Serum C-reactive Protein and Protein Electrophoretic Pattern Correlated With Age in Horses.
Inflammaging or chronic, low-grade inflammation is a phenomenon characterizing age-related pathologies and natural processes in aging tissues. This study aimed to investigate the effect of age on the serum levels of C-reactive protein and protein electrophoretic pattern in horses. Thirty Italian Saddle horses from the same horse training center, were enrolled and divided in three equal groups according to their age: Group A (2-4 years), Group B (7-10 years), Group C (15-20 years). Blood samples were collected from each horse to evaluate hematological parameters and the serum concentration of C-reactive protein (CRP), total proteins, albumin, α1-, α-2, β1-, β2- and γ-globulins. One-way analysis of variance showed an age-related difference in the concentration of CRP, α1- and α2-globulins (P < 0.001) which increased with aging. Albumin and albumin:globulin ratio showed lower values in Groups B and C than Group A (P < 0.001). Age of horses showed a negative correlation with the values of albumin, and a positive correlation with CRP, α1- and α2-globulins concentration. The results of the current study suggest an increased acute phase response in adult and old horses compared to young ones. As α-globulin fractions include many other acute phase proteins (APPs) in addition to CRP, further studies to assess what other APPs could be involved in chronic inflammation or "inflammaging" are recommended in horse in order to improve knowledge on the key inflammatory biomarkers during aging in this species.


Potential mucolytic agents for mucinous ascites from pseudomyxoma peritonei.
Pseudomyxoma peritonei is a disease characterised by the accumulation of mucinous ascites. Thus far, cytoreductive surgery combined with hyperthermic intraperitoneal chemotherapy (HIPEC) has been shown to be effective at eradicating disease. Chemotherapy has been less effective, providing disease stabilization but not demonstrating significant treatment responses. Mucolytic is a potential class of drug that may be exploited in the chemical management of this disease. A variety of potential mucolytic agents are explored in this review providing evidence of basic biochemical evidence of its efficacy with potential translational application.


Impact of Eimeria meleagrimitis and intermittent amprolium treatment on performance and the gut microbiome composition of Turkey poults.
Drug-sensitive live coccidiosis vaccines have been used to control coccidiosis and renew drug sensitivity in commercial chicken operations. However, only limited species coverage vaccines have been available for commercial turkey producers. This study aimed to assess the effect of an E. meleagrimitis vaccine candidate, with and without amprolium intervention, on performance and oocyst shedding. Additionally, the effect of vaccination, amprolium treatment, and E. meleagrimitis challenge on intestinal integrity and microbiome composition was evaluated. Experimental groups included: (1) NC (non-vaccinated, non-challenged control); (2) PC (non-vaccinated, challenged control); (3) VX + Amprol (E. meleagrimitis candidate vaccine + amprolium); and 4) VX (E. meleagrimitis candidate vaccine). For VX groups, 50% of the direct poults were orally vaccinated at DOH with 50 sporulated E. meleagrimitis oocysts and were comingled with contact or non-vaccinated poults for the duration of the study. From d10-14, VX + Amprol group received amprolium (0.024%) in the drinking water. All groups except NC were orally challenged with 95K E. meleagrimitis sporulated oocysts/mL/poult at d23. At d29, ileal and cecal contents were collected for 16S rRNA gene-based microbiome analysis. VX did not affect performance during the pre-challenge period. At d23-29 (post-challenge), VX groups had significantly (P < 0.05) higher BWG than the PC group. Contacts and directs of VX groups in LS had significantly reduced compared to PC. As anticipated, amprolium treatment markedly reduced fecal and litter OPG for the VX + Amprol group compared to the VX group which did not receive amprolium. The ileal and cecal content results showed that the PC group had different bacterial diversity and structure, including alpha and beta diversity, compared to NC. Linear discriminant analysis Effect Size (LEfSe) identified that Lactobacillus salivarius (ASV2) was enriched in PC's ileal and cecal content. Compared to NC and PC, the vaccinated groups showed no distinct clusters, but there were similarities in the ileal and cecal communities based on Bray-Curtis and Jaccard distances. In conclusion, these results indicate that vaccination with this strain of E. meleagrimitis, with or without amprolium intervention, caused a very mild infection that induced protective immunity and challenge markedly affected both the ileal and cecal microbiome.


SLC2A9 is a newly identified urate transporter influencing serum urate concentration, urate excretion and gout.
Uric acid is the end product of purine metabolism in humans and great apes, which have lost hepatic uricase activity, leading to uniquely high serum uric acid concentrations (200-500 microM) compared with other mammals (3-120 microM). About 70% of daily urate disposal occurs via the kidneys, and in 5-25% of the human population, impaired renal excretion leads to hyperuricemia. About 10% of people with hyperuricemia develop gout, an inflammatory arthritis that results from deposition of monosodium urate crystals in the joint. We have identified genetic variants within a transporter gene, SLC2A9, that explain 1.7-5.3% of the variance in serum uric acid concentrations, following a genome-wide association scan in a Croatian population sample. SLC2A9 variants were also associated with low fractional excretion of uric acid and/or gout in UK, Croatian and German population samples. SLC2A9 is a known fructose transporter, and we now show that it has strong uric acid transport activity in Xenopus laevis oocytes.


BiClamp® forcep liver transection versus clamp crushing technique for liver resection: study protocol for a randomized controlled trial.
Blood loss and the requirement of blood transfusions during liver transection have been shown to correlate well with higher morbidity and mortality rates and a worse prognosis. Various devices for liver parenchymal transection have been developed to reduce intraoperative blood loss. The goal of this study is to evaluate the safety and effectiveness of BiClamp® forcep transection compared to a clamp crushing technique in patients undergoing liver resection. This study will include patients 18 years and older scheduled for hepatectomy with hepatic vascular exclusion who give informed consent. A sample size of 48 patients in each randomization arm will be calculated to detect a difference in the reduction of blood loss of approximately 200 ml (90% power and α = 0.05 (two-tailed)). The primary efficacy endpoint of the trial will be the total intraoperative blood loss based on the randomized dissection technique. The statistical analysis is based on the intention-to-treat population. Patients will be followed up on for three months for complications and adverse events. This prospective, single-center, randomized controlled, single-blinded, two-group parallel trial is designed to assess the efficacy and safety of BiClamp forcep hepatectomy versus clamp crushing for parenchymal transection during elective hepatic resection. This trial was registered with Clinicaltrials.gov (identifier: NCT02197481 ) on 15 July 2014.


MOSAICING OF DYNAMIC MESENTERY VIDEO WITH GRADIENT BLENDING.
In biomedical imaging using video microscopy, understanding large tissue structures at cellular and finer resolution poses many image acquisition challenges including limited field-of-view and tissue dynamics during imaging. Automated mosaicing or stitching of live tissue video microscopy enables the visualization and analysis of subtle morphological structures and large scale vessel network architecture in tissues like the mesentery. But mosacing can be challenging if there are deformable, motion-blurred, textureless, feature-poor frames. Feature-based methods perform poorly in such cases for the lack of distinctive keypoints. Standard single block correlation matching strategies might not provide robust registration due to deformable content. In addition, the panorama suffers if there is motion blur present in a sequence. To handle these challenges, we propose a novel algorithm, Deformable Normalized Cross Correlation (DNCC) image matching with RANSAC to establish robust registration. Besides, to produce seamless panorama from motion-blurred frames we present gradient blending method based on image edge information. The DNCC algorithm is applied on Frog Mesentery sequences. Our result is compared with PSS/AutoStitch [1, 2] to establish the efficiency and robustness of the proposed DNCC method.


Physician modeling influences on patient smokers.
Physicians may be considered important models of healthful behavior for their patients. This analogue study examined the behavioral impact of advice to quit given by a smoking and nonsmoking physician model on 27 light (less than 20 cigarettes per day) and 27 heavy (greater than 20 cigarettes per day) inpatient smokers in a V.A. Medical Center setting. Results indicated that heavy smokers smoked significantly more than light smokers on all measures across conditions. Interestingly, both heavy and light smokers tended to smoke more when the model advised quitting regardless of the presence or absence of model smoking. Post hoc analyses of locus of control revealed that patients with an internal locus of control tended to react to the physician's advice to quit by smoking significantly more during exposure, irrespective of model behavior. Patients with an external locus of control seemed to respond in a manner consistent with the behavior of the model.


Effects of the mineral composition and water content of intact plants on the fitness of the African armyworm.
The effects of organic nitrogen, nitrate, phosphorus, potassium and water content of leaves of intact maize plants, grown in a gravel culture system, on the fitness of the African armyworm, Spodoptera exempta (Walker)(Lepidoptera: Noctuidae) were studied. Organic nitrogen concentrations ranged from 1.3% to 3.7% over four treatments differing only in nitrate supply to the plants. Water content and other mineral levels were all positively correlated with the organic nitrogen level. Feeding damage by the caterpillars was most severe on the lowest nitrate treatments, where it could be least well compensated for by new leaf growth. Larval and pupal fitness variables were not affected by treatment, except for larval development on the lowest nitrate treatment which was delayed by just 1 day. The large compensatory capacity of the larvae was underlined by a similar mineral composition of the pupae in all treatments. Adult fitness variables hardly differed between the upper three nitrate treatments, but revealed a trend over all treatments: the higher the organic nitrogen content of the leaves, the shorter the pre-oviposition period and the higher the fecundity. This trend, however, might have been due to differences in available food quantity rather than in food quality. It is concluded that fitness of the African armyworm is only slightly affected over a wide range of nitrogen concentrations in its food. Though effects might be larger under field conditions, the large differences in outbreak development between years seem not to be attributable to observed differences in nitrogen levels in host plants between years in primary outbreak areas. Other environmental factors appear to be of greater importance.


Arrhythmias in Type 2 Diabetes Mellitus.
Chronic hyperglycaemia of Type 2 diabetes mellitus causes long term damage to heart resulting in coronary artery disease (CAD), myocardial infarction (MI), congestive heart failure (CHF), and sudden death from arrhythmias. To study the prevalence of different types of arrhythmias in T2DM, particularly in association with Cardiac Autonomic Neuropathy (CAN). A cross-sectional study including 100 patients of Type 2 Diabetes Mellitus (T2DM) presenting with cardiac arrhythmias, was done at our hospital over 2 years. Detailed history along with physical examination and tests for CAN were done. Routine investigations along with echocardiography, stress test, Holter monitoring were done. Sinus Tachycardia (ST) was the commonest arrhythmia, found in 32% of patients. 20% had Complete Heart Block (CHB), 15% had Sinus Bradycardia (SB), and 15% had Atrial Fibrillation (AF). Ventricular Premature Complex (VPC) was found in 10% and 3% had Atrial Premature Complex (APC). 3% had first degree AV block, whereas 1% had Paroxysmal Supra Ventricular Tachycardia (PSVT), and another 1% had Ventricular Tachycardia (VT). Poorly controlled diabetes and co-morbidities was associated with higher incidence of arrhythmias. 62% of patients had prolonged QTc, majority of which had CAN. Most of the patients responded to standard therapy.


Knockdown of circSMAD2 inhibits the tumorigenesis of gallbladder cancer through binding with eIF4A3.
Gallbladder cancer (GBC) is the seventh most common gastrointestinal cancer worldwide. This study aimed to investigate the function of circSMAD2 in GBC. To investigate the function of circSMAD2 in GBC, the level of circSMAD2 in GBC cells was detected by RT-qPCR. CCK-8 assay was performed to investigate the cell viability. Cell apoptosis was tested by flow cytometry. In addition, transwell assay was used to detect the cell migration and invasion. RIP and RNA pull-down were used to explore the relation among circSMAD2, eIF4A3 and SMAD2. Meanwhile, xenograft mice model was established to investigate the function of circSMAD2 in GBC. The data revealed that circSMAD2 was upregulated in GBC, and circSMAD2 knockdown significantly inhibited the viability of GBC cells. In addition, circSMAD2 siRNA notably induced the apoptosis in GBC cells. The migration and invasion of GBC cells were obviously suppressed in the presence of circSMAD2 siRNA. Meanwhile, circSMAD2 suppressed the binding between eukaryotic translation initiation factor 4A3 (eIF4A3) and SMAD2 through binding with eIF4A3. Knockdown of circSMAD2 notably inhibited the expression of SMAD2 in GBC cells, and SMAD2 overexpression partially reversed the anti-tumor effect of circSMAD2 knockdown. Finally, circSMAD2 siRNA significantly inhibited the tumor growth of GBC in vivo. Knockdown of circSMAD2 inhibits the tumorigenesis of gallbladder cancer through binding with eIF4A3. Thus, our study provided a new strategy for the treatment of GBC.


Altered neuropeptide Y Y1 responses in mesenteric arteries in rats with congestive heart failure.
The aim of the present study was to elucidate if the potentiating effect of neuropeptide Y on various vasoactive agents in vitro is (1) altered in mesenteric arteries from rats with congestive heart failure and (2) mediated by the neuropeptide Y Y1 receptor. The direct vascular effects of neuropeptide Y and its modulating effects on the contractions induced by endothelin-1-, noradrenaline-, 5-hydroxytryptamine (5-HT)-, U46619-(9,11-dideoxy-11alpha, 9alpha-epoxymethano-prostaglandin F2alpha) and ATP, and acetylcholine-induced dilatations were studied in the presence and absence of the neuropeptide Y Y1 antagonist, BIBP3226 (BIBP3226¿(R)-N2-(diphenylacetyl)-N-[(4-hydroxyphenyl)methyl ]-D-arginine-amide¿). Neuropeptide Y, per se, had no vasoactive effect in the arteries. The potency of endothelin-1 was significantly decreased in congestive heart failure rats. Neuropeptide Y and neuropeptide Y-(13-36) potentiated the endothelin-1-induced contraction in congestive heart failure mesenteric arteries. In 20% of the congestive heart failure rats, sarafotoxin 6c induced a contraction of 31+/-4%. Neuropeptide Y also potentiated U46619- and noradrenaline-induced contractions but not 5-HT-induced contractions in congestive heart failure arteries. In sham-operated animals neuropeptide Y potentiated noradrenaline- and 5-HT-induced contractions. These potentiations were inhibited by BIBP3226. Acetylcholine induced an equipotent relaxation in both groups which was unaffected by neuropeptide Y. In conclusion, neuropeptide Y responses are altered in congestive heart failure rats. The potentiating effect differs between vasoactive substances. Neuropeptide Y Y1 and non-neuropeptide Y1 receptors are involved.


Unilateral pigmentary degeneration of the retina associated with heterochromia iridis.
For the past 5 years, a 56-year-old patient has been displaying monocular progressive pigmentary changes in the left eye. Heterochromy of the left eye has been known since childhood. The other eye is clinically and functionally normal. The patient was adopted and he has no children. Therefore, we have no family history. The patient was examined clinically and by means of electroretinography, electrooculography, perimetry, computer tomography, pulsatile ocular blood flow (POBF) measurement, serology and Doppler sonography. Electrophysiology displayed a considerable reduction of scotopic and photopic ERGs, a reduced dark-through, and a reduced light-rise in the left eye, whereas the fellow eye was normal. The visual field was limited to 5 deg around the fixation point, and a peripheral crescent-shaped arch encircled the temporal-inferior quadrant concomitant to the pigmentary changes. By computer tomography and Doppler sonography a vascular affection was excluded. The left eye displayed lower POBF values. All serological tests were found negative. The clinical picture and negative exclusion criteria indicate a unilateral retinitis pigmentosa. However, with regard to the literature an unequivocal diagnosis can only be made upon hereditary evidence.


Plastid translation in organello and in vitro during light-induced development in Euglena.
Plastids were isolated from dark-grown Euglena gracilis, from cells exposed to light for 3 h, and from normal, light-grown cells. Plastid protein synthesis was then measured either in organello, by incorporation of [35S]Met into isolated plastids, or in vitro, by programming wheat germ, reticulocyte, or Escherichia coli translation systems with RNA isolated from the plastids. Specific and total rates of protein synthesis in organello increased during light-induced development about 100-fold. In contrast, specific mRNA activity of plastid RNA increased no more than 3-fold, and the estimated total mRNA activity of plastids increased less than 10-fold. Protein synthesis in plastids appears therefore to be subject to strong regulation at a level other than that of transcription superimposed on weak transcriptional regulation. The synthesis of the large subunit of ribulose-bisphosphate carboxylase in particular appears to be under translational control. There is also qualitative evidence for regulation. Most plastid mRNAs, as indicated by translation in vitro, electrophoresis, and fluorography of their translation products, increase with light-induced development as a coordinate set. A second or proplastid set can be detected in RNAs from immature plastids, but not from mature chloroplasts. A few mRNAs appear only later in development, corresponding to a delayed set. Most translation products measured in organello also appear to belong to the coordinate set, a few to the proplastid and delayed sets, and a further few to a transient set, which appears only early in development.


Direct quality control of glycoengineered erythropoietin variants.
Recombinant production of glycoprotein therapeutics like erythropoietin (EPO) in mammalian CHO cells rely on the heterogeneous N-glycosylation capacity of the cell. Recently, approaches for engineering the glycosylation capacities of mammalian cells for custom designed glycoforms have been developed. With these opportunities there is an increasing need for fast, sensitive, and global analysis of the glycoproteoform landscape produced to evaluate homogeneity and consistency. Here we use high-resolution native mass spectrometry to measure the glycoproteoform profile of 24 glycoengineered variants of EPO. Based on the unique mass and intensity profiles of each variant, we classify them according to similarities in glycosylation profiles. The classification distinguishes EPO variants with varying levels of glycan branchingand sialylation, which are crucial parameters in biotherapeutic efficacy. We propose that our methods could be of great benefit in the characterization of other glycosylated biopharmaceuticals, ranging from the initial clonal selection to batch-to-batch controls, and the assessment of similarity between biosimilar/biobetter products.


Design, synthesis and pharmacological evaluation of 2,3-dihydrobenzofuran IRAK4 inhibitors for the treatment of diffuse large B-cell lymphoma.
Interleukin-1 receptor associated kinase 4 (IRAK4) is a critical mediator of MYD88 L265P-induced NF-κB activation, indicating it is a promising therapeutic target for diffuse large B-cell lymphoma (DLBCL). Herein we report the discovery of a series of 2,3-dihydrobenzofuran IRAK4 inhibitors through structure-based drug design. The representative compound 22 exhibited strong IRAK4 inhibitory potency (IRAK4 IC50 = 8.7 nM), favorable kinase selectivity and high antiproliferative activity against the MYD88 L265P DLBCL cell line (OCI-LY10 IC50 = 0.248 μM). Compound 22 also exhibited the ability to inhibit the activation of IRAK4 signaling pathway and induce apoptosis in MYD88 L265P DLBCL cell line. In combination with Bruton's tyrosine kinase (BTK) inhibitor ibrutinib, 22 showed enhanced apoptosis-inducing effect and antiproliferative potency. The most advanced compound 22 in this inhibitor series holds promise for further development into efficacious and selective IRAK4 inhibitors for the treatment of DLBCL.


Physical characteristics of modern radiographic screen-film systems.
The most important characteristics that determine the performance of a screen-film system are system sensitivity (speed), slope or the average gradient of the characteristic curve, latitude of the film, system resolution (MTF) and system noise. In addition to these, one has to consider secondary characteristics such as the base plus fog level, the base tint, safelight sensitivity, reciprocity law failure and film granularity while evaluating performance. A comprehensive study of many of these characteristics has been made for a number of film-screen systems on the market. The data has been tabulated in the form of what are called "characteristic tables of radiographic film-screen systems." In these tables, characteristics such as average gradient, base plus fog level, etc. which depend on film alone, appear along the horizontal axis and MTF values which essentially depend on screens only appear along the vertical axis. The body of the table contains the absolute speeds in units of mR-1 at 80 KVP (heavy filtration) and a quantum mottle index of the various film-screen combinations tested, since these depend both on the film as well as on the screens.


The effects of music therapy on amino acid neurotransmitters: Insights from an animal study.
This study aimed to explore the neurobiological effects of "Chinese Traditional Five-Elements Music Therapy" on rats and to determine its effects on amino acid neurotransmitter levels, the excitatory/inhibitory(E/I) balance and the Glu-Gln cycle. Male Wistar rats were assigned at random to the experimental groups (Gong/powerful; Shang/sad; Jue/gentle; Zhi/joyful; Yu/serene music group) and the control group(n=8/group).The experimental groups were exposed daily to music(2 hours per day; mild sound pressure levels, between 50 and 60 dB) for 28 consecutive days. Finally, we hypothesized concentrations of Glu and GABA to match the music types and measured additionally Asp, Gly, Gln, and Glu/GABA ratio in striatum by UPLC-3QMS. Effects in the predicted direction were observed for Gong (Glu +, GABA -); Shang (Glu -; GABA +); Jue (Glu 0; GABA 0); and Zhi (Glu +; GABA -); but not Yu music (Glu + contrary to hypothesis; GABA + as hypothesized)." In addition, significant difference in Gln levels were also present in the zhi, Gong and Yu music groups compared to the controls. Our study showed that different melodic music produced different effects on amino acid neurotransmitter levels. "Chinese Traditional Five-Elements Music Therapy" affected the amino acid neurotransmitter levels, the E/I balance and the Glu-Gln cycle in the striatum of rats, which may reflect altered glutamatergic and GABAergic system.


Effect of ICRF-187 pretreatment against doxorubicin-induced delayed cardiotoxicity in the rat.
Doxorubicin (DXR), administered iv in rats at the weekly dose of 3 mg/kg for 5 weeks, significantly impaired body weight gain and induced irreversible ECG alterations, mainly consisting of a progressive prolongation of ST and QT intervals. Five weeks after the last DXR administration, the contractile performance of atria isolated from treated animals was significantly reduced. At the same time, relevant morphologic lesions, consisting of myocyte vacuolization and myofibrillar loss, were also present in the myocardium of the same rats. The study showed that ICRF-187, administered ip at a dose of 125 mg/kg, significantly prevented body weight loss. QT and ST prolongation, and the decreased contractile force induced by DXR. In addition, ICRF-187 caused a significant reduction in incidence and severity of myocardial lesions. The cardioprotective effect of ICRF-187 is not mediated by a modification in DXR pharmacokinetics in heart, since the drug was actually found to increase DXR uptake in myocardial cells.


A model system for [NiFe] hydrogenase maturation studies: Purification of an active site-containing hydrogenase large subunit without small subunit.
The large subunit HoxC of the H2-sensing [NiFe] hydrogenase from Ralstonia eutropha was purified without its small subunit. Two forms of HoxC were identified. Both forms contained iron but only substoichiometric amounts of nickel. One form was a homodimer of HoxC whereas the second also contained the Ni-Fe site maturation proteins HypC and HypB. Despite the presence of the Ni-Fe active site in some of the proteins, both forms, which lack the Fe-S clusters normally present in hydrogenases, cannot activate hydrogen. The incomplete insertion of nickel into the Ni-Fe site provides direct evidence that Fe precedes Ni in the course of metal center assembly.


Comparative muscle development of scyphozoan jellyfish with simple and complex life cycles.
Simple life cycles arise from complex life cycles when one or more developmental stages are lost. This raises a fundamental question - how can an intermediate stage, such as a larva, be removed, and development still produce a normal adult? To address this question, we examined the development in several species of pelagiid jellyfish. Most members of Pelagiidae have a complex life cycle with a sessile polyp that gives rise to ephyrae (juvenile medusae); but one species within Pelagiidae, Pelagia noctiluca, spends its whole life in the water column, developing from a larva directly into an ephyra. In many complex life cycles, adult features develop from cell populations that remain quiescent in larvae, and this is known as life cycle compartmentalization and may facilitate the evolution of direct life cycles. A second type of metamorphic processes, known as remodeling, occurs when adult features are formed through modification of already differentiated larval structures. We examined muscle morphology to determine which of these alternatives may be present in Pelagiidae. We first examined the structure and development of polyp and ephyra musculature in Chrysaora quinquecirrha, a close relative of P. noctiluca with a complex life cycle. Using phallotoxin staining and confocal microscopy, we verified that polyps have four to six cord muscles that persist in strobilae and discovered that cord muscles is physically separated from ephyra muscle. When cord muscle is removed from ephyra segments, normal ephyra muscle still develops. This suggests that polyp cord muscle is not necessary for ephyra muscle formation. We also found no evidence of polyp-like muscle in P. noctiluca. In both species, we discovered that ephyra muscle arises de novo in a similar manner, regardless of the life cycle. The separate origins of polyp and ephyra muscle in C. quinquecirrha and the absence of polyp-like muscle in P. noctiluca suggest that polyp muscle is not remodeled to form ephyra muscle in Pelagiidae. Life cycle stages in Scyphozoa may instead be compartmentalized. Because polyp muscle is not directly remodeled, this may have facilitated the loss of the polyp stage in the evolution of P. noctiluca.


Protein synthesis during sleep consolidates cortical plasticity in vivo.
Sleep consolidates experience-dependent brain plasticity, but the precise cellular mechanisms mediating this process are unknown [1]. De novo cortical protein synthesis is one possible mechanism. In support of this hypothesis, sleep is associated with increased brain protein synthesis [2, 3] and transcription of messenger RNAs (mRNAs) involved in protein synthesis regulation [4, 5]. Protein synthesis in turn is critical for memory consolidation and persistent forms of plasticity in vitro and in vivo [6, 7]. However, it is unknown whether cortical protein synthesis in sleep serves similar functions. We investigated the role of protein synthesis in the sleep-dependent consolidation of a classic form of cortical plasticity in vivo (ocular dominance plasticity, ODP; [8, 9]) in the cat visual cortex. We show that intracortical inhibition of mammalian target of rapamycin (mTOR)-dependent protein synthesis during sleep abolishes consolidation but has no effect on plasticity induced during wakefulness. Sleep also promotes phosphorylation of protein synthesis regulators (i.e., 4E-BP1 and eEF2) and the translation (but not transcription) of key plasticity related mRNAs (ARC and BDNF). These findings show that sleep promotes cortical mRNA translation. Interruption of this process has functional consequences, because it abolishes the consolidation of experience in the cortex.


Synthesis, DNA recognition and cleavage studies of novel tetrapeptide complexes, Cu(II)/Zn(II)-Ala-Pro-Ala-Pro.
New tetrapeptide complexes Cu(II)·Ala-Pro-Ala-Pro (1) and Zn(II)·Ala-Pro-Ala-Pro (2) were synthesized from the reaction of tetrapeptide, Ala-Pro-Ala-Pro and CuCl2/ZnCl2 and were thoroughly characterized by elemental analysis, IR,(1)H and (13)C NMR (in case of 2), ESI-MS, UV and molar conductance measurements. The solution stability study was carried out employing UV-vis absorption titrations over a broad range of pH which suggested the stability of the complexes in solution. In vitro interaction of complexes 1 and 2 with CT-DNA was studied employing UV-vis, fluorescence, circular dichroic and viscometry studies. To throw insight into molecular binding event at the target site, UV-vis titrations of 1 and 2 with mononucleotides of interest viz.; 5'-GMP and 5'-TMP were carried out. Cleavage activity of the complexes with pBR322 plasmid DNA was evaluated by agarose gel electrophoresis and, the electrophoresis pattern demonstrated that both the complexes 1 and 2 are efficient cleavage agents. Further, the Cu(II) complex displayed efficient oxidative cleavage of supercoiled DNA while various reactive oxygen species are responsible for the cleavage in Zn(II) complex.


Circulating 25-hydroxyvitamin D levels and hypertension risk after adjusting for publication bias.
Previous systematic reviews reported that serum vitamin D deficiency was associated with risk of hypertension. The aim was to conduct a meta-epidemiological analysis for evaluating the potential effects of publication bias. The selection criterion was defined as a follow-up study for evaluating the association between circulating 25-hydroxyvitam D level and hypertension risk in adults. A funnel plot and Egger's test were used to detect a publication bias. If a publication bias was identified, trim-and-fill analysis (TFA) with linear estimator was performed to estimate a summary relative risk (sRR). The meta-analysis of 13 cohorts resulted in the lower the vitamin D, the higher the risk of hypertension statistically significant (sRR, 1.22; 95% confidence interval [CI], 1.05 to 1.41). But The P-value of Egger's test (=0.015) and asymmetry of the funnel plot showed that there was a publication bias. TFA resulted in that statistical significance disappeared in the association between vitamin D level and hypertension risk in total cohorts (filled sRR, 1.03; 95% CI, 0.89 to 1.18) as well as men and women cohorts. The publication bias-adjusted results by TFA had no statistically significant association between vitamin D levels and the risk of hypertension. The significant results in previous systematic reviews might be interpreted as due to publication bias.


Sequential monitoring of covariate adaptive randomized clinical trials with sample size re-estimation.
Once scientific questions are determined, other design features of clinical trials including increasing the power while controlling the type I error rate, planning interim analysis, and achieving treatment balance among subgroups will be either required or preferred. We propose to sequentially monitor the covariate adaptive randomization (CAR) procedures with sample size re-estimation (SSR) to satisfy a variety of design objectives of clinical trials. However, each of the three adaptive designs (sequential monitoring, CAR, and SSR) poses a challenge to the control of the type I error rate. In this paper, we investigated how to utilize the advantages of the three adaptive methods and control the type I error rate. We proved that the asymptotic joint distribution of the sequential statistics follows the asymptotic canonical joint distribution defined in Jennison and Turnbull [14]. Besides, numerical studies demonstrated that our methods could control the type I error rate, increase the power, and lead to much-improved treatment balance across subgroups.


Influence of goals on modular brain network organization during working memory.
Top-down control underlies our ability to attend relevant stimuli while ignoring irrelevant, distracting stimuli and is a critical process for prioritizing information in working memory (WM). Prior work has demonstrated that top-down biasing signals modulate sensory-selective cortical areas during WM, and that the large-scale organization of the brain reconfigures due to WM demands alone; however, it is not yet understood how brain networks reconfigure between the processing of relevant versus irrelevant information in the service of WM. Here, we investigated the effects of task goals on brain network organization while participants performed a WM task that required participants to detect repetitions (e.g., 0-back or 1-back) and had varying levels of visual interference (e.g., distracting, irrelevant stimuli). We quantified changes in network modularity-a measure of brain sub-network segregation-that occurred depending on overall WM task difficulty as well as trial-level task goals for each stimulus during the task conditions (e.g., relevant or irrelevant). First, we replicated prior work and found that whole-brain modularity was lower during the more demanding WM task conditions compared to a baseline condition. Further, during the WM conditions with varying task goals, brain modularity was selectively lower during goal-directed processing of task-relevant stimuli to be remembered for WM performance compared to processing of distracting, irrelevant stimuli. Follow-up analyses indicated that this effect of task goals was most pronounced in default mode and visual sub-networks. Finally, we examined the behavioral relevance of these changes in modularity and found that individuals with lower modularity for relevant trials had faster WM task performance. These results suggest that brain networks can dynamically reconfigure to adopt a more integrated organization with greater communication between sub-networks that supports the goal-directed processing of relevant information and guides WM.


Enzymatic distinction between two subgroups of autosomal recessive lamellar ichthyosis.
It has been proposed that the autosomal recessive lamellar ichthyoses may be divided into two subgroups, the erythrodermic (EARLI) and non-erythrodermic (NEARLI) forms. We report measurements of the enzymes beta-glucosidase, a recently described phosholipase, a short-chain carboxylesterase ("butyrase"), and a long-chain carboxylesterase ("palmitase") in aqueous extracts of scales from patients diagnosed according to clinical and micromorphologic criteria, and show that beta-glucosidase and phospholipase tend to be lower in the EARLI group, whereas butyrase is relatively low in the NEARLI group. The internal ratio of either butyrase/glucosidase or butyrase/phospholipase yields a clear separation of the two subgroups, supporting the concept of heterogeneity in this group of diseases.


Maternal cigarette smoking before and during pregnancy and the risk of preterm birth: A dose-response analysis of 25 million mother-infant pairs.
Most of the women who smoke before pregnancy continue smoking during pregnancy, and some start to quit smoking after being pregnant, although existing guidelines for pregnancy recommend that women who smoke should quit smoking before pregnancy. Findings about the timing and intensity of maternal smoking, especially low-intensity smoking (1-9 cigarettes per day), and preterm birth are still inconsistent and ambiguous. This study aimed to examine the association of the timing of smoking and doses of smoking before pregnancy and during the first or second trimester of pregnancy with preterm birth in a large-scale population-based retrospective cohort study. We used nationwide birth certificate data from singleton mother-infant pairs in the United States National Vital Statistics System, 2011-2018. All adult women with live singleton births, without preexisting hypertension or diabetes, and with complete data on smoking and gestational age at delivery were included. Participants reported their smoking status (yes or no) and daily number of cigarettes consumed before and during each trimester of pregnancy. The outcome of interest was preterm birth, defined as a birth before 37 weeks of gestation. Logistic regression models were used to estimate the odds ratio (OR) with 95% confidence intervals (CIs) of preterm birth associated with smoking status and the number of cigarettes consumed, adjusting for maternal age, race/ethnicity, parity, education levels, prepregnancy BMI, previous history of preterm birth, marital status, infant sex, and initiation of prenatal care. This study included 25,623,479 women, with a mean age of 29 years (range 20-50 years); 13,742,486 (53.6%) participants were of non-Hispanic white ancestry, 5,971,598 (23.3%) of Hispanic ancestry, and 3,417,456 (13.34%) of non-Hispanic black ancestry. The prevalence of preterm birth was 9.3% (n = 2,378,398). We found that maternal smoking during pregnancy, even at a very low level of intensity, was associated with an increased risk of preterm delivery. The adjusted ORs (95% CI) of preterm birth for mothers who smoked 1-2, 3-5, 6-9, 10-19, and ≥20 cigarettes per day during the first trimester compared with mothers who did not smoke were 1.31 (1.29-1.33), 1.31 (1.30-1.32), 1.33 (1.31-1.35), 1.44 (1.43-1.45), and 1.53 (1.52-1.55), respectively (all P values < 0.001), whereas for those who smoked during the second trimester, the corresponding ORs were 1.37 (1.35-1.39), 1.36 (1.35-1.38), 1.36 (1.34-1.38), 1.48 (1.47-1.49), and 1.59 (1.58-1.61), respectively (all P values < 0.001). Furthermore, smokers who quit before pregnancy, regardless of smoking intensity, had a comparable risk of preterm birth with nonsmokers, although this was not the case when cessation occurred in the first or second trimester of pregnancy. The major limitation of this study is the self-reported information about smoking, which may be subject to information bias. In addition, we cannot rule out the possibility of residual confounding caused by unmeasured factors in an observational research design. In this study, we observed that low-intensity cigarette consumption during either the first or second trimester of pregnancy, even as low as 1-2 cigarettes per day, was associated with an increased risk of preterm birth. These findings suggest that there is no safe level or safe trimester for maternal smoking during pregnancy. Women of reproductive age who smoke should be strongly encouraged and supported to quit smoking before pregnancy.


Contralateral voluntary hand movement inhibits human parkinsonian tremor and variably influences essential tremor.
While voluntary movement blocks Parkinsonian rest tremor (PT), essential tremor (ET) is enforced by postural and/or kinetic action. We studied the effect of contralateral externally- and internally triggered hand movement on PT and ET to investigate the transhemispheric influences on tremor genesis. We measured the changes of tremor peak frequency power after flash signal (F), flash triggered (FM) and self-paced (SPM) movement of the contralateral hand in nine PT and seven ET patients using accelerometer. PT significantly decreased both during FM and SPM tasks, suggesting that it is generated by a constant subcortico-cortical network, which includes higher order motor areas. Intensity of ET showed a remarkable intra- and interindividual variability both during FM and SPM reflecting a different generator circuitry with variable functional connections.


Insect control in socialist China and the corporate United States: the act of comparison, the tendency to forget, and the construction of difference in 1970s U.S.-Chinese scientific exchange.
In 1975, a delegation of U.S. entomologists traveled to socialist China to observe Chinese insect control science. Their overwhelmingly positive reports highlighted in relief the pernicious effects of pesticide corporations on U.S. agriculture; some entomologists hoped this would goad the United States to catch up to China in environmentally sensible insect control practices. Of course, insect control in socialist China carried its own political baggage, some of which-for example, mass mobilization and self-reliance--the state made highly visible to visitors, and some of which--for example, harsh treatment of scientists--it sought to obscure. For both the U.S. and the Chinese participants, the act of comparison itself was of primary significance in the exchange, allowing them to construct socialist Chinese science as refreshingly different from U.S. science. At the same time, however, this construction of difference meant forgetting the much longer transnational history in which U.S. and Chinese entomology had been intertwined.


Preparation and characterization of a novel variant of human tumor necrosis factor-related apoptosis-inducing ligand from the rhesus monkey, Macaca mulatta.
Human tumor necrosis factor-related apoptosis-inducing ligand (hTRAIL) and its variants are attractive antitumor drug candidates. The predicted amino acid sequence of the functional extracellular domain of Macaca mulatta TRAIL (mmTRAIL) was found to differ from that of hTRAIL at four positions. In this study, the gene encoding mmTRAIL was cloned and recombinantly expressed in Escherichia coli at a yield of approximately 20-30 mg/L, which was two times higher than that of hTRAIL. SDS-PAGE showed that denatured mmTRAIL and hTRAIL had similar molecular weights. However, size-exclusion chromatography and dynamic light scattering (DLS) analysis demonstrated that the molecular size of native mmTRAIL was smaller than that of native hTRAIL. Cooling solutions of these proteins from room temperature to 0 °C induced considerable precipitation of hTRAIL but not of mmTRAIL, indicating that mmTRAIL was more soluble than hTRAIL at low temperatures. Additionally, mmTRAIL was more resistant than hTRAIL to N-bromosuccinimide (NBS)-induced precipitation. Although mmTRAIL and hTRAIL showed comparable nanomolar affinities for human death receptors, the dissociation rate of the mmTRAIL-receptor complex was slower than that of the hTRAIL-receptor complex, suggesting that the mmTRAIL-receptor complex was more stable. Moreover, mmTRAIL induced caspase-dependent apoptosis in human tumor cells with an IC50 that was two to three times lower than that of hTRAIL. However, in vivo evaluation demonstrated that mmTRAIL or hTRAIL led to a similar level of tumor suppression in mice bearing COLO205 xenografts. Nevertheless, the advantage of its better solubility should promote the production and further use of mmTRAIL in cancer biotherapy.


Translation of evidence-based guidelines in preventive cardiology: the EUROACTION program.
The report from the EUROACTION Study Group demonstrates that a nurse-coordinated, multidisciplinary, family-based program results in improvements in patients' lifestyle, control of blood pressure, and use of cardioprotective drugs, when compared with standard care. The results of the study support the need for organized and systematic approaches to individualized cardiovascular risk reduction in patients with, or at high risk for, coronary heart disease. The EUROACTION program is an example of effectiveness research, which aims to develop and validate strategies to implement interventions previously shown to be efficacious.


Impact of hemodialysis on the abnormal glucose and alanine kinetics of chronic azotemia.
Rates of alanine and glucose turnover and precursor-product interrelationships were determined in patients on chronic hemodialysis and in matched controls using simultaneous primed injection-continuous infusions of [U-14C] alanine and [2-3h] glucose. In eight chronically dialyzed patients studied before their first dialysis of the week, glucose turnover was 866 +/- 120 micromole/min (mean +/- SE); after their last dialysis of the week, glucose turnover was 880 +/- 63 micromole/min. These rates were 35% (p less than 0.05) and 37% (p less than 0.01) greater than rates observed in ten normal volunteers (642 +/- 28.3 micromole/min). Fasting glucose and insulin levels in dialyzing patients were unchanged from normal. Alanine turnover was increased predialysis (318 +/- 55.2 micromoles/min; p less than 0.01) and postdialysis (248 +/- 32.4 micromole/min; p less than 0.01) as compared to normal (168 +/- 14.3 micromole/min). In patients pre- and postdialysis, gluconeogenesis from alanine was increased to 34.6 +/- 10.9 micromole/min (p less than 0.05) and 39.0 +/- 6.33 micromole/min (p less than 0.05) compared to 20.9 +/- 1.63 micromole/min in normal subjects. We conclude that neither acute nor chronic hemodialysis corrects the increased glucose and alanine production and utilization and gluconeogenesis observed in chronic renal failure.


A case of Aspergillus myocarditis associated with septic shock.
We report a case of isolated myocardial involvement due to Aspergillus sp., in a patient with multiple organ failure but no demonstrable immune deficiency apart from corticosteroid therapy given for the three weeks prior to hospitalization. This patient died of septic shock 7 days after hospitalization. Aspergillus was isolated only once from a sputum specimen obtained 24 h before death. At post-mortem examination there are emboli and myocardial lesions containing clusters of hyphae as Aspergillus confirmed by indirect immunofluorescence. The other organs were free of fungi. This observation is remarkable because the cardiac involvement was isolated and no immunodeficiency was present.


Ursane-type pentacyclic triterpenoids as useful platforms to discover anticancer drugs.
This review highlights the potential of natural and semisynthetic ursane-type triterpenoids as candidates for the design of multi-target bioactive compounds, with focus on their anticancer effects. A brief illustration of the biosynthesis, sources, and general biological effects of the main classes of naturally occurring pentacyclic triterpenoids (PTs) are provided.


Chronic CNS-mediated cardiometabolic actions of leptin: potential role of sex differences.
Previous studies using male rodents showed the adipocyte-derived hormone leptin acts in the brain to regulate cardiovascular function, energy balance, and glucose homeostasis. The importance of sex differences in cardiometabolic responses to leptin, however, is still unclear. We examined potential sex differences in leptin's chronic central nervous system (CNS)-mediated actions on blood pressure (BP), heart rate (HR), appetite, and glucose homeostasis in normal and type 1 diabetic rats. Female and male Sprague-Dawley (SD) rats were instrumented with intracerebroventricular cannulas for continuous 7-day leptin infusion (15 µg/day), and BP and HR were measured by telemetry 24 h/day. At baseline, females had lower mean arterial pressure (MAP) (96 ± 3 vs. 104 ± 4 mmHg, P < 0.05) but higher HR (375 ± 5 vs. 335 ± 5 beats/min, P < 0.05) compared with males. After leptin treatment, we observed similar increases in BP (∼3 mmHg) and HR (∼25 beats/min) in both sexes. Females had significantly lower body weight (BW, 283 ± 2 vs. 417 ± 7 g, P < 0.05) and caloric intake (162 ± 20 vs. 192 ± 9 kcal/kg of body wt, P < 0.05) compared with males, and leptin infusion reduced BW (-10%) and caloric intake (-62%) similarly in both sexes. In rats with streptozotocin-induced diabetes (n = 5/sex), intracerebroventricular leptin treatment for 7 days completely normalized glucose levels. The same dose of leptin administered intraperitoneally did not alter MAP, HR, glucose levels, or caloric intake in normal or diabetic rats. These results show that leptin's CNS effects on BP, HR, glucose regulation, and energy homeostasis are similar in male and female rats. Therefore, our results provide no evidence for sex differences in leptin's brain-mediated cardiovascular or metabolic actions.


Incomplete Pentalogy of Cantrell--A Case Report.
Cantrell's Pentalogy is a rare congenital malformation consists of supraumbilical abdominal wall defect, defect in the lower part of sternum, agenesis of anterior portion of diaphragm, an absence of the diaphragmatic part of the pericardium and intracardiac malformation. This case report presents a female neonate, who was born at 40 weeks of gestation weighing 2400 gm and was admitted 4 hours after delivery with the complaints of something coming out from chest. On physical examination her vital signs were within normal limit, she had a systolic murmur on heart at lower left sternal area and there was a vascular structure present on the upper part of abdomen which was pulsatile and pulsation was synchronized with cardiac pulsation. On investigation chest X-ray lateral view showed absence of lower part of sternum, echocardiography findings were different in different institutes. Echocardiography findings at Mymensingh medical college hospital (MMCH) were large VSD (Ventriculo septal defect), ASD (Atrial septal defect) and rotated heart. On colour Doppler ultrasonogram at MMCH showed there were VSD, ASD, and a small epigastric swelling which was vascular and appears to be attached to the apex of the heart. On the other hand echocardiography findings of LAB AID hospital in Dhaka were Dextrocardia, complete AV (atrio ventricular) canal defect, almost common atrium, almost single ventricle, common AV valve, double outlet right ventricle (DORV), Cortriatriatum, mild A-V valve regurgitation and severe pulmonary hypertension (PAH). Echocardiography was also done at national heart foundation hospital in Dhaka. Findings were situs solitus, mesocardia to dextrocardia, DORV, large VSD, mild mitral inflow, mild TR (triuspid regurgitation), good LV (left ventricle) and RV (right ventricle) systolic function. Cardiologists at Dhaka in Bangladesh were suggested for surgery. Then the patient was consulted at Naryan Institute of Cardiac Science, Chennai in India. Here echocardiography findings were dextroversion/dextrocardia, DILV (Double inlet left ventricle), large inlet VSD with bidirectional shunt, mild TR, severe PAH with good ventricular function. Cardiologists in India were given comment about this patient. This patient was highly risky for surgery. They advised medical treatment and requested to review after one year. By taking medical treatment patient condition is well except failure to thrive and cyanosis develops during feeding and crying according to the statement of guardian of the patient. This case has 3 criterias among the five criteria of Cantrell's Pentalogy. So, it is incomplete Pentalogy of Cantrell.


Suboptimal glycemic control in adolescents and young adults with type 1 diabetes from 2011 to 2020 across Australia and New Zealand: Data from the Australasian Diabetes Data Network registry.
Competing challenges in adolescence and young adulthood can distract from optimal type 1 diabetes (T1D) self-management, and increase risks of premature morbidity and mortality. There are limited data mapping the glycemic control of people with T1D in this age group, across Australasia. Clinical data were extracted from the Australasian Diabetes Data Network, a prospective clinical diabetes registry. Inclusion criteria were individuals with T1D aged 16-25 years at their last recorded T1D healthcare visit (from 1st January 2011 to 31st December 2020), with T1D duration of at least 1 year. Data were stratified by two last recorded T1D healthcare visit ranges, while generalized estimated equation (GEE) modeling was used to examine factors associated with HbA1c across visits during the 10 year period. Data from 6329 young people (52.6% male) attending 24 diabetes centers across Australasia were included. At the last visit within the most recent 5 years, mean ± SD age was 18.5 ± 2.3 years, T1D duration was 8.8 ± 4.7 years and HbA1c was 8.8 ± 1.8% (72.2 ± 19.9 mmol/mol); only 12.3% had an HbA1c below the international target of <7.0% (53 mmol/mol). Across all T1D healthcare visits, in GEE modeling, higher HbA1c was associated with female sex (B = 0.20; 95% CI 0.12 to 0.29, p < 0.001), longer T1D duration (B = 0.04, 0.03 to 0.05, p < 0.001). Lower HbA1c was associated with attendance at a pediatric T1D healthcare setting (B = -0.33, -0.45 to -0.21, p < 0.001) and use of CSII versus BD/MDI therapy (B = -0.49, -0.59 to 0.40, p < 0.001). This Australasian study demonstrates widespread and persistent sub-optimal glycemic control in young people with T1D, highlighting the urgent need to better understand how healthcare services can support improved glycemic control in this population.


Peritoneal dialysis catheter placement, outcomes and complications.
Peritoneal dialysis (PD) is a commonly used method for renal support in pediatric patients and can be associated with the risk of post-surgical complications. We evaluated method of placement of PD catheters with regard to post-surgical complications. PD catheters placed at two institutions between 2005 and 2017 were reviewed. Complication rates were evaluated based on method of placement, delayed usage, omentectomy, and patient age using Fisher's exact test, two-sided, with significance set at 0.05. Factors influencing complication were evaluated with multivariate logistic regression and Kaplan-Meier survival analysis. There were 130 patients with 157 catheters placed, ranging in age from 1 day to 23 years. There was no significant difference in complication rate by method of placement or delayed usage. Infants were significantly more likely to experience leakage (21% vs 8%, p 0.036) and hernias (15% vs 5%, p 0.030). Patients that underwent an omentectomy were less likely to require a catheter replacement (7% vs 27%, p 0.004), and the catheters had a significantly higher survival rate (p 0.009). We found that laparoscopic intervention resulted in catheter salvage. Lateral exit sites may be a risk factor for catheter migration in some patients. Omentectomy is associated with longer PD catheter survival. Laparoscopic salvage of dysfunctional catheters may be a valuable adjunct in management.


Postharvest biocontrol of Colletotrichum gloeosporioides on mango using the marine bacterium Stenotrophomonas rhizophila and its possible mechanisms of action.
The marine bacterium Stenotrophomonas rhizophila was assessed in vitro and in vivo as biocontrol agent against anthracnose disease of mango fruit caused by Colletotrichum gloeosporioides. The results showed that in vitro inhibition of the colony diameter and spore germination of the phytopathogen was due to the production of VOCs, competition for nutrients, and lytic enzymes. When a concentration of 1 × 108 cells ml-1 of the antagonist bacterium was applied to the fruit, disease incidence was reduced by 95%, and the lesion diameter of anthracnose decreased by 85%, which offered greater protection than the synthetic fungicide. This is the first report of antagonistic mechanisms of the marine bacterium S. rhizophila against anthracnose disease in mango, which in this study was found to be more effective than the synthetic fungicide.


Treatment of heterotopic ossification of the elbow following burn injury: recommendations for surgical excision and perioperative prophylaxis using radiation therapy.
Heterotopic ossification (HO) is reported to occur in 0.1-3.3% of elbows after a severe burn, and can significantly limit elbow motion and upper extremity function. The study included 9 patients (11 elbows) treated by the senior author (TW). The surgical technique consisted of making multiple small surgical approaches to remove heterotopic ossification (without raising cutaneous flaps) and concomitantly releasing the elbow capsule and skin contracture. Perioperative radiation therapy was performed to decrease heterotopic ossification recurrence. Outcome measures included postoperative elbow range of motion and Mayo Elbow Performance Score. The average amount of body surface area burned was 54% (range, 10-86%) and mean time from injury to elbow surgery was 416 days (range, 175-860). All elbows had some degree of direct involvement with the thermal injury. Preoperative arc of motion averaged 39° in flexion/extension and 78° in supination/pronation. Four elbows had complete ankylosis in the flexion/extension plane and 1 had only 5° of motion. At last follow-up, arc of motion in flexion/extension averaged 116° and 139° in supination/pronation, an improvement of 77° and 61°, respectively. One recurrence of HO required re-excision. We recommend this multiple-approach surgical technique for treatment of heterotopic ossification and elbow contracture after burn injury, along with perioperative radiation therapy to decrease recurrence. Our surgical approach and treatment resulted in significant gains in elbow motion and upper extremity function with few complications.


Retinal nerve fiber layer thickness in the fellow eyes of normal-tension glaucoma patients with unilateral visual field defect.
To quantitatively evaluate retinal nerve fiber layer (RNFL) thickness in the fellow eyes of normal-tension glaucoma (NTG) patients with unilateral visual field defect. Observational case-control study. Twenty-nine NTG patients with unilateral visual field defect were enrolled in this study. All 29 fellow eyes showed normal visual field. Thirty-one normal eyes of 31 subjects served as controls. The RNFL thickness around the optic disk was determined using Fast RNFL thickness (3.4) of optical coherence tomography. Average and segmental (4 quadrants and 12 clock- hours) RNFL thickness measurements were compared among the three groups. RNFL thicknesses were significantly different among the three groups in the average, superior quadrant (11 and 12 clock-hour segments), and inferior quadrant (6 clock-hour segment) (P = .00, one-way ANOVA and Tukey's tests). RNFL thickness reductions are already present in the fellow eyes of NTG patients with unilateral visual field defect.


Treatment of opioid overdose: current approaches and recent advances.
The USA has recently entered the third decade of the opioid epidemic. Opioid overdose deaths reached a new record of over 74,000 in a 12-month period ending April 2021. Naloxone is the primary opioid overdose reversal agent, but concern has been raised that naloxone is not efficacious against the pervasive illicit high potency opioids (i.e., fentanyl and fentanyl analogs). This narrative review provides a brief overview of naloxone, including its history and pharmacology, and the evidence regarding naloxone efficacy against fentanyl and fentanyl analogs. We also highlight current advances in overdose treatments and technologies that have been tested in humans. The argument that naloxone is not efficacious against fentanyl and fentanyl analogs rests on case studies, retrospective analyses of community outbreaks, pharmacokinetics, and pharmacodynamics. No well-controlled studies have been conducted to test this argument, and the current literature provides limited evidence to suggest that naloxone is ineffective against fentanyl or fentanyl analog overdose. Rather a central concern for treating fentanyl/fentanyl analog overdose is the rapidity of overdose onset and the narrow window for treatment. It is also difficult to determine if other non-opioid substances are contributing to a drug overdose, for which naloxone is not an effective treatment. Alternative pharmacological approaches that are currently being studied in humans include other opioid receptor antagonists (e.g., nalmefene), respiratory stimulants, and buprenorphine. None of these approaches target polysubstance overdose and only one novel approach (a wearable naloxone delivery device) would address the narrow treatment window.


Eight-year results after aortic valve replacement with the CryoLife-O'Brien Stentless Aortic Porcine Bioprosthesis.
The long-term durability and hemodynamics of stentless valves are largely unknown. In this study we prospectively analyzed 8-year clinical results with the CryoLife-O'Brien Stentless Aortic Porcine Bioprosthesis (CryoLife Inc, Kennesaw, Ga) and assessed its hemodynamic performance by serial echocardiography. A total of 206 patients with a mean age of 72.8 years were followed up prospectively after aortic valve replacement with the CryoLife-O'Brien stentless bioprosthesis. Patients have been followed up from 2 to 96 months for mean 56 months. Echocardiography was performed by a single echocardiographer preoperatively, intraoperatively, postoperatively at discharge, 3 to 6 months later, and annually thereafter. The 30-day operative mortality was 4.8%. Sixty-five percent of patients received a valve 25 mm in diameter or larger, and 37% underwent concomitant coronary bypass grafting. Twelve late deaths, none valve-related, have occurred. Severe aortic insufficiency caused by oversizing led to early reoperation in 3 patients. The peak and mean systolic gradients decreased significantly during the first 12 months after implantation (P < .001), and the effective valve areas increased significantly during this interval (P < .001). At 8 years, 2 patients have mild to moderate aortic insufficiency. The actuarial survival at 8 years was 82% +/- 3%. The freedom from endocarditis was 100%, and the freedom from thromboembolic events was 93%. Despite more demanding surgical technique than with conventional bioprostheses, the CryoLife-O'Brien bioprosthesis can be implanted safely in a population predominantly older than 70 years at the time of the operation, with excellent measures of hemodynamics, clinical outcomes, prosthesis durability, and survival through 8 years.


A-type potassium current in myenteric neurons from guinea-pig small intestine.
Biophysical properties of A-type K(+) currents (I(A)) in myenteric neurons from guinea-pig small intestine were studied. I(A) was present in both AH- and S-type myenteric neurons. Reduction of external Ca(2+) did not affect the current. Current density was 13.5+/-10.2 pA/pF in 68 AH-type neurons and 23.4+/-8.2 pA/pF in 31 S-type neurons. S-type neurons appeared to be a homogeneous group based on density of I(A). AH-type neurons were subdivided into two groups with current densities of 9.4+/-4.3 and 25.4+/-4.3 pA/pF. All other biophysical properties of the current were not statistically different for AH- and S-type neurons. Steady-state activation and inactivation curves showed half-activation potentials at -7 mV (k=15. 0 mV) and -86 mV (k=11.5 mV). The curves overlapped at potentials near the resting potential of approximately -55 mV. Time constants for activation ranged from 3.6 to 0.52 ms at test potentials between -20 and 50 mV. Inactivation time constants fell between 41.5 and 11 ms at test potentials between -20 and 50 mV. Time constants for recovery from inactivation fit a double-exponential curve with fast and slow recovery times of 11 and 550 ms. 4-Aminopyridine suppressed I(A) when it was activated at -20 mV following a pre-pulse to -110 mV. Addition of Zn(2+) in the external solution resulted in a concentration-dependent shift of the activation and inactivation curves in the depolarized direction. Zn(2+) slowed the activation and inactivation kinetics of I(A) by factors of 3.3- and 1.2-fold over a wide range of potentials. Elevation of external H(+) suppressed the effect of Zn(2+) with a pK of 7.3-7.4. The effects of Zn(2+) were interpreted as not being due to surface charge screening, because the affinity of Zn(2+) for its binding site on the A-channel was estimated to be between 170 and 312 microM, while the background concentration of Mg(2+) was 10 mM. The enteric nervous system is perceived as an independent integrative nervous system (brain-in-the-gut) that is responsible for local organizational control of motility and secretory patterns of gut behavior. AH- and S-type neurons are synaptically interconnected to form the microcircuits of the enteric nervous system. The results suggest that I(A) is a significant determinant of neuronal excitability for both the firing of nerve impulses and the various synaptic events in the two types of neurons.


Fusion with type 2 macrophages induces melanoma cell heterogeneity that potentiates immunological escape from cytotoxic T lymphocytes.
Evasion from immunity is a major obstacle to the achievement of successful cancer immunotherapy. Hybrids derived from cell-cell fusion are theoretically associated with tumor heterogeneity and progression by conferring novel properties on tumor cells, including drug resistance and metastatic capacity; however, their impact on immune evasion remains unknown. Here, we investigated the potency of tumor-macrophage hybrids in immune evasion. Hybrids were established by co-culture of a melanoma cell line (A375 cells) and type 2 macrophages. The hybrids showed greater migration ability and greater tumorigenicity than the parental melanoma cells. The hybrids showed heterogeneous sensitivity to New York esophageal squamous cell carcinoma-1 (NY-ESO-1)-specific T-cell receptor-transduced T (TCR-T) cells and two out of four hybrid clones showed less sensitivity to TCR-T compared with the parental cells. An in vitro tumor heterogeneity model revealed that the TCR-T cells preferentially killed the parental cells compared with the hybrids and the survival rate of the hybrids was higher than that of the parental cells, indicating that the hybrids evade killing by TCR-T cells efficiently. Analysis of a single-cell RNA sequencing dataset of patients with melanoma revealed that a few macrophages expressed RNA encoding melanoma differentiation antigens including melan A, tyrosinase, and premelanosome protein, which indicated the presence of hybrids in primary melanoma. In addition, the number of potential hybrids was correlated with a poorer response to immune checkpoint blockade. These results provide evidence that melanoma-macrophage fusion has a role in tumor heterogeneity and immune evasion. © 2023 The Pathological Society of Great Britain and Ireland.


[Comparison of maternal and perinatal outcomes in the conservative treatment preterm premature membrane rupture between the use of erythromycin and clindamycin].
premature rupture of membranes occurs between 5 and 15% of pregnancies, of these, 10% occurs at term and preterm 2 to 3.5%. To compare maternal and perinatal outcomes from the use of erythromycin or clindamycin in women with preterm premature rupture of membranes with conservative treatment at the Regional General Hospital No. 36. comparative, prospective, randomized study conducted at the Regional General Hospital No. 36 of the Instituto Mexicano del Seguro Social, Puebla. The most common risk factors were cervical infections (55%) and urinary tract infection (55%). A history of premature rupture of membranes in pregnancy was reported in 12.5% of patients. Group A was prescribed erythromycin and group B, standard-dose clindamycin, these were the independent variables. Maternal outcomes (deciduoendometritis and chorioamnionitis) and perinatal (respiratory distress syndrome, necrotizing enterocolitis and sepsis) were the dependent variables. Fisher exact test was applied to the results of the study. about perinatal outcome, sepsis was expressed more frequently in the clindamycin group (60%) compared with erythromycin (35%). The occurrence of respiratory distress syndrome was similar in both groups, 70 and 75% respectively. For necrotizing enterocolitis, 25 and 5%. Maternal alterations as chorioamnionitis occurred in 20% of patients in group A and 5% in group B. Endometritis results were similar in both groups. comparing the maternal and perinatal outcomes with conservative management of premature rupture of membranes, results were better in the group treated with erythromycin. It is not possible to prove it statistically because of the sample size.


Medicinal benefits of marine invertebrates: sources for discovering natural drug candidates.
Marine invertebrates are one of the major groups of organisms, which could be diversified under the major taxonomic groups of Porifera, Cnidaria, Mollusca, Arthropoda, Echinodermata, and many other minor phyla. To date, range of medicinal benefits and a significant number of marine natural products (MNPs) have been discovered from marine invertebrates. Seafood diet from edible marine invertebrates such as mollusks and crustaceans has been linked with various medicinal benefits to improve human health. Among marine invertebrates, spongers from phylum Porifera is the most dominant group responsible for discovering large number of MNPs, which have been used as template to develop therapeutic drugs. MNPs isolated from invertebrates have shown wide range of therapeutic properties including antimicrobial, antioxidant, antihypertensive, anticoagulant, anticancer, anti-inflammatory, wound healing and immune modulator, and other medicinal effects. Therefore, marine invertebrates are rich sources of chemical diversity and health benefits for developing drug candidates, cosmetics, nutritional supplements, and molecular probes that can be supported to increase the healthy life span of human.


Hahb-4, a sunflower homeobox-leucine zipper gene, is a developmental regulator and confers drought tolerance to Arabidopsis thaliana plants.
Homeodomain-leucine zipper proteins constitute a family of transcription factors found only in plants. Hahb-4 is a member of Helianthus annuus (sunflower) subfamily I. It is regulated at the transcriptional level by water availability and abscisic acid. In order to establish if this gene plays a functional role in drought responses, transgenic Arabidopsis thaliana plants that overexpress Hahb-4 under the control of the 35S Cauliflower Mosaic Virus promoter were obtained. Transformed plants show a specific phenotype: they develop shorter stems and internodes, rounder leaves and more compact inflorescences than their non-transformed counterparts. Shorter stems and internodes are due to a lower rate in cell elongation rather than to a stop in cell division. Transgenic plants were more tolerant to water stress conditions, showing improved development, a healthier appearance and higher survival rates than wild-type plants. Indeed, either under normal or drought conditions, they produce approximately the same seed weight per plant as wild-type plants under normal growth conditions. Plants transformed with a construct that bears the Hahb-4 promoter fused to gusA show reporter gene expression in defined cell-types and developmental stages and are induced by drought and abscisic acid. Since Hahb-4 is a transcription factor, we propose that it may participate in the regulation of the expression of genes involved in developmental responses of plants to desiccation.


Feasibility assessment of patient-controlled EEG home-monitoring: More results from the HOMEONE study.
The feasibility phase of the HOME (Home-Monitoing and Education) project aims to show the practical feasibility of Electroencephalography (EEG)home-monitoring using a patient-controlled mobile system. Its objective is to assess the potential diagnostic and therapeutic yields of home-monitoring compared to conventional healthcare. 16 office-based practitioners chose 97 patients and recorded standard 20-minute EEGs using conventional recorders. After training, the same patients used a patient-controlled mobile dry electrode EEG system in their home environment. The practitioners in charge and two additional raters assessed all recordings. We conducted inter-rater and intra-rater comparisons between the diagnostic findings. 89 patients successfully conducted home-monitoring recordings. The intra-rater comparison results for the diagnostic findings of the conventional recordings and the patient-made recordings show a fair Cohen's kappa value (0.21). Additionally, we documented a change of patient management in 9 cases. The feasibility of EEG home-monitoring using a patient-controlled device is confirmed. The yield of EEG home-monitoring comprises information that can influence patient management. Patient-controlled EEG home-monitoring is feasible as part of routine care for neurological outpatients as its technical efficacy and practical feasibility are shown and significantly positive effects on patient management are evidenced.


Mechanical properties and aesthetics of FRP orthodontic wire fabricated by hot drawing.
The FRP wires 0.5 mm in diameter with a multiple fiber structure were fabricated by drawing the fiber polymer complex at 250 degrees C for an esthetic, transparent orthodontic wire. Biocompatible CaO-P2O5-SiO2-Al2O3 (CPSA) glass fibers of 8-20 microm in diameter were oriented unidirectionally in the longitudinal direction in PMMA matrix. The mechanical properties were investigated by 3-point flexural test. The FRP wire showed sufficient strength and a very good elastic recovery after deformation. Young's modulus and the flexural load at deflection 1 mm were nearly independent of the fiber diameter and linearly increased with the fiber fraction. The dependence on fiber fraction obeys well the rule of mixture. This FRP wire could cover the range of strength corresponding to the conventional metal orthodontic wires from Ni-Ti used in the initial stage of orthodontic treatments to Co-Cr used in the final stage by changing the volume ratio of glass fibers with the same external diameter. The estheticity in external appearance was excellent. Thus the new FRP wire can satisfy both mechanical properties necessary for an orthodontic wire and enough estheticity, which was not possible for the conventional metal wire.


Cytomegalovirus infection of the ileoanal pouch: clinical characteristics and outcomes.
Up to 30% of cases of pouchitis are felt to have a secondary cause. Cytomegalovirus (CMV) may represent a possible etiopathological agent. Here, we report our experience with CMV involvement of the pouch, including risk factors, clinical features, and pouch outcomes in patients with inflammatory bowel disease after proctocolectomy with ileal pouch-anal anastomosis. The pathology database at Mayo Clinic in Rochester was searched between January 1995 and October 2012 for patients with a tissue diagnosis of CMV of the pouch following ileal pouch-anal anastomosis. Seven patients with CMV inclusions of the pouch were identified. The median age was 35 (range, 10-53) years, and the majority were female (71%). Five patients (71%) were on immunosuppressive medications including 4 who had undergone orthotopic liver transplantation for primary sclerosing cholangitis. The clinical presentation was similar among all patients: the majority had diarrhea (86%), fever (71%), and abdominal pain (57%). All had mucosal inflammation, with 71% having focal ulcerations in the pouch and 60% having inflammatory changes in the prepouch ileum. All patients improved with ganciclovir. None required pouch excision or had recurrent CMV infection. Three patients had recurrent nonspecific pouchitis. A high index of suspicion is needed to diagnose CMV of the pouch. An increase in stool frequency and fever in patients on immune suppression or in those who have failed empiric antibiotics should prompt assessment for CMV infection. Antiviral therapy seems to be effective, and postinfection pouch outcomes seem favorable, particularly in those presenting with their first episode of pouchitis.


Bevacizumab as an adjunct to vitreoretinal surgery for diabetic retinopathy in East Africa.
The purpose of this study is to evaluate the efficacy of preoperative intravitreal bevacizumab (IVB) for improving outcomes in vitrectomy for diabetic retinopathy-related non-clearing vitreous haemorrhage and/or tractional retinal detachment. Medical record from patients undergoing vitrectomy for proliferative diabetic retinopathy (PDR) were retrospectively analysed (2003-2011). From 2007, IVB (1.25 mg 2-4 days before operating) was used on all eyes. Eyes receiving IVB were compared with those that did not receive IVB. Intraoperative complications, reoperation rates, and final visual acuity were the core outcome measures. Data were analysed for 88 patients (101 eyes). In all, 41 (41%) patients had received IVB, whereas 60 (59%) patients had not. Significant intraoperative haemorrhage occurred in six eyes (10%) in the non-IVB group and in one (2.4%) IVB eyes (P=0.24). Silicon oil was used in 29 (48%) non-IVB eyes and in 11 (27%) IVB eyes (P=0.03). The non-IVB eyes underwent significantly more vitreoretinal reoperations (P=0.01) and were significantly more likely to lose two or more lines of vision at the final follow-up (P=0.03). The numbers needed to treat (NNT) blindness (<3/60) was four for non-IVB eyes and two for the IVB group. IVB reduces surgical complications, the use of silicon oil, and the need for further retinal surgery. The NNT to restore useful vision (≥3/60) to a blind eye were significantly lower in the IVB group. Vitreoretinal surgery for the complications of PDR is effective in an East African context, and IVB should be considered a valuable adjunct.


Expression of sodium-dependent phosphate (NadPi) transport in Xenopus laevis oocytes induced by mRNA from 1 alpha, 25-dihydroxyvitamin D3-treated rat osteoblast-like cells.
Injection of messenger ribonucleic acid (mRNA) isolated from 1 alpha, 25-dihydroxyvitamin D3-treated osteoblast-like (PyMS) cells leads to an enhanced sodium-dependent phosphate (NadPi) transport in Xenopus laevis oocytes, when compared to untreated cells. After mRNA size fractionation, mRNA with an average size of 2.2-3.8 kilobases showed up to a 1.8-fold stimulation of NadPi transport encoding either directly a NadPi transporter(s) or proteins controlling their activity. No hybridization was observed in Northern blots with RNA from rat bone or PyMS cells with the recently cloned rat renal brush border NadPi transporter NaPi-2; hybrid depletion with a NaPi-2 antisense oligonucleotide did not abolish the PyMS mRNA-induced NadPi transport in oocytes. We present the first evidence for functional expression in Xenopus laevis oocytes of a new type of NadPi transport system in bone cells, which is different from the renal type.


Two-year trends in cancer screening among low socioeconomic status women in an HMO-based high-deductible health plan.
Cancer screening is often fully covered under high-deductible health plans (HDHP), but low socioeconomic status (SES) women still might forego testing. To determine the impact of switching to a HDHP on breast and cervical cancer screening among women of low SES. Pre-post with comparison group. Four thousand one hundred and eighty-eight health plan members enrolled for one year before and up to two years after an employer-mandated switch from a traditional HMO to an HMO-based HDHP, compared with 9418 propensity score matched controls who remained in HMOs by employer choice. Both groups had low outpatient copayments. High-deductible members had full coverage of mammography and Pap smears, but $500 to $2000 individual deductibles for most other services. HMO members had full coverage of cancer screening and low copayments for other services without any deductible. We stratified analyses by SES. Transition to a HDHP. Annual breast and cervical cancer screening rates; rates of annual preventive outpatient visits. In follow-up years 1 and 2, low SES HDHP members experienced no statistically detectable changes in rates of breast cancer screening (ratio of change, 1.14, 95 % CI, [0.93,1.40] and 1.05, [0.80,1.37], respectively) or preventive visits (difference-in-differences, +1.9 %, [-11.9 %,+17.7 %] and +10.1 %, [-9.4 %,+33.7 %], respectively) relative to HMO counterparts. Similarly, among low SES HDHP members eligible for cervical cancer screening, no significant changes occurred in either screening rates (1.01, [0.86,1.20] and 1.08, [0.86,1.35]) or preventive visits (+0.2 %, [-11.4 %,+13.3 %] and -1.4 %, [-18.1,+18.6]). Patterns were statistically similar for high SES members. During two follow-up years, transition to an HMO-based HDHP with coverage of primary care visits and cancer screening did not lead to differentially lower rates of breast and cervical cancer screening or preventive visits for low SES women. Generalizability is limited to commercially insured women transitioning to HDHPs with low cost-sharing for cancer screening and primary care visits, a common design.


Multiple symmetric lipomatosis: treatment with liposuction.
Multiple symmetric lipomatosis is a fascinating disease of uncertain etiology that manifests as massive lipomatous deposits in specific areas of the body. The clinical syndrome and possible pathogenesis are discussed. Classic surgical lipectomy has been the only treatment thus far reported; we report a patient successfully treated with liposuction.


Mechanical effect of posterior wire or half-pin configuration on stabilization utilizing a model of circular external fixation of the foot.
Numerous studies have addressed biomechanical characteristics of circular external fixation of long bones. The objective of the present study was to evaluate stabilization of a simulated foot model using external fixation with either calcaneal tensioned stopper wires or half-pins. Fixation configurations of the calcaneus included two parallel wires, two wires crossing at either 30 degrees or 45 degrees, a 4-mm- and 5-mm-diameter single half-pin, or two half-pins inserted at a cross-angle of either 45 degrees or 90 degrees. All frames were tested in axial compression, anteroposterior (AP) bending, and mediolateral (ML) bending. An increase in wire cross-angle improved the axial and AP bending stabilization but had no influence on ML bending. Utilization of a single calcaneal half-pin instead of two cross-wires resulted in a considerable reduction in ML bending stabilization. Frame configurations with two half-pins substantially improved axial and ML bending stabilization. Due to the medial location of the metatarsal wire stopper, an increase in half-pin cross-angle significantly improved ML bending stiffness under lateral foot loading. Under the medial foot loading, however, the half-pin cross-angle had no effect on ML bending stabilization. Replacement of cross-wires with two half-pins significantly improved the AP bending stiffness only when the half-pin cross-angle was reduced to 45 degrees. In all modes of two half-pin frame loading, the half-pin diameter had a substantial effect on foot stabilization. Although the wire cross-angle, half-pin cross-angle, and half-pin diameter affect the stability of foot circular external fixation, the influence of these mechanical parameters on foot stabilization is dependent on the mode and location of loading. The results of the present mechanical testing can be utilized as a useful guideline for the optimization of circular external fixation of the foot.


Cryopreserved astrocytes maintain biological properties: Support of neuronal survival and differentiation.
Astrocytes, one of the main glial cell types, play critical roles in the central nervous system (CNS) development and function, including support of neuronal survival and differentiation, blood brain barrier formation, synapse homeostasis and injury response. Cell isolation and culture techniques have been proved to be a powerful tool to study astrocyte physiology and function. Due to financial constraints and rigid biosafety and ethics rules to use animal models, freezing techniques and the creation of cell banks emerged as alternatives to optimize the use of experimental animals. One of the main challenges, however, of these techniques is to guarantee that conserved cells keep their biological properties. In this work, we characterized morphologically and functionally murine secondary astrocyte cultures that have been submitted to freezing/thawing procedures. Morphological characterization of SAC (secondary astrocyte culture) and SFAC (secondary frozen-astrocyte culture) did not reveal significant differences on astrocyte morphology, confluence time and cell number along culture period. Functionally, SAC and SFAC did not reveal differences in their potential to support neuronal survival, maturation, neuritogenesis and synapse formation. Our results suggest that murine astrocytes that are submitted to freezing/thawing procedure maintain morphological and functional characteristics when compared with non-frozen astrocytes. Thus, this methodological approach is a valuable tool for in vitro research and might allow experimental optimization and reduction of animal use.


High dorsal column cordotomy plus subdiaphragmatic vagotomy prevents acute ionizing radiation sickness in cats.
Our purpose was to determine the effects on acute radiation sickness of interrupting afferent neural pathways that converge upon the medullary vomiting center but which bypass the emetic chemoreceptor trigger zone in the area postrema. A comparison was made of the vomiting response and other signs of sickness in three groups of chronic cats surgically prepared as follows: high spinal cord section of the dorsal columns, subdiaphragmatic vagotomy, and the combination of procedures. Every cat was exposed over the whole body to 45 Gy 60Co gamma-radiation which was effective in evoking emesis in 11 of 12 normal cats. Neither cordotomy alone (8 cats) nor vagotomy alone (2 cats) reliably blocked the vomiting response but they separately delayed its onset. On the other hand, the cordotomy prevented the loss of appetite and behavioral malaise that was invariably caused by the irradiation in normal cats. Finally, the combination of cordotomy and vagotomy protected all of 3 cats against the entire radiation syndrome. These cats then vomited appropriately in response to the injection of deslanoside which induces emesis through an action on the area postrema. Histological examination of the lower medulla revealed no damage of the area postrema resulting from the cordotomies. We conclude that acute radiation sickness in the cat is signaled through afferent neural pathways originating in the abdomen and that the area postrema does not participate in the causation of this syndrome.


Structural diversity of 1,2,4-diazaphospholide complexes with alkali metals.
Treatment of H[3,5-Ph2dp] (Hdp = 1H-1,2,4-diazaphosphole) with nBuLi or KH, or the reaction of K[3,5-tBu2dp] with an excess amount of O2, afforded the dimeric species [(eta2,eta1-3,5-Ph2dp)Li(THF)2]2 and the polymeric complexes [(eta2:eta4-3,5-Ph2dp)K(Et2O2]n, and [[(eta2:eta5-3,5-tBu2dp)K(THF)][eta2(N,N)-eta3(O,P,O)-3,5-tBu2dp-(O,O)O2K]]n, respectively.


Local protection for surgical implants.
In this issue of Chemistry & Biology, Jose and coworkers [1] describe the covalent attachment of vancomycin to a titanium surface. The tethered antibiotic is effective against S. aureus, suggesting that this material has the potential to complement clinical strategies for preventing infection following implant surgery.


Mechanism of gemini disulfide detergent mediated oxidative refolding of lysozyme in a new artificial chaperone system.
Gemini surfactants are a new class of surfactants that consist of two hydrophilic head groups and two hydrophobic tails separated by a spacer group. As the properties of geminis are different to their monomeric counterparts, a large number of applications have been investigated. Here we report on the use of a new class of gemini detergents containing a disulfide bond in the spacer (Det-SS-Det) for protein refolding. Using lysozyme as a model protein we could demonstrate that the disulfide gemini detergents allow oxidative refolding of the protein in the absence of any external redox system in an "artificial chaperone system". Refolding kinetics using gemini disulfide detergents differing in their hydrophobicity were analysed to determine the folding and aggregation rate constants. The results point to an important role of the transiently formed mixed disulfides between the protein and the detergent (Prot-SS-Det) in the oxidative refolding process of lysozyme.


Presence of immunoreactive atrial natriuretic peptide in nerve fibres and conduction cells in the conduction system of the bovine heart.
Previous findings of atrial (A-type) natriuretic peptide (ANP) in nervous tissue, such as the brain and the superior cervical ganglia, led us to investigate the possible occurrence of ANP in nervous tissue in the heart. The distribution of ANP in the bovine heart, particularly its conduction system, was examined by the use of immunohistochemical methods and an antiserum against alpha-hANP. ANP immunoreactivity was frequently detected in atrial myocytes and in the Purkinje fibres of the AV-bundle, and was sometimes seen in the Purkinje fibres of the bundle branches and their ramifications. On the other hand, ANP immunoreactivity was never seen in the conduction cells of SA- and AV-nodes. ANP immunoreactivity was also detected in small nerve-fibre varicosities, mainly in the AV-node and AV-bundle. Most of these varicosities were located in the proximity of the conduction cells, but some occurred close to fine blood vessels or in the walls of arterioles. These observations show for the first time that ANP immunoreactivity is present not only in atrial myocytes and conduction cells but also in nerve-fibre varicosities in the conduction system. The observations suggest that ANP may act as a neuromodulator and/or neurotransmitter in the conduction system.


Investigation of biocatalytic production of lactosucrose and fructooligosaccharides using levansucrases and dairy by-products as starting materials.
Selected levansucrases (LSs) were investigated for their ability to catalyze the transfructosylation of lactose/sucrose into lactosucrose and fructooligosaccharides (FOSs). Additionally, dairy by-products, including whey permeate (WP) and milk permeate (MP), were assessed for their effectiveness as lactose sources. LSs from Gluconobacter oxydans (LS1), Vibrio natriegens (LS2), Novosphingobium aromaticivorans (LS3), and Burkholderia graminis (LS4) were utilized in three transfructosylation reactions that combined sucrose with either lactose, WP, or MP. All LSs demonstrated a higher transfructosylation activity than hydrolytic one, except for V. natriegens LS2 in the presence of sucrose and MP/sucrose. Furthermore, the bioconversion efficiency of lactose/sucrose into lactosucrose and FOSs exhibited varying time courses and end-product profiles. Both the acceptor specificity of LS and the thermodynamic equilibrium of its reaction modulated the end-product profile. V. natriegens LS2 resulted in the highest lactosucrose production of 328 and 251 g/L with lactose/sucrose and WP/sucrose, respectively. Our results revealed the potential of LS-catalyzed transfructosylation for the biocatalytic production of both lactosucrose and FOSs from abundant biomasses.


Mitochondria-localized in situ generation of rhodamine photocage with fluorescence turn-on enabling cancer cell-specific drug delivery triggered by green light.
In this work, we have developed a rhodamine dye based water-soluble, mitochondria-indicating photocage, which gets activated in the mitochondria producing a fluorescent signal and on-demand releases the caged anticancer drug, chlorambucil, in the cancer cells selectively upon irradiation of green light.


Substitution of DNA-contacting amino acids with functional variants in the Gata-1 zinc finger: a structurally and phylogenetically guided mutagenesis.
DNA-binding functionality among transcription factor proteins is afforded by a number of structural motifs, such as the helix-turn-helix, helix-loop-helix, and zinc finger domains. The common thread among these diverse structures is their sequence-specific binding to essential promoter or other genetic regulatory sequences with high selectivity and affinity. One such motif, present in a wide range of organisms from bacteria to vertebrates, is the Gata-type zinc finger. This family of DNA-binding proteins is characterized by the presence of one or two (Cys)(4) metal binding sites which recognize the protein's eponymous binding site, GATA. Unlike other conserved DNA-binding domains, Gata proteins appear to be restricted to binding consensus GATA sequences, or near variations, in DNA. Since the architecture of the Gata finger seems built around recognizing this particular sequence, we set out to define the allowable range of amino acid substitutions along the DNA-binding surface of a Gata finger that could continue to support sequence-specific DNA-binding activity. Accordingly, we set up a one-hybrid screen in yeast based on the chicken Gata-1 C-terminal zinc finger. Mutant libraries were generated at five amino acids identified in the Gata-DNA structure as likely to mediate sequence-specific contacts between the Gata finger and DNA. These libraries were designed to give as exhaustive amino acid coverage as possible such that almost all alternative amino acids were screened at each of the five probed positions. Screening and characterization of these libraries revealed several functional amino acid substitutions at two leucines which contact the DNA at the 3' and 5' flanks of the GATA binding site, but no functional substituents for amino acids near the core of the binding site. This pattern is consistent with amino acid sequences of known DNA-binding Gata fingers.


Familial basal cell nevus syndrome.
The basal cell nevus syndrome is characterized by multiple basal cell nevi and basal cell carcinoma, cysts of the jaw, anomalies of ribs and spine, abnormal calcifications, and additional anomalies of the facial skull. A German family is described with manifestations of the syndrome in the mother and her three daughters. Expressivity was variable, in part due to age effects. The observation conforms to the assumed autosomal dominant mode of inheritance with high penetrance.


Assessment of carotid artery dose in the treatment of nasopharyngeal cancer with IMRT versus conventional radiotherapy.
To determine the radiation dose to the carotid artery in nasopharyngeal cancer patients treated with intensity-modulated radiotherapy (IMRT) and to compare it to the dose delivered by a conventional three-field (3F) technique. Sixteen patients with nasopharyngeal cancer who were treated at UCSF with IMRT were selected for this analysis. 3F plans were reconstructed for comparison. The carotid arteries were retrospectively contoured, and the dose received by each of the 32 carotid arteries was determined for both IMRT and 3F plans. A subset of 8 patients with N0/N1 nodal disease was selected for IMRT replanning using additional constraints to reduce the dose to the arteries. Using the standard prescription doses for IMRT and 3F plans, the dose delivered to 95% of the tumor volume was significantly higher in the IMRT plans, reflecting the greater conformality of this technique. The median mean dose to the carotid arteries was 65.7Gy with IMRT vs. 58.4Gy with 3F (p<0.001). After the application of dose constraints to the carotid arteries, it was possible to reduce the mean carotid dose to 54Gy in the IMRT replans. IMRT achieves a higher tumoricidal dose and superior clinical target volume coverage, but results in an increase in the carotid artery dose as compared to conventional 3F technique. With careful IMRT planning, it is possible to constrain the carotid dose for a subset of patients with low-risk neck disease. Further study is necessary to quantify the long-term clinical impact of this intervention.