Sleep disturbances after acute exposure to alcohol in mothers' milk.
The results of previous research in our laboratory revealed that breast-fed infants experience significantly less active sleep after exposure to alcohol in their mothers' milk than do breast-fed infants not exposed to alcohol. The present study tested the hypothesis that infants would compensate for such reductions if their mothers then refrained from drinking alcohol. To this end, 23 breast-fed infants from 3 to 5 months of age and their mothers were tested on 2 days separated by 1 week. A small, computerized movement detector, an actigraph, was placed on the infants' left ankles to monitor sleep and activity patterning after which they were bottle fed mother's milk alone (control condition) on 1 test day and mother's milk containing 32 mg of ethanol per 100 ml--the average concentration detected in human milk after lactating women drank an acute dose (0.3 g/kg) of alcohol--on the other. The infants' behaviors were monitored for the next 24 h; the first 3.5 h of monitoring on each test day took place at the Monell Center. Consistent with previous findings, infants exhibited significantly less active sleep during the 3.5 h immediately after exposure to alcohol in mothers' milk compared with the control condition; the decrease in active sleep was observed in all but 4 of the infants tested. Compensatory increases in active sleep were then observed in the next 20.5 h, when mothers refrained from drinking alcohol. Although the mechanisms underlying the reduction in sleep remain to be elucidated, these findings demonstrate that short-term exposure to small amounts of alcohol in mothers' milk produces distinctive changes in the infants' sleep-wake patterning.


Design and synthesis of nonpeptide angiotensin II receptor antagonists featuring acyclic imidazole-mimicking structural units.
Extensive molecular modelling studies, including conformational analysis and the comparison of molecular electrostatic potential distributions, wee used to evaluate structural parameters of new antagonists containing acyclic replacements of the N = C-N imidazole region. The synthesis and the biological screening of a series of acyl biphenyltetrazole derivatives were planned and realized to gain an insight into the structure-activity relationships of this unusual class of Angiotensin II antagonists.


Diversity in the Monogenea and Digenea: does lifestyle matter?
If the cestodes are excluded, then the parasitic platyhelminths of fishes divide neatly into the external and monoxenous Monogenea and the internal and heteroxenous Digenea. Both groups have apparently had long associations of coevolution, host switching and adaptation with fishes and have become highly successful in their respective habitats. Current estimates of species richness for the two groups suggest that they may be remarkably similar. Here we consider the nature of the diversity of the Monogenea and Digenea of fishes in terms of richness of species and higher taxa to determine what processes may be responsible for observed differences. The Monogenea includes at least two super-genera (Dactylogyrus and Gyrodactylus) each of which has hundreds of species; no comparable genera are found in the Digenea. Possible reasons for this difference include the higher host specificity of monogeneans and their shorter generation time. If allowance is made for the vagaries of taxonomic 'lumping' and 'splitting', then there are probably comparable numbers of families of monogeneans and digeneans in fishes. However, the nature of the families differ profoundly. Richness in higher taxa (families) in the Digenea is explicable in terms of processes that appear to have been unimportant in the Monogenea. Readily identifiable sources of diversity in the Digenea are: recolonisation of fishes by taxa that arose in association with tetrapods; adoption of new sites within hosts; adoption of new diets and feeding mechanisms; adaptations relating to the exploitation of ecologically similar groups of fishes and second intermediate hosts; and adaptations relating to the exploitation of phylogenetic lineages of molluscs. In contrast, most higher- level monogenean diversity (other than that associated with the subclasses) relates principally to morphological specialisation for attachment by the haptor.


Freeze-etching of unglycerinated tissue dispersions by application of the oil emulsion technique.
The oil emulsion technique has been applied to rat brain and liver tissue for freeze-etching preparation without antifreeze pretreatment. It has been demonstrated that the dispersed tissue exhibits a high degree of vitrification throughout the mostly globular fragments up to at least 30 micrometer in diameter. Signs of insignificant ice crystal formation of 50 nm and less could only rarely be detected. Although the dispersion process seriously affects the tissue organization as a whole, most cellular organelles and the plasma membrane show satisfactory preservation of structure. In larger tissue drops, a local overall structural preservation of the original cellular organization is recognizable. The results obtained demonstrate the applicability of this method to various biological objects for realizing sufficient cryofixation in the absence of cryoprotectants.


Limbal stem cell disease: Treatment and advances in technology.
Anterior segment stem cell technology, due to its already well-defined corneal limbal stem cells with greater ease of evaluation, has been at the forefront of ophthalmic stem cell treatment and technology since 1997. This paper provides an overview of the current standard of care for treatment of limbal stem-cell deficient conditions and reviews recent treatment technologies using ex vivo expansion of cultivated limbal stem cells of the cornea.


Nonlinear stochastic modeling of aphid population growth.
This paper develops a stochastic population size model for the black-margined pecan aphid. Prajneshu [Prajneshu, A nonlinear statistical model for aphid population growth. J. Indian Soc. Agric. Statist. 51 (1998), p. 73] proposes a novel nonlinear deterministic model for aphid abundance. The per capita death rate in his model is proportional to the cumulative population size, and the solution is a symmetric analytical function. This paper fits Prajneshu's deterministic model to data. An analogous stochastic model, in which both the current and the cumulative aphid counts are state variables, is then proposed. The bivariate solution of the model, with parameter values suggested by the data, is obtained by solving a large system of Kolmogorov equations. Differential equations are derived for the first and second order cumulants, and moment closure approximations are obtained for the means and variances by solving the set of only five equations. These approximations, which are simple for ecologists to calculate, are shown to give accurate predictions of the two endpoints of applied interest, namely (1) the peak aphid count and (2) the final cumulative aphid count.


Evaluation of ameliorative effect of two selected plant drugs on experimentally induced arsenic toxicity in sheep.
Chronic arsenic poisoning is one of the serious health hazards in West Bengal, India, and Bangladesh. It occurs due to contaminated subsoil water. The aim of this study is conducted to find out the ameliorative effect of turmeric and P. foetida powder on experimentally induced arsenic toxicity in sheep. Twelve sheep were divided into four groups; groups I, II and III were orally administered with sodium arsenite at 6.6 mg/kg body weight for 133 days; groups I and II animals were treated by turmeric and P. foetida powders respectively at 500 mg/kg dose for the last 49 days; the fourth group was control. Arsenic content was estimated in faeces, urine and wool in every 15 days. Biochemical, haematological, antioxidant parameters and DNA fragmentation were also assessed. Turmeric and P. foetida powder treatment significantly (P < 0.05) increased arsenic elimination through faeces, urine and wool. Haemoglobin content and TEC were decreased in groups I, II and III; however, these were improved significantly (P < 0.05) by turmeric and P. foetida powder treatment. Increased activity of AST, ALT, blood urea nitrogen and plasma creatinine were significantly (P < 0.05) decreased in groups I and II. The reduced SOD and catalase activity were significantly (P < 0.05) restored at the end of the experiment in turmeric and P. foetida-treated groups. The test drugs are found significantly effective not only to eliminate arsenic from the body but also give protection from possible damage caused by arsenic exposure in sheep.


Temporal characterization of femtosecond laser pulses using tunneling ionization in the UV, visible, and mid-IR ranges.
To generalize the applicability of the temporal characterization technique called "tunneling ionization with a perturbation for the time-domain observation of an electric field" (TIPTOE), the technique is examined in the multicycle regime over a broad wavelength range, from the UV to the IR range. The technique is rigorously analyzed first by solving the time-dependent Schrödinger equation. Then, experimental verification is demonstrated over an almost 5-octave wavelength range at 266, 1800, 4000 and 8000 nm by utilizing the same nonlinear medium - air. The experimentally obtained dispersion values of the materials used for the dispersion control show very good agreement with the ones calculated using the material dispersion data and the pulse duration results obtained for 1800 and 4000 nm agree well with the frequency-resolved optical gating measurements. The universality of TIPTOE arises from its phase-matching-free nature and its unprecedented broadband operation range.


A complication of cricothyroid "minitracheostomy"--oesophageal perforation.
Two patients suffered perforation of the oesophagus during attempted placement of a cricothyroid minitracheostomy. We discuss the probable mechanisms and means of preventing such a recurrence.


Type 2 diabetic patients have increased gluconeogenic efficiency to substrate availability, but intact autoregulation of endogenous glucose production.
An autoregulatory mechanism involving a reciprocal relationship between gluconeogenesis and glycogenolysis regulates endogenous glucose production (EGP) in healthy individuals. In type 2 diabetes, fasting hyperglycemia may be due to increased EGP. To examine gluconeogenesis and autoregulation of EGP in type 2 diabetes, 9 type 2 diabetics and 8 healthy controls were studied during a 3-h infusion of 30 micromol/kg/min Na-lactate. The diabetics were also studied during a control infusion of Na-bicarbonate. To standardize levels of glucoregulatory hormones, plasma insulin, growth hormone, and glucagon were clamped at identical levels during the three experiments. Glucagon levels were elevated from basal levels to approximately 330 ng/l when the lactate or bicarbonate infusions were started, in order to mimic the hyperglucagonemia often seen in diabetes. Lactate gluconeogenesis and total EGP were measured by infusions of [6-(3)H] glucose and [U-14C] lactate. In the bicarbonate experiments, hyperglugagonemia increased lactate gluconeogenesis in the diabetic patients from 4.3+/-1.8 to 6.1+/-2.4 micromol/kg/min (p=0.04). EGP did not change significantly (basal EGP: 15.3+/-3.9, EGP at the end of the study: 14.2+/-3.9 micromol/kg/min, p=0.14). During both lactate experiments, plasma lactate increased more than 4-fold. The increase in lactate gluconeogenesis was significantly higher in diabetics than in controls (values obtained at the end of experiments minus basal values: 10.8+/-3.6 versus 6.4+/-3.6 micromol/kg/min, p=0.03). Compared with normal subjects, the diabetic patients had higher EGP values both at basal conditions (p=0.001) and during lactate infusion (p=0.005). Despite augmented gluconeogenesis, EGP did not change during lactate and glucagon infusion in any of the groups (diabetics, basal EGP: 15.4+/-2.7 versus EGP at the end of experiments: 15.6+/-3.6 micromol/kg/min, p>0.30. Controls, basal EGP: 11.8+/-0.8 versus EGP at the end of experiments: 11.6+/-1.9 micromol/kg/min, p>0.30). Although type 2 diabetics have increased EGP and increased lactate gluconeogenesis, the hepatic autoregulation of EGP during increased substrate-induced gluconeogenesis seems to be intact.


Prognostic value of abdominal sonography in necrotizing enterocolitis of premature infants born before 33 weeks gestational age.
The purpose of this study was to assess the prognostic value of abdominal sonography in necrotizing enterocolitis (NEC) in preterm infants with a gestational age less than 33 weeks of gestation, using surgery and/or death as the primary outcome and stenosis as the secondary outcome. A retrospective study of 95 premature infants (mean gestational age: 28.6 weeks), presenting with NEC between January 2009 and November 2011 and who underwent plain abdominal radiography and sonography, was performed. In uni- and multivariate analyses, radiographic and sonographic findings were correlated with complications ('surgery and/or death' and 'stenosis'). Sonographic findings of free intraperitoneal air (odd ratio [OR]=8.0; IC, 1.4-44.2), free abdominal fluid (OR 3.5; IC 1.3-9.4), portal venous gas (OR 3.9; IC, 1.2-12.9), and bowel wall thickening (OR 2.8; IC,1.1-7.2) were significantly associated with surgery and/or death. Intramural gas was significantly correlated (OR=11.8; IC, 1.5-95.8) with intestinal stenosis following NEC. None of the radiographic findings were associated with complications. Abdominal sonography is a reliable tool for the prognostic assessment of NEC in preterm infants.


Bilaterally absent posterior inferior cerebellar artery: case report.
Posterior inferior cerebellar artery (PICA) is one of the cerebellar arteries which originates from the vertebral artery and has the most complex and variable course. The PICA usually originates from the vertebral artery intracranially as a single trunk, however, absent, double trunk, extracranial, and extradural PICA may also exist. In a collection of 50 cerebellar specimens (100 hemispheres) injected with colored gelatin, one case of bilaterally absent PICA was encountered, male aged 59 (causes of the death was not taken into consideration).


Possible modulation of the medullary respiratory rhythm generator by the noradrenergic A5 area: an in vitro study in the newborn rat.
Respiratory activity was recorded on hypoglossal nerve or ventral cervical roots during in vitro experiments performed in the superfused brainstem-cervical cord preparation of newborn rats. Section and coagulation experiments revealed that the medullary respiratory generator was tonically inhibited by a structure located in the caudal ventrolateral pons. Electrical and pharmacological stimulations located this structure more precisely between the superior olivary nuclei and the sensory nucleus of the Vth nerve, i.e. in an area containing the A5 noradrenergic nucleus. Norepinephrine and alpha 2-antagonists (yohimbine, idazoxan) added to the bathing medium modified the respiratory frequency. Norepinephrine decreased respiratory frequency whereas norepinephrine antagonists increased respiratory rate. The electrical stimulation of the caudal ventrolateral pons which inhibited the respiratory rhythm under normal bathing medium became ineffective after alpha 2-antagonist. The results herein suggest that a noradrenergic inhibitory drive, originating from the A5 area or surrounding structures modulates the activity of the medullary respiratory generator. This hypothesis is discussed in relation to A5 involvement in cardiovascular regulation.


Stem cell gene expression in MRPS18-2-immortalized rat embryonic fibroblasts.
We have recently found that primary rat embryonic fibroblasts (REFs) could be immortalized by overexpression of the human mitochondrial ribosomal protein MRPS18-2 (S18-2). A derived cell line, designated 18IM, expressed the embryonic stem cell markers SSEA-1 and Sox2. Upon inoculation into severe combined immunodeficiency mice, 18IM cells differentiated to express pan-keratin. They were not tumorigenic. Here we report the gene profiling of 18IM, compared with REF cells. Pathways involved in oxidative phosphorylation, ubiquinone (Coenzyme Q 10) biosynthesis, fatty acid elongation in mitochondria, PI3K/AKT signaling, a characteristic of rapidly proliferating cells, were upregulated in 18IM. Genes involved in the transcription/translation machinery and redox reactions, like elongation factors, ATP synthases, NADH dehydrogenases, mitogen activated kinases were upregulated as well. 18IM cells produced more pyruvate, indicating enhanced ATP synthesis. The expression of Oct4, Sox2, and Nanog that can contribute to the experimental induction of pluripotency in primary fibroblasts was also elevated, in contrast to Klf4 and C-myc that were downregulated. Subsequently, three new immortalized cell lines were produced by S18-2 overexpression in order to check the representativeness of 18IM. All of them showed anchorage-independent growth pattern. Two of three clones lost vimentin and smooth muscle actin, and expressed Sox2 and Oct4. We suggest that S18-2 is involved in the developmental regulation.


Hydroxy levamisole and its phase II conjugates as potential indicators of levamisole doping in thoroughbred horses.
According to previous research, aminorex is the major metabolite of levamisole; however, in the screening of levamisole-positive racehorse urine and plasma samples, aminorex could only be detected in trace amounts or not at all. In forensic laboratories, hydroxy levamisole and its phase II conjugates make it easier to confirm levamisole misuse and to differentiate between the abuse of levamisole and aminorex. This study aimed to identify the major levamisole metabolites that can be detected along with the parent drug. The study describes levamisole and its metabolites in thoroughbred horses following oral administration and in vitro with equine liver microsomes. The plausible structures of the detected metabolites were postulated using liquid chromatography combined with high-resolution mass spectrometry. Under the experimental conditions 26 metabolites (17 phase I, 2 phase II, and 7 conjugates of phase I metabolites) were detected (M1-M26). The major phase I metabolites identified were formed by hydroxylation. In phase II, the glucuronic acid conjugates of levamisole and hydroxy levamisole were detected as the major metabolites. In plasma, the parent drug and major metabolites are detectable for up to eight days, while in urine, they are detectable for up to twenty days. Levamisole levels rapidly increased at 45 min following administration, then declined gradually until detectable levels were reached approximately 8 days after administration, according to a pharmacokinetics study. A prolonged elimination profile and relatively high concentration of hydroxy metabolites suggest that the detection of hydroxy metabolites is imperative for investigating levamisole doping in horses.


Intravenous pyogenic granuloma of the hand.
Intravenous pyogenic granuloma is a recently described counterpart of cutaneous pyogenic granuloma. We report the second known case of this entity in the orthopedic and hand surgery literature and describe the characteristic clinical and histologic findings associated with this rare benign tumor.


Action of glutamic acid and of some glutamate analogues on the molluscan central neurones.
The effects of L-glutamic acid and of some glutamate analogues have been studied on the central nervous system of the snail Heobania vermiculata, using conventional electrophysiological techniques. The glutamate H-response had the mean equilibrium value of -(57 +/- 4) mV and was associated with a Cl- conductance change. The D-response to glutamate application showed an involvement of sodium ions. Aspartate was agonist of glutamate action and displayed similar equilibrium value of the H-response, whereas quisqualate H-response was 'non-invertible'.


Neural inputs from the uterus to the paraventricular magnocellular neurons in the rat.
Extracellular action potentials were recorded from antidromically identified, tonically firing cells in the hypothalamic paraventricular nucleus (PVN) of ovariectomized, estrogen-treated female rats under urethane anesthesia. Genital or somatic sensory stimuli, or electrical stimulation of the nerves innervating the pelvis were applied. Uterine horn or vaginal distension each excited 33% of the neurons tested. Probing of the cervix had no effect. Hindpaw pinch produced excitation in 39% and inhibition in 11% of the neurons tested. Non-noxious somatic stimuli had no effect. Stimulation of the uterine afferent nerves, the hypogastric and pelvic nerves, excited 55% and 30% of the neurons tested, respectively. Stimulation of a somatic nerve of the hindleg, the sciatic nerve, activated 80% of the neurons tested. These results indicate that specific sensory afferents arrive at the PVN from the uterus; in addition, somatic afferents converge in this hypothalamic nucleus.


Chemical shift and magnetic susceptibility contributions to the separation of intracellular and supernatant resonances in variable angle spinning NMR spectra of erythrocyte suspensions.
Factors contributing to the observation of two separate water resonance arising from erythrocyte suspensions under magic- and variable-angle spinning conditions were examined. By observing the 1H NMR spectra of different chemical species in erythrocytes at different spinning angles, two major effects of comparable magnitude were shown to contribute to the separation: 1) an isotropic chemical shift difference, and 2) a susceptibility difference between the intracellular and supernatant compartments. When the sample was spun at the magic angle, the susceptibility difference did not contribute to the separation. Use of different angles between the spinning axis and the main magnetic field provided a method for quantifiying the isotropic chemical shift and susceptibility differences between the compartments.


Nutrient strengthening of winter wheat by foliar ZnO and Fe3O4 NPs: Food safety, quality, elemental distribution and effects on soil bacteria.
With the anticipated application of engineered nanomaterials (ENMs) as foliar fertilizers in agriculture, there is a particular need to accurately assess crop intensification capacity, potential hazards, and effects on the soil environment when ENMs are applied alone or in combination. In this study, the joint analysis of scanning electron microscopy (SEM), X-ray diffraction (XRD) and vibrating sample magnetometer (VSM) showed that ZnO NPs transformed on the leaf surface or within the leaf, and Fe3O4 NPs were able to translocate from the leaf (~ 25 memu/g) into the stem (~ 4 memu/g), but were unable to enter the grain (below 1 memu/g), guaranteeing food safety. Spray application of ZnO NPs significantly improved grain Zn content of wheat (40.34 mg/kg), whereas Fe3O4 NPs treatment and Zn + Fe NPs treatment did not significantly improve grain Fe content. According to the micro X-ray fluorescence of wheat grains(μ- XRF) and physiological structure in situ analysis showed that ZnO NPs treatment and Fe3O4 NPs treatment could increase the elemental contents of Zn and Fe in the crease tissue and endosperm components, respectively, while antagonism was observed in the grain treated with Zn + Fe NPs. The 16S rRNA gene sequencing results showed that the Fe3O4 NPs treatment had the greatest negative effect on soil bacterial community, followed by Zn + Fe NPs, and ZnO NPs showed some promotion effect. This may be caused by the significantly higher elemental contents of Zn/Fe in the treated roots and soils. This study critically evaluates the application potential and environmental risks of nanomaterials as foliar fertilizers and is instructive for agricultural applications of nanomaterials alone and in combination.


Brain tumor location influences the onset of acute psychiatric adverse events of levetiracetam therapy: an observational study.
To explore possible correlations among brain lesion location, development of psychiatric symptoms and the use of antiepileptic drugs (AEDs) in a population of patients with brain tumor and epilepsy. The medical records of 283 patients with various types of brain tumor (161 M/122 F, mean age 64.9 years) were analysed retrospectively. Patients with grade III and IV glioma, previous history of epileptic seizures and/or psychiatric disorders were excluded. Psychiatric symptoms occurring after initiation of AED therapy were considered as treatment emergent psychiatric adverse events (TE-PAEs) if they fulfilled the following conditions: (1) onset within 4 weeks after the beginning of AED therapy; (2) disappearance on drug discontinuation; (3) absence of any other identified possible concurrent cause. The possible influence of the following variables were analysed: (a) AED drug and dose; (b) location and neuroradiologic features of the tumor, (c) location and type of EEG epileptic abnormalities, (d) tumor excision already or not yet performed; (e) initiation or not of radiotherapy. TE-PAEs occurred in 27 of the 175 AED-treated patients (15.4%). Multivariate analysis showed a significant association of TE-PAEs occurrence with location of the tumor in the frontal lobe (Odds ratio: 5.56; 95% confidence interval 1.95-15.82; p value: 0.005) and treatment with levetiracetam (Odds ratio: 3.61; 95% confidence interval 1.48-8.2; p value: 0.001). Drug-unrelated acute psychiatric symptoms were observed in 4 of the 108 AED-untreated patients (3.7%) and in 7 of the 175 AED-treated patients (4%). The results of the present study suggest that an AED alternative to levetiracetam should be chosen to treat epileptic seizures in patients with a brain tumor located in the frontal lobe to minimize the possible onset of TE-PAEs.


Rh-Catalyzed Coupling of Aldehydes with Allylboronates Enables Facile Access to Ketones.
We present herein a novel strategy for the preparation of ketones from aldehydes and allylic boronic esters. This reaction involves the allylation of aldehydes with allylic boronic esters and the Rh-catalyzed chain-walking of homoallylic alcohols. The key to this successful development is the protodeboronation of alkenyl borylether intermediate via a tetravalent borate anion species in the presence of KHF2 and MeOH. This approach features mild reaction conditions, broad substrate scope, and excellent functional group tolerance. Mechanistic studies also supported that the tandem allylation and chain-walking process were involved.


Perspectives on the development of acyclic nucleotide analogs as antiviral drugs.
The development of Viread (tenofovir disoproxil) for HIV and Hepsera (adefovir dipivoxil) for HBV presented many unique challenges. Unlike nucleosides and most conventional drugs, the parent acyclic nucleotide analogs are charged at physiologic pH and not suitable for oral administration which is highly desired in chronic therapies. Physicochemical properties, cellular permeation, renal toxicity, and bioavailability all had to be addressed during the development of these compounds. As a class, the acyclic nucleotides have long intracellular half-lives, allowing once-daily dosing, which provided the initial rationale for treatment of chronic viral diseases such as HIV and HBV. Prodrugs originally designed to deliver the parent acyclic nucleotide analog to the systemic circulation, also function to increase the tissue distribution and intracellular concentrations of the acyclic nucleotide diphosphate inside cells.


[Particulars on the regulation of Ca(2+) in the cell nucleus].
The data of literature on the structural and functional features of nuclear pore complexes, the role of receptor-activated Ca(2+)-channels and Ca(2+)-ATPase of nuclear membranes are presented. The possible ways of Ca(2+)-signal transmission into the nucleus and the role of nuclear Ca2+ in apoptosis are discussed.


Organization and sequence of five tRNA genes and of an unidentified reading frame in the wheat chloroplast genome: evidence for gene rearrangements during the evolution of chloroplast genomes.
The genes for the initiator tRNA(Met)CAU, tRNA(Gly)UCC, tRNA(Thr)GGU, tRNA(Glu)UUC and tRNA(Tyr)GUA and an open reading frame of 62 codons have been identified by sequencing a 2,358 bp BamHI and a 1,378 bp BamHI-Sst2 DNA fragments from wheat chloroplasts. A comparison of the organization of these five tRNA genes and of the open reading frame on the wheat, tobacco and spinach chloroplast genomes suggests that at least three genomic inversions must have occurred during the evolution of the wheat chloroplast genome from a spinach-like ancestor genome. Furthermore, it seems that in wheat the 91 bp intergenic region between the genes for the initiator tRNA(Met) and the gene for tRNA(Gly)UCC is one end-point of the 20 kbp genomic inversion proposed by Palmer and Thompson in the case of maize (Palmer and Thompson 1982). A 119 bp duplication is located at this junction: the first copy comprises the 91 bp of the intergenic region and the first 28 bp of the tRNA(Met) gene, the second copy is found downstream of the tRNA(Met) gene.


[The conditions of arrangement of the thrombin generation test to detect the hypercoagulation status].
The study covered the impact of modes of preparation of plasma samples to arrange the thrombin generation test for the purpose to establish adequately the hypercoagulation status. The optimal regimen is determined to prepare the samples to be used in the study. The group of females was involved into the study to take the composite oral contraceptives to demonstrate the possibility to apply the thrombin generation test to reveal the hypercoagulation.


Biochemical analysis of meiosis in the male mouse. II. DNA metabolism at pachytene.
The DNA metabolism of mouse spermatogenic cells was investigated by intravenous administration of isotope and Staput fractionation of the cells. The pattern of metabolism is virtually identical with that observed in Lilium microsporocytes. A programmed single strand nicking of DNA occure at pachytene such that at least 50% of the DNA is in the form of 62S fragments. Repair replication of endogenously nicked sites is fully achieved during pachytene. The sites of nicking and repair are preferentially located in sequences that are repeated about 400 times. These results are considered as strong evidence for a universal pattern of meiotic prophase DNA metabolism which is associated with crossing-over.


Does image-guided biopsy of discitis-osteomyelitis provide meaningful information to impact clinical management?
The aims of this study are to assess the diagnostic yield of image-guided biopsy for discitis-osteomyelitis (DO), identify factors associated with biopsy yield (laboratory, pre-defined MRI findings, and biopsy technique), and impact of biopsy on management of patients appropriately selected according to the Infectious Disease Society of America guidelines (IDSA). This is a retrospective review of patients who underwent biopsy for suspected DO from 2011 to 2019. Reference standards to establish diagnosis of DO in order were histopathology/microbiology from biopsy or subsequent surgical sampling, positive blood culture or serology, and imaging/clinical follow-up. Laboratory markers, pre-biopsy antibiotics and MRI features, procedural-related variables, and impact of biopsy on management were assessed. Multivariable logistic regression was also performed. Out of 97 included patients, 78 were diagnosed with DO. Overall sensitivity of biopsy for detecting DO was 41.0% (32/78), including 10 patients with positive histopathology only, 14 with positive biopsy culture only, and 8 with both. Elevated ESR (p < 0.001) and epidural collection on MRI (p = 0.008) were associated with higher biopsy yield (63.6% and 68.6%, respectively) in a multivariable model. Procedural variables were not associated with yield. Biopsy results impacted the management in 19/77 (24.7%) patients, of whom 15/19 (78.9%) had treatment de-escalation and 4/19 (21.0%) had treatment escalation including starting new anti-tuberculous and anti-fungal regimens. Sensitivity of biopsy for detecting DO was 41.0%. When IDSA guidelines are followed, biopsy provided impactful information that changed the management in 24.7% of patients. Evaluation for elevated ESR and epidural collection can help improve yield and patient selection for biopsy.


Hepatic synthesis of triacylglycerols containing medium-chain fatty acids is dominated by diacylglycerol acyltransferase 1 and efficiently inhibited by etomoxir.
Medium-chain fatty acids (MCFAs) play an increasing role in human nutrition. In the liver, one fraction is used for synthesis of MCFA-containing triacylglycerol (MCFA-TG), and the rest is used for oxidative energy production or ketogenesis. We investigated which enzymes catalyse the synthesis of MCFA-TG and how inhibition of MCFA-TG synthesis or fatty acid (FA) oxidation influences the metabolic fate of the MCFAs. FA metabolism was followed by time-resolved tracing of alkyne-labelled FAs in freshly isolated mouse hepatocytes. Quantitative data were obtained by mass spectrometry of several hundred labelled lipid species. Wild-type hepatocytes and cells from diacylglycerol acyltransferase (DGAT)1-/- mice were treated with inhibitors against DGAT1, DGAT2, or FA β-oxidation. Inhibition or deletion of DGAT1 resulted in a reduction of MCFA-TG synthesis by 70%, while long-chain (LC)FA-TG synthesis was reduced by 20%. In contrast, DGAT2 inhibition increased MCFA-TG formation by 50%, while LCFA-TG synthesis was reduced by 5-25%. Inhibition of β-oxidation by the specific inhibitor teglicar strongly increased MCFA-TG synthesis. In contrast, the widely used β-oxidation inhibitor etomoxir blocked MCFA-TG synthesis, phenocopying DGAT1 inhibition. DGAT1 is the major enzyme for hepatic MCFA-TG synthesis. Its loss can only partially be compensated by DGAT2. Specific inhibition of β-oxidation leads to a compensatory increase in MCFA-TG synthesis, whereas etomoxir blocks both β-oxidation and MCFA-TG synthesis, indicating a strong off-target effect on DGAT1.


Growth of a grazing phytoplanktivorous fish and growth enhancement of the grazed alga.
The blue tilapia, Tilapia aurea, consumes the green alga, Ankistrodesmus falcatus, with some algal cells passing through the gut in viable condition. Grazing fish grew and the grazed algal populations had enhanced densities compared to ungrazed populations. Algal growth enhancement was not produced by either bacteria or fish excreting an algal limiting nutrient. Possible ingestion and digestion of bacterial cells by the fish might release nutrients to the algal cells that were otherwise unavailable. Blue tilapia may be affecting algal communities in ways other than by passive size selection. Enhancement of algal growth by the blue tilapia could have important implications for understanding the phytoplankton dynamics in waters containing blue tilapia.


[Nasal Highflow - Treatment option in severe hypoxaemia?].
Nasal High Flow (NHF) provides a warmed and humidified air stream. In pediatrics, this method is already in widespread use and is increasingly replacing the CPAP. New studies show success in treating adults. Currently the acute hypoxemic insufficiency cause of pneumoniae is a secured area of use. NHF is not inferior in comparison to other oxygen delivery systems. At this juncture effectiveness of this easy to use method is not clear. Preliminary reports describe an improvement in ventilation by the NHF. Here, a wash-out of the airways and improved alveolar ventilation seem to be the main operating principles.


Hydrogen Bonding in Platinum Indolylphosphine Polyfluorido and Fluorido Complexes.
The reaction of the Pt complexes cis-[Pt(CH3 )(Ar){Ph2 P(Ind)}2 ] (Ind=2-(3-methyl)indolyl, Ar=4-tBuC6 H4 (1 a), 4-CH3 C6 H4 (1 b), Ph (1 c), 4-FC6 H4 (1 d), 4-ClC6 H4 (1 e), 4-CF3 C6 H4 (1 f)) with HF afforded the polyfluorido complexes trans-[Pt(F(HF)2 )(Ar){Ph2 P(Ind)}2 ] 2 a-f, which can be converted into the fluoride derivatives trans-[Pt(F)(Ar){Ph2 P(Ind)}2 ] (3 a-f) by treatment with CsF. The compounds 2 a-f and 3 a-f were characterised thoroughly by multinuclear NMR spectroscopy. The data reveal hydrogen bonding of the fluorido ligand with HF molecules and the indolylphosphine ligand. Polyfluorido complexes 2 a-f show larger |1 J(F,Pt)|, but lower 1 J(H,F) coupling constants when compared to the fluorido complexes 3 a-f. Decreasing 1 J(P,Pt) coupling constants in 2 a-f and 3 a-f suggest a cis influence of the aryl ligands in the following order: 4-tBuC6 H4 (a) ≈4-CH3 C6 H4 (b)<Ph (c)≪4-FC6 H4 (d)<4-ClC6 H4 (e)<4-CF3 C6 H4 (f). In addition, the larger cis influence of aryl ligands bearing electron-withdrawing groups in the para position correlates with decreasing magnitudes of |1 J(F,Pt)| coupling constants. The interpretation of the experimental data was supported by quantum-chemical DFT calculations.


The effect of pH on the oxidation of bovine serum albumin by hypervalent myoglobin species.
Bovine serum albumin (BSA) was used as a probe for the oxidation of proteins by hypervalent myoglobin species in solutions with pH from 5.3 to 7.7. The reaction between perferrylmyoglobin, *MbFe(IV)=O, and BSA was studied by activating metmyoglobin with equimolar amounts of hydrogen peroxide in the presence of BSA. A minor pH dependence was observed as judged from the formation of BSA-centered radicals, which were monitored at room temperature by electron spin resonance spectroscopy, and the formation of dityrosine. The reaction between ferrylmyoglobin, MbFe(IV)=O, and BSA was pH-dependent. BSA-centered radicals and dityrosine were formed in low levels at neutral pH and increased at low pH to the same levels as observed in the reaction of *MbFe(IV)=O with BSA. The present results demonstrate that protein-centered radicals can be formed from the non-radical MbFe(IV)=O under mildly acidic conditions, and this should be taken into account when considering oxidation in cellular compartments of low pH and in meat-related products.


Do calcium and calmodulin trigger maturation in amphibian oocytes?
In contrast to earlier reports (J. L. Maller and E. G. Krebs, 1980, Curr. Top. Cell. Regul. 16, 271-311; M. Moreau, J. P. Vilian, and P. Guerrier, 1980, Dev. Biol. 78, 201-214; W. J. Wasserman and L. D. Smith, 1981, J. Cell Biol. 89, 389-394; D. Huchon, R. Ozon, E. H. Fischer, and J. G. Demaille, 1981, Mol. Cell. Endocrinol. 22, 211-222) calmodulin preparations isolated from Xenopus laevis ovaries or obtained commercially rarely induced maturation upon microinjection into individual oocytes. Calmodulin injections did result in significant cases of maturation when oocytes were first pretreated (primed) with calcium-free (EGTA) OR-2 and then injected in regular OR-2 medium. However, under these conditions the injected buffer solution alone was sometimes found to induce maturation. Under more optimal priming conditions, cases were found where as high as 100% of the oocytes matured simply by returning them to regular OR-2 medium. To determine which divalent cations could be involved in the priming effects of EGTA pretreatment we repeated the earlier ionophore work of W. J. Wasserman and Y. Masui (1975, J. Exp. Zool. 193, 369-375), looking not just at calcium and magnesium but other divalent cations as well. Several divalent cations (10 mM) were found to induce germinal vesicle breakdown with the following tentative order of efficacy, Co2+ greater than or equal to Zn2+ greater than or equal to Mn2+ greater than Ca2+ greater than Mg2+ greater than Ba2+, regardless of whether or not ionophore A23187 was present. These results, along with other reports in the literature, are discussed with respect to the theory that a rise in free calcium and calmodulin is involved in triggering oocyte maturation; we conclude that neither is involved.


Evidence on the discrimination of quinoa grains with a combination of FT-MIR and FT-NIR spectroscopy.
Quinoa is considered as a valuable re-emergent crop due to its nutritional composition. In this study, five quinoa grains from different geographical origin (Real, CHEN 252, Regalona, BO25 and UDc9) were discriminated using a combination of FT-MIR and FT-NIR spectra as input for principal component analysis (PCA), cluster analysis (CA) and soft independent modelling class analogy (SIMCA). The results obtained from PCA and CA show a great power of discrimination, with an average silhouette width value of 0.96. Moreover, SIMCA showed an error rate and accuracy values of 0 and 1 respectively with only 4% misclassified samples. A relationship between each principal component and the most important variables for the discrimination were mainly due to vibrations of several oleofins groups (C-H, C-H2, C-H3), alkene group (-CH=CH-), hydroxyl group (O-H) and Amides I and II vibrational modes.


Effect of biologically active plants used as netst material and the derived benefit to starling nestlings.
The European starling Sturnus vulgaris preferentially incorporates fresh sprigs of particular plant species for use as nesting material. Chemicals found in these plants may act to reduce pathogen and ectoparasite populations normally found in nest environments. The present experiments were performed to test this Nest Protection Hypothesis. In the fild, we experimentally determined that wild carrot Daucus carota, a plant species preferred as nest material, effectively reduced the number of hematophagous mites found within nests relative to control nests without green vegetation. Chicks from nests containing wild carrot had higher levels of blood hemoglobin than chicks from control nests. However, there were no differences in weight or feather development. In the laboratory, we found that wild carrot and fleabane, Erigeron philadelphicus, (also preferred by starlings as nest material) substantially reduced the emergence of feeding instars of mites, while garlic mustard, Alliaria officinalis, (commonly available but not preferred) had little effect on the emergence of mites. We infer that preferred plant material may act to inhibit feeding or otherwise delay reproduction of mites, thereby reducing risk of anemia to developing nestlings.


Muscarinic antagonists with multiple stereocenters: Synthesis, affinity profile and functional activity of isomeric 1-methyl-2-(2,2-alkylaryl-1,3-oxathiolan-5-yl)pyrrolidine sulfoxide derivatives.
Completing a long-lasting research on 1,3-oxathiolane muscarinic ligands, we have synthesized a set of isomeric 1-methyl-2-(2,2-alkylaryl-1,3-oxathiolan-5-yl)pyrrolidine 3-sulfoxide derivatives, containing three or four stereogenic centers. In general the compounds are very potent antagonists even if, unlike the corresponding agonists, they show modest subtype selectivity.


Identification of individual amino acids in platelet-derived growth factor that contribute to the specificity towards the beta-type receptor.
Platelet-derived growth factor constitutes a family of three isoforms (PDGF-AA, -AB, and -BB) composed of two homologous polypeptide chains (A and B). These isoforms interact with two types of receptors termed alpha or beta. Whereas PDGF-AA binds only to the alpha-receptor, PDGF-BB binds and activates both receptors with high affinity. To map regions that are specific for the beta-receptor, we introduced mutations into PDGF-AA located in previously identified epitopes [1991, Biochemistry 30, 3303-3309]. A single amino acid exchange in domain II of PDGF-AA (Ala67----Arg) was sufficient to bring about a reduced but significant activation of the beta-receptor. In domain I the exchange of residues Pro26----Arg together with Ser28----Asn switched the specificity towards the beta-receptor. These data indicate that parts of the exposed domains are indeed involved in receptor binding. Since these single mutations lead to mutant proteins which are about 100-fold less active than PDGF-BB, it is suggested that other amino acid residues also participate in the binding to the receptors.


Larvicidal efficacy of five cucurbitaceous plant leaf extracts against mosquito species.
Larvicidal activity of crude hexane, ethyl acetate, petroleum ether, acetone, and methanol extracts of the leaf of five species of cucurbitaceous plants, Citrullus colocynthis, Coccinia indica, Cucumis sativus, Momordica charantia, and Trichosanthes anguina, were tested against the early fourth instar larvae of Aedes aegypti L. and Culex quinquefasciatus (Say) (Diptera: Culicidae). The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in petroleum ether extract of C. colocynthis, methanol extracts of C. indica, C. sativus, M. charantia, and acetone extract of T. anguina against the larvae of A. aegypti (LC50=74.57, 309.46, 492.73, 199.14, and 554.20 ppm) and against C. quinquefasciatus (LC50=88.24, 377.69, 623.80, 207.61, and 842.34 ppm), respectively. The petroleum ether extract of C. colocynthis and methanol extract of M. charantia were more effective than the other extracts. This is an ideal eco-friendly approach for the control of the dengue vector, A. aegypti, and the lymphatic filariasis vector, C. quinquefasciatus.


Association of sleep duration with underweight and obesity in older adults: A cross-sectional study of the Chinese longitudinal healthy longevity survey.
Underweight and obesity are two of the main health concerns as they are associated with adverse health outcomes and nutrition disorders. This study determined the association of sleep duration with both underweight and obesity in Chinese older adults. This cross-sectional secondary data analysis was conducted using data from the 2018 wave of the Chinese Longitudinal Healthy Longevity Survey. There were 13 917 participants aged 65 and above in the analysis. Body mass index (BMI) was categorized into underweight, normal weight and overweight, and obesity. Self-reported sleep duration was divided into short, normal, and long sleep duration. Logistic regression models were used to examine the association of sleep duration with underweight and obesity. The prevalence of underweight was 16.7% and the prevalence of obesity was 8.0% in Chinese older adults aged 65 and older. Compared to the normal sleep duration group, the short sleep duration group had a higher incidence of underweight (AOR = 1.29, 95% CI: 1.01-1.65), and the long sleep duration group showed a higher incidence of obesity (AOR = 1.57, 95% CI: 1.07-2.30) in older men. A greater incidence of underweight was found in the short sleep duration group (AOR = 1.19, 95% CI: 1.02-1.40) in old-old adults after adjusting for covariates. Short sleep duration is associated with being underweight and long sleep duration is linked with obesity in older men. Short sleep duration is also a risk factor for the underweight of old-old adults.


[Study of an homogeneous population of obeses people and of its regression of weight under hypocaloric diet in hospital. II. - Weight loss (author's transl)].
A study of 79 obese patients who were hospitalized and treated with a low calorie diet demonstrated that the weight loss occurred in a univocal manner, according to a semilogarithmic law. It was not influenced, as far as initial results were concerned, by the age, sex, or morphotype of the patient, or whether weight gain was recent or of long-standing. Factors affecting results, however, are the existence of functional signs, the amount of natriuresis during the first few days, and the quantity of alcohol ingested regularly. Short term results, one month after leaving hospital, appear to be moderate only. A diet supplying 800 calories causes weight loss at the expense of body fat. Individual variations in minimal calorie requirements during the treatment can be used as a basis for prescribing a personalized supplementary diet.


Acrylic resin stents for marsupialization.
Marsupialization is a surgical technique that may be preferred to enucleation in treatment of benign cysts. The acrylic resin obturator maintains the surgical opening during healing and assures success of the surgery. Two basic techniques to construct a stable and retentive obturator prosthesis were described.


Systems and synthetic metabolic engineering for amino acid production - the heartbeat of industrial strain development.
With a world market of more than four million tons per year, l-amino acids are among the most important products in industrial biotechnology. The recent years have seen a tremendous progress in the development of tailor-made strains for such products, intensively driven from systems metabolic engineering, which upgrades strain engineering into a concept of optimization on a global scale. This concept seems especially valuable for efficient amino acid production, demanding for a global modification of pathway fluxes - a challenge with regard to the high complexity of the underlying metabolism, superimposed by various layers of metabolic and transcriptional control.


Action of 2-acetamido-2-deoxy-beta-D-hexosidase from Turbo cornutus on periodate-oxidized and Smith-degraded, blood-group HLeb and Lea substances from human, ovarian-cyst fluids.
Immunochemical and chemical studies were used to monitor and evaluate the structural changes produced by an enzyme from Turbo cornutus in periodate-oxidized and Smith-degraded, human blood-group substances from ovarian cysts. After the first step of periodate oxidation and Smith degradation, two blood-group substances, JS (HLeb and N-1 (Lea), were precipitated by mouse-myeloma S117 serum, specific for terminal, nonreducing, beta-D-linked 2-acetamido-2-deoxy-D-glucosyl groups, but not by type XIV antipneumococcal horse serum specific for terminal, nonreducing, beta-D-linked D-galactosyl groups. An exoglycosidase, 2-acetamido-2-deoxy-beta-D-hexosidase (beta-N-acetylhexosaminidase) from Turbo cornutus, split off 2-acetamido-2-deoxy-D-glucose amounting to 22.5 and 20.4% of the total weight of JS and N-1 blood-group substances, respectively. After enzymic digestion, both blood-group substances precipitated with type XIV serum, and did not precipitate with S117 serum. The findings are in agreement with the structure propsed for the water-soluble, blood-group substances [Lloyd and Kabat, Proc. Natl. Acad. Sci. U.S.A., 61 (1968) 1477]. Specific enzymes can be of value in structural studies when used in conjunction with sequential periodate oxidation and Smith degradation.


Tsc1 represses parvalbumin expression and fast-spiking properties in somatostatin lineage cortical interneurons.
Medial ganglionic eminence (MGE)-derived somatostatin (SST)+ and parvalbumin (PV)+ cortical interneurons (CINs), have characteristic molecular, anatomical and physiological properties. However, mechanisms regulating their diversity remain poorly understood. Here, we show that conditional loss of the Tuberous Sclerosis Complex (TSC) gene, Tsc1, which inhibits the mammalian target of rapamycin (MTOR), causes a subset of SST+ CINs, to express PV and adopt fast-spiking (FS) properties, characteristic of PV+ CINs. Milder intermediate phenotypes also occur when only one allele of Tsc1 is deleted. Notably, treatment of adult mice with rapamycin, which inhibits MTOR, reverses the phenotypes. These data reveal novel functions of MTOR signaling in regulating PV expression and FS properties, which may contribute to TSC neuropsychiatric symptoms. Moreover, they suggest that CINs can exhibit properties intermediate between those classically associated with PV+ or SST+ CINs, which may be dynamically regulated by the MTOR signaling.


Influence of the physical properties and handling of silica gel modified carbon paste electrodes on the phase transfer of solved Cu(II) ions.
Carbon paste electrodes (CPE) modified with natural minerals might be utilised to mimic adsorption/desorption reactions at natural interfaces. As a step towards this aim, several properties and application conditions of CPEs, modified with silica gel, were tested for their influence on phase transfer reactions of solved Cu(II)-tetrammine complex ions. The peak current recorded during the anodic stripping of adsorbed Cu(II) species increased with increasing diameter of the circular electrode surface. A silica gel content of 29 mass percent yielded the highest peak current at otherwise constant exposition conditions. A substitution of graphite by glassy carbon did not enhance the electrode sensitivity. The peak current increased with increasing exposition time (up to 20 min) and with decreasing size of the modifier particles. Typically between 12% and 22% of the solved Cu ions were adsorbed onto the electrode surface and between 15% and 20% of this Cu fraction was subsequently removed by stripping and electrode regeneration. Despite the incomplete electrode regeneration the mass transfer ratios of the Cu(II) species were relatively constant during several application cycles.


A comparison of calisthenic and cycle exercise training in chronic obstructive pulmonary disease patients: a randomized controlled trial.
To compare the effects of calisthenic and cycle exercises with no exercise in chronic obstructive pulmonary disease patients. Forty-seven participants were allocated to either a cycle or calisthenic exercise or control group. Outcome measures, including Saint George Respiratory Questionnaire, pulmonary functions, cardiopulmonary exercise testing, Fitness Testing, and Hospital Anxiety-Depression, Modified Medical Research Council Dyspnea, Fatigue Severity, Fatigue Impact Scales, were performed before and after the intervention. The change in VE/VCO2 significantly differed (p = 0.01) between two exercise groups. Physical fitness, quality of life, anxiety-depression, dyspnea and fatigue changed significantly in exercise groups, with no between-group differences. There were no significant improvements in control group. Calisthenics are as safe and effective as cycle exercise and could be included in comprehensive treatment programs.


Effect of palonosetron and dexamethasone administration on the prevention of gastrointestinal symptoms in hepatic arterial chemoembolization with epirubicin.
There are several studies on premedication to prevent postembolization syndromes which occurs after transcatheter arterial chemoembolization (TACE), but the medication to be used is still not established. This study aimed to examine the effect of palonosetron and dexamethasone on the prevention of gastrointestinal symptoms induced by TACE. Patients with hepatocellular carcinoma who were treated with TACE with epirubicin were retrospectively evaluated. The complete response rate of antiemetic drugs and incidence and severity of gastrointestinal symptoms were compared between the antiemetic group (AE group), which includes 51 patients prophylactically administered with palonosetron 0.75 mg and dexamethasone 9.9 mg intravenously before TACE on day 1 and dexamethasone 6.6 mg intravenously on days 2 and 3, and control group with 101 patients without antiemetic premedication. Complete response rate in the entire evaluation period was significantly higher in the AE group compared with that in the control group. In the acute phase, the incidence and severity of nausea, vomiting, and anorexia significantly decreased in the AE group, but only anorexia improved in the delay phase. Additionally, postembolization syndromes, such as abdominal pain and fever, were significantly attenuated in the AE group; however, constipation worsened in this group. Premedication of palonosetron and dexamethasone significantly prevents the incidence and reduces the severity of gastrointestinal symptoms especially in the acute phase. Further studies will be needed to determine the most recommended 5-HT3 antagonist or dosage of dexamethasone in establishing the optimal antiemetic regimen.


Therapeutic effect of Pseudomonas aeruginosa phage YH30 on mink hemorrhagic pneumonia.
Hemorrhagic pneumonia caused by Pseudomonas aeruginosa remains one of the most costly infectious diseases among farmed mink and commonly leads to large economic losses during mink production. The objective of this study was to investigate the potential of using phages as a therapy against hemorrhagic pneumonia in mink. A broad-host-range phage from the Podoviridae family, YH30, was isolated using the mink-originating P. aeruginosa (serotype G) D7 strain as a host. The genome of YH30 was 72,192bp (54.92% G+C), contained 86 open reading frames and lacked regions encoding known virulence factors, integration-related proteins or antibiotic resistance determinants. These characteristics make YH30 eligible for use in phage therapy. The results of a curative treatment experiment demonstrated that a single intranasal administration of YH30 was sufficient to cure hemorrhagic pneumonia in mink. The mean colony count of P. aeruginosa in the blood and lung of YH30-protected mink was less than 10(3) CFU/mL (g) within 24h of bacterial challenge and ultimately became undetectable, whereas that in unprotected mink reached more than 10(8) CFU/mL (g). Additionally, YH30 dramatically improved the pathological manifestations of lung injury in mink with hemorrhagic pneumonia. Our work demonstrates the potential of phages to treat P. aeruginosa-caused hemorrhagic pneumonia in mink.


Human chorionic gonadotropin in the treatment of HIV-related Kaposi's sarcoma.
To evaluate the antineoplastic activity of human chorionic gonadotropin (hCG) in the treatment of HIV-related Kaposi's sarcoma (KS), two clinical trials focusing on two different schedules of administration and types of hCG preparation were conducted. In the low-dose group, hCG (Profasi-HP) was administered three times a week intramuscularly at a dose ranging from 4000 to 32,000 IU for 4 months and no objective response was observed among 5 evaluable patients. In the high-dose group, hCG (Gonadotrafon) was given intramuscularly three times a week at a dose ranging from 100,000 to 300,000 IU for 3 months with 1 partial response among 8 evaluable patients. In 6 patients evaluated for HIV viral load, no significant reduction in HIV viraemia was observed. In conclusion, hCG showed very limited activity against KS and no influence on HIV viral load, along with emerging dose-limiting toxicity and high cost of the therapy.


Cancer in rodents: does it tell us about cancer in humans?
Information obtained from animal models (mostly mice and rats) has contributed substantially to the development of treatments for human cancers. However, important interspecies differences have to be taken into account when considering the mechanisms of cancer development and extrapolating the results from mice to humans. Comparative studies of cancer in humans and animal models mostly focus on genetic factors. This review discusses the bio-epidemiological aspects of cancer manifestation in humans and rodents that have been underrepresented in the literature.


Isolation and spectral characteristics of the photochemical reaction center of Rhodopseudomonas viridis.
A method is described for isolation of the Rhodopseudomonas viridis reaction center complex free of altered, 685 nm absorbing pigment. This improved preparation contains two c-type cytochromes in the ratio P-960: cytochrome c-558: cytochrome c-553 of 1:2:2 to 3. The near infrared spectral forms of the reduced preparation are located at 790, 832, 846, and 987 nm at 77 K; the oxidized complex absorbs at 790, 808, 829 and approx. 1310 nm. The 790 nm band is attributed to bacteriophaeophytin b and the other absorbances to bacteriochlorophyll b, The visible absorption bands may be assigned to these pigments and to the cytochromes present and, probably to a carotenoid. The presence of two bacteriochlorophyll b spectral forms in the P+-830 band suggests that exciton interactions occur among pigments in the oxidized, as well as the reduced, reaction center. Changes in the 790 and 544 nm bands upon illumination of the reaction center preparation at low redox potential may be indicative of a role for bacteriophaeophytin b in primary photochemical events.


Alcohol binge drinking decreases brain glucose metabolism and functional connectivity in adolescent rats.
Alcohol misuse represents a serious health concern, especially during adolescence, with approximately 18% of high school students engaging in binge drinking. Despite widespread misuse of alcohol, its effects on how the brain functions is not fully understood. This study utilized a binge drinking model in adolescent rats to examine effects on brain function as measured by brain glucose metabolism (BGluM). Following an injection of [18 FDG] fluro-2-deoxy-D-glucose, rats had voluntary access to either water or various concentrations of ethanol to obtain the following targeted doses: water (no ethanol), low dose ethanol (0.29 ± 0.03 g/kg), moderate dose ethanol (0.98 ± 0.05), and high dose ethanol (2.19 ± 0.23 g/kg). Rats were subsequently scanned using positron emission tomography. All three doses of ethanol were found to decrease BGluM in the restrosplenial cortex, visual cortex, jaw region of the somatosensory cortex, and cerebellum. For both the LD and MD ethanol dose, decreased BGluM was seen in the superior colliculi. The MD ethanol dose also decreased BGluM in the subiculum, frontal association area, as well as the primary motor cortex. Lastly, the HD ethanol dose decreased BGluM in the hippocampus, thalamus, raphe nucleus, inferior colliculus, and the primary motor cortex. Similar decreases in the hippocampus were also seen in the LD group. Taken together, these results highlight the negative consequences of acute binge drinking on BGluM in many regions of the brain involved in sensory, motor, and cognitive processes. Future studies are needed to assess the long-term effects of alcohol binge drinking on brain function as well as its cessation.


Evaluating evidence-based medicine skills during a performance-based examination.
To measure students' competencies in evidence-based medicine (EBM) skills [clinical decision making using evidence from published literature (content) and in transmitting clinical information to patients (communication)] within the context of a performance-based examination (PBE). In 2002-03, under the direction of a Performance-Based Examination Oversight Committee, 16 EBM queries were developed for a pair of third-year PBEs. At the last station of the PBE, the standardized patient (SP) for that station asked a clinical EBM question relating to their "disease process." Students were asked to develop an appropriate clinical question, perform a Medline search for appropriate articles, critically appraise a complete selected article, reach a conclusion to their question, and transmit the information to the SP. Each student's clinical question, search terms, selected articles, and rationale were evaluated by faculty question-writers, clinical librarians, and the EBM course director using a five-point Likert scale, with 1 being inadequate performance and 5 being superior performance. The SP evaluated the communication skills using a checklist. Students' performances were very good, with means of 3.7 to 4.0 in each area. Agreement between the course director and station developers was good. Seventy-five percent of the students performed adequate Medline searches. Students averaged over 93% on the performance of four communication skills. The evaluation of EBM skills can be carried out during a performance-based examination. Results can assist in developing students' skills and directing curricular efforts.


Ursodeoxycholate protects against ethanol-induced liver mitochondrial injury.
The purpose of this work was to examine whether ursodeoxycholate (UDC), a hydrophilic bile salt, could reduce mitochondrial liver injury from chronic ethanol consumption in rats. Animals were pair-fed liquid diets containing 36% of calories as ethanol or isocaloric carbohydrates. They were randomly assigned into 4 groups of 7 rats each and received a specific treatment for 5 weeks: control diet, ethanol diet, control diet + UDC, and ethanol diet + UDC. Respiratory rates of isolated liver mitochondria were measured using a Clark oxygen electrode with sodium succinate as substrate. Mitochondria from rats chronically fed ethanol demonstrated an impaired ability to produce energy. At the fatty liver stage, the ADP-stimulated respiration (V3) was depressed by 33%, the respiratory control ratio (RC) by 25% and the P/O ratio by 15%. In ethanol-fed rats supplemented with UDC, both the rate and efficiency of ATP synthesis via the oxidative phosphorylation were improved: V3 was increased by 35%, P/O by 8%. All the respiratory parameters were similar in control group and control + UDC group. On the other hand, the number and size of mitochondria were assessed by electron microscopy and computer-assisted quantitative analysis. The number of mitochondria from ethanol-treated rats was decreased by 29%, and they were enlarged by 74%. Both parameters were normalized to control values by UDC treatment. These studies demonstrate that UDC has a protective effect against ethanol-induced mitochondrial injury by improving ATP synthesis and preserving liver mitochondrial morphology. These UDC positive effects may contribute to the observed decrease in fat accumulation and may delay the progression of alcoholic injury to more advanced stages.


Economics of baculovirus-insect cell production systems.
Commercial production of pharmaceutical proteins in baculovirus - insect cell systems is already a reality, and has therefore not been discussed in detail here. Cost-efficacy will depend on the productivity of the protein in culture, the dose, and the quantities required. According to the model described here, cost-effective production of baculoviruses for use in agriculture should also be feasible, assuming the commercial availability of a low-cost medium, together with a baculovirus with high productivity in cell culture, which is effective at a field application rate of 10(12) PIB ha(-1) or lower. All of these criteria appear to be achievable, given fairly modest advances over currently available technology. Given the relatively high fixed costs associated with production of baculoviruses on an agricultural scale in bioreactors however, profitability will depend on the scale of production. A substantial market opportunity (perhaps in the order of 1 million hectares) would be necessary in order to exploit the economies of scale achievable with baculovirus -insect cell production systems.


Pregabalin drug interaction studies: lack of effect on the pharmacokinetics of carbamazepine, phenytoin, lamotrigine, and valproate in patients with partial epilepsy.
Pregabalin (PGB) is an alpha2-delta ligand with demonstrated efficacy in epilepsy, neuropathic pain, and anxiety disorders. PGB is highly efficacious as adjunctive therapy in patients with refractory partial seizures. Given its efficacy as adjunctive therapy, the potential for interaction of PGB with other antiepileptic drugs (AEDs) was assessed in patients with partial epilepsy in open-label, multiple-dose studies. Patients received PGB, 600 mg/day (200 mg q8h) for 7 days, in combination with their individualized maintenance monotherapy with valproate (VPA), phenytoin (PHT), lamotrigine (LTG), or carbamazepine (CBZ). Trough steady-state concentrations of CBZ (and its epoxide metabolite), PHT, LTG, and VPA were unaffected by concomitant PGB administration. Likewise, PGB steady-state pharmacokinetic parameter values were similar among patients receiving CBZ, PHT, LTG, or VPA and, in general, were similar to those observed historically in healthy subjects receiving PGB alone. The PGB-AED combinations were generally well tolerated. PGB may be added to VPA, LTG, PHT, or CBZ therapy without concern for pharmacokinetic drug-drug interactions.


Sleep/waketime preference and delayed diurnal eating rhythms are associated through light exposure timing and modified by sleep efficiency.
Delayed eating rhythms, relative to the sleep/wake period, commonly manifest as a lack of hunger in the morning (morning anorexia) and elevated hunger in the late evening (evening hyperphagia). These intake patterns are associated with adverse mental and physical health outcomes. We aimed to evaluate whether the timing of light exposure, an important environmental signal for circadian synchronization, explains the link between sleep/waketime preferences and delayed diurnal appetite. We also aimed to test whether disruptions in sleep quality, reflecting suboptimal circadian synchronization, identify individuals for whom sleep/waketime preference is associated with delayed diurnal appetite. Participants (N = 150) completed a measure of their sleep/waketime preferences and wore a device to capture their sleep efficiency and naturalistic light exposure for 48 consecutive hours. The timing of light exposure mediated the link between sleep/waketime preferences and evening hyperphagia, but not morning anorexia, such that a later peak in light exposure mitigated some of the risk for evening hyperphagia that was associated with later sleep/waketime preferences. Sleep efficiency moderated the association between sleep/waketime preference and morning anorexia, but not evening hyperphagia. Earlier sleep/waketime preference was associated with less morning anorexia among individuals with high sleep efficiency, but morning anorexia was consistently elevated among individuals with poor sleep efficiency. These results on the relation between sleep/waketime preference and two aspects of delayed diurnal appetite suggest that morning anorexia depends on sleep efficiency and evening hyperphagia may be influenced by the timing of daily light exposure. Future research should assess over longer periods, covering weekdays and weekends, and incorporate momentary reports of meals/snacks and appetite.


Species differences in the hepatotoxicity of paracetamol are due to differences in the rate of conversion to its cytotoxic metabolite.
The cytotoxicity of paracetamol and of its putative toxic metabolite, N-acetyl-p-benzo-quinoneimine (NABQI) have been investigated in hepatocytes from hamster, mouse, rat and human liver. Whereas paracetamol readily caused cell blebbing and a loss of viability in hepatocytes from mouse and hamster, human and rat hepatocytes were much more resistant to these effects. In marked contrast, there were no significant differences in the sensitivity of the cells from any species to the toxic effects of NABQI. Glutathione depletion by NABQI and paracetamol correlated very well with the toxic effects of these compounds. It is concluded that species differences in sensitivity to the hepatotoxicity of paracetamol are due almost entirely to differences in the rate of formation of NABQI, and not to any intrinsic differences in sensitivity or in any difference in the fate of NABQI once formed. Further, man appears to be relatively resistant to the hepatotoxic effects of paracetamol, and the results in hepatocytes were confirmed by both in vitro and in vivo analyses.


Considering patient values and treatment preferences enhances patient involvement in rectal cancer treatment decision making.
The shared decision making (SDM) model states that patients' values and preferences should be clarified to choose a strategy that best fits the patient. This study aimed to assess whether values and preferences of rectal cancer patients are voiced and considered in deciding about preoperative radiotherapy (PRT), and whether this makes patients feel more involved in treatment decision making. Pre-treatment consultations of radiation oncologists and patients eligible for PRT were audiotaped (N=90). Tapes were transcribed and coded to identify patients' values and treatment preferences. Patients filled in a post-consultation questionnaire on their perceived involvement in decision making (N=60). Patients' values were voiced for 62/611 of benefits/harms addressed (10%), in 38/90 consultations (42%; maximum 4 values per consultation), and most often related to major long-term treatment outcomes. Patients' treatment preferences were discussed in 20/90 consultations (22%). In 16/90 consultations (18%), the oncologists explicitly indicated to consider patients' values or preferences. Patients perceived a significantly more active role in decision making if their values or preferences had been voiced or considered. Patients' values and treatment preferences are voiced or considered in a minority of consultations. If they are, this increases patients' perceived involvement in the decision making process.


Gynogenic plant regeneration from unpollinated flower explants of Eragrostis tef (Zuccagni) Trotter.
Tef [Eragrostis tef (Zucc.) Trotter] is the most important cereal in Ethiopia. In its wild relative E. mexicana, regeneration of six green plants resulted from culture of 121 non-pollinated immature pistils. In the allotetraploid crop species tef, however, only callus and root formation was obtained by this method. By contrast, immature spikelets and panicle segments of E. tef proved amenable to gynogenic plant regeneration. Upon step-wise optimization of the protocol, efficient plant formation was achieved in all three cultivars tested. In cv. DZ-01-196, culture of 1305 immature spikelets resulted in formation of 159 green plants. Flow cytometric analysis revealed (di)haploid, triploid, tetraploid and octoploid regenerants, from which the vast majority was tetraploid. Tef-breeding programs will likely benefit substantially from efficient generation of true-breeding plants.


The effect of eye closure on the post-lens tear film thickness during silicone hydrogel contact lens wear.
The purpose of this study was to investigate the effect of eye closure on the thickness on the post-lens tear film (POLTF) during silicone hydrogel contact lens wear. Ten subjects (mean age, 30.2 +/- 8.6 years; seven males) wore balafilcon A silicone hydrogel contact lenses in both eyes during the experimental procedure (power = -2.00 D, base curve = 8.6 mm). Previously described interference techniques, based on oscillations in reflectance spectra, were used to measure the post-lens tear film. Reflectance spectra (562-1030 nm) from the front of the eye wearing a contact lens were measured at normal incidence where the thickness of the POLTF was derived from the "frequency" of the oscillations. Six baseline measures of POLTF thickness were taken, followed by eye closure in the supine position for 30 minutes. Post-lens tear film thickness measures were taken after 5 and 15 minutes of eye closure. After 30 minutes of eye closure, 1 measure of POLTF was taken every minute for an additional 15 minutes. The average baseline POLTF thickness was 2.00 +/- 0.30 microm (median, 2.00) for the 10 subjects. The average POLTF thickness decreased to 1.58 +/- 0.33 and 1.20 +/- 0.23 microm after 5 and 15 minutes of eye closure, respectively (repeated measures ANOVA, F = 42.18, P < 0.0001). Only 5 of 60 POLTF thickness estimates were obtained for the first 6 minutes following completion of 30 minutes of eye closure, indicating that the POLTF may be too thin (i.e., <1 microm) to measure using this method. However, there was a significant increase in POLTF thickness for 15 minutes after prolonged eye closure (mixed modeling regression, thickness = 0.85 + 0.07 x time after eye opening; P < 0.0001). The POLTF thickness is rapidly reduced by eye closure during contact lens wear. After 30 minutes of eye closure, the POLTF thickness may be reduced to values less than 1 microm for the first several minutes after opening the eyes.


Peroxynitrite affects the cumulus cell defense of metaphase II mouse oocytes leading to disruption of the spindle structure in vitro.
To demonstrate the effects of peroxynitrite (ONOO(-)) on metaphase II mouse oocyte spindle structure and chromosomal alignment in presence and absence of cumulus cells. Experimental study. University-based research laboratory. Metaphase II mouse oocytes (n = 440). Metaphase II mouse oocytes, with and without cumulus cells, were exposed to ONOO(-), nitrite/nitrate, the final product of ONOO(-), and nontreated controls for 15 minutes. Oocytes were fixed and subjected to indirect immunofluorescence for detecting changes in the spindle and chromosomal alignment. Viability staining in exposed oocytes with and without cumulus cells was performed using the trypan blue dye exclusion method and compared with controls. Scoring the alterations in spindle and chromosomal alignment using immunofluorescent and confocal microscopy based on a previously validated system. Most oocytes had poor scores for the spindle and chromosomal alignment with exposure to ONOO(-) in a dose-dependent manner compared with controls. Trypan blue staining revealed that most of the cumulus cells failed to survive treatment with ONOO(-) compared with controls. ONOO(-) affects the viability of cumulus cells and the oocyte spindle structure in a dose-dependent manner. Collectively, these effects compromise oocyte quality, which may lead to female infertility.


Low-density lipoprotein cholesterol, statin therapy, and cerebral microbleeds: The CIRCLE study.
Current evidence suggests a potential association between cerebral microbleeds (CMBs), low-density lipoprotein cholesterol (LDL-C) levels, and statin use, but the exact relationship remains unclear. This study aims to prospectively examine these relationships in a stroke-free population. From January 2010 to January 2020, we enrolled stroke-free individuals with at least one cerebral small vessel disease imaging marker from the CIRCLE study (ClinicalTrials.gov ID: NCT03542734). Participants underwent baseline and 1-year follow-up susceptibility-weighted imaging (SWI), and baseline LDL-C testing. New CMBs were categorized as strictly lobar and deep CMBs based on location. A total of 209 individuals were included. Baseline serum LDL-C levels were divided into quartiles: Q1 (≤1.76 mmol/L), Q2 (1.77-2.36 mmol/L), Q3 (2.37-2.93 mmol/L), and Q4 (>2.93 mmol/L). The incidence of new deep CMBs was 30.0%, 11.1%, 10.9%, 8.2% in Q1, Q2, Q3, Q4, respectively. Multivariate logistic model revealed that only LDL-C in Q1 was associated with increased incidence of new deep CMBs (OR = 4.256; 95% CI: 1.156-15.666; p = 0.029). In a subset of 169 participants without prior statin use, the use of atorvastatin was associated with reduced occurrence of new deep CMBs (OR = 0.181; 95% CI: 0.035-0.928; p = 0.040), while it was not found with rosuvastatin (OR = 0.808; 95% CI: 0.174-3.741; p = 0.785). While lower LDL-C levels were associated with higher CMB development, statin therapy did not increase the risk of new CMBs. Atorvastatin even demonstrated a protective effect.


Comparison of two fluorescence immunoassay methods for the detection of endocrine disrupting chemicals in water.
We describe two fluorescence immunoassays capable of detecting endocrine disrupting compounds in waste water. The first fluorescence method is a heterogeneous assay using total internal reflection fluorescence (TIRF) detection. The second method is a homogeneous assay that utilizes energy transfer (ETIA). Both fluorescence immunoassays are compared with respect to detection principle and ability to quantify the model analytes estrone, estradiol, and ethinylestradiol in a complex matrix regarding recovery rates and limits of detection. Calibrations were performed for the three analytes using both fluorescence methods. Limits of detection between 0.01 and 0.85 microg/l are achieved. In addition, measurements in synthetic waste water spiked with the analytes were performed. Both immunoassays allow the detection in waste water with recovery rates in the range of 70-112%.


Pharmacokinetics and toxicity evaluation following oral exposure to bisphenol F.
Bisphenol F is a substitute material for bisphenol A and is widely used in household products as a raw material for polycarbonate resin, epoxy resin, and plastic reinforcement. It is known to be mainly used in food containers, thermal paper for receipts, and coatings for water pipes. In some countries, bisphenol F has been detected in drinking water and human urine samples. However, due to the lack of safety evaluation data on bisphenol F, it is difficult to establish appropriate guidelines for the proper use of the substance, and social anxiety is increasing accordingly. This study investigated the use, exposure route, and distribution flow of bisphenol F, a household chemical. To determine the no-observed-adverse-effect level (NOAEL) and target organ of bisphenol F after exposure, a single-dose oral toxicity, dose-range finding (28 day oral), repeated dose toxicity (90 day oral), and genotoxicity (reverse mutation, chromosomal abnormality, in vivo micronucleus test) tests were performed. The pharmacokinetic profile was also obtained. The test results are as follows: in the pharmacokinetic study, it was confirmed that single oral exposure to BPF resulted in systemic exposure; in single oral dose toxicity test, the approximate lethal dose was found to be 4000 mg/kg and confusion and convulsion was shown in the test animals; NOAEL was determined to be 2 mg/kg/day for male and 5 mg/kg/day for female, and the no-observed-effect level (NOEL) was determined to be 2 mg/kg/day for males and 1 mg/kg/day for females, and the target organ was the small intestine; genotoxicity tests confirmed that BPF does not induce genotoxicity.


Effects of rosuvastatin and atorvastatin on nonsustained ventricular tachycardia in patients with ST-elevation myocardial infarction: a retrospective analysis.
Early and intensive atorvastatin treatment can decrease nonsustained ventricular tachycardia (nsVT) in patients with ST-segment elevation myocardial infarction (STEMI). The objective of this study was to compare the effects of hydrophilic rosuvastatin and lipophilic atorvastatin on nsVT in STEMI patients treated with primary percutaneous coronary intervention (PCI). The data from a cohort of patients undergoing primary PCI at Jinhua Municipal Central Hospital from January 1, 2013 through June 30, 2016 were analyzed. The patients were divided into the rosuvastatin group and the atorvastatin group based on which kind of statins that they had received. The endpoint of the study was the occurrence of nsVT on either electrocardiogram monitoring or Holter monitoring. A total of 301 patients were enrolled in the study (rosuvastatin group: n = 103; atorvastatin group: n = 198). The baseline and procedural characteristics were similar between the two groups, except that total ischemic time in the rosuvastatin group was markedly longer than that in the atorvastatin group (8 (5-16) h vs. 6 (4-12) h; P = 0.001). The administration of rosuvastatin was significantly associated with lower occurrence of nsVT than that of atorvastatin (9.71 vs. 19.70%; P = 0.026). Multivariable logistic regression analysis suggested that the independent predictors of nsVT included rosuvastatin (odds ratio (OR) 0.397, 95% confidence interval (CI) 0.176-0.894), current smoking (OR 2.307, 95% CI 1.011-5.262), and left ventricular ejection fraction (LVEF) (OR 1.060, 95% CI 1.023-1.098). The effects of rosuvastatin on nsVT might be better than that of atorvastatin in STEMI patients undergoing primary PCI.


Treatment of TNF mediated diseases by selective inhibition of soluble TNF or TNFR1.
The TNF signaling pathway is a valuable target in the therapy of autoimmune diseases, and anti-TNF drugs are successfully used to treat diseases such as rheumatoid arthritis, Crohn's disease and psoriasis. By their ability to interfere with inflammatory processes at multiple levels, these TNF blockers have become invaluable tools to inhibit the inflammation induced damage and allow recovery of the affected tissues. Unfortunately this therapy has some drawbacks, including increased risk of infection and malignancy, and remarkably, the onset of new auto-immune diseases. Some of these effects are caused by the unwanted abrogation of beneficial TNF signaling. More specific targeting of the pathological TNF-induced signaling might lead to broader applicability and improved safety. Specificity might be increased by inhibiting the soluble TNF/TNFR1 axis while leaving the often beneficial transmembrane TNF/TNFR2 signaling untouched. This approach looks promising because it inhibits the pathological effects of TNF and reduces the side effects, and it opens the way for the treatment of other diseases in which TNFR2 inhibition is detrimental. In this review we give an overview of in vivo mouse studies of TNF mediated pathologies demonstrating that the blockade or genetic deletion of sTNF or TNFR1 is preferable over total TNF blockade.


Current efforts towards safe and effective live attenuated vaccines against African swine fever: challenges and prospects.
African swine fever (ASF) is a fatal hemorrhagic disease in domestic pigs and wild boar caused by African swine fever virus (ASFV). Since ASF has been introduced into Europe and Asia, the major pig-raising areas, posing a huge threat to the pork industry worldwide. Currently, prevention and control of ASF are basically dependent on strict biosecurity measures and stamping-out policy once ASF occurs. The major risks of ASF spread are insufficient biosecurity measures and human behaviors. Therefore, a safe and effective vaccine seems to be a reasonable demand for the prevention and control of ASF. Due to the efficacy advantage over other types of vaccines, live attenuated vaccines (LAVs), especially virulence-associated genes deleted vaccines, are likely to be put into emergency and conditional use in restricted areas if ASF is out of control in a country with a huge pig population and pork consumption, like China. However, the safety, efficacy, and genetic stability of current candidate ASF LAVs require comprehensive clinical evaluations prior to country-wide field application. Several critical issues need to be addressed to commercialize an ideal ASF LAV, including a stable cell line for manufacturing vaccines, differentiation of infected from vaccinated animals (DIVA), and cross-protection from different genotypes. A safe and effective DIVA vaccine and an accompanying diagnostic assay will facilitate the prevention, control, and eradication of ASF, which is quite challenging in the near future.


A new SLC12A3 founder mutation (p.Val647Met) in Gitelman's syndrome patients of Roma ancestry.
Gitelman's syndrome (GS) is an autosomal recessive disorder caused by mutations in the SLC12A3 gene. GS is characterized by hypokalaemic metabolic alkalosis, hypomagnesemia and hypocalciuria. Most of the reported patients of Roma ancestry are homozygous for an SLC12A3 intron 9 frameshifting mutation (c.1180+1G>T). Some forms of Bartter's syndrome result from mutations in the CLNCKB gene and clinically overlap with GS. To characterize a second SLC12A3 mutation in Roma patients negative for the intron 9 variant. SLC12A3 and CLNCKB genes were analyzed by next-generation sequencing in two Spanish and Greek gypsy patients who were negative for the intron 9 splicing mutation. Sanger sequencing was performed to confirm the putative mutations in patients and family members. We identified a missense variant (p.Val647Met, c.1939G>A) in both cases, and both were homozygous for Met. This mutation was also found in three additional patients; two homozygous and one heterozygous compound with the intron 9 splicing mutation. This new SLC12A3 mutation seems to be characteristic of gipsy GS patients and was linked to the same haplotype in all cases, supporting a founder origin. All the patients showed biochemical features characteristic of GS. We report a second founder mutation among GS patients of Roma ethnic background. The direct screening of this mutation would facilitate the characterization of patients who are negative for the more common intron 9 +1G>T mutation.


Corrigendum: The silent epidemic of diabetic ketoacidosis at diagnosis of type 1 diabetes in children and adolescents in italy during the covid-19 pandemic in 2020.
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The primary structure of initiation factor IF3 from Bacillus stearothermophilus.
The complete amino acid sequence of the initiation factor IF3 from Bacillus stearothermophilus has been elucidated. This was achieved by splitting the protein with trypsin, Staphylococcus protease or cyanogen bromide. The amino acid sequence was determined by manual Edman degradation, using the DABITC/PITC double-coupling method. The IF3 molecule contains 171 amino acids and has an Mr of 19 677. The sequence was compared to the homologous molecule from Escherichia coli; about 50% of the amino acid residues were found to be identical.


On the genotoxicity of the pesticides Endodan and Kilacar in 6 different test systems.
Two pesticides, the fungicide Endodan (ethylene thiuram monosulphide) and the insecticide-acaricide Kilacar (bis(parachlorophenyl)cyclopropyl methanol), produced or used in the neighbouring countries of Bulgaria and Greece were investigated in a coordinated research programme for their genotoxic effects in a variety of test systems. This included the Ames test, Aspergillus nidulans for mitotic segregation, in vitro human lymphocyte cell cultures for SCE and chromosomal aberrations, in vivo bone marrow cells in hamsters and rats and the dominant lethal test in rats. The genotoxicity of Endodan was found to range from negative to slightly positive in different test systems. At concentrations of 7.5 and 12.0 micrograms/plate together with S9 mix it induced base-pair substitutions in the TA100 strain of Salmonella typhimurium at a rather low level. At a dose of 93 mg/kg b.w. it also caused chromosomal aberrations in acutely treated hamster bone marrow cells. A significant increase of SCE was also found in human lymphocyte cultures at a concentration of 20.0 micrograms/ml. Endodan was found to be negative in A. nidulans for somatic segregation, lymphocyte cultures for chromosomal aberrations and mitotic activity and in rats for dominant lethals and chromosomal aberrations. Kilacar was found to be a weak mutagen in the TA97 strain of S. typhimurium at concentrations of 2.5 and 5.0 micrograms/plate together with S9 mix. At concentrations of 1.0, 1.5 and 2 micrograms/ml Kilacar increased the number of mitotic segregants in A. nidulans by 160%, 220% and 156% respectively over the control. In Syrian hamster bone marrow cells after acute administration at concentrations of 0, 40, 80 and 160 mg/kg, the MI was 5.50, 4.30, 3.10 and 1.30 respectively, and an increase in chromosomal aberrations of about 300% over the control was observed with a concentration of 80 mg/kg. In human lymphocytes no significant changes were observed in either MI or SCE. In the dominant lethal test after chronic treatment of male rats at doses of 5.1, 10.2 and 102.0 mg/kg b.w. no significant mutagenic effect was found although a decrease was shown in the percentage of females with implants mated with treated males in the first week.


Fuzzy-based hunger games search algorithm for global optimization and feature selection using medical data.
Feature selection (FS) is one of the basic data preprocessing steps in data mining and machine learning. It is used to reduce feature size and increase model generalization. In addition to minimizing feature dimensionality, it also enhances classification accuracy and reduces model complexity, which are essential in several applications. Traditional methods for feature selection often fail in the optimal global solution due to the large search space. Many hybrid techniques have been proposed depending on merging several search strategies which have been used individually as a solution to the FS problem. This study proposes a modified hunger games search algorithm (mHGS), for solving optimization and FS problems. The main advantages of the proposed mHGS are to resolve the following drawbacks that have been raised in the original HGS; (1) avoiding the local search, (2) solving the problem of premature convergence, and (3) balancing between the exploitation and exploration phases. The mHGS has been evaluated by using the IEEE Congress on Evolutionary Computation 2020 (CEC'20) for optimization test and ten medical and chemical datasets. The data have dimensions up to 20000 features or more. The results of the proposed algorithm have been compared to a variety of well-known optimization methods, including improved multi-operator differential evolution algorithm (IMODE), gravitational search algorithm, grey wolf optimization, Harris Hawks optimization, whale optimization algorithm, slime mould algorithm and hunger search games search. The experimental results suggest that the proposed mHGS can generate effective search results without increasing the computational cost and improving the convergence speed. It has also improved the SVM classification performance.


BAC manipulations for making BAC transgene arrays.
Chromosome tagging using lac or tet operator repeats for in vivo visualization of chromosome dynamics has now become a standard methodology used in a range of organisms. One variation of this approach has been to build transgene arrays creating artificial chromosome blocks to study various aspects of chromatin structure, transcription, replication, or DNA repair. Previously, plasmid transgenes with or without subsequent gene amplification have been used to build these arrays. However, plasmid arrays typically show heterochromatic properties, while gene amplification typically results in chromosome instability of the amplified regions. To avoid these problems, we are now building transgene arrays from large genomic DNA inserts cloned in bacterial artificial chromosomes (BAC). These BAC transgenes show transcriptional levels within several fold of endogenous genes while also exhibiting targeting to specific nuclear compartments similar to the targeting of the endogenous genes. Here we describe Tn5 transposition and BAC recombineering methods used to retrofit BACs for their use in building BAC transgene arrays. This includes insertion of operator repeats and selectable markers into these BACs as well as targeted insertion or deletion of BAC sequences.


Quantitation and IgG subclass distribution of antichromatin autoantibodies in SLE mice.
Antibodies to a native chromatin preparation were found in most mice suffering from spontaneous SLE. All MRL/Mp-lpr/lpr (MRL/lpr) sera tested (more than 500) contained antibodies to chromatin and antichromatin levels increased with age. Approximately 50% of the IgG antichromatin antibody in the MRL/lpr sera was of the IgG2a subclass, 30% IgG2b, 10% IgG1, and 10% IgG3. Interestingly, the relative restriction of antichromatin autoantibodies to the IgG2a subclass was apparent in MRL/lpr mice as young as 1 month, well before the onset of lymphadenopathy. Antichromatin autoantibodies were also detectable in sera from MRL/Mp- +/+ (MRL/+), NZB, (NZB x NZW)F1 (B x W), and BXSB mice, but were not found in sera from normal mice. A similar subclass distribution skewed toward IgG2a was seen for MRL/+, B x W, and NZB mice. These results indicate that the spontaneous autoantibody directed against chromatin is a good marker for murine SLE, and is predominantly of the IgG2a subclass.


The efficacy of postoperative autotransfusion in patients undergoing cardiac operations.
The efficacy of postoperative autotransfusion in lowering the requirement for banked-blood transfusion was studied in two groups, each having 168 patients, who underwent cardiac operations between April, 1979, and May, 1980. A Sorenson autotransfusion system was available for use in the autotransfusion group, whereas the control group received routine closed mediastinal drainage. Of the autotransfusion group, 81% met the criterion for autotransfusion (mediastinal losses of 450 ml or more during 4 hours), but only 61% of the autotransfusion group actually received autologous blood (mean autotransfusion volume, 399 +/- 25 ml). The patients receiving autologous blood required significantly less banked blood than their matched controls (447 +/- 60 ml and 744 +/- 83 ml, respectively; p less than 0.001). In the subgroup of patients with large mediastinal losses (more than 1,250 ml), this difference was even greater (autotransfusion, 642 ml compared with control, 1,145 ml; p less than 0.01). Postoperative autotransfusion is a simple, safe, and cost-effective method to reduce dependence on banked blood, especially when mediastinal losses are large. Obtaining maximum benefit requires familiarity of staff with the system and use of a consistent protocol.


Prong sign: a simple measure to enhance safety of laparoscopic sterilization.
Laparoscopic female sterilization is widely practiced and is very safe. However, mesosalpingeal tears and mesosalpingeal haematomas are common complications especially when performed by an inexperienced surgeon. It was found that the observation of a simple 'prong sign' could prevent these complications and possibly reduce failure rates.


Early versus late postural tachycardia: a re-evaluation of a syndrome.
Postural tachycardia syndrome (POTS) involves an HR-rise within 10 minutes of head-up tilt. Hypokinetic circulation, older age, and ACE-inhibitor or Angiotensin-Receptor Blockers were associated with "Late" POTS (after 10 minutes of tilt) versus "Early" POTS (within 10 minutes of tilt).


Roughened titanium surfaces with silane and further RGD peptide modification in vitro.
The strategy to achieve osteoregeneration of dental implants during early-stage regeneration is strongly related to surface conditions for achieving highly successful effects after implantation. Surface modifications, namely, mechanical ground, silanization, bonded and sandblasted with pentasequence Gly-Arg-Gly-Asp-Ser (GRGDS) peptide, and acid-etched with Arg-Gly-Asp (RGD) peptide, were compared for their ability to support cell attachment, proliferation, and differentiation on titanium surfaces. The characteristics and comparative in vitro bio-interactions toward osteoprogenitor cells were tested in the four groups with various surface modifications. Compared with the other groups, the sandblasted and acid-etched, and silane with subsequent RGD peptide modified surfaces had the smallest wetting angle, absence of a significant cell viability difference, and largest quantity of alkaline phosphatase production during the expressions of early-stage cell differentiation. The method of synthesizing GRGDS peptides on roughened titanium surfaces has the potential to provide a combination of early bone regeneration and implant of long-term anchored capabilities.