were 138-150 cm tall and flowered 1-5 days later than the rice parent, regardless of the red rice parent. Hybrids produced 20-50% more seed than the rice parent, but had equivalent seed production to the majority of red rice parents. Seeds of all hybrids were red, pubescent and dehisced at maturity. For the majority of hybrids, seed germination was higher than that of the red rice parent. The gene flowrate from red rice to rice was 0.01-0.2% and differed by red rice biotype. The hybrids had higher fecundity and potential competitive ability than the rice parent, and in some cases also the red rice parent. Red rice plants are vectors of gene flow back to cultivated rice and other weedy populations. The progeny of red rice hybrids from cultivated rice mother plants have higher chances of persistence than those from red rice mother plants. Gene flow mitigation strategies should consider this scenario.


Overview of the different methods for RNA preparation in COVID-19 diagnosis process during the pandemic.
The COVID-19 pandemic brought to light the impact of a widespread disease on various aspects of human relationships, communities, and economies. One notable consequence was the increased demand for diagnostic kits, laboratory reagents, and personal health equipment. This surge in testing capacity worldwide led to shortages in the supply of essential items, including RNA extraction kits, which are crucial for detecting COVID-19 infections. To address this scarcity, researchers have proposed alternative and cost-effective strategies for RNA extraction, utilizing both chemical and physical solutions and extraction-free methods. These approaches aim to alleviate the challenges associated with the overwhelming number of tests being conducted in laboratories. The purpose of this review is intends to provide a comprehensive summary of the various kit-free RNA extraction methods available for COVID-19 diagnosis during the pandemic.


A prospective study evaluating the role of donor-specific anti-endothelial crossmatch (XM-ONE assay) in predicting living donor kidney transplant outcome.
Anti-endothelial cell antibodies (AECAs) may play a role in allograft rejection. We prospectively tested 150 consecutive living donor kidney transplant recipients, with transplants performed at Northwestern Memorial Hospital between January and December 2010, using the donor-specific endothelial (XM-ONE) crossmatch. 88/150 Patients received standard of care (SOC) immunosuppression and analyzed separately, in addition to the complete study cohort. Patients were followed for one year and XM-ONE results were analyzed in relation to occurrence of acute rejection, proteinuria, serum creatinine levels, and biopsy proven fibrosis. No correlation was found between XM-ONE results and protocol or "for-cause" biopsy proven acute rejection or vasculopathy at 12 months. When IgG+ and IgM+ results of the XM-ONE assay were combined, a correlation with proteinuria at 12 months was observed (p=0.047). Although IgG+XM-ONE results were associated with significantly higher creatinine at 6 months (p=0.018), significance was lost at 12 months. Conversely, patients with an IgM+XM-ONE crossmatch had significantly lower creatinine at 1 month (p=0.019), 3 months (p=0.0045), and 6 months (p=0.038) post-transplant, but lost statistical significance at 12 months (p=0.67) post-transplant. In summary, the presence of AECAs as determined by a positive XM-ONE result was not predictive of overall poorer graft outcome after one year in our center.


Obesity, bariatric surgery, and bone.
A greater number of morbidly obese individuals are undergoing bariatric surgery, expanding the postbariatric surgery population. Both obesity and bariatric surgery are accompanied by a variety of vitamin and mineral deficiencies, with some affecting bone health. Currently, there is no consensus regarding bone health evaluation and its management in obese and postbariatric population. This review will focus on nutritional deficiencies in obese and postbariatric surgical patients, with a special focus on bone health. The latest findings on the complicated relationship between adipose tissue and bone will be discussed. And the presumed protective effect of obesity on osteoporosis will be evaluated. Nutritional deficiencies in postbariatric patients already exist prior to the surgery. Therefore, a comprehensive nutritional evaluation prior to bariatric surgery is imperative, to prevent further decline of the already deficient nutrients. Nutrients such as calcium and vitamin D have an effect on bone health. Therefore, monitoring their level is important in preventing bone loss. Taking a multivitamin with minerals postbariatric surgery is a standard of practice.


Structured primary care for type 2 diabetes has positive effects on clinical outcomes.
Patients with type 2 diabetes have an increased risk of developing microvascular and macrovascular complications. In routine diabetes care an adequate reduction of risk factors for these complications is often not achieved. The aim of the study was to evaluate the effects of structured diabetes care on clinical outcomes of patients with type 2 diabetes in primary care. We performed a quasi-experimental study on the effects of structured care consisting of organizational and educational components (n = 581) compared with care-as-usual (n = 152). We assessed clinical outcomes of HbA1c, blood pressure, cholesterol, creatinine and body mass index, at baseline and after 1 year. The long-term effects in the structured care group were determined after another 2 years. Structured care led to improvement in HbA1c and long-term improvements in blood pressure and cholesterol compared with care-as-usual. After 1 year, the percentage of patients who did not deteriorate was higher in the structured care group, again for HbA1c, diastolic blood pressure, low-density lipoprotein cholesterol and body mass index. Structured diabetes care consisting of multiple components has a positive effect on clinical outcomes compared with care-as-usual. Our findings support its further implementation in order to reduce complications in type 2 diabetes patients.


Transthyretin Val71Ala mutation in a Dutch family with familial amyloidotic polyneuropathy.
A Dutch family with familial amyloidotic polyneuropathy associated with the transthyretin mutation Val71Ala is described. This is the third reported family with this mutation, causing at the protein level an unstable TTR monomer and at the clinical level progressive wasting, polyneuropathy, autonomic dysfunction and vitreous opacities.