[The role of metformin in the treatment of dermatological diseases: A narrative review].
To review and discuss the current evidence of the use of metformin as a therapeutic tool in frequent skin diseases. Original article. Qualitative research. Narrative review. Aragon and Murcia, Spain. Resident Physicians. Dermatology and Primary Health Care. A narrative review has been carried out using the PubMed bibliographic database, being the search date the 27th of January of 2022. Metformin has proven to be effective in the treatment of inflammatory skin diseases such as acne, hidradenitis suppurativa, psoriasis and allergic contact dermatitis. It has also shown antitumor properties regarding basal cell carcinoma, squamous cell carcinoma and melanoma. Additionally, beneficial effects of adjuvant treatment with metformin have been described in patients with basal cell carcinoma receiving photodynamic therapy. In patients with endocrinology-related dermatosis such as hirsutism, acanthosis nigricans and eruptive xanthomas, treatment with metformin has demonstrated therapeutic effectiveness. Topical treatment with metformin has also been useful in the treatment of melasma. Finally, it has been proposed as a drug with anti-aging and wound-healing promoting properties. Severe adverse effects have not been observed for any of the previously described indications, being this a well-tolerated treatment. Metformin is an effective and safe adjuvant in the therapeutic scheme of various inflammatory dermatoses, skin neoplasms, endocrinology-related dermatosis, melasma, skin aging and wound healing processes.


Studies of the biosynthesis of a soluble precursor of elastin.
A salt soluble protein, isolated from the aortas of copper-deficient swine, has been characterized as a precursor of elastin by its amino acid composition and its incorporation into insoluble aortic elastin in vitro.


Involvement of dipeptidyl peptidase IV in immune complex-mediated glomerulonephritis.
Dipeptidyl peptidase IV (DPPIV) is widely expressed in many tissues; however, its precise biological function is poorly understood. One of its possible physiologic roles is an involvement in the immune system, which plays a pivotal role in the pathogenesis of glomerulonephritis. The present study focused on the involvement of DPPIV in immune complex-mediated glomerulonephritis. Experimental nephritis was induced by anti-Thy-1.1 monoclonal antibody E30 using Wistar or F344 rats. The application of a new monoclonal antibody against DPPIV, F16, completely suppressed E30-induced proteinuria and mesangial proliferation in Wistar rats, whereas these preventive effects of F16 were not observed in F344 rats, which spontaneously lack DPPIV protein. Treatment with F16 inhibited glomerular deposition of complement C3 and complement C4 after the binding of E30 to the mesangial cell surface. Because the preventive effect of F16 was attributable to suppression of the complement cascade, we examined its influences on complement-dependent mesangial cell lysis in vitro. We discovered that the complement cascade was markedly inactivated in F16-treated Wistar rat serum but not in F16-treated F344 rats. These results indicate that DPPIV may play a somewhat crucial role in regulating the complement cascade and that inhibition of DPPIV may serve as a new target for preventing complement-dependent tissue injury.


Electrophysiological estimates of the time course of tonal and orthographic encoding in Chinese speech production.
Recent electrophysiological studies have investigated the time course of semantic, syntactic, and phonological encoding in European language spoken production, such as English or Dutch. The present study investigated the time course of tonal and orthographic encoding during Chinese word production. Participants were shown pictures and carried out a dual-choice Go/noGo decision based on tonal information (whether a picture name was tone 1 or 2, or tone 3 or 4) or orthographic information (whether or not the picture name was written with a left-right structure character). Analyses of N200 effects and LRPs (lateralized readiness potentials) indicated that tonal information was retrieved prior to orthographic information. These results imply that orthographic codes are unlikely to contribute to phonological encoding in spoken word production. Furthermore, a late effect for the N200 in the Go/noGo=tone condition was observed, which may be related to internal self-monitoring of suprasegmental information.


Characterization of the Corynebacterium glutamicum dehydroshikimate dehydratase QsuB and its potential for microbial production of protocatechuic acid.
The dehydroshikimate dehydratase (DSD) from Corynebacterium glutamicum encoded by the qsuB gene is related to the previously described QuiC1 protein (39.9% identity) from Pseudomonas putida. Both QuiC1 and QsuB are two-domain bacterial DSDs. The N-terminal domain provides dehydratase activity, while the C-terminal domain has sequence identity with 4-hydroxyphenylpyruvate dioxygenase. Here, the QsuB protein and its N-terminal domain (N-QsuB) were expressed in the T7 system, purified and characterized. QsuB was present mainly in octameric form (60%), while N-QsuB had a predominantly monomeric structure (80%) in aqueous buffer. Both proteins possessed DSD activity with one of the following cofactors (listed in the order of decreasing activity): Co2+, Mg2+, Mn2+. The Km and kcat values for the QsuB enzyme (Km ~ 1 mM, kcat ~ 61 s-1) were two and three times higher than those for N-QsuB. 3,4-DHBA inhibited QsuB (Ki ~ 0.38 mM, Ki' ~ 0.96 mM) and N-QsuB (Ki ~ 0.69 mM) enzymes via mixed and noncompetitive inhibition mechanism, respectively. E. coli MG1655ΔaroEPlac‒qsuB strain produced three times more 3,4-DHBA from glucose in test tube fermentation than the MG1655ΔaroEPlac‒n-qsuB strain. The C-terminal domain activity towards 3,4-DHBA was not established in vitro. This domain was proposed to promote protein oligomerization for maintaining structural stability of the enzyme. The dimer formation of QsuB protein was more predictable (ΔG = ‒15.8 kcal/mol) than the dimerization of its truncated version N-QsuB (ΔG = ‒0.4 kcal/mol).


De novo assembly of the Mylia taylorii transcriptome and identification of sesquiterpene synthases.
Mylia taylorii is an ancient nonseed land plant that accumulates various sesquiterpenes with insecticidal and antibacterial activities. Recently, microbial-type sesquiterpene synthases (STSs) with atypical aspartate-rich metal ion binding motifs have been identified in some liverworts. Here, transcriptome analysis of M. taylorii was performed to identify M. taylorii sesquiterpene synthases (MtSTSs) that are potentially involved in sesquiterpene biosynthesis and diversity. A total of 255,669 unigenes were obtained with an average length of 963 bp in the transcriptome data of M. taylorii, among which 148,093 (57.92%) unigenes had BLAST results. Forty-eight unigenes were related to the sesquiterpene backbone biosynthesis according to KEGG annotation. In addition, MtSTS1, MtSTS2 and MtSTS3 identified from putative MtSTSs display sesquiterpene catalytic activities on the basis of functional characterizations in yeast. Interestingly, MtSTSs exhibit a noncanonical metal ion binding motif and the structural composition of a single α-domain, which are features of microbial STSs instead of archetypical plant STSs. This study revealed new microbial-type STS members of nonseed plants, and functionally identified that MtSTSs may contribute to the investigation of the biosynthesis and biological role of sesquiterpenes in M. taylorii.


The Application of Nano-Sized Zero-Valent Iron for In Situ Remediation of Chlorinated Ethylenes in Groundwater: A Field Case Study.
In this paper, the authors present the pilot in situ application of nano zero-valent iron (nZVI) for effective remediation of groundwater in an industrial area contaminated by chlorinated ethylenes (CEs), which create a significant group of global environmental contaminants. This work covers the entire 1-year remediation process, including systematic laboratory tests and field application techniques for nZVI. The application was carried out in the area of a metal fabrication industrial facility in the Czech Republic. Contamination of CEs in this area is a consequence of old ecological loads. The entire remediation process contained the following steps: monitoring of the area, selection of the most relevant hot spot, selection of the most appropriate application borehole, systematic laboratory tests, application of nZVI, and postapplication monitoring. Ten kilograms of nZVI were applied as a water suspension into the selected borehole. Significant decreases in concentrations of selected contaminants were observed in the first month after application. The reaction in the borehole was completed within approximately 5 to 6 months after the application and during this period almost 50% elimination of contamination was achieved.


[The efficacy of treatment using expanded programs of megavolt irradiation and polychemotherapy in patients with lymphogranulomatosis in stages I, II and IIIA].
The authors relate the results of multimodality treatment of 90 patients with stages I-IIIA lymphogranulomatosis. The total survival amounted to 86.4 +/- 3.8% within the time exceeding 7 years. The efficacy of the treatment of patients with lymphogranulomatosis of different stages by using multi-beam and broad-beam radiation is under comparison. The data obtained indicate that total radiation of lymph nodes in patients with stage II lymphogranulomatosis is not advisable.


Microbial mutagenicity of 3- and 4-ring polycyclic aromatic sulfur heterocycles.
The stable isomers of 3- and 4-ring polycyclic aromatic sulfur heterocycles were tested for mutagenicity in the Ames standard plate incorporation test and a liquid pre-incubation modification of the Ames test. Of the 4 three-ring compounds tested, only naphtho[1,2-b]thiophene was mutagenic. Of the four-ring compounds, 7 of 13 were mutagenic in the standard Ames or pre-incubation Ames test. The highest activity for the 4-ring compounds was observed for phenanthrol[3,4-b]thiophene, a compound of approximately the same mutagenic potency in the Ames test as benzo[a]pyrene. The other active 4-ring compounds were of considerable less mutagenic potency than phenanthrol[3,4-b]thiophene. Mutagenicity for two of the 4-ring aromatic thiophenes could only be detected in the liquid pre-incubation Ames test. Salmonella typhimurium TA100 was the most sensitive strain to mutagenesis by these compounds, followed by TA98. All mutagenesis was indirect, requiring metabolic activation.


Validation of plasma D-dimer in Chinese patients with acute non-ST segment elevation myocardial infarction.
To analyze the predictive values of D-dimer in Chinese patients with non-ST-segment elevation myocardial infarction (NSTEMI). We retrospectively retrieved consecutive patients hospitalized due to acute NSTEMI from January 2015 to December 2018 from the Electronic Medical Record (EMR) library. Clinical and follow-up data were collected. The primary endpoint was major adverse composite cardiovascular events (MACEs), such as all-cause death, non-fatal myocardial infarction, and non-fatal stroke. The secondary endpoints included all-cause death, non-fatal myocardial infarction, non-fatal stroke, heart failure, and severe arrhythmias. The Cox regression model was used to evaluate the association between risk factors and clinical outcomes in Chinese patients with NSTEMI. A total of 673 patients were included; the median age was 64.0 (53.0-75.0) years old and 76.2% were men. Patients with higher D-dimer levels were more often women, older, had a higher Charlson Comorbidity Index, and had a higher incidence of MACEs (59.9 vs. control 9.0%; p < 0.001) and all-cause death (49.1 vs. control 2.2%; p < 0.001). The multivariate Cox analysis suggested that the D-dimer level was an independent predictor of MACEs (hazard ratio [HR]: 1.069, 95% CI: 1.010-1.132, p = 0.021). The receiver operating characteristic (ROC) analysis suggested that D-dimer levels were better than the Charlson Comorbidity Index in all-cause death. In Chinese patients with acute NSTEMI, higher D-dimer levels on admission suggest a poor long-term prognosis.


Identifying Diagnostic and Prognostic Differentially Expressed Genes of Gastric Cancer Based on Bioinformatics Analyses of RNA-seq Data.
Background: The abnormal expression of genes in serum may be associated with early diagnosis of patients with malignant tumors. This study was designed to screen for significantly differentially expressed genes (DEGs) that may be associated with gastric cancer using bioinformatic methods. Methods: RNA-seq data from gastric cancers were downloaded from the TCGA and GEO databases, and 1903 secretory genes were downloaded from the HPA database. The diagnostic secretory RNAs of gastric cancer were screened using least absolute shrinkage and selection operator regression analysis. Univariate Cox regression analysis was used to evaluate the prognostic significance of the results. Biological functions were performed using gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses. Then, 640 cases of gastric cancer and paracancerous tissues were collected, and immunohistochemistry (IHC) was used to detect the expression of COL4A1. Results: In total, 25 upregulated differentially expressed genes (DEGs) were identified, which were secreted mainly in the blood and cell matrices. Six secretory genes (OLFM4, CEMIP, APOC1, CST1, COL4A1, and CD55) with diagnostic significance were identified, and the enrichment scores of these six genes were significantly associated with tumor stage. In addition, we found that increased COL4A1 expression might be associated with a poor prognosis in patients with gastric cancer. Based on GO and KEGG analyses, we found COL4A1-related DEGs were mainly enriched in connective tissue development, collagen fibrous tissue-related processes, extracellular structure, extracellular matrix (ECM) tissue, and related to the ECM receptor-related pathway, focal adhesion, and PI3K-Akt signaling pathway. Moreover, the results of immunohistochemical analyses showed that the COL4A1 protein level in gastric cancers was also higher than in the matched paracancerous tissues. Conclusions: In this study, we found six upregulated secretory genes, including OLFM4, CEMIP, APOC1, CST1, COL4A1, and CD55 which we hypothesized to be significant DEGs for the diagnosis of gastric cancer. Our data also suggest that COL4A1 may play an important role in the diagnosis and prognosis of gastric cancer.


Four-branched graft inversion technique for the distal anastomosis in acute aortic dissection.
Distal anastomosis bleeding is an issue during total arch replacement with the frozen elephant trunk technique. We used the 4-branched graft inversion technique for the distal anastomosis in acute aortic dissection. The aim was to evaluate the feasibility and benefits of the technique used during the frozen elephant trunk procedure for acute aortic dissection. From January 2017 to July 2019, 109 patients underwent total arch replacement for type A acute aortic dissections. Patients were divided according to the technique used for the distal anastomosis as follows: group G (n = 57; 4-branched graft inversion technique) and group C (n = 52; conventional method with Teflon felt). The postoperative variables were analysed. The hospital mortality rate was 9.2% (10/109). The mean cardiopulmonary bypass, cardiac arrest, and circulatory arrest times were 234.95 ± 71.88 min, 168.25 ± 61.33 min, and 39.19 ± 9.45 min, respectively. The circulatory arrest and cardiac arrest times were shorter in the graft inversion group than in the conventional group (36.46 ± 7.88 min vs. 42.19 ± 10.17 min, P = 0.001 and 156.21 ± 55.99 min vs. 181.44 ± 64.68 min, P = 0.031, respectively). There were 7 cases of stroke (6.4%) and 5 cases of paraplegia (4.6%). Additionally, 13 patients (11.9%) required temporary continuous renal replacement therapy. Respiratory failure occurred in 19 patients (17.4%). There were no significant differences in postoperative complications between the two groups. The 4-branched graft inversion technique provides effective and confirmed haemostasis during total aortic arch replacement using the frozen elephant trunk procedure.


A novel approach for identification of biologically active phenolic compounds in complex matrices using hybrid quadrupole-orbitrap mass spectrometer: A promising tool for testing antimicrobial activity of hops.
The phenolic compounds, secondary metabolites of hops represent a large family of compounds that could be subsequently divided into smaller groups based on the similarities between their chemical structures. The antibacterial, antifungal and antiviral properties of hops are well known, but there is a lack of information about antimicrobial activities of individual hop compounds. This study was carried out with an objective to identify compounds present in hops that have potential antibacterial activity. In the first stage of experiment, the active compounds with potential anti-microbial activity had to be extracted from hop cones. Therefore, minced hop cones were applied on solid growth medium inoculated with Staphylococcus aureus. The active substances that migrated into the medium created an inhibition zone. In the second stage of experiment, the inhibition zones were cut out from Petri dishes, active compounds were extracted from these zones and consequently analyzed using LC-HRMS. These complex assays were developed and optimized. The data were acquired by using a quadrupole-orbitrap hybrid mass spectrometer by targeted-MS2 experiment in both ionization modes. The MS method has been developed as a screening method with a subsequent fragmentation of compound of interest on the base of inclusion mass list. The unknown compounds extracted from inhibition zones have been identified either by searching against a database or their structure has been elucidated on the basis of their fragmentation spectra. On the basis of this experiment the list of active compounds with potential anti-microbial activities was enhanced.


The effect of oxytocin on potential-dependent calcium current in snail neurons, Helix pomatia.
1. The effect of external application of oxytocin on inward calcium current in dialyzed snail neurons has been investigated under clamp conditions. 2. External application of oxytocin in a dose-dependent manner (Kd 0.9 microM) inhibits inward calcium current in dialyzed neurons of the snail, Helix pomatia. 3. Inhibition of calcium current developed with the time constant of about 2 min. The degree of restoration of calcium current after oxytocin washout depends on duration of oxytocin action. 4. It has been suggested that inhibition of calcium current by oxytocin occurs in two stages, the initial one is more fast and reversible and the second one--more slow and irreversible. The participation of soluble second messengers in the inhibitory effect of oxytocin on calcium current is discussed.


Impact of reflection on the fluence rate distribution in a UV reactor with various inner walls as measured using a micro-fluorescent silica detector.
An assessment of the impact of ultraviolet (UV) reflection from inner walls is important for the accuracy of model predictions of fluence rate (FR) distribution and for the improvement of reactor efficiency. In this study, the FR distribution in an annular UV reactor with inner walls of various reflectances was measured in-situ by using a 360° response micro-fluorescent silica detector. The tests were performed in water with various transmittances ranging from 65% to 99% and with inner reactor walls composed of quartz/aluminum foil, quartz/stainless steel, or quartz/black cloth, whose reflection coefficients were determined to be 80.5%, 26.1% and 11.1%, respectively. The results demonstrate that an inner wall with a high reflection coefficient can lead to a marked increase in the weighted average FRs, thus greatly improving the reactor efficiency. Furthermore, the presently used FR distribution models could have an error of up to 35% for commonly used stainless steel walls as a result of the influence of inner-wall reflection. Finally, it was found that the uniformity of the FR distribution is strongly dependent on the diffuse reflection property of the inner wall, which could lead to a better fluence delivery distribution in the UV reactor. This work has potential application to increase the accuracy of model predictions as well as optimize the design of high-efficiency UV reactors.


Plastic changes in lumbar segments after thoracic spinal cord injuries in adult rats: an integrative view of spinal nociceptive dysfunctions.
Spinal cord injuries (SCI) cause motor, sensory and autonomic dysfunctions as well as neuropathic pain. We investigated plastic changes occurring in cord segments caudal to the lesion to assess their potential contribution to pain states after SCI. Different thoracic SCIs were performed in adult rats. Functional and algesimetry tests were performed along 3 months. Several elements of the spinal nociceptive circuitry were assessed by immunohistochemical analyses of lumbar segments. Injured animals manifested mechanical and thermal hyperalgesia. Wind-up responses and spinal reflexes were enhanced, indicating spinal hyperexcitability. We found an increase in density of nociceptive sensory afferences and in GABA inhibitory activity in dorsal horns, and increased glial reactivity. Serotoninergic descending fibers and contacts on ventral horn motoneurons were reduced. Motoneurons presented more abundant inhibitory inputs, identified by gephyrin. Not all the changes kept direct relationship to the severity of the injury. The existence of hyperalgesia despite the boost of inhibitory elements in the spinal cord confirms the dysbalance between excitatory and inhibitory mechanisms, leading to a general disinhibition. Widespread dysfunctions in remote segments after central injuries contribute to the appearance of pain, and they may be new targets for therapies aimed to modulate spinal dysfunctions after injury.


[Parendodontal surgery of the lower third molar: report of a clinical case].
The parendodontic surgery intends to the resolution of accidental or pathological condition that can't be recoverable by usual endodontic treatment. In this case, the authors present a clinical case where a apical curettage and apicectomy were done in a mandibular right third molar, to show the account of this procedure yet in the actual context.


Bifunctional Furfuryl Cations Strategy: Three-Component Synthesis of Enamidyl Triazoles.
A new multicomponent synthesis of functionalized enamidyl triazoles starting from simple and readily available starting materials is described. A simple treatment of a dichloromethane solution of an azide, amine, and 5-bromo-2-furylcarbinol with a Lewis acid provides the enamidyl triazole in good to high yield. A triple domino sequence, formal [3+2] cycloaddition/ring-opening/amidation, is involved in this new skeleton-generating reaction.


Elevated flow rate during cardiopulmonary bypass is associated with fluid accumulation.
High flow rates during cardiopulmonary bypass are assumed to increase fluid accumulation. This study aimed to determine whether two different flow rates during cardiopulmonary bypass alter the intraoperative fluid balance and extravasation rate. Sixteen pigs underwent 60 minutes of normothermic bypass, followed by 90 minutes of hypothermic bypass. A high-flow group (HF group, n = 8) had a cardiopulmonary bypass flow rate of 110 mL x kg(-1) x min(-1) and a low-flow group (LF group, n = 8) had a rate of 80 mL x kg(-1) x min(-1). Blood chemistry, hemodynamic parameters, plasma and interstitial colloid osmotic pressure, net fluid balance, plasma volume, fluid extravasation rate, and total tissue water content were measured or calculated. Results are presented as mean (standard deviation). The average net fluid balance during cardiopulmonary bypass was 1.02 (0.25) and 0.73 (0.23) mL x kg(-1) x min(-1) in the HF group and LF group, respectively (P < .05). The average fluid extravasation rate was 0.98 (0.22) and 0.77 (0.22) mL x kg(-1) x min(-1) in the HF group and the LF group (P = .07). Total water content was higher in the kidneys (P < .05) and tended to be higher in the lungs (P = .05), liver (P = .07), and brain (P = .07) of the HF group than in those of the LF group. The between-group differences in net fluid balance and fluid extravasation rate were present during the first 30 minutes of normothermic cardiopulmonary bypass. Thereafter, the values stabilized and remained similar in the two groups. Plasma volume and systemic vascular resistance differed between the groups. Cardiopulmonary bypass flow rate of 110 mL x kg(-1) x min(-1) was associated with higher positive net fluid balance and fluid extravasation rate than 80 mL x kg(-1) x min(-1). The effect was mainly observed in the initial phase of cardiopulmonary bypass.


Detection of HPV-16 genome in human oral cancers and potentially malignant lesions from India.
The presence of high risk human papilloma virus (HPV) 16 and 18 was examined in 100 oral cancer patients of Indian descent, 80 patients with potentially malignant oral lesions and corresponding clinically normal mucosa from 48 of these patients. Additionally, presence of HPV-33, -6 and -11 was also studied in 86 oral cancers, 50 potentially malignant oral lesions and 30 corresponding normal oral mucosa. All the patients with oral cancer and oral lesions, were long term tobacco-chewers, and a majority of the patients were in Advanced Stages III and IV. The DNA samples were amplified by polymerase chain reaction (PCR) using HPV L1 consensus primers. Typing of HPV was performed by Southern hybridization analysis of the PCR products using HPV-16, -18, -33, -6 and -11 type specific oligonucleotide probes. HPV-16 was detected in 15 out of 100 (15%) oral tumours, 27 out of 80 (34%) potentially malignant lesions and 15 out of 48 (31%) of the corresponding normal mucosa in the patients with oral lesions. HPV-18 was not detected in any of the oral cancers, oral lesions and normal mucosa. HPV-33 and the low-risk HPV-6 and -11 were also not detected in the oral cancers, oral lesions and corresponding normal mucosa. A significantly higher prevalence of HPV-16 was observed in oral lesions (27 out of 80, 34%) as compared to oral cancers (15 out of 100, 15%). The observed difference of 19% (95% confidence interval [CI]: 6%, 31%), between these two proportions was statistically significant at the 5% level of significance. Our data indicates that HPV-16 may play a direct role in a certain proportion of oral cancers; whereas in a subpopulation of oral cancers HPV-16 infection may be vital in the early events associated with development of potentially malignant oral lesions, and the presence of the virus not essential in the progression of the oral lesion to frank malignancy.


The first releases of transgenic mosquitoes: an argument for the sterile insect technique.
Potential applications for reducing transmission of mosquito-borne diseases by releasing genetically modified mosquitoes have been proposed, and mosquitoes are being created with such an application in mind in several laboratories. The use of the sterile insect technique (SIT) provides a safe programme in which production, release and mating competitiveness questions related to mass-reared genetically modified mosquitoes could be answered. It also provides a reversible effect that would be difficult to accomplish with gene introgression approaches. Could new technologies, including recombinant DNA techniques, have improved the success of previous mosquito releases? Criteria for an acceptable transgenic sterile mosquito are described, and the characteristics of radiation-induced sterility are compared with that of current transgenic approaches. We argue that SIT using transgenic material would provide an essentially safe and efficacious foundation for other possible approaches that are more ambitious.


Changes in the clinical features of older patients admitted from the emergency department.
Demographic changes and healthcare reforms may impact the profile of hospitalized older persons. In this study, we sought to compare the characteristics of two prospective cohorts recruited at a ten-year interval (1999, n=253-2009, n=355). They included older patients (≥75 years) admitted through the emergency department for at least 48 h in acute non-geriatric wards in the same university hospital. The exclusion criteria were patients who were admitted directly to the intensive care unit, who were dependent for all 6 Activities of Daily Living (ADL), who had recently suffered from a major stroke, or whose with a life expectancy of less than 3 months. Median age was higher in 2009 than in 1999 (83 vs. 81; p=0.020), with a higher proportion of those aged 85 years and over (p=0.026). Patients in the 2009 cohort were less likely to live in a nursing home (p=0.018), more dependent for the basic ADL (p<0.001), more independent for the instrumental ADL (p<0.001). They were more likely to have fallen in the previous year (p<0.001). They took more medications (p<0.001). Their length-of-stay was shorter (p<0.001), but they were more likely to be discharged to a rehabilitation center (p<0.001). They underwent more early re-admissions (p=0.020) and similar 3-month functional decline (p=0.614). In conclusion, within a decade, the social, functional and medical characteristics of older patients admitted to hospital have changed significantly. In view of the high consumption of in-patient services by this population, hospitals must adapt to these rapid changes.


UASB performance and electron competition between methane-producing archaea and sulfate-reducing bacteria in treating sulfate-rich wastewater containing ethanol and acetate.
To find an appropriate method for sulfate-rich wastewater containing ethanol and acetate with COD/sulfate ratio of 1, a UASB reactor was operated for more than 180 days. The influences of HRT (hydraulic retention time) and OLR (organic loading rate) on organics and sulfate removal, gas production, and electrons utilization were investigated. The sludge activity and microorganism composition were also determined. The results indicated that this system removed more than 80% of COD and 30% of sulfate with HRT above 6h and OLR below 12.3 gCOD/L d. Further HRT decrease caused volatile fatty acids accumulation and performance deterioration. Except at HRT of 2h, COD and electron flow were mostly utilized by methane-producing archaea (MPA), and methane yield remained in the range of 0.18-0.24 LCH4/gCOD. Methane was mainly generated by Methanosaeta concilii GP6 with acetate as substrate, whereas sulfate was mainly reduced by incomplete-oxidizing Desulfovibrio species with ethanol as substrate.


PnPP-19, a spider toxin peptide, induces peripheral antinociception through opioid and cannabinoid receptors and inhibition of neutral endopeptidase.
The synthetic peptide PnPP-19 has been studied as a new drug candidate to treat erectile dysfunction. However, PnTx2-6, the spider toxin from which the peptide was designed, induces hyperalgesia. Therefore, we intended to investigate the role of PnPP-19 in the nociceptive pathway. Nociceptive thresholds were measured by paw pressure test. PnPP-19 was administered intraplantarly alone or with selective cannabinoid or opioid receptor antagonists. The hydrolysis of PnPP-19 by neutral endopeptidase (NEP) (EC 220.127.116.11), an enzyme that cleaves enkephalin, was monitored by HPLC and the cleavage sites were deduced by LC-MS. Inhibition by PnPP-19 and Leu-enkephalin of NEP enzyme activity was determined spectrofluorimetrically. PnPP-19 (5, 10 and 20 μg per paw) induced peripheral antinociception in rats. Specific antagonists of μ opioid receptors (clocinnamox), δ opioid receptors (naltrindole) and CB1 receptors (AM251) partly inhibited the antinociceptive effect of PnPP-19. Inhibition of fatty acid amide hydrolase by MAFP or of anandamide uptake by VDM11 enhanced PnPP-19-induced antinociception. NEP cleaved PnPP-19 only after a long incubation, and Ki values of 35.6 ± 1.4 and 14.6 ± 0.44 μmol·L(-1) were determined for PnPP-19 and Leu-enkephalin respectively as inhibitors of NEP activity. Antinociception induced by PnPP-19 appears to involve the inhibition of NEP and activation of CB1, μ and δ opioid receptors. Our data provide a greater understanding of the antinociceptive effects of PnPP-19. This peptide could be useful as a new antinociceptive drug candidate.


Evaluation of a pulse oximeter during profound hypothermia. An assessment of the Biox 3700 during induction of hypothermia before cardiac surgery in paediatric patients.
The accuracy of the Ohmeda Biox 3700 pulse oximeter was evaluated in 10 paediatric patients, deliberately surface cooled to 25 degrees C in preparation for cardiac surgery, by comparing the arterial oxygen saturation results obtained from an Ohmeda Biox 3700 and a Radiometer OSM-2 Hemoximeter. Though there was a good correlation between the two series of results, the arterial oxygen saturation was over-estimated by the pulse oximeter compared to the OSM-2 Hemoximeter in the temperature range 36 degrees to 30 degrees C and under-estimated below 30 degrees C. These differences were greatest when the initial saturations were low.


Binding of platinum to human transferrin.
A complex of platinum and human transferrin has been formed by appropriately combining apotransferrin (metal free protein) and potassiumchloroplatinate (K2PtCl4). Atomic absorption spectroscopy indicated that both primary bind sites on the protein participated in the complex. Electron paramagnetic resonance (EPR) examination showed that the bound platinum was not paramagnetic, and thus it is highly probable that the Pt ion is in the +2 oxidation state. The results suggest a possible mechanism for physiological distribution of third-transition-series metals.


On key lesions and all that: a tribute to Paul Lohman.
This paper is a tribute to Paul Lohman at the occasion of his retirement from the position of Professor in the Medical Faculty at the Leiden University in The Netherlands and as Director of its Department of Radiation Genetics and Chemical Mutagenesis. Paul's contributions to the science of genetic toxicology are discussed in the context of more recent insights as to how mammalian cells process DNA damage, and how this may lead to cancer and, possibly, aging. Starting with his work on the characterization of UV-induced DNA repair in cultured cells from xeroderma pigmentosum patients and the development of methodology for monitoring the removal of UV-induced lesions in human cells, the concept of the key lesion is introduced. Among the myriad of DNA lesions that can be induced in DNA as a consequence of exposure to a range of natural or synthetic mutagens, key lesions are the ones responsible for subsequent adverse effects, for example, because they give rise to mutation. The development of methods using immunofluorescence microscopy to detect and identify such key lesions and quantitate them at the single cell level, is one of the highlights of Paul's career. Based on the perceived need to evaluate mutational end points in vivo in relation to specific lesions identified by his immunofluorescence methods, Paul subsequently made crucial contributions to the development of the first transgenic mouse model to measure mutations in chromosomally integrated reporter genes. In parallel to his experimental work, Paul greatly contributed to genetic toxicology at the theoretical level by his work on the development and evaluation of methods for assessment or prediction of risks of exposure to environmental mutagens. Finally, Paul has served the discipline of genetic toxicology in a more administrative role in various ways, both locally as one of the founders of the Medical Genetics Center South-West Netherlands and internationally by playing a prominent role in organizations such as ICPEMC. Here, his numerous contributions to the journal Mutation Research, both as author on many papers and as Executive Managing Editor should not go unmentioned.


Healing through the lens of immunothrombosis: Biology-inspired, evolution-tailored, and human-engineered biomimetic therapies.
Evolution, from invertebrates to mammals, has yielded and shaped immunoclotting as a defense and repair response against trauma and infection. This mosaic of immediate and local wound-sealing and pathogen-killing mechanisms results in survival, restoration of homeostasis, and tissue repair. In mammals, immunoclotting has been complemented with the neuroendocrine system, platelets, and contact system among other embellishments, adding layers of complexity through interconnecting blood-born proteolytic cascades, blood cells, and the neuroendocrine system. In doing so, immunothrombosis endows humans with survival advantages, but entails vulnerabilities in the current unprecedented and increasingly challenging environment. Immunothrombosis and tissue repair appear to go hand in hand with common mechanisms mediating both processes, a fact that is underlined by recent advances that are deciphering the mechanisms of the repair process and of the biochemical pathways that underpins coagulation, hemostasis and thrombosis. This review is intended to frame both the universal aspects of tissue repair and the therapeutic use of autologous fibrin matrix as a biology-as-a-drug approach in the context of the evolutionary changes in coagulation and hemostasis. In addition, we will try to shed some light on the molecular mechanisms underlying the use of the autologous fibrin matrix as a biology-inspired, evolution-tailored, and human-engineered biomimetic therapy.


Dietary management for older subjects with obesity.
In recent years, obesity has been recognized as a form of malnutrition in older adults and a continuing risk factor for serious health problems. Weight reduction in older adults is not as reliable a recommendation as it is for younger adults; a decreased body mass index (BMI) seems to be associated with a higher incidence of stroke, and a normal or slightly elevated BMI has been linked to greater reserve capacity. Weight loss in older adults requires strategies that consider health status, functional ability, and rational targets. Strategies may include behavior modification, dietary alterations, exercise or physical activity, and reasonable goals that do not put the individual at nutritional risk. Studies that examine different approaches to weight reduction rarely include older subjects, so it is difficult to make judgments about various interventions (surgery, exercise, drugs, or diet) and their efficacy in this population. Fad diets may be lacking in essential nutrients and may prove to be risky for elderly people. Weight loss programs for older adults should focus on maintaining adequate intake of essential nutrients while reducing calories by controlling dietary fat intake.


KHK-A promotes the proliferation of oesophageal squamous cell carcinoma through the up-regulation of PRPS1.
The metabolism of dietary fructose by ketohexokinase (KHK) is an important step in glucose metabolism in various tumour types. However, the expression, function and underlying mechanisms of KHK in oesophageal squamous cell carcinoma (ESCC) remain largely unclear. The objective of this study was to investigate the effects of KHK-A, a peripheral isoform of KHK, on the proliferation of ESCC cell lines. The function and mechanism of KHK-A in ESCC cells were investigated by constructing stable KHK-A-knockdown and -overexpressing ESCC cell lines (KYSE410 and KYSE150, respectively). The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide, flow cytometry and colony formation assays were used to analyse the effects of KHK-A on cell proliferation, cell cycle and colony formation, respectively. KHK-A and phosphoribosyl pyrophosphate synthetase isoform 1 (PRPS1) mRNA and protein expressions in several ESCC cell lines were determined using routine reverse transcription-polymerase chain reaction and immunoblotting, respectively. KHK and PRPS1 expressions in ESCC tumour tissues and corresponding adjacent non-tumour tissues were evaluated according to the gene expression omnibus (GEO) database (GSE20347). In vitro experiments showed that KHK-A significantly promoted cell proliferation by modulating the G1/S phase transition in the cell cycle, which was probably regulated by PRPS1 expression. GEO database-based analysis showed that KHK levels were significantly higher in the ESCC tissues than in the corresponding adjacent non-tumour tissues. Pearson's correlation coefficient analysis showed that KHK expression in ESCC cell lines and tissues was significantly positively associated with the up-regulation of PRPS1, suggesting that KHK-A levels regulate PRPS1 expression in ESCC. KHK-A may serve as a driving gene in ESCC for the activation of PRPS1, resulting in the up-regulation of PRPS1. This could lead to enhanced nucleic acid synthesis for tumourigenesis. Our study showed that KHK-A is a potential target for ESCC diagnosis and therapy.


"Formal" ruthenium-catalyzed [4+2+2] cycloaddition of 1,6-diynes to 1,3-dienes: formation of cyclooctatrienes vs vinylcyclohexadienes.
[reaction: see text] A new "formal" Ru-catalyzed [4+2+2] cycloaddition of 1,6-diynes to 1,3-dienes giving conjugated 1,3,5-cyclooctatrienes and vinylcyclohexadienes is described. This formal cycloaddition is really a tandem process, the Ru(II)-catalyzed formation of (Z)-tetraenes or vinyl-(Z)-trienes followed by a pure thermal conrotatory 8 pi- or disrotatory 6 pi-electrocyclization. The proposed mechanism allows the differences in product ratio to be explained in terms of steric and stereochemical considerations.


Nomogram for individualizing supplementary iron doses during erythropoietin therapy in haemodialysis patients.
An adequate iron supplement is crucial not only for prompt erythropoiesis but also for the restoration of tissue iron reserves in haemodialysis patients receiving recombinant human erythropoietin (r-HuEPO). An attempt was made to establish a comprehensive nomogram that allows individualization of intravenous (i.v.) iron doses according to patients' body weights, the initial status of tissue iron reserves and desired increases in haemoglobin levels. Clinical and laboratory data retrieved from 95 haemodialysis patients who received r-HuEPO with or without iron supplements for at least 12 weeks were used to construct the nomogram. It was assumed that the administered iron was either incorporated into newly synthesized haemoglobin and tissue iron reserves or eliminated from the body at a constant rate. Tissue iron reserves of the patients were estimated by serum ferritin levels using van Wyck's equation (Kidney Int., 1989, 35, 712). The rate of iron loss in the patients was estimated by the data obtained from 15 of the above patients who exhibited stable haemoglobin levels but decreases in serum ferritin levels with no iron supplements. The validity of the equation was ascertained by comparing the measured serum ferritin levels at the end of r-HuEPO therapy and those predicted by the nomogram. The proposed nomogram was then validated prospectively in 24 haemodialysis patients to determine whether the nomogram-recommended iron doses would increase both haemoglobin and serum ferritin levels within 12 weeks. The mean (+/-SD) iron loss of haemodialysis patients was calculated to be 10.5 +/- 7.4 mg/week. There was a significant correlation (r=0.77, P < 0.001) between the measured serum ferritin levels, an index of tissue iron reserves, at the end of r-HuEPO therapy and those predicted by the equations used for formulating the nomogram. The prospective study indicated that the nomogram-recommended supplementary iron doses attained haemoglobin and serum ferritin levels of > 95 g/L and >100 microg/L in 79 and 50% of the patients, respectively, within 12 weeks. The present nomogram may be useful for individualizing supplementary i.v. iron doses for haemodialysis patients undergoing r-HuEPO therapy.


Plasmodium berghei malaria: effect of acute phase serum on immunity generated in rats by infection and by vaccination.
Acute phase serum (APS) given at the time of challenge with Plasmodium berghei inhibited the generation to immunity to the infecting plasmodia. Administered with a single dose of vaccine, it inhibited induction of immunity by the vaccine. Three weekly doses, the last given two weeks before infection, induced immunity. Administration of vaccine simultaneously with infection neither aggravated nor ameliorated the infection. These results indicate that the effect of administration of APS on immunity generated by immunization or infection is dose- and time-dependent. The depression of immunity induced by this serum is thus temporary, the host finally overcoming the depression and responding to the plasmodial antigen in the serum. The interaction of vaccine and infection observed indicates that the introduction of vaccine is not detrimental to the individual incubating infection; rather, the vaccine is rendered useless, the reducing the aggregate benefit of the immunization to the group.


Heparin affects matrix metalloproteinases and tissue inhibitors of metalloproteinases circulating in peripheral blood.
Blood sampling/handling alters matrix metalloproteinases (MMP) and tissue inhibitors of metalloproteinases (TIMP) expression. The aim of this study is to evaluate the effects of high molecular weight heparin on MMP and TIMP expression in blood. We analyzed by gelatin zymography and ELISA assays the effects of different heparin salts, dose- and time-dependence of MMP and TIMP concentrations in plasma and sera collected with and without clot-accelerator in plastic tubes from 50 healthy donors. The levels and zymography of MMP-2 did not show significant changes among all samples, and during time- and dose-dependent heparin treatments. MMP-9 and TIMP-2 expression were strongly affected by heparin, with significant increase of their content and gelatinolytic activity both in time- and in dose-dependent fashion. Addition of heparin allowed also the displacement of MMP-2 prodomain, favouring zymogen activation. Heparin has direct and indirect effects, altering MMP/TIMP complexes circulating in blood, and increasing the release of TIMP-2. To avoid misinterpretations due to MMP/TIMP complex alteration and MMP prodomain displacement, heparin should be cautiously used in blood collection procedures.


Digitisation of the total burn surface area.
The assessment of surface area of the body affected by a burn (TBSA) has long been estimated with manual charts. Initial assessment of burned patients is made frequently by clinicians with limited experience producing significant errors. Paper copies of burn charts are unwieldy, subject to loss and tend towards overestimation. Thus, a simple method of calculation, recording and transmission via email or telemedicine may produce benefits in both initial treatment and data recording. Although computer-based systems have been reported previously none have entered routine clinical practice in the UK. We devised a PC-based program, "Burn Calculator", whereby digital transcription of the burn allows automatic area calculation allowing not only a rapid, accurate figure for determination of fluid resuscitation, but also the potential for rapid electronic transmission. It also calculates fluid requirements to minimise errors during resuscitation. This initial pilot study compared figures from 50 paper charts with those from Burn Calculator to determine its accuracy and reproducibility. Previously reported variations in TBSA estimation were confirmed, as was the tendency towards TBSA underestimation resulting from transcription of a three-dimensional clinical situation to a two-dimensional representation. Burn Calculator showed high correlation (r=0.9850; p<0.0001) and reproducibility (R=0.9957) that would simplify assessment and referral plus facilitate data collection, interpretation and research.


Atrioventricular node reentrant tachycardia in patients with a prolonged AH interval during sinus rhythm: clinical features, electrophysiologic characteristics and results of radiofrequency ablation.
Among a consecutive series of 600 patients who underwent radiofrequency catheter ablation for AV node reentrant tachycardia, 14 patients (age 29-76 years) had a prolonged AH interval during sinus rhythm (172 +/- 18 ms, range 140 to 200). Seven of them had unsuccessful ablation during the previous ablation sessions. Eight patients with anterograde dual AV node pathway physiology received anterograde slow pathway ablation, and the other 6 patients without dual-pathway physiology received retrograde fast pathway ablation. All patients had successful elimination of AV nodal reentrant tachycardia after a mean of 4 +/- 4 radiofrequency applications, power level 36 +/- 6 watts and a pulse duration of 42 +/- 4 seconds. The postablation AH interval remained unchanged. During a follow-up period of 25 +/- 13 months, one patient who received slow pathway ablation developed 2:1 AV block with syncope. As compared with the other 586 patients without a prolonged AH interval, these 14 patients had significantly poorer anterograde AV nodal function and lower incidence of anterograde dual AV node physiology (P < 0.01). We concluded that slow pathway ablation in patients with dual pathway physiology, and retrograde fast pathway ablation in patients without dual pathway physiology were effective and safe in patients with a prolonged AH interval. However, delayed onset of symptomatic AV block is possible and careful follow-up is necessary.


Thermosensitive molecularly imprinted poly(1-vinyl-2-pyrrolidone/methyl methacrylate/N-vinylcaprolactam) for selective extraction of imatinib mesylate in human biological fluid.
The efficiency of a molecularly imprinted polymer as a selective packing material for the solid-phase extraction of imatinib mesylate sorption was investigated. The molecularly imprinted polymer was prepared using N,N'-methylenebisacrylamide as a cross-linker agent, N-vinylcaprolactam as a thermo-sensitive monomer, 1-vinyl-2-pyrrolidone and methyl methacrylate as functional monomers, azobisisobutyronitrile as an initiator and imatinib mesylate as a template. The drug-imprinted polymer was identified by Fourier transform infrared spectroscopy, thermogravimetric analysis, elemental analysis, and scanning electron microscopy. It was found that this polymer can be used for determination of trace levels of imatinib mesylate with a recovery percentage that could reach over 90%. Furthermore, the synthesized molecularly imprinted polymer indicated higher selectivity towards imatinib mesylate than other compounds. From isotherm study, the equilibrium adsorption data of imatinib mesylate by imprinted polymer were analyzed by Langmuir, Freundlich, and Temkin isotherm models. The developed method was used for determination of imatinib mesylate in human fluid samples by high performance liquid chromatography with excellent results.


A macromolecular inhibitor of in vitro calcification of tendon matrix.
Bovine and human tendon tissue do not induce calcification in vitro. However, extraction of those tissues with 3% Na2HPO4 converts them to calcifiable matrices. The supernatant fraction derived from the extraction contains a nondialyzable, perchloric acid soluble component that inhibits calcification of the extracted matrix. This inhibitory substance is characterized by a molecular weight in the range of 85,000-100,000. Exposure to pronase or hyaluronidase did not alter the inhibitory potency but did render the inhibitor dialyzable. Commercial sources of hyaluronic acid, chondrotitin-6-sulfate, chrondroitin-4-sulfate, dermatan sulfate, heparin and lysozyme did not inhibit calcification of the extracted matrix. Phosvitin, a phosphoglycoprotein is a potent inhibitor. Although phosvitin and the tendon extract also inhibit calcification of previously calcified matrix, they have no detectable effect on the rate of decalcification. We conclude that the mechanism of inhibition is characterized by a degree of specificity and that phosvitin and a macromolecular component of tendon tissue blocks conversion of an intermediate matrix-bound CaP complex to crystalline apatite. It seems reasonable that the tendon inhibitor could function in situ and possibly in vivo to control calcification of tendon tissue.


Evaluating Nighttime Observational Measures of Neighborhood Disorder: Validity of the Nighttime NIfETy Assessment.
While there are a growing number of observational instruments to assess the built and social dimensions of the neighborhood environment, there are few reliable and validated instruments; there are no instruments that assess the neighborhood environment during nighttime hours, a potential peak period of health and safety risk. The purpose of this investigation is to establish the metric properties of Neighborhood Inventory for Environmental Typology (NIfETy) Instrument nighttime ratings. Reliability of the scale was measured by internal consistency reliability and test re-test correlation. Validity was evaluated through correlation with the daytime NIfETy rating and regression models with local violent crime data. The nighttime items had good internal consistency (α = .81-82) for the total scale and acceptable internal consistency for a seven-item nighttime disorder scale (α = .66-.71). Future investigations will examine the nighttime NIfETy and its association with specific risk behaviors to evaluate changes in neighborhood environment.


The first 90 days following release from jail: findings from the Recovery Management Checkups for Women Offenders (RMCWO) experiment.
(1) To examine the impact of monthly Recovery Management Checkups (RMC) vs. control in the first 90 days post-release from jail on receipt of community-based substance abuse treatment, and (2) To explore the impact of RMC, treatment, and abstinence on HIV risk behaviors and recidivism. Of the 480 women randomized, 100% completed the intake and release interviews, and over 90% completed the 30-, 60-, and 90-day post-release interviews. Of the 915 times women assigned to RMC were interviewed (at release, 30, 69 and 90 days post release), 885 (97%) times they attended linkage meetings, 429 (47%) times they were identified as in need of substance abuse treatment, 271 (30%) times they agreed to go to treatment, 149 (16%) times they showed to the treatment intake, and 48 (5%) times they stayed in treatment at least two weeks. During the 90 days following release from jail, women in the RMC condition (vs. control) were significantly more likely to return to treatment sooner and to participate in substance abuse treatment. Women who received any treatment were significantly more likely than those who did not to be abstinent from any alcohol or other drugs. Those who were abstinent were significantly more likely to avoid HIV risk behaviors and recidivism. These results demonstrate the feasibility of conducting monthly Recovery Management Checkups with women offenders post-release and provide support for the effectiveness of using RMC to successfully link women offenders to treatment.


Discovery of cell-permeable non-peptide inhibitors of beta-secretase by high-throughput docking and continuum electrostatics calculations.
A fragment-based docking procedure followed by substructure search were used to identify active-site beta-secretase inhibitors from a composite set of about 300 000 and a library of nearly 180 000 small molecules, respectively. EC(50) values less than 10 microM were measured in at least one of two different mammalian cell-based assays for 12 of the 72 purchased compounds. In particular, the phenylureathiadiazole 2 and the diphenylurea derivative 3 are promising lead compounds for beta-secretase inhibition.


HIV, HEV and cirrhosis: evidence of a possible link from eastern Spain.
The aim of the study was to assess the seroprevalence of hepatitis E virus (HEV) infection in an HIV-infected population, as determined by HEV immunoglobulin G (IgG) antibodies (anti-HEV). The design of the study was cross-sectional. Serum anti-HEV IgG was determined by enzyme immunoassay in 238 HIV-infected patients consecutively attending our out-patient clinic between April and May 2011. In HEV-seropositive patients, HEV RNA was analysed by nested reverse transcriptase-polymerase chain reaction (RT-PCR). Associations between anti-HEV and liver cirrhosis, route of HIV infection, hepatitis B virus (HBV) and hepatitis C virus (HCV) serological markers, age, sex and alanine aminotransferase (ALT) levels were examined by univariate and multivariate analysis. One hundred and forty patients (59%) had chronic liver disease (99% were HBV- and/or HCV-coinfected). Liver cirrhosis was detected in 44 individuals (19%). Two hundred and twelve patients (89%) were on antiretroviral treatment; the median CD4 T-cell count was 483 cells/μL [interquartile range (IQR) 313-662 cells/μL] and the HIV viral load was <25 HIV-1 RNA copies/mL. Overall, 22 patients (9%) were anti-HEV positive. Liver cirrhosis was the only factor independently associated with the presence of anti-HEV, which was documented in 23% of patients with cirrhosis and 6% of patients without cirrhosis (P=0.002; odds ratio 5.77). HEV RNA was detected in three seropositive patients (14%), two of whom had liver cirrhosis. Our findings show a high prevalence of anti-HEV in HIV-infected patients, strongly associated with liver cirrhosis. Chronic HEV infection was detected in a significant number of HEV-seropositive patients. Further research is needed to ascertain whether cirrhosis is a predisposing factor for HEV infection and to assess the role of chronic HEV infection in the pathogeneses of cirrhosis in this population.


Dimerization of propargyl and homopropargyl 6-azido-6-deoxy-glycosides upon 1,3-dipolar cycloaddition.
Copper-catalyzed, thermal or microwave promoted 1,3-dipolar cycloaddition (Click Reaction) of 2-propynyl and 3-butynyl 2,3,4-tri-O-acetyl-6-azido-6-deoxy-glycopyranosides in the D-gluco, D-galacto and D-manno series afford the corresponding dimeric cycloaddition products.


Smoking and physical inactivity increase cancer prevalence in BRCA-1 and BRCA-2 mutation carriers: results from a retrospective observational analysis.
The aim of this analysis in a pilot study population was to investigate whether we can verify seemingly harmful lifestyle factors such as nicotine and alcohol indulgence, obesity, and physical inactivity, as well as a low socioeconomic status for increased cancer prevalence in a cohort of BRCA 1 and 2 mutation carriers. The analysis data are derived from 68 participants of the lifestyle intervention study LIBRE-1, a randomized, prospective trial that aimed to test the feasibility of a lifestyle modification in BRCA 1 and 2 mutation carriers. At study entry, factors such as medical history, lifestyle behavior, and socioeconomic status were retrospectively documented by interview and the current BMI was determined by clinical examination. The baseline measurements were compared within the cohort, and presented alongside reference values for the German population. Study participants indicating a higher physical activity during their adolescence showed a significantly lower cancer prevalence (p = 0.019). A significant difference in cancer occurrence was observed in those who smoked prior to the disease, and those who did not smoke (p < 0.001). Diseased mutation carriers tended to have a lower BMI compared to non-diseased mutation carriers (p = 0.079), whereas non-diseased revealed a significantly higher physical activity level than diseased mutation carriers (p = 0.046). The present data in this small cohort of 68 mutation carriers suggest that smoking and low physical activity during adolescence are risk factors for developing breast cancer in women with BRCA1 or BRCA2 mutation. Further data of the ongoing LIBRE 2 study are necessary to confirm these findings in a larger cohort of 600 mutation carriers.


Intrathecal rimantadine induces motor, proprioceptive, and nociceptive blockades in rats.
The purpose of the experiment was to evaluate the local anesthetic effect of rimantadine in spinal anesthesia. Rimantadine in a dose-dependent fashion was constructed after intrathecally injecting the rats with four different doses. The potency and duration of rimantadine were compared with that of the local anesthetic lidocaine at producing spinal motor, nociceptive, and proprioceptive blockades. We demonstrated that intrathecal rimantadine dose-dependently produced spinal motor, nociceptive, and proprioceptive blockades. On the 50% effective dose (ED50) basis, the ranks of potencies at inducing spinal motor, nociceptive, and proprioceptive blockades was lidocaine>rimantadine (P<0.01). Rimantadine exhibited more nociceptive block (ED50) than motor block (P<0.05). At equi-anesthetic doses (ED25, ED50, and ED75), the spinal block duration produced by rimantadine was longer than that produced by lidocaine (P<0.01). Furthermore, rimantadine (26.52μmol/kg) prolonged the nociceptive nerve block more than the motor block (P<0.001). Our preclinical data showed that rimantadine, with a more sensory-selective action over motor block, was less potent than lidocaine. Rimantadine produced longer duration in spinal anesthesia when compared with lidocaine.


Recovery of silver from silver(I)-containing solutions in bioelectrochemical reactors.
A novel approach was tested for metallic silver recovery and power generation by using cathodic reduction in bioelectrochemical systems (BESs). In dual-chamber BESs (130 mL volume) with acetate as electron donor on anode, both Ag(+) ions and Ag(I) thiosulfate complex in catholyte were reduced on cathode. The reduction rate of Ag(+) was more rapid than the Ag(I) complex as expected by energetic analysis. X-ray diffraction (XRD) analysis indicated that electrodeposits on cathodes from both catholyte were metallic silver with >91% purity. The feasibility of metallic silver recovery with the BESs was confirmed using simulated photographic wastewater and up to 95% of Ag(I) removal was achieved.


Acetaminophen usage patterns and concentrations of glutathione and gamma-glutamyl transferase in alcoholic subjects.
To determine if subjects with chronic alcoholism are predisposed to acetaminophen-induced hepatotoxicity, and to determine the contributing factors. Prospective cohort study. Community-based crisis intervention center. One hundred eighty-eight subjects who answered "yes" to at least one of the four questions on the CAGE (Cut down-Annoyed-Guilty-Eye-opener) questionnaire for identifying alcoholism, and 10 healthy volunteers (controls). A history, physical examination, urine toxicologic analysis, ethanol and drug therapy history, and venous blood samples were collected on all subjects. Venous blood was analyzed for a liver profile, prothrombin time, and total and oxidized glutathione concentrations. A significantly higher proportion of daily drinkers were regular daily users (29.2% [45/154] vs 11.8% [4/34], p=0.0497) as well as abusers (35.7% [55/154] vs 14.7% [5/34], p=0.0237) of acetaminophen compared with non daily drinkers. Alcoholic subjects with elevated gamma-glutamyl transferase (GGT >or= 51 U/L) levels had significantly lower median plasma glutathione concentrations (2.33 micromol/L, 95% confidence interval [CI] 1.74-2.69 micromol/L) compared with those of alcoholic subjects with normal GGT concentrations (5.97 micromol/L, 95% CI 4.39-7.03 micromol/L, p<0.0001) and healthy volunteers (6.59 micromol/L, 95% CI 4.79-9.65 micromol/L, p=0.0002). A significant inverse correlation was also noted between the GGT concentration and the plasma total glutathione concentration (r = -0.62, p<0.0001). None of the 188 subjects met all preset criteria for hepatotoxicity. Daily drinkers were more than twice as likely as non daily drinkers to be regular daily acetaminophen users and abusers. Alcoholic subjects with elevated GGT concentrations had significantly lower plasma total glutathione concentrations, and plasma total glutathione concentrations inversely correlated with GGT concentrations. Elevated GGT concentrations may be a clinical marker of depleted glutathione in alcoholic subjects. Acetaminophen-induced hepatotoxicity appears to be uncommon in alcoholic subjects, despite the 31.9% (60/188 patients) who took doses that are potentially hepatotoxic.


An atlas of transcriptionally defined cell populations in the rat ventral tegmental area.
The ventral tegmental area (VTA) is a complex brain region that is essential for reward function and frequently implicated in neuropsychiatric disease. While decades of research on VTA function have focused on dopamine neurons, recent evidence has identified critical roles for GABAergic and glutamatergic neurons in reward processes. Additionally, although subsets of VTA neurons express genes involved in the synthesis and transport of multiple neurotransmitters, characterization of these combinatorial populations has largely relied on low-throughput methods. To comprehensively define the molecular architecture of the VTA, we performed single-nucleus RNA sequencing on 21,600 cells from the rat VTA. Analysis of neuronal subclusters identifies selective markers for dopamine and combinatorial neurons, reveals expression profiles for receptors targeted by drugs of abuse, and demonstrates population-specific enrichment of gene sets linked to brain disorders. These results highlight the heterogeneity of the VTA and provide a resource for further exploration of VTA gene expression.


The effects of N-methylformamide on artificial and spontaneous metastases from a murine hepatocarcinoma.
The effects of the differentiation-inducing polar solvent N-methylformamide (NMF) on artificially induced and spontaneous metastases from a murine hepatocarcinoma (HCA-1) in C3Hf/Kam mice were investigated. Exposure of HCA-1 cells in vitro for 6 days to 1.0% or 1.25% NMF resulted in an increase in the number of lung nodules formed in mice when these cells were injected into their tail veins. This in vitro NMF exposure increased cell volume and induced only a slight amount of cytotoxicity. Administration of NMF to mice 1 day before i.v. tumour cell inoculation resulted in a dose-dependent increase in the number of lung nodules formed, beginning at an NMF dose of 600 mg kg-1. NMF caused a similar magnitude of metastasis enhancement in immunosuppressed mice. However, when the maximum dose tested (1,800 mg kg-1) was administered as 6 daily fractions of 300 mg kg-1 each, no increase in artificial metastases was detected. Administration of NMF to mice one day after i.v. tumour cell injection resulted in a dose-dependent decrease in the number of lung nodules. In mice bearing 5-6 mm HCA-1 leg tumours, treatment with 6 daily fractions of NMF (300 mg kg-1 each) significantly reduced the number of spontaneous pulmonary metastases, yet had very little effect on the growth of the primary tumour. These data suggest that, in a clinically relevant treatment setting, NMF can reduce metastasis formation.


The effect of EDTA, NTA and picolinic acid on Th(IV) mobility in a ternary system with natural sand.
Organic complexing agents, such as ethylenediaminetetraacetic acid (EDTA), nitrilotriacetic acid (NTA) and picolinic acid, have been widely used at nuclear sites and are therefore found as common co-contaminants in radioactive contaminated land. This study has explored the mechanisms by which these three complexing agents affect the sorption of Th(IV) to pure silica and a natural sand. EDTA, NTA and, to a lesser extent, picolinic acid decreased the sorption of Th to silica, demonstrating the formation and solubility of Th complexes. However, Th sorption to sand was kinetically controlled and complexation enhanced the rate of Th sorption. EDTA and NTA did not sorb significantly to the sand, and metal desorption indicated that the mechanism involved exchange with sand-associated metals. At equilibrium, however, Th sorption was not affected by the presence of the ligands, and modelling suggested that the interaction between Th and the surface binding sites controlled Th sorption thermodynamically.


A Longitudinal Examination of Work-Life Balance in the Collegiate Setting.
The literature regarding work-life balance (WLB) is plentiful, particularly in the National Collegiate Athletic Association Division I collegiate setting due to its demanding nature as defined by work hours and travel. Despite the comprehensive understanding of factors contributing to WLB for athletic trainers (ATs) in this setting, the effect of time of year has yet to be investigated. To determine the influence sport season timing (ie, preseason, in-season, off-season) can have on perceptions and experiences of WLB for ATs. Qualitative, case-study design. Division I collegiate practice setting. Six ATs employed in the Division I college setting (3 women, 3 men) volunteered and completed our study over 1 year. The average age of participants was 31.0 ± 3 years, and they had 9.0 ± 3 years of experience. We conducted semistructured one-on-one phone interviews with each participant at 4 points during the year (August, November, February, June). All interviews were transcribed verbatim. Data were analyzed inductively, and we used peer review, multiple-analyst triangulation, and member checks for credibility. Time of year (eg, competitive in-season), organizational demands (eg, hours and travel), and life stage (eg, roles assumed) were factors that influenced WLB for ATs in the collegiate setting. Having a positive and adaptable mind-set; having coworker, supervisor, and personal support; and engaging in personal or family time were factors that counterbalanced the inhibitors. Our results indicate that a career as an AT in the collegiate setting can create WLB concerns; these concerns, however, seem to peak during the competitive in-season for the AT's primary coverage team. Additionally, conflict that arises for ATs seems to be stimulated by time, as work roles are often less accommodating due to the athletic atmosphere.


[Hygiene in ophthalmological practices : Results from ophthalmological practices in Frankfurt am Main in 2012-2015].
Hygiene regulations must be taken into consideration for all diagnostic and therapeutic measures in order to avoid transfer of pathogens and infections. The Infection Protection Act assigns the responsibility for prevention of infections to managers of medical institutions, i.e. the owners of a practice and public health authorities are obligated to counsel and monitor medical institutions with respect to hygiene and prevention of infections. This article presents the results of the surveillance of all ophthalmological practices in Frankfurt am Main in 2012-2015, which are assessed and discussed against the background of the valid recommendations of the Committee for Hospital Hygiene (KRINKO). Based on a checklist, all 35 ophthalmological practices in Frankfurt am Main received counseling and were monitored by an official from the Public Health Office. The basic principles of hygiene organization as well as the prerequisites for appropriate hand hygiene and surface disinfection were present in the majority of practices. In 11 practices (31.4 %) disposable medical products were exclusively utilized but medical products were prepared in-house in 24 practices (68.6 %). Contact pads for tonometry were prepared in-house in 13 (37.1 %) practices of which 6 were deficient so that improvements had to be implemented. In 6 out of the 7 practices where even critical medical products were prepared, this preparation was prohibited due to significant errors and the practices had to switch to disposable materials. In total, the practices could be certified as having a good general level of hygiene; however, in some cases major errors were found in the preparation of medical products, which had to be immediately corrected.


Measuring demand and supply of community care services for older people healthy ageing in rural Zhejiang Province.
A consequence of demographic trends and economic prosperous is the increasing diversity in needs for care services. However, the traditional family support for older people has been largely supplanted by the wider provision of community care services. This study aims to investigate the current status of demand and supply in community care services across different villages of Zhejiang province and assess service effectiveness for healthy ageing. A questionnaire survey was carried out towards 207 rural villages across 9 cities in Zhejiang province. One hundred eighty-six valid responses were received, representing a response rate of around 89.9%. Descriptive statistics were employed to identify older villagers' care needs and available community services. Comparative analysis examined the balance between the demand and supply of community care services. Correlation analysis were applied to determine key factors that impacted the supply of social services in rural communities. The research found that rural older residents normally lived with their children or spouse with limited literacy and income. On average the categories of community care services is substantially small in comparison with the increasingly diverse demands of older people in rural areas. There was an obvious mismatch between service demand and service supply in rural communities, which often caused the waste of public resources. Moving forward, the uppermost priority is given to infrastructure construction service and daily life service, while little attention is paid to mental health service and specialized nursing service. There needs to be an improvement in the socio-economic capacity of rural communities and in the diversity of social care services. Policies and strategies are also needed to encourage private sectors' involvement in providing care services for rural older people. Local government should have a clear vision of the potential demands for community care services, practical guidelines will be useful in guaranteeing better service quality.


Peripheral suppression of first pain and central summation of second pain evoked by noxious heat pulses.
Psychophysical experiments were carried out on 6 huma subjects to determine how first and second pain are influenced by peripheral receptor mechanisms and by central nervous system inhibitory and facilitatory mechanisms. For these experiments, brief natural painful stimuli delivered to the hand were a train of 4-8 constant waveform heat pulses generated by a contact thermode (peak temp. = 51-5% C). The magnitude of first and second pain sensations was estimated using cross-modality matching procedures and reaction times were determined. The latter confirmed the relationship between first and second pain and impulse conduction in Adelta and C noxious heat afferents, respectively. The intensity of first pain decreased with each successive heat pulse when the interpulse interval was 80 sec or less. This decrease was most likely the result of heat induced suppression of Adelta heat nociceptors since it did not occur if the probe location changed between successive heat pulses. In contrast, second pain increased in intensity with each successive heat pulse if the interval was 3 sec or less. This summation was most likely due to central nervous system summation mechanisms since it also occurred after blockage of first pain by ulnar nerve compression and when the location of the thermode changed between heat pulses. These observations and their interpretations are supported by our recording of responses of singlt Adelta heat nociceptive afferents, C polymodal nociceptive afferents, and "warm" afferents of rhsus monkeys to similar trains of noxious heat pulses. Their responses to these heat pulses show a progressive suppression. Furthermore, previous studies have shown that wide dynamic range dorsal horn neurons show summated responses to repeated volleys in C fibers (greater than 1/3 sec). These spinal cord summation mechanisms could account for the summation of second pain.


A Comprehensive Review of Endoscopic Management of Sleeve Gastrectomy Leaks.
Bariatric surgery leaks result in significant morbidity and mortality. Experts report variable therapeutic approaches, without uniform guidelines or consensus. To review the pathogenesis, risk factors, prevention, and treatment of gastric sleeve leaks, with a focus on endoscopic approaches. In addition, the efficacy and success rates of different treatment modalities are assessed. A comprehensive review was conducted using a thorough literature search of 5 online electronic databases (PubMed, PubMed Central, Cochrane, EMBASE, and Web of Science) from the time of their inception through March 2020. Studies evaluating gastric sleeve leaks were included. MeSH terms related to "endoscopic," "leak," "sleeve," "gastrectomy," "anastomotic," and "bariatric" were applied to a highly sensitive search strategy. The main outcomes were epidemiology, pathophysiology, diagnosis, treatment, and outcomes. Literature search yielded 2418 studies of which 438 were incorporated into the review. Shock and peritonitis necessitate early surgical intervention for leaks. Endoscopic therapies in acute and early leaks involve modalities with a focus on one of: (i) defect closure, (ii) wall diversion, or (iii) wall exclusion. Surgical revision is required if endoscopic therapies fail to control leaks after 6 months. Chronic leaks require one or more endoscopic, radiologic, or surgical approaches for fluid collection drainage to facilitate adequate healing. Success rates depend on provider and center expertise. Endoscopic management of leaks post sleeve gastrectomy is a minimally invasive and effective alternative to surgery. Their effect may vary based on clinical presentation, timing or leak morphology, and should be tailored to the appropriate endoscopic modality of treatment.


Avoidance of retrograde catheterization by Doppler echocardiographic diagnosis of Björk-Shiley aortic valve dysfunction.
A 74-year-old man with a history of prior Björk-Shiley aortic valve replacement was admitted with chest pain and dyspnea. Results of physical examination and cinefluoroscopy suggested a dysfunction of the prosthetic valve; imaging echocardiography was not helpful. Nonimaging continuous wave Doppler examination confirmed severe prosthetic aortic valve dysfunction, necessitating emergent replacement, and potentially hazardous retrograde catheterization of the prosthesis was avoided.


Hearing impairment and vowel production. A comparison between normally hearing, hearing-aided and cochlear implanted Dutch children.
This study investigated the acoustic characteristics of the Belgian Standard Dutch vowels in children with hearing impairment and in children with normal hearing. In a balanced experimental design, the 12 vowels of Belgian Standard Dutch were recorded in three groups of children: a group of children with normal hearing, a group with a conventional hearing aid and a group with a cochlear implant. The formants, the surface area of the vowel space and the acoustic differentiation between the vowels were determined. The analyses revealed that many of the vowels in hearing-impaired children showed a reduction of the formant values. This reduction was particularly significant with respect to F2. The size of the vowel space was significantly smaller in the hearing-impaired children. Finally, a smaller acoustic differentiation between the vowels was observed in children with hearing impairment. The results show that even after 5 years of device use, the acoustic characteristics of the vowels in hearing-assisted children remain significantly different as compared to their NH peers.


[Intracardiac hemodynamics after closure of atrial septal defects with the endovascular device, Amplatzer septal occluder].
Changed size and forms of the right and left ventricles of the heart and intraventricular hemodynamics were studied in 5 patients after closing the defects of the interventricular septum with an intravascular device. It was found that after occlusion of the defects by the safe and atraumatic endovascular device the right ventricular myocardium of the heart relaxed, the stroke index became less, the ejection fraction became greater, the diastolic diameter of the right ventricle decreased, and of the left ventricle increased. The indices of the regional contraction function of the tri-cuspid and output sectors of the input part and external and pulmonary output sectors of the right ventricle were found to normalize.


No relevant midbrain atrophy in Parkinson's disease.
To investigate whether significant midbrain atrophy is present in Parkinson's disease (PD), and if so, whether it can be used as a marker of striatal dopaminergic degeneration. In total, 150 PD patients and 155 controls were scanned with both brain dopamine transporter (DAT) [123 I]FP-CIT SPECT and 1.5T MRI. Midbrain atrophy was measured from sagittal MRIs using the midbrain-to-pons ratios. Both striatal region-of-interest-based (Brass) and striatal and extrastriatal voxel-by-voxel-based DAT binding (SPM8) were investigated in relation to midbrain atrophy. The midbrain-to-pons ratios in PD patients were slightly lower than those in the controls (mean 0.59 vs 0.61, P < 0.05). The ratios did not significantly correlate with striatal or extrastriatal [123 I]FP-CIT uptake in controls or patients with PD. Mild midbrain atrophy is present in PD and can be detected with MRI. However, the midbrain atrophy in PD is not associated with the level of striatal dopaminergic dysfunction, and midbrain measurements therefore cannot be used as a clinically useful predictor of dopamine function.


Evaluating the Utility of Toxicologic Analysis in Pediatric Out-of-Hospital Cardiac Arrest.
The cause of a pediatric out-of-hospital cardiac arrest (OHCA) may go unexplained in the emergency department setting but can be secondary to a toxicologic etiology. It is unclear how toxicologic screens are used in the postarrest period after a pediatric OHCA. The primary objectives are to describe 1) when the toxicology screen (urine and serum) is used, 2) patient characteristics, and 3) toxicology screen results. We hypothesized that toxicology screens are frequently used but that positive results are uncommon. This was a retrospective study of pediatric OHCA patients admitted to the Penn State Health Children's Hospital pediatric intensive care unit as transfers from the emergency department between January 1, 2011 and May 31, 2018. We reviewed the electronic health record and evaluated for toxicology screen completion, patient characteristics, and toxicology screen results. One hundred forty-one patients had a pediatric OHCA. Sixty-three (44.7%) patients did not have a toxicology screen completed. A toxicology screen had a higher completion rate for children >11 years of age (n = 26 [78.8%]; p = 0.0024), and in unwitnessed arrests (n = 48 [66.7%]; p = 0.0052). Four cases (5.1%) revealed the presence of substances that were not administered by a medical provider or were illicit. Our study found that in pediatric OHCA, toxicologic screens were completed but were not routinely sent in our institution. There may be factors such as clinician bias or the severity of a patient's illness that impact the approach to toxicologic screening in pediatric OHCA. In addition to the history and physical examination, emergency physician and pediatric intensivists should consider routinely sending toxicologic screens to assist in uncovering any accidental or malicious explanation for the event.


High-Dose Chemotherapy in Children with Newly Diagnosed Medulloblastoma.
High-dose chemotherapy with stem cell rescue has been used as an adjuvant therapy or as salvage therapy to treat pediatric patients with brain tumors, and to avoid deleterious side effects of radiotherapy in infants and very young children. Here, we present the most recent trials using high-dose chemotherapy regimens for medulloblastoma in children, and we discuss their contribution to improved survival and describe their toxicity profile and limitations.


Determination of the prevalence of whole blood taurine in Irish wolfhound dogs with and without echocardiographic evidence of dilated cardiomyopathy.
Taurine plays an important role in maintaining myocardial function. Irish wolfhound dogs (IW) are at risk for dilated cardiomyopathy (DCM), but a relationship between whole blood taurine (WBT) deficiency and DCM has not been established. Our aim was to determine prevalence of WBT deficiency in IW with and without DCM and assess its association with diet. 115 privately owned IW. Whole blood taurine was measured in IW that received cardiovascular examination. Dietary history was recorded; crude protein and energy intake were estimated. Forty-nine (42.6%) had DCM; 66 (57.4%) had no DCM. Dogs with DCM were older ([median; inter-quartile range or IQR] 5.3; 4.3, 6.2 years) than dogs without heart disease (3; 2, 4 years; P < 0.001). There was no significant relationship between WBT concentration and age (P = 0.64). Whole blood taurine was severely reduced (<130 nmol/mL) in 8 dogs (4 with and 4 without DCM) and moderately reduced (130-179.9 nmol/mL) in 32 dogs (12 with DCM and 20 without DCM). Follow up of dogs without DCM revealed that a higher proportion of dogs with any degree of WBT deficiency developed DCM later compared to dogs with normal WBT (P < 0.001). Whole blood taurine deficiency occurred in IW with and without DCM. Based on taurine measurement on a single occasion, there was no clear relationship between low WBT and presence of DCM in this population. Regardless of WBT, DCM affected predominantly older dogs, suggesting a relatively late onset disease in the IW.


Perforator flap terminology: update 2002.
The first terminology consensus on perforator flaps, reached in 2001, was reviewed and further simplified following the Sixth International Course on Perforator Flaps.


Liver Transplantation for Acute Liver Failure During Pregnancy: Case Report and Literature Review.
Liver failure during pregnancy is a rare and highly fatal condition frequently resulting from the patient's pre-existing chronic liver disease. There is no established protocol for managing this condition, and physicians tend to adopt a conservative approach due to concerns about the impact of liver transplantation on fetal development. However, conservative treatment may exacerbate the condition, increasing the risk of fetal and miscarriage deaths. In this study, we present a case of acute-on-chronic liver failure resulting from chronic hepatitis during the 19th week of gestation. Despite undergoing conservative treatment, the fetus tragically died, but the patient achieved an excellent clinical outcome after emergency liver transplantation. We also conducted a systematic review of the literature on liver transplantation during pregnancy from the last decade, examining the causes of liver failure and transplantation during pregnancy. Our research complied with the principles outlined in the Declaration of Helsinki and the Istanbul Declaration.


Expression, purification and characterization of recombinant human beta-amyloid 1-42 in Pichia pastoris.
The human peptide rhA beta(1-42) was effectively produced through a novel expression system and purification procedure. The peptide rhA beta(1-42) was successfully expressed in Pichia pastoris, the methylotrophic yeast that has never been used as host. The cDNA encoding full-length hA beta(1-42) was synthesized with yeast bias codons and cloned into the pPICZ alpha A vector in frame with the yeast alpha-factor secretion signal under the transcriptional control of the AOX1 promoter and integrated into the secreting expression organism P. pastoris strain X33. Production of rhA beta(1-42) through fermentation was further optimized and scaled up in an 80 L fermentor. Secreted rhA beta(1-42) was purified using a two-step purification scheme: SP Sepharose ion exchange chromatography and source 30 RPC. The purification procedure is fast and efficient and reached a recovery of >93% without loss of activity. The purified rhA beta(1-42) was confirmed by Western blotting analysis and N-terminals amino sequencing analysis. This efficient and cost-effective expression system facilitates large-scale production and purification for recombinant rhA beta(1-42).


The relationship between number of interphase nors and nor-bearing chromosomes in non-Hodgkin's lymphoma.
The ribosomal genes (located on the acrocentric chromosomes 13-15, 21-22) may be identified by their silver stained gene products, i.e. NOR related proteins. The NOR bearing chromosome activity can be observed at metaphase with the potential for all ten chromosomes to be positively stained. On the other hand, during interphase they fuse so that eventually only a single silver positive structure is seen in resting normal cells. Investigations of histopathological sections of non-Hodgkin's lymphoma (NHL) have demonstrated a correlation between the numbers of interphase NORs and the grade of tumour. There is generally a higher number of interphase in high-grade, and a lower number in low-grade tumours. This histopathological and cytogenetic study of 13 patients with NHL shows that the higher numbers of interphase NORs in the high-grade tumours is not necessarily a reflection of increased numbers of NOR-bearing chromosomes. Examples were found of high-grade neoplasms, showing the expected high numbers of interphase NORs, but not an increased number of NOR-bearing chromosomes. Conversely, some low-grade tumours, with the expected low number of interphase NORs, had increased numbers of NOR-bearing chromosomes. Our conclusion is that the interphase NOR number is related to factors other than chromosome numbers. We suggest that NOR numbers at interphase may be related to cell turnover. This is supported by previous investigations using DNA flow cytometry and the monoclonal antibody Ki67.


Pre-mRNA with a trimethylguanosine cap structure can be spliced efficiently in vitro.
A chemically synthesized 2,2,7-trimethylguanosine cap (TMG-cap) analogue was added to an in vitro pre-mRNA splicing system. A slight inhibition of the splicing was observed, but it was much less remarkable than that by a 7-methylguanosine cap analogue. This result does not favor a direct role of the TMG-cap structure of small nuclear RNAs in pre-mRNA splicing, and suggests that the TMG-cap does not effectively interact with the 7-methylguanosine cap binding factors. Using the SP6 in vitro transcription system and a TMG-cap analogue, we prepared a beta-globin transcript which has a TMG-cap at the 5' end. This TMG-capped transcript was spliced accurately and efficiently in vitro.


Reconstruction of distal fibula with osteoarticular allograft after tumor resection.
Involvement of distal fibula by benign aggressive and malignant tumors usually necessitates resection of the involved segment of fibula. Numerous techniques have been proposed to reconstruct the ankle joint after this procedure, which can result in complications. We introduce reconstruction of ankle joint by fibular osteoarticular allograft. Reconstruction of the distal fibula after wide resection of tumor was carried out in four patients. There were two cases of Ewing sarcoma, one case of osteosarcoma and one giant cell tumor. After wide resection of tumor, we reconstructed the lateral side of the ankle joint by osteoarticular fibular allograft, which was applied and internally fixed with semitubular plate and screws. In the follow up period, we did assessment of complications, pain and ankle joint instability. The mean age of our patients was 24.2 years (12-31). The mean follow-up was 3.2 years (1.5-6.7). In follow up visits there were no signs of infection or wound healing problems. Union was achieved in all patients. In cases of benign aggressive and malignant tumors involving the distal fibula, we can recommend resection of the distal fibula and reconstruction of the ankle with osteoarticular allograft of the distal fibula. Case series level IV.


[Histologic and biochemical studies of the uteri of ovariectomized gilts during the first two months of pregnancy after different hormone substitutions. 2. Effects of reducing the progesterone dosage on the survival rate of embryos and the structure of the placenta and the influence of oral administration of norgestrel].
Studies were conducted into ovariectomised gilts in early gravidity, with the view to finding out if reduction of daily progesterone doses had an impact upon embryo survival rate and on the structure of the placenta and if experimentally induced progesterone deficit could be offset by oral administration of norgestrel, a synthetic progestagen. Embryo survival were found to drop from 80 to 26.1 percent in response to reduced progesterone dosage from 120 mg to 40 mg, average numbers of living embryos per animal being 9.6 or 3.0. Oral administration of 12 mg of norgestrel, in addition to injection of 40 mg of progesterone, enhanced the survival rate to 64.6 percent, the average number of living embryos coming to 8.9. Reduction of progesterone doses compared to animals with sufficient progesterone supply. Depressed the beginning decrease of the thickness endometrium and surface epithelium oedematisation of the endometrial stroma was mitigated, and subepithelial hyperaemia disappeared altogether. The secretory activity of uterine glands declined, and so did the endometrial activities of acid and alkaline phosphatases. Administration of norgestrel proved helpful in substantive removal of manifestations observed in the progesterone deficit group.


A Successful Second Stem Cell Transplantation in Fanconi Anemia: A Case Report.
The aim was to describe a case of successful second stem cell transplantation in Fanconi anemia (FA). A 7.5-year-old boy with FA was admitted for stem cell transplantation. The first matched sibling allogeneic bone marrow transplantation was done in January 2005. Until August 2008, the patient's blood count had been decreasing and the patient depended on transfusions, and anti-A Ab was detected (donor blood GP is A RH+). Therefore, the second matched sibling allogeneic peripheral blood stem cell transplantation was performed on November 27, 2008 with anti-thymocyte globulin alone as a second transplant conditioning. The child has been disease-free for 5 years. Anti-thymocyte globulin alone as a second transplant conditioning may give first transplant failure FA patients the opportunity for a long-term disease-free survival.


A highly conserved segmental duplication in the subtelomeres of Plasmodium falciparum chromosomes varies in copy number.
Segmental duplications (SD) have been found in genomes of various organisms, often accumulated at the ends of chromosomes. It has been assumed that the sequence homology in-between the SDs allow for ectopic interactions that may contribute to the emergence of new genes or gene variants through recombinatorial events. In silico analysis of the 3D7 Plasmodium falciparum genome, conducted to investigate the subtelomeric compartments, led to the identification of subtelomeric SDs. Sequence variation and copy number polymorphisms of the SDs were studied by DNA sequencing, real-time quantitative PCR (qPCR) and fluorescent in situ hybridization (FISH). The levels of transcription and the developmental expression of copy number variant genes were investigated by qPCR. A block of six genes of >10 kilobases in size, including var, rif, pfmc-2tm and three hypothetical genes (n-, o- and q-gene), was found duplicated in the subtelomeric regions of chromosomes 1, 2, 3, 6, 7, 10 and 11 (SD1). The number of SD1 per genome was found to vary from 4 to 8 copies in between different parasites. The intragenic regions of SD1 were found to be highly conserved across ten distinct fresh and long-term cultivated P. falciparum. Sequence variation was detected in a approximately 23 amino-acid long hypervariable region of a surface-exposed loop of PFMC-2TM. A hypothetical gene within SD1, the n-gene, encoding a PEXEL/VTS-containing two-transmembrane protein was found expressed in ring stage parasites. The n-gene transcription levels were found to correlate to the number of n-gene copies. Fragments of SD1 harbouring two or three of the SD1-genes (o-gene, pfmc-2tm, q-gene) were also found in the 3D7 genome. In addition a related second SD, SD2, of approximately 55% sequence identity to SD1 was found duplicated in a fresh clinical isolate but was only present in a single copy in 3D7 and in other P. falciparum lines or clones. Plasmodium falciparum carries multiple sequence conserved SDs in the otherwise highly variable subtelomeres of its chromosomes. The uniqueness of the SDs amongst plasmodium species, and the conserved nature of the genes within, is intriguing and suggests an important role of the SD to P. falciparum.


A laterally rotating surgical device attached to the operating table.
A laterally rotating surgical device that can be attached to conventional operating tables is described. The position of the patient can be safely and rapidly changed from a supine to a right or left lateral position of 60 degrees. This device is beneficial for performing operations using a thoracoabdominal approach such as is used in cases of resection of gastroesophageal cancer.


Portal gas in a diabetic patient under pharmacotherapy with an alpha-glucosidase inhibitor: report of a case with emphasis on US findings.
Portal gas is relatively rare, and its relation to ischemic bowel diseases has been emphasized. We report the case of a 56-year-old diabetic man under pharmacotherapy with an alpha-glucosidase inhibitor in which portal gas was incidentally detected by ultrasound (US). It showed multiple echo spots moving in the portal vein. Doppler signals confirmed them to be bidirectional and spiky, which immediately led to the diagnosis of portal gas. A change in oral antidiabetic drug caused the portal gas to disappear. We stress the usefulness of US and Doppler US for detecting and diagnosing portal gas. Although detailed analysis of a larger series of diabetic patients under this therapy is needed to draw a definitive conclusion, our observation merits attention. We also briefly review the literature.