Differential induction of glutathione transferase isoenzymes of mice stomach by diallyl sulfide, a naturally occurring anticarcinogen.
Diallyl sulfide (DAS), an organosulfur compound identified as the flavor component in garlic, has been shown to inhibit chemically induced neoplasia of forestomach and lung in mice. Even though the exact mechanism(s) of anti-neoplastic activity of DAS is not known, several independent studies suggest that this effect may, at least in part, be due to the elevation of glutathione-S-transferase (GST) activity. To gain further insight into the mechanism(s) of anti-carcinogenic activity of DAS, we have determined effect of orally administered DAS (25, 50 and 75 mumol) on levels of alpha, mu and pi class GSTs and glutathione (GSH) peroxidase and GSH reductase activities of female A/J mice stomach. Western blotting revealed presence of alpha, mu and pi class GSTs in mice stomach. A significant increase in all the three classes of GSTs was observed in the stomach of mice treated with DAS. Maximum increase in GST alpha and pi was evident by treating the animals with 75 mumol DAS whereas maximum induction of GST mu occurred after treating mice with 50 mumol DAS. GSH peroxidase activity towards t-butyl-hydroperoxide increased in a dose-dependent fashion in the mice stomach treated with DAS. Even though this activity towards hydrogen peroxide was similar in mice treated with 50 or 75 mumol DAS, these values were significantly higher than that of the control. GSH reductase was also elevated in the stomach of mice treated with 75 mumol DAS. These results suggest that DAS may exert anti-neoplastic effect by modulating GSH dependent detoxification enzymes.


Effectiveness of photobiomodulation in the treatment of primary burning mouth syndrome-a systematic review and meta-analysis.
To evaluate the effectiveness of photobiomodulation (PBM) on primary burning mouth syndrome (pBMS). We searched Chinese and English studies published before February 10, 2020. The databases used include PubMed, EMBASE, the Cochrane Library, Web of Science, Wanfang Database, and China National Knowledge Infrastructure (CNKI). Randomized controlled clinical trials (RCTs) that used the PBM to treat pBMS and reported specific treatment outcomes were considered for inclusion. We eventually included 12 RCTs, and 574 samples were included in these studies. The primary outcomes investigated were pain reduction and life quality improvement. A meta-analysis performed on 9 groups in 5 trials showed that PBM was effective in reducing pain compared with placebo (MD - 1.86, 95% CI - 2.59 to - 1.13, Z = 4.99, P < 0.00001). Meta-analysis was also performed on 7 groups in 4 trials and showed that PBM was effective in improving life quality compared with placebo (MD - 3.43, 95% CI - 5.11 to - 1.75, Z = 4.00, P < 0.0001). Qualitative analysis of the included RCTs found that PBM might also play a role in the decrease of TNF-α and IL-6 in saliva. Three studies that compared PBM with medications were evaluated by descriptive analysis. None of the treatment-related adverse event was reported. Up to date, PBM appears to have an effect on pain reduction and life quality improvement in pBMS patients. However, more evidence is still required to warrant its efficacy and safety in treating pBMS.


Molecular cloning and chromatin structure analysis of the murine alpha1(I) collagen gene domain.
We have isolated molecular clones of genomic mouse DNA spanning 55 kb, including the entire coding region of the murine alpha1(I) collagen (Col1a1) gene and 24 kb of 5' and 13 kb of 3'-flanking sequences, and have performed a detailed chromatin structure analysis of these sequences. Several new DNase-I-hypersensitive sites were identified. The distal 5'-flanking region contains two clusters of DNase-I-hypersensitive sites located between 7 and 8 kb and between 15 and 20 kb upstream of the start site of transcription, respectively. Several of these sites were shown to be present in collagen-producing, but not in non-producing cells, indicating that they are associated with transcription of the gene and may function in its regulation. One strong constitutive DNase-I-hypersensitive site at -18.5 kb was also cleaved by endogenous nucleases. The 3'-flanking region of the gene contains a DNase-I-hypersensitive site located 6 kb downstream of the end of the gene, as well as sequences that can induce a non-B DNA structure. Because these latter sequences coincide with DNase-I-hypersensitive sites in the homologous human gene, our results suggest that some regulatory elements may play a role in gene regulation, not by specific protein-DNA interactions but by virtue of their ability to induce a non-B DNA structure and/or an alternate chromatin conformation. A comparison of the murine and human Col1a1 domains shows a similar, although not identical, distribution of DNase-I-hypersensitive sites, indicating a conserved arrangement of regulatory elements. Our results strongly suggest that these new sites constitute regulatory elements which are involved in the transcriptional regulation and/or chromatin loop organization of the Col1a1 gene, and they are now amenable for functional analyses.


Effect of ultrasound-assisted alkaline treatment on functional property modifications of faba bean protein.
This study investigated alkaline shifting, ultrasonication, and their combination on functional modifications of faba bean protein isolate (FPI). The protein characterization revealed that the combined alkaline shifting/ultrasound treatment dissociated large aggregates of FPI into small ones, resulting in increased protein surface hydrophobicity and decreased free sulfhydryl groups. Such structural changes significantly increased FPI solubility from 12.2 to 40.4% to more than 95% at pH 3 and 7 and more than 80% at pH 6. Furthermore, the FPI modified by the combined alkaline shifting/ultrasound treatment produced emulsions with smaller particle size and superior stability at pH values 3 and 7. Likewise, a striking enhancement in foaming capacity (from 93% to 306-386%) and stability (from 10 s to 473-974 s) was achieved by the combined treatment. This study suggested that the ultrasound-assisted alkaline shifting treatment could be considered an effective method for improving FPI functional properties.


Recommendations of the AFU Infectious Diseases Committee on the prevention, diagnosis and treatment of infections of endo-ureteral equipment.
No recent national guidelines exist regarding the management of urinary tract infections (UTIs) in the presence of ureteral stent. This situation could lead to discrepancies in clinical management and less opportunity for a favorable patient's outcome. All available data published on Medline® between 1998 and 2018 were systematically searched and reviewed. All papers assessing adult patients carrying ureteral stent were included for analysis. After studies critical analysis, national guidelines for clinical management were elaborated in order to answer clinical questions. A total of 451 articles were identified, of which 58 have been included. The prevalence of urinary tract infections in the presence of ureteral stent remains unknown. After 3 months, all endo-ureteral devices were colonized on microbiological study. These patients also presented a positive urine culture in 25 to 70% of the cases, often polymicrobial. Staphylococci, E. coli, Klebsiella, Pseudomonas, Enterococcus and Candida were the commonest micro-organisms responsible for urinary colonization or infection. The risk of UTI on endo-ureteral devices seemed higher the longer it stayed implanted. There is no justification in the literature to recommend a systematic change of endo-ureteral devices following a urinary tract infection. The existing literature is rich but of poor methodological quality, and therefore does not allow to draw robust conclusions. The greatest difficulty faced in this work was to accurately differentiate urinary colonizations from true infections, including clinical symptoms and not only microbiological results. These guidelines propose a standardized management of such common clinical situations. Well-designed studies are needed to upgrade the level of evidence of these guidelines.


Histamine metabolism. II. Cellular and subcellular localization of the catabolic enzymes, histaminase and histamine methyl transferase, in human leukocytes.
Histaminase (EC 188.8.131.52) activity has been demonstrated in human eosinophils and neutrophils, but not in mononuclear cells, with the use of a new and specific thin-layer radiochromatographic enzyme assay. Leukocyte histaminase was physicochemically and functionally similar to histaminase isolated from human placenta and was principally localized to the 27,000-g granule-rich fraction of eosinophil and neutrophil homogenates. Histamine methyl transferase (EC 18.104.22.168), on the other hand, was detected in monocytes but not in granulocytes, eosinophils, lymphocytes, or platelets, and was localized solely to the 100,000-g cell sap supernatant fraction. These data suggest a role of human leukocytes in the catabolism of histamine and therefore in the modulation of histamine-mediated inflammatory reactions.


Novel Approaches Used in Ovarian Tissue Transplantation for Fertility Preservation: Focus on Tissue Engineering Approaches and Angiogenesis Capacity.
Due to the impact of the modern lifestyle, female infertility has been reduced because of different reasons. For example, in combined chemotherapeutic therapies, a small fraction of cancer survivors has faced different post-complications and side effects such as infertility. Besides, in modern society, delayed age of childbearing has also affected fertility. Nowadays, ovarian tissue cryopreservation and transplantation (OTC/T) is considered one of the appropriate strategies for the restoration of ovarian tissue and bioactivity in patients with the loss of reproductive function. In this regard, several procedures have been considered to improve the efficacy and safety of OTT. Among them, a surgical approach is used to transplant ovaries into the optimal sites, but the existence of ischemic changes and lack of appropriate revascularization can lead to bulk follicular atresia. Besides, the role of OTC/T is limited in women of advanced maternal age undergoing lifesaving chemo-radiation. As a correlate, the development of de novo approaches with efficacious regenerative outcomes is highly welcomed. Tissue engineering shows high therapeutic potentialities to restore fertility in males and females using the combination of biomaterials, cells, and growth factors. Unfortunately, most synthetic and natural materials are at the experimental stage and only the efficacy has been properly evaluated in limited cases. Along with these descriptions, strategies associated with the induction of angiogenesis in transplanted ovaries can diminish the injuries associated with ischemic changes. In this review, the authors tried to summarize recent techniques, especially tissue engineering approaches for improving ovarian function and fertility by focusing on angiogenesis and neovascularization.


Does a Crossover Age Effect Exist for African American and Hispanic Binge Drinkers? Findings from the 2010 to 2013 National Study on Drug Use and Health.
Among general population studies, lower rates of binge drinking tend to be found among African Americans and Hispanics compared to Whites. However, among older adult populations, minority groups have been shown to be at higher risk for binge drinking, suggesting the presence of a crossover effect from low to high risk as a function of age. To date, limited research has examined the crossover effect among African American and Hispanic populations compared to non-Hispanic Whites across large developmental time frames or explored variation in risk based on income or gender. This study aimed to fill these gaps in the literature. Data were compiled from the 2010 to 2013 National Survey on Drug Use and Health surveys, which provide annual, nationally representative data on substance use behaviors among individuals aged 12 and older. Hispanic, non-Hispanic African American, and non-Hispanic White respondents were included (N = 205,198) in the analyses. A crossover effect was found for African American males and females among the lowest income level (i.e., incomes less than $20,000). Specifically, after controlling for education and marital status, compared to Whites, risk for binge drinking was lower for African American males at ages 18 to 24 and for females at ages 18 to 34, but higher for both African American males and females at ages 50 to 64. No crossover effect was found for Hispanic respondents. Although African Americans are generally at lower risk for binge drinking, risk appears to increase disproportionately with age among those who are impoverished. Explanatory factors, such as social determinants of health prevalent within low-income African American communities (e.g., lower education, violence exposure, housing insecurity) and potential areas for intervention programming are discussed.


The significance of fetal electrocardiography in the diagnosis of intrauterine bradyarrhythmia.
From 11 cases of fetal bradycardia diagnosed by monitoring of 130 fetal electrocardiograms (ECG's) in high-risk pregnancies we have presented our experience in three selected cases of fetal bradyarrthythmia. Case 1 revealed on ECG blocked atrial premature beats simulating an extreme sinus bradycardia sequentially followed by conducted atrial premature beats. In case 2 we diagnosed ventricular premature beats in the form of persistent bigeminy which was controlled by intravenous propranolol. The last case illustrated the phenomenon of aberrant ventricular conduction known to occur in adult cardiology. The electrophysiologic basis of the variable arrhythmias was discussed. Detailed analysis of repeated direct fetal ECG's provided us with the diagnosis and understanding of the electrophysiologic mechanisms underlying the rhythm disturbances. This consequently determined the pharmacologic therapy and the obstetric approach relevant to each case. We have shown that by direct fetal electrocardiography it is possible to analyze accurately the rhythm disturbances. Persistent fetal bradycardia does not always signify fetal distress. We hope that this will lead to closer teamwork between the obstetrician and the cardiologist which will give an impetus to the future development of "fetal cardiology," thereby enhancing our understanding of the electrophysiology of the fetal heart.


Assembly of a pangenome for global cattle reveals missing sequences and novel structural variations, providing new insights into their diversity and evolutionary history.
A cattle pangenome representation was created based on the genome sequences of 898 cattle representing 57 breeds. The pangenome identified 83 Mb of sequence not found in the cattle reference genome, representing 3.1% novel sequence compared with the 2.71-Gb reference. A catalog of structural variants developed from this cattle population identified 3.3 million deletions, 0.12 million inversions, and 0.18 million duplications. Estimates of breed ancestry and hybridization between cattle breeds using insertion/deletions as markers were similar to those produced by single nucleotide polymorphism-based analysis. Hundreds of deletions were observed to have stratification based on subspecies and breed. For example, an insertion of a Bov-tA1 repeat element was identified in the first intron of the APPL2 gene and correlated with cattle breed geographic distribution. This insertion falls within a segment overlapping predicted enhancer and promoter regions of the gene, and could affect important traits such as immune response, olfactory functions, cell proliferation, and glucose metabolism in muscle. The results indicate that pangenomes are a valuable resource for studying diversity and evolutionary history, and help to delineate how domestication, trait-based breeding, and adaptive introgression have shaped the cattle genome.


p53 tumor-suppressor gene: clues to molecular carcinogenesis.
The tumor-suppressor gene product p53 is clearly a component in several biochemical pathways, including transcription, DNA repair, genomic stability, cell-cycle control and apoptosis, that are central to human carcinogenesis. The p53 is functionally inactivated by mutational, viral, and cellular mechanisms in the majority of human cancers. Analysis of the spectrum of p53 mutations provides clues to the etiology and molecular pathogenesis of cancer. Recent insight into the p53-mediated biochemical pathways of cell-cycle arrest and apoptosis has provided further understanding of the mechanisms related to p53-mediated tumor suppression. This insight in turn may provide the potential molecular targets for the development of rational multimodality cancer therapy, including chemo-, immuno-, and gene-therapeutic strategies. The convergence of previously parallel lines of basic, clinical, and epidemiologic investigation may provide an opportunity to transfer research findings rapidly from the laboratory to the clinic.


In vivo immunomodulation of IL6 signaling in a murine multiple trauma model.
A significant number of trauma patients die during the ICU phase of care because of a severe immune response. Interleukin-6 (IL6) plays a central role within that immune response, signaling through a membrane-bound (IL6-R) and a soluble IL6 receptor (sIL6-R). IL6 and the sIL6-R can form an agonistic IL6/sIL6-R-complex, activating numerous cells that are usually not IL6 responsive, a process called trans-signaling. We attempted to demonstrate that modulation of the IL6 signaling (classic signaling and trans-signaling) can attenuate the devastating immune response after trauma in a murine multiple trauma model. Mice were allocated to three study arms: sham, fracture or polytrauma. Half of the animals had the application of an IL6-R antibody following an intervention. After a pre-set time, blood samples were analysed for IL6 and sIL6-R serum levels, organs were analysed for neutrophil infiltration and end organ damage was evaluated. IL6 and sIL6-R showed a rapid peak after fracture, and much more markedly after polytrauma. These parameters were reduced significantly by globally blocking IL6 signaling via IL6-R antibody (Mab) application. Shock organ analysis also illustrated significant neutrophil infiltration following polytrauma, which was also abated via IL6-R Mab application. Furthermore, end organ damage was reduced by IL6-R Mab application. The study results prove the regulatory role of IL6 signaling pathways in polytrauma, with haemorrhagic shock being a major trigger of inflammatory response. Modulation of IL6 signaling shows promise in the prevention of adverse events like organ failure following major trauma and might be a target for in vivo immunomodulation to reduce mortality in severely injured patients, but further evaluation regarding classic IL6 signaling and IL6 trans-signaling is needed.


Protective immunity against spring viremia of carp virus by mannose modified chitosan loaded DNA vaccine.
Spring viremia of carp virus (SVCV) usually be considered as one of the serious in viral diseases of aquaculture, and DNA vaccine with novel delivery mechanism or adjuvant has proven to be a promising and effective strategy to control aquatic animal diseases. In this study, the mannose-modified chitosan, a carrier system for vaccine delivery, were used to developed a chitosan-encapsulated DNA vaccine (CS-M-G) against SVCV, then investigated immune response induced by the vaccine. Our results showed that CS-M-G was confirmed the spherical or elliptical with even distribution and ranging from approximately 50 to 150 nm in size, the expression of the antigen gene could still be detected after 21 d post vaccination. The CS-M-G induces the highest antibody levels in the 20 μg dose group which is about 3 times than naked plasmid group at 21 d post vaccination, and still hold a higher level than control group at 28 d post vaccination. On the side, strongest protection with relative percent survival of 62.1% in the 20 μg CS-M-G group, which could produce significantly higher enzyme activities and up-regulated expression of immune-associated genes than control group. Thus, our results indicate that DNA vaccine loaded with mannose-modified chitosan induces strong immune response and provided an effective protection against SVCV infection, may be helpful and extended for developing more aquatic animal vaccines in the future.


Chronic systolic heart failure, guideline-directed medical therapy, and systemic hypotension-less pressure but maybe more risk (does this clinical scenario need more discussion?).
Many clinical trials have demonstrated the survival benefit of medication regimens that modulate the neurohormonal activation that occurs with chronic heart failure (HF). These medications, however, also commonly lower systemic blood pressure (BP). Low arterial BP in patients with chronic HF has been shown to be an independent predictor of increased mortality. Given this apparent paradox in therapeutic goals (treat aggressively but keep BP from going too low), how low should we allow systemic BP to go as a result of our medication regimens before we compromise the proven benefits of such drug therapy? Or is the association between the BP-lowering effects of standard therapy and outcomes in HF even meaningful clinically? It is from this perspective that the merits, potential clinical implications, and the relevant published literature pertaining to this patient and practice management issue will be discussed.


The transcriptome analysis on urea response mechanism in the process of ergosterol synthesis by Cordyceps cicadae.
Nitrogen source is required for the growth of Cordyceps cicadae and involved in the regulation of metabolite synthesis. In order to further investigate the regulatory effects of nitrogen sources on the ergosterol synthesis by C. cicadae. We first confirmed that urea could significantly increase the ergosterol synthesis. The transcriptome analysis showed that compared with biomass cultured in the control fermentation medium (CFM), 1340 differentially expressed genes (DEGs) were obtained by Gene Ontology (GO) annotation, and 312 DEGs were obtained by Kyoto Encyclopedia of Genes and Genomes (KEGG) annotation from the biomass cultured in CFM + CO(NH2)2. Urea up-regulated D-3-phosphoglycerate dehydrogenase gene transcription level and down-regulated enolase and L-serine/L-threonine ammonialyase gene transcription level, increased serine synthesis, allosterically activate pyruvate kinase, to promote the synthesis of pyruvate and CH3CO ~ SCOA, the primer of ergosterol; Urea increase the genes transcription related with ergosterol synthesis by up-regulating the steroid regulatory element binding protein gene transcription levels. The transcriptome results were provided by those of qRT-PCR. Collectively, our finding provided valuable insights into the regulatory effect of nitrogen source on the ergosterol synthesis by C. cicadae.


A microassay method for neurotoxic esterase determinations.
A microtiter plate reader with an associated computer to average triplicate samples and subtract blanks was used for reading and calculating neurotoxic esterase (NTE, also known as neuropathy target esterase) activities in spinal cord regions of hens 4 hr after administration of diisopropylphosphorofluoridate (DFP, 0.5 mg/kg sc). Although NTE inhibition is an early indicator of organophosphorus ester-induced delayed neuropathy. DFP-induced inhibition was not greater in regions of the spinal cord where pathological changes are most notable. Acetylcholinesterase (AChE) activities and protein determinations were also done on these tissues using microassay methods. DFP-induced AChE inhibition was similar to NTE inhibition. In addition to the capability to be used for small regional esterase activity measurements, the microassay was advantageous because the number of samples incorporated into a single assay was increased and the time needed for the NTE assay was reduced by 50%. Total volume of incubate in each well was 0.3 ml; the incubate contained 1/20 quantities of sample and reagents necessary in more conventional assays. Validation of the microassay was performed by comparison with more conventional assays when measuring inhibition of NTE and AChE in brains of control and experimental hens of two different genetic strains (B13B13 and B21B21 white leghorns). Experimental birds were given DFP, 0.5 mg/kg sc, 24 hr before samples were collected. NTE activities in brains of control hens were similar using both types of NTE analytical procedures. Percentage inhibition of NTE caused by DFP was within 4% using both assay procedures in both strains of hens.(ABSTRACT TRUNCATED AT 250 WORDS)


Quantification of rotavirus-like particles by gel permeation chromatography.
There is a lack of accurate and practical methods that require only small amounts of sample for quantifying virus-like particles (VLP). In this work, gel permeation (GP) HPLC was used to quantify double-layered rotavirus-like particles (dlRLP) produced in insect cells. The proposed methodology utilized two columns in series (pore sizes of 200 and 50 nm) and had a high precision (relative standard deviation below 5%). GP-HPLC not only allowed the routine quantification of dlRLP, but also of assembly intermediaries and other viral structures present in the samples. For the first time, kinetics of dlRLP accumulation could be followed. This methodology is valuable for designing new production processes and for optimizing dlRLP monitoring.


Regional cell density distribution and oxygen consumption rates in porcine TMJ discs: an explant study.
To determine the regional cell density distribution and basal oxygen consumption rates (based on tissue volume and cell number) of temporomandibular joint (TMJ) discs and further examine the impact of oxygen tension on these rates. TMJ discs from pigs aged 6-8 months were divided into five regions: anterior, intermediate, posterior, lateral and medial. The cell density was determined using confocal laser scanning microscopy. The change in oxygen tension was recorded while TMJ disc explants were cultured in sealed metabolism chambers. The volume based oxygen consumption rate of explants was determined by theoretical curve-fitting of the recorded oxygen tension data with the Michaelis-Menten equation. The rate on a per-cell basis was calculated based on the cell density measurements and volume based rate measured in another group of discs. The overall cell density [mean, 95% confidence interval (CI)] was 51.3 (21.3-81.3) × 10(6) cells/mL wet tissue. Along the anteroposterior axis, the anterior band had 25.5% higher cell density than the intermediate zone (P<0.02) and 29.1% higher than the posterior band (P<0.008). Along the mediolateral axes, the medial region had 26.2% higher cell density than the intermediate zone (P<0.04) and 25.4% higher than the lateral region (P<0.045). The overall volume and cell based maximum oxygen consumption rates were 1.44 (0.44-2.44) μmol/mL wet tissue/h and 28.7 (12.2-45.2)nmol/10(6)cells/h, respectively. The central regions (intermediate, lateral, and medial) had significantly higher volume based (P<0.02) and cell based (P<0.005) oxygen consumption rates than the anterior and posterior bands. At high oxygen tension, the oxygen consumption rate remained constant, but dropped as oxygen tension fell below 5%. The TMJ disc had higher cell density and oxygen consumption rates than articular cartilage reported in the literature. These results suggest that a steeper oxygen gradient may exist in the TMJ disc and may be vulnerable to pathological events that impede nutrient supply.


Reduction in the level of immobilization in forced swim test and ethanol intake in rats by oxygen therapy.
Experiments replicated the previous finding that rats with high immobilization time in the forced swim test (passive rats) consumed more 15% ethanol solution in a free choice situation with tap water than rats with active behavior (active rats). Exposure of passive rats to oxygen under normal and elevated (2 ata) pressure resulted in the decrease in immobilization scores in the forced swim test as well as reduction in alcohol consumption and preference.


Regulation of electron transport is essential for photosystem I stability and plant growth.
Photosynthetic electron transport is regulated by cyclic and pseudocyclic electron flow (CEF and PCEF) to maintain the balance between light availability and metabolic demands. CEF transfers electrons from photosystem I to the plastoquinone pool with two mechanisms, dependent either on PGR5/PGRL1 or on the type I NADH dehydrogenase-like (NDH) complex. PCEF uses electrons from photosystem I to reduce oxygen and in many groups of photosynthetic organisms, but remarkably not in angiosperms, it is catalyzed by flavodiiron proteins (FLVs). In this study, Physcomitrella patens plants depleted in PGRL1, NDH and FLVs in different combinations were generated and characterized, showing that all these mechanisms are active in this moss. Surprisingly, in contrast to flowering plants, Physcomitrella patens can cope with the simultaneous inactivation of PGR5- and NDH-dependent CEF but, when FLVs are also depleted, plants show strong growth reduction and photosynthetic activity is drastically reduced. The results demonstrate that mechanisms for modulation of photosynthetic electron transport have large functional overlap but are together indispensable to protect photosystem I from damage and they are an essential component for photosynthesis in any light regime.


Nanoscale cobalt-based metal-organic framework impairs learning and memory ability without noticeable general toxicity: First in vivo evidence.
Metal-organic frameworks (MOFs) exhibit broad potential applications in the environmental, biomedical, catalyst, and energy fields. However, the currently existing data hardly shed light on their health risks before the MOFs' large-scale usage. In this context, we exploratively investigated the in vivo fate and effect of one representative cobalt-based zeolitic imidazolate framework (ZIF-67) at the nano- (60 nm) and submicron- (890 nm) scales. Different from submicron-scale ZIF-67 showing better biosafety, nanoscale particles manifested a neurodegenerative risk at the dose of no general toxicity, evidenced by the impairment of learning and memory ability and disordered function of the neuropeptide signaling pathway in a rat model. The involvement of oxidative damage and inflammatory processes in the neurotoxicity induced by ZIF-67 was discussed as well. These findings not only provide a wake-up call for the prudent applications of MOFs but also provide insight into the better design and safer use of MOFs for broader applications.


Processing of bacteriophage T4 tRNAs: a precursor of species 1 RNA.
A precursor molecule of species 1 RNA, p2Sp1, that accumulates when an rne (RNase E-) mutant is infected with a T4 deletion mutant (delta 27) is also found after infection of an rne host mutant by different deletion mutants or wild type bacteriophage T4. Low levels of this molecule were also found in a wild-type host infected with a wild-type T4. This precursor molecule accumulates at higher concentrations at 43 degrees C as compared to 30 degrees C or 37 degrees C. Structural analysis of the precursor molecules from the different sources has shown a complete identity of p2Sp1 RNA isolated from the different sources. Therefore, we suggest that this precursor is a normal intermediate in processing of T4 tRNAs, and that it is unrelated to a particular T4 deletion strain. Since RNase E does not process this precursor, its accumulation in an rne mutant reflects an interaction between RNase E and the enzyme that processes this intermediate.


Distinguishing between isomeric neoxanthin and violaxanthin esters in yellow flower petals using liquid chromatography/photodiode array atmospheric pressure chemical ionization mass spectrometry and tandem mass spectrometry.
Liquid chromatography/photodiode array atmospheric pressure chemical ionization mass spectrometry (LC/PDA-APCI-MS) is used for the analysis of various carotenoid pigments in plants. Among them, it is difficult to distinguish between the isomeric violaxanthin/neoxanthin esters. The yellow pigments of tomato petals were extracted with acetone, and the extracts were kept at -30°C to allow the contaminating triacylglycerols to settle out physically. The supernatants were analyzed using LC/PDA-APCI-MS with a high-resolution orbitrap mass spectrometer for their exact masses. The expected carotenoid esters were calculated with the combination of carotenoids and fatty acids, and they were matched with the experimental exact masses. The fatty acid structures in the carotenoid esters were also identified using collision-induced dissociation (CID) tandem mass spectrometry (MS/MS). The isomeric violaxanthin/neoxanthin esters were distinguished using CID MS/MS from their in-source dehydrated product ions as pseudoprecursor ions. The in-source dehydrated ions [M - H2 O + H]+ of neoxanthin diesters predominated over their protonated molecules [M + H]+ in LC/MS. By contrast, the protonated molecules of violaxanthin diesters predominated. The 92 u loss product ions [M - H2 O - C7 H8 + H]+ were observed from the dehydrated violaxanthin diesters, but they were not generated from the dehydrated neoxanthin diesters in the CID MS/MS of their dehydrated pseudoprecursor ion [M - H2 O + H]+ . The allene allyl carbocation in neoxanthin diesters was generated from dehydration after preferential protonation at the hydroxy group. The epoxide group of violaxanthin diesters opens easily after protonation; however, the dehydration did not proceed at this stage. The 92 u loss of C7 H8 was explained by an intramolecular [2 + 2] cycloaddition, which proceeded preferentially in dehydrated violaxanthin diesters because the carbocations in the dehydrated species were conjugated to the polyene and those double bonds were depolarized during CID MS/MS. Therefore, the isomeric neoxanthin/violaxanthin diesters were distinguished using LC/PDA-APCI-MS and MS/MS. This method was a practical and useful method of profiling the carotenoid esters of the yellow petals.


The modulatory role of dopamine receptors in brain neuroinflammation.
Neuroinflammation is a general pathological feature of central nervous system (CNS) diseases, primarily caused by activation of astrocytes and microglia, as well as the infiltration of peripheral immune cells. Inhibition of neuroinflammation is an important strategy in the treatment of brain disorders. Dopamine (DA) receptor, a significant G protein-coupled receptor (GPCR), is classified into two families: D1-like (D1 and D5) and D2-like (D2, D3 and D4) receptor families, according to their downstream signaling pathways. Traditionally, DA receptor forms a wide variety of psychological activities and motor functions, such as voluntary movement, working memory and learning. Recently, the role of DA receptor in neuroinflammation has been investigated widely, mainly focusing on nucleotide-binding oligomerization domain-like receptor pyrin domain-containing 3 (NLRP3) inflammasome, renin-angiotensin system, αB-crystallin, as well as invading peripheral immune cells, including T cells, dendritic cells, macrophages and monocytes. This review briefly outlined the functions and signaling pathways of DA receptor subtypes as well as its role in inflammation-related glial cells, and subsequently summarized the mechanisms of DA receptors affecting neuroinflammation. Meaningfully, this article provided a theoretical basis for drug development targeting DA receptors in inflammation-related brain diseases.


Chiral capillary electrophoresis-mass spectrometry of tetrahydroisoquinoline-derived neurotoxins: observation of complex stereoisomerism.
Previous studies have shown that certain 1,2,3,4-tetrahydroisoquinoline derivatives (TIQs) are neurotoxins inducing Parkinsonism. Further, individual enantiomers of these toxins such as (R/S)-N-methylsalsolinol ((R/S)-NMSal) possess distinct neurotoxicological properties. In this work, a chiral capillary electrophoresis (CE) method with electrospray ionization-tandem mass spectrometric (ESI-MS/MS) detection was developed for the quantification of TIQ enantiomers. Enantioseparation was achieved with sulfated β-cyclodextrin (sulfated β-CD) as chiral selector. To avoid any potential contamination of MS ionization source by the non-volatile chiral selector, partial filling technique was deployed in the CE separation. TIQ derivatives, including (R/S)-6,7-dihydroxy-1-methy-TIQ (salsolinol, Sal), (R/S)-1-benzyl-TIQ (BTIQ), and (R/S)-NMSal, were base-line resolved with resolution values (R) ranging from 3 (for Sal) to 4.5 (for BTIQ), which were much better than those reported previously by HPLC methods. ESI-MS/MS detection of the resolved TIQ enantiomers was specific and sensitive (LOD=1.2 μM for Sal enantiomers). The proposed chiral CE-MS/MS method was used to study in vitro formation of (R/S)-NMSal. It was found that NMSal was formed from the incubation of epinine (a dopamine metabolite) with acetaldehyde (a metabolite of alcohol). More interestingly, four isomers of NMSal were separated and detected in the incubation solution. They were identified as (R)-e.e-NMSal, (R)-e.a-NMSal, (S)-e.e-NMSal, and (S)-e.a-NMSal. This was the first lab evidence that this Parkinsonian neurotoxin exists in multiple isomeric forms.


Medullary thyroid cancer outcomes in patients with undetectable versus normalized postoperative calcitonin levels.
Calcitonin (Ct) is a sensitive diagnostic biomarker and one of the most important prognostic factors in medullary thyroid cancer (MTC). This study aimed to evaluate progression-free survival and recurrence rates of MTC associated with undetectable compared with normalized serum Ct levels after surgery. This retrospective observational study included patients operated for MTC at the Digestive and Endocrine Surgery Department of Lyon Sud Hospital Centre between 2000 and 2019. Clinical and pathological factors were correlated with postoperative Ct concentrations. Undetectable and normalized Ct concentrations were defined as below 2 pg/ml and 2-10 pg/ml respectively. Overall, 176 patients were treated for MTC, and 127 were considered biochemically cured after surgery. Of these, 24 and 103 had normalized and undetectable Ct concentrations respectively. Patients with Ct level normalization had a 25 per cent risk of disease recurrence, compared with 3 per cent in patients with undetectable Ct levels after surgery. The presence of metastasis in two or more compartments was predictive of failure to achieve undetectable Ct concentrations after surgery and an increased risk of recurrence. Among patients with biochemically cured MTC, those with undetectable or normalized Ct concentrations after surgery had different risks of recurrence. Simply assessing postoperative Ct normalization can be falsely reassuring, and long-term follow-up is needed. Calcitonin (Ct) is a sensitive diagnostic biomarker and one of the most important prognostic factors for medullary thyroid cancer outcomes; however, the significance of postoperative Ct levels remains controversial. This study evaluated the differences between normal and undetectable postoperative Ct levels in patients who had undergone surgical treatment for medullary thyroid cancer. Patients who experienced postoperative Ct level normalization had a higher risk of disease recurrence than those with undetectable Ct levels after surgery.


Indomethacin enhances the proliferation of mitogen-stimulated T lymphocytes of homosexual males with persistent generalized lymphadenopathy.
The possibility that cyclooxygenase products of arachidonic acid might contribute to the defective T lymphocyte function of homosexual men with the reactive lymph node syndrome was examined in vitro. T lymphocyte proliferation, assessed by the uptake of [3H]thymidine after the addition of phytohemagglutin, was 72,870 +/- 66,816 counts per minute (mean +/- SD) for eight patients and 119,589 +/- 64,913 counts per minute for 30 controls (P less than 0.05, Student's t test). Treatment with the cyclooxygenase inhibitor indomethacin increased the phytohemagglutin-induced proliferation of the T lymphocytes from five of eight patients, but none of 12 healthy homosexual and heterosexual control subjects. The production of prostaglandin E2 by T lymphocytes from six patients was 16.1 +/- 10.5 pg/5 X 10(6) cells/hr, as contrasted with that of 4.9 +/- 1.3 and 4.3 +/- 2.1 pg/5 X 10(6) cells for four healthy homosexual and six healthy heterosexual control subjects, respectively (P less than 0.01, Student's t test). The production of prostaglandin E2 by the patients' monocytes was normal. Abnormalities of the cyclooxygenase pathway of T lymphocytes of patients with the reactive lymph node syndrome may reflect an immunoregulatory defect, which predisposes to infections and may evolve into the more severe abnormalities of the acquired immune deficiency syndrome.


Tuning the demixing of colloid-polymer systems through the dispersing solvent.
We report measurements on fluid-fluid phase separation in a colloid-polymer mixture by means of confocal scanning laser microscopy and we show that we can access the various coarsening regimes by tuning the properties of the solvent. By increasing the viscosity of the solvent we are able to access the diffusive hydrodynamic regime of spinodal decomposition. By matching the density of the solvent and colloids we are able to grow structures to large length scales before they are destroyed by buoyancy forces. Tuning the solvent's density furthermore gives control over which phase flows up and down, illustrating the flexibility of this particular system.


Effectiveness of influenza vaccination during pregnancy to prevent severe infection in children under 6 months of age, Spain, 2017-2019.
Influenza vaccination is recommended to pregnant women in Spain to reduce the risk of influenza-related complications. Influenza related hospitalizations pose a significant disease burden in children every year. Although children below 6 months are too young to be vaccinated, they can receive protection against influenza through vaccination of their mothers during pregnancy. We estimated the effectiveness of maternal influenza vaccination to prevent influenza hospitalizations in infants under 6 months of age. This is a retrospective pilot study, using data from the Severe Hospitalized Confirmed Influenza Cases (SHCIC) surveillance system in seasons 2017/18 and 2018/19 in Spain. Maternal vaccination status during pregnancy was collected for cases in children 6 months and younger hospitalized with confirmed influenza infection. Influenza vaccine effectiveness was estimated using the screening method, by comparing the proportion of children with vaccinated mothers during pregnancy (proportion of cases vaccinated, PCV), with the vaccination coverage among pregnant women in Spain (proportion of population vaccinated, PPV). For all the study period, the PCV was 17% and the PPV was 35%. Influenza vaccination in mothers during pregnancy prevented influenza confirmed hospitalizations in infants aged 6 months and younger with a 61% (95%CI: 27-79%) effectiveness. In line with evidence from other countries, influenza vaccination during pregnancy protects infants up to 6 months of age from influenza hospitalizations in Spain. These results support current recommendations of influenza vaccination in pregnant women, and more studies are needed in Spain to confirm the double protection of maternal vaccination in mothers and infants.


Morphologically specific vestibular hair cell degeneration in the jerker mouse mutant.
The vestibular organs in homozygote (je/je) and heterozygote (je/+) 1- to 12-month-old jerker mouse mutants were analyzed morphologically using light microscopy and transmission and scanning electron microscopy. The pathological changes were initially confined to the surface structure of predominantly type I hair cells. The latter cells showed disintegration of the cuticular plate, disarray of stereocilia and finally fusion of sensory hairs. The hair cells were then expelled into the endolymphatic space. A notable finding was that most expelled hair cells still showed a rather normal ultrastructure of their cytoplasm and nuclei. The morphological type of pathology was the same in je/je and je/+ mutants. However, in je/je animals the degeneration occurred within 1-3 months after birth, whereas in je/+ mutants a slowly progressive degeneration lasted for up to 1 year of age or even more. The degeneration was more extensive in the two maculae than in the three cristae ampullares.


Narrative Review of Glycemic Management in People With Diabetes on Peritoneal Dialysis.
There is an increasing number of people with diabetes on peritoneal dialysis (PD) worldwide. However, there is a lack of guidelines and clinical recommendations for managing glucose control in people with diabetes on PD. The aim of this review is to provide a summary of the relevant literature and highlight key clinical considerations with practical aspects in the management of diabetes in people undergoing PD. A formal systematic review was not conducted because of the lack of sufficient and suitable clinical studies. A literature search was performed using PubMed, MEDLINE, Central, Google Scholar and ClinicalTrials.gov., from 1980 through February 2022. The search was limited to publications in English. This narrative review and related guidance have been developed jointly by diabetologists and nephrologists, who reviewed all available current global evidence regarding the management of diabetes in people on PD.We focus on the importance of individualized care for people with diabetes on PD, the burden of hypoglycemia, glycemic variability in the context of PD and treatment choices for optimizing glucose control. In this review, we have summarized the clinical considerations to guide and inform clinicians providing care for people with diabetes on PD.


Low carbohydrate diet rescued severe type 2 diabetes patient from insulin injection, a case report.
Fifty-six years old male type 2 diabetes patient with fasting blood sugar (FBS) 414 mg/d and hemoglobin A1c (HbA1c) 10.7% tried low carbohydrate diet (LCD) 30 days. After the diet, his FBS decreased to 174 mg/d and HbA1c was 7.7%. Therefore, he escaped from insulin injection and was able to treat with metformin.


Poly(hexamethylene biguanide) (PHMB) as high-efficiency antibacterial coating for titanium substrates.
Bacterial associated infection is a remaining urgent challenge in clinic application of metallic implants and devices. Here, we developed a new strategy to combat the bacterial associated infection of titanium alloy (TC4). Novel phosphonate/active ester block polymers (pDEMMP-b-pNHSMA) with identical phosphonate segments (DP = 29) as the metal anchorable ligand but varied active ester segments (DPs = 7, 29, and 64) as the conjugation site for poly(hexamethylene biguanide) (PHMB) were precisely prepared. Through a facile two-step process, the polymeric coating were successfully constructed on TC4 substrates as evidenced by water contact angle and XPS measurements. Through systematical in vitro antibacterial evaluations, robust relationship between the chemical structure of coating polymer and the antibacterial property endowed to the TC4 substrates has been established. Results showed that the block polymer, bearing an active ester segment of 64 repeat units, enabled dense packing of PHMB coating on the TC4 surface, which is able to kill 100% of both S. aureus and E. coli. that seeded without compromising the cytocompatibility of TC4 substrates. Furthermore, PHMB coating could significantly inhibit the colony of the bacteria and consequently reduce the bacterial associated inflammatory reaction as verified by a subcutaneous infection model on rat.


RRM adjacent TARDBP mutations disrupt RNA binding and enhance TDP-43 proteinopathy.
Amyotrophic lateral sclerosis (ALS) presents with focal muscle weakness due to motor neuron degeneration that becomes generalized, leading to death from respiratory failure within 3-5 years from symptom onset. Despite the heterogeneity of aetiology, TDP-43 proteinopathy is a common pathological feature that is observed in >95% of ALS and tau-negative frontotemporal dementia (FTD) cases. TDP-43 is a DNA/RNA-binding protein that in ALS and FTD translocates from being predominantly nuclear to form detergent-resistant, hyperphosphorylated aggregates in the cytoplasm of affected neurons and glia. Mutations in TARDBP account for 1-4% of all ALS cases and almost all arise in the low complexity C-terminal domain that does not affect RNA binding and processing. Here we report an ALS/FTD kindred with a novel K181E TDP-43 mutation that is located in close proximity to the RRM1 domain. To offer predictive gene testing to at-risk family members, we undertook a series of functional studies to characterize the properties of the mutation. Spectroscopy studies of the K181E protein revealed no evidence of significant misfolding. Although it is unable to bind to or splice RNA, it forms abundant aggregates in transfected cells. We extended our study to include other ALS-linked mutations adjacent to the RRM domains that also disrupt RNA binding and greatly enhance TDP-43 aggregation, forming detergent-resistant and hyperphosphorylated inclusions. Lastly, we demonstrate that K181E binds to, and sequesters, wild-type TDP-43 within nuclear and cytoplasmic inclusions. Thus, we demonstrate that TDP-43 mutations that disrupt RNA binding greatly enhance aggregation and are likely to be pathogenic as they promote wild-type TDP-43 to mislocalize and aggregate acting in a dominant-negative manner. This study highlights the importance of RNA binding to maintain TDP-43 solubility and the role of TDP-43 aggregation in disease pathogenesis.


[Influence of mutations in the suppressor of hairy wing and modifier of MDG4 loci on phenotype expression of super-unstable alleles at the yellow locus of Drosophila melanogaster].
A number of super-unstable systems were obtained earlier by the induction of P-M hybrid dysgenesis in the strains with a mobilized Stalker. One of the super-unstable mutations appeared at the yellow locus of Drosophila melanogaster on the background of preexisting y mutation. The latter had been induced by mdg4 insertion into the regulatory region of the yellow locus. The suppressor genes of Hairy wing and modifier of mdg4 are known to encode proteins which bind to the mdg4 enhancer and are involved in the control of its transcription. A large spectrum of yellow alleles was obtained in super-unstable systems which could be recognized by the intensity of pigmentation in twelve different areas of the cuticle. Combining of these alleles with su(Hw) and mod(mdg4) mutations followed by the study of phenotype changes demonstrated that the same regulatory protein may influence expression of the gene in opposite directions.


Radioactivity in trinitite six decades later.
The first nuclear explosion test, named the Trinity test, was conducted on July 16, 1945 near Alamogordo, New Mexico. In the tremendous heat of the explosion, the radioactive debris fused with the local soil into a glassy material named Trinitite. Selected Trinitite samples from ground zero (GZ) of the test site were investigated in detail for radioactivity. The techniques used included alpha spectrometry, high-efficiency gamma-ray spectrometry, and low-background beta counting, following the radiochemistry for selected radionuclides. Specific activities were determined for fission products (90Sr, 137Cs), activation products (60Co, 133Ba, 152Eu, 154Eu, 238Pu, 241Pu), and the remnants of the nuclear fuel (239Pu, 240Pu). Additionally, specific activities of three natural radionuclides (40K, 232Th, 238U) and their progeny were measured. The determined specific activities of radionuclides and their relationships are interpreted in the context of the fission process, chemical behavior of the elements, as well as the nuclear explosion phenomenology.


[Function, molecular structure and gene expression of insulin-like growth factors ].
In this short review, gene structure, transcriptional pattern, biological activity, specific receptors and binding proteins for insulin-like growth factors (IGF: IGF I and IGF II) are explained. In spite of their close similarity in both gene and peptide structures, their receptors are completely distinct from each other in molecular structure and in signal transduction. Expression of IGF function in vivo is strongly restricted by plasma binding proteins.


Contrasting environmental conditions precluded lower availability of Antarctic krill affecting breeding chinstrap penguins in the Antarctic Peninsula.
Dramatic decreases of chinstrap penguin populations across the Antarctic Peninsula (AP) are thought to be influenced by climate-driven changes affecting its main prey, the Antarctic krill, however, empirical evidence supporting such hypotheses are scarce. By coupling data on breeding chinstrap penguins, environmental remote sensing and estimates of krill acoustic density, we were able to demonstrate that penguins substantially increased their foraging effort in a year of low krill availability, with consequent reduction in breeding success. A winter of low sea ice cover followed by a summer/spring with stronger wind and lower marine productivity explained the lower and deeper krill availability. Our results highlight the importance of environmental variability on penguin populations, as variability is expected to increase under climate change, affecting foraging behaviour responses.


Gene within gene configuration and expression of the Drosophila melanogaster genes lethal(2) neighbour of tid [l(2)not] and lethal(2) relative of tid[l(2)rot].
In this paper, we describe the structure and temporal expression pattern of the Drosophila melanogaster genes l(2)not and l(2)rot located at locus 59F5 vis à vis the tumor suppressor gene l(2)tid described previously and exhibiting a gene within gene configuration. The l(2)not protein coding region, 1530 nt, is divided into two exons by an intron, 2645 nt, harboring the genes l(2)rot, co-transcribed from the same DNA strand, and l(2)tid, co-transcribed from the opposite DNA strand, located vis à vis. To determine proteins encoded by the genes described in this study polyclonal rabbit antibodies (Ab), anti-Not and anti-Rot, were generated. Immunostaining of developmental Western blots with the anti-Not Ab resulted in the identification of a 45-kDa protein, Not45, which is smaller than the Not56 protein predicted from the sequence. Its localization in endoplasmic reticulum (ER) was established by immunoelectron microscopy of Drosophila melanogaster Schneider 2 cells. Not45 shows significant homology to yeast ALG3 protein acting as a dolichol mannosyltransferase in the asparagine-linked glycosylation. It is synthesized ubiquitously throughout embryonic life. The protein predicted from the l(2)rot sequence, Rot57, shows a homology to the NS2B protein of the yellow fever virus1 (yefv1). The results of l(2)rot RNA analysis by developmental Northern blot and by in situ RNA localization, as well as the results of the protein analysis via Western blot and immunohistochemistry suggest that l(2)rot is transcribed but not translated. Since RNAs encoded by the genes l(2)tid and l(2)rot are complementary and l(2)rot is presumably not translated we performed preliminary experiments on the function of the l(2)rot RNA as a natural antisense RNA (asRNA) regulator of l(2)tid expression, expressed in the same temporal and spatial manner as the l(2)tid- and l(2)not RNA. l(2)tid knock-out by antisense RNA yielded late embryonic lethality resulting from multiple morphogenetic defects.


Post-bariatric Surgery Outcomes and Complications in Patients with Celiac Disease: a Matched Case-Control Study.
The shift towards an obese phenotype in celiac disease (CD) patients increases risk of morbidity and mortality. Bariatric surgery (BS) is the gold standard treatment for obesity. Few studies have explored the role of BS in patients with CD. This study aimed to assess the effectiveness and safety of BS in this population. This is a retrospective matched case-control (1:5) study of adult patients with confirmed CD who underwent BS at our institution from 1998 to 2018. Demographics, operative data, post-operative outcomes, complications, and nutritional parameters were collected. Seventy-eight patients (mostly Caucasian females) were included. Thirteen had confirmed CD and were compared with 65 controls. The most common type of BS was the RYGB. The percent of total body weight loss (%TWL) was similar for both groups at 6, 12, 18, 24, and 36 months, with the highest weight loss being at 12 months: %TWL 28.4 (20.2-38.4) for CD, n=13; vs. 29.1 (19.6-39.3) for non-CD, n=49; p=0.8. Obesity-associated co-morbidities greatly and comparably improved in both groups. Patients with CD had no complications after BS. Post-BS malodorous and oily stools were more common among patients with CD (23.1% vs. 4.6%, p=0.03). Micronutrient deficiencies were common and comparable among both groups with iron and vitamin D being the most common deficiencies. Gluten-free diet (GFD) non-adherence post-operatively was associated with a higher incidence of post-BS abdominal pain (60.0% vs. 0.0%, p=0.012). BS is safe and effective in patients with CD. Close monitoring is necessary to ensure compliance with GFD and vitamin supplementation. • Bariatric surgery leads to significant weight loss in celiac disease (CD) patients. • Mid- and long-term weight loss does not differ between patients with and without CD. • Obesity-associated co-morbidities significantly improve after surgery in CD patients. • The incidence of post-surgical complications is not higher in CD patients.


Differential inhibition of bacterial growth and hemolysin production by lincosamide antibiotics.
Lincomycin and clindamycin, at concentrations below those which partially inhibited bacterial growth, completely suppressed the production of streptolysin S. Chloramphenicol and erythromycin had no effect on hemolysin production.


A laboratory model for vascular microsurgery.
This article describes the microsurgical training model used at the Naval Regional Medical Center in San Diego, where first year residents spend two weeks of concentrated effort developing basic skills in microsurgery. This training is done outside the busy hospital environment so that no interruptions are encountered. Following this initial experience, weekly research sessions are continued to maintain proficiency and to develop new techniques. A description of the medical center's modifications of several previously used microsurgical techniques is presented. This model is economical and realistic for a student's initial exposure to microvascular surgery. It also serves as an expedient, yet challenging, means of maintaining proficiency when laboratory time is limited.


Fluid-structure interaction modeling of lactating breast: Newtonian vs. non-Newtonian milk.
Breastfeeding is a highly dynamic and complex mechanism. The suckling process by the infant involves compression and intra-oral vacuum pressure, leading to milk expression from breast. The accumulated milk from the nipple varies depending on the milk properties and transient flow rate during the suckling cycle. Rheological studies on raw human milk indicate that milk has a non-Newtonian shear-thinning flow behavior. This study aims to investigate the effect of non-Newtonian milk on flow behavior through the breast ductal system using fluid-structure interaction (FSI) simulation. The results of the non-Newtonian effects on flow velocity and the volumetric flow rate of expressed milk are presented. The results show that non-Newtonian Carreau model is promising for the simulation of human milk flow through the breast ductal systems. Also, the results show that the non-Newtonian effects on the milk flow behavior appear for 30-35% of the suckling cycle. Therefore, the Newtonian model is acceptable for the purpose of numerical simulation.


Skin manifestations in adults with gender dysphoria on hormone treatment in the outpatient clinic of Hospital de San José, Bogotá-Colombia.
Comprehensive healthcare for patients with gender dysphoria includes access to gender-affirming hormone therapy. It may cause cutaneous and adnexal side effects, which often affect quality of life, are underdiagnosed, and do not receive timely treatment. The literature on this subject is scarce. The main objective was to identify skin manifestations secondary to hormone treatment in adults with gender dysphoria. Observational, descriptive, cross-sectional study. A search was performed in the database of patients with gender dysphoria older than 18 years treated at the dermatology and/or endocrinology outpatient clinic of the Hospital de San José de Bogotá in the period 2015-2021. Medical records of patients on hormone therapy with skin manifestations were selected, while patients using other medications as possible triggers were excluded. In total, 85 patients were included, with a predominance of young transgender men (average age was 27 ± 9 years) in whom the main manifestation was acne (87%), followed by androgenetic alopecia and acanthosis nigricans. Transgender women presented more acne, androgenetic alopecia, hypertrichosis, seborrheic dermatitis, and melasma. The majority received treatment, mainly topical therapies. More than half of the patients were treated by the endocrinology and dermatology services. Skin manifestations in patients with gender dysphoria on hormone therapy are frequent, so they should be known and taken into account in the multidisciplinary approach to these patients, which should involve dermatologists. This is the first Colombian and Latin American study focused on documenting skin manifestations in patients with gender dysphoria undergoing hormone therapy.


Effects of continuous infusion of parathyroid hormone and parathyroid hormone-related peptide on rat bone in vivo: comparative study by histomorphometry.
We have investigated the actions of parathyroid hormone (PTH) and PTH-related peptide (PTHrP) on the bones of parathyroidectomized (PTX) rats by histomorphometric analysis. Miniosmotic pumps filled with either human PTH (hPTH)(1-34), hPTHrP(1-34) or vehicle were subcutaneously implanted on the backs of the rats. The peptides were continuously infused for 6 days at a rate of 15 nmole/kg/day. PTH and PTHrP exhibited similar hypercalcemic and hypophosphatemic actions on these PTX rats. No significant differences were noted in bone weight or calcium and phosphorus contents of the ashed bone among the 3 groups. By quantitative histomorphometric analysis, hPTH(1-34) and hPTHrP(1-34) were found similarly to enhance both bone formation and resorption. Peritrabecular fibrosis was observed only in the PTH-infused animals. PTHrP thus mimics the actions of PTH, but is not as effective in promoting mesenchymal cell proliferation along the bone trabeculae.


The estimated risk of miscarriage should be corrected for induced abortion rates.
We wanted to study to what extent induced abortion rates influence the age-dependent estimates of miscarriage risk. Register-based study. Linkage between hospital records and the Norwegian Population Registry. Included were all women 15-49 years of age in Oslo, Norway, who were pregnant during the three-year study period, 27,932 women. We estimated proportions with miscarriage according to maternal age with and without inclusion of women with induced abortion in the denominator. Proportions with miscarriage in each age group and odds ratios of miscarriage, with age group 25-29 years as the reference. When only women with pregnancies intended to term were included in the denominator, 9.4% of the women were estimated to have miscarriage as the only reproductive event during the three-year period and 6.8% had both miscarriage and delivery. The corresponding figures when also women with induced abortions were included in the denominator, were 7.0 and 5.0%. When taking induced abortion into account, the reduction in estimated proportion with miscarriage during the study period was greatest in the age groups with the highest induced abortion rates, women 15-19 years old (from 22.8 to 6.4%) and women 45-49 years old (from 57.9 to 41.0%). Estimates of age-dependent risk of miscarriage may be strongly influenced by induced abortion rates.


[Effective doses associated to the usual multimodal examinations in nuclear medicine].
To know the effective doses (E) that can be given in the common multimodal procedures in nuclear medicine, Single Photon Emission Tomography and Positron Emission Tomography, combined with Computed Tomography, SPECT/CT and PET/CT. Effective dose will be expressed according to Background Equivalent Radiation Time (BERT) and the contribution of the CT scan to the total dose will also be studied. The effective dose of each procedure has been calculated as the sum of the external radiation dose (CT part) and internal one (radiopharmaceutical administration). Data from each side were collected through extensive literature search. A range of minimum and maximum effective dose [E(min), E(max)] associated with each procedure is shown. These doses range from 0.5 to 49.1 mSv. BERT ranges from 2.6 months to 20.4 years. The CT contribution to the total effective dose varies from 2.1% to 93%, according to the image acquisition protocol and its purpose. Performing a diagnostic CT scan increases the dose up to a factor of 4.3 times compared to the dose used for CT acquisition for attenuation correction and anatomic localization. This article focus on the effective dose level that can be given in multimodal procedures, bearing in mind that the data are subject to constant changes and should be updated periodically. It is important to know the dosimetric impact when the CT scan is added. The application of optimized protocols according to the indication of the study reduces the patient's exposure without the loss of significant information.


Synthesis of the mitochondrial inner membrane in cultured Xenopus laevis oocytes.
The purpose of this study was to investigate the contribution of mitochondrial and cytoplasmic protein synthesis to the biogenesis of cytochrome oxidase (ferrocytochrome c:oxygen oxidoreductase EC 220.127.116.11) and rutamycin-sensitive adenosine triphosphatase (ATP phosphohydrolase EC 18.104.22.168) in cultured oocytes of the toad, Xenopus laevis. X. laevis cytochrome oxidase was purified over 23-fold with respect to specific activity and over 29-fold with respect to specific heme a content from oocyte submitochondrial particles. Polyacrylamide gel electrophoresis in the presence of sodium dodecyl sulfate separated the enzyme into six subunits with molecular weights of 44,000, 33,000, 23,000, 17,000, 12,000 and 9,500. the synthesis of the three larger subunits is sensitive to chloramphenicol (an inhibitor of mitochondrial protein synthesis), indicating that these subunits are made on mitochondrial ribosomes; the synthesis of the three smaller subunits is sensitive to cycloheximide (an inhibitor of cytoplasmic protein synthesis) and therefore occurs on cytoplasmic ribosomes. X. laevis rutamycin-sensitive ATPase, purified over 19-fold from oocyte submitochondrial pparticles, consists of 10 subunits with molecular weights of 56,000, 53,000, 41,000, 32,000, 29,000, 24,000, 21,000, 17,500 (2), and 11,500 on sodium dodecyl sulfate-polyacrylamide gels. The 29,000, 21,000, and one of the 17,500-dalton polypeptides are synthesized in the presence of cycloheximide and are, therefore, products of mitochondrial protein synthesis; the synthesis of the remaining seven subunits occurs in the presence of chloramphenicol, indicating that these subunits are made on cytoplasmic ribosomes. The synthesis of protein by mitochondria in cultured oocytes appears to be dependent upon cytoplasmic protein synthesis. In the presence of cycloheximide, the mitoribosomal synthesis of the subunits of cytochrome oxidase and rutamycin-sensitive ATPase is detectable only after a prior inhibition of mitochondrial protein synthesis by chloramphenicol. Oocyte mitochondrial ribosomes synthesize at least nine polypeptides after chloramphenicol treatment, three of which are components of neither cytochrome oxidase nor rutamycin-sensitive ATPase.


Photoaged microplastics induce neurotoxicity via oxidative stress and abnormal neurotransmission in zebrafish larvae (Danio rerio).
Microplastics (MPs) are ubiquitous environmental contaminants and cause neurotoxicity in various organisms. However, previous studies that analyzed the effects of MPs mainly focused on virgin polystyrene (V-PS) as representative models of MPs, and the mechanism underlying the neurotoxicity of photoaged polystyrene (P-PS) remains largely unknown. In this study, zebrafish (Danio rerio) were exposed to environmentally relevant concentrations (0.1-100 μg/L) of V-PS and P-PS(10 μm). The results indicated that UV radiation accelerated the aging process and changed physical and chemical properties of PS. Whereas exposure to both V-PS and P-PS at low concentrations (100 μg/L) significantly reduced the locomotor behavior of zebrafish larvae, P-PS caused more severe neurotoxicity compared to V-PS. The activity of antioxidant enzymes (SOD, CAT, and GST) and MDA content were significantly altered in zebrafish exposed to 10-100 μg/L of P-PS. Similarly, exposure to P-PS significantly increased neurotransmitter (5-HT, GABA, DA, and ACh) levels and activity of AChE, ChAT, and ChE. Star plots based on integrated biomarker response (IBR) values showed more incline toward neurotransmitter biomarkers in response to increasing P-PS concentration, and the behavioral parameters negatively correlated with the neurotransmitter biomarkers. Further investigations revealed that the expression of neurotransmission- (e.g., ache, drd3, 5th2c, and gat1) and oxidative stress- (e.g., cat1, sod1, gpx1a, and gstrl) related genes was significantly affected by PS in larval zebrafish. Thus, this study provides new insights on the potential risks of MPs into the environment.


Synthesis and biological activity of novel 4,4-difluorobenzazepine derivatives as non-peptide antagonists of the arginine vasopressin V1A receptor.
To find potent and selective antagonists of the arginine vasopressin (AVP) V1A receptor, optimization studies of compounds structurally related to (Z)-N-{4'-[(4,4-difluoro-5-carbamoylmethylidene-2,3,4,5-tetrahydro-1H-1-benzazepin-1-yl)carbonyl]phenyl}carboxamide were performed. The synthesis and pharmacological properties of these compounds are described. We first investigated the effect of the carboxamide moiety, and found that a 2-methylfuran-3-carbonyl group at this position increased V1A binding affinity and selectivity for the V1A receptor versus the V2 receptor. The amino group of the 5-carbamoylmethylidene moiety was also examined, and a 4-piperidinopiperidino group was found to be optimal at this position. The hemifumarate of compound 12l (YM218) was shown to exhibit potent binding affinity, V1A receptor selectivity, and in vivo antagonist activity.


Dissemination of IncFII(K)-type plasmids in multiresistant CTX-M-15-producing Enterobacteriaceae isolates from children in hospital paediatric oncology wards.
In this study, extended-spectrum β-lactamase (ESBL)-producing Enterobacteriaceae isolates in children with malignancies hospitalised at a paediatric oncology department in the Czech Republic were investigated. From June 2009 to January 2010, a total of 50 ESBL-producing faecal isolates of Enterobacteriaceae were obtained from 28 patients. These isolates were characterised with regard to ESBL enzymes, plasmid-mediated quinolone resistance genes, multilocus sequence typing (MLST) and plasmids conferring resistance to cephalosporins and fluoroquinolones. ESBL-producing isolates included Klebsiella pneumoniae (n=36), Escherichia coli (n=7), Klebsiella oxytoca (n=3), Enterobacter cloacae (n=2) and Citrobacter freundii (n=2). Klebsiella pneumoniae isolates belonged to 7 MLST types, including sequence types ST280, ST321, ST323 and ST416 as well as the novel types ST626, ST627 and ST628. The multiresistant epidemic clone E. coli B2-O25b-ST131 was detected in one patient. The gene bla(CTX-M-15) was found on large conjugative IncFII(K) plasmids along with bla(TEM-1), bla(OXA-1), qnrB1, aac(6')-Ib-cr, strA, sul2, aac(3')-II and tet(A) genes in most isolates. Dissemination of IncFII(K) plasmids among various Enterobacteriaceae isolates was considered an important aspect of nosocomial colonisation in the wards by Enterobacteriaceae species producing ESBLs. This is the first study documenting multiple antibiotic resistance elements, including qnr genes, in IncFII(K) plasmids in various bacterial species isolated in a single hospital department. The results highlight the evolution of IncFII(K) plasmids into new variants containing novel antibiotic resistance elements and their important role in spreading ESBL-producing bacteria among hospitalised patients.


Carbon and nitrogen isotopic compositions of particulate organic matter in the upwelling zone off the east coast of Hainan Island, China.
The isotopic compositions (δ13C and δ15N) and C/N ratios of suspended particulate organic matter (POM) were investigated off the east coast of Hainan Island in the South China Sea during summer. Coastal upwelling influenced the nearshore stations of transects S2 and S3, and higher δ13C and δ15N values suggested that coastal upwelling played a significant role in determining the POM sources. The POM at the nearshore area of transect S1 was controlled by the coastal current and freshwater discharge. Additionally, organic matter may be transported to the offshore area via tidal movements in transects S1 and S3. Based on the stable isotope analysis in an R model, the marine organic matter contribution in the upwelling area (19%) was higher than that in the other areas (transect S1 and the offshore area) (7%). The δ13C and δ15N values and C/N ratios reflect the carbon and nitrogen sources and their cycling in the upwelling zone off the east coast of Hainan Island.


Frequencies of GB virus C/hepatitis G virus genomes and of specific antibodies in German risk and non-risk populations.
The prevalence of the new flavivirus GB virus C/hepatitis virus G (GBV-C/HGV) in different German populations was investigated by detection of viral genomes and anti-E2 antibodies. While blood donors had an overall prevalence of 10.4% there were increased rates for hemophiliacs (54.7%), hemodialysis patients (30.2%), male homosexuals (30.2%) and intravenous drug users (74.4). Most GBV-C/HGV positive samples were either viral genome positive or antibody positive, exclusively. Samples with the rare constellation "positive for both GBV-C/HGV genome and specific antibody" originated in almost all cases from patients who were additionally infected with HIV or HCV. Probable transmission of GBV-C/HGV by PCR-positive blood transfusions was observed in 5 of 6 cases approximately six months after transfusion.


Danazol suppression of luteinizing hormone secretion: a role for danazol-androgen receptor interaction within the brain-pituitary complex.
We investigated the relationship between intracellular androgen and estrogen receptors within the brain-pituitary complex and danazol suppression of luteinizing hormone secretion in castrated rats. Danazol did not influence 3H-estradiol-estrogen receptor interactions within the brain and pituitary but did suppress 3H-dihydrotestosterone-androgen receptor interactions within this neuroendocrine complex. To study the significance of these danazol-androgen receptor interactions, the drug was administered to androgen receptor-deficient pseudohermaphrodite or flutamide-treated male rats. In both instances, danazol suppression of luteinizing hormone release was markedly attenuated or absent. These findings point to danazol-androgen receptor interactions within the brain-pituitary complex as a key step in this drug's antigonadotropic actions. Whether the active drug form is danazol itself or a metabolite is subject to further study.


Stable Chronic Obstructive Pulmonary Disease (COPD) Management Under a Tiered Medical System in China.
Early diagnosis and proper management of a large number of chronic obstructive pulmonary disease (COPD) patients are great challenges for the Chinese health care system. Although tiered medical services have been promoted by the Chinese government since 2015, they have not been ideally implemented for COPD diagnosis and management. We designed a cross-sectional study. Eligible COPD patients (n = 648) and physicians (n = 161) were consecutively recruited from 8 hospitals in different tiers in China. COPD characteristics and treatments were compared among hospitals in different tiers. Multivariate logistic regression was performed to identify risk factors associated with airflow limitation, symptoms and acute exacerbation. The PFT rate at first diagnosis was 99%, 69.4% and 29.9% in teaching, second-tier and community hospitals (P < 0.001). Only approximately 10.9%, 1.7% and 9.6% and 21.8%, 6.9% and 32% of COPD patients received influenza or pneumococcal vaccines (P < 0.001). The proportion of patients who did not use inhaled drugs or had irregular inhalation was 2%, 24.6% and 78.8% (P < 0.001). Education level (RR-1 = -41.26%, P = 0.007), FEV1%pred (RR-1 = -2.76%, P < 0.001), and influenza vaccination in the last year (RR-1 = -64.53%, P = 0.006) were all negatively correlated with COPD acute exacerbation (AE). COPD duration (RR-1 = 131.73%, P = 0.009), AE (RR-1 = 151.39%, P < 0.001), and COPD Assessment Test (CAT) scores (RR-1 = 3.82%, P = 0.019) were all positively correlated with COPD airflow limitation severity. Differences exist in the diagnosis, treatment and management of COPD among different tiers of hospitals in China. Teaching hospitals can manage COPD patients relatively well. There are still some gaps compared with developed countries.


Pediatric Advanced Life Support: Part II. Fluid therapy, medications and dysrhythmias.
The first part of this two-part article discussed the equipment needed for pediatric resuscitations and the techniques used for cardiopulmonary assessment, airway securance, circulatory maintenance and intravascular access. In this second part, additional life support measures are reviewed, including fluid therapy, resuscitation medications and the management of cardiac rhythm disturbances.


Determinants and patient-reported long-term outcomes of physician empathy in oncology: a structural equation modelling approach.
The aim of the present cross-sectional study was to explore patient- and physician-specific determinants of physician empathy (PE) and to analyse the influence of PE on patient-reported long-term outcomes in German cancer patients. A postal survey was administered to 710 cancer patients, who had been inpatients at the University Hospital Cologne (response rate 49.5%). PE was measured with the German translation of the consultation and relational empathy (CARE) measure, and patient-reported long-term outcomes were assessed using the major (ICD-10) depression inventory (MDI) and the EORTC quality of life (Qol) questionnaire QLQ-C30. Hypotheses were tested by structural equation modelling. PE had (a) a moderate indirect effect on "depression" and a smaller indirect effect on "socio-emotional-cognitive Qol" by affecting "desire for more information from the physician regarding findings and treatment options" and (b) a moderate indirect effect on "socio-emotional-cognitive Qol" and a smaller effect on "depression" via "desire for more information about health promotion". The determinant with the greatest importance was "patient-perceived general busyness of hospital staff": it had a strong negative influence on PE, indirectly influencing "desire for more information from the physician regarding findings and treatment options" and also patients' "depression". PE seems to be an important pre-requisite for information giving by physicians and through this pathway having a preventive effect on depression and improving Qol. Conversely, physicians' stress negatively influences these relationships. The research findings suggest that reducing physicians' stress at the organizational and individual may be required to enhance patient-physician communication. Empathy, as an outcome-relevant professional competence needs to be assessed and developed more intensively in medical students and physicians.


Translational control by helicases during cellular stress.
Stress is inevitable, so all organisms have developed response mechanisms to allow for their survival during times of stress. Regulation of gene expression is a critical part of these responses, which allows for the appropriate cohort of proteins to be produced to counter the stress while downregulating others in order to conserve resources. Translation is both highly energy intensive and able to rapidly shift the proteome, thus making it a key target for regulation during stress. Numerous stress pathways converge on translation, and examining the regulatory mechanisms that underlie these pathways is essential for understanding the initial and long-term effects of stress on cells. A number of RNA helicases, including eIF4A, Ded1/DDX3X, and Dhh1/DDX6, have been previously linked to translation, and given their ability to dramatically alter RNA-protein interactions, they are well-positioned to play critical roles in translation regulation during stress. Therefore, assessing the role of helicases in these conditions is vital to the overall understanding of stress. Outlined below are key assays focusing on two areas: assessing cellular phenotypes in growth and survival during stress conditions, and analyzing cellular translation in the presence and absence of stress. The combination of these two approaches will begin to establish the function(s) of a given helicase in the overall stress response.


A case of bilateral sclerouveitis with secondary glaucoma right eye.
Infectious scleritis is rare and most commonly herpetic in origin. We report an unusual bilateral subacute presentation of scleritis with uveitis and glaucoma which responded to treatment with acyclovir. A 47-year-old male coast guard personnel presented with 2 months history of bilateral red eye. He was initially managed elsewhere as conjunctivitis, and on examination had bilateral diffuse redness of the eyes persisting with phenylephrine, with scleral edema and mild globe tenderness. Investigations for underlying autoimmune systemic illness were normal. He showed inadequate response to topical steroids and cycloplegics and developed uveitis and glaucoma while on steroids. He was then given tablet acyclovir with antiglaucoma topical medications with which he showed rapid response and complete resolution. The case is being reported for highlighting this unusual bilateral presentation of scleritis with uveitis and glaucoma with possible viral etiology.


[Asphyxia of the newborn delivered by elective cesarean section and gastroesophageal reflex and aspiration].
To analyse the causes, clinical manifestation, management and prevention of gastroesophageal reflex and aspiration (GER-A) in the infants delivered by cesarean section. The clinical data of 6 infants with asphyxia resulted from GER-A, delivered by elective cesarean section were retrospectively analysed. GER-A occurred during the period from 5 to 12 minute after birth, and its main clinical manifestation was airway obstruction. Among the patients, four cases recovered completely after treatment, one was diagnosed as having aspiration pneumonia and the other died of severe bronchospasm. The occurrence of this complication was probably related to the mode of delivery by cesarean section, the underdevelopment of effective barrier for preventing GER-A at very early stage and the inadequate management of the newborn. It is suggested that the prevention and proper treatment of GER-A was important in lowering neonatal morbidity and mortality.


Food purchase decision-making typologies of women with non-insulin-dependent diabetes mellitus.
Food selection is a key factor in the nutritional management of diabetes mellitus. Since up to 80% of food purchase decisions are made at the supermarket, the purpose of this study was to identify the criteria which influence point-of-purchase decision-making in women with NIDDM aged 40-60 years. A qualitative approach with individual interviews and in-store observations was used. Analysis of the interviews identified four decision-making typologies based on the extent nutrition, price and family needs were emphasized. The four typologies included (1) the Overloaded Shopper, (2) the Budget Shopper, (3) the Nutrition Savvy Shopper, and (4) the Out-of-Touch Shopper. Cluster analysis confirmed the typologies for 71% of the sample. Educators should classify shoppers according to a typology to determine their clients' personal needs and interests. Then, educators can tailor the educational or counseling message to meet those specific needs.


The molecular arms race between African trypanosomes and humans.
Humans can survive bloodstream infection by African trypanosomes, owing to the activity of serum complexes that have efficient trypanosome-killing ability. The two trypanosome subspecies that are responsible for human sleeping sickness--Trypanosoma brucei rhodesiense and Trypanosoma brucei gambiense--can evade this defence mechanism by expressing distinct resistance proteins. In turn, sequence variation in the gene that encodes the trypanosome-killing component in human serum has enabled populations in western Africa to restore resistance to T. b. rhodesiense, at the expense of the high probability of developing kidney sclerosis. These findings highlight the importance of resistance to trypanosomes in human evolution.


Laminar distribution of receptors in monkey (Macaca fascicularis) geniculostriate system.
We have examined the laminar distributions of eight types of receptor in the primary visual cortex (area 17) and the lateral geniculate nucleus (LGN) of the macaque monkey. The receptor populations and subpopulations examined included those selective for gamma-aminobutyric acid (GABA) (using [3H]-muscimol as ligand), L-glutamate-related receptors (using [3H]-L-glutamate and [3H]-AMPA), muscarinic acetylcholine (using [3H]-quinuclidinyl benzilate--QNB and [3H]-N-methyl scopolamine--NMS), cholecystokinin (CCK) (using [3H] pentagastrin), benzodiazepine (using [3H]-flunitrazepam), and adenosine (using [3H]-cyclohexyladenosine--CHA). Each of the receptors examined exhibited characteristic and differing laminar patterns of binding in the striate cortex. Perhaps reflecting the high density of cell bodies and synapses in layer 4C, most receptors, except those labelled by [3H]-L-glutamate or [3H]-AMPA, showed dense concentrations in this layer. Layers 4B and 5, which contain relatively few cell bodies and heavy myelin concentrations, were in general lightly labelled. Layer 6 showed relatively heavy labelling when [3H]-AMPA (quisqualate) or [3H]-pentagastrin (CCK) were used as ligands. The superficial layers of the cortex were zones of relative concentration of GABA, benzodiazepine, acetylcholine, glutamate-related, and adenosine receptors. In general, the binding patterns resembled those previously described for cat visual cortex, but there were also some clear differences. The distributions of all of these receptors likely reflect the differential input substances to different laminae of the visual cortex. Of the receptors examined, only those for GABA, benzodiazepine, and acetylcholine were found in substantial concentration in the LGN. Of these, GABA and benzodiazepine receptors showed especially dense binding in the magnocellular layers of the LGN compared to the parvicellular layers.


Mechanism and biological significance of constitutive expression of MGSA/GRO chemokines in malignant melanoma tumor progression.
By reverse transcriptase-polymerase chain reaction, enzyme-linked immunosorbent assay, and immunohistochemistry, MGSA-alpha, -beta, -gamma, and CXCR2 mRNA expression and proteins are detected in 7 out of 10 human melanoma lesions. The biological consequence of constitutive expression of the MGSA/GRO chemokine in immortalized melanocytes was tested in SCID and nude mouse models. Continuous expression of MGSA/GRO-alpha, -beta, or -gamma in immortalized melan-a mouse melanocytes results in nearly 100% tumor formation for each of the clones tested, whereas clones expressing only the neomycin resistance vector form tumors <10% of the time. Moreover, antibodies to the MGSA/GRO proteins slow or inhibit the formation of tumors in the SCID mouse model and block the angiogenic response to conditioned medium from the tumor-producing clones. Transcription of the MGSA/GRO chemokines is regulated by an enhancesome-like complex comprised of the nuclear factor-kappaB (NF-kappaB), HMG(I)Y, IUR, and Sp1 elements. In Hs294T melanoma cells the half life of the IKB protein is shortened in comparison to normal retinal epithelial cells, facilitating the endogenous nuclear localization of NF-kappaB. We propose that this endogenous nuclear NF-kappaB, working in concert with the 115-kDa IUR-binding factor, promotes constitutive expression of MGSA/GRO genes.


An electron paramagnetic resonance study of free radicals in cells.
An electron paramagnetic resonance study was performed on cell lines of the following strains: HeLa, 37RC, L, FLC, NRK/RSV, 3T3/SV40. Unsynchronized and synchronized HeLa cells were studied with particular attention paid to the relation between growth and free radical concentration. Free radical levels were shown to be a function of the growth stage and different phases of the cell cycle.


Comparison of effectiveness of frequently and infrequently used homeopathic medicines.
Patients treated with homeopathy may respond to infrequently used or even 'new' medicines. But does the introduction of an unlimited number of (new) medicines improve homeopathy? Do new medicines solve old problems? 1. Consensus meetings to evaluate best cases. 2. Patient outcome study in 10 Dutch practices. Good cases are scarce for many medicines, random variance is an important source of uncertainty. 50 Medicines are responsible for 72% of all successful prescriptions. There is no difference in effectiveness of frequently and less frequently used medicines. Confirmation bias is found for a few well-known symptom-medicine combinations. 'New' and infrequently medicines are as effective as 'old' frequently used medicines. Improving the use of frequently used medicines is more effective in improving results than seldom used medicines. Large numbers are required and old mistakes should be avoided developing new medicines. A research agenda for improving the use of homeopathic medicines is imperative.


Facilitated temporal summation of pain at spinal level in Parkinson's disease.
Pain is one of the major nonmotor symptoms of Parkinson's disease. We hypothesized that Parkinson's disease patients could show an early diffuse abnormal processing of the nociceptive inputs also in the absence of clinical pain syndrome and that this could represent the physiopathological substrate to explain the high incidence of diffuse pain symptoms. We used the temporal summation threshold of the nociceptive withdrawal reflex and the related pain sensation to evaluate the facilitation in pain processing at spinal level. Fifteen (7 Women; 8 Men; mean age 63.0 ± 9.1) Parkinson's disease patients without clinical pain and 12 (6 Women, 6 Men; mean age 61.2 ± 4.2) healthy subjects were recruited. Parkinson's disease group has been subdivided into two subgroups, 7 early-stage Parkinson's disease patients with unilateral signs (Hoehn and Yahr stage 1) and 8 patients in a more advanced stage of the disease showing bilateral parkinsonian signs (Hoehn and Yahr stages 2 and 2.5), both "on" and "off" treatments with levodopa. A significant facilitation in temporal summation of pain (reduced temporal summation threshold and increased painful sensation) was found in Parkinson's disease patients when compared with controls. This facilitation is more evident in Parkinson's disease with bilateral signs and on the side more affected in Parkinson's disease with unilateral signs. Levodopa administration failed to significantly modify the neurophysiological abnormalities; however, a slight improvement has been detected. The increased gain in pain processing at spinal level in Parkinson's disease patients could be a consequence of the degenerative phenomena involving supraspinal projections implicated in the modulation of pain processing and could make Parkinson's disease patients more predisposed to develop a pain condition.


Cyclooxygenase-2 Directs Microglial Activation-Mediated Inflammation and Oxidative Stress Leading to Intrinsic Apoptosis in Zn-Induced Parkinsonism.
Inflammation is decisive in zinc (Zn)-induced nigrostriatal dopaminergic neurodegeneration; however, the contribution of cyclooxygenase-2 (COX-2) is not yet known. The present study aimed to explore the role of COX-2 in Zn-induced Parkinsonism and its association with the microglial activation. Male Wistar rats were treated intraperitoneally (i.p.) with Zn as zinc sulphate (20 mg/kg) along with respective controls for 2-12 weeks. In a few sets, animals were also treated with/without celecoxcib (CXB, 20 mg/kg, i.p.), a selective COX-2 inhibitor. Indexes of the nigrostriatal neurodegeneration, oxidative stress, inflammation and apoptosis were measured in the animals/nigrostriatal tissue. Zn induced time-dependent increase in the expression of COX-2 while COX-1 expression was unaltered. Zn reduced the neurobehavioral activities, striatal dopamine content, tyrosine hydroxylase (TH) expression and number of dopaminergic neurons. While oxidative stress; microglial activation; expression of microglial cell surface marker-CD11b; cytochrome c release; caspase-9/3 activation; level of pro-inflammatory cytokines, such as TNF-α, IL-1β and IL-6 and Bcl-2-associated protein x (Bax) translocation from the cytosol to mitochondria were induced in the Zn-treated group, expression of B-cell lymphoma-2 (Bcl-2) was found to be reduced. CXB significantly attenuated Zn-induced increase in COX-2 expression and restored TH-expression, dopamine content, level of inflammatory cytokines and neurobehavioral indexes towards normalcy. Moreover, CXB also attenuated Zn-induced increase in microglial activation, oxidative stress and apoptotic markers towards normal levels. Results of the study thus demonstrate that COX-2 induces microglial activation that provokes the release of inflammatory mediators, which in turn augments oxidative stress and intrinsic apoptosis leading to dopaminergic neurodegeneration in Zn-induced Parkinsonism.


Identification and interaction mechanism of protein corona on silver nanoparticles with different sizes and the cellular responses.
With the potential biomedical applications of nanomaterials such as silver nanoparticles (SNPs), nanotoxicity concerns are growing, and the importance of NP and protein interactions is far from being addressed enough. Here, we identified the major binding protein on SNPs in blood as human serum albumin (HSA) using polyacrylamide gel electrophoresis and liquid chromatography-mass spectrometry/mass spectrometry. By comparing with the previous methods, we emphasized surface area concentration as a new dose metric to address the importance of NP curvature. SNPs interacted with cysteine and cystine, disrupting the secondary structure and conformation of HSA, and this tendency became stronger on small SNPs than large ones. The protein corona significantly alleviated the toxicity and decreased SNPs' internalization in a particle size-dependent manner, where more significant inhibition effects occurred on larger particles at the same area concentration. These findings may shed light on nanotoxicity and also the design of safe nanomaterials by a comprehensive preconsideration of the metrological method.


Diffusive permeability of rabbit peritoneum for small solutes in vitro: effect of gentamicin and insulin.
The purpose of the in vitro study was to compare the diffusive transport of creatinine, uric acid and glucose, directed from the interstitial (I) to the mesothelial (M) side of the peritoneum and in the opposite direction, before (15-60min) and after (75-120min) application of gentamicin and insulin. The experiments were undertaken on the rabbit parietal peritoneum in a modified Ussing chamber. A mathematical model was used to calculate a diffusive permeability coefficient P (in cm/s) and a diffusive mass transport coefficient KH (in mL/min). In the basic experimental series without drugs, the dynamics of peritoneal transport for examined solutes remained constant and there were no differences between transport directions (I--> M and M--> I). Mean values of P+/-SEM (KH+/-SEM) were 3.06+/-0.32 (321.1+/-33.5), 2.09+/-0.29 (219.2+/-30.7) and 2.73+/-0.47 (286.7+/-49.6) for creatinine, uric acid and glucose, respectively. The introduction of gentamicin decreased glucose transport directed from the mesothelial to the interstitial side of the membrane by about 12%. After insulin application we observed the increase of creatinine and glucose peritoneal transport rate. For creatinine, the above augmentation was by about 31% for I--> M and 83% for M--> I direction. In these conditions, the glucose transport directed from the interstitial to the mesothelial side of membrane increased by about 24%. Generally, in vitro gentamicin decreases, but insulin increases the diffusive permeability of the peritoneum for some small solutes. We suppose that these findings may be important for the efficiency of peritoneal dialysis.