Visualization of optical binding of microparticles using a femtosecond fiber optical trap.
At the microscopic level, light-matter interactions can organize colloidal matter via a process known as optical binding. Optical binding refers to the creation of arrays of microparticles formed in the presence of laser fields, the inter-particle spacing being determined by the refocusing and/or scattering of the laser fields by the microparticles. In this paper we investigate one-dimensional optically bound arrays of microparticles using a femtosecond dual-beam optical fiber trap, and develop a means to visualize the field intensity distributions responsible for the optical binding using two-photon fluoresence imaging from fluorescein added to the host medium. The experimental intensity distributions are shown to be in good agreement with numerical simulations, thereby validating our new approach to visualizing the fields responsible for optical binding, and the physical model of optical binding as due to refocusing of the fields by the microparticles.


Effects of enterocentesis on peritoneal fluid constituents in the horse.
Peritoneal fluid was collected from 15 clinically normal horses and was analyzed for nucleated cell (NC) counts and specific gravity. Six horses (controls, group 1) were subjected to abdominocentesis only, with a teat cannula, every 24 hours for 5 days. There were no marked changes in the peritoneal fluid of these horses over the 5-day period. Peritoneal fluid was collected from 6 other horses (group 2) with an 8.89-cm 18-gauge needle. The needle was then advanced until intestinal fluid was obtained. Peritoneal fluid was then collected with teat cannulas at 24-hour intervals for an additional 4 days. Peritoneal fluid NC counts from group 2 horses were significantly increased (P less than 0.05) at peak values 2 days after enterocentesis. Specific gravities of peritoneal fluid from group 2 horses were increased on days 1 and 2 after enterocentesis (P greater than 0.05). Peritoneal fluid from 3 other horses (group 3) was collected before enterocentesis (base line) and again at 4-hour intervals after enterocentesis. Peritoneal fluid NC counts of group 3 horses were markedly increased above base-line values 4 hours after enterocentesis and continued to increase for up to 12 hours after enterocentesis when the experiment was terminated. All horses that underwent enterocentesis remained clinically normal except 1 group 3 horse that had a fever (39.6 C) 24 hours after enterocentesis.


Chemokines and chemokine receptors in renal pathology.
Chemokines are members of the largest group of chemotactic cytokines, and were the first shown to be able to engage specific subpopulations of inflammatory cells. Accordingly, our expanding knowledge in chemokine biology has enlarged our understanding of inflammatory cell interactions, lymphopoesis, specificity of cell recruitment, and a variety of human diseases. This review covers recent developments on chemokines in renal diseases. Intrinsic renal cells are capable of chemokine expression in vitro and in vivo, and the involved induction pathways are becoming increasingly defined. Differential chemokine expression during the time course of disease, followed by an infiltration of cells expressing the corresponding receptors has been described in animal models. Therapeutic efficacy of chemokine blockade has been demonstrated in a variety of disease models, including progressive interstitial fibrosis. Chemokine receptors are differentially expressed and localized to specific parenchymal compartments in human renal diseases, as revealed by studies of renal biopsies, and some functional roles of specific chemokine/receptor interactions can be deduced through the correlation of patterns of expression, genetic variations and disease courses. Chemokines play an important role in renal inflammation. Although the treatment of patients with renal diseases using chemokine receptor blocking agents has not yet reached clinical practice, a recent body of data indicates that human renal disease might be amenable to such approaches.


Human cytomegalovirus induces upregulation of arginase II: possible implications for vasculopathies.
Both human cytomegalovirus (HCMV) and arginase II (ARG II) have been implicated in the pathogenesis of cardiovascular diseases. The effects of HCMV on ARG II are unknown. The aim of this study was to investigate the effects of HCMV on ARG II expression in endothelial and vascular smooth muscle cells (SMC) both in vitro and ex vivo. Endothelial and SMC were infected with either HCMV or UV-irradiated HCMV. Expression of ARG II, endothelial or inducible nitric oxide synthase (eNOS and iNOS, respectively) and viral immediate early (IE) was quantified using quantitative PCR. Ganciclovir and short interfering RNA were used to determine the viral gene mediating the effects on ARG II. Detection of viral antigens and ARG II expression was performed by immunofluorescence or immunohistochemistry. HCMV infection increased both ARG II mRNA and protein levels in the examined cells; this effect was mediated by the HCMV IE2-p86 protein. The upregulation of ARG II was accompanied by a downregulation of eNOS but an induction of iNOS in HCMV-infected endothelial cells. Both eNOS and iNOS expressions were induced in HCMV-infected SMC. ARG II was abundantly expressed in endothelial cells, foam cells and SMC and was importantly significantly upregulated in HCMV-immunoreactive human carotid atherosclerotic plaques. HCMV IE2-p86 mediates ARG II upregulation in vitro and ARG II is co-expressed with HCMV antigens in human carotid atherosclerotic plaques. We speculate that HCMV may contribute to endothelial dysfunction via ARG II induction and reduced eNOS production.


The genome sequence of the Large Ear, Amphipoea lucens (Freyer, 1845).
We present a genome assembly from an individual male Amphipoea lucens (the Large Ear; Arthropoda; Insecta; Lepidoptera; Noctuidae). The genome sequence is 647.7 megabases in span. Most of the assembly is scaffolded into 31 chromosomal pseudomolecules, including the assembled Z sex chromosome. The mitochondrial genome has also been assembled and is 15.3 kilobases in length.


Storage of hydrogen spin polarization in long-lived 13C2 singlet order and implications for hyperpolarized magnetic resonance imaging.
Hyperpolarized magnetic resonance imaging (MRI) is a powerful technique enabling real-time monitoring of metabolites at concentration levels not accessible by standard MRI techniques. A considerable challenge this technique faces is the T1 decay of the hyperpolarization upon injection into the system under study. Here we show that A(n)A'(n)XX' spin systems such as (13)C2-1,2-diphenylacetylene ((13)C2-DPA) sustain long-lived polarization for both (13)C and (1)H spins with decay constants of almost 4.5 min at high magnetic fields of up to 16.44 T without spin-locking; the T1 of proton polarization is only 3.8 s. Therefore, storage of the proton polarization in a (13)C2-singlet state causes a 69-fold extension of the spin lifetime. Notably, this extension is demonstrated with proton-only pulse sequences, which can be readily implemented on standard clinical scanners.


[Comparative dynamics of markers of inflammation and NT-proBNP in different variants of treatment of patients with ACS].
STUDY AIM - assessment of dynamics of markers of inflammation (CRP, Il-6, Il-10, TNFa, CD40L, fibrinogen) and N - N in acute coronary syndrome (ACS) as well as analysis of effect of various methods of treatment on level of these parameters. Patient aged 30 - 70 years were included in the study: 52 patients with ACS with ST-segment elevation (STEACS) and 61 - without ST-segment elevation (NSTEACS). Initial level of markers of inflammation (Il-6, CRP) in STEACS was lower than in NSTEACS. Initial level of antiinflammatory Il-10 was significantly higher in patients in the STEACS group (72.6 +/- 39.1 and 6.6+4.2 pg/ml, < 0.01). At admission the highest values of N - N were noted in the group of NSTEACS (761.5 pg/ml compared with 451.1 pg/ml in STEACS, =0.04). During period of hospitalization elevation of N - N occurred in the group of STEACS while its lowering occurred in the group of NSTEACS. In STEACS patients with early restored coronary blood flow dysfunction of the myocardium was less pronounced (lower level of N - N ). In patients with NSTEACS during period of hospitalization levels of CRP, Il-6 fibrinogen lowered. In invasively treated patients with NSTEACS levels of CRP and fibrinogen lowered to a greater extent than in conservatively treated.


Modification of sleep-waking and electroencephalogram induced by vetiver essential oil inhalation.
Essential oils (EOs) have been claimed to modulate mental functions though the most of data were obtained from subjective methods of assessment. Direct effects of EO on brain function remained largely to be confirmed with scientific proof. This study aimed to demonstrate quantifiable and reproducible effects of commercial vetiver (Vetiveria zizanioides) EO inhalation on sleep-waking and electroencephalogram (EEG) patterns in adult male Wistar rats. The experiments were conducted during November 2013 - February 2014. The following electrode implantation on the skull, control, and treated animals were subjected for EEG recording while inhaling water and vetiver EO (20 and 200 µl), respectively. Fast Fourier transform was used for analysis of EEG power spectrum. One-way ANOVA analysis confirmed that vetiver EO inhalation significantly increased total waking and reduced slow-wave sleep time. Moreover, EO inhalation decreased alpha and beta1 activity in both frontal and parietal cortices and increased gamma activity in the frontal cortex. Changes in these frequencies began almost from the start of the inhalation. These data suggest refreshing properties of vetiver EO on electrical brain activity and alertness.


Association of gestational free and total triiodothyronine with gestational hypertension, preeclampsia, preterm birth and birth weight: an individual-participant data meta-analysis.
Triiodothyronine (T3) is the bioactive form of thyroid hormone. In contrast to TSH and FT4, we lack knowledge on the association of gestational T3 with adverse obstetric outcomes. We collected individual participant data from prospective cohort studies on gestational free and/or total T3 (FT3 or TT3), adverse obstetric outcomes (preeclampsia, gestational hypertension, preterm birth and very preterm birth, small for gestational age (SGA) and large for gestational age (LGA)) and potential confounders. We used mixed-effects regression models adjusting for potential confounders. Final study population comprised of 33,118 mother-child pairs of which 27,331 had data on FT3 and 16,164 on TT3. There was a U-shaped association of FT3 with preeclampsia (P = 0.0069) and a J-shaped association with the risk of gestational hypertension (P = 0.029). Higher TT3 was associated with a higher risk of gestational hypertension (OR per SD of TT3 [95% CI]: 1.20 [1.08 to 1.33]; P = 0.0007). A lower TT3 but not FT3 was associated with a higher risk of very preterm birth (OR [95% CI]: 0.72 [0.55 to 0.94]; P = 0.018). TT3 but not FT3 was positively associated with birth weight (Mean difference per 1-SD increase in TT3 [95% CI]: 12.8 [6.5 to 19.1] grams, P < 0.0001) but there was no association with SGA or LGA. This study provides new insights on the association of gestational FT3 and TT3 with major adverse pregnancy outcomes which form the basis for future studies required to elucidate the effects of thyroid function on pregnancy outcomes. Based on the current study, routine FT3 or TT3 measurements for the assessment of thyroid function during pregnancy do not seem to be of added value in the risk assessment for adverse outcomes.


Unexpectedly high levels and health risks of atmospheric polychlorinated biphenyls in modern mechanical dismantling of obsolete electrical equipment: Investigations in a large integrated e-waste dismantling industrial estate.
Large industrial estates for electrical and electronic waste (e-waste) mechanical dismantling and recycling are gradually replacing outmoded small factories and intensive domestic workshops for e-waste manual and chemical dismantling. However, the air pollution and health risks of persistent organic pollutants during the modern mechanical processing of e-waste, especially obsolete electrical equipment, still remain unclear. Here, unexpectedly high levels (409.3 ng/m3) and health risks of airborne polychlorinated biphenyls (PCBs) were found during the mechanical processing of obsolete electric equipment or parts in a large integrated dismantling industrial estate, which is comparable to or a dozen times higher than those reported during chemical processing. In contrast, the levels (936.0 pg/m3) and health risks of particulate polybrominated diphenyl ethers (PBDEs) were all lower than those of previous studies. PCB emissions (44.9-3300.5 ng/m3) varied significantly across six mechanical dismantling places specifically treating waste motors, electrical appliances, hardware, transformers, and metals, respectively. The high PCB content and mass processing number of obsolete electrical equipment probably result in the highest PCB emissions from the mechanical dismantling of obsolete motors, followed by waste electrical appliances and metals. The PCB non-cancer and cancer risks associated with inhalation and dermal exposure in different mechanical dismantling places were all above the given potential risk limits. In particular, the health risks of dismantling obsolete motor exceeded the definite risk levels. Little difference in PCB emissions and health risks between working and non-working time suggested the importance of PCB volatilization from most e-waste. Such high PCB emissions and health risks of PCBs undoubtedly posed a severe threat to frontline workers, but fortunately, they decreased significantly with the increasing distance from the industrial estate. We highlight that PCB emissions and associated health risks from obsolete electrical equipment with high PCB content during mechanical dismantling activities should be of great concern.


Antidepressant and antipsychotic use in an Italian pediatric population.
The safety and effectiveness of psychotropic drug use in the paediatric population is widely debated, in particular because of the lack of data concerning long term effects.In Italy the prevalence of psychotropic drug prescriptions increased in the early 2000s and decreased afterwards. In such a context, a study with the aim to estimate the incidence and prevalence of psychotropic drug prescription in the paediatric population and to describe diagnostic and therapeutic approaches was performed. The study population was composed of 76,000 youths less than 18 years and living in the area covered by the local health unit of Verona, Italy. The data source was the Verona local health unit's administrative prescription database. Prevalence and incidence of antidepressant and/or antipsychotic drug prescriptions in the 2004-2008 period were estimated. Children and adolescents receiving antidepressant and/or antipsychotic drug prescriptions between 1 January 2005 and 31 December 2006 were identified and questionnaires were sent to the prescribers with the aim to collect data concerning diagnostic and therapeutic approaches, and care strategies. The prevalence of psychotropic drug prescriptions did not change in the 2004-2008 period, while incidence slightly increased (from 7.0 in 2005 to 8.3 per 10,000 in 2008). Between 1 January 2005 and 31 December 2006, 111 youths received at least one psychotropic drug prescription, 91 of whom received antidepressants. Only 28 patients attended child and adolescent psychiatry services. Information concerning diagnostic and therapeutic approaches, and care strategies was collected for 52 patients (47%). Anxiety-depressive syndrome and attention disorders were the diseases for which psychotropic drugs were most commonly prescribed. In all, 75% youths also received psychological support and 20% were prescribed drugs for 2 or more years. Despite the low drug prescription prevalence, the finding that most children were not cared for by child and adolescent psychiatric services is of concern and calls for a systematic, continuous monitoring of psychopharmacological treatments.


[Determination of 14 trace elements in soybean and its products by atomic absorption spectrometry].
Fourteen trace elements in soybean and its products were determined by atomic absorption spectrometry. The effects of cinefaction temperature, cinefaction time, and the concentration of HNO3 as a digestion solution were investigated in detail. The effect of the concentration of SrCl2 on the determination of Ca and Mg was also studied. The results obtained show that the soybean and its products contain higher amounts of K, Na, Ca, Mg, Fe, Cu, Zn and Mn than other elements.


Nickel toxicity in wood frog tadpoles: Bioaccumulation and sublethal effects on body condition, food consumption, activity, and chemosensory function.
Nickel (Ni) concentrations in aquatic ecosystems can be amplified by anthropogenic activities including resource extraction. Compared with fish and invertebrates, knowledge of Ni toxicity in amphibians is limited, especially for northern species. We examined the effect of Ni on wood frog (Lithobates sylvaticus) tadpoles, the species with the widest and most northern distribution of any anuran in North America. Wood frog tadpoles were exposed to a Ni concentration gradient (0.02-5.5 mg/L of Ni at 164 mg/L as CaCO3 water hardness) for 8 d and examined for lethality, Ni bioaccumulation, and several sublethal endpoints including body condition, food consumption, activity, and chemosensory function. Nickel induced a sublethal effect on body condition (8-d 10 and 20% effect concentrations [EC10 and EC20] of 1.07 ± 0.38 and 2.44 ± 0.51 mg/L of Ni ± standard error [SE], respectively) but not on food consumption, activity, or chemosensory function. Nickel accumulation in tadpole tissues was positively related to an increase in aqueous Ni concentration but was not lethal. Both the acute and chronic US Environmental Protection Agency water quality guideline concentrations for Ni (0.71 and 0.08 mg/L at 164 mg/L as CaCO3 water hardness, respectively) were protective against lethal and sublethal effects in wood frog tadpoles. In the present study, wood frog tadpoles were protected by current water quality guidelines for Ni and are likely not as useful as other taxa for environmental effects monitoring for this particular metal. Environ Toxicol Chem 2018;37:2458-2466. © 2018 SETAC.


Alu-mediated deletion of PIGL in a Patient with CHIME syndrome.
CHIME syndrome is a rare autosomal recessive neuroectodermal disorder associated with biallelic mutations in PIGL. To date, six molecularly confirmed cases of CHIME syndrome have been reported. Here, we report the seventh patient with biallelic PIGL mutations associated with CHIME syndrome and describe the first characterization of an intragenic deletion in PIGL. Our characterization of the deletion breakpoint junction demonstrated that the breakpoints occurred within Alu repeats and the deletion was most likely mediated by a microhomology event. Analysis of PIGL genomic sequences for repetitive elements demonstrated that Alu repeats represent ∼34% of its intronic sequence, suggesting that the genomic architecture may predispose the gene to disease-causing copynumber changes. Taken together, these findings indicate that patients with a clinical diagnosis of CHIME syndrome and a single identifiable mutation in PIGL warrant further investigation for copynumber changes involving PIGL.


Interference of immunoglobulins in two glucagon radioimmunoassays.
Radioimmunoassays of glucagon in plasma may be complicated by interaction with other substances of high molecular mass. Precipitates of such substances with ammonium sulfate showed, after isoelectric focusing, two fractions having glucagon immunoreactivity. One fraction (pl approximately 10) evidently is associated with the Fc portion (but not the Fab portion) of purified polyclonal immunoglobulin G (IgG). Equal amounts of purified monoclonal IgG of various subclasses, especially IgG 1, gave different "glucagon" readings, suggesting that some IgG may interfere more strongly than others. The other fraction (pI 5-6) appeared less consistently, and on gel chromatography appeared to be slightly larger than IgG. Together these fractions add about 50-100 ng/L to the immunoreactive glucagon values in plasma. Therefore methods in which glucagon is extracted before assay should be used for determining the concentration of glucagon present physiologically.


Afferent visual pathways in multiple sclerosis: a review.
Multiple sclerosis (MS) is a disease of the central nervous system that involves inflammation and demyelination at multiple sites and causes a wide variety of clinical presentations with variable neurological deficits. The visual pathways are frequently involved with either visual or motor dysfunction. Optic neuritis (ON) is one the most common and best characterized presentations of the disease, but there are many other manifestations depending on the site of the lesion. Eyes that have never had ON show slow progressive loss of axons and retinal ganglion cells. Previously unrecognized optic radiation lesions may be associated with residual latency delays on visual evoked potentials. Both anterograde and retrograde degeneration may occur along the visual pathway. This review covers the features of MS in the anterior and posterior visual system and describes advances that have been made with newer techniques such as retinal optical coherence tomography (OCT), magnetic resonance imaging (MRI) with diffusion tensor imaging and probabilistic tractography (DTI) and multifocal visual evoked potentials (mfVEPs). We report on the inter-relationship between these measures of structure and function, and how they may be used as biomarkers for the disease.


Herpesvirus surveillance and discovery in zoo-housed ruminants.
Gammaherpesvirus infections are ubiquitous in captive and free-ranging ruminants and are associated with a variety of clinical diseases ranging from subclinical or mild inflammatory syndromes to fatal diseases such as malignant catarrhal fever. Gammaherpesvirus infections have been fully characterized in only a few ruminant species, and the overall diversity, host range, and biologic effects of most are not known. This study investigated the presence and host distribution of gammaherpesviruses in ruminant species at two facilities, the San Diego Zoo and San Diego Zoo Safari Park. We tested antemortem (blood, nasal or oropharyngeal swabs) or postmortem (internal organs) samples from 715 healthy or diseased ruminants representing 96 species and subspecies, using a consensus-based herpesvirus PCR for a segment of the DNA polymerase (DPOL) gene. Among the 715 animals tested, 161 (22.5%) were PCR and sequencing positive for herpesvirus, while only 11 (6.83%) of the PCR positive animals showed clinical signs of malignant catarrhal fever. Forty-four DPOL genotypes were identified of which only 10 have been reported in GenBank. The data describe viral diversity within species and individuals, identify host ranges of potential new viruses, and address the proclivity and consequences of interspecies transmission during management practices in zoological parks. The discovery of new viruses with wide host ranges and presence of co-infection within individual animals also suggest that the evolutionary processes influencing Gammaherpesvirus diversity are more complex than previously recognized.


A periplasmic LolA derivative with a lethal disulfide bond activates the Cpx stress response system.
LolA accommodates the acyl chains of lipoproteins in its hydrophobic cavity and shuttles between the inner and outer membranes through the hydrophilic periplasm to place lipoproteins in the outer membrane. The LolA(I93C/F140C) derivative, in which Cys replaces Ile at position 93 and Phe at position 140, strongly inhibited growth in the absence of a reducing agent because of the lethal intramolecular disulfide bond between the two Cys residues. Expression of I93C/F140C was found to activate the Cpx two-component system, which responds to cell envelope stress. The inhibition of growth by I93C/F140C was partly suppressed by overproduction of LolCDE, which is an ATP-binding cassette transporter and mediates the transfer of lipoproteins from the inner membrane to LolA. A substantial portion of the oxidized form, but not the reduced one, of I93C/F140C expressed on LolCDE overproduction was recovered in the membrane fraction, whereas wild-type LolA was localized in the periplasm even when LolCDE was overproduced. Moreover, LolCDE overproduction stabilized I93C/F140C and therefore caused an increase in its level. Taken together, these results indicate that oxidized I93C/F140C stably binds to LolCDE, which causes strong envelope stress.


Dose-Response of Sodium Bicarbonate Ingestion Highlights Individuality in Time Course of Blood Analyte Responses.
To defend against hydrogen cation accumulation and muscle fatigue during exercise, sodium bicarbonate (NaHCO3) ingestion is commonplace. The individualized dose-response relationship between NaHCO3 ingestion and blood biochemistry is unclear. The present study investigated the bicarbonate, pH, base excess and sodium responses to NaHCO3 ingestion. Sixteen healthy males (23 ± 2 years; 78.6 ± 15.1 kg) attended three randomized order-balanced, nonblinded sessions, ingesting a single dose of either 0.1, 0.2 or 0.3 g·kg-1BM of NaHCO3 (Intralabs, UK). Fingertip capillary blood was obtained at baseline and every 10 min for 1 hr, then every 15 min for a further 2 hr. There was a significant main effect of both time and condition for all assessed blood analytes (p ≤ .001). Blood analyte responses were significantly lower following 0.1 g·kg-1BM compared with 0.2 g·kg-1BM; bicarbonate concentrations and base excess were highest following ingestion of 0.3 g·kg-1BM (p ≤ .01). Bicarbonate concentrations and pH significantly increased from baseline following all doses; the higher the dose the greater the increase. Large interindividual variability was shown in the magnitude of the increase in bicarbonate concentrations following each dose (+2.0-5; +5.1-8.1; and +6.0-12.3 mmol·L-1 for 0.1, 0.2 and 0.3 g·kg-1BM) and in the range of time to peak concentrations (30-150; 40-165; and 75-180 min for 0.1, 0.2 and 0.3 g·kg-1BM). The variability in bicarbonate responses was not affected by normalization to body mass. These results challenge current practices relating to NaHCO3 supplementation and clearly show the need for athletes to individualize their ingestion protocol and trial varying dosages before competition.


Extrahepatic soft tissue mimicking applicator-tract implantation after percutaneous radiofrequency ablation of hepatic malignancy.
The purpose of our study was to identify extrahepatic soft tissue mimicking applicator-tract implantation after percutaneous radiofrequency ablation of hepatic malignancy on triple-phase dynamic CT. A database was reviewed for 131 patients who underwent percutaneous radiofrequency ablation for treatment of a hepatic tumor over a 3-year period. Patients who developed extrahepatic soft tissue adjacent to the previous ablation zone mimicking applicator-tract implantation were identified. The clinical features of patients, characteristics of extrahepatic soft tissue on CT, soft-tissue changes on follow-up CT, and histopathologic results were evaluated. Extrahepatic soft tissue developed in five patients (3.8%) at 2-8 months after percutaneous radiofrequency ablation. Extrahepatic soft tissue showed progressive enhancement during the delayed phase of dynamic CT. Four lesions were completely resolved without any treatment. However, one lesion was enlarged and xanthogranulomatous inflammation was diagnosed by percutaneous biopsy. Univariate analysis showed that an index tumor with a subcapsular location and a prior biopsy had high odds ratios. The possibility of a benign inflammatory lesion should be considered when extrahepatic soft tissue showing radiologic findings similar to applicator-tract implantation develops after radiofrequency ablation of a hepatic tumor.


Absolute configuration of aflastatin A, a specific inhibitor of aflatoxin production by Aspergillus parasiticus.
Aflastatin A (1) is a specific inhibitor of aflatoxin production by Aspergillus parasiticus. It has the novel structure of a tetramic acid derivative with a long alkyl side chain. The absolute configurations of 29 chiral centers contained in 1 were chemically elucidated in this study. First, four small fragment molecules were prepared from 1 or its methyl ether (2), and their absolute structures were assigned as N-methyl-D-alanine, (2S,4R)-2, 4-dimethyl-1,6-hexanediol dibenzoate, (R)-3-hydroxydodecanoic acid, and (R)-1,2,4-butanetriol tribenzoate. Next, an acyclic fragment molecule 3 with 13 chiral centers was obtained from 1 by NaIO(4) oxidation, and its relative stereochemistry was elucidated by J-based configuration analysis. By analyzing coupling constants of (3)J(H,H) and (2,3)J(C,H) and ROE data, the relative configuration of 3 was verified. Finally, by further J-based configuration analysis using a fragment molecule 7 prepared from 2 with 28 chiral carbons, all relative configurations in the alkyl side chain of 1 were clarified. By connecting these relative configurations with the absolute configurations of first four fragment molecules, the absolute stereochemistry of 1 was fully determined.


Hydration Differences Explain the Large Variations in the Complexation Thermodynamics of Modified γ-Cyclodextrins with Bile Salts.
The structure and thermodynamics of inclusion complexes of seven different γ-cyclodextrins (γCDs) and three biologically relevant bile salts (BS) were investigated in the present study. Natural γCD and six modified γCDs [two methyl-γCDs, one sulfobutyl ether-γCD (SBEγCD), and three 2-hydroxypropyl-γCDs (HPγCD)] and their complexes with BS were investigated by isothermal titration calorimetry, NMR, and molecular dynamics simulations. With the exception of the fully methylated γCD, which did not bind the BSs investigated, all of the γCDs formed 1:1 complexes with the BS, and the structures were similar to those with natural γCD; i.e., the modifications of the γCD had limited structural impact on the formation of complexes. Isothermal titration calorimetry was carried out over in the temperature interval 5-55 °C to enable the calculation of the stability constant (K) and the thermodynamic parameters enthalpy (ΔH°), entropy (ΔS°), and heat capacity (ΔCp°). The stability constants decreased with an increased degree of substitution (DS), with methyl substituents having a lower effect on the stability constant than the sulfobutyl ether and hydroxypropyl substituents on the stability constants. Enthalpy-entropy compensation was observed, since both enthalpy and entropy increased with the degree of substitution, which may reflect dehydration of the hydrophobic surface on both CD and BS. Calculations based on ΔCp° data suggested that each of the substituents dehydrated 10-20 (hydroxypropyl), 22-33 (sulfobutyl ether), and 10-15 Å(2) (methyl) of the BS surface area, in reasonable agreement with estimates from the molecular dynamics simulations. Combined with earlier investigations on modified βCDs, these results indicate general trends of the substituents on the thermodynamics of complex formation.


Effect of two dietary concentrate levels on tenderness, calpain and calpastatin activities, and carcass merit in Waguli and Brahman steers.
The objective of this study was to compare carcass characteristics of a newly introduced breed, the Waguli (Wagyu x Tuli), with the carcass characteristics of the Brahman breed. Brahman cattle are used extensively in the Southwest of the United States because of their tolerance to adverse environmental conditions. However, Brahman carcasses are discounted according to the height of their humps because of meat tenderness issues. The Waguli was developed in an attempt to obtain a breed that retained the heat tolerance of the Brahman but had meat quality attributes similar to the Wagyu. Twenty-four animals were used. Six steers from each breed were fed a 94% concentrate diet and 6 steers from each breed were fed an 86% concentrate diet. Eight steers, 2 from each group, were harvested after 128 d, after 142 d, and after 156 d on feed. Waguli steers had larger LM, greater backfat thickness, greater marbling scores, and greater quality grades than the Brahman steers (P < 0.05). The Japanese Wagyu breed is well known for its highly marbled and tender meat, and these traits are also present in the Waguli. The Waguli had significantly lower Warner-Bratzler shear force values than the Brahman steers after 7 and 10 d of postmortem aging (P < 0.05); this difference decreased after 14 d postmortem (P = 0.2), when tenderness of the slower aging Brahman had increased to acceptable levels. Toughness of the Brahman has been associated with high levels of calpastatin in Brahman muscle, and the Waguli LM had significantly less calpastatin activity (P = 0.02) at 0 h postmortem than the Brahman LM. At 0-h postmortem, the total LM calpain activity did not differ between the Brahman and Waguli (P = 0.57). Neither diet nor days on feed had any significant effect on the 0-h postmortem calpain or at 0-h postmortem calpastatin activity, nor an effect on Warner-Bratzler shear-force values. In conclusion, LM muscle from the Waguli steers had a high degree of marbling, lower shear force values, and low calpastatin activity, all of which are related to more tender meat.


Predictive value of non-specific bronchial challenge testing for respiratory symptoms and lung function in aluminium smelter workers.
To assess the predictive value of bronchial hyper-responsiveness (BHR) for the subsequent development of respiratory symptoms, airflow limitation and decline in lung function among aluminium smelter workers. An inception cohort study of new employees at two Australian aluminium smelters was conducted. Participants completed a modified British Medical Research Council respiratory questionnaire, spirometry and a methacholine bronchial challenge test at baseline and at annual follow-up reviews. BHR was defined as PD20 ≤4000 µg. Poisson and mixed effects models were fitted to respiratory symptoms and lung function (forced expiratory volume in 1 s (FEV1) and FEV1/forced vital capacity (FVC)). Baseline interview and lung function testing were completed by 278 workers, who were followed for a median of 4 years. BHR at baseline, present in 82 workers, was not associated with incident wheeze risk ratio (RR)=1.07 (95% CI 0.74 to 1.55) and cough RR=0.78 (95% CI 0.45, 1.35), but there was some increased risk of chest tightness RR=1.40 (95% CI 0.99, 1.98) after adjustment for age, sex, smoking and atopy. BHR at baseline was associated with lower FEV1 and FVC, although the rate of annual decline in FEV1 or FVC was similar between those with or without BHR. The specificity of BHR was 77% for wheeze, 70% for cough and 77% for chest tightness, but the sensitivity was poor, at 33%, 24% and 39%, respectively. Methacholine challenge testing at entry to employment was not sufficiently predictive of later adverse respiratory outcomes, and notwithstanding the study limitations is unlikely to be a useful pre-employment or preplacement screening test in the aluminium smelting industry.


Albumin-to-Alkaline Phosphatase Ratio Associates with Good Prognosis of Hepatitis B Virus-Positive HCC Patients.
The aim of this study was to investigate the prognostic significance of preoperative AAPR in hepatitis B virus-related hepatocellular carcinoma patients after curative hepatectomy. A total of 221 patients with hepatitis B virus-related HCC patients who received curative liver resection were included. After propensity matching analysis, 188 patients were enrolled in the final analysis. COX regression analyses were used to analyze the prognosis value of AAPR and other prognostic factors. The overall survival (OS) and recurrence-free survival (RFS) curves were constructed and compared between different groups. The optimal cutoff of AAPR was defined as 0.40 with X-tile software. According to cutoff value, patients were divided into low-AAPR group (≤0.40) and high-AAPR group (>0.40). The cumulative 1-, 3-, and 5-year OS rates were 97.1%, 78.2%, and 67.3% in patients with AAPR>0.40 group, respectively, which were significantly higher than those in the AAPR≤0.40 group (80.2%, 54.4%, and 40.1%, respectively) (P <0.001). In the multivariate COX regression analysis, AAPR, tumor number, ascites, and portal vein tumor thrombus (PVTT) were independent risk factors for OS and RFS. AAPR shows promise as a reliable prognostic factor in patients with hepatitis B virus-related HCC after curative hepatectomy, which could be used as a routine inspection of HCC patients before surgery.


Cubosomes: The Next Generation of Smart Lipid Nanoparticles?
Cubosomes are highly stable nanoparticles formed from the lipid cubic phase and stabilized by a polymer based outer corona. Bicontinuous lipid cubic phases consist of a single lipid bilayer that forms a continuous periodic membrane lattice structure with pores formed by two interwoven water channels. Cubosome composition can be tuned to engineer pore sizes or include bioactive lipids, the polymer outer corona can be used for targeting and they are highly stable under physiological conditions. Compared to liposomes, the structure provides a significantly higher membrane surface area for loading of membrane proteins and small drug molecules. Owing to recent advances, they can be engineered in vitro in both bulk and nanoparticle formats with applications including drug delivery, membrane bioreactors, artificial cells, and biosensors. This review outlines recent advances in cubosome technology enabling their application and provides guidelines for the rational design of new systems for biomedical applications.


Phospholipase A2 inhibitors and leukotriene synthesis inhibitors block TNF-induced NF-kappaB activation.
Tumour necrosis factor (TNF) is a key regulator of inflammation and immunity. The cellular effects exerted by TNF depend, apart from NF-kappaB-directed gene transcription, largely on its ability to activate phospholipase A2(PLA2), yielding the release of arachidonic acid (AA) and its metabolites. AA metabolites, especially the leukotrienes, act as second messengers in TNF receptor signalling, as different inhibitors of AA metabolism impair a variety of TNF-induced biochemical events. The role, however, of AA and its metabolites in TNF-induced NF-kappaB activation is still obscure. Here we report that 4-bromophenacyl bromide (4-BPB; an inhibitor of PLA2), nordihydroguaretic acid (NDGA; a 5-lipoxygenase inhibitor), as well as MK-886 [an inhibitor of 5-lipoxygenase-activating protein (FLAP)] interfere with TNF-induced NF-kappaB-mediated transactivation. However, only 4-BPB inhibited the DNA-binding activity of NF-kappaB, whereas NDGA and MK-886 did not. Thus, different inhibitors interfere at different points in TNF-induced signalling leading to NF-kappaB-dependent transcription. Artificial induction of AA metabolism induced neither DNA-binding activity of NF-kappaB nor NF-kappaB-dependent transactivation. It was concluded that although TNF-induced signalling to NF-kappaB-dependent transcription is sensitive to inhibitors of AA metabolism at multiple points during this signalling, AA release is essential but not sufficient for NF-kappaB-activation.


[Clinical experience in using a metallic suture in eye microsurgery].
The use of a metallic suture in eye microsurgery is described in the paper. The metallic suture made from a 0.05-0.05 mm vanadic stainless steel wire (corresponding to 8:0) was applied in 1202 cataract extractions, penetrating injuries, antiglaucomatous surgeries and in removal of pterygium at the medical-and-sanitary facility of steel works in the town of Bokaro (India). The observation period ranged, on the average, from 3 months to 3 years. There were no complications during the procedure of applying the suture. However, it caused a minor response in the ocular tissues during the early postoperative period. Small bullae were observed in the corneal surface near the suture in 7 cases (0.58%), which disappeared spontaneously in 3-4 days. Irritations inflicted by the suture ends were registered in 2.58% of cases (31 patients). The mean postoperative astigmatism was, 3 months after surgery, 1.65 diopters. The sutures were removed in 132 cases (10.98%), including in 16 patients with irritation, and in 116 patients for the purpose of correcting a high astigmatism. The share of patients with a visual acuity of 0.5 and higher was, in one week, 43.59% and went up, one month later, to 78.20%. The suture was well tolerated by patients, it was quickly covered by the corneal epithelium, there was no rejection and it can be easily manufactured. The suture is sterilized by routine techniques without need for any extra treatment before use; it is a reactive, biologically inactive, efficient, convenient and causes minimal operative and postoperative complications.


Fast normalized cross correlation for motion tracking using basis functions.
Digital image-based elasto-tomography (DIET) is an emerging method for non-invasive breast cancer screening. Effective clinical application of the DIET system requires highly accurate motion tracking of the surface of an actuated breast with minimal computation. Normalized cross correlation (NCC) is the most robust correlation measure for determining similarity between points in two or more images providing an accurate foundation for motion tracking. However, even using fast Fourier transform (FFT) methods, it is too computationally intense for rapidly managing several large images. A significantly faster method of calculating the NCC is presented that uses rectangular approximations in place of randomly placed landmark points or the natural marks on the breast. These approximations serve as an optimal set of basis functions that are automatically detected, dramatically reducing computational requirements. To prove the concept, the method is shown to be 37-150 times faster than the FFT-based NCC with the same accuracy for simulated data, a visco-elastic breast phantom experiment and human skin. Clinically, this approach enables thousands of randomly placed points to be rapidly and accurately tracked providing high resolution for the DIET system.


Lost but not forgotten: A population-based study of mortality and care trajectories among people living with HIV who are lost to follow-up in Ontario, Canada.
Selection as a consequence of volunteer participation in, and loss to follow-up from, cohort studies may bias estimates of mortality and other health outcomes. To quantify this potential, we estimated mortality and health service use among people living with HIV (PLWH) who were lost to cohort follow-up (LTCFU) from a volunteer clinical HIV-infected cohort, and compared these to mortality and health service use in active cohort participants and non-cohort-participants living with HIV in Ontario, Canada. We analysed population-based provincial health databases from 1995 to 2014, identifying PLWH ≥ 18 years old; these included data from participants in the Ontario HIV Treatment Network Cohort Study (OCS), a volunteer, multi-site clinical HIV-infected cohort. We calculated all-cause mortality, hospitalization and emergency department (ED) visit rates per 100 person-years (PY) and estimated hazard ratios (HRs) of mortality, adjusting for age, sex, income, rurality, and immigration status. Among 23 043 PLWH, 5568 were OCS participants. Compared with nonparticipants, participants were younger and less likely to be female, to be an immigrant and to reside in a major urban centre, and had lower comorbidity. Mortality among active participants, participants LTCFU and nonparticipants was 2.52, 3.30 and 2.20 per 100 PY, respectively. After adjustment for covariates, mortality risk was elevated among participants LTCFU compared with active participants (HR 2.26; 95% confidence interval 1.91, 2.68). Age-adjusted hospitalization rates and ED visit rates were highest among participants LTCFU. Mortality risk and use of health care resources were lower among active cohort participants. Our findings may inform health outcome estimates based on volunteer cohorts, as well as quantitative bias adjustment to correct for such biases.


Megabase-scale mapping of the HLA gene complex by pulsed field gel electrophoresis.
In the study of the genetic structure of mammalian chromosomes, there exists a "resolution gap" between molecular cloning experiments and meiotic linkage analyses. This gap has discouraged attempts to construct full-scale genetic maps of mammalian chromosomes. The organization of the human major histocompatibility complex was examined within this range by pulsed field gel electrophoresis. The data obtained indicate that the complex spans over 3000 kilobases and enable the construction of a megabase-scale molecular map. These results indicate that the techniques employed in DNA extraction, enzymatic digestion, electrophoresis, and hybridization are suitable for the efficient analysis of megabase regions of mammalian chromosomes and effectively bridge the resolution gap between molecular cloning and classical genetics.


Studies on the ability to taste phenylthiocarbamide; red-green colour blindness and the age at menarche in Johannesburg Chinese school children.
The sample, although small, represents the total number of children attending the Johannesburg Chinese Kuo Ting Primary and High School in 1972 to 1974. The frequency of the non-taster gene was found to be 0,167 in the combined male and female sample. The gene for red-green colour blindness is present in the Johannesburg Chinese population but at low frequnecy, 0,012. The mean age at menarche was found to be 12,8+/-1,23 years and is younger than the mean ages recorded for a number of Caucasoid populations. It is suggested that this difference in mean age reflects possibly varying genetic rather than varying socio-economic factors.


Effects of biopsychosocial factors on the association between loneliness and mental health risks during the COVID-19 lockdown.
Previous data suggest that loneliness is a hallmark of the mental health issues triggered by the SARS-CoV-2 pandemic. The purpose of the present study was to examine the impact of loneliness on mental health and behavioural outcomes, as well as the moderating and mediating effects of biopsychosocial variables on these relationships. The data were collected during France's first national COVID-19 lockdown and included 556 participants (M age = 30.06 years, range = 18-87) who completed well-known validated measures assessing symptom-levels of anxiety, depression, and insomnia. They also indicated their level of worry about the COVID-19 crisis, and provided sociodemographic and health status data. Responses were submitted to hierarchical linear regression and mediation analyses. In terms of prevalence of loneliness, 18.9% of participants reported severe loneliness. High levels of loneliness were significantly associated with the three mental health and behavioural outcomes. The loneliness - anxiety relationship was moderated by employment and working arrangements. The loneliness - insomnia relationship was moderated by living conditions, history of medical or psychological problems, and COVID-19-related worry. Further findings supported the indirect effects of COVID-related worry on pathways from loneliness to the three mental health and behavioural health outcomes. The mediation models accounted for 28.9%, 33.7%, and 15.0% of the variance in anxiety, depression, and insomnia. The present evidence-based findings revealed that loneliness and worry were major contributing factors of mental health and behavioural concerns during the COVID-19 lockdown. They could inform treatment recommendations for tackling prolonged-self isolation and loneliness in specific vulnerable groups. The online version contains supplementary material available at 10.1007/s12144-021-02246-w.


[Properties of synaptic currents during non-adrenergic inhibition of the smooth muscle cells of the large intestine in the guinea pig].
Inhibitory junctional currents (IJCs) were recorded under voltage clamp conditions in response to brief transmural stimulation of the circular muscle of the guinea pig colon using the double sucrose gap method in the presence of atropine. The time course of IJC decay was approximately exponential 100-150 ms after the peak value. The IJC amplitude depended linearly on the membrane potential with the reversal potential (-70 mV) near the potassium equilibrium potential. The time constant (tau) of the IJC decay depended exponentially on the membrane potential and became e-fold decreased when the membrane was hyperpolarized approximately by 120 mV. Varying the quantal content of IJC caused an increase of tau upon rising the amount or transmitter released and its decrease with the depression of IJC. Application of ATP (10(-3)M) caused a decrease of tau and IJC amplitude, while apamine reduced the amplitude of IJC without any changes in their time course. The results are discussed in terms of a buffered diffusion hypothesis supposing a cooperative action of transmitter released on junctional receptors.


Online support for caregivers. Analysis of an Internet Alzheimer mailgroup.
Alzheimer's disease affects more than four million Americans and ranks as the fourth leading cause of death in adults. To deal with the stresses of constant caregiving, family members of patients with Alzheimer's disease are encouraged to participate in support groups. However, geographic and time constraints combined with lack of substitute caregivers may preclude attendance at traditional support groups. Internet-based groups offer caregivers the opportunity to interact with other caregivers for guidance, information, and encouragement. This project examines the content and themes of 532 messages posted on a public Alzheimer mailgroup during 20 days of 1998. Members used the mailgroup to find and proffer information, share experiences and opinions, and provide encouragement. By becoming aware of the potential advantages that online support groups offer to caregivers, nurses can encourage Alzheimer caregivers to use online groups as an empowering and practical adjunct to traditional caregiver support.


Intracellular tension in periosteum/perichondrium cells regulates long bone growth.
Perichondrium/periosteum is involved in regulating long bone growth. Long bones grow faster after removal or circumferential division of periosteum. This can be countered by culturing them in conditioned medium from perichondrium/periosteum cells. Because both complete removal and circumferential division are effective, we hypothesized that perichondrium/periosteum cells require an intact environment to release the appropriate soluble factors. More specifically, we propose that this release depends on their ability to generate intracellular tension. This hypothesis was explored by modulating the ability of perichondrium/periosteum cells to generate intracellular tension and monitoring the effect thereof on long bone growth. Perichondrium/periosteum cells were cultured on substrates with different stiffness. The medium produced by these cultures was added to embryonic chick tibiotarsi from which perichondrium/periosteum was either stripped or left intact. After 3 culture days, long bone growth was proportionally related to the stiffness of the substrate on which perichondrium/periosteum cells were grown while they produced conditioned medium. A second set of experiments demonstrated that the effect occurred through expression of a growth-inhibiting factor, rather than through the reduction of a stimulatory factor. Finally, evidence for the importance of intracellular tension was obtained by showing that the inhibitory effect was abolished when perichondrium/periosteum cells were treated with cytochalasin D, which disrupts the actin microfilaments. Thus, we concluded that modulation of long bone growth occurs through release of soluble inhibitors by perichondrium/periosteum cells, and that the ability of cells to develop intracellular tension through their actin microfilaments is at the base of this mechano-regulated control pathway.


Regeneration of irradiation-damaged esophagus by local delivery of mesenchymal stem-cell spheroids encapsulated in a hyaluronic-acid-based hydrogel.
Radiation therapy (RT) is a typical treatment for head and neck cancers. Generally, prolonged irradiation of the esophagus causes esophageal fibrosis due to increased reactive oxygen species and proinflammatory cytokines. This study was designed to determine whether catechol-functionalized hyaluronic acid (HA-CA) hydrogel-encapsulated human mesenchymal stem-cell spheroids (MSC-SPs) could ameliorate damage to the esophagus in a mouse model of radiation-induced esophageal fibrosis. MSC-SPs were cultured in concave microwells 600 μm in diameter at a cell density of 1 × 106 cells per mL. Most cells formed spheroids with a 100-300 μm size distribution in concave microwells. MSC-SPs were well maintained in the HA gel, and live-dead staining confirmed that most cells survived. The HA gel containing the MSC-SPs was then injected into the damaged esophageal layer. Inflammatory signs or adverse tissue reactions were not observed after esophageal injection of HA-gel-encapsulated MSC-SPs. Based on Masson's trichrome staining at 4 and 12 weeks postinjection, the inner esophageal layer (IEL) was significantly thinner in the MSC-SP + HA gel group compared to those in the other experimental groups. While the saline and HA gel treatments made the esophageal muscles loose and thick, the MSC-SP + HA gel group showed bundles of tightly packed esophageal muscles, as assayed by desmin immunostaining. qPCR analysis showed that epithelial genes tended to increase over time in the MSC-SP + HA gel group, and the expression of most fibrosis-related genes decreased. This study proposes the potential of using HA-CA-hydrogel-encapsulated MSC-SPs as a promising therapy against radiation-induced esophageal fibrosis.


Mathematical structures of line drawings of polyhedrons-toward man-machine communication by means of line drawings.
Mathematical structures of line drawings of polyhedrons are studied and practical as well as theoretical solutions are obtained for several fundamental problems aroused in scene analysis and in man-machine communication. First, a necessary and sufficient condition for a line drawing to correctly represent a polyhedron is obtained in terms of linear algebra. Next, combinatorial structures are investigated and practical solutions are obtained to such problems as how to discriminate between correct and incorrect line drawings and how to correct vertex-position errors in incorrect line drawings. Lastly, distribution of the degree of freedom of a line drawing is elucidated and a method is proposed for interactive reconstruction of a polyhedron from a line drawing. The results obtained here enable us to make manmachine communication more ``flexible'' in the sense that a machine can reconstruct three-dimensional objects from hand-drawn pictures even if the pictures are not perfect.


The feasibility of a porcine model of acute coronary occlusion and reperfusion using off-pump coronary artery bypass grafting.
We developed an open-chest porcine model of acute coronary occlusion and surgical reperfusion, and attempted to prevent intra-operative ischaemic ventricular fibrillation (VF) by a Retrograde Intracoronary Glyceryl trinitrate (RIG) infusion into the occluded vessel. Five Yorkshire pigs (weight 50 +/- 1.1 kg), randomized into 3 groups, underwent median sternotomy under general anaesthesia. One pig (Group 1, control) underwent sternotomy and pericardiotomy only. Four pigs underwent acute left anterior descending (LAD) coronary occlusion. Two pigs were not reperfused (Group 2). Two pigs underwent surgical reperfusion (Group 3) via left internal mammary artery (LIMA) grafting to the LAD using the Off-Pump Coronary Artery Bypass (OPCAB) technique. Ischaemic injury was assessed using 7-lead electrocardiography (ECG) and transthoracic/epimyocardial echocardiography (ECHO). Group 1: transient intraoperative hypotension and VF occurred. Successful resuscitation and 10-week survival (until sacrifice) with normal left ventricular (LV) function was achieved. Group 2: there were ECG and ECHO evidence of acute LV ischaemic dysfunction in both pigs. The surviving pig had persistent anterior hypokinesis at 8 1/2 months. The other died intra-operatively following progressive ischaemic LV dysfunction despite resuscitative attempts. Group 3: the surviving pig had normal LV function at 8 months. Initial anterior LV akinesis normalized within 7 days. The other developed post-occlusion haemodynamic instability and died intra-operatively despite reperfusion. In this porcine model, acute LAD artery occlusion modified by the novel RIG infusion technique, followed by surgical reperfusion (OPCAB) is feasible. This model would facilitate further development of OPCAB surgical expertise and understanding of the pathophysiology of ischaemia-reperfusion injury.


Management of adjuvant settings for Stage III melanoma patients in France prior to checkpoint inhibitors: epidemiological data from the RIC-Mel database.
Targeted therapies such as BRAF and MEK inhibitors and immunotherapies have been made available to treat melanoma. To provide an overview of the management of the French Stage III melanoma population after complete lymph node resection prior to new adjuvant therapies. A subgroup data analysis. Data from 1,835 patients were analysed (15.58% Stage IIIA, 39.24% Stage IIIB, 43.92% Stage IIIC and 1.25% Stage IIID). Superficial spreading melanoma was the most frequent (70.98% in Stage IIIA for whom mutation analysis was performed; BRAF mutation was identified in up to 62% Stage IIIA patients). Sentinel lymph node biopsy was performed in 88.46% of Stage IIIA patients, 42.36% of Stage IIIB, 53.97% of Stage IIIC and 34.78% of Stage IIID. Up to 80% of Stage IIIA patients had no adjuvant treatment follow-up. Ulceration (p = 0.004; RR: 2.98; 95% CI: 1.4-6.3) and age at diagnosis (p = 0.0002; RR: 1.04; 95% CI: 1.02-1.06) were significant predictive factors for survival. Adjuvant interferon-α was administered in up to 13.04% of Stage IIID patients. Only a small number of Stage III melanoma patients were treated with interferon-α in adjuvant settings. New adjuvant therapies are currently having an effect on clinical practice in France, increasing survival and decreasing cost.


hnRNP H is a component of a splicing enhancer complex that activates a c-src alternative exon in neuronal cells.
The regulation of the c-src N1 exon is mediated by an intronic splicing enhancer downstream of the N1 5' splice site. Previous experiments showed that a set of proteins assembles onto the most conserved core of this enhancer sequence specifically in neuronal WERI-1 cell extracts. The most prominent components of this enhancer complex are the proteins hnRNP F, KSRP, and an unidentified protein of 58 kDa (p58). This p58 protein was purified from the WERI-1 cell nuclear extract by ammonium sulfate precipitation, Mono Q chromatography, and immunoprecipitation with anti-Sm antibody Y12. Peptide sequence analysis of purified p58 protein identified it as hnRNP H. Immunoprecipitation of hnRNP H cross-linked to the N1 enhancer RNA, as well as gel mobility shift analysis of the enhancer complex in the presence of hnRNP H-specific antibodies, confirmed that hnRNP H is a protein component of the splicing enhancer complex. Immunoprecipitation of splicing intermediates from in vitro splicing reactions with anti-hnRNP H antibody indicated that hnRNP H remains bound to the src pre-mRNA after the assembly of spliceosome. Partial immunodepletion of hnRNP H from the nuclear extract partially inactivated the splicing of the N1 exon in vitro. This inhibition of splicing can be restored by the addition of recombinant hnRNP H, indicating that hnRNP H is an important factor for N1 splicing. Finally, in vitro binding assays demonstrate that hnRNP H can interact with the related protein hnRNP F, suggesting that hnRNPs H and F may exist as a heterodimer in a single enhancer complex. These two proteins presumably cooperate with each other and with other enhancer complex proteins to direct splicing to the N1 exon upstream.


The Use of Primary Care Electronic Health Records for Research: Lipid Medications and Mortality in Elderly Patients.
General practice electronic health record (EHR) data have significant potential for clinical research. This study demonstrates the feasibility of utilising longitudinal EHR data analysis to address clinically relevant outcomes and uses the relationship between lipid medication prescription and all-cause mortality in the elderly as an exemplar for the validity of this methodology. EHR data were analysed to describe the association of lipid medication use, non-use or cessation with all-cause mortality in patients aged ≥75 years. Survival analysis with Cox regression was used to calculate hazard ratios, which were adjusted for confounders. There was no significant difference in all-cause mortality among patients according to their use, non-use, or cessation of lipid medications. The outcomes of this study correlate well with the results of other research works. This single-practice study demonstrates the feasibility and potential of analysing EHR data to address important clinical issues such as the relationship between all-cause mortality and lipid medication prescription in the elderly.


Sleep-disordered breathing in acute ischemic stroke and transient ischemic attack: effects on short- and long-term outcome and efficacy of treatment with continuous positive airways pressure--rationale and design of the SAS CARE study.
Sleep-disordered breathing represents a risk factor for cardiovascular morbidity and mortality and negatively affects short-term and long-term outcome after an ischemic stroke or transient ischemic attack. The effect of continuous positive airways pressure in patients with sleep-disordered breathing and acute cerebrovascular event is poorly known. The SAS CARE 1 study assesses the effects of sleep-disordered breathing on clinical evolution, vascular functions, and markers within the first three-months after an acute cerebrovascular event. The SAS CARE 2 assesses the effect of continuous positive airways pressure on clinical evolution, cardiovascular events, and mortality as well as vascular functions and markers at 12 and 24 months after acute cerebrovascular event. SAS CARE 1 is an open, observational multicenter study in patients with acute cerebrovascular event acutely admitted in a stroke unit: a sample of 200 acute cerebrovascular event patients will be included. Vascular functions and markers (blood pressure, heart rate variability, endothelial function by peripheral arterial tonometry and specific humoral factors) will be assessed in the acute phase and at three-months follow-up. SAS CARE 2 will include a sample of patients with acute cerebrovascular event in the previous 60-90 days. After baseline assessments, the patients will be classified according to their apnea hypopnea index in four arms: non-sleep-disordered breathing patients (apnea hypopnea index <10), patients with central sleep-disordered breathing, sleepy patients with obstructive apnea hypopnea index ≥20, which will receive continuous positive airways pressure treatment, nonsleepy patients with obstructive sleep-disordered breathing (apnea hypopnea index ≥20), which will be randomized to receive continuous positive airways pressure treatment or not. The SAS CARE study will improve our understanding of the clinical sleep-disordered breathing in patients with acute cerebrovascular event and the feasibility/efficacy of continuous positive airways pressure treatment in selected patients with acute cerebrovascular event and sleep-disordered breathing.


Anal skin tags in inflammatory bowel disease: new observations and a clinical review.
The association between intestinal Crohn's disease (CD) and specific perianal abnormalities called anal skin tags (AST) has been recognized but not well defined. Skin tags have been classified into 2 types: 1) raised, broad, or narrow, single or multiple, soft or firm, and painless, often referred to as "elephant ears"; or 2) edematous, hard, often cyanotic, tender or not, and typically arising from a healed anal fissure, ulcer, or hemorrhoid. The aims of this study were to i) better characterize those skin tags identified by the term "elephant ears" and differentiate them from other types of AST; ii) compare their prevalence in patients with CD and ulcerative colitis (UC); iii) observe the relationship of the skin tags to the location of the primary bowel disease; and iv) to discuss the value of these typical AST in making an early diagnosis of CD. Photographs of all AST were taken when present at lower endoscopy in 170 consecutive patients with inflammatory bowel disease (IBD) seen in the private office of the senior investigator and Lenox Hill Hospital. Data was gathered with respect to major differences between the 2 types of AST. The location of the primary bowel disease for these patients was obtained from an extensive IBD computer database and review of details from charts. Specific features of AST were described and served to favor type 1 versus type 2. AST were found more frequently in patients with CD (75.4%) as compared to patients with UC (24.6%), confirming previous observations that they are more diagnostic of CD (P = 0.005). Subset analysis revealed a trend with a greater incidence of AST in patients with colitis (46.9%) as compared to patients with ileitis (36.7%) and ileocolitis (16.3%) (P = 0.067). We provide photographs with the most characteristic features of AST and attempt to separate elephant ears (type 1) from less typical AST (type 2) in CD. Our study confirms previous reports that AST are more commonly found in association with CD as compared with UC and more so in the presence of disease limited to the colon as compared to disease elsewhere in the bowel. Our observations support the diagnostic significance of AST heralding the diagnosis of CD when they are discovered on physical exam, especially in young people with diarrhea, abdominal pain, and/or growth retardation.


Quantitative model of the image of a radiating dipole through a microscope.
In this paper, we introduce a formalism to determine the relationship between the full vectorial electric field existing at the object plane of a microscope and that existing at the image plane. The model is then used to quantitatively simulate, in both phase and intensity, the image of a radiating electric dipole placed either in a homogeneous medium or in the vicinity of a substrate. These simulations are compared with experimental measurements on single gold nanoparticles carried out by quadriwave lateral shearing interferometry.


Actions of neurohypophysial peptides on pancreatic hormone release.
The intermittent reports concerning metabolic actions of neurohypophysial extracts or hormones encouraged us to study the effect of these substances on the function of the endocrine pancreas. A surprisingly small amount of neural lobe (NL) extract (0.025 NL eq/ml) stimulated a 425% increase in the release of glucagon from islets isolated from the pancreas of the rat. Gel filtration of the extract produced an elution profile of glucagon-releasing activity that was superimposable on the profiles of oxytocin (OT) and arginine vasopressin (AVP). Synthetic OT and AVP each elicited a concentration-dependent stimulation of glucagon release but failed to influence insulin release in medium 199 containing 5.6 mM glucose. They were effective at 0.2 ng/ml (+55%, +50%) and produced a striking increase (five- to sevenfold) at 20 ng/ml. The response to each peptide was greatly diminished in the presence of a higher concentration of glucose (11 mM). The lysine, desamino-, and 1,6-aminosuberyl analogues of vasopressin, vasotocin, and AVP are equipotent peptides, whereas the desglycinamide analogue, pressinoic acid, and angiotensin II were inactive. Injection of AVP (1 microgram iv) produced a rapid increase in peripheral glucagon (+185% in 5 min). The response to injection of OT was less rapid (+105% in 15 min), but in each case elevation of insulin was also observed. Our results provide evidence that OT and AVP can act directly on the endocrine pancreas and may help explain previous reports of metabolic actions of these peptides.


Epidemiology and virology of acute respiratory infections during the first year of life: a birth cohort study in Vietnam.
Understanding viral etiology and age-specific incidence of acute respiratory infections in infants can help identify risk groups and inform vaccine delivery, but community-based data is lacking from tropical settings. One thousand four hundred and seventy-eight infants in urban Ho Chi Minh City and 981 infants in a semi-rural district in southern Vietnam were enrolled at birth and followed to 1 year of age. Acute respiratory infection (ARI) episodes were identified through clinic-based illness surveillance, hospital admissions and self-reports. Nasopharyngeal swabs were collected from infants with respiratory symptoms and tested for 14 respiratory pathogens using multiplex reverse transcription-polymerase chain reaction. Estimated incidence of ARI was 542 and 2691 per 1000 infant-years, and hospitalization rates for ARI were 81 and 138 per 1000 infant-years, in urban and semi-rural cohorts, respectively, from clinic- and hospital-based surveillance. However self-reported ARI episodes were just 1.5-fold higher in the semi-rural versus urban cohort, indicating that part of the urban-rural difference was explained by under-ascertainment in the urban cohort. Incidence was higher in infants ≥6 months of age than <6 months, but this was pathogen-specific. One or more viruses were detected in 53% (urban) and 64% (semi-rural) of samples from outpatients with ARI and in 78% and 66% of samples from hospitalized ARI patients, respectively. The most frequently detected viruses were rhinovirus, respiratory syncytial virus, influenza virus A and bocavirus. ARI-associated hospitalizations were associated with longer stays and more frequent ICU admission than other infections. ARI is a significant cause of morbidity in Vietnamese infants and influenza virus A is an under-appreciated cause of vaccine-preventable disease and hospitalizations in this tropical setting. Public health strategies to reduce infant ARI incidence and hospitalization rates are needed.


[Genetic counseling in congenital long QT syndrome].
Long QT syndrome is an inherited heart-rhythm disorder characterized by an increased risk of ventricular tachycardia and sudden death. Genetic testing is available. The article is based on an anonymous family with a history of long QT syndrome, the authors experience with this patient group and a Pubmed search for literature from the period 1957 - 2007. An 8-year-old boy suffers syncope at a sports event, and this leads to genetic counseling and molecular genetic testing of his first-and second-degree relatives. Knowledge about genetic risk of sudden death in a family can trigger genetic testing and health preventive treatment of children, but can also have substantial psychosocial and ethical consequences for the family and for the health-care personnel involved. Living with a genetic risk can be very emotionally challenging for the individuals and families, and "The Norwegian Act of Biotechnology in Human Medicine, etc" that regulates clinical genetic activities is extensive. An important question is whether the current Act allows communication of genetic information to persons other than the patient.


Underlying selection for the diversity of spike protein sequences of SARS-CoV-2.
The global spread of SARS-CoV-2 is fast moving and has caused a worldwide public health crisis. In the present article, we analyzed spike protein sequences of SARS-CoV-2 genomes to assess the impact of mutational diversity. We observed from amino acid usage patterns that spike proteins are associated with a diversity of mutational changes and most important underlying cause of variation of amino acid usage is the changes in hydrophobicity of spike proteins. The changing patterns of hydrophobicity of spike proteins over time and its influence on the receptor binding affinity provides crucial information on the SARS-CoV-2 interaction with human receptor. Our results also show that spike proteins have evolved to prefer more hydrophobic residues over time. The present study provides a comprehensive analysis of molecular sequence data to consider that mutational variants might play a crucial role in modulating the virulence and spread of the virus and has immediate implications for therapeutic strategies.


Attenuated baroreflex control of sympathetic nerve activity after cardiovascular deconditioning in rats.
The effect of cardiovascular deconditioning on baroreflex control of the sympathetic nervous system was evaluated after 14 days of hindlimb unloading (HU) or the control condition. Rats were chronically instrumented with catheters and sympathetic nerve recording electrodes for measurement of mean arterial pressure (MAP) and heart rate (HR) and recording of lumbar (LSNA) or renal (RSNA) sympathetic nerve activity. Experiments were conducted 24 h after surgery, with the animals in a normal posture. Baroreflex function was assessed using a logistic function that related HR and LSNA or RSNA to MAP during infusion of phenylephrine and nitroprusside. Baroreflex influence on HR was not affected by HU. Maximum baroreflex-elicited LSNA was significantly reduced in HU rats (204 +/- 11.9 vs. 342 +/- 30.6% baseline LSNA), as was maximum reflex gain (-4.0 +/- 0.6 vs. -7.8 +/- 1.3 %LSNA/mmHg). Maximum baroreflex-elicited RSNA (259 +/- 10.8 vs. 453 +/- 28.0% baseline RSNA), minimum baroreflex-elicited RSNA (-2 +/- 2.8 vs. 13 +/- 4.5% baseline RSNA), and maximum gain (-5.8 +/- 0.5 vs. -13.6 +/- 3.1 %RSNA/mmHg) were significantly decreased in HU rats. Results demonstrate that baroreflex modulation of sympathetic nervous system activity is attenuated after cardiovascular deconditioning in rodents. Data suggest that alterations in the arterial baroreflex may contribute to orthostatic intolerance after a period of bedrest or spaceflight in humans.


Comparison of Inflammatory Cytokine Levels in Hepatic and Jugular Veins of Patients with Cirrhosis.
Systemic inflammation with elevated inflammatory cytokines is a hallmark in patients with cirrhosis and the main driver of decompensation. There is insufficient data on whether inflammatory cytokine levels differ between hepatic and jugular veins, which may have implications for further immunological studies. Blood from the hepatic and jugular veins of 40 patients with cirrhosis was collected during hepatic venous pressure gradient (HVPG) measurements. Serum levels of 13 inflammatory cytokines (IL-1β, Int-α2, Int-γ, TNF-α, MCP-1, IL-6, IL-8, IL-10, IL-12p70, IL-17A, IL-18, IL-23, and IL-33) were quantified by cytometric bead array. Cytokine levels of IFN-α2, IFN-γ, TNF-α, IL-6, IL-8, IL-10, IL-17A, IL-18, IL-23, and IL-33 were significantly elevated in patients with decompensated cirrhosis compared to patients with compensated cirrhosis. When comparing patients with clinically significant portal hypertension (CSPH, HVPG ≥ 10 mmHg) to patients without CSPH, there were significantly enhanced serum levels of IL-6 and IL-18 in the former group. There was no significant difference between cytokine serum levels between blood obtained from the jugular versus hepatic veins. Even in subgroup analyses stratified for an early cirrhosis stage (Child-Pugh (CP) A) or more decompensated stages (CP B/C), cytokine levels were similar. Cytokine levels increase with decompensation and increasing portal hypertension in patients with cirrhosis. There is no relevant difference in cytokine levels between hepatic and jugular blood in patients with cirrhosis.


Postictal agitation after electroconvulsive therapy: incidence, severity, and propofol as a treatment option.
Postictal agitation (PIA) after electroconvulsive therapy (ECT) is a major medical problem. This observational study investigated the incidence and severity of PIA and evaluated propofol as a treatment option in a patient population. The study included 14 patients that underwent a series of ECTs performed either with or without an approximately 0.5-mg/kg propofol bolus after the end of an electroencephalography (EEG) seizure. Among other values, we documented PIA incidence and severity as rated by a simple score; orientation to person, time, place, and situation; transfer times to the postanesthesia care (PACU) and inpatient unit; nurses' and patients' rating of recovery period, and others and tested for significant differences. Five minutes after the end of ECT, the patients showed moderate to severe PIA in 8 of 37 ECT sessions. Incidence was significantly lower when patients had received propofol (3/37). Transfer time to the PACU was longer, but transfer time to the inpatient unit was shorter after administration of propofol. The recovery period was rated significantly better after propofol administration by nurses and patients. A single bolus of propofol administered after the end of the seizure reduced the incidence of post-ECT PIA. The PACU staff and patients rated the emergence period significantly better when propofol was administered.


Case of delayed diagnosis of necrolytic migratory erythema.
Necrolytic migratory erythema (NME) is a rare cutaneous paraneoplastic manifestation of glucagonoma. We report a case of a woman with a 6-year history of delayed diagnosis of glucagonoma. This case highlights the atypical clinical features of NME which makes the diagnosis difficult.


Radical surgery for cervical carcinoma: experience with "the Tokyo method".
Thirty patients of cervical carcinoma were treated by radical abdominal hysterectomy and pelvic and lower paraaortic lymphadenectomy between November 1986 and October 1988 by the 'Tokyo method.' Blood loss in the majority of the patients (80%) was less than 1000 ml. The incidence of intra and postoperative complications was found to be low. None of the patients developed urinary fistula and the incidence of bladder dysfunction was also low (3.3%). Follow up of the patients for nine to 32 months revealed local recurrent tumour in only one case. We feel that the "Tokyo method" of radical hysterectomy offers several advantages over other methods.


Motion velocity thresholds in deaf signers: changes in lateralization but not in overall sensitivity.
In a series of three experiments, we tested whether deaf native signers process motion velocity information differently from hearing nonsigners. In Experiment 1, participants watched radially moving dots and were asked to detect the quadrant in which the velocity of the dots had changed. Similar 79% thresholds were observed in the two populations. In Experiments 2 and 3, peripheral and central thresholds were assessed separately as previous studies suggest early deafness leads mainly to changes in the processing of visual peripheral information. Neither condition produced an overall population difference. These negative results were not due to a lack of sensitivity in our experiments. Indeed, as has been previously reported, deaf native signers exhibited better thresholds in the right than in the left visual field, whereas the opposite pattern was observed in the hearing. This effect appears triggered by experience with American Sign Language (ASL) rather than deafness per se. Overall, this study confirms that early deafness does not enhance motion processing, and suggests that most of the changes previously described in the literature are instead attributable to changes in attention, and possibly special alterations of attention-to-motion processes.


Effect of Flavonoid-rich Extract of Tamarix Articulata Vahl. on Glucose and Lipid Metabolism in Normal and Diabetic Rats.
The present study was carried out in order to evaluate the antidiabetic and antihyperlipidemic potential of Flavonoid-Rich Extract of Tamarix articulata (FRETA) in both normal and Streptozotocin (STZ)-induced diabetic rats. Normal and diabetic rats were treated with the FRETA for 7 days. At the end of treatment, a range of parameters was tested including blood lipid profile, histopathological changes in both liver and pancreas and Oral Glucose Tolerance Test (OGTT). The blood glucose levels were lowered in both normal and diabetic rats treated with FRETA. Single oral administration of FRETA reduced blood glucose levels significantly in both normal and diabetic rats six hours after administration (P < 0.001; P < 0.0001 respectively). Furthermore, blood glucose levels were decreased significantly (P < 0.0001) in diabetic rats after 7 days of treatment. According to the oral glucose tolerance test, FRETA (5 mg/kg) was shown to prevent significantly the increase in blood glucose levels in diabetic treated rats. In addition, FRETA (5 mg/kg) showed a strong hypolipidemic effect both in normal and STZ rats after 7 days of once daily oral treatment. FRETA induced a significant decrease of plasma triglyceride and total cholesterol concentrations both in normal and diabetic rats. In contrast, plasma HDL-c levels were increased significantly (P < 0.0001) both in normal and diabetic rats. In addition, FRETA showed a remarkable in vitro antioxidant activity and revealed the inhibitory concentration of 50% of free radicals (IC50) of 31.92 µg/ml. In diabetic rats, flavonoids from Tamarix articulata showed antidiabetic and hypolipidemic activities.


[Liver disease in toxic oil syndrome. Clinical and enzymatic course].
One-hundred and fifty-nine patients intoxicated with adulterated oil in the province of Salamanca, Spain, were followed-up for 2 years from the onset of the syndrome, in 1981. Four alcoholic patients were excluded from the study. In the remaining 155 patients the liver involvement at the onset took different forms. It was acute in 12 (7%), with jaundice and/or hepatomegaly associated with cytolysis and/or cholestasis; 21 patients (13%) had evidence of cytolysis and/or cholestasis without clinical symptoms, and 29 (18%) only showed a rise in gamma-glutamyltranspeptidase with normal transaminase and alkaline phosphatase levels. Liver enlargement and/or jaundice subsided within 1 month in the 12 patients with clinical symptoms. However, high gamma-glutamyltranspeptidase levels persisted in 4 of them and in 5 of the 21 patients with laboratory abnormalities only. At the end of the study, this enzyme remained elevated in 6 and 7 patients respectively of these two groups and in 5 of the 29 patients with no other abnormality than a rise in gamma-glutamyltranspeptidase.


Factors determining outcomes in grown up patients operated for congenital heart diseases.
The number of grown ups with congenital heart diseases (GUCHs) is steadily increasing. To analyze factors predicting early cardiac morbidity following cardiac surgery in GUCH at a tertiary care center. Retrospective study at a multispeciality tertiary referral center. Between January 2004 and December 2014, 1432 patients ≥13 years of age (acyanotic defects: 843, cyanotic defects: 589) underwent surgery for congenital heart defects. Factors associated with early cardiac morbidity were analyzed. Univariable and multivariable analysis of all factors affecting outcomes. On multivariate analysis, previous sternotomy, aortic cross-clamp time >45 min, cyanosis, and emergency procedure were independent predictors of early morbidity with respective odds ratios (ORs) of 12.4, 3.6, 2.6, and 8.1. For more precise estimation, a risk score was generated. Taking the log odds with each of these four as respective weights, a score was generated. The variables were previous sternotomy (2.5), aortic cross-clamp >45 min (1.3), emergency procedure (2.1), and cyanosis (0.9), if the respective condition is present, zero otherwise. The score ranged from 0 to 4.5. The average value of the score based on the four variables was significantly higher in cases with morbidity (1.85 ± 1.17) vs. (0.75 ± 0.88), P < 0.001. Distribution of scores was significantly different between patients with and without morbidity. Sixty-seven percent patients without any morbidity had score <1 compared to 24.6% with morbidity. Only 0.9% patients without morbidity had score of ≥3 compared to 16.4% patients with morbidity. Compared with patients having score <1, patients with scores 1-2 had OR of 3.4, 2-3 had OR of 6.0, and >3 had OR of 48.7. GUCH can be safely operated when adequate caution is taken in the presence of independent predictors such as previous sternotomy, aortic clamp time >45 min, cyanosis, and emergency procedure.


Early and late effect of infliximab on circulating dendritic cells phenotype in rheumatoid arthritis patients.
The aim of this study was to analyze the phenotype of circulating dendritic cells (DCc) in rheumatoid arthritis (RA) patients before and after treatment with infliximab (at 24 h and 6 months) and the correlation between these changes and the clinical response to treatment. Sixteen patients with RA were recruited and clinical status was determined using the Disease Activity Score 28 (DAS28). All patients had active disease (mean DAS28 = 5.96) and were suitable for treatment with infliximab. Samples of peripheral venous blood were obtained before administration of the first dose of infliximab and again at 24 h and 6 months after treatment. DCc populations were analyzed by flow cytometry. At 24 h, there were no differences in the clinical status of the patients. However, we found a decrease in CD11c+ and, to a lesser extent, CD123+ DCc percentages. The expression of CD83, the most important activation marker for DC, was also shown to be decreased 24 h after infliximab therapy. After 6 months of treatment, all patients showed significant clinical improvement (mean DAS28 = 3.64, p < 0.001) and expression of the activation marker on DCc remained low. In conclusion, this study supports the role of tumor necrosis factor (TNF)-alpha blockade in preventing the maturation of DCc and in reducing the expression of their activation markers. Although the clinical response to infliximab was not observed after 24 h, DCc activation was strongly reduced by anti-TNF-alpha therapy. After 6 months of treatment, current data show a less active phenotype of DCc associated with clinical improvement in all patients in the study.


Development and evolution of the vertebrate primary mouth.
The vertebrate oral region represents a key interface between outer and inner environments, and its structural and functional design is among the limiting factors for survival of its owners. Both formation of the respective oral opening (primary mouth) and establishment of the food-processing apparatus (secondary mouth) require interplay between several embryonic tissues and complex embryonic rearrangements. Although many aspects of the secondary mouth formation, including development of the jaws, teeth or taste buds, are known in considerable detail, general knowledge about primary mouth formation is regrettably low. In this paper, primary mouth formation is reviewed from a comparative point of view in order to reveal its underestimated morphogenetic diversity among, and also within, particular vertebrate clades. In general, three main developmental modes were identified. The most common is characterized by primary mouth formation via a deeply invaginated ectodermal stomodeum and subsequent rupture of the bilaminar oral membrane. However, in salamander, lungfish and also in some frog species, the mouth develops alternatively via stomodeal collar formation contributed both by the ecto- and endoderm. In ray-finned fishes, on the other hand, the mouth forms via an ectoderm wedge and later horizontal detachment of the initially compressed oral epithelia with probably a mixed germ-layer derivation. A very intriguing situation can be seen in agnathan fishes: whereas lampreys develop their primary mouth in a manner similar to the most common gnathostome pattern, hagfishes seem to undergo a unique oropharyngeal morphogenesis when compared with other vertebrates. In discussing the early formative embryonic correlates of primary mouth formation likely to be responsible for evolutionary-developmental modifications of this area, we stress an essential role of four factors: first, positioning and amount of yolk tissue; closely related to, second, endoderm formation during gastrulation, which initiates the process and constrains possible evolutionary changes within this area; third, incipient structure of the stomodeal primordium at the anterior neural plate border, where the ectoderm component of the prospective primary mouth is formed; and fourth, the prime role of Pitx genes for establishment and later morphogenesis of oral region both in vertebrates and non-vertebrate chordates.


Birth weight of offspring and subsequent cardiovascular mortality of the parents.
Restricted fetal growth has been associated with increased blood pressure, insulin resistance, and cardiovascular disease in later life. These associations may be due to processes during fetal development with long-term consequences for metabolic and cardiovascular function (the fetal origins hypothesis). However, it has also been hypothesized that common genetic factors could underlie both restricted fetal development and disease risk (the fetal insulin hypothesis). We have investigated associations between offspring size at birth and parental mortality in a record linkage study including all 783,814 children born in Sweden between 1973 and 1980. The deaths of 10,368 mothers and 22,807 fathers had been recorded by date and causes of death. Data were analyzed using Cox proportional hazards regression models, with parents' age as the time axis. Offspring birth weight (adjusted for gestational age) was inversely associated with all-cause mortality and cardiovascular mortality in the parents, with weaker associations for fathers than for mothers. The inverse association between birth weight and cardiovascular disease mortality was greatest for parents who died at younger ages. Cancer and alcohol-related causes of death were also related to offspring birth weight. A sensitivity analysis suggests that confounding by smoking is unlikely to entirely explain the associations. Associations between offspring birth characteristics and parents' mortality could, at least in part, reflect genetic factors that influence both birth weight and cardiovascular disease risk.


[Basic procedure for blood cultures (author's transl)].
The Author summarizes the basic routine procedure for blood cultures first examining different factors that can influence it (blood coagulation, antimicrobial agents, natural bacterial inhibitors of blood cell wall deficient bacteria) and then describing culture media, atmospheric conditions, incubation time, and modalities for inspection, staining of cultures and subcultures. Finally new rapid automated methods are briefly pointed out.


Safety of treatment.
Data concerning safety of treatment in schoolchildren cannot be extrapolated to preschool children due to differences in growth velocity and metabolism. The safety issue in preschool children is further complicated by insufficient knowledge of the optimal dose, and the lung delivery from the devices available. Systemic activity has often been studied as a marker of adverse clinical effect. However, with improving technology, systemic activity can be detected within the range of the normal biological feedback system, where it is of no clinical importance. Therefore, systemic activity is not synonymous with clinically relevant side-effects. Side-effects should be assessed in specific clinical studies. Effect on growth velocity is a potential side-effect of major interest. Knemometry is a sensitive measure of short-term growth of the lower leg length in schoolchildren as well as in preschool children, which enables precise measurements of systemic activity, but it is not a measure of statural growth. The only clinically relevant outcome measure of human growth is the final height in relation to expected final height, allowing for gender and midparental height differences. In addition to effects on statural height, osteoperosis is an important potential side-effect. The importance of bone density during preschool age for final adult bone mineral density needs to be studied. Cataract formation is a side-effect associated with systemic steroid treatment and may be of special consideration during treatment with nebulized steroids, which may expose the eyes to high doses. Thrush, dysphonia and local skin atrophy in steroid-exposed areas are potential local side-effects, and yet the incidence in young children is unknown and dependent on the device used for delivery. When considering adverse effects of treatment, the risk of side-effects from undertreatment should always be observed.


EFFECT OF TEMPERATURE ON THE COMPOSITION OF FATTY ACIDS IN ESCHERICHIA COLI.
Marr, Allen G. (University of California, Davis) and John L. Ingraham. Effect of temperature on composition of fatty acids in Escherichia coli. J. Bacteriol. 84:1260-1267. 1962.-Variations in the temperature of growth and in the composition of the medium alter the proportions of individual fatty acids in the lipids of Escherichia coli. As the temperature of growth is lowered, the proportion of unsaturated fatty acids (hexadecenoic and octadecenoic acids) increases. The increase in content of unsaturated acids with a decrease in temperature of growth occurs in both minimal and complex media. Cells harvested in the stationary phase contained large amounts of cyclopropane fatty acids (methylenehexadecanoic and methylene octadecanoic acids) in comparison with cells harvested during exponential growth. Cells grown in a chemostat, limited by the concentration of ammonium salts, show a much higher content of saturated fatty acids (principally palmitic acid) than do cells harvested from an exponentially-growing batch culture in the same medium. Cells grown in a chemostat, limited by the concentration of glucose, show a slightly higher content of unsaturated fatty acids than cells from the corresponding batch culture. The results do not indicate a direct relation between fatty acid composition and minimal growth temperature.


The association between systemic inflammation and cognitive performance in the elderly: the Sydney Memory and Ageing Study.
Inflammation may contribute to cognitive decline and dementia. This study examined the cross-sectional relationships between markers of systemic inflammation (C-reactive protein, interleukins-1β, -6, -8, -10, -12, plasminogen activator inhibitor, serum amyloid A, tumour necrosis factor-α and vascular adhesion molecule-1) and cognitive function in 873 non-demented community-dwelling elderly participants aged 70-90 years. Regression analyses were performed to determine the relationships between cognitive domains and inflammatory markers, controlling for age, sex, education, cardiovascular risk factors, obesity and other metabolic factors, smoking, alcohol consumption, depression and presence of the apolipoprotein ε4 genotype. Regression analyses were repeated using four factors derived from a factor analysis of the cognitive tests. After Bonferroni correction for multiple testing, associations remained between raised levels of interleukin-12 and reduced performance in processing speed. Marked sex differences were noted in the abovementioned findings, with only females being significantly affected. Using the four factors derived from the factor analyses of cognitive test as dependent variables, interleukins-12 and -6 were both associated with the processing speed/executive function factor, even after controlling for relevant confounding factors. Thus, markers of systemic inflammation are related to cognitive deficits in a non-clinical community-dwelling elderly population, independent of depression, cardiovascular or metabolic risk factors, or presence of apolipoprotein ε4 genotype. Additional research is required to elucidate the pathophysiology and longitudinal development of these relationships.


Enabling tools for high-throughput detection of metabolites: Metabolic engineering and directed evolution applications.
Within the Design-Build-Test Cycle for strain engineering, rapid product detection and selection strategies remain challenging and limit overall throughput. Here we summarize a wide variety of modalities that transduce chemical concentrations into easily measured absorbance, luminescence, and fluorescence signals. Specifically, we cover protein-based biosensors (including transcription factors), nucleic acid-based biosensors, coupled enzyme reactions, bioorthogonal chemistry, and fluorescent and chromogenic dyes and substrates as modalities for detection. We focus on the use of these methods for strain engineering and enzyme discovery and conclude with remarks on the current and future state of biosensor development for application in the metabolic engineering field.