Estrogen modulation of G-protein-coupled receptor activation of potassium channels in the central nervous system.
Estrogen rapidly alters the excitability of hypothalamic neurons that are involved in regulating numerous homeostatic functions including reproduction, stress responses, feeding, and motivated behaviors. Neurosecretory neurons, such as gonadotropin-releasing hormone (GnRH) and dopamine neurons, and local circuitry neurons, such as pro-opiomelanocortin (POMC) and gamma-aminobutyric acid (GABA) neurons, are among those involved. We have identified membrane-initiated, rapid-signaling pathways through which 17beta-estradiol (E(2)) alters synaptic responses in these neurons using whole-cell patch recording in hypothalamic slices from ovariectomized female guinea pigs. E(2) rapidly uncouples micro -opioid and GABA(B) receptors from G-protein-gated inwardly rectifying K(+) (GIRK) channels in POMC and dopamine neurons as manifested by a reduction in the potency of micro -opioid and GABA(B) receptor agonists to activate these channels. These effects are mimicked by the selective E(2) receptor modulators raloxifene and 4OH-tamoxifen, the membrane impermeable E(2)-bovine serum albumin (BSA), but not by 17alpha-estradiol. Furthermore, the anti-estrogen ICI 182,780 antagonizes these rapid effects of E(2). Inhibitors of phospholipase C, protein kinase C, and protein kinase A block the actions of E(2), indicating that the E(2) receptor is G-protein-coupled to activation of this cascade. Conversely, estrogen enhances the efficacy of alpha1-adrenergic receptor agonists to inhibit apamin-sensitive small-conductance, Ca(2+)-activated K(+) (SK) currents in preoptic GABAergic neurons; it does so in both a rapid and sustained fashion. Finally, we observed a direct, steroid-induced hyperpolarization of GnRH neurons. These findings indicate that E(2) can modulate K(+) channels in hypothalamic (POMC, dopamine, GABA, GnRH) neurons that are involved in regulating numerous homeostatic functions through multiple intracellular signaling pathways.


Coat color and morphological hair traits influence on the mechanisms related to the heat tolerance in hair sheep.
The principal focus of this study was to describe how the coat characteristics could affect the heat exchanges in animals managed in a hot environment. The Morada Nova ewes were monitored once a month, during 10 consecutive months, in three commercial flocks. Initially, an analysis was performed to measure the differences regarding the coat color in the thermoregulation mechanisms. The animals were grouped into 4 different groups according to coat tonality, as follows: dark red animals (group 1, N = 23), intermediate red color (group 2, N = 27), light red animals (group 3, N = 30), and white-coated animals (group 4, N = 30). The data were collected from 1100 to 1400 h, after the animals were exposed to 30 min of direct sunlight. The cluster analysis was performed considering the hair structural characteristics such as coat thickness (CT, cm), hair length (HL, mm), hair diameter (HD, m), and number of hairs (NH, hairs per unit area), after that these clusters were compared in relation to thermoregulatory mechanisms that include rectal temperature (RT, °C), respiratory rate (RR, breaths min-1), cutaneous evaporation (CE, °C), and respiratory evaporation (RE, W m-2). The groups were characterized and compared using mean and standard deviation, and the differences between the clusters were compared using the Tukey test with a 5% probability of error. In relation to coat color, no differences were found in groups 1, 2, and 3 regarding the activation of the thermoregulation mechanisms. The most different was observed in the totally white coat that presented different thermoregulatory responses as the highest sweating rate. White-coated animals showed a non-pigmented epidermis, and the hair structure is responsible to promote skin protection as necessary, such as a dense coat (1242.7 hair cm-2), long hair (14.2 mm), and thicker coat (7.38 mm). In red-coated animals, the hair structure favored heat loss to the environment, such as short hairs, less thick coat, and less hairs per square centimeter. All evaluated animals showed the rectal temperature within the reference limits for the ovine species, regardless of the coat color. In the analysis of clusters related to the physical structure of hair, it was possible to observe that the animals with thick hair, short hair, and less dense coat tended to have a higher capacity to eliminate heat through their respiratory rate and showed less intense heat loss by cutaneous evaporation. We verified that coat color presents a direct influence on the hair structure and the activation of mechanisms related to thermoregulation.


Variables affecting body burdens of lead, zinc and cadmium in a roadside population of the snailCepaea hortensis Müller.
Application of multiple regression analysis to the lead, zinc and cadmium contents of 985Cepaea hortensis collected from a suburban roadside site over two years showed that body weight, age and daylength were the major factors affecting soft tissue body burdens. Other highly significant influences were physiological changes at the juvenile-adult transition, vapour pressure deficit for the two days before sampling, and mean rainfall for the ten days before. Together these factors accounted for 69%, 79% and 87% of the total variance in lead, zinc and cadmium body burdens respectively. Unexplained variance was provisionally ascribed to individual differences in recent diet and digestive activity. The digestive gland contined the majority of the total body burden of each metal and played an important role in lead and zinc elimination. Whereas levels of lead inC. hortensis fluctuated rapidly, zinc was exchanged more slowly, perhaps by active regulation, and cadmium was effectively immobile, accumulating progressively with age. The implications of these results to the predators of land snails, and to terrestrial monitoring programmes, are discussed.


Steady-state serum concentration of alpha tocopherol not altered by supplementation with oral beta carotene. The Polyp Prevention Study 1 Group.
The antioxidants beta carotene and vitamin E may play a role in cancer prevention. However, some studies have suggested that oral supplements of beta carotene may cause a decrease in serum levels of alpha tocopherol (vitamin E). We conducted this study to determine if beta carotene supplements affect serum levels of vitamin E and vice versa. Five hundred five patients in a clinical trial of antioxidant vitamins, used to prevent recurrences of colonic polyps, received either a placebo, 25 mg of beta carotene per day, 1 g ascorbic acid plus 400 mg alpha tocopherol per day, or all three agents combined. Serum levels of beta carotene and vitamin E were measured before and after 9 months of supplementation, using high-performance liquid chromatography. Vitamin E levels changed very little among the groups receiving placebo or beta carotene and went up substantially and equally in the groups receiving vitamin E plus ascorbic acid or all three agents together. Conversely, beta carotene levels changed very little for the groups receiving placebo or ascorbic acid plus vitamin E but went up substantially and equally for the groups receiving beta carotene alone or all three agents. We conclude that oral supplementation with beta carotene for 9 months does not alter serum concentration of vitamin E and that supplementation with vitamin E plus ascorbic acid does not alter serum beta carotene levels.


Review of the Effects of Offshore Seismic Surveys in Cetaceans: Are Mass Strandings a Possibility?
Displacement of cetaceans is commonly reported during offshore seismic surveys. Speculation concerning possible links between seismic survey noise and cetacean strandings is available for a dozen events but without convincing causal evidence. This lack of evidence should not be considered conclusive but rather as reflecting the absence of a comprehensive analysis of the circumstances. Current mitigation guidelines are inadequate for long-range effects such as displacements and the potential for strandings. This review presents the available information for ten documented strandings that were possibly linked to seismic surveys and recommends initial measures and actions to further evaluate this potential link.


Quality of life and protein-energy wasting in kidney transplant recipients.
Chronic kidney disease has profound effects on the health-related quality of life (HRQoL) of patients, with serious physiological, psychological and socio-economic implications. The co-occurrence of protein-energy wasting and inflammation in end-stage renal disease patients is associated with worse HRQoL and increased mortality. We designed this study to examine the relationship between nutritional and inflammatory status and HRQoL in kidney transplant recipients. Data from 100 randomly selected kidney transplant patients were analyzed in a cross-sectional survey. Socio-demographic parameters, laboratory results, transplantation-related data, comorbidities, medication and malnutrition-inflammation score (MIS) (Kalantar Score) were tabulated at baseline. Patients completed the Kidney Disease Quality of Life-SF (KDQoL-SF™) self-administered questionnaire. Mean age was 51 ± 13 years, median (interquartile range, IQR) time since transplantation 66 (83) months, 57% were men, and 19% had diabetes. The median (IQR) MIS was 3 (3). The MIS significantly and negatively correlated with almost all HRQoL domains analyzed, and this association remained significant in multivariate linear regression analysis for the log-transformed scores on energy/fatigue (β = -0.059 P < 0.001), bodily pain (β = -0.056 P = 0.004), physical functioning (β = -0.029, P = 0.022) and symptoms/problems (β = -0.023 P = 0.005) domains after statistical correction for age, gender, eGFR, dialysis vintage, Charlson Comorbidity Index and occupational status. Additionally, cubic spline analyses revealed linearly increasing, "dose-response" relationship between almost all domains of KDQoL-SF™ and the MIS. Malnutrition-inflammation score is independently associated with different dimensions of HRQoL in kidney transplant recipients.


Barnacle muscle: Ca2+, activation and mechanics.
In this review, aspects of the ways in which Ca2+ is transported and regulated within muscle cells have been considered, with particular reference to crustacean muscle fibres. The large size of these fibres permits easy access to the internal environment of the cell, allowing it to be altered by microinjection or microperfusion. At rest, Ca2+ is not in equilibrium across the cell membrane, it enters the cell down a steep electrochemical gradient. The free [Ca2+] at rest is maintained at a value close to 200 nM by a combination of internal buffering systems, mainly the SR, mitochondria, and the fixed and diffusible Ca(2+)-binding proteins, as well as by an energy-dependent extrusion system operating across the external cell membrane. This system relies upon the inward movement of Na+ down its own electrochemical gradient to provide the energy for the extrusion of Ca2+ ions. As a result of electrical excitation, voltage-sensitive channels for Ca2+ are activated and permit Ca2+ to enter the cell more rapidly than at rest. It has been possible to determine both the amount of Ca2+ entering by this step, and what part this externally derived Ca2+ plays in the development of force as well as in the free Ca2+ change. The latter can be determined directly by Ca(2+)-sensitive indicators introduced into the cell sarcoplasm. A combination of techniques, allowing both the total and free Ca2+ changes to be assessed during electrical excitation, has provided valuable information as to how muscle cells buffer their Ca2+ in order to regulate the extent of the change in the free Ca2+ concentration. The data indicate that the entering Ca2+ can only make a small direct contribution to the force developed by the cell. The implication here is that the major source of Ca2+ for contraction must be derived from the internal Ca2+ storage sites within the SR system, a view reinforced by caged Ca2+ methods. The ability to measure the free Ca2+ concentration changes within a single cell during activation has also provided the opportunity to analyse, in detail, the likely relations between free Ca2+ and the process of force development in muscle. The fact that the free Ca2+ change precedes the development of force implies that there are delays in the mechanism, either at the site of Ca2+ attachment on the myofibril, or at some later stage in the process of force development that were not previously anticipated.(ABSTRACT TRUNCATED AT 400 WORDS)


Simultaneous Targeting of Bladder Tumor Growth, Survival, and Epithelial-to-Mesenchymal Transition with a Novel Therapeutic Combination of Acetazolamide (AZ) and Sulforaphane (SFN).
Current chemotherapies for advanced stage metastatic bladder cancer often result in severe side effects, and most patients become drug resistant over time. Thus, there is a need for more effective therapies with minimal side effects. The acid/base balance in tumor cells is essential for tumor cell functioning. We reasoned that simultaneous targeting of pH homeostasis and survival pathways would improve therapeutic efficacy. We evaluated the effectiveness of targeting pH homeostasis with the carbonic anhydrase inhibitor acetazolamide (AZ) in combination with the survival pathway targeting isothiocyanate sulforaphane (SFN) on the HTB-9 and RT112(H) human bladder tumor cell lines. We assessed viability, proliferation, and survival in vitro and effect on xenografts in vivo. Combination AZ + SFN treatment induced dose-dependent suppression of growth, produced a potent anti-proliferative and anti-clonogenic effect, and induced apoptosis through caspase-3 and PARP activation. The anti-proliferative effect was corroborated by significant reductions in Ki-67, pHH3, cyclin D1, and sustained induction of the cell cycle inhibitors, p21 and p27. Both active p-Akt (Ser473) and p-S6 were significantly downregulated in the AZ + SFN combination treated cells with a concomitant inhibition of Akt kinase activity. The inhibitory effects of the AZ + SFN combination treatment showed similar efficacy as the dual PI3K/mTOR pathway inhibitor NVP-BEZ235, albeit at an expected higher dose. In terms of the effect on the metastatic potential of these bladder cancers, we found downregulated expression of carbonic anhydrase 9 (CA9) concomitant with reductions in both E-cadherin, N-cadherin, and vimentin proteins mitigating the epithelial-to-mesenchymal transition (EMT), suggesting negation of this program. We suggest that reductions in these components could be linked with downregulation of the survival mediated Akt pathway and suggested an active role of the Akt pathway in bladder cancer. Altogether, our in vitro and pre-clinical model data support the potential use of an AZ + SFN combination for the treatment of bladder cancer.


Interaction of rifabutin with model membranes.
Liposomal and free rifabutin were separated by the method of gel filtration. The percents of rifabutin bound to liposomes of different phospholipid composition were measured. The presence of negatively charged phospholipids increased the degree of binding. Binding decreased with increasing the ionic strength. Incubation of rifabutin with liposomes containing anthryl phosphatidylcholine was accompanied by fluorescence quenching. Activity of rifabutin depended on the phospholipid composition of liposomes. Our results indicate that binding of rifabutin is associated with electrostatic and hydrophobic interactions.


Relevance of Zearalenone and its modified forms in bakery products.
Zearalenone is a frequently occurring and well-known mycotoxin developed in cereals before and during the harvest period by Fusarium spp. mainly in maize and wheat. In addition to the main form, various modified forms (phase I and II metabolites) were detected, in some cases in high amounts. These modified forms can be harmful for human health due to their different toxicity, which can be much higher compared to the parent toxin. In addition, the parent toxin can be cleaved from the phase I and II metabolites during digestion. A risk of correlated and additive adverse effects of the metabolites of ZEN phase I and II in humans and animals is evident. ZEN is considered in many studies on its occurrence in grain-based foods and some studies are dedicated to the behavior of ZEN during food processing. This is not the case for the ZEN phase I and II metabolites, which are only included in a few occurrence reports. Their effects during food processing is also only sporadically addressed in studies to date. In addition to the massive lack of data on the occurrence and behavior of ZEN modified forms, there is also a lack of comprehensive clarification of the toxicity of the numerous different ZEN metabolites detected to date. Finally, studies on the fate during digestion of the relevant ZEN metabolites will be important in the future to further clarify their relevance in processed foods such as bakery products.


An overview on the role of bioactive α-glucosidase inhibitors in ameliorating diabetic complications.
Recently the use of bioactive α-glucosidase inhibitors for the treatment of diabetes have been proven to be the most efficient remedy for controlling postprandial hyperglycemia and its detrimental physiological complications, especially in type 2 diabetes. The carbohydrate hydrolysing enzyme, α-glucosidase, is generally competitively inhibited by the α-glucosidase inhibitors and results in the delayed glucose absorption in small intestine, ultimately controlling the postprandial hyperglycemia. Here we have reviewed the most recent updates in the bioactive α-glucosidase inhibitors category. This review provides an overview of the α-glucosidase inhibitory potentials and efficiency of controlling postprandial hyperglycemia of various bioactive compounds such as flavonoids, phenolic compound, polysaccharide, betulinic acid, tannins, anthocyanins, steroids, polyol, polyphenols, galangin, procyanidins, hydroxyl-α-sanshool, hydroxyl-β-sanshool, erythritol, ganomycin, caffeoylquinic acid, resin glycosides, saponins, avicularin, oleanolic acids, urasolic acid, ethanolic extracts etc., from various dietary and non-dietary naturally occurring sources.


A genetic role for macrophage migration inhibitory factor (MIF) in adult-onset Still's disease.
Adult-onset still's disease (AOSD) is a rare systemic inflammatory disorder in which abnormalities in inflammatory cytokines production appear to play a pathophysiological role. Our previous work has reported increased expression of macrophage migration inhibitory factor (MIF) and revealed its correlation with disease severity and activity in AOSD. A -173 G/C single nucleotide polymorphism (SNP) (rs755622) and a -794 CATT₅₋₈ repeat (rs5844572) in the MIF promoter have been reported. In this study, we sought to explore the relationship between functional MIF promoter polymorphisms and MIF expression in AOSD. 100 patients and 200 controls were recruited in the study. A polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay was utilized to analyze the -173 G/C SNP (rs755622) and PCR-based size discrimination assay was applied to detect the -794 CATT₅₋₈ repeat (rs5844572) in the MIF promoter. Plasma MIF levels were measured by ELISA. MIF mRNA levels were quantified by real-time reverse transcription (RT)-PCR. Bisulfate genomic sequencing was employed to evaluate DNA methylation status within the MIF promoter. We identified that the frequencies of MIF -794 CATT₅ (P = 0.001) allele and the expression of MIF (P <0.001) were increased in patients compared to healthy controls. Plasma levels of MIF in patients with CC genotype were higher than those of patients with GC or GG genotypes (P = 0.05). In patients with established AOSD, a higher frequency of -794 CATT₇ containing MIF genotypes was observed in those with liver dysfunction (P = 0.009). Haplotype analysis revealed a higher representation of the MIF haplotype defined by -173*C/-794 CATT₅ (C5) in AOSD patients (P = 0.001). Functional promoter polymorphisms in the MIF gene influence plasma MIF levels in AOSD and may contribute to disease susceptibility or clinical presentation of AOSD.


Calcium signals in neutrophils can be divided into three distinct phases.
Rabbit neutrophils were loaded with the fluorescence probe indo-1 and cytosolic free calcium levels were monitored during chemotactic peptide (fMet-Leu-Phe) stimulation. The fMet-Leu-Phe-induced calcium signal consisted of three consecutive phases: (1) an initial peak that was independent of extracellular calcium, (2) a secondary shoulder that required extracellular calcium but was totally blocked by hyperosmolality and (3) a final plateau of elevated calcium that was dependent on extracellular calcium but insensitive to hyperosmolality.


Mechanisms mediating multiple physiological responses to gonadotropin-releasing hormone.
A central question in endocrinology is how a single ligand interacting with a single receptor can mediate multiple responses. GnRH interaction with receptor offers a prime example, leading to the regulation of synthesis and release of at least three molecules, regulation of target cell responsiveness and receptor number. The present study suggests a molecular model consistent with extant data that provides a mechanism by which this may occur and, further, which allows for coordinate regulation.


[Peripheral nerve block and rebound pain: literature review].
To investigate, describe, and assess the phenomenon of "rebound pain" as a clinically relevant problem in anesthetic practice. The phenomenon of "rebound pain" has been demonstrated and described as a very severe pain, which occurs after a peripheral nerve block resolution with the recovery of sensitivity. The incidence of rebound pain is unknown. Usually, it occurs between 12 to 24hours after surgery and adversely affecting sleep quality. It is not yet possible to establish a mechanism as a definitive cause or trigger factor of rebound pain. Studies suggest that rebound pain is a side effect of peripheral nerve blocks, despite their effectiveness in pain control. Currently, the extent and clinical significance of rebound pain cannot be well determined due to the lack of large prospective studies. Rebound pain assessment should always be considered in clinical practice, as it is not a rare side effect of peripheral nerve blocks. There are still many challenging questions to be answered about rebound pain, so large prospective studies are needed to address the issue. For prevention, the use of peripheral nerve block techniques that avoid nerve damage and adequate perioperative analgesia associated with patient education on the early administration of analgesics, even during the period of analgesia provided by peripheral nerve block, is recommended. A better understanding of the "rebound pain" phenomenon, its pathophysiology, associated risk factors, and long-term consequences may help in developing more effective preventive strategies.


Rotation stability of a toric intraocular lens with a second capsular tension ring.
An Acrysof toric intraocular lens (IOL) and a capsular tension ring (CTR) were implanted in the highly myopic eye of a 74-year-old white man during cataract surgery. On the first postoperative day, the IOL was found 90 degrees from the required position, with a consequent high amount of astigmatism. A second procedure was performed and because it was not possible to secure the toric IOL in the correct position, an additional in-the-bag CTR was inserted, with an immediate optimal outcome. The IOL remained stable up to the final follow-up examination. Co-implantation of a toric IOL and a single CTR has been reported. In our case, 2 CTRs were required to fixate the toric IOL in the correct position. This procedure is simple and safe and should be considered in cases of postoperatively misaligned toric IOLs.


Innervation and serotoninergic receptors of the testis interact with local action of interleukin-1beta on steroidogenesis.
Testosterone secretion by Leydig cells is affected by interleukin-1beta (IL-1beta). The aim of the present study was to investigate whether partial denervation of the testis or local administration of a serotonin (5-HT) receptor antagonist could alter the changes in testicular steoidogenesis induced by IL-1beta. Intratesticular administration of IL-1beta was combined with vasectomy or local injection of ketanserin (5-HT type 2 receptor antagonist) in immature hemicastrated rats and the effect of the interventions on testicular steroidogenesis was studied. One day after treatment with local injection of IL-1beta induced a significant rise in testosterone secretion that could be prevented by vasectomy (that also means transection of the inferior spermatic nerve). In a model in which neither IL-1beta nor ketanserin interfered with steroidogenesis, administration of the receptor antagonist just prior to IL-1beta treatment significantly reduced testosterone secretion. Data indicate interaction between testicular nerves and IL-1beta action and interaction between testicular 5-HT2 receptors and local effect of IL-1beta on testosterone secretion.


Osteochondritis dissecans of the capitellum in fraternal twins: case report.
The etiology of osteochondritis dissecans (OCD) of the capitellum is unknown but has generally been attributed to repetitive microtrauma or ischemia. We present a case report of fraternal twins with OCD lesions in their nondominant arms without a known history of repetitive injury to the elbow. This report provides evidence to support the idea of genetic predisposition for OCD lesions.


MiR-31 targets HSD17B14 and FSHR, and miR-20b targets HSD17B14 to affect apoptosis and steroid hormone metabolism of porcine ovarian granulosa cells.
Porcine 17-hydroxysteroid dehydrogenase type 14 (HSD17B14) and FSH reporter (FSHR) genes play important roles in the metabolism of steroid hormones and the apoptosis of ovarian granulosa cells (GCs). Our bioinformatics analyses and the dual luciferase reporter assays indicated that porcine miR-20b and miR-31 target the 3'-UTR region of HSD17B14 gene, and miR-31 also targets the 3'-UTR region of FSHR gene. Overexpression of porcine HSD17B14 gene promoted the conversion from estradiol (E2) to estrone (E1) and increased the apoptosis of porcine GCs. Overexpression of miR-20b down-regulated the mRNA and protein expression level of HSD17B14 gene, decreased the concentration of progesterone (P4) and E1, increased E2, as well as reduced apoptosis of GCs. Moreover, overexpression of miR-31 also down-regulated the protein expression level of HSD17B14 gene, decreased the concentration of P4 and E1, and increased E2. However, miR-31 promoted apoptosis of GCs by targeting to the 3'-UTR of porcine FSHR gene. Taken together, we found that both porcine miR-20b and miR-31 target HSD17B14 gene, but miR-31 also targets FSHR gene to regulate the metabolism of steroid hormones and the apoptosis of porcine ovarian GCs. These findings expand the epigenetic regulatory mechanism of porcine miR-31 and miR-20b in ovarian GCs.


Rate of oxygen consumption by isolated articular chondrocytes is sensitive to medium glucose concentration.
Cartilage tissue engineering typically involves the culture of isolated chondrocytes within a scaffold material. The oxygen tension within the engineered tissue is known to be an essential parameter for implant success. This will be sensitive to the oxygen consumption behavior of the embedded chondrocytes, which remains to be characterized. We report that the oxygen consumption of bovine articular chondrocytes is sensitive to glucose deprivation below 2.7 mM, increasing from a basal level of 9.6x10(-16) to <18.4x10(-16) mol/cell.h in 1.3 mM glucose. Further studies examined the influence of selecting high (18.4 mM) or low (5.1 mM) glucose medium on the oxygen tension in 2 mm thick cellular agarose constructs. A relative upregulation of oxygen consumption was observed in constructs cultured in low glucose medium. This resulted in the near-anoxic oxygen concentration of 5 microM oxygen in constructs seeded with 40x10(6) cells/ml, compared to 57 microM in the corresponding high glucose culture. The upregulation of oxygen consumption generally corresponded to the inhibition of glycolysis, which is consistent with the Crabtree phenomenon. Medium osmolarity (316-600 mOsm) had minimal effects on chondrocyte oxygen consumption rate. In conclusion, glucose availability is a critical parameter that regulates the oxygen tension within tissue engineered constructs.


Control of shrub establishment by springtime soil water availability in an annual grassland.
Previous studies have shown that the shrub, Baccharis pilularis spp. consanguinea, invades annual grasslands in the San Francisco Bay region in a sporadic manner. Invasion was shown to be positively correlated with the amount of rainfall received in the spring. Here we show that, although Baccharis seeds are dispersed near the beginning of the winter rainy season, seedling root growth is extremely slow until spring. At this time, cessation of the winter rains and transpiration by the grassland annuals results in drying of the upper soil profile. We conclude that establishment of Baccharis seedlings at our study site usually fails because seedling roots cannot reach depths of permanently moist soil, below the depth of the grass roots, before this soil drought occurs. The continuation of rains into the warmer spring months provides a window of time when favorable temperatures and adequate soil moisture allow shrubs to establish.


Medical decision support and medical informatics education: roots, methods and applications in czechoslovakia and the czech republic.
The paper describes the history of medical informatics in Czechoslovakia and the Czech Republic. It focuses on the topics of medical informatics education and decision support methods and systems. Several conferences held in Czechoslovakia and in the Czech Republic organized in cooperation with IMIA or EFMI are described. Support of European Union and Czech agencies in several European and national projects focused on medical informatics topics highly contributed to medical informatics development in Czechoslovakia and the Czech Republic and to the establishment of the European Center for Medical Informatics, Statistics and Epidemiology as the joint workplace of Charles University in Prague and Academy of Sciences of the Czech Republic in 1994.


Convergence between biological, behavioural and genetic determinants of obesity.
Multiple biological, behavioural and genetic determinants or correlates of obesity have been identified to date. Genome-wide association studies (GWAS) have contributed to the identification of more than 100 obesity-associated genetic variants, but their roles in causal processes leading to obesity remain largely unknown. Most variants are likely to have tissue-specific regulatory roles through joint contributions to biological pathways and networks, through changes in gene expression that influence quantitative traits, or through the regulation of the epigenome. The recent availability of large-scale functional genomics resources provides an opportunity to re-examine obesity GWAS data to begin elucidating the function of genetic variants. Interrogation of knockout mouse phenotype resources provides a further avenue to test for evidence of convergence between genetic variation and biological or behavioural determinants of obesity.


Excreted thiocyanate detects live reef fishes illegally collected using cyanide--a non-invasive and non-destructive testing approach.
Cyanide fishing is a method employed to capture marine fish alive on coral reefs. They are shipped to markets for human consumption in Southeast Asia, as well as to supply the marine aquarium trade worldwide. Although several techniques can be used to detect cyanide in reef fish, there is still no testing method that can be used to survey the whole supply chain. Most methods for cyanide detection are time-consuming and require the sacrifice of the sampled fish. Thiocyanate anion (SCN(-)) is a metabolite produced by the main metabolic pathway for cyanide anion (CN(-)) detoxification. Our study employed an optical fiber (OF) methodology (analytical time <6 min) to detect SCN(-) in a non-invasive and non-destructive manner. Our OF methodology is able to detect trace levels (>3.16 µg L(-1)) of SCN(-) in seawater. Given that marine fish exposed to cyanide excrete SCN(-) in the urine, elevated levels of SCN(-) present in the seawater holding live reef fish indicate that the surveyed specimens were likely exposed to cyanide. In our study, captive-bred clownfish (Amphiprion clarkii) pulse exposed for 60 s to either 12.5 or 25 mg L(-1) of CN(-) excreted up to 6.96±0.03 and 9.84±0.03 µg L(-1) of SCN(-), respectively, during the 28 days following exposure. No detectable levels of SCN(-) were recorded in the water holding control organisms not exposed to CN(-), or in synthetic seawater lacking fish. While further research is necessary, our methodology can allow a rapid detection of SCN(-) in the holding water and can be used as a screening tool to indicate if live reef fish were collected with cyanide.


Assessing the mutagenic activities of smoke from different cigarettes in direct exposure experiments using the modified Ames Salmonella assay.
The Ames assay is useful for evaluating the mutagenic potentials of chemicals, and it has been used to evaluate the mutagenic potential of cigarette smoke (CS). In vitro direct exposure systems have been developed to mimic CS exposure in the human respiratory tract, and the Ames assay has been used with such systems. Ames tests were performed using the Vitrocell(®) direct exposure system in this study. The mutagenic potentials of whole mainstream CS and gas/vapor phase fractions produced by conventional combustible cigarettes under two smoking regimens were compared. Salmonella Typhimurium TA98 and TA100 were used with and without metabolic activation, and the number of revertants induced by exposure to each CS was determined. The amount of smoke particles to which cells were exposed were also determined, and dose-response curves describing the relationships between exposure to smoke particles and the number of revertants induced were plotted. The slopes of linear regressions of the dose-response curves were determined, and the slope for each CS was used as a mutagenic activity index for that CS. A new heated cigarette was also tested and smoke from the heated cigarette had a lower mutagenic activity in TA98 and TA100 with metabolic activation than did the conventional CS. The results indicate that the direct exposure system and the Ames test can be used to determine the mutagenic potentials of CS produced by different cigarettes under different conditions (i.e., using different Salmonella Typhimurium strains with and without metabolic activation, and using different smoking conditions).


Mesenchymal stem cells in knee osteoarthritis treatment: A systematic review and meta-analysis.
Stem cells are considered to be one of the greatest potential treatments to cure degenerative diseases. Stem cells injection for knee osteoarthritis (OA) is still a relatively new treatment and has not yet gained popularity. So, the effectiveness, safety and potential of mesenchymal stem cells (MSCs) for knee OA treatment is worthy to be explored. Explore the effectiveness and safety of mesenchymal stem cells (MSCs) in the treatment of knee osteoarthritis. We collected clinical trials using MSCs as treatment for knee OA (before April 2019), including randomized controlled trials (RCTs), retrospective studies and cohort studies. We searched PubMed, EMBASE, Cochrane Library, Web of Science and the ClinicalTrials.gov with keywords (Mesenchymal stem cells [MSCs], Knee osteoarthritis, Effectiveness and Safety), and then performed a systematic review and cumulative metaanalysis of all RCTs and retrospective comparative studies. To evaluate the effectiveness and safety of MSC in knee OA treatment, we applied visual analog scale score, Western Ontario and McMaster Universities Osteo-arthritis Index and adverse events. We included 15 RCTs, two retrospective studies and two cohort studies including a total of 584 knee OA patients in this study. We demonstrated that MSC treatment could significantly decrease visual analog scale in a 12-month follow-up study compared with controls (p < 0.001). MSC therapy also showed significant decreases in Western Ontario and McMaster Universities Osteoarthritis Index scores after the 6-month follow-up (p < 0.001). MSC therapy showed no difference compared with controls (p > 0.05) in adverse events. We suggest that MSC therapy could serve as an effective and safe therapy for clinical application in OA treatment. This study provided the best available evidence and a wider perspective to MSCs application in the management of knee OA. MSCs therapy will have great translational potential in the clinical treatment of various degenerative diseases once optimum formula and explicit target population are identified.


Altered Appetitive Conditioning and Neural Connectivity in Subjects With Compulsive Sexual Behavior.
There has been growing interest in a better understanding of the etiology of compulsive sexual behavior (CSB). It is assumed that facilitated appetitive conditioning might be an important mechanism for the development and maintenance of CSB, but no study thus far has investigated these processes. To explore group differences in neural activity associated with appetitive conditioning and connectivity in subjects with CSB and a healthy control group. Two groups (20 subjects with CSB and 20 controls) were exposed to an appetitive conditioning paradigm during a functional magnetic resonance imaging experiment, in which a neutral stimulus (CS+) predicted visual sexual stimuli and a second stimulus (CS-) did not. Blood oxygen level-dependent responses and psychophysiologic interaction. As a main result, we found increased amygdala activity during appetitive conditioning for the CS+ vs the CS- and decreased coupling between the ventral striatum and prefrontal cortex in the CSB vs control group. The findings show that neural correlates of appetitive conditioning and neural connectivity are altered in patients with CSB. The increased amygdala activation might reflect facilitated conditioning processes in patients with CSB. In addition, the observed decreased coupling could be interpreted as a marker for impaired emotion regulation success in this group.


Does the placement of a FRECA gastrostomy at the time of laparoscopic fundoplication impact on outcome?
To assess the performance of a paediatric cohort having a FRECA PEG (FP) placed at the time of laparoscopic fundoplication. This is retrospective study of a single surgeon's experience of laparoscopic fundoplications over a decade. Patient details were retrieved form a Microsoft Excel database and demographic, operative, and performance measures analysed. Of a series of 67 laparoscopic fundoplications, 20 with neurological compromise underwent FP placement at the time of surgery. Mean age was 3.37 years with a male to female ratio of 1.1:1. A size 9 French FRECA was placed in patients less than 10 kg (12) with larger patients (8) having a size 15 device. A Watson anterior wrap was performed in 16 cases with the rest having a Nissen fundoplication. Seven of these cases had pre-existing FPs which were taken down before replacement post fundoplication. Feeding was resumed the next morning except in three with delayed gastric emptying. Other complications (3) were seen but were not PEG related. The median stay for the series was 4 days (SD 3) and patients were followed up for a mean of 684 days. Over this period four patients relapsed and resumed medical treatment. A single mortality occurred in a syndromic 3-year-old a year later from problems unrelated to surgery. FPs were changed to a button device under general anaesthetic 3-24 months following placement. FP placement at the time of laparoscopic fundoplication does not appear to compromise the outcome of surgery. Neither the size of patient nor the type of wrap is an impediment to its placement and the device can be used shortly after surgery in the majority allowing for an early discharge. Complications are infrequent; however, change to a button device within 2 years of initial placement requires general anaesthetic.


Recent Developments in the Study of Social Relationships, Stress Responses, and Physical Health.
This selective review aims to highlight some of the most recent empirical or theoretical advancements in the study of social relationships as buffers against stress and as protective factors against risk for disease, focusing on articles published between 2013 and 2015. The review summarizes novel findings showing that social relationships can protect individuals against negative health outcomes associated with chronic adversity and can be associated with reduced cumulative physiological damage (allostatic load). There is also evidence that some relationships can be a source of stress. Additionally, recent findings concerning the psychological and neurobiological mechanisms of action for social support, the developmental patterning of social stress-buffering and recent experimental studies attempting to change relationships to affect health are also reviewed.


The excitatory effects of the specific benzodiazepine antagonist Ro14-7437, measured intracellularly in hippocampal CA1 cells.
The specific benzodiazepine antagonist, Ro14-7437, in nanomolar concentrations, caused depolarization, increased spontaneous spiking, and conductance decrease when applied to CA1 cells in vitro. These effects were resistant to intracellularly injected Cl- ions or synaptic blockade by TTX, were prevented in Ca2+-free medium, and occurred with or without prior application of midazolam, an inhibitory benzodiazepine. Ca2+-mediated AHPs and Ca2+ spikes in TTX medium were diminished by the blocker, suggesting that Ro14-7437 acted by inhibiting Ca2+-mediated K+ conductance.


In-vitro susceptibility of Nocardia asteroides to 21 beta-lactam antibiotics, in combination with three beta-lactamase inhibitors, and its relationship to the beta-lactamase content.
Twelve strains of Nocardia asteroides were studied for their susceptibility to 21 beta-lactam antibiotics, alone or in combination with clavulanic acid, sulbactam, or BLP 2013, and for their content of beta-lactamase. Eleven strains were shown to have beta-lactamase activities, which gave three different patterns after analytical isoelectric focusing. The only strain with no detectable beta-lactamase was the most susceptible to the antibiotics tested. The other strains containing beta-lactamases showed almost the same susceptibility profile. Clavulanic acid was the only beta-lactamase inhibitor to show a significant synergistic effect when combined with penicillins.


Production of 230Pa by proton irradiation of 232Th at the LANL isotope production facility: Precursor of 230U for targeted alpha therapy.
Uranium-230 (t1/2 = 20.8 d) is an alpha-emitting radionuclide that has potential application in targeted alpha therapy (TAT) of cancer. Its parent isotope 230Pa (t1/2 = 17.4 d), can be produced by proton irradiation of thorium metal targets. Preliminary 230Pa production runs were performed at the Los Alamos National Laboratory Isotope Production Facility (LANL-IPF) using thin thorium metal targets and a proton beam energy of 15-30 MeV, followed by radiochemical separation of 230Pa from the irradiated target matrix. The measured 230Pa production yields were found to exceed the predicted values in most of the experiments that were performed. This data will inform further production efforts for providing 230Pa/230U for clinical trials.


[Age-related characteristics of the thyroid gland rheogram in healthy persons].
The right and left lobes of the thyroid gland were studied rheographically in 35 healthy persons (70 rheograms) of three age groups for the assessment of the blood flow intensity and functional condition of the thyroid gland vessels. It appeared that the circulation intensity through the thyroid gland decreased with the advance of age; there was also a reduction of elasticity of the vascular wall and an increase in the peripheral resistance of the blood flow in the gland.


Bayesian nonparametric estimation of EQ-5D utilities for United States using the existing United Kingdom data.
Valuations of health state descriptors such as EQ-5D or SF6D have been conducted in different countries. There is a scope to make use of the results in one country as informative priors to help with the analysis of a study in another, for this to enable better estimation to be obtained in the new country than analyzing its data separately. Data from 2 EQ-5D valuation studies were analyzed using the time trade-off technique, where values for 42 health states were devised from representative samples of the UK and US populations. A Bayesian non-parametric approach has been applied to predict the health utilities of the US population, where the UK results were used as informative priors in the model to improve their estimation. The findings showed that employing additional information from the UK data helped in the production of US utility estimates much more precisely than would have been possible using the US study data alone. It is very plausible that this method would serve useful in countries where the conduction of large evaluation studies is not very feasible.


Motor commands of facial expressions: the bereitschaftspotential of posed smiles.
Electroencephalographic (EEG) premotor potentials with negative polarity like the Bereitschaftspotential (BP) are known to precede self-paced voluntary movements of the limbs and other body parts. This is however the first report of such premotor potentials before posed smiles. Scalp EEG was recorded in 16 healthy participants performing self-paced unilateral and bilateral smiles and unilateral finger movements. Amplitudes over six central electrodes and voltage distributions over the entire scalp were compared across conditions at time of EMG-onset, thus focusing on the late BP. Results show the presence of a premotor potential before posed smiles with a later onset, symmetrical bilateral distribution, and smaller amplitude at time of movement-onset, compared to finger movements. Future studies should investigate the BP before various types of emotional and non-emotional facial expressions.


An autosomal recessive syndrome of nasal anomalies associated with renal and anorectal malformations.
We report a consanguineous Arab family with four children affected with bifid nose associated with renal agenesis and variable degree of anorectal malformations. We suggest that the combination of abnormalities in these children represent a previously undescribed autosomal recessive syndrome.


Prediction of highly pathogenic avian influenza vaccine efficacy in chickens by comparison of in vitro and in vivo data: A meta-analysis and systematic review.
Vaccines for avian influenza (AI) can protect poultry against disease, mortality, and virus transmission. Numerous factors, including: vaccine platform, immunogenicity, and relatedness to the field strain, are known to be important to achieving optimal AI vaccine efficacy. To better understand how these factors contribute to vaccine protection, a systematic meta-analysis was conducted to evaluate efficacy data for vaccines in chickens challenged with highly pathogenic (HP) AI. Data from a total of 120 individual trials from 25 publications were selected and evaluated. Two vaccine criteria were evaluated for their effects on two metrics of protection. The vaccine criteria were: 1) the relatedness of the vaccine antigen and challenge strain in the hemagglutinin 1 domain (HA1) protein sequence; 2) vaccine-induced antibody titers to the challenge virus (VIAC). The metrics of protection were: A) survival of vaccinated chickens vs unvaccinated controls; and B) reduction in oral virus-shedding by vaccinated vs unvaccinated controls 2-4 days post challenge. Three vaccine platforms were evaluated: oil-adjuvanted inactivated whole AI virus, recombinant herpes virus of turkeys (rHVT) vectored, and a non-replicating alpha-virus vectored RNA particle (RP) vaccine. Higher VIAC correlated with greater reduction of virus-shed and vaccine efficacy by all vaccine platforms. Both higher HA1 relatedness and higher VIAC using challenge virus as antigen correlated with better survival by inactivated vaccines and rHVT-vectored vaccines. However, rHVT-vectored and RP based vaccines were more tolerant of variation in the HA1; the relatedness of the HA1 of the vaccine and challenge virus did not significantly correlate with survival with rHVT-vectored vaccines. Protection was achieved with the lowest aa similarity for which there was data, 90-93 % for rHVT vaccines and 88 % for the RP vaccine.


Spigelian hernia. A rare case of bilateral hernia and presentation of our experience.
Spigelian hernia is a relatively rare defect of the abdominal wall. Due to its unspecific and variable clinical presentation, presents a difficult diagnostic challenge. We present a rare case of bilateral Spigelian hernia that was diagnosed and successfully treated with open surgery and hernioplasty. Also we present our experience in management of the disease. In our department the last fifteen years, 12 cases of Spigelian hernia have been treated surgically, using direct reconstruction with excellent results.


Skeletal muscle is sensitive to the tension-time integral but not to the rate of change of tension, as assessed by mechanically induced signaling.
Mechanical forces regulate many cellular processes. Mechanotransduction, however, is poorly understood. In skeletal muscle, mechanical forces have a major impact on the regulation of cellular volume, yet the nature of the mechanical stimulation to which muscle is most sensitive is unknown. It was recently demonstrated that activation of the mechanically-sensitive kinase p54 jun-N-terminal-kinase (JNK), is a quantitative marker of mechanical stimulation in skeletal muscle. This marker was shown to be more sensitive to peak tension than to other tension-related parameters such as the tension-time integral (TTI) and the rate of change of tension (dT/dt). The purpose of the present study was to parcel out the contribution of TTI and dT/dt to mechanical stimulation of muscle under conditions of constant peak tension. The rat medial gastrocnemius in situ was subjected to one of four 5-min passive stretch protocols consisting of equal length excursions, but differing in displacement-time integral (4%, 40%, or 100%) and/or rate of stretch (0, 3, or 30 mm/s), and the resulting p54-JNK phosphorylation was assessed. A linear relationship between TTI and p54-JNK signaling was observed. However, no effect of dT/dt was observed. It is concluded that peak tension and TTI are necessary parameters for modeling the mechanical stimulus-response of muscle. Additionally, the mechanism of mechanotransduction is sensitive to peak tension and TTI, but not to dT/dt, and thus exhibits spring-like behavior. These findings may contribute to the refinement of disuse atrophy countermeasures.


Transformed ALDH2-/- hepatocytes by ethanol could serve as a useful tool for studying alcoholic hepatocarcinogenesis.
Alcohol is a well-recognized hepatic carcinogen. Alcohol is metabolized into genotoxic acetaldehyde in hepatocytes, which is catalyzed by aldehyde dehydrogenase 2 (ALDH2). The detailed underlying mechanisms of alcohol-related hepatocellular carcinoma (HCC) remains unclear, at least partially, due to the absence of appropriate experimental models. Current studies suggest that rodents are not good models of the most common liver diseases that trigger HCC including alcoholic liver injury. We hypothesize that ethanol could induce transformation of immortalized normal liver cells, which may serve as a versatile tool for studying alcoholic HCC. Besides, we believe that knockout of ALDH2 will help to shorten the time course of transformation, as ALDH2 deficiency will significantly increase the accumulation of acetaldehyde in hepatocytes. Using this model, the dynamic changes of carcinogenesis-related molecular events could be easily examined. Furthermore, the transformed cells isolated from soft agar could be inoculated to mice for studying invasion, metastasis, and also for screening prophylactics.


Drug analysis by capillary electrophoresis and laser-induced fluorescence.
This review briefly presents the different laser-induced fluorescence detectors, outlines the different dyes used to derivatize molecules which are used with capillary electrophoresis/laser-induced fluorescence (CE-LIF), and provides an overview and current status of CE-LIF in drug analysis.


Chronotherapy improves blood pressure control and reduces vascular risk in CKD.
In patients with chronic kidney disease (CKD), the prevalence of increased blood pressure during sleep and blunted sleep-time-relative blood pressure decline (a nondipper pattern) is very high and increases substantially with disease severity. Elevated blood pressure during sleep is the major criterion for the diagnoses of hypertension and inadequate therapeutic ambulatory blood pressure control in these patients. Substantial, clinically meaningful ingestion-time-dependent differences in the safety, efficacy, duration of action and/or effects on the 24 h blood pressure pattern of six different classes of hypertension medications and their combinations have been substantiated. For example, bedtime ingestion of angiotensin-converting-enzyme inhibitors and angiotensin-receptor blockers is more effective than morning ingestion in reducing blood pressure during sleep and converting the 24 h blood pressure profile into a dipper pattern. We have identified a progressive reduction in blood pressure during sleep--a novel therapeutic target best achieved by ingestion of one or more hypertension medications at bedtime--as the most significant predictor of decreased cardiovascular risk in patients with and without CKD. Recent findings suggest that in patients with CKD, ambulatory blood pressure monitoring should be used for the diagnosis of hypertension and assessment of cardiovascular disease risk, and that therapeutic strategies given at bedtime rather than on awakening are preferable for the management of hypertension.


The PPAR trio: regulators of myocardial energy metabolism in health and disease.
Common causes of heart failure are associated with derangements in myocardial fuel utilization. Evidence is emerging that metabolic abnormalities may contribute to the development and progression of myocardial disease. The peroxisome proliferator-activated receptor (PPAR) family of nuclear receptor transcription factors has been shown to regulate cardiac fuel metabolism at the gene expression level. The three PPAR family members (alpha, beta/delta and gamma) are uniquely suited to serve as transducers of developmental, physiological, and dietary cues that influence cardiac fatty acid and glucose metabolism. This review describes murine PPAR loss- and gain-of-function models that have shed light on the roles of these receptors in regulating myocardial metabolic pathways and have defined key links to disease states including the hypertensive and diabetic heart.


Effects of intracortical infusion of anticholinergic drugs on neuronal plasticity in kitten striate cortex.
During a critical period of postnatal development the mammalian visual cortex is highly susceptible to experience-dependent alterations of neuronal response properties. These modifications are facilitated by the neuromodulators noradrenaline and acetylcholine. To identify the cholinergic mechanisms responsible for this facilitation, muscarinic and nicotinic antagonists were infused into the visual cortex of kittens while the animals were subject to monocular deprivation. Subsequently the ocular dominance of cortical cells was assessed by single-unit recording. Ocular dominance changes were suppressed by scopolamine and pirenzepine but not by gallamine, hexamethonium and mecamylamine. This blocking effect was concentration-dependent, and control experiments revealed that it was not due to suppression of neuronal responses to light. It is concluded from these results that acetylcholine facilitates neuronal plasticity in the visual cortex through mechanisms activated by muscarinic M1 receptors.


NMNAT suppresses tau-induced neurodegeneration by promoting clearance of hyperphosphorylated tau oligomers in a Drosophila model of tauopathy.
Tauopathies, including Alzheimer's disease, are a group of neurodegenerative diseases characterized by abnormal tau hyperphosphorylation that leads to formation of neurofibrillary tangles. Drosophila models of tauopathy display prominent features of the human disease including compromised lifespan, impairments of learning, memory and locomotor functions and age-dependent neurodegeneration visible as vacuolization. Here, we use a Drosophila model of frontotemporal dementia with parkinsonism linked to chromosome 17 (FTDP-17), in order to study the neuroprotective capacity of a recently identified neuronal maintenance factor, nicotinamide mononucleotide (NAD) adenylyl transferase (NMNAT), a protein that has both NAD synthase and chaperone function. NMNAT is essential for maintaining neuronal integrity under normal conditions and has been shown to protect against several neurodegenerative conditions. However, its protective role in tauopathy has not been examined. Here, we show that overexpression of NMNAT significantly suppresses both behavioral and morphological deficits associated with tauopathy by means of reducing the levels of hyperphosphorylated tau oligomers. Importantly, the protective activity of NMNAT protein is independent of its NAD synthesis activity, indicating a role for direct protein-protein interaction. Next, we show that NMNAT interacts with phosphorylated tau in vivo and promotes the ubiquitination and clearance of toxic tau species. Consequently, apoptosis activation was significantly reduced in brains overexpressing NMNAT, and neurodegeneration was suppressed. Our report on the molecular basis of NMNAT-mediated neuroprotection in tauopathies opens future investigation of this factor in other protein foldopathies.


Clinical experience with COP-1 in multiple sclerosis.
COP-1 is one of a series of polypeptide preparations developed to stimulate myelin basic protein (MBP), a natural component of the myelin sheath. MBP in Freund's complete adjuvant induces experimental allergic encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). In saline, MBP suppresses EAE. This is the rationale for the use of COP-1 in MS.


EPH receptor/ephrin system: in the quest for novel anti-angiogenic therapies: Commentary on Hassan-Mohamed et al., Br J Pharmacol 171: 5195-5208.
This article is a Commentary on Hassan-Mohamed I, Giorgio C, Incerti M, Russo S, Pala D, Pasquale EB, Zanotti I, Vicini P, Barocelli E, Rivara S, Mor M, Lodola A and Tognolini M (2014). UniPR129 is a competitive small molecule Eph-ephrin antagonist blocking in vitro angiogenesis at low micromolar concentrations. Br J Pharmacol 171: 5195-5208. doi: 10.1111/bph.12669.


Morpho-regulation of ectodermal organs: integument pathology and phenotypic variations in K14-Noggin engineered mice through modulation of bone morphogenic protein pathway.
Ectodermal organs are composed of keratinocytes organized in different ways during induction, morphogenesis, differentiation, and regenerative stages. We hypothesize that an imbalance of fundamental signaling pathways should affect multiple ectodermal organs in a spatio-temporal-dependent manner. We produced a K14-Noggin transgenic mouse to modulate bone morphogenic protein (BMP) activity and test the extent of this hypothesis. We observed thickened skin epidermis, increased hair density, altered hair types, faster anagen re-entry, and formation of compound vibrissa follicles. The eyelid opening was smaller and ectopic cilia formed at the expense of Meibomian glands. In the distal limb, there were agenesis and hyperpigmentation of claws, interdigital webbing, reduced footpads, and trans-differentiation of sweat glands into hairs. The size of external genitalia increased in both sexes, but they remained fertile. We conclude that modulation of BMP activity can affect the number of ectodermal organs by acting during induction stages, influence the size and shape by acting during morphogenesis stages, change phenotypes by acting during differentiation stages, and facilitate new growth by acting during regeneration stages. Therefore during organogenesis, BMP antagonists can produce a spectrum of phenotypes in a stage-dependent manner by adjusting the level of BMP activity. The distinction between phenotypic variations and pathological changes is discussed.


Unusual cause of hemoptysis: A case report.
Hemoptysis is defined as bleeding from the lower respiratory tract. It can be life-threatening and requires urgent investigation and intervention. Common causes of hemoptysis include bronchiectasis, tuberculosis, aspergilomas and malignancy. We present an unusual case of hemoptysis in a young woman with a history of surgery for scoliosis 18 years earlier. A 30-years-old woman was admitted to our institution for recurring hemoptysis since one year. She had a history of scoliosis and had undergone antero-lateral Th7 through Th12 spinal fusion surgery 18 years earlier. The hemoptysis was slight and resolved spontaneously or after empirical antibiotic therapy, and was attributed to bronchitis. Computed tomography revealed spinal rod penetration into the lung resulting in injury, while the caudal edge of the rod migrated into the liver and the joining screws had entered the mediastinum. Hemoptysis was due to penetration of the rod into the lung. The patient underwent extensive surgery, which was successful. The case highlights the need for thorough evaluation of patients with hemoptysis. Every incident of hemoptysis, even if minor, should be promptly investigated, because it can be life-threatening.


Predisposing factors for neuropathic ulcerations in patients with diabetes.
To assess the risk factors the precipitates ulcerations. In this study a group of 100 patients were selected from November 2007 till October 2008 who were coming for treatment at the Diabetes center of Cluj Napoca, Romania for treatment. 50 cases were patients with ulcerations and 50 without any ulceration. Most important results were: (Age, sex type, biological parameters like HDL, total cholesterol, creatinine ESR, polyneuropathy, arteriopathy, diabetic foot type, diabetes duration). We obtained these results using different statical methods. Ulcerations in diabetic neuropathy are caused by a whole combination of several factors which should be monitored by a meticulous examination and careful follow up of diabetic patients. This in turn will be beneficiary for patients as well as economically interesting reduce the amount of money which is used to treat ulcerations.


Visual search pattern during the line quadrisection task in normal subjects.
A common test for the assessment of hemispatial neglect is the line bisection test. A recent modification of the task has been reported wherein subjects place a mark at either 25% or 75% of line length ('quadrisection'). In a previous study, we suggested that line quadrisection might entail iteration of two steps involved in standard line bisection: (1) finding the midpoint of the entire line, and then (2) bisecting the half-line segment between the adjudged midpoint and the line end ipsilateral to the target quadrisection point. The present study investigated eye movement patterns during line quadrisection in order to test this hypothesis. Twenty normal volunteers determined the left or right quadrisection point of a horizontal line while their eye movements were recorded using two-dimensional video-oculography. Analysis revealed that, like the results of previous studies on bisection, most normal subjects fixate the central portion of the line segment before reaching the target area during the quadrisection task. Normal subjects' eye movements during quadrisection performance share a number of features with line bisection and provide support for the two-step processing hypothesis.


Cognitive and Affective-Motivational Factors as Predictors of Students' Home Learning During the School Lockdown.
During the COVID-19 pandemic, students were facing great challenges. Learning was shifted from the classroom to the home of the students. This implied that students had to complete their tasks in a more autonomous way than during regular lessons. As students' ability to handle such challenges might depend on certain cognitive and motivational prerequisites as well as individual learning conditions, the present study investigates students' cognitive competencies as well as affective-motivational factors as possible predictors of coping with this new learning situation at home. The study uses data of Starting Cohort 2 of the German National Educational Panel Study (NEPS). Data of two measurement points are analyzed: Predictors were assessed at the earlier time point, when students (N=1,452; M age=12years, 8months; 53.4% female) mostly attended seventh grade of a secondary school. They completed competence tests in reading as well as mathematics and rated affective-motivational aspects in terms of willingness to exert effort, learning enjoyment, and intrinsic motivation. One and a half years later - during the COVID-19 pandemic and the first period of school closures - the second measurement point took place. Students' parents rated the situation of learning at home with respect to students' coping with the new situation and parents' difficulties to motivate them. Regression analyses controlling for school track, students' gender, and parents' educational level and parental stress revealed that students' reading competencies and their willingness to exert effort were significant predictors of their coping with the new learning situation at home. Moreover, parents reported that boys were more difficult to motivate to learn during this time as compared to girls. Other predictors (e.g., learning enjoyment) turned out to be non-significant when entered simultaneously in the regression analyses. The results point to the importance of children's prerequisites for autonomous learning situations without structuring elements by teachers within the school context.


Arthroplasty during the COVID-19 Pandemic.
This article aims to give a brief outline of the pathogenesis and transmission of SARS-CoV-2 and how to safely navigate an arthroplasty practice during and after the pandemic.


Clinical manifestations of Borrelia infections of the nervous system.
Clinical, treatment and laboratory parameters were analyzed in 46 consecutive Swedish patients with Borrelia infections of the nervous system. The importance of age in the clinical symptoms, the wide spectrum of disease, and the chronic behaviour of the Borrelia infection of the nervous system was stressed, as well as the benefit of high-dose intravenous antibiotics, especially penicillin G. Borrelia infection of the nervous system can imitate other diseases. When associated with meningitis it can mimic psychosomatic disorders, when associated with radiculoneuritis it may imitate herniated discs and when central nervous involvement of the Borrelia infection occurs, it can mimic a non-infectious, thrombotic or haemorrhagic cerebro-vascular disease.


A comparison of serum bactericidal activity and phenotypic characteristics of bacteremic, pneumonia-causing strains, and colonizing strains of Branhamella catarrhalis.
Four blood isolates, 12 pneumonia isolates, and seven colonizing isolates of Branhamella catarrhalis were compared with respect to their ability to grow in normal human serum and in convalescent serum of a patient with B. catarrhalis bacteremia. Disease-causing isolates showed seven of 16 serum-resistant strains (43 percent) compared with one of seven (13 percent) colonizing strains. Bacteremic strains were not more serum-resistant than pneumonia-causing strains. Trypsin zones of inhibition were higher for disease-causing strains. There was no correlation between source of isolation and colistin sensitivity or ability to hemagglutinate red blood cells.


Epithelium-lined cyst of the pretectal region: case report and electron microscope study.
Ultrastructural findings of an epithelium-lined cyst in the left pretectal region are reported. A 38-year-old woman developed a sensory disturbance on her right side and blurring of vision. Computed tomography scans and magnetic resonance images disclosed a round cystic lesion in the left pretectum. Light microscopically, the cyst was found to be lined by a single layer of cuboidal epithelial cells. Electron microscope examination revealed that the epithelial cells of the cyst possessed clear nuclei, abundant tonofilaments, and glycogen granules, featured as well as the usual organellae. The free surface of the epithelial cells had numerous finger-like microvilli with coating materials. These cells were interconnected by well-developed desmosomes and interdigitations, and also possessed basal lamina materials on the basal surface. These cytological features suggest a heterogenous origin of the cells rather than a neuroepithelial one.


Preoperative Stress Test and Postoperative MI in Patients Requiring Lower Extremity Bypass for Critical Limb Ischemia.
Patients with critical limb ischemia (CLI) often require lower extremity bypass surgery for limb salvage. A myocardial infarction (MI) is a major postoperative risk. Our objective is to assess the utility of preoperative stress test in determining patient outcomes. This is a retrospective study utilizing the national Vascular Quality Initiative database. We collected data from 2013-2018 on all patients undergoing lower extremity bypass for CLI and assessed whether or not they had a preoperative stress test. Rates of an MI were then compared between groups of patients who either did not receive a stress test, had a normal stress test or a positive stress test. An MI was distinguished as troponin only and electrocardiogram (EKG)/clinical. Our secondary end point was in-hospital mortality. Univariate and multivariate analysis with the stress test as a covariate was used to determine significance. During this time period, 29,937 bypasses were performed on 27,219 patients. The average age was 67.5 years (±11.09), 66.3% were men, and 17.3% were African American. Risk factors included hypertension (89.5%), diabetes (55.9%), congestive heart failure (20%), coronary artery disease (32.5%), coronary artery bypass graft (22.2%), and percutaneous coronary intervention (21%). 19,108 patients (64.1%) did not undergo the stress test before bypass, 6,830 (22.9%) had a normal stress test, and 2,898 (9.7%) had a positive stress test. Overall rate of an MI was 4%, with 2% being troponin only and 2% EKG/clinical. The positive stress test had a higher rate of troponin only (2.85%) as well as EKG/clinical (3.37%) MI. For every 10 year increase in age, the odds of having a postoperative MI increased by 27% (P < 0.0001). Overall in-hospital mortality was 1.4%. Patients with positive stress tests had a 2.6% mortality compared with normal/not performed at 1.3%. Of the patients who died, 21.5% had an EKG/clinical MI. Of those patients, 50% did not have a stress test, 12% had normal stress tests, and 23% had positive stress tests. When comparing rates of patients who died or had an MI, there was no difference between patients who had no or a normal stress test (7.29%) versus those who had a positive stress test (7.58%), (P = 0.11). A positive stress test before lower extremity bypass is a significant predictor of a postoperative MI. However, mortality increase was minimal in patients with a positive stress test. Therefore, the stress test result should not delay care for patients needing urgent revascularization.


Inpatient vs outpatient management of pregnancies with vasa previa: A historical cohort study.
Vasa previa, a condition where unprotected fetal blood vessels lie in proximity to the internal cervical opening, is a potentially lethal obstetric complication. The precarious situation of these vessels increases the risk of fetal hemorrhage with spontaneous or artificial rupture of membranes, frequently causing fetal/neonatal demise or severe morbidity. As a result, in many centers, inpatient management forms the mainstay when vasa previa is diagnosed antenatally. This study aimed to determine whether a subpopulation of pregnancies diagnosed antenatally with vasa previa could be safely managed as outpatients. We reviewed all cases of vasa previa in singleton pregnancies, with no fetal anomalies, diagnosed at Mount Sinai Hospital, Toronto, from January 2008 to December 2017. Cases were categorized into three arms for analysis: outpatients (OP), asymptomatic hospitalized (ASH) and symptomatic hospitalized (SH). The SH arm included patients admitted with any antepartum bleeding or suspicious fetal non-stress test. Those that presented with symptomatic uterine activity/threatened preterm labor and delivered within 7 days of diagnosis were excluded from the study. Records were analyzed for details on hospitalization, antenatal corticosteroid administration, cervical length measurements, and fetal/neonatal mortality and morbidity. Of the 84 antenatally-diagnosed cases of vasa previa, 47 fulfilled eligibility criteria. A total of 15 cases were managed as OP, 22 as ASH and 10 as SH. Unplanned cesareans were highest in the SH arm (40% vs. 0% ASH vs. 13.3% OP). Those in the SH arm delivered earliest (median 33.8 weeks, interquartile range (IQR) 33.2-34.3 weeks). Of the asymptomatic patients, those in the ASH arm delivered earlier than those in the OP arm (35.3 [34.6-36.2] weeks vs. 36.7 [35.6-37.2] weeks, p = 0.037). There were no cases of fetal/neonatal death, anemia or severe neonatal morbidity and no significant differences between groups based on cervical length or antenatal corticosteroid administration. Our study suggests that asymptomatic women with an antenatal diagnosis of vasa previa, singleton pregnancies, and at low risk for preterm birth may safely managed as outpatients, as long as they are able to access hospital promptly in the event of antepartum bleeding or early labor.


Pleural liquid pressure in the zone of apposition and in the lung zone.
Pleural liquid pressure in zone of apposition (Pliq,ap) and in lung zone (Pliq,L) was measured simultaneously through liquid filled cannulas in anesthetized dogs in lateral posture. At top Pliq,ap and Pliq,L at iso-height were -9.7 +/- 0.4 and -11.9 +/- 0.3 cm H2O, respectively, at end expiration (eE; P of delta less than 0.01; 11 dogs); -9.6 +/- 0.7 and -19.8 +/- 0.7 cm H2O at end inspiration (eI). At bottom they were -0.6 +/- 0.2 and -2.4 +/- 0.4 cm H2O, respectively, at eE (P of delta less than 0.05; 4 dogs); -1.5 +/- 0.3 and -8.5 +/- 1.0 cm H2O at eI. Vertical gradient of Pliq,ap was -0.97 +/- 0.02 cm H2O/cm. A 132% increase in ventilation (after 10 min dead space breathing) did not change eE and eI Pliq,ap, and eE Pliq,L, but decreased eI Pliq,L. These results imply: (1) no transmission to zone of apposition of tidal changes in Pliq,L; (2) a liquid flow from zone of apposition to lung zone; (3) a net filtration into zone of apposition. They suggest that absorption pressure of capillaries of visceral pleura is greater than that of lymphatics of zone of apposition. Lymphatic role in setting Pliq,L is discussed.


Ranolazine (RS-43285): a preliminary study of a new anti-anginal agent with selective effect on ischaemic myocardium.
Ranolazine (RS-43285) has been shown to possess significant anti-ischaemic properties in a canine model of reversible myocardial ischaemia. The clinical efficacy of this new agent was assessed by a single blind, placebo controlled study of 14 patients with chronic stable angina. A 2 week placebo phase was followed by therapy with 30 mg and 60 mg of ranolazine tid for 2 weeks each. Graded, symptom limited treadmill exercise tests were performed at the end of each phase, 1.5 h (AM) and 7 h (PM) after the morning dose. An additional exercise test 1.5 h after the first dose of 30 mg was included to assess the acute dose response. In the AM study, the mean exercise time increased from 6.9 min (placebo) to 7.8 min after the first dose of 30 mg and to 8.2 min and 8.5 min respectively at the end of 30 mg and 60 mg phases. In the PM study, exercise time improved from 6.5 min (placebo) to 8.2 min and 7.8 min respectively at the end of 30 mg and 60 mg phases. The time to onset of angina showed a similar improvement. No significant changes were observed in the resting and peak exercise heart rates and blood pressure. This preliminary study suggests that ranolazine may significantly prolong exercise time in patients with stable coronary artery disease without altering heart rates and blood pressure.


A completeness indicator of gestational and congenital syphilis information in Brazil.
To evaluate the quality of information on gestational syphilis (GS) and congenital syphilis (CS) on the Sistema de Informação de Agravos de Notificação (SINAN-Syphilis Brazil - Notifiable Diseases Information System) by compiling and validating completeness indicators between 2007 and 2018. Overall, care, and socioeconomic completeness scores were compiled based on selected variables, by using ad hoc weights assigned by experts. The completeness scores were analysed, considering the region and area of residence, the pregnant woman's race/colour, and the year of case notification. Pearson's correlation coefficients were used to validate the scores obtained by the weighted average method, compared with the values obtained by principal component analysis (PCA). Most selected variables presented a good or excellent degree of completeness for GS and CS, except for clinical classification, pregnant woman's level of education, partner's treatment, and child's race/colour, which were classified as poor or very poor. The overall (89.93% versus 89.69%) and socioeconomic (88.71% versus 88.24%) completeness scores for GS and CS, respectively, were classified as regular, whereas the care score (GS-90.88%, and CS-90.72%) was good, despite improvements over time. Differences in the overall, care and socioeconomic completeness scores according to region, area of residence, and ethnic-racial groups were reported for syphilis notifications. The completeness scores estimated by the weighted average method and PCA showed a strong linear correlation (> 0.90). The completeness of GS and CS notifications has been improving in recent years, highlighting the variables that form the care score, compared with the socioeconomic scores, despite differences between regions, area of residence, and ethnic-racial groups. The weighted average was a viable methodological alternative easily operationalised to estimate data completeness scores, allowing routine monitoring of the completeness of gestational and congenital syphilis records.


Non-Gaussian noise-optimized intracellular cytosolic calcium oscillations.
We have numerically studied the effect of a particular kind of non-Gaussian colored noise (NGN), characterized by the deviation q from Gaussian noise (q=1), on intracellular cytosolic calcium (Ca(2+)) oscillations. It is found that, as q is increased, the Ca(2+) oscillation regularity increases and reaches a best performance at an optimal q, and then decreases with further increasing q, which represents the occurrence of coherence resonance, i.e., the most regular Ca(2+) oscillations. Similar phenomena occur for different values of noise intensity and correlation time of the NGN. This phenomenon of deviation-optimized Ca(2+) oscillations show that, external non-Gaussian noises of different types can enhance and even optimize the intracellular Ca(2+) oscillations. This result provides new insights into the constructive roles and potential applications of non-Gaussian noises in intracellular cytosolic Ca(2+) oscillations.


Does the Time of Starting Progesterone Luteal Support Affect Embryo Transfer in Long Agonist Protocol Downregulated ICSI Cycles? A Randomized Controlled Trial.
The aim of this study was to compare the effects of starting progesterone (P4) luteal support (LS) on day of egg retrieval (ER) or day of embryo transfer (ET) on the ratio of difficult ET and cycle outcome. This was a RCT ( ClinicalTrials.gov Identifier: NCT03040830) carried out at Mansoura Integrated Fertility Center (MIFC), Mansoura, Egypt, from November 2015 to January 2017. A total of 171 eligible long agonist ICSI cases were randomly allocated on day of ER into group A (86) starting LS as daily IM 100 mg P4 on day of ER and group B (85) starting P4-LS on day of ET. Difficult ET was defined as blood on ET catheter and/or sounding or dilating the cervix. Primary outcome was the overall ratio of difficult ET and ratios on day 3 and 5 ET. Secondary outcome was the ongoing pregnancy rate (OPR) and implantation rate (IR). The results are presented as % for groups A and B respectively: overall difficult ET (44.1, 24.7) (p = 0.009); day 3 difficult ET (23.2, 24.4) (p = 0.45); day 5 difficult ET (62.7, 25.6) (p = 0.001); overall OPR (38.3, 44.7) (p = 0.43); day 3 ET OPR (41.8, 33.3) (p = 0.51); day 5 ET OPR (34.8, 57.5) (p = 0.048); overall IR (20.0, 22.5) (p = 0.62); day 3 ET IR (17.8, 13.4) (p = 0.44); day 5 ET IR (22, 34.1) (p = 0.09). In conclusion, starting P4 luteal support on egg retrieval day is associated with significantly higher ratio of difficult embryo transfer and lower ongoing pregnancy rate and tendency to lower IR in day 5 ET, so starting P4-LS on day of ET is recommended.


[Incorporation of 15N in VLDL and LDL: in vivo synthesis of apolipoprotein B in the post-absorptive and fasting state].
In vivo synthesis of apolipoprotein B 100 (ApoB) was recently determined in man using stable isotopes. With this procedure we analyzed (1) the effect of fasting on synthesis of ApoB from very low density lipoprotein (VLDL) and (2) tracer enrichment in low density lipoprotein (LDL). After a 36-hour fasting period and in the post-absorptive state 4 healthy subjects were given a priming dose (8.7 mumol/kg) of 15N glycine followed by a constant infusion (10 mumol/kg/h for 8 h) to achieve 5% tracer enrichment in the plasma pool of glycine. The K-values, i.e. fractional synthetic rates/hr of ApoB from VLDL were 0.53 +/- 0.26 vs. 0.43 +/- 0.16 (p greater than 0.05). Tracer enrichment in ApoB from LDL at the end of the infusions was 0.19% vs. 1.46% in ApoB from VLDL. The results indicate that (1) in young healthy postabsorptive individuals about 40% of ApoB from VLDL in plasma is synthesized per hour, (2) fasting does not materially affect fractional ApoB synthesis and (3) at 5% 15N enrichment in plasma glycine, tracer enrichment in ApoB from LDL is at the lower limit of detection for the procedure employed.


Feeding of lovastatin to rats increases the activity of the hepatic mitochondrial outer carnitine palmitoyltransferase.
Administration of lovastatin to male, Sprague-Dawley rats by addition of the drug to the normal chow diet caused a two-fold increase in the activity of the hepatic mitochondrial outer carnitine palmitoyltransferase, but lovastatin apparently did not affect the sensitivity of the outer carnitine palmitoyltransferase to inhibition by malonyl-CoA. There was also no effect of lovastatin on the activity of the hepatic mitochondrial inner carnitine palmitoyltransferase. Feeding of cholestyramine to rats did not affect either the mitochondrial outer carnitine palmitoyltransferase or the mitochondrial inner carnitine palmitoyltransferase.


Mesenchymal stem cell isolation and characterization from human spinal ligaments.
Mesenchymal stem cells (MSCs) have a fibroblast-like morphology, multilineage potential, long-term viability and capacity for self-renewal. While several articles describe isolating MSCs from various human tissues, there are no reports of isolating MSCs from human spinal ligaments, and their localization in situ. If MSCs are found in human spinal ligaments, they could be used to investigate hypertrophy or ossification of spinal ligaments. To isolate and characterize MSCs from human spinal ligaments, spinal ligaments were harvested aseptically from eight patients during surgery for lumbar spinal canal stenosis and ossification of the posterior longitudinal ligament. After collagenase digestion, nucleated cells were seeded at an appropriate density to avoid colony-to-colony contact. Cells were cultured in osteogenic, adipogenic or chondrogenic media to evaluate their multilineage differentiation potential. Immunophenotypic analysis of cell surface markers was performed by flow cytometry. Spinal ligaments were processed for immunostaining using MSC-related antibodies. Cells from human spinal ligaments could be extensively expanded with limited senescence. They were able to differentiate into osteogenic, adipogenic or chondrogenic cells. Flow cytometry revealed that their phenotypic characteristics met the minimum criteria of MSCs. Immunohistochemistry revealed the localization of CD90-positive cells in the collagenous matrix of the ligament, and in adjacent small blood vessels. We isolated and expanded MSCs from human spinal ligaments and demonstrated localization of MSCs in spinal ligaments. These cells may play an indispensable role in elucidating the pathogenesis of numerous spinal diseases.


Identification of novel candidate genes leading to sex differentiation in primordial germ cells of Drosophila.
Germline sex determination and differentiation are pivotal processes in reproduction. In Drosophila, sex determination of the germline occurs in primordial germ cells (PGCs), and the sex differentiation of these cells is initiated during embryogenesis. However, the molecular mechanism initiating sex differentiation remains elusive. To address this issue, we identified sex-biased genes using RNA-sequencing data of male and female PGCs. Our research revealed 497 genes that were differentially expressed more than twofold between sexes and expressed at high or moderate levels in either male or female PGCs. Among these genes, we used microarray data of PGCs and whole embryos to select 33 genes, which are predominantly expressed in PGCs compared to the soma, as candidate genes contributing to sex differentiation. Of 497 genes, 13 genes that were differentially expressed more than fourfold between sexes were also selected as candidates. Among the 46 (33 + 13) candidates, we confirmed the sex-biased expression of 15 genes by in situ hybridization and quantitative reverse transcription-polymerase chain reaction (qPCR) analysis. Six and nine genes were predominantly expressed in male and female PGCs, respectively. These results represent a first step toward elucidating the mechanisms that initiate sex differentiation in the germline.


Local erythropoietin signaling enhances regeneration in peripheral axons.
Erythropoietin (EPO) and its receptor (EPO-R), mediate neuroprotection from axonopathy and apoptosis in the peripheral nervous system (PNS). We examined the impact and potential mechanisms of local EPO signaling on regenerating PNS axons in vivo and in vitro. As a consequence of injury, peripheral nerve axons and DRG neurons have a marked increase in the expression of EPO and EPO-R. Local delivery of EPO via conduit over 2 weeks to rat sciatic nerve following crush injury increased the density and maturity of regenerating myelinated axons growing distally from the crush site. In addition, EPO also rescued retrograde degeneration and atrophy of axons. EPO substantially increased the density and intensity of calcitonin gene-related peptide (CGRP) expression within outgrowing axons. Behavioral improvements in sensorimotor function also occurred in rats exposed to near nerve EPO delivery. EPO delivery led to decreased nuclear factor kappaB (NFkB) activation but increased phosphorylation of Akt and STAT3 within nerve and dorsal root ganglia neurons indicating rescue from an injury phenotype. Spinal cord explant studies also demonstrated a similar dose-dependent effect of EPO upon motor axonal outgrowth. Local EPO signaling enhances regenerating peripheral nervous system axons in addition to its known neuroprotection. Exogenous EPO may have a therapeutic role in a large number of peripheral nerve diseases through its impact on regeneration.


Biosynthesis of Trypanosoma brucei variant surface glycoproteins - analysis of carbohydrate heterogeneity and timing of post-translational modifications.
The variant surface glycoproteins from two cloned populations of Trypanosoma brucei brucei which were known to migrate as multiple bands on SDS gels have been studied. The heterogeneity present was located in those oligosaccharide side chains the addition of which is tunicamycin-sensitive. The time required for the trypanosome to synthesize and express a variant surface glycoprotein molecule in vitro was found, from pulse-chase and limited trypsinisation experiments, to be approximately 40 min. In the light of these data, pulse-chase experiments on the two antigens known to have heterogeneity in their oligosaccharide side chains demonstrated that the heterogeneity probably arose by two different mechanisms. Pulse-chase experiments on three different clones of trypanosomes have also been used to investigate the timing of cleavage of the carboxyl-terminal extension, known to be encoded on variant surface glycoprotein mRNA. Similar pulse-chase experiments followed by immunoprecipitation using affinity purified antiserum have been used to investigate the addition of the cross-reacting determinant. The timing of both these events has been discussed in relation to the time necessary for the synthesis and expression of the variant surface glycoprotein on the surface of the trypanosome.


Molecularly Targeted Agents in the Therapy of Pediatric Brain Tumors.
Pediatric brain tumors are the leading cause of cancer-related death in children. Recent advances in sequencing techniques, and collaborative efforts to encode the mutational landscape of various tumor subtypes, have resulted in the identification of recurrent mutations that may present as actionable targets in these tumors. A number of molecularly targeted agents are approved or in development for the treatment of various tumor types in adult patients. Similarly, these agents are increasingly being incorporated into pediatric clinical trials, allowing for a targeted approach to treatment. However, due to the genetic heterogeneity of these tumors, focused clinical trials in pediatric patients are challenging and regulatory hurdles may delay access to therapeutic compounds that are in regular use in adult patients. The tumor site-agnostic clinical development of TRK inhibitors for pediatric solid tumors is a current example of how the combination of genetic testing and innovative clinical trial design can accelerate the clinical development of targeted agents for pediatric patients.


Polygonal non-wetting droplets on microtextured surfaces.
Understanding the interactions between liquids and solids is important for many areas of science and technology. Microtextured surfaces have been extensively studied in microfluidics, DNA technologies, and micro-manufacturing. For these applications, the ability to precisely control the shape, size and location of the liquid via textured surfaces is of particular importance for the design of fluidic-based systems. However, this has been passively realized in the wetting state thanks to the pinning of the contact line, leaving the non-wetting counterpart challenging due to the low liquid affinity. In this work, confinement is imposed on droplets located on well-designed shapes and arrangements of microtextured surfaces. An active way to shape non-wetting water and liquid metal droplets into various polygons ranging from triangles, squares, rectangles, to hexagons is developed. The results suggest that energy barriers in different directions account for the movement of the contact lines and the formation of polygonal shapes. By characterizing the curvature of the liquid-vapour meniscus, the morphology of the droplet is correlated to its volume, thickness, and contact angle. The developed liquid-based patterning strategy under active regulation with low adhesion looks promising for low-cost micromanufacturing technology, DNA microarrays, and digital lab-on-a-chip.


Deep learning model for breast cancer diagnosis based on bilateral asymmetrical detection (BilAD) in digital breast tomosynthesis images.
The purpose of this study was to develop a deep learning model to diagnose breast cancer by embedding a diagnostic algorithm that examines the asymmetry of bilateral breast tissue. This retrospective study was approved by the institutional review board. A total of 115 patients who underwent breast surgery and had pathologically confirmed breast cancer were enrolled in this study. Two image pairs [230 pairs of bilateral breast digital breast tomosynthesis (DBT) images with 115 malignant tumors and contralateral tissue (M/N), and 115 bilateral normal areas (N/N)] were generated from each patient enrolled in this study. The proposed deep learning model is called bilateral asymmetrical detection (BilAD), which is a modified convolutional neural network (CNN) model of Xception with two-dimensional tensors for bilateral breast images. BilAD was trained to classify the differences between pairs of M/N and N/N datasets. The results of the BilAD model were compared to those of the unilateral control CNN model (uCNN). The results of BilAD and the uCNN were as follows: accuracy, 0.84 and 0.75; sensitivity, 0.73 and 0.58; and specificity, 0.93 and 0.92, respectively. The mean area under the receiver operating characteristic curve of BilAD was significantly higher than that of the uCNN (p = 0.02): 0.90 and 0.84, respectively. The proposed deep learning model trained by embedding a diagnostic algorithm to examine the asymmetry of bilateral breast tissue improves the diagnostic accuracy for breast cancer.


Direct involvement of the tumor suppressor p53 in nucleotide excision repair.
The tumor suppressor p53 enhances repair of UVC-induced DNA damage. The comet-NE assay, a conventional alkaline comet assay which includes a nuclear digestion step, was used to examine the effects of p53 on the excision activity of nuclear extracts (NEs). In contrast with untreated NEs, NEs immunodepleted of p53 or NEs of p53-null cells were unable to excise UVC-induced DNA adducts. Introduction of p53 by transfection restored the excision activity to NEs of p53-null cells. Deletion of the N-terminal 99 amino acids and/or the C-terminal 85 amino acids of p53 barely affected the excision activity, whereas further deletion of the C-terminus of p53 by another 10 amino acids completely abolished the excision activity of NEs. Immunostaining following localized UV irradiation was used to examine the effects of p53 on the recruitment of repair proteins for nucleotide excision repair (NER). Although recruitment of XPC occurred regardless of the presence of p53, the recruitment of XPB was p53-dependent. However, p53 with the 95 amino acid deletion at its C-terminus was unable to support this recruitment of XPB. Consistently, intact p53 (but not the C-terminal 95 residue truncated version) was detected in co-immunoprecipitation assays with an anti-XPB antibody. These results support the hypothesis that p53 facilitates NER through direct involvement by protein-protein interactions.


Ultrastructure and lipid composition of detergent-resistant membranes derived from mammalian sperm and two types of epithelial cells.
Lipid rafts are micro-domains of ordered lipids (Lo phase) in biological membranes. The Lo phase of cellular membranes can be isolated from disordered lipids (Ld phase) after treatment with 1 % Triton X-100 at 4 °C in which the Lo phase forms the detergent-resistant membrane (DRM) fraction. The lipid composition of DRM derived from Madin-Darby canine kidney (MDCK) cells, McArdle cells and porcine sperm is compared with that of the whole cell. Remarkably, the unsaturation and chain length degree of aliphatic chains attached to phospholipids is virtually the same between DRM and whole cells. Cholesterol and sphingomyelin were enriched in DRMs but to a cell-specific molar ratio. Sulfatides (sphingolipids from MDCK cells) were enriched in the DRM while a seminolipid (an alkylacylglycerolipid from sperm) was depleted from the DRM. Treatment with <5 mM methyl-ß-cyclodextrin (MBCD) caused cholesterol removal from the DRM without affecting the composition and amount of the phospholipid while higher levels disrupted the DRM. The substantial amount of (poly)unsaturated phospholipids in DRMs as well as a low stoichiometric amount of cholesterol suggest that lipid rafts in biological membranes are more fluid and dynamic than previously anticipated. Using negative staining, ultrastructural features of DRM were monitored and in all three cell types the DRMs appeared as multi-lamellar vesicular structures with a similar morphology. The detergent resistance is a result of protein-cholesterol and sphingolipid interactions allowing a relatively passive attraction of phospholipids to maintain the Lo phase. For this special issue, the relevance of our findings is discussed in a sperm physiological context.


Levobunolol vs timolol for open-angle glaucoma and ocular hypertension.
A group of 162 patients with chronic open-angle glaucoma or ocular hypertension were treated twice daily for up to 15 months with one of the following topical ophthalmic solutions: 0.5% levobunolol, 1% levobunolol, or 0.5% timolol. Overall mean reductions in intraocular pressure were 8 mm Hg for patients receiving 0.5% levobunolol or timolol and 8.2 mm Hg for patients receiving 1% levobunolol. There were no significant differences between levobunolol and timolol in mean reductions in intraocular pressure, percent of patients with adequately controlled intraocular pressure, or life-table estimates of the probability of successful treatment.