Improving culture initiation of mature oak shoots through use of silver thiosulfate.
Of the approximately 430 species of oaks (Quercus spp.) that have been assessed, 31% are threatened with extinction and in need of safeguarding. However, oak seeds cannot be seed banked, and thus rely on alternative strategies such as in vitro culture for ex situ conservation. One challenge to this approach is low culture initiation rates. Our objective was to identify factors that may improve the establishment of shoot cultures in vitro using new growth collected from mature trees. Shoot cuttings were harvested from individuals of five different oak species (Q. alba, Q. bicolor, Q. macrocarpa, Q. muehlenbergii, and Q. palustris). Shoots were cultured onto medium with or without 50 µM silver thiosulfate (STS), a known inhibitor of the stress hormone ethylene. Cultures were grown for one month, at which point shoots were assessed for survival. Shoot survival was significantly greater in shoots cultured on medium containing STS compared to the control group, with the overall survival rate increasing from 65% to 73%. Increasing the survival rate of newly established cultures is important in ensuring that material collected from endangered species has the best chance for survival, which is critical for successful ex situ conservation.


Effect of a Bicyclic Pyrimidine Derivative (KRP-103), a Novel Selective Tachykinin NK1 Receptor Antagonist, on Bladder Function in Guinea Pigs.
We evaluated the pharmacological characteristics of KRP-103, chemically named as ((2-(4-acetylpiperazin-1-yl)-6-[3,5-bis(trifluorometheyl)-phenylmethyl]-4- (2-methylphenyl)-6,7,8,9-tetrahydro-5H-pyrimido[4,5-b][1,5]oxazocin-5-one), and its effects on lower urinary tract function in guinea pigs. In radioligand binding assay, KRP-103 showed higher selectivity for human NK1 receptor (hNK1R) than for human NK2 receptor (hNK2R) and human NK3 receptor (hNK3R) (Ki for hNK1R, hNK2R and hNK3R=0.126 nM, > 10 000 nM and > 10 000 nM). In distention-induced rhythmic bladder contractions (RBCs) in urethane-anesthetized guinea pigs, intravenous administration of KRP-103 dose-dependently increased the shutdown time of RBCs and slightly decreased the amplitude of RBCs (only about 20%). In acutely spinalized guinea pig cystometory, intraduodenal administration of KRP-103 dose-dependently increased the effective bladder capacity with a minimum effective dose of 1 mg/kg. Furthermore, to clarify the site of action of KRP-103, we evaluated the inhibitory effects of KRP-103 on bladder contractions induced by electrical stimulation (ES) of the central or peripheral cut end of the pelvic nerve (PN). KRP-103 inhibited the bladder contractions induced by ES of the central cut end of PN but not those induced by ES of the peripheral cut end of PN. These results indicate that KRP-103 enhances bladder storage function by inhibiting sensory transmission from the bladder at the level of spinal cord without affecting bladder efferent function, suggesting that KRP-103 may be a suitable therapeutic drug for treatment of overactive bladder.


Accessory cell stimulation of T cell proliferation requires active antigen processing, Ia-restricted antigen presentation, and a separate nonspecific 2nd signal.
The roles of Ia+ accessory cells in H-2-restricted stimulation of antigen-specific T cell proliferation were explored in an in vitro model. L-glutamic acid60-L-alanine30-L-tyrosine10-(GAT) primed BALB/c nylon wool-passed T cells were depleted of Ia+ antigen-presenting cells (APC) by treatment with monoclonal anti-Ia antibody plus complement. Such cells failed to respond to soluble GAT, or to soluble GAT in the presence of phorbol myristic acetate (PMA), which is known to stimulate production of, or replace, IL-1 in vitro. Addition of gamma-irradiated syngeneic spleen cells reconstituted the response to soluble GAT, but addition of ultraviolet (UV) light-irradiated spleen cells did not, even in the presence of PMA. Preincubation of cells with GAT for 24 hr, followed by washing, then gamma irradiation, generated a cell population able to stimulate GAT-primed T cells to proliferate. The same pulsed cells exposed to UV irradiation failed to stimulate T cell responses unless PMA was added to the cultures. The relevant cells in this UV-irradiated population are Ia+. It is concluded that a finite period of time for interaction of metabolically intact APC with antigen is required before creation of an appropriate (Ia + antigen) signal recognized by the T cell. In addition to such Ia-restricted antigen presentation, however, a 2nd nonspecific signal, again requiring metabolically active APC for elaboration, is necessary for detectable T cell activation. These studies thus define 3 separable activities of APC during the process of H-2 restricted T cell activation.


Evaluation of early conception factor lateral flow test to determine nonpregnancy in dairy cattle.
The early conception factor (ECF) lateral flow test was evaluated for its ability to accurately determine nonpregnant status in dairy cattle. Results of 2 field trials involving 191 cows and 832 tests indicated the probability that a cow can be correctly diagnosed as nonpregnant by using the ECF test is only about 50%. Agreement of test results between milk and serum obtained from the same cow was 57.5%. The ECF test was not consistent in identifying nonpregnancy when the same cows were tested repeatedly over a period of 4 weeks. We conclude that the ECF lateral flow test does not accurately identify nonpregnancy in dairy cattle.


The hormone response element of the mouse mammary tumour virus DNA mediates the progestin and androgen induction of transcription in the proviral long terminal repeat region.
Mouse mammary tumour virus (MMTV) gene expression has been shown to be regulated by glucocorticoids. A hormone response element (HRE) located between -202 and -59 upstream of the start of transcription in the long terminal repeat (LTR) region of the proviral DNA is required for this induction. We have investigated the role played by the HRE in the induction of MMTV LTR transcription by other classes of steroid hormones. Chimaeric constructs containing the HRE and the authentic LTR promoter linked to an indicator gene or the HRE linked to an otherwise hormone insensitive promoter directing the transcription of an indicator gene, were transfected into the human mammary tumour cell line T47D. Transcription at the MMTV LTR promoter or at the previously hormone-insensitive promoter was induced by progestins and androgens but not by oestradiol in transfected cells that contained functional receptors for these hormones. These results identify the HRE as the cis-acting element that mediates the progestin and androgen induction of MMTV LTR transcription. The HRE is therefore a DNA element that is required not just for glucocorticoid but also for progesterone and androgen induction of MMTV LTR transcription.


Alterations in tRNA isoaccepting species during erythroid differentiation of the Friend leukemia cell.
The chromatographic profiles of isoaccepting tRNAs were analyzed at five time points during the 96 hr, dimethylsulfoxide induced, erythroid-like differentiation of Friend leukemia cells. Sixty-four isoaccepting species of tRNA for 16 amino acids were resolved by RPC-5 chromatography. The relative amounts of tRNAphe, tRNAile, and tRNAval species were maintained by the cells during differentiation; whereas the relative amounts of some of the isoacceptor tRNAs for the other 13 amino acids changed significantly. Fluctuations in amounts of isoacceptors occurred between 36 and 72 hr after addition of dimethysulfoxide, corresponding to globin mRNA appearance and hemoglobin synthesis, respectively. In most cases, thepredominant tRNA isoacceptors of uninduced cells were retained throughout differentiation. Notable exceptions were tRNA species for threonine, proline, and methionine. Some of the isoacceptors occurring in relatively smaller amounts were not expressed at all times. These changes possibly reflect the cell's functional adaptation of tRNA in differentiation for hemoglobin synthesis.


The feasibility, safety, and utility of vertebral wedge osteotomies for the fusionless treatment of paralytic scoliosis.
Before-after intervention study of a fusionless surgical technique to correct scoliosis secondary to spinal cord injury or myelodysplasia in children and adolescents. To determine the feasibility, safety, and utility of a fusionless treatment option for paralytic scoliosis. Once determined, these data could then be applied to develop the application of this operation for patients with other types of scoliosis, such as idiopathic. The optimal operative treatment for paralytic scoliosis remains to be determined. An ideal procedure would correct the deformity and stop the progression of scoliosis while maintaining mobility of the spine. This latter fact is important, especially for patients who rely heavily on use of trunk mobility for function. Fourteen patients with scoliosis secondary to spinal cord injury or myelodysplasia underwent a fusionless vertebral body wedge osteotomy procedure. Feasibility was analyzed by the ability to correct the scoliosis with the osteotomies and preserve mobility. Safety was reported by estimated blood loss, neurologic stability, and complications. Utility was reported by radiographic evidence of arrested curve progression and maintenance of spinal mobility. All 14 patients successfully underwent surgery to insert the wedge-rod system, with an average initial correction of 86% (range 66%-108%). The average estimated blood loss was 1050 cc (range 300-2000 cc). There were no major complications, and no changes in spasticity, bowel or bladder patterns, or motor/sensory levels. There was no case of nonunion at the osteotomy sites. At mean follow-up of 15 months (6-29 months), 10 patients had an improvement in their Cobb magnitude, 1 patient was within 5 degrees of their initial curve, 1 patient had a worse Cobb magnitude, and in 2 patients, the curve direction reversed but still measured less than the preoperative Cobb measurement. Spinal mobility was retained in all patients, as demonstrated on side-bending radiographs. The vertebral wedge osteotomy procedure appears to be a potential option for the treatment of paralytic scoliosis. The procedure was feasible and safely performed in these 14 patients, with spinal mobility maintained. There were no nonunions. The efficacy of the procedure is still not known, as is for which patients the procedure is indicated and timing of the operation. Long-term follow-up (to skeletal maturity) is needed. Only six of the patients are currently skeletally mature, and more numbers are needed to determine efficacy in this group.


Serum levels of SIRT3 and other inflammatory factors are associated with clinical outcomes and prognosis in severe community-acquired pneumonia in adults: A prospective study.
The aim of this study was to investigate clinical significance of SIRT3 in severe community-acquired pneumonia (CAP) patients.This prospective observational research enrolled a total of 114 severe CAP patients who went to our hospital during January 2018 to December 2019. Serum SIRT3 and IL-1β, IL-6, and tumor necrosis factor (TNF)-α levels were determined using the enzyme-linked immunosorbent assay (ELISA) method. Demographic data, including age, sex, and body mass index (BMI), as well as clinical symptoms, SOFA and SMART-COP scores were collected. The routine blood test was conducted for all patients and white blood cell (WBC) amount, as well as serum levels of C-reactive protein (CRP), D-Dimer, and procalcitonin (PCT).Among all patients, 55 cases died during the study period. The serum levels of CRP, PCT, IL-1β, and IL-6, as well as SOFA and SMART-COP scores were markedly higher in deceased patients than in the survival patients. The expression of SIRT3 was significantly decreased in severe CAP patients compared with the healthy, especially in the deceased patients. SIRT3 levels were negatively correlated with levels of CRP, PCT, IL-1β, and IL-6. Patients with SIRT3 low expression showed remarkably higher expression of CRP, PCT, IL-1β, and IL-6, as well as high SMART-COP scores, higher 1-month mortality rate, and shorter survival. Only SIRT3 and IL-1β were independent risk factors for 1-month mortality in severe CAP patients.Lower serum SIRT3 level predicts poor clinical outcomes and prognosis in severe CAP patients.


Copper-Mediated Intramolecular Interrupted CuAAC Selanylation.
We, herein, describe a copper-mediated domino CuAAC intramolecular selanylation for the synthesis of unprecedented fused benzo[4,5][1,3]selenazolo[3,2-c][1,2,3]triazoles from 1,2-bis(2-azidoaryl)diselenides and terminal alkynes under microwave irradiation. This is the seminal method for the synthesis of these fused heterocycles, and it proceeds under mild conditions, tolerates several functional groups, and can be carried out using environmentally benign solvents such as dimethyl carbonate. This transformation has been successfully extended to TMS-protected alkynes and to bioactive alkynes. A plausible reaction mechanism is proposed based on several control experiments and previous reports.


Principles of acute pain management.
Acute pain is a predominant feature of the perioperative experience for the majority of patients. This paper aims to describe the adverse effects of poorly controlled acute, postoperative pain and provides an overview of the organisational aspects involved in pain management in hospitals. Following this there will be an examination of the role information giving has in improving the patient's perioperative experience. Pharmacological and non-pharmacological interventions to prevent or reduce pain will also be described and because of its importance in setting the standard for logical prescribing in pain, the Analgesic Ladder, devised by the World Health Organisation (WHO 1986) will be given special consideration. Finally, the importance of pain assessment and re-assessment will be discussed.


Study on photoionization in a rubidium diode-pumped alkali laser gain medium with the optogalvanic method.
We use the optogalvanic method to calculate the concentration of rubidium ions produced by photoionization in a Rb diode-pumped alkali laser gain medium. With bias voltage added across the electrodes of a rubidium hollow cathode lamp, the measured optogalvanic current is 2.3×10(-7) A. Further study shows that the rubidium ion concentration is proportional to the pump intensity, and the drift velocity of rubidium ions is proportional to the bias voltage. When the photoionization process reaches dynamic equilibrium, the rubidium ion concentration will not increase with growing rubidium atom density. The calculated rubidium ion concentration is 1.5×10(5)-10(6) according to the experiment, and the ionization degree is less than 2.4×10(-7).


Sarcopenia and sarcopenic obesity.
The aging process is associated with progressive loss of muscle mass and strength, as well as decline in physical functioning. Although consensus diagnosis has not been reached, sarcopenia is increasingly defined by both loss of muscle mass and loss of muscle function or strength. The cause of sarcopenia is suggested as multifactorial, including hormonal changes, inflammatory pathway activation, fatty infiltration, poor nutrition, and decreased physical activity. Sarcopenia is often associated with visceral obesity. Sarcopenic obesity in the elderly impacts metabolic complications and represents a major public health challenge in a rapidly aging society. Further research about sarcopenia and sarcopenic obesity may be needed to confront the influence of aging society in Korea.


Alterations of bile acid and bumetanide uptake during culturing of rat hepatocytes.
Uptake by the multispecific bile acid transport system of [3H]taurocholate, [14C]cholate, and [3H]-bumetanide into primary cultures of rat hepatocytes was compared with their uptake into freshly isolated rat hepatocytes. The uptake maximum velocity (Vmax) of all compounds declined in primary culture, whereas the Michaelis constant (Km) values remained stable. Loss of uptake was not due to the reduction of driving forces as evaluated from the level of ATP and the activity of Na(+)-K(+)-ATPase. No alpha-fetoprotein was detectable in culture supernatants. Neither growth factors (glycylhistidyl-lysine, epidermal growth factor), peroxisome and cell proliferators (nafenopin, dimethyl sulfoxide), nor bile acids prevented the loss of transport in hepatocyte culture. However, addition of dibutyryl adenosine 3'5'-cyclic monophosphate protracted the transport activity significantly. When cultured rat hepatocytes with reduced transport were detached by trypsin, cells rounded up and showed the same uptake capacity for bumetanide, cholate, and taurocholate as seen in freshly isolated hepatocytes. "Cryptic" transport activity in the lower basolateral membrane facing the support was found using an incubation chamber for cultured hepatocytes, which allowed us to distinguish simultaneously between uptake via the upper and lower basolateral membrane of the cultured cells.


Cardiopulmonary and arterial baroreflex responses to acute volume expansion during fetal and postnatal development.
Recent studies demonstrated that renal denervation had no effect on the natriuretic response to volume expansion (VE) in fetal sheep, suggesting that the sensitivity of the cardiopulmonary reflex in response to VE is impaired in the fetus. To test this hypothesis, we investigated the renal sympathetic nerve activity (RSNA) and heart rate (HR) responses to 20 and 50% intravascular VE in fetal (130-135 days gestation; term 145 days) (n = 7), newborn (n = 8), and 6- to 8-wk-old sheep (n = 9). Despite similar increases in right atrial pressure (RAP) in the three groups, 20% VE had no significant effect on RSNA and HR in fetal sheep but significantly decreased RSNA in newborn (-22.8 +/- 7.3%) and 6- to 8-wk-old sheep (-32.1 +/- 11.7%). Bradycardic responses to VE were also observed in both newborn (from 237 +/- 6 to 200 +/- 12 beats/min) and 6- to 8-wk-old sheep (from 170 +/- 9 to 140 +/- 9 beats/min). A 50% VE had no significant effect on fetal RSNA and HR, whereas it increased RAP by 6.8 +/- 0.9 mmHg. In addition, we tested the hypothesis that interactions between cardiopulmonary and arterial baroreflexes in response to VE change during development. We found that 20 and 50% VE shifted the RSNA and HR arterial baroreflex response curves to the right in the fetus but had no significant effects on the gain of the arterial baroreflex curves in either fetal, newborn, or 6- to 8-wk-old sheep.(ABSTRACT TRUNCATED AT 250 WORDS)


Bacteriology of chronic sinusitis: the bulla ethmoidalis content.
To study the microbiology of the bulla ethmoidalis of patients who suffered from chronic sinusitis. A prospective study performed at the Saint-Luc universitary clinic (University of Louvain) from June 1999 to December 2000. Seventy seven patients underwent functional endoscopic endonasal surgery during this period for chronic sinusitis refractory to standard medical treatment. All the patients presented symptoms for more than 3 months. After Povidone-Iodine disinfection, samples were taken into the bulla ethmoidalis after its opening with an endoscopic endonasal control. Samples were transferred to the microbiology laboratory using a thioglycolate medium for aerobic and anaerobic cultures. One hundred forty eight samples were studied. Culture rate was 73.6%. Thirty nine samples remained sterile. In the 109 culture positive specimens, 135 bacterial isolates were recovered. The main results are: Staphylococcus coagulase negative: 31, Staphylococcus aureus: 22, Streptococcus sp: 20, other Gram positive Cocci: 5, Haemophilus influenzae: 4, non-fermentative Gram negative bacilli: 6, Enterobacteriaceae: 45, anaerobic bacteria: 2. Enterobacteriaceae or enteric gram negative bacilli were frequently encountered in the bulla ethmoidalis of patients suffering from chronic sinusitis. This report suggests that endoscopically guided culture obtained from the ethmoid sinus may accurate our understanding of the microbiology of chronic ethmoidal sinusitis and underline the importance of Enterobacteriaceae in this disease.


Cellular and molecular choreography of neutrophil recruitment to sites of sterile inflammation.
Liberation of damage-associated molecular patterns (DAMPs) following tissue injury and necrotic cell death leads to the induction of sterile inflammation. A hallmark of acute inflammation is the recruitment of neutrophils to injured tissues. This review focuses on the journey of neutrophils to sites of sterile inflammation and the cellular and molecular mechanisms that choreograph this complex voyage. We review the pathway of leukocyte recruitment, with emphasis on recent additions to our understanding of intravascular neutrophil migration. The contributions of various tissue-resident sentinel cell populations to the detection of danger signals (DAMPs) and coordination of neutrophil recruitment and migration are discussed. In addition, we highlight recent data on the control of neutrophil chemotaxis towards sites of sterile inflammation, including new insight into the temporal and spatial regulation of chemoattractant guidance signals that direct cell migration. Given that inappropriate neutrophilic inflammation is a cornerstone in the pathogenesis of many diseases, a complete understanding of the choreography of neutrophil recruitment to sites of sterile inflammation may uncover new avenues for therapeutic interventions to treat inflammatory pathologies.


ANESTHESIA FOR ROBOT-ASSISTED RADICAL PROSTATECTOMY - A CHALLENGE FOR ANAESTHESIOLOGIST.
Mininimally invasive surgery has become one of the most popular ones over the last few decades due to many benefits. The advantages are minimal surgical incision, reduced blood loss, reduced postoperative pain, faster postoperative recovery, shorter hospital stay, lower morbidity and better outcomes compared to open surgery. The most common robotic procedures in urology are radical prostatectomies. In UHC Zagreb, since November 2019 until now, there have been more than 180 robotic assisted radical prostatectomies (RALP) using Senhance robotic system performed. As a procedure with many possible complications, it represents a challenge for anaesthesiologist. Some of the problems the anaesthesiologists have to face are related to limited patient access, possible difficulties connected with positioning, pneumoperitoneum, subcutaneous emphysema, possible airway oedema. Pneumoperitoneum has impact on almost every system: cardiovascular, renal, respiratory, gastrointestinal and other. Detailed understanding of physiological changes of RALP, with intraoperative impact on nearly every body system is ultimate. Careful preoperative evaluation and intraoperative conduction minimize the risk of complications, and help patients to reach full recovery in a very short time. Excellent outcomes are the result of individualized approach to the patient and good communication between team members.


Current status of bioresorbable scaffolds in the treatment of coronary artery disease.
State-of-the-art drug-eluting metal stents are the gold standard for interventional treatment of coronary artery disease. Although they overcome some disadvantages and limitations of plain balloon angioplasty and bare-metal stents, some limitations apply, most notably a chronic local inflammatory reaction due to permanent implantation of a foreign body, restriction of vascular vasomotion due to a metal cage, and the risk of late and very late stent thrombosis. The development of biodegradable scaffolds is a new approach that attempts to circumvent these drawbacks. These devices provide short-term scaffolding of the vessel and then dissolve, which should theoretically circumvent the side effects of metal drug-eluting stents. Various types of these bioresorbable scaffolds are currently under clinical evaluation. This review discusses different concepts of bioresorbable scaffolds with respect to material, design, and drug elution and presents the most recent evidence.


[Social health insurance in China: principal reforms and inequalities].
This article analyzes the social health insurance system in China, its reforms and the principal social inequalities uncovered. Based in the work of a number of authors of reference, it is possible to observe that rural and urban reforms follow the same pattern: large systems that were gradually reduced and then again expanded relatively quickly. Improvements notwithstanding, some of China's historical problems persist, especially the rural-urban gap and regional disparities. The lack of integration of workers that migrate from the country to the city is reproduced in the current Chinese public health system, constituting one of the primary challenges to be faced at present.


How to improve the incorporation of massive allografts?
The incorporation of a bone graft is the result of creeping and substitutional activities that remove the original grafted bone and replace it by newly formed bone from the host cells. However, this intricate process is very limited in time and space. A bone allograft is poorly remodeled and is almost non viable even after several years of implantation. This lack of vitality accounts for the high rate of complications such as non union and fracture. One way to minimize the allograft complications is to improve its incorporation. The process of incorporation in animals and human beings is reviewed as well as the various avenues for a biologic improvement either through modulation on the host: the immune response, the inhibition of bone resorption, the use of bone morphogenetic proteins, the autogenous cell augmentation or through processing the bone allograft: bisphosphonate adsorption or bone perforations. In 2002, biologic enhancement of the incorporation is still in its infancy but will be in a near future a reality through influence on both the host and the allograft.


Can Exchange Transfusion Normalize Serum Levels of Copper, Zinc, and Magnesium in Severe Neonatal Hyperbilirubinemia?
Neonatal hyperbilirubinemia is a frequently encountered problem. Erythrocytes, especially reticulocytes are rich in copper (Cu) and magnesium (Mg) so its serum levels increase after hemolysis. Zinc (Zn) plays an important role in synthesis of some enzymes included in the bilirubin metabolism and may cause hemolysis. Exchange transfusion is the main treatment for severe neonatal hyperbilirubinemia but can exchange transfusion affect the previous trace elements. We measured Cu, Zn, and Mg serum levels in full-term neonates admitted to neonatal intensive care unit of Minia University hospital with severe indirect hyperbilirubinemia before and after exchange transfusion. There were significant higher serum Cu and Mg and lower Zn serum levels in neonates with hyperbilirubinemia than controls and their levels were significantly normalized after exchange transfusion. Significant positive correlations between the total bilirubin levels and hemoglobin, Cu and Mg serum levels and significant negative correlations with Zn levels were present. There were no significant correlations between maternal and neonatal serum levels of any of them. Exchange transfusion can normalize the significant higher Cu and Mg and lower Zn serum levels in neonates with severe indirect hyperbilirubinemia which were not related to their maternal serum levels.


A comparative analysis of the HUGOTM robot-assisted surgery system and the Da Vinci® Xi surgical system for robot-assisted sacrocolpopexy for pelvic organ prolapse treatment.
We aimed to compare the outcomes of Robot-assisted sacrocolpopexy (RASC) performed using the novel HUGOTM Robot-Assisted Surgery (RAS) System with the Da Vinci® Xi surgical system. Data from 38 women undergoing RASC for a ≥ 2-grade pelvic organ prolapse were collected (2021-2023). Overall, 23 (60.5%) and 15 (39.5%) procedures were performed using the DaVinci® Xi and the HUGOTM RAS system, respectively. The median total operative time was 123 (IQR:106.5-140.5) minutes for the DaVinci® Xi versus 120 (IQR:120-146) minutes for the HUGOTM RAS cases (p = 0.5). No conversion to open/laparoscopic surgery, perioperative complications, or system failures occurred. No differences were recorded according to day of catheter removal and length of stay. This study represents the first worldwide comparison of RASC executed using the HUGOTM RAS versus the Da Vinci® Xi System. Our data suggest that RASC might be performed with both robotic platforms with similar perioperative outcomes.


Metabolic zonation of liver parenchyma.
Periportal and perivenous hepatocytes differ in their content of enzymes and subcellular structures and thus in their metabolic capacities. Therefore, the model of metabolic zonation proposes that: (1) periportal hepatocytes catalyze predominantly oxidative energy metabolism with beta-oxidation and amino acid catabolism as well as ureagenesis for glycogen synthesis and glucose release, bile formation with cholesterol synthesis and protective metabolism; and (2) perivenous hepatocytes mediate preferentially glucose uptake for glycogen synthesis, glycolysis, and liponeogenesis as well as ketogenesis, glutamine formation, and xenobiotic metabolism. Periportal and perivenous hepatocytes are under the control of a different input of humoral and nervous signals, because concentration gradients of oxygen, substrates, and hormones are established during passage of blood through the liver and because gradients of nerve densities seem to exist. In periportal and perivenous hepatocytes gene expression can be different due to the zonal gradients in oxygen and hormone concentrations as well as nerve densities. The functional specialization of periportal and perivenous hepatocytes has been demonstrated especially well for carbohydrate, amino acid and ammonia, and xenobiotic metabolism as well as for bile formation by different techniques: Calculation of metabolic rates based on the zonal distributions of enzymes and metabolites, measurements of rates in periportal-like and perivenous-like hepatocytes in cell culture and in hepatocyte populations enriched in periportal and perivenous cells as well as in perfused livers during orthograde and retrograde flow using standard methods and noninvasive techniques with surface microlight guides and miniature oxygen electrodes.


Dysmorphogenic effects of some fungicides derived from the imidazole on rat embryos cultured in vitro.
Like triazole-derivatives, imidazole-derivatives exert their antifungal and toxicological properties by inhibiting P450 enzymes (Cyps). At the embryonic level, Cyp enzymes are involved also in the catabolism of the retinoic acid. Specific effects of triazole-derivatives have been reported on developing rodent embryos, and were correlated to an imbalance of the retinoid homeostasis. The aim of this work was to investigate if imidazole-derivatives are able to induce specific malformations similar to those observed after triazole-derivative exposure. Post implantation rat embryos were exposed in vitro to 1,000 microM Imidazole and to 5-100 microM of the imidazole-derivatives Ketoconazole and Enilconazole. After 48 h in culture, the embryos exposed to the imidazole-derivatives showed specific malformations, quite similar to those observed after triazole-derivative exposure. The common dysmorphogenic effects of the azole-derivatives of the two classes could be due to the inhibition of retinoid catabolism. From this point of view, the contemporaneous exposure to these substances or their therapeutic use could be considered as potentially dangerous for human conceptuses.


gamma1 heavy chain transgenes are responsive to IFN-gamma repression and CD40 ligation.
The cis-acting elements that regulate production of germ-line transcripts from Ig heavy chain genes and subsequent switch recombination to those genes are poorly defined. We reported that a 17-kb transgene that includes Igamma1, Sgamma1, and Cgamma1 is regulated for germ-line transcription like the endogenous gamma1 gene. Transcripts from such transgenes are expressed only in B cells treated with both LPS and IL-4, and not in B cells treated with LPS alone or in thymocytes or nonlymphoid tissues. We have now found that transcripts from these transgenes are induced by treatment of transgenic B cells by IL-4 alone. As reported by others, IFN-gamma acts to inhibit the IL-4-mediated induction of germ-line transcripts of the endogenous gamma1 gene. We have found that LPS-plus IL-4-induced germ-line transcription of gamma1 transgenes is likewise inhibited by treatment of B cells with IFN-gamma, so the gamma1 gene must include the cis-acting element(s) that confers this inhibition. It is also known that CD40 ligation induces a modest amount of germ-line transcripts from the endogenous gamma1 gene and synergizes with IL-4 to induce large amounts of germ-line transcripts. The gamma1 transgenes are likewise induced by CD40 ligation, suggesting that the response element(s) for CD40 ligation can be found in the gamma1 gene. The promoter region for the germ-line transcripts and the I exon are likely to include some of these cis-acting elements. A series of transgenic mice with the promoter/Igamma1 region conferred low level, lymphoid-specific, RNA expression to a reporter gene, including significant expression in thymocytes. However, the promoter/Igamma1 transgenes were not regulated like the endogenous gamma1 gene, in that transcription in splenic B cells was not increased by LPS plus IL-4.


New scaffolds for supramolecular chemistry: upper-rim fully tethered 5-methyleneureido-5'-methyl-2,2'-bipyridyl cyclodextrins.
Seven upper-rim fully tethered cyclodextrins (URFT-CDs) have been synthesised in a good average coupling yield using the one-step "phosphine imide" approach and their metal complexation behaviour with lanthanides and transition metals was explored. We observed that the A-TE-E light conversion process (antennae effect) occurs in the URFT-CD lanthanide complexes. A molecular redox switch based on the corresponding iron complexes is also reported. A reversible intramolecular translocation of the FeII and FeIII ions, between two distinct binding cavities has been monitored spectroscopically and achieved by chemical triggering. Finally, a negative allosteric control of ion recognition through the formation of a CD pseudocryptand is discussed.


How to see the trees for the forest: introduction to a special issue on causation and disease.
This paper summarizes the results from the first European Advanced Seminar in the Philosophy of the Life Sciences, which was held at the Brocher Foundation in Hermance (Switzerland) 6-10 September 2011. The Advanced Seminar brought together philosophers of the life sciences to discuss the topic of "Causation and Disease." The search for causes of disease in the biomedical sciences, we argue on the basis of the contributions to this conference, has not resulted in a simplification and unification of biomedical knowledge, as once hoped for by philosophers of science, but rather in its "complexification."


[Molecular cloning and analysis of the 3' terminal sequence of duck hepatitis virus genome].
The authentic 3' terminal sequences of genomes of duck hepatitis virus (DHV) serotype I strain C80 and serotype I variant strain E63 were obtained by using 3' RACE and RT-PCR techniques. The analysis showed that 3' terminal sequences in the genomes of the two DHV strains include the 3D region of 1359 nucleotides (nt), the terminator codon TGA, and 3'untranslated region (UTR) of 314 nt. The poly (A) tails of C80 and E63 are 18 nt and 19 nt in length respectively. The deduced 3D proteins of 453 amino acids of both DHV strains contain the motifs KDELR, DxxxxD, GxxCSGxxxTxxxNS, YGDD, and FLKR characteristic for RNA polymerase of picornaviruses, which confirms DHV serotype I to be a member of Picornaviridae . The amino acid sequence identities among 3D protein of the two DHV strains with representatives of nine other picornavirus genera range from 16% to 37%, which are within the value ranges (18%-60%) of 3D amino acid sequence identities obtained from inter-genera comparisons. In addition, DHV serotype I possesses the longest 3'UTR in the family Picornaviridae. Phylogenetic analysis of 3D proteins indicated that DHV serotype I may belong to a separated genus of the family Picornaviridae.


A proteomic primer for the clinician.
Proteomics is a rapidly developing field and it opens new horizons in many research areas of life sciences. In the field of medicine, proteomics promises to accelerate the discovery of new drug targets and protein disease markers useful for in vitro diagnosis. In this article, we review the current proteomics technologies for biomarker discovery and validation, which include two-dimensional gel electrophoresis, one- and two-dimensional liquid chromatography, and proteomic microarrays. We will also review proteomic strategies for protein-protein interactions and identification of post-translational modifications. Selection of the more effective technology or combination of technologies is required to maximize the interpretation and utility of the data.


Caught in the act: structural dynamics of replication origin activation and fork progression.
This review discusses recent advances in single-particle cryo-EM and single-molecule approaches used to visualise eukaryotic DNA replication reactions reconstituted in vitro. We comment on the new challenges facing structural biologists, as they turn to describing the dynamic cascade of events that lead to replication origin activation and fork progression.


A prospective, multicenter, three-cohort study evaluating contrast-induced acute kidney injury (CI-AKI) in patients with cirrhosis.
Nephrotoxicity of intravenous iodinated contrast media (ICM) in cirrhosis is still a debated issue, due to scarce, low-quality and conflicting evidence. This study aims to evaluate the incidence and predisposing factors of acute kidney injury (AKI) in patients with cirrhosis undergoing contrast-enhanced computed tomography (CECT). We performed a prospective, multicenter, cohort study including 444 inpatients, 148 with cirrhosis (cohort 1) and 163 without cirrhosis (cohort 3) undergoing CECT and 133 with cirrhosis (cohort 2) unexposed to ICM. Kidney function parameters were assessed at T0, 48-72 h (T1), 5 and 7 days after CECT/enrollment. Urinary neutrophil gelatinase-associated lipocalin (U-NGAL) was measured in 50 consecutive patients from cohort 1 and 50 from cohort 2 as an early biomarker of tubular damage. AKI incidence was not significantly increased in patients with cirrhosis undergoing CECT (4.8%, 1.5%, 2.5% in cohorts 1, 2, 3 respectively, p = n.s.). Most AKI cases were mild and transient. The presence of concomitant infections was the only independent predictive factor of contrast-induced AKI (odds ratio 22.18; 95% CI 2.87-171.22; p = 0.003). No significant modifications of U-NGAL between T0 and T1 were detected, neither in cohort 1 nor in cohort 2 (median ΔU-NGAL: +0.2 [-7.6 to +5.5] ng/ml, +0.0 [-6.8 to +9.5] ng/ml, respectively [p = 0.682]). AKI risk after CECT in cirrhosis is low and not significantly different from that of the general population or of the cirrhotic population unexposed to ICM. It mostly consists of mild and rapidly resolving episodes of renal dysfunction and it is not associated with tubular kidney injury. Patients with ongoing infections appear to be the only ones at higher risk of AKI. Nephrotoxicity due to intravenous iodinated contrast media (ICM) in patients with cirrhosis is still a debated issue, as the available evidence is limited and based on very heterogeneous studies, often conducted on small and retrospective cohorts. In this prospective three-cohort study we found that intravenous administration of ICM was associated with a low risk of AKI, similar to that of the general population and to that of patients with cirrhosis unexposed to ICM. Patients with ongoing infections were the only ones to have a significantly increased risk of contrast-induced AKI. Therefore, the actual recommendations of performing contrast imaging studies cautiously in cirrhosis do not seem to be reasonable anymore, with the exception of infected patients, who have a significantly higher risk of contrast-induced AKI.


Bilateral Choroidal Detachment Induced by Unilateral Application of a Fixed Combination of Topical Timolol Maleate and Brinzolamide.
We describe a 66-year-old man who developed bilateral choroidal detachment that was induced by unilateral topical administration of a fixed combination of 1% brinzolamide and 0.5% timolol maleate the day after an uneventful phacoemulsification surgery and intraocular lens implantation involving his right eye. We believe that the reaction was an idiosyncratic reaction, most likely against brinzolamide. The condition improved rapidly after the cessation of the fixed combination of brinzolamide and timolol maleate and treatment with 1% topical prednisolone acetate every hour and 1% cyclopentolate twice a day bilaterally. Although there are several similar cases involving choroidal detachment after oral acetazolamide and topical dorzolamide treatment mentioned in the literature, the present case is the first case report involving bilateral choroidal detachment after topical treatment with brinzolamide.


Nickel stabilization efficiency of aluminate and ferrite spinels and their leaching behavior.
Stabilization efficiencies of spinel-based construction ceramics incorporating simulated nickel-laden waste sludge were evaluated and the leaching behavior of products investigated. To simulate the process of immobilization, nickel oxide was mixed alternatively with gamma-alumina, kaolinite, and hematite. These tailoring precursors are commonly used to prepare construction ceramics in the building industry. After sintering from 600 to 1480 degrees C at 3 h, the nickel aluminate spinel (NiAl204) and the nickel ferrite spinel (NiFe204) crystallized with the ferrite spinel formation commencing about 200-300 degrees C lower than for the aluminate spinel. All the precursors showed high nickel incorporation efficiencies when sintered at temperatures greater than 1250 degrees C. Prolonged leach tests (up to 26 days) of product phases were carried out using a pH 2.9 acetic acid solution, and the spinel products were invariably superior to nickel oxide for immobilization over longer leaching periods. The leaching behavior of NiAl2O4 was consistent with congruent dissolution without significant reprecipitation, but for NiFe2O4, ferric hydroxide precipitation was evident. The major leaching reaction of sintered kaolinite-based products was the dissolution of cristobalite rather than NiAl2O4. This study demonstrated the feasibility of transforming nickel-laden sludge into spinel phases with the use of readily available and inexpensive ceramic raw materials, and the successful reduction of metal mobility under acidic environments.


miR-186-ANXA9 signaling inhibits tumorigenesis in breast cancer.
Breast cancer (BC) ranks as the highest incidence among cancer types in women all over the world. MicroRNAs (miRNAs) are a class of short endogenous non-coding RNA in cells mostly functioning to silence the target mRNAs. In the current study, a miRNA screening analysis identified miR-186-5p to be downregulated in human breast cancer tumors. Functional studies in vitro demonstrated that overexpression of miR-186-5p inhibited cellular proliferation and induced cell apoptosis in multiple breast cancer cell lines including MDA-MB-231, MCF-7, and BT549 cells. Transplantation of the miR-186-5p-overexpressing MDA-MB-231 cells into nude mice significantly inhibited mammary tumor growth in vivo. Sequence blast analysis predicted annexin A9 (ANXA9) as a target gene of miR-186-5p, which was validated by luciferase reporter assay, QRT-PCR analysis, and western blot. Additional gene expression analysis of clinical tumor samples indicated a negative correlation between miR-186-5p and ANXA9 in human breast cancer. Knockdown of ANXA9 mimicked the phenotype of miR-186-5p overexpression. Reintroduction of ANXA9 back rescued the miR-186-5p-induced cell apoptosis. In addition, miR-186-5p decreased the expression of Bcl-2 and increased the expression of p53, suggesting a mechanism regulating miR-186-5p-induced cellular apoptosis. In summary, our study is the first to demonstrate miR-186-5p-ANXA9 signaling in suppressing human breast cancer. It provided a potential therapeutic target in breast cancer.


Pathophysiological and clinical aspects of the circadian rhythm of arterial stiffness in diabetes mellitus: A minireview.
Several cross-sectional trials have revealed increased arterial stiffness connected with the cardiac autonomic neuropathy in types 2 and 1 diabetic patients. The pathophysiological relationship between arterial stiffness and autonomic dysfunction in diabetes mellitus is still underinvestigated and the question whether the presence of cardiac autonomic neuropathy leads to arterial stiffening or increased arterial stiffness induced autonomic nervous system impairment is still open. Both arterial stiffness and dysfunction of the autonomic nervous system have common pathogenetic pathways, counting state of the chronic hyperinsulinemia and hyperglycemia, increased formation of advanced glycation end products, activation of protein kinase C, development of endothelial dysfunction, and chronic low-grade inflammation. Baroreceptor dysfunction is thought to be one of the possible reasons for the arterial wall stiffening development and progression. On the contrary, violated autonomic nervous system function can affect the vascular tone and by this way alter the large arteries walls elastic properties. Another possible mechanism of attachment and/or development of arterial stiffness is the increased heart rate and autonomic dysfunction corresponding progression. This minireview analyzes the current state of the relationship between the diabetes mellitus and the arterial stiffness. Particular attention is paid to the analysis, interpretation, and application of the results obtained in patients with type 2 diabetes mellitus and diabetic cardiac autonomic neuropathy.


[The corpus luteum].
The primary function of the corpus luteum is secretion of progesterone for maintenance of pregnancy. The development and function of the corpus luteum from residual follicular granulosa and theca cells after ovulation is induced by the midcyclic peak of LH secretion followed by further pulsatile LH release. Due to this stimulation the follicular granulosa and theca cells are converted to large and small luteinized cells with high proliferation rate. During this process Vascular-Endothelial-Growth Factor (VEGF) plays a major role as a potent stimulator of neo-angiogenesis. Formation of new blood vessels is essential to ensure supply of LDL-Cholesterol as substrate for steroidogenesis. If pregnancy does not occurs, the corpus luteum must regress to initiate another cycle. Luteal regression seems to be initiated by PGF2 alpha which is secreted from the uterus. PGF2 alpha, reduces luteal blood flow and progesterone synthesis. Furthermore it is a potent inducer of apoptosis. If pregnancy occurs, sustained secretion of progesterone and other substances like estradiol and relaxin are required to provide an appropriate uterine environment for maintenance of pregnancy. In that case the corpus luteum is further stimulated by hCG secreted by the blastocyst and the trophoblast-cells until 8/9 weeks of gestational age, when synthesis and secretion of steroids is taken over by the placenta.


A rapid LC-MS/MS method for quantification of CSUOH0901, a novel antitumor agent, in rat plasma.
CSUOH0901, a novel anticancer derivative of nimesulide, exhibits very promising anticancer activities in various cancer cell lines. In order to support further pharmacological and toxicological studies of this promising anticancer drug candidate, an LC-MS/MS method was developed and validated in accordance with the US Food and Drug Administration guidelines. The drug molecules were extracted from plasma samples by protein precipitation and then analyzed with LC-ESI-MS/MS. An excellent analyte separation was achieved using a phenomenex C18 column with a mobile phase of 90% methanol and 5 m m of ammonium formate. The validated linear dynamic range was between 0.5 and 100 ng/mL and the achieved correlation coefficient (r(2)) was >0.9996. The results of inter- and intra-day precision and accuracy were satisfactory, that is, <12% for accuracy and within ±5% for precision at a low and high quality control concentrations, respectively. In addition, the analyte and internal standard (JCC76) were found to be stable under the storage conditions at -20°C for about 2 months. Hence, the acquired results proved that the LC-ESI-MS/MS method developed is precise, accurate and selective for the quantification of CSUOH0901 in plasma, and can be used for pharmacokinetic studies.


Mediterranean diet, cardiovascular disease and mortality in diabetes: A systematic review and meta-analysis of prospective cohort studies and randomized clinical trials.
To update the clinical practice guidelines for nutrition therapy of the European Association for the Study of Diabetes, we conducted a systematic review and meta-analysis of prospective cohort studies and randomized clinical trials (RCTs) to evaluate the effect of the Mediterranean diet (MedDiet) on the prevention of cardiovascular disease (CVD) incidence and mortality. We searched Medline, EMBASE (through April 20, 2018) and Cochrane (through May 7, 2018) databases. Pooled relative risks (RRs) and 95% confidence interval (CI) were calculated by the generic inverse variance method. A total of 41 reports (3 RCTs and 38 cohorts) were included. Meta-analyses of RCTs revealed a beneficial effect of the MedDiet on total CVD incidence (RR: 0.62; 95% CI: 0.50, 0.78) and total myocardial infarction (MI) incidence (RR: 0.65; 95% CI: 0.49, 0.88). Meta-analyses of prospective cohort studies, which compared the highest versus lowest categories of MedDiet adherence, revealed an inverse association with total CVD mortality (RR: 0.79; 95% CI: 0.77, 0.82), coronary heart disease (CHD) incidence (RR: 0.73; 95% CI: 0.62, 0.86), CHD mortality (RR: 0.83; 95% CI: 0.75, 0.92), stroke incidence (RR: 0.80; 95% CI: 0.71, 0.90), stroke mortality (RR: 0.87; 95% CI: 0.80, 0.96) and MI incidence (RR: 0.73; 95% CI: 0.61, 0.88). The present study suggests that MedDiet has a beneficial role on CVD prevention in populations inclusive of individuals with diabetes.


A novel cis-peptide bond motif inducing beta-turn type VI. The synthesis of enkephalin analogues modified with 4-aminopyroglutamic acid.
A new pathway leading to a mixture of four isomers of 4-aminopyroglutamic acid is described. Michael type addition of Z-deltaAla-OMe to enolates prepared from acylaminomalonates, followed by hydrolysis and decarboxylation give protected 4-aminopyroglutamic acid with the cis:trans ratio approximately 3:2. This mixture was incorporated into Leu-enkephalin (position 2-3). After separation of peptides it appeared that all analogues were essentially inactive in guinea pig ileum and mouse vas deferens bioassays.


Thrombin generation by hemolysis.
Hemolysis is the fragmentation of erythrocytes into microparticles (Hb-MP). Clinical hemolysis can result in a severe procoagulant state. The influence of Hb-MP on thrombin generation was quantified. Unfrozen citrated normal plasma (five donors) was supplemented with 0 or 1 g/l Hb-MP obtained through erythrocyte destruction by hypotonic lysis, freezing/thawing, or blood oxidation with 1 or 2 mmol/l chloramine-T. Pooled normal plasma was supplemented with 0-10 g/l Hb-MP and with 0-1 IU/ml low-molecular-weight heparin (dalteparin). Samples (50 microl) were tested in the recalcified coagulation activity assay. At 10 min coagulation reaction time the hypotonic lysis of erythrocytes appears to be the most procoagulant condition, followed by twice freezing/thawing, three times freezing/thawing, and once freezing/thawing. Oxidation of whole blood with 1 or 2 mmol/l chloramine-T decreases thrombin generation by about 20 or 50%, respectively. The thrombin generation in 1 mmol/l chloramine-T or 2 mmol/l oxidized plasma decreases by about 70 or 85%, respectively. The 50% inhibitory concentrations of low-molecular-weight heparin against recalcified thrombin generation are 0.01, 0.025, or 0.035 IU/ml for plasma supplemented with 0, 0.1, or 1 g/l Hb-MP, respectively. The recalcified coagulation activity assay allows one to quantify thrombin generation in critical hemolytic samples. It is suggested to find the appropriate pharmacologic dose of low-molecular-weight heparin.


[Effect of acyclovir and steroid in a young immunocompetent male with herpes zoster myelitis].
Herpes zoster myelitis is a rare condition, usually seen in aged and immunocompromised patients. Due to atypical presen-tations it can be hard to diagnose. Intraspinal lesions on magnetic resonance imaging (MRI) support the diagnosis. We present a 39-year-old otherwise healthy male with symptoms of viral meningitis and rapidly progressing symptoms of myelitis. Lumbar puncture showed increased levels of monocytes and varicella zoster virus DNA. Despite a negative MRI, based on a few previous case reports and because of lack of progress on antiviral treatment, treatment with steroids was established early, recovering the patient dramatically. This supports that a combination of antiviral treatment and steroids may be a more efficient treatment of zoster myelitis and reminds us that the diagnosis cannot be excluded by a negative MRI.


Immune stimulatory effects of neutrophil extracellular traps in granulomatosis with polyangiitis.
The aim of this study was to analyse the role of netting neutrophils in the pathogenesis of granulomatosis with polyangiitis (GPA), especially their interplay with peripheral blood mononuclear cells (PBMCs). The amount of cell-free DNA (cfDNA) was determined in sera from GPA patients (pairs active/inactive state of disease, n=18) and from healthy controls (HCs, n=10). Furthermore, we performed in vitro incubation experiments using PBMCs and NETs from patients and HCs for accessing the effect of NETs on PBMC behaviour. We determined proliferation of T- and B-cells (CSFE assay), B-cell maturation (CD38 staining and flow cytometry), production of IgG (ELISpot, ELISA), and secretion of the cytokines IFN-γ, IL-4, IL-10, IL-17A (ELISA). We detected a significant increase in serum cfDNA levels of GPA patients compared to HCs. The concentration of cfDNA was associated with disease activity. NETs of patients and HCs induced proliferation of CD4+ T- cells and CD19+ B-cells and maturation of B-cells. Furthermore, we detected an increase in IL-17A secretion after stimulating PBMCs with NETs. A significant difference between PBMCs from GPA patients and HCs was not detectable. NETs activate PBMCs of HCs and GPA patients. Our findings give supportive evidence that NETosis plays a role in the pathogenesis of GPA.


Pre- and Postnatal Effects of Corticosterone on Fitness-Related Traits and the Timing of Endogenous Corticosterone Production in a Songbird.
Maternally derived corticosterone in the egg and corticosterone produced endogenously by altricial nestling birds play essential roles during development. Although persistently high corticosterone levels can be harmful, moderately elevated levels above baseline can lead to reallocation of resources between growth and maintenance to ensure immediate survival or to enhance the development of fitness-related traits. We tested two hypotheses concerning the fitness consequences of elevated corticosterone during prenatal and postnatal development in altricial house wrens: (1) elevated corticosterone shifts resources away from growth and immune function and (2) elevated corticosterone serves as a signal to allocate resources to fitness-related traits. We also explored the development of the stress response, hypothesizing that early-stage nestlings have little endogenously produced corticosterone, but that their baseline and stress-induced corticosterone levels increase with age. Nestlings hatching from corticosterone-injected eggs were lighter at hatching, but through compensatory growth, ended up heavier than controls near the time of fledging, an important, fitness-related trait. Nestlings that hatched from corticosterone-injected eggs and those given oral doses of corticosterone did not differ from controls in three other fitness-related traits: immunoresponsiveness, size, or haematocrit. Early- and late-stage nestlings had similar baseline corticosterone levels, and all nestlings increased their plasma corticosterone levels in response to a capture-and-restraint protocol, with older nestlings mounting a stronger stress-induced response than younger nestlings. These results suggest that prenatal exposure to corticosterone is important in shaping offspring phenotype and are consistent with the hypothesis that maternally derived corticosterone in the egg can have long-term, fitness-related effects on offspring phenotype.


Retardation of calcium oxalate formation by polyacidic peptides.
The polycarboxylic amino acids were examined as possible retarding agents in the precipitation of calcium oxalate. The effect was observed by continuous determination of the concentration of calcium ions in a mimic urine solution. Both polyglutamic and polyaspartic acids, when used in 5 to 100 ppm concentrations, significantly retarded calcium oxalate precipitation.


A Randomized Controlled Trial of the Rochester Forensic Assertive Community Treatment Model.
Forensic assertive community treatment (FACT) is an adaptation of the assertive community treatment model and is designed to serve justice-involved adults with serious mental illness. This study compared the effectiveness of a standardized FACT model and enhanced treatment as usual in reducing jail and hospital use and in promoting engagement in outpatient mental health services. Seventy adults with psychotic disorders who were arrested for misdemeanor crimes and who were eligible for conditional discharge were recruited from the Monroe County, New York, court system. Participants were randomly assigned to receive either FACT (N=35) or enhanced treatment as usual (N=35) for one year. Criminal justice and mental health service utilization outcomes were measured by using state and county databases. Forty-nine participants (70%) completed the full one-year intervention period. Nineteen (27%) were removed early by judicial order, one was removed by county health authorities, and one died of a medical illness. Intent-to-treat analysis for all 70 participants showed that those receiving the FACT intervention had fewer mean±SD convictions (.4±.7 versus .9±1.3, p=.023), fewer mean days in jail (21.5±25.9 versus 43.5±59.2, p=.025), fewer mean days in the hospital (4.4±15.1 versus 23.8±64.2, p=.025), and more mean days in outpatient mental health treatment (305.5±92.1 versus 169.4±139.6, p<.001) compared with participants who received treatment as usual. The Rochester FACT model was associated with fewer convictions for new crimes, less time in jail and hospitals, and more time in outpatient treatment among justice-involved adults with psychotic disorders compared with treatment as usual.


Case report: the role of OCT in examination of a patient with topiramate-induced acute angle closure, acute myopia and macular striae.
This work reports on a clinical case of a female who presented with headache, bilateral eye pain and vision loss. Intraocular pressures were 40 mm Hg in the right eye and 45 mm Hg in the left eye. Optical Coherence Tomography examination shows the iridocorneal angle was collapsed and macular striae were also observed. The patient had been on topiramate due to migraines 7 days before presentation. Diagnosis for topiramate-induced acute angle closure was made in both eyes. The patient showed improvement in symptoms a few days after treatment initiation and images confirmed that the iridocorneal angle had been enlarged and macular striae had disappeared.


Influence of Substrates on the Surface Characteristics and Membrane Proteome of Fibrobacter succinogenes S85.
Although Fibrobacter succinogenes S85 is one of the most proficient cellulose degrading bacteria among all mesophilic organisms in the rumen of herbivores, the molecular mechanism behind cellulose degradation by this bacterium is not fully elucidated. Previous studies have indicated that cell surface proteins might play a role in adhesion to and subsequent degradation of cellulose in this bacterium. It has also been suggested that cellulose degradation machinery on the surface may be selectively expressed in response to the presence of cellulose. Based on the genome sequence, several models of cellulose degradation have been suggested. The aim of this study is to evaluate the role of the cell envelope proteins in adhesion to cellulose and to gain a better understanding of the subsequent cellulose degradation mechanism in this bacterium. Comparative analysis of the surface (exposed outer membrane) chemistry of the cells grown in glucose, acid-swollen cellulose and microcrystalline cellulose using physico-chemical characterisation techniques such as electrophoretic mobility analysis, microbial adhesion to hydrocarbons assay and Fourier transform infra-red spectroscopy, suggest that adhesion to cellulose is a consequence of an increase in protein display and a concomitant reduction in the cell surface polysaccharides in the presence of cellulose. In order to gain further understanding of the molecular mechanism of cellulose degradation in this bacterium, the cell envelope-associated proteins were enriched using affinity purification and identified by tandem mass spectrometry. In total, 185 cell envelope-associated proteins were confidently identified. Of these, 25 proteins are predicted to be involved in cellulose adhesion and degradation, and 43 proteins are involved in solute transport and energy generation. Our results supports the model that cellulose degradation in F. succinogenes occurs at the outer membrane with active transport of cellodextrins across for further metabolism of cellodextrins to glucose in the periplasmic space and inner cytoplasmic membrane.


Induction of mucosal and systemic neutralizing antibodies against human immunodeficiency virus type 1 (HIV-1) by oral immunization with bovine Papillomavirus-HIV-1 gp41 chimeric virus-like particles.
Human immunodeficiency virus type 1 (HIV-1) envelope-specific neutralizing antibodies are generated late after initial infection, and the neutralizing antibody response is weak in the infected individuals. Administration of neutralizing antibodies such as 2F5 to HIV-1-infected individuals resulted in reductions in viral loads. Because HIV-1 is transmitted mainly via mucosa and because HIV-specific neutralizing antibodies reduce HIV-1 in infected individuals, a vaccine that can induce both mucosal and systemic HIV-1-specific neutralizing antibodies may be used to prevent and to treat HIV-1 infection. In this study, we made a bovine papillomavirus (BPV) L1-HIV-1 gp41 fusion protein in which ELDKWA of gp41 was inserted into the N terminus of BPV L1 (amino acids 130 to 136). Expression of the fusion protein in insect cells led to the assembly of chimeric virus-like particles (CVLPs). The CVLPs had sizes similar to those of BPV particles and were able to bind to the cell surface and penetrate the cell membrane. Oral immunization of mice with CVLPs induced gp41-specific serum immunoglobulin G (IgG) and intestinal secretory IgA. However, intramuscular immunization with the CVLPs resulted in similar amounts of gp41-specific IgG but low levels of secretory IgA. The antibodies specifically recognized the fixed HIV-1 gp41 on the cell surface. Importantly, the sera and fecal extracts from mice orally immunized with the CVLPs neutralized HIV-1(MN) in vitro. Thus, BPV-HIV-1 gp41 CVLPs may be used to prevent and to treat HIV-1 infection.


Metoclopramide in gastroesophageal reflux disease: rationale for its use and results of a double-blind trial.
We investigated the acute effect of metoclopramide on lower esophageal sphincter pressure, esophageal contraction amplitude, and gastric emptying and compared metoclopramide, 10 mg four times a day, to placebo in improving the symptoms and objective parameters of reflux esophagitis in 19 patients in a randomized, double-blind 4-wk outpatient trial. Orally administered metoclopramide, 10 mg, significantly accelerated gastric emptying of a semisolid meal in patients in whom it was delayed; lower esophageal sphincter pressure was significantly increased for up to 90 min, but there were no changes in esophageal contraction amplitude. During the treatment trial, metoclopramide resulted in an overall improvement in heartburn and regurgitation of 60%, significantly better than 32% improvement after placebo (p less than 0.05). Compared to baseline symptoms scores, metoclopramide significantly improved both daytime and nighttime heartburn and regurgitation. Compared to placebo-treated patients, the metoclopramide group had significantly fewer episodes of daytime heartburn and regurgitation (p less than 0.05), while nighttime symptoms significantly improved with both treatments. Mean antacid consumption was significantly reduced by metoclopramide, 61%, compared to placebo-treated patients, 21% (p less than 0.05), who were ingesting a mean of 1.9 oz of antacid daily. Endoscopic and histological improvement were similar in both groups, although histological healing occurred in three patients after metoclopramide compared with none in the placebo group. Our data suggest that: 1) gastric emptying and lower esophageal sphincter pressure were significantly improved by acute administration of oral metoclopramide; 2) metoclopramide therapy for 4 wk is significantly more effective than placebo (medium dose antacid therapy) in relieving the symptoms of gastroesophageal reflux without significantly altering objective parameters of esophagitis; 3) metoclopramide effectively addresses the diffuse upper gastrointestinal motor disturbances present in reflux esophagitis patients.


[Schisandrin B prevents doxorubicin-induced cardiotoxicity in rabbits].
To study the effect of dexrazoxane in preventing doxorubicin-induced cardiotoxicity in rabbit-models and its mechanism. Thirty one New Zealand white rabbits were randomly divided into two groups, doxorubicin (DOX) group and doxorubicin + dexrazoxane group. The cardiotoxicity was assessed by measuring serum superoxide dismutase (SOD), malondialdehyde (MDA), cardiac troponin I (cTnI), brain natriuretic peptide (BNP), left ventricular ejection function (LVEF), and left ventricular fractional shortening (LVFS), before and 4 and 10 weeks after intervention. Pathological changes in cardiac tissues and the apoptosis of myocardial cells were examined at the end of the experiment. Results Doxorubicin increased serum MDA, cTnI and BNP and decreased SOD, LVEF and LVFS (P < 0.05). Dexrazoxane (DEX) inhibited the increase of MDA, cTnI and BNP, and the decrease of LVEF and LVFS (P < 0.05). The rabbits treated with doxorubicin + dexrazoxane had slighter pathological changes in myocardium and apoptotic myocardial cells than those treated with DOX. Dexrazoxane prevents doxorubicin-induced cardiotoxicity through decreasing oxygen free radical production, cutting down lipid peroxidation, and depressing cardiocyte apoptosis.


Structural basis for Rab11-mediated recruitment of FIP3 to recycling endosomes.
The Rab11 GTPase regulates recycling of internalized plasma membrane receptors and is essential for completion of cytokinesis. A family of Rab11 interacting proteins (FIPs) that conserve a C-terminal Rab-binding domain (RBD) selectively recognize the active form of Rab11. Normal completion of cytokinesis requires a complex between Rab11 and FIP3. Here, we report the crystal structure and mutational analysis of a heterotetrameric complex between constitutively active Rab11 and a FIP3 construct that includes the RBD. Two Rab11 molecules bind to dyad symmetric sites at the C terminus of FIP3, which forms a non-canonical coiled-coiled dimer with a flared C terminus and hook region. The RBD overlaps with the coiled coil and extends through the C-terminal hook. Although FIP3 engages the switch and interswitch regions of Rab11, the mode of interaction differs significantly from that of other Rab-effector complexes. In particular, the switch II region undergoes a large structural rearrangement from an ordered but non-complementary active conformation to a remodeled conformation that facilitates the interaction with FIP3. Finally, we provide evidence that FIP3 can form homo-oligomers in cells, and that a critical determinant of Rab11 binding in vitro is necessary for FIP3 recruitment to recycling endosomes during cytokinesis.


Prediction of one-hour running performance using constant duration tests.
Critical velocity (CV) represents, theoretically, the highest velocity that can be sustained without fatigue. The aim of this study was to compare CV computed from 5 mathematical models in order to determine which CV estimate is better correlated with 1-hour performance and which model provides the most accurate prediction of performance. Twelve trained middle- and long-distance male runners (29 +/- 5 years) performed 3 randomly ordered constant duration tests (6, 9, and 12 minutes), a maximal running velocity test for the estimation of CV, and a 1-hour track test (actual performance). Two linear, 2 nonlinear, and 1 exponential mathematical models were used to estimate CV and to predict the highest velocity that could be sustained during 1 hour (predicted performance). Although all CV estimates were correlated with performance (0.80 < r < 0.93, p < 0.01), it appeared that CV estimated from the exponential model was more closely associated with performance than all other models (r = 0.93; p < 0.01). Analysis of the bias +/- 95% interval of confidence between actual and predicted performance revealed that none of the models provided an accurate prediction of the 1-hour performance velocity. In conclusion, the estimation of CV allows us to rank middle- and long-distance runners with regard to their ability to perform well in long-distance running. However, no models provide an accurate prediction of performance that could be used as a reference for coaches or athletes.


Pars flaccida cholesteatoma as demonstrated by computed tomography.
Prior to the availability of high-resolution computed tomography (CT), the mainstay of roentgenographic diagnosis of cholesteatoma was conventional filming and complex motion tomography. The precise extent of the soft-tissue mass was very difficult to delineate with these methods, and one relied on indirect findings such as bony erosion and displacement. With high-resolution CT, the soft-tissue mass itself can be seen. Typical modes of extention of pars flaccida cholesteatomas of varying size are illustrated in this study.


Applications of Nanosatellites in Constellation: Overview and Feasibility Study for a Space Mission Based on Internet of Space Things Applications Used for AIS and Fire Detection.
In some geographically challenging areas (such as deserts, seas, and forests) where direct connectivity to a terrestrial network is difficult, space communication is the only option. In these remote locations, Internet of Space Things (IoST) applications can also be used successfully. In this paper, the proposed payload for IoST applications demonstrates how an Automatic Identification System (AIS) and a fire detection system can be used effectively. A space mission based on efficient and low-cost communication can use a constellation of nanosatellites to better meet this need. These two applications, which use a constellation of nanosatellites, can provide relevant university-level data in several countries as an effective policy for the transfer of space technology in an educational initiative project. To enhance educational participation and interest in space technology, this paper shares the lessons learned from the project feasibility study based on an in-depth design of a nanosatellite with several analyses (data budget, link budget, power budget, and lifetime estimation). Lastly, this paper highlights by experiments the development and application of a cost-effective sensor node for fire detection and the use of GPS to enable AIS capabilities in the IoST framework.


The Impact of Missing Data and Imputation Methods on the Analysis of 24-Hour Activity Patterns.
The purpose of this study is to characterize the impact of the timing and duration of missing actigraphy data on interdaily stability (IS) and intradaily variability (IV) calculation. The performance of three missing data imputation methods (linear interpolation, mean time of day (ToD), and median ToD imputation) for estimating IV and IS was also tested. Week-long actigraphy records with no non-wear or missing timeseries data were masked with zeros or 'Not a Number' (NaN) across a range of timings and durations for single and multiple missing data bouts. IV and IS were calculated for true, masked, and imputed (i.e., linear interpolation, mean ToD and, median ToD imputation) timeseries data and used to generate Bland-Alman plots for each condition. Heatmaps were used to analyze the impact of timings and durations of and between bouts. Simulated missing data produced deviations in IV and IS for longer durations, midday crossings, and during similar timing on consecutive days. Median ToD imputation produced the least deviation among the imputation methods. Median ToD imputation is recommended to recapitulate IV and IS under missing data conditions for less than 24 h.


Antioxidant effects of a grape seed extract in a rat model of diabetes mellitus.
In the present study we investigated the anti-hyperglycaemic and antioxidant effect of grape seed extract, a polyphenolic flavonoid, in normal and streptozotocin-induced diabetic Wistar rats. Adult male Wistar rats were divided into three groups: Group I: non-diabetic control; Group II: diabetic control; Group III: diabetic rats treated with grape seed extract, administered via an intragastric tube (0.6 ml/rat), at a dose of 100 mg/kg for 20 consecutive days after the induction of diabetes mellitus. Diabetes was induced by an i.p. injection with streptozotocin for groups II and III. TheTBARS, carbonylated proteins, were measured in the plasma and in the supernatant of liver homogenisates, and superoxide dismutase and catalase were measured in the haemolysates of RBCs and supernatant of liver homogenisates. The results showed that oral administration of grape seed extract (100 mg/kg/day) reduced the levels of lipid peroxides and carbonylated proteins and improved the antioxidant activity in plasma and hepatic tissue in rats treated with grape seed natural extract as compared with the diabetic control rats. These results suggested that the grape seed extract enhanced the antioxidant defence against reactive oxygen species produced under hyperglycaemic conditions, hence protecting the liver cells.


Galilean-invariant lattice-Boltzmann models with H theorem.
We demonstrate that the requirement of Galilean invariance determines the choice of H function for a wide class of entropic lattice-Boltzmann models for the incompressible Navier-Stokes equations. The required H function has the form of the Burg entropy for D=2, and of a Tsallis entropy with q=1-(2/D) for D>2, where D is the number of spatial dimensions. We use this observation to construct a fully explicit, unconditionally stable, Galilean-invariant, lattice-Boltzmann model for the incompressible Navier-Stokes equations, for which attainable Reynolds number is limited only by grid resolution.


Imaging After Ureteroscopy: Practice Patterns, Patient Adherence and Impact on Subsequent Management in an Urban Academic Hospital System.
To evaluate practice patterns of post-ureteroscopy (URS) imaging, to assess predictors of imaging order, type and completion, and to analyze impact on patient management. We conducted a retrospective review of patients who underwent URS for nephrolithiasis at a single institution between May, 2020 to May, 2021. Patient demographic, clinical and operative characteristics were reviewed, and surgeons' years in practice. Post-URS imaging studies less than 6 months post-operative were reviewed. Changes in patient management were defined as additional imaging tests ordered or subsequent unplanned surgery. Patient, provider and surgical variables were compared between those who had imaging ordered and those who did not. A total of 289 patients underwent URS. About 234 (81.0%) had post-operative imaging ordered; 147 (62.8%) completed them. Baseline demographics, stone and surgical variables were similar among those who did and did not have imaging ordered and among patients who completed imaging and did not. Pre-operative hydronephrosis was associated with ordering of post-operative imaging (OR = 4.08, P = .01). Urologists in practice less than 5 years were more likely to order post-operative imaging compared to those in practice for more than 5 years (<5: 90.6%, 15+: 53.7%; P <.001). Management changed for 52 of 147 (35.4%) patients who completed imaging; additional imaging was ordered for 38 patients (25.9%) and a second, unplanned surgery was performed for 14 (9.5%). The main predictive factor of ordering post-URS imaging was surgeons' time in practice and pre-operative hydronephrosis. Post-operative imaging changed management in 35.4% of patients. We recommend the development of guidelines encouraging routine imaging for patients following ureteroscopy.


A Wearable Multi-Sensor Array Enables the Recording of Heart Sounds in Homecare.
The home monitoring of patients affected by chronic heart failure (CHF) is of key importance in preventing acute episodes. Nevertheless, no wearable technological solution exists to date. A possibility could be offered by Cardiac Time Intervals extracted from simultaneous recordings of electrocardiographic (ECG) and phonocardiographic (PCG) signals. Nevertheless, the recording of a good-quality PCG signal requires accurate positioning of the stethoscope over the chest, which is unfeasible for a naïve user as the patient. In this work, we propose a solution based on multi-source PCG. We designed a flexible multi-sensor array to enable the recording of heart sounds by inexperienced users. The multi-sensor array is based on a flexible Printed Circuit Board mounting 48 microphones with a high spatial resolution, three electrodes to record an ECG and a Magneto-Inertial Measurement Unit. We validated the usability over a sample population of 42 inexperienced volunteers and found that all subjects could record signals of good to excellent quality. Moreover, we found that the multi-sensor array is suitable for use on a wide population of at-risk patients regardless of their body characteristics. Based on the promising findings of this study, we believe that the described device could enable the home monitoring of CHF patients soon.


Impact of the COVID-19 Pandemic on the Management of Acute Myocardial Infarction.
The coronavirus (COVID-19) outbreak in 2019 has negatively impacted the care of patients with other life-threatening diseases, including acute myocardial infarction (AMI). However, there is little published information concerning the depth of the impact on the clinical management and outcome following AMI. We enrolled patients with AMI who received urgent primary percutaneous coronary intervention at the Beijing Tiantan Hospital (Beijing, China) between December 1, 2019 and April 10, 2020. Patients were divided into 2 cohorts, the pre-COVID-19 group (from December 1, 2019 to January 31, 2020) and during-COVID-19 group (from February 1, 2020 to April 10, 2020) for analysis. The door-to-balloon (D to B) time, total hospitalization stay (days) and coronary care unit (CCU) hospitalization days were calculated. New York Heart Association heart functional class (NYHA class), re-hospitalization and death ratio in patients were assessed between the two cohorts. A total of 148 AMI patients were enrolled in this study comprising 53 patients pre-COVID-19 group and 95 patients during-COVID-19 group. Patients with AMI during-COVID-19 group had longer symptom onset to hospital time (4.5 [2.0-9.3] vs 3.0 [2.0-5.0] hours, p = 0.013) and D to B time (96 [74-119] vs 67 [52-81] minutes, p <0.001); the D to B time shortened during the study period. The two cohorts did not have significantly different number of hospitalization days, re-hospitalization rates, peak cTnI, BNP or death rates. For the one-year follow-up, the patients in the during-COVID-19 group were classified as NYHA class III-IV more frequently (9 [9.7%] vs 0 [0%], p=0.004). The COVID-19 pandemic significantly affected one measure of critical care of patients with AMI, NYHA classification, which may have resulted in increased medical expenses.


Actual proliferating index in oral squamous cell carcinoma and leukoplakia.
To examine the possible association between epithelial proliferation and disease progression in the oral mucosa using the actual proliferation index. The actual proliferation index was measured by the Ki-67 labeling index and argyrophilic nucleolar organizer region count per nucleus. Immunohistochemistry was carried out for Ki-67 by using the molecular immunology borstel-1 clone in 20 leukoplakias, 20 oral squamous cell carcinomas, and 10 normal oral mucosae. The argyrophilic nucleolar organizer region count per nucleus, Ki-67 labeling index, and actual proliferation index were significantly higher in oral squamous cell carcinoma, followed by leukoplakia and normal oral mucosa. Leukoplakia with dysplasia showed a significantly higher Ki-67 labeling index and actual proliferation index, compared to leukoplakia without dysphasia. There was a significant correlation of Bryne's histological malignancy grading with the argyrophilic nucleolar organizer region count and the Ki-67 labeling index. There was a significant positive correlation between the argyrophilic nucleolar organizer region count and the Ki-67 labeling index among all groups. Leukoplakia or suspected epithelial dysplasia should be stained for argyrophilic nucleolar organizer regions and Ki-67. The actual proliferation index is not only useful as a prognostic factor, but could also be a promising treatment determining modality for patients with premalignant and malignant lesions.


Efficacy and safety of carbon-ion radiotherapy for lacrimal gland carcinomas with extraorbital extension: a retrospective cohort study.
To evaluate the efficacy and safety of carbon-ion radiotherapy (CIRT) for patients with lacrimal gland carcinomas (LGCs) with extraorbital extension. The median follow-up period was 53.7 months. The 5-year local control and overall survival rates were 62% and 65%, respectively. Regarding late toxicities, 12 patients (36.4%) developed Grade 4 optic nerve disorders, including visual losses of the diseased side (N = 8; 66.7%), and 1 patient (3.0%) developed a Grade 3 optic nerve disorder. Three patients (9.0%) developed Grade 3 cataracts, 3 (9.0%) developed glaucoma, and 1 (3.0%) developed retinopathy. Two patients (6.1%) had Grade 4 central nervous system necrosis. No Grade 5 late toxicities were observed. The 5-year preservation rate of the ipsilateral eyeball was 86%. Definitive CIRT is effective for LGCs with extraorbital extension with acceptable toxicity. Thirty-three patients treated with CIRT at our institution were analyzed. Sixteen patients (48.5%) had adenoid cystic carcinoma, 8 (24.2%) had adenocarcinoma not otherwise specified, and 9 (27.3%) had other types of the disease. Thirty patients (90.9%) had T4c tumors. The prescribed doses were 57.6 Gy (relative biological effectiveness [RBE]) (N = 18; 54.5%) and 64.0 Gy (RBE) (N = 15; 45.5%) in 16 fractions.


Dynamically tunable narrowband anisotropic total absorption in monolayer black phosphorus based on critical coupling.
A dynamically tunable anisotropic narrowband absorber based on monolayer black phosphorous (BP) is proposed in the terahertz (THz) band. The proposed absorber consists of a monolayer BP and a silicon (Si) grating, which is placed on a silica (SiO2) isolation layer and a gold (Au) substrate. The benefit from the critical coupling mechanism with guided resonance is the efficiency of the absorption can reach 99.9% in the armchair (AC) direction and the natural anisotropy of BP makes it only 87.2% in the zigzag (ZZ) direction. Numerical and theoretical studies show that the absorption efficiency of the structure is operatively controlled by critical coupling conditions, including the geometric parameters of the Si grating, the electron doping of BP and the angle of incident light, etc. More importantly, in the absence of plasmon response, this structure greatly enhances the interaction between light and matter in monolayer BP. In particular, there are several advantages in this structure, such as extremely high-efficiency absorption, excellent tunability, outstanding intrinsic anisotropy and easy manufacturing, which will show unusual and promising potential applications in the design of BP-based tunable high-performance devices.


Effect of controlled local release of sodium fluoride on bone formation: filling a defect in the proximal femoral cortex.
To assess the effect of sodium fluoride (NaF) in the healing of a defect in cortical bone, an experimental model was created by the drilling of 5.0 mm holes in the proximal ends of both femora of 12 adult male New Zealand White rabbits. An interlocking intramedullary implant constructed of poly(d,l-lactic acid) containing NaF was placed in the right femur and an identical implant without NaF (sham), in the left. The implant in the right femur was designed to release NaF in a controlled manner over the duration of the experiment. Ten weeks after implantation, the specimens were removed and were tested in torsion. The mechanical properties were not significantly different between the groups. The femora exposed to NaF had an 18.6% increase in intact cortex near the defect (p = 0.023), however, the deposition of mineralized bone within the defect was not significantly greater. In fact, healing appeared to be impaired by the presence of NaF. There was complete closure of the defect in all but one of the femora with a sham implant, but the tissue had not yet calcified. In contrast, only one femoral defect exposed to NaF had closed. Examination of the material filling the defects of the femora exposed to NaF showed that it was predominantly uncalcified osteogenic mesenchymal tissue.


Different strategies for carboxyl-terminal domain (CTD) recognition by serine 5-specific CTD phosphatases.
The phosphorylated carboxyl-terminal domain (CTD) of RNA polymerase II, consisting of ((1)YSPTSPS(7))(n) heptad repeats, encodes information about the state of the transcriptional apparatus that can be conveyed to factors that regulate mRNA synthesis and processing. Here we describe how the CTD code is read by two classes of protein phosphatases, plant CPLs and yeast Ssu72, that specifically dephosphorylate Ser(5) in vitro. The CPLs and Ssu72 recognize entirely different positional cues in the CTD primary structure. Whereas the CPLs rely on Tyr(1) and Pro(3) located on the upstream side of the Ser(5)-PO(4) target site, Ssu72 recognizes Thr(4) and Pro(6) flanking the target Ser(5)-PO(4) plus the downstream Tyr(1) residue of the adjacent heptad. We surmise that the reading of the CTD code does not obey uniform rules with respect to the location and phasing of specificity determinants. Thus, CTD code, like the CTD structure, is plastic.


Ratio between Epstein-Barr viral load and anti-Epstein-Barr virus specific T-cell response as a predictive marker of posttransplant lymphoproliferative disease.
Posttransplant lymphoproliferative disease (PTLD) is closely linked to primary Epstein-Barr virus (EBV) infection and a defect of EBV specific cellular immunity is supposed to be the basis of PTLD. However, EBV load is so far the only marker proposed to evaluate PTLD risk, and no study has investigated the role of specific anti-EBV T lymphocytes (EBV-TL). We therefore prospectively measured the EBV-TL by enzyme-linked immunospot (elispot) assay, in correlation to EBV load by real-time quantitative PCR and lymphoproliferation occurrence in 45 liver transplanted children. EBV load at the time of primary infection was high in all patients irrespective to subsequent emergence of PTLD. Seven patients developed PTLD, all of them following primary EBV infection. All seven had low EBV-TL (<2/mm3) associated with high viral load (>25,000 copies/microg DNA). Both parameters can be combined in a 100% positive predictive index. Healing from lymphoma was characterized by rapid EBV-TL increase concomitant to decreasing viral load. EBV-TL follow-up helped to adapt immunomodulation. No patient had PTLD whenever EBV-TL were above 2/mm3. We conclude that high viral load is systematic in patients who underwent primary EBV infection and is indicative of the PTLD risk only if there is low concomitant cellular immune response. Healing from PTLD requires modulation of immunosuppression, and appearance of EBV-TL.


[Regenerative medicine in bone tumor surgery].
In bone tumor surgery, target of bone tissue engineering technique will be the reconstruction of large bone void after tumor excision. It may be also useful to accelerate the regeneration of processed autologous bone grafts such as irradiated bone. Interconnected porous ceramics bone substitutes have been considered to be useful as a scaffold for bone engineering. We started a clinical trial on bone regeneration to evaluate its safety and efficiency using mesenchymal stem cells derived from autologous bone marrow aspirates and porous hydroxyapatite ceramics as a scaffold. In this study, after osteoblastic differentiation culture for several weeks, cells integrated to porous ceramics are transplanted into the defect after removal of large benign bone tumor.


Mechanism and consequences of the shift in cardiac arginine metabolism following ischaemia and reperfusion in rats.
Cardiac ischaemia and reperfusion leads to irreversible injury and subsequent tissue remodelling. Initial reperfusion seems to shift arginine metabolism from nitric oxide (NO) to polyamine formation. This may limit functional recovery at reperfusion. The hypothesis was tested whether ischaemia/reperfusion translates such a shift in arginine metabolism in a tumour necrosis factor (TNF)-α-dependent way and renin-angiotensin system (RAS)-dependent way into a sustained effect. Both, the early post-ischaemic recovery and molecular adaptation to ischaemia/reperfusion were analysed in saline perfused rat hearts undergoing global no-flow ischaemia and reperfusion. Local TNF-α activation was blocked by inhibition of TNF-α sheddase ADAM17. To interfere with RAS captopril was administered. Arginase was inhibited by administration of Nor-NOHA. Long-term effects of ischemia/reperfusion on arginine metabolism were analysed in vivo in rats receiving an established ischaemia/reperfusion protocol in the closed chest mode. mRNA expression analysis indicated a shift in the arginine metabolism from NO formation to polyamine metabolism starting within 2 hours (h) of reperfusion and translated into protein expression within 24 h. Inhibition of the TNF-α pathway and captopril attenuated these delayed effects on post-ischaemic recovery. This shift in arginine metabolism was associated with functional impairment of hearts within 24 h. Inhibition of arginase but not that of TNF-α and RAS pathways improved functional recovery immediately. However, no benefit was observed after four months. In conclusion, this study identified TNF-α and RAS to be responsible for depressed cardiac function that occurred a few hours after reperfusion.


Suicidal Ideation among Florida National Guard Members: Combat Deployment and Non-Deployment Risk and Protective Factors.
This study examined relationships among risk/protective factors and suicidal ideation (SI) in deployed and non-deployed National Guard members, particularly examining for possible differential effects of deployment on SI. A total of 3,098 Florida National Guard members completed an anonymous online survey that assessed variables associated with SI including demographics, current psychiatric diagnoses, and pre-, during, and post-deployment experiences. Those deployed had significantly higher rates of SI (5.5%) than those not deployed (3.0%; p < .001). In multivariate analyses, among those not deployed, SI was significantly associated with major depressive disorder (p < .001), posttraumatic stress disorder (PTSD) (p < .001), prior psychological trauma (p < .01), and heavy/hazardous alcohol consumption (p < .05). In contrast, in the deployed, only PTSD (p < .001) and deployment-related mild traumatic brain injury (p < .05) were independently associated with SI. Risk and protective factors differed by deployment status in National Guard members suggesting the possible need for cohort-specific treatment targets to minimize SI.


The Web, communication trends, and children's health. Part 1: Development and technology of the Internet and Web.
The Internet and The World Wide Web are changing the way pediatricians find medical information and provide care. This article, the first in a four-part series on The Web, Communication Trends, and Children's Health, published in consecutive issues of this journal, describes how the Web was created and how its multimedia capabilities are affecting information exchange.


Impacts of incorporating personal genome sequencing into graduate genomics education: a longitudinal study over three course years.
To address the need for more effective genomics training, beginning in 2012 the Icahn School of Medicine at Mount Sinai has offered a unique laboratory-style graduate genomics course, "Practical Analysis of Your Personal Genome" (PAPG), in which students optionally sequence and analyze their own whole genome. We hypothesized that incorporating personal genome sequencing (PGS) into the course pedagogy could improve educational outcomes by increasing student motivation and engagement. Here we extend our initial study of the pilot PAPG cohort with a report on student attitudes towards genome sequencing, decision-making, psychological wellbeing, genomics knowledge and pedagogical engagement across three course years. Students enrolled in the 2013, 2014 and 2015 course years completed questionnaires before (T1) and after (T2) a prerequisite workshop (n = 110) and before (T3) and after (T4) PAPG (n = 66). Students' interest in PGS was high; 56 of 59 eligible students chose to sequence their own genome. Decisional conflict significantly decreased after the prerequisite workshop (T2 vs. T1 p < 0.001). Most, but not all students, reported low levels of decision regret and test-related distress post-course (T4). Each year baseline decisional conflict decreased (p < 0.001) suggesting, that as the course became more established, students increasingly made their decision prior to enrolling in the prerequisite workshop. Students perceived that analyzing their own genome enhanced the genomics pedagogy, with students self-reporting being more persistent and engaged as a result of analyzing their own genome. More than 90% of respondents reported spending additional time outside of course assignments analyzing their genome. Incorporating personal genome sequencing in graduate medical education may improve student motivation and engagement. However, more data will be needed to quantitatively evaluate whether incorporating PGS is more effective than other educational approaches.


Mechanism of biliary secretion of membranous enzymes: bile acids are important factors for biliary occurrence of gamma-glutamyltransferase and other hydrolases.
To elucidate the mechanism of biliary occurrence of gamma-glutamyl transferase [EC 184.108.40.206] and alkaline phosphatase [EC 220.127.116.11], the effect of bile acids on the biliary level of these enzymes was studied in vivo and in vitro. Following intravenous administration of taurocholate, the activities of both enzymes in rat bile increased markedly with a concomitant increase in the excretion of the bile acid. The biliary levels of these enzymes increased to reach a maximum at 10-20 min after administration of the bile acid and decreased thereafter. Right-side-out oriented rat liver canalicular membrane vesicles which localize gamma-glutamyltransferase, aminopeptidase M and alkaline phosphatase on their outer surface (Inoue, M., Kinne, R., Tran, T., Biempica, L., & Arias, I.M. (1983) J. Biol. Chem. 258, 5183-5188) were prepared. Upon incubation of the vesicles with either intact or heat-treated bile samples, the membranous enzymes were released from the vesicles in a time-dependent manner. Incubation of these vesicles with physiological concentrations of taurocholate also solubilized these enzymes from the membranes. Affinity chromatographic analysis on concanavalin A-Sepharose revealed that the transferase thus solubilized retained the hydrophobic domain responsible for anchoring the enzyme to membrane/lipid bilayers. These results indicate that bile acid(s) excreted into the bile canalicular lumen solubilized these enzymes from the apical membrane surface of the biliary tract cells by their detergent action.


[Psychotherapy for depression].
Research on the treatment of depression has shown that both pharmacological and psychosocial interventions are equally efficacious in mild depression as well as moderate and severe depression. Combined psychosocial and pharmacological interventions are also thought to be effective even for severe and chronic depression. Among psychosocial treatments, cognitive behavior therapy, behavior therapy, and interpersonal psychotherapy have been proved to effective. Cognitive behavior therapy is a structured short-term therapy designed to change the patient's negatively distorted cognition. Behavior therapy is a therapy developed based on the understanding that depression is related to a decrease of behaviors that produce positive reinforcement. Interpersonal psychotherapy is a form of short-term therapy to focus on the patient's interpersonal relations with significant others.