A standardized extract of Asparagus officinalis stem prevents reduction in heat shock protein 70 expression in ultraviolet-B-irradiated normal human dermal fibroblasts: an in vitro study.
Heat shock protein 70 (HSP70) exhibits protective effects against ultraviolet (UV)-induced premature skin aging. A standardized extract of Asparagus officinalis stem (EAS) is produced as a novel and unique functional food that induces HSP70 cellular expression. To elucidate the anti-photoaging potencies of EAS, we examined its effects on HSP70 expression levels in UV-B-irradiated normal human dermal fibroblasts (NHDFs). NHDFs were treated with 1 mg/mL of EAS or dextrin (vehicle control) prior to UV-B irradiation (20 mJ/cm2). After culturing NHDFs for different time periods, HSP70 mRNA and protein levels were analyzed using real-time polymerase chain reaction and western blotting, respectively. UV-B-irradiated NHDFs showed reduced HSP70 mRNA levels after 1-6 h of culture, which were recovered after 24 h of culture. Treatment with EAS alone for 24 h increased HSP70 mRNA levels in the NHDFs, but the increase was not reflected in its protein levels. On the other hand, pretreatment with EAS abolished the UV-B irradiation-induced reduction in HSP70 expression at both mRNA and protein levels. These results suggest that EAS is capable to preserve HSP70 quantity in UV-B-irradiated NHDFs. EAS exhibits anti-photoaging potencies by preventing the reduction in HSP70 expression in UV-irradiated dermal fibroblasts.


Genetic features shared by Mycobacterium tuberculosis strains involved in microevolution events.
Microevolved Mycobacterium tuberculosis (MTB) clonal variants from a parental strain can emerge within a single patient infection and during transmission events. Genotypic rearrangements may involve functional changes conferring advantages to favor strain adaptation to the host. In the present study, we analyzed in depth some genotypic characteristics of a strain with a high tendency to microevolve that generated 6 clonal variants during transmission of sequential hosts. In order to identify genetic features potentially associated to microevolution in MTB, we analyzed 56 3R genes and the IS6110 insertion sites from this strain and identified an SNP in alkA and an IS6110 copy located upstream of a transposase (Rv0755A). These markers could be involved in mechanisms leading to genotypic variation. Both features were shared by strains from our collection that were also involved in microevolution, suggesting their putative association with these events.


Cofactor squelching: Artifact or fact?
Cofactor squelching is the term used to describe competition between transcription factors (TFs) for a limited amount of cofactors in a cell with the functional consequence that TFs in a given cell interfere with the activity of each other. Since cofactor squelching was proposed based primarily on reporter assays some 30 years ago, it has remained controversial, and the idea that it could be a physiologically relevant mechanism for transcriptional repression has not received much support. However, recent genome-wide studies have demonstrated that signal-dependent TFs are very often absent from the enhancers that are acutely repressed by those signals, which is consistent with an indirect mechanism of repression such as squelching. Here we review these recent studies in the light of the classical studies of cofactor squelching, and we discuss how TF cooperativity in so-called hotspots and super-enhancers may sensitize these to cofactor squelching.


Defaults, context, and knowledge: alternatives for OWL-indexed knowledge bases.
The new Web Ontology Language (OWL) and its Description Logic compatible sublanguage (OWL-DL) explicitly exclude defaults and exceptions, as do all logic based formalisms for ontologies. However, many biomedical applications appear to require default reasoning, at least if they are to be engineered in a maintainable way. Default reasoning has always been one of the great strengths of Frame systems such as Protégé. Resolving this conflict requires analysis of the different uses for defaults and exceptions. In some cases, alternatives can be provided within the OWL framework; in others, it appears that hybrid reasoning about a knowledge base of contingent facts built around the core ontology is necessary. Trade-offs include both human factors and the scaling of computational performance. The analysis presented here is based on the OpenGALEN experience with large scale ontologies using a formalism, GRAIL, which explicitly incorporates constructs for hybrid reasoning, numerous experiments with OWL, and initial work on combining OWL and Protégé.


Structural and functional characterization of the human decorin gene promoter. A homopurine-homopyrimidine S1 nuclease-sensitive region is involved in transcriptional control.
Decorin is a leucine-rich, chondroitin/dermatan sulfate proteoglycan which binds collagen and growth factors. We have recently completed the genomic organization of human decorin and discovered two alternatively spliced leader exons, designated exon Ia and Ib, in the 5'-untranslated region. Initial analysis of the sequences upstream to these two exons showed that promoter Ia contained only two GC boxes while promoter Ib contained a CAAT and two TATA boxes in close proximity to the transcription start site. To determine if these 5'-flanking sequences exhibited promoter activity, chimeric chloramphenicol acetyltransferase expression plasmids containing the promoter region of either exon Ia or Ib were transfected into HeLa and MG-63 osteosarcoma cells. The results showed that only the region flanking exon Ib was functional. In vitro transcription assay generated two transcripts of 92 and 82 base pairs (bp) indicating that both TATA boxes could be used. Using stepwise 5' deletion analysis we found that the minimum promoter region at -140 bp from the transcription start site, which contained only the CAAT and the two TATA boxes, exhibited strong promoter activity. When a larger construct containing an additional 800 bp of upstream region was tested, a significant increase in transcriptional activity was observed. Interestingly, this promoter region contained several putative binding sites for ubiquitous factors (AP1, AP5, and NF-kappa B) and for transforming growth factor-beta and a 150-bp homopurine/homopyrimidine element with several mirror repeats. When contained in a supercoiled plasmid, this sequence exhibited sensitivity to endonuclease S1, an enzyme that preferentially digests single-stranded DNA. Precise S1 mapping, obtained by direct sequencing of nine distinct S1-generated clones, revealed that in all cases the borders of the sensitive sequence resided within the pur/pyr segment. We propose that this region of the promoter could adopt an intramolecular hairpin triplex structure in vivo and may play a role in the chromatin organization at the decorin gene locus. In addition, this region was able to up-regulate a minimal heterologous promoter in transient transfection assays. The results show that the structure of the decorin gene promoter is different from that of any other proteoglycan promoter characterized so far and indicate that the pur/pyr segment plays a role in the regulation of gene transcription.


Assessment of the relationships between genetic determinants of thyroid functions and bipolar disorder: A mendelian randomization study.
Thyroid functions (TFs) have been implicated in the initiation and propagation of psychiatric disorders. Observational studies have shown associations of TFs with psychiatric disorders. However, the relationship between TFs and psychiatric diseases were still unclear. Genetic instruments for 6 TF-realted indexes, including free thyroxine (FT4), triiodothyronine (FT3):FT4 ratio, thyrotropin (TSH), thyroid peroxidase antibodies (TPOAb) concentration, hypothyroidism, and hyperthyroidism, were obtained from several genome-wide association studies (GWASs). Their associations with BD were evaluated using Psychiatric Genomics Consortium (PGC) datasets (41,917 cases and 371,549 controls). All GWAS summary statitics were from European ancestry. Mendelian randomization (MR) estimates from each genetic instrument were combined using inverse variance weighted (IVW) meta-analysis, with complementary methods (eg, weighted median and MR Egger). We also multiple sensitivity analyses to examine horizontal pleiotropy and heterogeneity. Genetically predicted level of FT4 was significantly associated with BD (odds ratio (OR)=0.89, 95% confidence interval (CI): 0.83-0.95; P=4.65 × 10-3), survived after the Bonferroni correction (P<0.05/6=0.008). Consistent directional effects for all sensitivity analyses were observed in the weighted median and MR Egger methods. Furthermore, our sensitive test suggested no significant horizontal pleiotropy (intercept=-0.01, P=0.12) and no notable heterogeneity (Q = 29.9; P=0.09). However, other TF indexes (FT3:FT4 ratio [OR=1.24, P=0.10], TSH [OR=1.01, P=0.61], TPOAb concentration [OR=1.20, P=0.54], hypothyroidism [OR=1.00, P=0.91], and hyperthyroidism [OR=0.99, P=0.57]) were not associated with BD. Our results provide further evidence that higher FT4 level is associated with a reduced risk of BD, and suggest the importance of FT4 level in BD risk assessment and potential therapeutic targets development.


Human fetal neural stem cells in rat brain: effects of preculturing and transplantation.
The fate of human fetal stem/progenitor cells transplanted into rat brain depends on conditions of preculturing (long or short) and state and site of transplantation. Human nestin-positive stem cells cultured according to the short protocol did not migrate into hypoxic and normal brain after transplantation, but actively migrated in damaged spinal cord. After transplantation of long-cultured cells into the brain mainly committed neuroblasts and solitary nestin-positive cells migrated from the site of transplantation into the brain.


Comparison of trace element concentrations in livers of diseased, emaciated and non-diseased southern sea otters from the California coast.
Infectious diseases have been implicated as a cause of high rates of adult mortality in southern sea otters. Exposure to environmental contaminants can compromise the immuno-competence of animals, predisposing them to infectious diseases. In addition to organic pollutants, certain trace elements can modulate the immune system in marine mammals. Nevertheless, reports of occurrence of trace elements, including toxic heavy metals, in sea otters are not available. In this study, concentrations of 20 trace elements (V, Cr, Mn, Co, Cu, Zn, Rb, Sr, Mo, Ag, Cd, In, Sn, Sb, Cs, Ba, Hg, Tl, Pb, and Bi) were measured in livers of southern sea otters found dead along the central California coast (n=80) from 1992 to 2002. Hepatic concentrations of trace elements were compared among sea otters that died from infectious diseases (n=27), those that died from non-infectious causes (n=26), and otters that died in emaciated condition with no evidence of another cause of death (n=27). Concentrations of essential elements in sea otters varied within an order of magnitude, whereas concentrations of non-essential elements varied by two to five orders of magnitude. Hepatic concentrations of Cu and Cd were 10- to 100-fold higher in the sea otters in this study than concentrations reported for any other marine mammal species. Concentrations of Mn, Co, Zn, and Cd were elevated in the diseased and emaciated sea otters relative to the non-diseased sea otters. Elevated concentrations of essential elements such as Mn, Zn, and Co in the diseased/emaciated sea otters suggest that induction of synthesis of metallothionein and superoxide dismutase (SOD) enzyme is occurring in these animals, as a means of protecting the cells from oxidative stress-related injuries. Trace element profiles in diseased and emaciated sea otters suggest that oxidative stress mediates the perturbation of essential-element concentrations. Elevated concentrations of toxic metals such as Cd, in addition to several other organic pollutants, may contribute to oxidative stress-meditated effects in sea otters.


Separating the wheat from the chaff-COVID-19 in a German emergency department: a case-control study.
COVID-19 pandemia is a major challenge to worldwide health care systems. Whereas the majority of disease presents with mild symptoms that can be treated as outpatients, severely ill COVID-19 patients and patients presenting with similar symptoms cross their ways in the emergency department. Especially, the variety of symptoms is challenging with primary triage. Are there parameters to distinguish between proven COVID-19 and without before? How can a safe and efficient management of these inpatients be achieved? We conducted a retrospective analysis of 314 consecutive inpatient patients who presented with possible symptoms of COVID-19 in a German emergency department between March and April 2020 and were tested with a SARS-Cov-2 nasopharyngeal swab. Clinical parameters, Manchester Triage System categories, and lab results were compared between patients with positive and negative test results for SARS-Cov-2. Furthermore, we present the existing COVID-19 workflow model of the university hospital in Essen which proved to be efficient during pandemia. Forty-three of the 314 patients (13.7%) were tested positive for COVID-19 by SARS-Cov-2 nasopharyngeal swab. We did not find any laboratory parameter to distinguish safely between patients with COVID-19 and those with similar symptoms. Dysgeusia was the only clinical symptom that was significantly more frequent among COVID-19 patients. Dysgeusia seems to be a typical symptom for COVID-19, which occurred in 14% of our COVID-19 patients. However, no valid parameters could be found to distinguish clinically between COVID-19 and other diseases with similar symptoms. Therefore, early testing, a strict isolation policy, and proper personal protection are crucial to maintain workflow and safety of patients and ED staff for the months to come. German Clinical Trials registry, DRKS00021675.


[Peritoneovenous shunt complicating a glomerulonephritis associated with bacteremia. Description of a clinical case].
We report the case of a patient who exhibited a complication following implantation of the LeVeen valve. This is the first example of a delayed complication in our experience, occurring approximately 3 years after the implantation. Biochemical and clinical data showed bacteremia due to meningococci resulting from the valve (and this was subsequently confirmed by microbiological data).


The time interval between laparoscopic tubal ligation and frozen-thawed embryo transfer does not affect the reproductive outcomes.
Hydrosalpinx may decrease implantation and pregnancy rates after embryo transfer. Laparoscopic tubal ligation after embryo freeze and before frozen-thawed embryo transfer (FET) is effective at improving reproductive outcomes for hydrosalpinx patients. This study is to find out the optimal interval between laparoscopic tubal ligation and FET. We retrospectively analyzed 259 infertile women who performed laparoscopic tubal ligation for embryo freeze and FET. Participants were divided into three groups, based on the interval between laparoscopic tubal ligation and FET. Group I: <30 days; Group II: 31- 60 days; Group III: >60 days. Outcomes of cleavage-stage and blastocyst-stage embryo FET were analyzed respectively. There was no significant difference in clinical pregnancy rate, live birth rate, implantation rate, biochemical pregnancy rate, ectopic pregnancy rate, miscarriage rate and preterm birth rate among the three groups, in both cleavage-stage and blastocyst-stage embryo FET cycles. In cleavage-stage embryo FET cycle, singleton gestational age was significantly younger in group III (38.11 ± 2.28 weeks) compared with group I (39.29 ± 1.06 weeks, P = 0.001) and group II (38.96 ± 1.05, P = 0.026). Singleton birth weight was significantly heavier in group II (3.65 ± 0.32 Kg) compared with group I (3.38 ± 0.29 Kg, P = 0.001) and group III (3.35 ± 0.60 Kg, P = 0.004). Twin birth weight was significantly heavier in group III (2.72 ± 0.43 Kg) compared to group I (2.23 ± 0.67 Kg, P = 0.002). In blastocyst-stage embryo FET cycles, twin gestational age was significantly younger in group II (34.07 ± 3.18 weeks) compared with group I (35.56 ± 2.27 weeks, P = 0.049) and group III (36.50 ± 1.47 weeks, P = 0.005). Twin birth weight was significantly heavier in group III (2.71 ± 0.39 Kg) compared to group II (2.39 ± 0.67 Kg, P = 0.009). The duration of the interval between laparoscopic tubal ligation and FET does not affect the reproductive outcomes; however, it may affect the neonate outcomes to some extent.


Eosinophilic peritonitis in children on chronic peritoneal dialysis.
Eosinophilic peritonitis (EP) is sometimes difficult to distinguish from bacterial peritonitis (BP) at onset, as they are often overlapping. Previous reports show EP occurs more frequently in infants, although the reason is unknown. The study population was 77 pediatric patients receiving chronic peritoneal dialysis (PD) in our center. We compared clinical and laboratory data at onset of EP with those of BP. We also investigated age distribution at onset of EP and PD-related surgery. Eleven patients developed EP (18 episodes) and 19 patients developed BP (38 episodes). EP patients showed lower rate of cloudy dialysate (44.4% vs. 74.4%; p = 0.04), lower rate of fever (38.9% vs. 56.4%), lower frequency of abdominal pain (16.7% vs. 38.5%), higher peripheral blood eosinophil counts (/μL) (514 vs. 160; p < 0.001), and lower serum C-reactive protein level (mg/dL) (0.4 vs. 4.7; p < 0.001) than BP patients. Thirteen EP events were observed after 169 surgical interventions. Age at surgery-related EP was similar to age at surgery without EP (2.6 vs. 2.1; p = 0.65). There was no significant difference in postoperative EP occurrence between groups <2 years and ≥ 2 years (6.2% vs. 9.1%; p = 0.48). However, infants received more operations than older children. Clinical symptoms in children and laboratory data of EP in children were less severe than those of BP. As incidence of postoperative EP did not differ by age, we speculate that higher incidence of EP in infants might be associated with higher incidence of surgery, although further validation is necessary.


Penile plication without degloving enables effective correction of complex Peyronie's deformities.
To present our initial experience with extended plication repair for men with severe and/or biplanar penile curvature. A review of men who underwent plication repair for complex penile deformity (biplanar curvature or curvature ≥ 60°) was performed. All patients underwent tunical plication via a 2-cm penoscrotal incision mobilized distally along the penile shaft without degloving. Angle of curvature, direction(s), stretched penile length (SPL), and number of sutures were recorded. Among 102 patients treated with plication surgery, 43 (44%) had complex penile deformity. Among 11 men with biplanar curvature, median angle in the primary plane of curvature was corrected from 45° to 10° and secondary plane was corrected from 35° to 5° using an average of 7 sutures (5° correction per suture). Among 32 men with severe curvature, median angle of curvature was corrected from 70° to 15° using an average of 11 sutures (6° correction per suture). SPL was unchanged in 29 (69%), increased an average of 0.65 cm in 7 (16%), and decreased 0.5 cm in only 6 (14%) patients. At a mean follow-up of 15.3 months, repeat plication was required in 2 patients and 2 required penile prosthesis. Penile plication without degloving appears to be safe and effective for correction of complex penile curvature without significant impact on penile length.


[Histologic detection of cytomegalovirus and its application in gastrointestinal cytomegalovirus infection].
To investigate the clinical and pathologic features of gastrointestinal cytomegalovirus (CMV) disease, and the histological detection of CMV. Forty-one gastrointestinal tissues were obtained from 35 patients suspected for gastrointestinal CMV disease. The pathologic changes of these specimens were evaluated by immunohistochemistry and in situ hybridization for CMV, and these were compared to serologic detection methods. CMV was detected in 23/41 tissues. There were 32 serologic CMV DNA tests at the time of biopsies or operations. Compared with histological detection, the sensitivity and specificity of serologic CMV DNA tests were 73.7% and 69.2% respectively. Gastrointestinal histological detection of CMV has important clinical application. HE staining combined with immunohistochemistry and/or in situ hybridization detection is a reliable method to detect CMV.


Cleavage of the selective autophagy receptor SQSTM1/p62 by the SARS-CoV-2 main protease NSP5 prevents the autophagic degradation of viral membrane proteins.
Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has caused the coronavirus disease 2019 (COVID-19) global pandemic. Omicron, a new variant of SARS-CoV-2, has the characteristics of strong transmission and pathogenicity, short incubation period, and rapid onset progression, and has spread rapidly around the world. The high replication rate and intracellular accumulation of SARS-CoV-2 are remarkable, but the underlying molecular mechanisms remain unclear. Autophagy acts as a conservative cellular defence mechanism against invading pathogens. Here, we provide evidence that the main protease of SARS-CoV-2, NSP5, effectively cleaves the selective autophagy receptor p62. NSP5 targets p62 for cleavage at glutamic acid 354 and thus abolishes the capacity of p62 to mediate selective autophagy. It was further shown that p62 specifically interacted with ubiquitinated SARS-CoV-2 M, the viral membrane protein, to promote its autophagic degradation. In the presence of NSP5, p62-mediated autophagic degradation of the M protein was inhibited. The cleaved products of p62 also cannot facilitate the degradation of the M protein. Collectively, our findings reveal that p62 is a novel host target of SARS-CoV-2 NSP5 and suggest that selective autophagy targets viruses and potential strategies by which the virus evades autophagic clearance. Our results may provide new ideas for the development of anti-COVID-19 drugs based on autophagy and NSP5.


Quantitative saccharide release of hydrothermally treated flours by validated salivary/pancreatic on-surface amylolysis (nanoGIT) and high-performance thin-layer chromatography.
The susceptibility of hydrothermally treated flour products to amylolysis was studied. The human salivary α-amylase and porcine pancreatin enzyme mixture containing α-amylase were used on-surface to investigate the release of glucose, maltose, and maltotriose. On the same adsorbent surface (all-in-one), their high-performance thin-layer chromatography separation and detection via selective chemical derivatization was performed. For the first time, the all-in-one nanoGIT system was studied quantitatively and validated for the simulated static oral and intestinal on-surface amylolysis of ten different hydrothermally treated flours and soluble starch. Differences were detected in the digestibility of refined and whole flours from wheat, spelt, and rye as well as from einkorn, amaranth, emmer, and oat. Amaranth released the lowest amount of saccharides and spelt the highest in both oral and intestinal digestion systems. The results suggest that consumption of whole grain products may be beneficial because of their lower saccharide release, with particular attention to rye.


Neural surveillance of skeletal homeostasis.
Endowed with sympathetic and peptidergic nerves, the vertebrate skeleton is under constant surveillance by the nervous system. In addition to pituitary hormone secretion, centrally regulated sympathetic release, as elegantly demonstrated by Karsenty and colleagues, integrate to control both components of skeletal remodeling, osteoblastic bone formation, and osteoclastic bone resorption (Elefteriou et al., 2005).


Are high cumulative doses of erythropoietin neuroprotective in preterm infants? A two year follow-up report.
Preterm infants are at risk for neurodevelopmental sequelae even in absence of major cerebral lesions. The hypothesis that Human Recombinant Erythropoietin (rEpo) could improve the neurodevelopmental outcome in risk neonates has raised the highest interest in recent years. A group of preterm neonates born at a gestational age ≤ 30 weeks and free from major cerebral lesions or major visual impairment, were included in the study if they had a complete neurologic evaluation for at least 24 months of postmenstrual age. They were assigned to group I in the case they had been treated with rEpo or group II if untreated. The aim was to evaluate whether rEpo, given at the high cumulative doses utilized for hematologic purposes, is able to improve the neurodevelopmental outcome in preterm infants born at a gestational age ≤ 30 weeks. A group of 104 preterm neonates were studied: 59 neonates who received rEpo for 6.9 ± 2.4 weeks at a median cumulative dose of 6300 UI/Kg (6337 ± 2434 UI/Kg), starting at a median age of 4 days and 45 neonates who were born in the period preceding the routine use of rEpo. The neurodevelopmental quotient at 24 month postmenstrual age was assessed utilizing the Griffiths' Mental Developmental Scales. Our results failed to show any difference in the Developmental Quotient at 24 month. Bronchopulmonary dysplasia, minor intraventricular hemorrhages and blood transfusions were the clinical features significantly related to the Developmental Quotient. Our results do not support the hypothesis that rEpo, administered with the schedule utilized for hematologic purposes, improve the neurodevelopmental outcome of preterm neonates, at least those preterm infants free from major impairments.


Extreme plastomes in holoparasitic Balanophoraceae are not the norm.
Balanophoraceae plastomes are known for their highly condensed and re-arranged nature alongside the most extreme nucleotide compositional bias known to date, culminating in two independent reconfigurations of their genetic code. Currently, a large portion of the Balanophoraceae diversity remains unexplored, hindering, among others, evolutionary pattern recognition. Here, we explored newly sequenced plastomes of Sarcophyte sanguinea and Thonningia sanguinea. The reconstructed plastomes were analyzed using various methods of comparative genomics based on a representative taxon sampling. Sarcophyte, recovered sister to the other sampled Balanophoraceae s. str., has plastomes up to 50% larger than those currently published. Its gene set contains five genes lost in any other species, including matK. Five cis-spliced introns are maintained. In contrast, the Thonningia plastome is similarly reduced to published Balanophoraceae and retains only a single cis-spliced intron. Its protein-coding genes show a more biased codon usage compared to Sarcophyte, with an accumulation of in-frame TAG stop codons. Structural plastome comparison revealed multiple, previously unknown, structural rearrangements within Balanophoraceae. For the "minimal plastomes" of Thonningia, we propose a genetic code change identical to sister genus Balanophora. Sarcophyte however differs drastically from our current understanding on Balanophoraceae plastomes. With a less-extreme nucleotide composition, there is no evidence for an altered genetic code. Using comparative genomics, we identified a hotspot for plastome reconfiguration in Balanophoraceae. Based on previously published and newly identified structural reconfigurations, we propose an updated model of evolutionary plastome trajectories for Balanophoraceae, illustrating a much greater plastome diversity than previously known.


Solid phase synthesis of oligomannopeptoids that mimic the concanavalin A-binding trimannoside.
Oligomannopeptoids from the dimer to the hexamer were produced by solid phase synthesis and their abilities to bind to concanavalin A (ConA) were assessed. The assessment indicated similarity between the oligomannopeptoids and the naturally occurring oligomannosides in the enthalpy of the binding and the valence number vs binding strength relationship, encouraging the use of the oligomannopeptoids as oligomannoside mimics.


trans-activation of viral enhancers including long terminal repeat of the human immunodeficiency virus by the hepatitis B virus X protein.
Human hepatitis B virus contains an open reading frame designated X. We have investigated the trans-activating function of the hepatitis B virus X gene in regulating transcriptional control elements. In the HBV genome the major target for X trans-activation is the enhancer element. Further, the X protein stimulates several other viral promoters/enhancers including the long terminal repeats (LTR) of human retroviruses. One of the viral sequences studied in detail is the human immunodeficiency viral (HIV) LTR which is trans-activated by the X protein. Using mutational analysis of the HIV LTR, we show that the NF-kappa B sequences contained within the U3 region are involved in this stimulatory activity. Nuclear run-on analyses support the notion that X-mediated trans-activation occurs at the level of transcription.


Management of pulmonary insufficiency in diaphragmatic hernia using extracorporeal circulation with a membrane oxygenator (ECMO).
Persistent fetal circulation (PFC) causes severe pulmonary insufficiency in patients who have demonstrated adequate lung function following diaphragmatic hernia repair. Patent ductus arteriosus (PDA) ligation corrects this condition, but carries the risk of sudden right ventricular failure. Pharmacologic reversal of PFC may be attempted, and if unsuccessful, prolonged venoarterial bypass becomes necessary to provide effective pulmonary support. PDA ligation can then be performed safely and maturation of the pulmonary vasculature allowed to occur. Pulmonary artery pressure monitoring is essential.


Puerto Rican athletes with special health care needs: an evaluation of oral health status.
Limited data are available regarding the oral health of individuals with disabilities who reside in Puerto Rico. The Special Olympics Special Smiles oral health screening, education, and referral program provides an opportunity to explore the dental status and needs of Special Olympics athletes with intellectual and developmental disabilities. The purpose of this paper was to evaluate the oral health status of Special Olympics athletes participating in an event in Villalba, Puerto Rico in 2004. A total of 161 self-selected athletes with intellectual disabilities were screened by 4 trained and calibrated dentists utilizing the criteria standards of the National Institute of Dental and Craniofacial Research, Special Olympics Special Smiles, and the Centers for Disease Control and Prevention developed for decayed, filled, and missing teeth. Oral hygiene, reports of oral pain, and sociodemographic information were recorded. The statistical test used was the chi-square test to establish associations and logistic regressions. Data were analyzed using EPI-INFO and SPSS programs. Almost 45% of examined athletes (mean age=19 years) had untreated dental decay; 33% had missing teeth, 60% had gingival signs of periodontal disease, and 16% reported dental pain. Results of clinical examinations indicate a population with intellectual disabilities that is in significant need of preventive and restorative oral health needs.


The global prevalence of screen-based disordered eating and associated risk factors among high school students: systematic review, meta-analysis, and meta-regression.
Estimate the prevalence, and associated risk factors, of high school students who are considered at risk for an eating disorder based on screening measures. An electronic search of nine databases was completed from their inception until 1st September 2022. A random-effects meta-analysis was conducted, and confounder (moderator) analyses and meta-regressions examined whether the overall prevalence estimate for of screen-based disordered eating (SBDE) was moderated by student age, BMI, or gender, as well as culture and type of SBDE assessment. The mean estimate of the prevalence of SBDE among high school students (K = 42 (66 datapoints), N = 56282] in the sample of 25 countries was 13% ([95% CI] = 10.0-16.8%, I2 = 99.0%, Cochran's Q p = 0.001). This effect was not moderated by features of the samples such as gender, BMI, or age. Among cultures, non-Western countries had a higher prevalence of SBDE prevalence than Western countries, but the difference was not significant. There was considerable variability in the prevalence estimates as a function of the assessment measure, but no meaningful pattern emerged. The estimated figure of 1 in 8 high school students with SBDE-unmoderated by gender and BMI-stands out as a problem in need of attention from public health officials, psychologists, psychiatrists, pediatricians, parents, and educators. There is a great need for innovative, integrated policy and program development all along the spectrum of health promotion and universal, selective, and indicated prevention. Further research is also needed to validate and refine this estimate by (a) conducting basic research on the accuracy of eating disorder screening measurements in samples ages 14 through 17; (b) examining representative samples in more countries in general and Latin American countries in particular; (c) clarifying the relationships between SBDE and age throughout the different phases of late childhood, adolescence, and emerging adulthood; and (d) investigating whether there are meaningful forms of disordered eating and whether these are associated with variables such as gender, ethnicity, and BMI.


Comparison of three cost effective staining methods for detection of Pneumocystis jirovecii.
Pneumocystis pneumonia due to Pneumocystis jirovecii infection is an emerging health problem not only for HIV-infected patients but also for other immunocompromised patients in many countries. We compared Gomori methenamine silver (GMS), Toluidine Blue O (TBO) and Giemsa staining methods using standard procedures. The sensitivity and specificity of GMS were 100 %. The sensitivity and specificity of TBO were 96 and 100 %, respectively. The sensitivity and specificity of Giemsa stain were 84 and 90 %, respectively. Only GMS had positive and negative predictive values of 100 % while PPV and NPV for TBO were 100 and 90.9 %, and for Giemsa stain were 95.4 and 69.2 %, respectively. Therefore, our results suggest that if TBO or Geimsa stains are used as the primary staining methods in a clinical laboratory, then confirmation with a GMS staining method should be performed to increase the sensitivity and specificity of the final test result.


The THCRE2 site in the rat tyrosine hydroxylase gene promoter is responsive to phorbol ester.
Recently, our laboratory has identified a tyrosine hydroxylase (TH) gene promoter element, THCRE2 (located at -97 to -90), that is required for maximal response to cyclic AMP. In this study we test whether the THCRE2 is responsive to phorbol ester. Rat PC12 cells were transfected with a reporter gene construct, TH(-272/+27)-CAT, which is driven by the first 272 bp of the rat TH gene 5' flanking region. Treatment of transfected cells with 0.1 microM 12-O-tetradecanoylphorbol 13-acetate (TPA) elicited a 5-6-fold increase in TH gene proximal promoter activity. Mutagenesis of the THCRE2 sequence diminished TPA-responsiveness of the TH gene promoter by approximately 50%. Minimal promoter constructs driven by a single copy of the region of the TH gene that encodes the THCRE2 (from -117 to -59) were also responsive to TPA. Our results suggest that the THCRE2 is a phorbol ester-responsive element, as well as a cyclic AMP-responsive element.


DNA-damage processing in a radiation-sensitive mouse cell line.
The induction and repair of radiation-induced DNA damage was assessed in 3 mouse cell lines, including the parental L cell line, a radiation-sensitive, SL3-147 mutant cell line and the H5 revertant to radiation resistance. The yield of neither radiation-induced DNA single- nor double-strand breaks could explain the variable sensitivity of the 3 cell lines. Closure of DNA single-strand breaks proceeded at a similar rate in both the L and SL3-147 cell lines. Closure of DNA double-strand breaks however was significantly slower and less complete in the SL3-147 cell line than in either of the radiation-resistant cell lines. The results are consistent with the increased radiation sensitivity of the SL3-147 cell line resulting from a defect in their ability to repair radiation-induced DNA double-strand breaks.


Suprapubic Lipectomy and Placement of a Three-piece Inflatable Penile Prosthesis.
Penile length loss is often observed in patients with erectile dysfunction and obesity. In patients with a prominent infrapubic fat pad, the excess fat can further reduce perceived penile length.1 As others have reported, concurrent suprapubic lipectomy and placement of a penile implant has been shown to be an effective way of exposing phallic length in this challenging patient population.2,3 OBJECTIVE: In this video, we present surgical technique and initial outcomes of concurrent suprapubic lipectomy and 3-piece inflatable penile prosthesis (IPP) placement in patients that have erectile dysfunction and lipodystrophy of the mons pubis. We retrospectively reviewed an institutional review board-approved prosthetic database of male patients that underwent placement of a 3-piece IPP and concurrent suprapubic lipectomy from 2020 to 2023. Intraoperative complications and postoperative outcomes were collated and analyzed. To perform this operation, an elliptical incision is made around the suprapubic fat pad that extends to the base of the penis inferiorly. Laterally the incision is carried just medial to the inferior margin of the iliac crest. Sharp dissection and electrocautery are used to dissect the fat pad free. A layer of areolar tissue is preserved above the layer of the fascia to prevent seroma formation. For larger blood vessels, vascular clips are used to ensure adequate hemostasis. After removal of the fat pad, placement of the penile prosthesis can proceed as one would through an infrapubic approach. After the device is placed, the tissue flaps based on the external pudendal vessels are inspected for adequate vascularity. Penetrating towel clamps are used to approximate the skin edges from lateral to medial so that the midline closure can be approximated accurately. 0-PDS is used to anchor the flap superiorly into the lower edge of the abdominal fold. The incision is then closed in multiple layers of absorbable suture from lateral to medial. A skin stapler or suture is then used to approximate the skin and the penis is wrapped with a compressive dressing. Nine patients underwent the combined surgery. There were no intraoperative complications. In follow-up, 1 patient developed an infection-related complication that required device removal and immediate replacement. The remaining 8 did well postoperatively with improved dorsal phallic length with no infection or device-related complications. Concurrent suprapubic lipectomy and placement of a 3-piece IPP is a safe and reproducible technique for the treatment of suprapubic lipodystrophy, buried penis, and refractory ED.


Antibacterial, antibiofilm and anti-quorum sensing activities of Muntingia calabura L. leaf extract against Pseudomonas aeruginosa.
The aim of this study was to evaluate the phytochemicals, antioxidant, antibacterial and anti-quorum sensing (QS) properties of methanol extract of Muntingia calabura L. leaves against biofilm-forming strain Pseudomonas aeruginosa. The antioxidant potential of M. calabura methanol leaf extract was evaluated using a radical scavenging assay. Since the findings were so promising, the study aims to screen the phytochemical profiles using qualitative and quantitative approaches and to expand the investigation to anti-QS activity. The antibacterial activity was assessed by determining the minimum inhibitory concentration against pathogenic Gram-negative and Gram-positive biofilm-forming bacterial strains using an agar well diffusion plate method. Muntingia calabura methanol leaf extract was most potent against P. aeruginosa. The QS controlled virulence factors in P. aeruginosa is significantly inhibited by M. calabura leaf extract. The microscopy images revealed a major reduction in P. aeruginosa biofilm formation. Interestingly, M. calabura leaf extract was not toxic to Vero cell line. These findings make M. calabura a strong candidate for new antimicrobial and anti-QS herbal agents.


Regional activation in the rat brain during visceral stimulation detected by c-fos expression and fMRI.
The aim of the study was to determine and compare the areas of brain activated in response to colorectal distention (CRD) using functional magnetic resonance imaging (fMRI) and c-fos protein expression. For fMRI study (3.0 T magnet), anaesthetized rats underwent phasic CRD, synchronized with fMRI acquisition. Stimulation consisted of eight cycles of balloon deflation (90 s) and inflation (30 s), at 40, 60 or 80 mmHg of pressure. For c-fos study two sets of experiments were performed on anaesthetized rats: comparing (A) brain activation in rats with the inserted colorectal balloon (n = 5), to the rats without the balloon (n = 5); and (B) rats with inserted balloon (n = 10), to the rats with inserted and distended balloon (n = 10). The pressure of 80 mmHg was applied for 2 h of 30 s inflation and 90 s deflation, alternating cycles. Functional MRI revealed significant activation in the amygdala, hypothalamus, thalamus, cerebellum and hippocampus. Significant increase in c-fos expression was observed in amygdala and thalamus in the first set of experiments, and hypothalamus and parabrachial nuclei in the second. The two methods are not interchangeable but appeared to be complementary: fMRI was more sensitive, whereas c-fos had much greater resolution.


2,4-Bis(furan-2-yl)-1,5-dimethyl-3-aza-bicyclo-[3.3.1]nonan-9-one.
In the title compound, C18H21NO3, the bicyclic ring system adopts a twin-chair conformation. The two methyl groups attached to the bicycle are in an equatorial orientation for both rings. One of the furan rings is disordered over two orientations with an occupancy ratio of 0.686 (6):0.314 (6). In the crystal, very long N-H⋯O hydrogen bonds connect the mol-ecules into a chain perpendicular to the ac plane.


Utility of expanded anterior column resection versus decompression-alone for local control in the management of carcinomatous vertebral column metastases undergoing adjuvant stereotactic radiotherapy.
With improvements in adjuvant radiotherapy and minimally invasive surgical techniques, separation surgery has become the default surgical intervention for spine metastases at many centers. However, it is unclear if there is clinical benefit from anterior column resection in addition to simple epidural debulking prior to stereotactic body radiotherapy (SBRT). To examine the effect of anterior column debulking versus epidural disease resection alone in the local control of metastases to the bony spine. Retrospective cohort study. Ninety-seven patients who underwent open surgery followed by SBRT for spinal metastases at a single comprehensive cancer center. Local tumor recurrence following surgery and SBRT. Data were collected regarding radiation dose, cancer histology, extent of anterior column resection, and recurrence. Tumor involvement was categorized using the International Spine Radiosurgery Consortium guidelines. Univariable analyses were conducted to determine predictors of local recurrence and time to local recurrence. Among the 97 included patients, mean age was 60.5±11.4 years and 51% of patients were male. The most common primary tumor types were lung (20.6%), breast (17.5%), kidney (13.4%) and prostate (12.4%). Recurrence was seen in 17 patients (17.5%) and local control rates were: 85.5% (1-year), 81.1% (2-year), and 54.9% (5-year). Overall predictors of local recurrence were tumor pathology (p<.01; renal cell carcinoma and colorectal adenocarcinoma associated with poorest PFS) and undergoing anterior column debulking versus epidural decompression-alone (p=.03). Only tumor pathology predicted time to local recurrence (p<.01), though inspection of Kaplan-Meier functions showed superior long-term local control in patients with radiosensitive tumor pathologies, no previous irradiation of the metastasis, and who underwent anterior column resection versus epidural removal alone. Median time to recurrence was 288 days with 100% of lesions showing anterior column recurrence and recurrence in the epidural space. With the increasing shift towards surgery as a neoadjuvant to radiotherapy for patients with spinal column metastases, the role for surgical debulking has become less clear. In the present study, we find that anterior column debulking as opposed to epidural debulking-alone decreases the odds of local recurrence and improves long-term local control.


Extramural vascular invasion is an adverse prognostic indicator of survival in patients with colorectal cancer.
Extramural vascular invasion (EVI) in colorectal cancer is reported to be a stage-independent adverse prognostic factor, and is a core item in the Royal College of Pathologists minimum data set for colorectal cancer histopathology reporting. The detection of EVI is also highly variable amongst pathologists. Our aims were to analyse both the frequency of EVI in colorectal cancer resections, and the effect of EVI on survival, in patients operated on over a 5-year period. A retrospective analysis of patients having potentially curative surgery for colorectal cancer between January 1999 and December 2004. Over 5 years, 378 patients underwent a potentially curative resection. One-hundred seven (28.3%) cancers exhibited EVI, of which 104 (97%) were T3 and T4 tumours. Survival curves with and without EVI, unadjusted for nodal status and T stage, were significantly different (P = 0.0001) with 5-year survivals of 52% and 73% respectively. Survival curves for T3 and T4 tumours stratified with and without EVI also showed significantly different survival distributions (P = 0.007). A significant difference in frequency of EVI year on year was seen (P < 0.001), ranging from 8.5% to 46.7%, whereas the number of T3 and T4 tumours in each year was not significantly different (P = 0.677). EVI is an adverse prognostic indicator for survival in patients undergoing potentially curative resection of colorectal cancer, and the routine requirement of EVI in colorectal cancer histopathology reporting is justified. Optimal specimen preparation, meticulous histopathological analysis and regular auditing of EVI detection rates are essential for the accurate staging of colorectal cancer.


The methylome: approaches for global DNA methylation profiling.
DNA methylation plays a critical role in genome function both in health and disease. Almost 60 years after the discovery of 5-methyl cytosine and approximately 25 years since the discovery that altered DNA methylation plays a role in disease, the first high-resolution DNA methylation profile (or methylome) of any genome--Arabidopsis thaliana--was determined. Although only approximately 20% of the typical size of mammalian genomes, this milestone demonstrated that the methylomes of the human and similarly large genomes are now within reach. Here, we review current and emerging technologies that hold promise to deliver the first mammalian methylome and to facilitate comprehensive profiling of essentially any cell type in the context of development, disease and the environment.


Environments Tune and Select Cellular Diversity.
Technical advances in single-cell sequencing data and their application to greater samples is revealing substantial cell-to-cell variation in expression levels and propagation of this variation between molecules across cells. New quantitative approaches that apply mechanistic and statistical models in a systems-wide approach are illuminating the drivers of phenotypic diversity.


Proposals of Sphingomonas paucimobilis gen. nov. and comb. nov., Sphingomonas parapaucimobilis sp. nov., Sphingomonas yanoikuyae sp. nov., Sphingomonas adhaesiva sp. nov., Sphingomonas capsulata comb. nov., and two genospecies of the genus Sphingomonas.
Based on the partial nucleotide sequence analysis of 16S ribosomal ribonucleic acid (rRNA), presence of unique sphingoglycolipids in cellular lipid, and the major type of ubiquinone (Q10), we propose Sphingomonas gen. nov. with the type species Sphingomonas paucimobilis (Holmes et al, 1977) comb. nov. From the homology values of deoxyribonucleic acid-deoxyribonucleic acid hybridization and the phenotypic characteristics, three new species, Sphingomonas parapaucimobilis, Sphingomonas yanoikuyae, Sphingomonas adhaesiva, and one new combination, Sphingomonas capsulata, are described. S. parapaucimobilis JCM 7510 (= GIFU 11387), S. yanoikuyae JCM 7371 (= GIFU 9882), and S. adhaesiva JCM 7370 (= GIFU 11458) are designated as the type strains of the three new species. Emended description of the type strain of S. capsulata is presented.


Stable cavitation at low ultrasonic intensities induces cell death and inhibits 3H-TdR incorporation by Con-A-stimulated murine lymphocytes in vitro.
Murine spleen cell suspensions stimulated by Concanavalin A (Con-A) were exposed to 1.6-MHz continuous-wave ultrasound at low intensities (spatial-peak values ranging from 16 to 300 mW/cm2) in the presence of a Nuclepore membrane that contained stabilized gas bodies. The ultrasonically activated gas bodies induced cell lysis and reduced the fraction of intact cells that excluded trypan blue. At a spatial-peak intensity of 75 mW/cm2 (spatial-average intensity 15 mW/cm2), Con-A-induced Methyl[3H]thymidine (3H-TdR) incorporation was reduced in cultures exposed at 24 or 48 hr after addition of Con-A but not at 7 or 13 hr. There was no observable effect on cell survival or 3H-TdR incorporation at spatial-peak intensities below 75 mW/cm2.


Structural characterization and functional analysis of juvenile hormone acid methyltransferase JHAMT3 from the silkworm, Bombyx mori.
Juvenile hormone acid methyltransferase (JHAMT) is a rate-limiting enzyme of juvenile hormone (JH) biosynthesis in insects. It transfers the methyl group of S-adenosyl methionine to either the carboxyl group of JH acids or farnesoic acid to produce JH. Six JHAMT paralogues have been identified in the silkworm (Bombyx mori); among them, JHAMT1 and JHAMT2 display a methyltransferase activity. Here, the three-dimensional crystal structure of inactive JHAMT3 and the binary complex of JHAMT3 with its cofactor S-adenosyl-l-homocysteine were determined through X-ray crystallization. Comparative structural analysis revealed that JHAMT3 adopted a similar structural pattern to that of functional JHAMT2, which comprised one core Rossmann fold domain and one substrate-binding domain. Similar to JHAMT2, JHAMT3 underwent a conformational change at the Rossmann fold domain because of cofactor binding, which promoted ligand accommodation. However, it exhibited a relatively rigid substrate-binding pocket compared with that of JHAMT2. JHAMT3 was also highly expressed in the silk gland of fourth- and fifth-instar B. mori larvae. The results of expression profiling combined with activity analysis suggested that JHAMT3 might function as a binding protein of JH acids for the regulation of JH acid titers. These findings provide a structural basis for enhancing the understanding of the physiological function of JHAMT3 and a rational framework for the development of potent and specific inhibitors of JHAMT family members.


Do we understand the prefrontal cortex?
Many suggestions have been made as to the functions of the prefrontal (PF) cortex. However, they involve labelling areas using psychological terminology. But what we need to know is how the PF cortex operates. We argue that understanding this must start with describing the flow of information. We illustrate this argument by considering three PF areas. Each has a unique pattern of inputs and outputs, and we suggest that the implication is that each performs a unique transformation from the inputs to the outputs. The caudal PF cortex transforms input that is maintained by attention or short-term memory into the target of the appropriate eye movement. The mid-dorsal PF cortex transforms input concerning the order of objects or actions into the target of the appropriate eye and hand movements, thus supporting sequences of action. The ventral PF cortex transforms input concerning an object or sound into prospective activity that encodes the associated object or sound. However, it is important to appreciate that the mid-dorsal and ventral PF cortex are specialized for encoding abstract transformations, irrespective of the specific actions or objects. The advantage is that this enables generalization to novel problems that have the same underlying logic. We account for the difference between fast learning and slow learning in this way. The human brain has co-opted these mechanisms so as to support intelligence. Non-verbal tests of IQ typically use sequences of letters, numbers or designs. These test the ability to understand the abstract rules that apply. Here the activations lie in the mid-dorsal PF cortex. Verbal tests typically assess the ability to understand semantic associations. These can be presented either in pictorial or verbal form. Here the activations lie in the ventral PF cortex.


Role of electron microscopy in the evaluation of soft tissue neoplasms, with emphasis on spindle cell and pleomorphic tumors.
The current significant role of transmission electron microscopy in the evaluation of soft tissue tumors when correlated with conventional histological and immunohistochemical studies is discussed for the following entities: myxofibrosarcoma, storiform-pleomorphic fibrosarcoma (malignant fibrous histiocytoma), and myofibrosarcoma; dermatofibrosarcoma protuberans; hemangiopericytoma; monophasic synovial sarcoma; extrarenal rhabdoid tumor; soft tissue perineurioma; and gastrointestinal stromal tumors, notably the so-called autonomic nerve variant.


Ocular Side Effects of Aromatase Inhibitor Endocrine Therapy in Breast Cancer - A Review.
Aromatase inhibitor therapy is currently the preferred choice in postmenopausal women with estrogen receptor positive breast cancer. This article reviews the ocular side effects of treatment with aromatase inhibitors (AIs) in patients with breast cancer. A comprehensive search was performed on PubMed, Web of Science and Google scholar. After duplication removal, 14 clinical studies and 5 case reports, published between 2008 and 2021, were identified. Most frequently, AI treatment resulted in minor to moderate dry eye symptoms. "De novo" onset of Sjogren syndrome during AI therapy was also reported. Retinal and optic nerve side effects varied from mild, subclinical anatomic and functional impairment to severe decreased vision, secondary to hemi-central retinal artery occlusion, bilateral optic neuritis or uveitis with bilateral macular edema. Visual disturbances encountered during AI treatment may be underestimated. Ophthalmic screening is important for early detection and appropriate treatment.


Capsid-targeted viral inactivation can destroy dengue 2 virus from within in vitro.
Capsid-targeted viral inactivation (CTVI) has emerged as a conceptually powerful antiviral strategy that exploits viral structural proteins to target a destructive enzyme specifically into progeny virions. We have recently demonstrated the principle of CTVI against dengue virus infection and observed a modest therapeutic effect in vitro (Arch Virol 2005, 150: 659-669). Here we tested a prophylactic model of CTVI, in which mammalian cells stably expressing the dengue 2 virus capsid protein fused to a nuclease were infected with dengue virus and determined the effects on progeny virion infectivity. CTVI efficiently destroyed dengue 2 virus from within and decreased the infectious titers by 10(3)- to 10(4)-fold, suggesting that CTVI has potential in the prophylactic application for dengue virus infection.


Dietary Flavone Baicalein Combinate with Genipin Attenuates Inflammation Stimulated by Lipopolysaccharide in RAW264.7 Cells or Pseudomonas aeruginosa in Mice via Regulating the Expression and Phosphorylation of AKT.
Mounting evidence has shown that single-targeted therapy might be inadequate to achieve satisfactory effects. Thus, drug combinations are gaining attention as they can regulate multiple targets to obtain more beneficial effects. Heat shock protein 90 (HSP90) is a molecular chaperone that assists the protein assembly and folding of client proteins and maintains their stability. Interfering with the interaction between HSP90 and its client proteins by inhibiting the latter's activity may offer a new approach toward combination therapy. The HSP90 client protein AKT plays an important role in the inflammatory response syndrome caused by infections. In this study, the dietary flavone baicalein was identified as a novel inhibitor of HSP90 that targeted the N-terminal ATP binding pocket of HSP90 and hindered the chaperone cycle, resulting in AKT degradation. Combining baicalein with genipin, which was extracted from Gardenia jasminoides, could inhibit the pleckstrin homology domain of AKT, significantly increasing the anti-inflammatory effects both in vitro and in vivo. This synergistic effect was attributed to the reduction in AKT expression and phosphorylation. Thus, elucidating the mechanism underlying this effect will provide a new avenue for the clinical application and development of synergistic anti-inflammatory drugs.


Nature and Quantum-Inspired Procedures - A Short Literature Review.
Even before quantum computers were known to the public, quantum-inspired algorithms were already introduced and acquired a well-deserved reputation. Their strength lies on their ability to combine innovative characteristics and superior results. In the almost 20 years that the field has existed, a huge number of results have been obtained. This chapter is an attempt to give a succinct but comprehensive review of the current state of affairs in the field of quantum-inspired-based metaheuristics, particularly geared towards those that can be used in existing quantum computers.


Revision of metal-on-metal hip arthroplasty in a tertiary center: a prospective study of 39 hips with between 1 and 4 years of follow-up.
Operative findings during revision of metal-on-metal hip arthroplasty (MOMHA) vary widely and can involve massive soft tissue and bone disruption. As a result, planning of theater time and resources is difficult, surgery is challenging, and outcomes are often poor. We describe our experience with revision of MOMHA and provide recommendations for management. We present the findings and outcomes of 39 consecutive MOMHAs (in 35 patients) revised in a tertiary unit (median follow-up time 30 (12-54) months). The patients underwent a preoperative work-up including CT, metal artifact reduction sequence (MARS) MRI, and blood metal ion levels. We determined 5 categories of failure. 8 of 39 hips had conventional failure mechanisms including infection and impingement. Of the other 31 hips, 14 showed synovitis without significant disruption of soft tissue; 6 had a cystic pseudotumor with significant soft tissue disruption; 7 had significant osteolysis; and 4 had a solid pseudotumor. Each category of failure had specific surgical hazards that could be addressed preoperatively. There were 2 reoperations and 1 patient (2 hips) died of an unrelated cause. Median Oxford hip score (OHS) was 37 (9-48); median change (ΔOHS) was 17 (-10 to 41) points. ΔOHS was similar in all groups-except those patients with solid pseudotumors and those revised to metal-on-metal bearings, who fared worse. Planning in revision MOMHA is aided by knowledge of the different categories of failure to enable choice of appropriate personnel, theater time, and equipment. With this knowledge, satisfactory outcomes can be achieved in revision of metal-on-metal hip arthroplasty.


Axin directs the amplification and differentiation of intermediate progenitors in the developing cerebral cortex.
The expansion of the mammalian cerebral cortex is safeguarded by a concerted balance between amplification and neuronal differentiation of intermediate progenitors (IPs). Nonetheless, the molecular controls governing these processes remain unclear. We found that the scaffold protein Axin is a critical regulator that determines the IP population size and ultimately the number of neurons during neurogenesis in the developing cerebral cortex. The increase of the IP pool is mediated by the interaction between Axin and GSK-3 in the cytoplasmic compartments of the progenitors. Importantly, as development proceeds, Axin becomes enriched in the nucleus to trigger neuronal differentiation via β-catenin activation. The nuclear localization of Axin and hence the switch of IPs from proliferative to differentiative status are strictly controlled by the Cdk5-dependent phosphorylation of Axin at Thr485. Our results demonstrate an important Axin-dependent regulatory mechanism in neurogenesis, providing potential insights into the evolutionary expansion of the cerebral cortex.


Modifiable factors associated with improved cardiac arrest survival in a multicenter basic life support/defibrillation system: OPALS Study Phase I results. Ontario Prehospital Advanced Life Support.
This study was conducted to identify modifiable factors associated with survival for prehospital cardiac arrest in a large, multicenter EMS system with basic life support/defibrillation (BLS-D) level of care. This observational cohort study constitutes Phase I of the 3-phase Ontario Prehospital Advanced Life Support (OPALS) Study. Included were all adults who had cardiac arrest before EMS arrival in 21 urban/suburban communities that operate under the jurisdiction of 1 ambulance services branch, have 911 telephone service, and provide ambulance defibrillation but no prehospital advanced life support (ALS). Central dispatch and ambulance records were reviewed according to the Utstein guidelines. Associations between multiple patient and EMS factors and survival to discharge were assessed by univariate then stepwise logistic regression analyses. From January 1, 1991, to January 31, 1995, 5,335 eligible patients were treated. Of these, 46.8% of cardiac arrests were witnessed by citizens, 14.5% received bystander CPR, 25.6% received CPR by fire or police, and 38.2% had an initial rhythm of ventricular fibrillation/ventricular tachycardia (VF/VT). The mean interval from call received to vehicle stopped was 6.7 minutes. Survival was 3.5% overall and 8.8% for VF/VT. Multivariate analysis found the following factors to be independently associated with survival (odds ratio with 95% confidence intervals): age.81 (. 73,.89), bystander-witnessed arrest 4.05 (2.78, 5.90), bystander CPR 2.98 (2.07, 4.29), CPR by fire or police 2.20 (1.46, 3.31), and response interval call received to vehicle stopped.76 (.71,.82). This represents the largest multicenter BLS-D study of prehospital cardiac arrest yet conducted and clearly indicates that patient survival may be improved by optimization of EMS response intervals, bystander CPR, as well as first-responder CPR by fire or police.


Mechanisms of exercise-induced pulmonary hypertension in patients with cardiac septal defects.
The objective of this study was to investigate mechanisms of exercise-induced pulmonary hypertension in patients with congenital cardiac septal defects. This was a randomized, placebo controlled, crossover drug trial in a single national pediatric cardiology centre that performs congenital cardiac defect surgery. There were 14 patients with cardiac septal defects and known exercise-induced pulmonary hypertension. The intervention consisted of 50 mg oral sildenafil versus placebo. Measurements included supine bicycle exercise echocardiography and oxygen uptake. The outcome measure was right-ventricular systolic pressure as estimated by Doppler tracings of tricuspid regurgitant jet as well as systolic and diastolic longitudinal myocardial velocities by color tissue Doppler echocardiography. Sildenafil did not change exercise right-ventricular systolic pressure during exercise; however, decreased systemic systolic pressure was seen. Enhanced biventricular longitudinal function with sildenafil compared with placebo was indicated by greater tissue Doppler velocities and displacement measurements during exercise. Finally, a less steep increase of right-ventricular pressure during exercise was associated with greater left-ventricular diastolic myocardial tissue Doppler velocity. Exercise-induced pulmonary hypertension in cardiac septal defects does not seem to have a pulmonary vasoconstrictive component, but it may be related to left-ventricular filling pressure. Furthermore, sildenafil improved biventricular systolic performance in this patient group, possibly related to decreased systemic afterload.


Functional assessment: Free thin anterolateral thigh flap versus free radial forearm reconstruction for hemiglossectomy defects.
To compare free thin anterolateral thigh (ALT) flap with free radial forearm (FRF) flap in the reconstruction of hemiglossectomy defects, and to introduce our methods and experience in the tongue reconstruction with free thin ALT flap. The clinicopathologic data of 46 tongue carcinoma cases hospitalized from December 2009 to April 2014 were obtained from Nangjing Stomatological Hospital, Medical School of Nanjing University. All the subjects were evaluated for the articulation and the swallowing function 3 months after the surgery. Among these 46 patients, 12 patients underwent tongue reconstruction after hemiglossectomy with ALT flap; 34 patients underwent tongue reconstruction with FRF flap. The differences in the incidence of vascular crisis, the speech and the swallowing function between two groups were not significant (P > 0.05). Thin ALT flap could be one of the ideal flaps for hemiglossectomy defect reconstruction with its versatility in design, long pedicle with a suitable vessel diameter, and the neglectable donor site morbidity.


Recovery of the failing canine heart with biventricular support in a previously fatal experimental model.
Prolonged normothermic ischemia in the canine model is generally fatal with standard resuscitative techniques. To determine whether such myocardial injury is recoverable with biventricular support, we subjected 10 dogs to 45 minutes of ischemia at 37 degrees C. After ischemia, the animals were supported for 24 hours with biventricular assist with the centrifugal pump. During early reperfusion, none of the hearts could sustain a stable rhythm or blood pressure. Myocardial adenosine triphosphate concentration, expressed as micromoles per gram of heart protein, was dramatically reduced from a control of 31.5 +/- 2.4 to 14.6 +/- 2.9 (p less than 0.01 versus control), a 54% reduction. Ultrastructural analysis did not reveal the explosive cell swelling of irreversible cell injury. After 12 hours of biventricular assist, developed pressure partially recovered to 60.0 +/- 10 mm Hg (p less than 0.01 versus control) and maximal positive dP/dt measured 2,649 +/- 412 mm Hg/sec (p less than 0.01 versus control). Adenosine triphosphate concentration increased to 25.2 +/- 5.5 (p less than 0.01 versus control). Electron microscopic examination showed less chromatin clumping, no further mitochondrial distortion, and more abundant glycogen. After 24 hours of biventricular assist, cardiac output in the seven dogs successfully weaned from biventricular assist measured 3.6 +/- 0.6 L/min, developed pressure recovered to 76.3 +/- 8.9 mm Hg, and its first derivative recovered to 4,282 +/- 585 mm Hg/sec (all measurements not significant compared with control). Examination by an electron microscope revealed no severe mitochondrial injury.


Transcription factor MdLSD1 negatively regulates α-farnesene biosynthesis in apple-fruit skin tissue.
α-Farnesene is a sesquiterpene present in plants. It was first discovered in apples. It plays an important role in the plant defence response and is considered a key factor in the occurrence of superficial scald. The gene encoding α-farnesene synthase, which is the last key enzyme in the biosynthetic pathway of α-farnesene in apple fruit, has become the primary target enzyme for controlling the genetic manipulation of α-farnesene biosynthesis. In this study, the yeast one-hybrid assay and the dual luciferase assay were used to ascertain the relationship between MdLSD1 and MdAFS. Real-time PCR was used to analyse the molecular mechanism underlying the regulation of MdAFS by MdLSD1. Our results revealed that transcription factor MdLSD1, which is closely related to programmed cell death in apple fruit tissues, binds to MdAFS. Transient transformation of apple skin with vectors overexpressing MdLSD1 showed that the gene negatively regulates MdAFS. Overall, we suggest that MdLSD1 negatively regulates MdAFS. Our results are of great significance for future research on the transcriptional regulation of the α-farnesene synthase gene and provide a new direction for exploring the specific mechanism of programmed cell death involved in superficial-scald incidence.


Determination of some anti-human immunodeficiency virus nucleosides by capillary zone electrophoresis-tandem mass spectrometry.
We describe the analysis of two potent anti-human immunodeficiency virus reverse transcriptase (RT-HIV) inhibitors, zidovudine (AZT) and stavudine (d4T), among a pool of natural nucleosides (A, C, G, U, T) by capillary zone electrophoresis (CZE)-ionspray-tandem mass spectrometry (MS/MS) in positive ion mode. Several volatile formic acid-ammonia buffers having the same ionic strength (50 mM) but different pH values varying in the 9-11 pH range were prepared and tested to determine the best electrophoretic migration conditions. Quantitative CZE-MS/MS analysis was performed using selected reaction monitoring (SRM) mode. Finally, this CZE-MS/MS procedure opens the possibility for future determination of several nucleoside RT-HIV inhibitors in cell pool samples.


Integrative multi-omics analysis depicts the methylome and hydroxymethylome in recurrent bladder cancers and identifies biomarkers for predicting PD-L1 expression.
Urinary bladder cancer (UBC) is a common malignancy of the urinary tract; however, the mechanism underlying its high recurrence and responses to immunotherapy remains unclear, making clinical outcome predictions difficult. Epigenetic alterations, especially DNA methylation, play important roles in bladder cancer development and are increasingly being investigated as biomarkers for diagnostic or prognostic predictions. However, little is known about hydroxymethylation since previous studies based on bisulfite-sequencing approaches could not differentiate between 5mC and 5hmC signals, resulting in entangled methylation results. Tissue samples of bladder cancer patients who underwent laparoscopic radical cystectomy (LRC), partial cystectomy (PC), or transurethral resection of bladder tumor (TURBT) were collected. We utilized a multi-omics approach to analyze both primary and recurrent bladder cancer samples. By integrating various techniques including RNA sequencing, oxidative reduced-representation bisulfite sequencing (oxRRBS), reduced-representation bisulfite sequencing (RRBS), and whole exome sequencing, a comprehensive analysis of the genome, transcriptome, methylome, and hydroxymethylome landscape of these cancers was possible. By whole exome sequencing, we identified driver mutations involved in the development of UBC, including those in FGFR3, KDMTA, and KDMT2C. However, few of these driver mutations were associated with the down-regulation of programmed death-ligand 1 (PD-L1) or recurrence in UBC. By integrating RRBS and oxRRBS data, we identified fatty acid oxidation-related genes significantly enriched in 5hmC-associated transcription alterations in recurrent bladder cancers. We also observed a series of 5mC hypo differentially methylated regions (DMRs) in the gene body of NFATC1, which is highly involved in T-cell immune responses in bladder cancer samples with high expression of PD-L1. Since 5mC and 5hmC alternations are globally anti-correlated, RRBS-seq-based markers that combine the 5mC and 5hmC signals, attenuate cancer-related signals, and therefore, are not optimal as clinical biomarkers. By multi-omics profiling of UBC samples, we showed that epigenetic alternations are more involved compared to genetic mutations in the PD-L1 regulation and recurrence of UBC. As proof of principle, we demonstrated that the combined measurement of 5mC and 5hmC levels by the bisulfite-based method compromises the prediction accuracy of epigenetic biomarkers.


In vivoblue light illumination for optogenetic inhibition: effect on local temperature and excitability of the rat hippocampus.
Objective.The blue light-activated inhibitory opsin, stGtACR2, is gaining prominence as a neuromodulatory tool due its ability to shunt-inhibit neurons and is being frequently used inin vivoexperimentation. However, experiments involving stGtACR2 use longer durations of blue light pulses, which inadvertently heat up the local brain tissue and confound experimental results. Therefore, the heating effects of illumination parameters used forin vivooptogenetic inhibition must be evaluated.Approach.To assess blue light (473 nm)-induced heating of the brain, we used a computational model as well as direct temperature measurements using a fiber Bragg grating (FBG). The effects of different light power densities (LPDs) and pulse durations on evoked potentials (EP) recorded from dentate gyrus were assessed. For opsin-negative rats, LPDs between 127 and 636 mW mm-2and pulse durations between 20 and 5120 ms were tested while for stGtACR2 expressing rats, LPD of 127 mW mm-2and pulse durations between 20 and 640 ms were tested.Main results.Increasing LPDs and pulse durations logarithmically increased the peak temperature and significantly decreased the population spike (PS) amplitude and latencies of EPs. For a pulse duration of 5120 ms, the tissue temperature increased by 0.6 °C-3.4 °C. All tested LPDs decreased the PS amplitude in opsin-negative rats, but 127 mW mm-2had comparatively minimal effects and a significant effect of increasing light pulse duration was seen from 320 ms and beyond. This corresponded with an average temperature increase of 0.2 °C-1.1 °C at the recorded site. Compared to opsin-negative rats, illumination in stGtACR2-expressing rats resulted in much greater inhibition of EPs.Significance.Our study demonstrates that light-induced heating of the brain can be accurately measuredin vivousing FBG sensors. Such light-induced heating alone can affect neuronal excitability. Useful neuromodulation by the activation of stGtACR2 is still possible while minimizing thermal effects.


Behavior of immature and middle-aged mice as a function of dietary protein.
The differential effects of continued maintenance on a 12% vs. 24% protein diet were examined in immature and middle-aged male and female C57BL/6J mice. Exploratory behavior, body weight and fluid consumption of water, saccharin and quinine were monitored over 9 weeks of presentation of the special diets. No rehabilitation datawere obtained. The dietary manipulations differentially effected the various groups, but the pattern of change in behavior was a function of the age and sex of the subject and the specific measure employed.


Diisocyanate modifiable commercial filter paper with tunable hydrophobicity, enhanced wet tensile strength and antibacterial activity.
Paper made of cellulose has the advantages of substantial resource, good biocompatibility, large operation scale, and low production cost, however, it is usually hindered to replace plastic due to its inferior stability to water and poor mechanical strength. Herein, commercial filter paper (FP) was reacted with methylene diphenyl diisocyanate (MDI) to fabricate modified paper with tunable hydrophobicity and enhanced wet tensile strength. Due to the formation of urethane linkages in the reaction, both hydrophobicity and wet mechanical properties can be tuned and improved by varying the MDI cross-linking agent, exhibiting highest water contact angle of 137.0° and wet tensile strength of 4.8 MPa. In addition, the modified MDI-FPs exhibited excellent antibacterial performance against Escherichia coli compared with the original FP. Overall, this study provides a more simple approach to modify cellulose paper to produce versatile cellulose-based materials that can potentially replace the non-biodegradable plastics.


Closure after gastrostomy button.
A gastrostomy device is removed from the gastrostoma when no longer needed. The aim of the study was to test the hypothesis of whether it is possible for the surgeon to decide which stoma has to be closed with a gastroraphy and which to leave for a spontaneous closure within a reasonable period of time. Out of a cohort of 321 patients, who had been operated with a video-assisted gastrostomy, we included all the 48 patients having had their gastrostomy button removed. These patients were carefully followed and the closure of the gastrostoma was registered. According to the institutional routine we waited at least 3 months after the removal of the gastrostomy device before suggesting to the child's guardians an operative closure of the stoma. In 26 patients the stoma closed within 3 months, whereas in 22 patients a surgical gastroraphy was performed. We found no differences between the two groups regarding the patients' diagnoses, the duration of the gastrostoma use or patient's age at the time of removal of the gastrostomy device. This study rejected the hypothesis of predictability of the gastrostoma closure. Thus, we recommend a routine expectance after the removal of a gastrostomy device for at least 1 month. If no spontaneous closure occurs, then a gastroraphy should be performed.


Death, taxes, and the genetic code?
The genetic code is an enduring feature of biology: only rare circumstances result in changes to translation of the code, at least in nature. Researchers are devising methods to engineer ribosome-synthesized polypeptides containing novel and potentially useful amino acids.


(CT)n and (GT)n microsatellites: a new class of genetic markers for Salmo trutta L. (brown trout).
Thirteen (GT)n and four (CT)n microsatellite loci (n = 10 or more and n = 20 or more, respectively) have been isolated from a partial genomic library of brown trout and sequenced. On average, a (GT)n repeat sequence occurs approximately every 23 kb and a (CT)n repeat sequence every 76 kb in brown trout genome. Primers for DNA amplifications using the polymerase chain reaction (PCR) were synthesized for three single locus microsatellites. Mendelian inheritance of the observed polymorphisms was confirmed in full-sib families. Four brown trout populations (10 unrelated individuals per population) were screened for polymorphism with these three microsatellite loci. The total number of alleles detected in the four populations is five at one locus, six at the other two microsatellite loci and is three, on average, per population. Heterozygosities range from 0.18 to 0.74. The largest differences in allelic frequencies occurred between the Mediterranean and the Atlantic populations: this result is congruent with previous allozymic data. The gene-centromere distances of the three microsatellite markers were determined on gynogenetic lines: post-reduction rates range from 0.17 to 0.60. For all the three microsatellite loci, the primers designed from brown trout sequences can be used in another closely related species of salmonid, the rainbow trout (Oncorhynchus mykiss). This last aspect supports the view that microsatellite markers may have wide application in genetic studies in salmonid species and fishes in general.


Seawater nitrogen concentration and light independently alter performance, growth, and resource allocation in the bloom-forming seaweeds Ulva lactuca and Ulvaria obscura (Chlorophyta).
Ulva lactuca and Ulvaria obscura are seaweeds that form green tides on Salish Sea shores. They have similar macroscopic morphologies but differ in their biochemistries and physiological responses. To understand how they are affected by changes in environmental conditions, a factorial experiment was conducted in which algae were grown in artificial seawater with either low (10 μM) or high (160 μM) nitrate (NO3-) concentrations at high (29 mol photons·m-2.day-1) and low (4 mol photons·m-2.day-1) light levels. Light and NO3- affected algal responses, but always independently. After two weeks, U. lactuca grown in high light were larger, had lower maximum quantum yields (MQYs), and lower nitrogen (N), carbon (C), pigment, and dimethylsulfoniopropionate (DMSP) concentrations, respectively, relative to algae in low light. In contrast, U. obscura growth was unaffected by light. Like U. lactuca, U. obscura grown in high light had lower MQYs, and N, pigment, and DMSP concentrations. In high light, U. obscura also had 89% higher dopamine concentrations and a tendency to fragment. Both U. lactuca and U. obscura grown in 160 μM NO3- were larger, had higher MQYs, and higher N, pigment, and DMSP concentrations, respectively, than algae in 10 μM NO3-. Also, when U. obscura was grown in the 160 μM NO3- medium, it significantly increased its surface area/mass ratio. Although both species grew faster in high NO3-, high light only promoted the growth of Ulva, which may explain the dominance of Ulva in summer months. High light was physiologically stressful to both species and caused increases in photoprotective mechanisms, such as the production of dopamine, a melanin precursor, in Ulvaria, and DMSP lysis in Ulva to generate antioxidants. Growing in 10 μM NO3- produced responses that were consistent with nitrogen limitation and had greater impacts on Ulvaria than Ulva, suggesting that Ulvaria responds more strongly to eutrophication.


Early life predictors of obstructive sleep apnoea in young adults: Insights from a longitudinal community cohort (Raine study).
Early-life obstructive sleep apnoea (OSA) predictors are unavailable for young adults. This study identifies early-life factors predisposing young adults to OSA. This retrospective study included 923 young adults and their mothers from the Western Australian Pregnancy Raine Study Cohort. OSA at 22 years was determined from in-laboratory polysomnography. Logistic regression was used to identify maternal and neonatal factors associated with OSA in young adulthood. OSA was observed in 20.8% (192) participants. Maternal predictors of OSA included gestational diabetes mellitus (odds ratio (OR) 9.54, 95% confidence interval (CI) 1.7, 58.5, P = 0.011), preterm delivery (OR 3.18, 95%CI 1.1,10.5, P = 0.043), preeclampsia (OR 2.95, 95%CI 1.1,8.0, P = 0.034), premature rupture of membranes (OR 2.46, 95%CI 1.2, 5.2, P = 0.015), age ≥35 years (OR 2.28, 95%CI 1.2,4.4, P = 0.011), overweight and obesity (pregnancy BMI≥25 kg/m2) (OR 2.00, 95%CI 1.2,3.2, P = 0.004), pregnancy-induced hypertension (OR 1.89, 95%CI 1.1,3.2, P = 0.019), and Chinese ethnicity (OR 2.36,95%CI 1.01,5.5, P = 0.047). Neonatal predictors included male child (OR 2.10, 95%CI 1.5,3.0, P < 0.0001), presence of meconium-stained liquor during delivery (OR 1.60, 95%CI 1.0,2.5, P = 0.044) and admission to special care nursery (OR 1.51 95%CI 1.0,2.2, P = 0.040). Higher birth lengths reduced OSA odds by 7% for each centimetre (OR 0.93, 95%CI 0.87, 0.99, P = 0.033). A range of maternal and neonatal factors predict OSA in young adults, including those related to poor maternal metabolic health, high-risk pregnancy and stressful perinatal events. This information could assist in the early identification and management of at-risk individuals and indicates that better maternal health may reduce the likelihood of young adults developing OSA.


Low genetic diversity and strong immunogenicity within the apical membrane antigen-1 of plasmodium ovale spp. imported from africa to china.
Malaria is still an important challenge for global public health because of its extensive mortality and morbidity. Plasmodium ovale is mainly distributed in tropical regions of Africa and Asia. it includes two distinct ovale malaria species, which are P. ovale curtisi and P. ovale wallikeri. Apical membrane antigen-1 (AMA-1) is an asexual blood-stage protein which is essential for Plasmodium. Thus far, no study on gene polymorphism and immunogenicity of P. ovale AMA-1 (PoAMA-1) has been conducted. Amplified poama1 gene products from 14 P ovale curtisi samples and 12 P ovale wallikeri samples imported from Africa to Jiangsu Province, China were sequenced and their polymorphisms were analyzed. We expressed recombinant PoAMA-1 (rPoAMA-1, 53 kDa) proteins in an E. coli expression system and evaluated immune responses against the rPoAMA-1 in BALB/c mice. We identified a synonymous mutation in nucleotide position 333 of the pocama-1 gene and powama-1 did not reveal any variation. The humoral and cellular immune responses to rPoAMA-1 were evaluated using enzyme-linked immunosorbent assay (ELISA) and flow cytometry. rPoAMA-1-immunized mice produced specific antibodies as verified by immunoblotting. The rPoAMA-1 induced high antibody titers (1: 640,000), and had high avidity indexes (an average of 78.63% and 83.40%). The antibodies also recognized the native proteins, namely, crude antigen from blood stages. Cross-reactivity between rPocAMA-1 and rPowAMA-1 was observed. Moreover, rPoAMA-1 s induced interferon (IFN)-gamma-secreting cells in mice and increased lymphocyte proliferation response. Low genetic diversity was observed in poama-1 from the Jiangsu Province imported malaria cases, and further studies conclusively showed its strong immunogenicity. Significant cross-reactivity was found between rPocAMA-1 and rPowAMA-1, suggesting that a single PoAMA-1 antigen could be used to diagnose P. ovale curtisi or P. ovale wallikeri patient simultaneously. However, further evaluation needs to be carried out to validate the potential and limitations of PoAMA-1 as a candidate vaccine.


Specific oxidative stress parameters differently correlate with nailfold capillaroscopy changes and organ involvement in systemic sclerosis.
Oxidative stress is suggested to be involved in the pathogenesis of systemic sclerosis (SSc). The aim of the present study was to clarify such a hypothesis by determination of four different plasmatic parameters of oxidative stress, and to define its role in the microvascular damage, assessed by nailfold capillaroscopy (NC). Plasma samples of 18 patients with SSc were analyzed. The biomarkers measured were: total antioxidant capacity, hydroperoxides (ROOHs), and sulfhydryl (SH) and carbonyl (CO) groups. Each patient had a detailed clinical assessment and underwent an NC. The results showed significantly increased ROOHs in SSc patients compared to control group (5.02 +/- 0.24 vs 3.28 +/- 0.19 micromol/l; p < 0.05). Plasmatic levels of SH groups were significantly lower in SSc (0.466 +/- 0.08 mmol/l) compared to control group (0.542 +/- 0.04 mmol/l; p < 0.002). Plasma levels of ROOHs correlated with the capillaroscopy semiquantitative rating scale score (p < 0.05) and with the rating system for avascular areas (p < 0.03). The levels of CO groups inversely correlated with modified Rodnan's skin score (p < 0.039) and were lower in patients with pulmonary fibrosis (p < 0.045), while the levels of SH groups were lower in those presenting gastrointestinal involvement (p < 0.029). The obtained data indicate augmented free radical-mediated injury in SSc and also show correlations among oxidative abnormalities, some clinical findings, and signs of a more severe microvascular involvement. These results give more evidence to the connection between oxidative impairment and SSc.


Has the prevalence, clinical presentation and social functioning of schizophrenia changed over the last 25 years? Nithsdale schizophrenia survey revisited.
Major changes in the treatment of schizophrenia have taken place in recent years, including a shift from inpatient to community care, and the introduction of second-generation antipsychotics and psychosocial interventions. Recent evidence has questioned the superiority of these interventions over older treatments. We wished to ascertain the impact of changes in the treatment of schizophrenia in a geographically-defined area, focusing on clinical symptoms and psychosocial outcome. In 2006, we replicated the survey carried out in 1981, identifying the population of people with schizophrenia living in Nithsdale, South-West Scotland, measuring prevalence using the 'key informant' method. We assessed their clinical and social functioning, using similar scales, and compared measures across both time periods. We identified 205 people with schizophrenia, a point prevalence of 3.59/1000 general population. In 2006, while fewer people experienced negative symptoms, a greater number experienced positive symptoms. There were no significant differences in the prevalence of tardive dyskinesia or Parkinsonism, though tremor was less common in the 2006 population. In 2006, there were fewer people living with a spouse/partner or parents, and fewer were in open employment in comparison to the 1981 population. The overall level of social adjustment had not changed. The clinical manifestation of schizophrenia appears to have changed over the last 25 years, within this geographically-defined area, the prevalence of negative symptoms having decreased, and positive symptoms seem to have increased. The level of social functioning does not appear to have changed over the last 25 years.


Population pharmacokinetics of mycophenolic acid during the first week after renal transplantation.
To investigate the population pharmacokinetics of mycophenolic acid (MPA) in adult kidney transplant recipients during the crucial first week after transplantation. Data were collected from 117 patients. MPA plasma concentrations were determined at t=0, 1, 2, 3 and 4 h after mycophenolate mofetil dosing on days 3, 5 and 7. Population analysis was performed using NONMEM. Covariates screened were sex, age, body weight, serum creatinine, creatinine clearance, serum albumin, days of therapy, diabetes mellitus, organ source (live or cadaveric) and co-therapy (tacrolimus or cyclosporine). Final model validity was evaluated using 200 boot strapped samples from the original data. Bias and precision were determined through comparison of observed and predicted concentrations. Individual concentration-time profiles showed evidence of an absorption lag time and enterohepatic recirculation of MPA in some patients on some occasions. The best base model had bi-exponential elimination with a typical population (SE%) apparent clearance (CL/F) of 29 l/h (5%) and apparent volume of the central compartment of 65 l (7%). CL/F decreased significantly with increasing serum albumin (1.42 l/h reduction in total plasma CL/F with each 1 g/l increase in albumin) and was 27% greater in patients receiving cyclosporine than in those receiving tacrolimus. Evaluation of the final model showed close agreement between pairs of boot strapped and final model parameter estimates (all differences <7%). Predictions were non-biased (0.11 mg/l) but imprecise (2.8 mg/l). Population pharmacokinetic parameters for MPA were determined. These can be used to achieve specific target MPA concentrations or areas under the concentration-time curve.


Studies of vegans: the fatty acid composition of plasma choline phosphoglycerides, erythrocytes, adipose tissue, and breast milk, and some indicators of susceptibility to ischemic heart disease in vegans and omnivore controls.
The fatty acid composition of erythrocytes, of plasma choline phosphoglycerides, and of adipose tissue, serum cholesterol, triglyceride and vitamin B12 concentrations, weights, heights and skinfold thickness were determined on 22 vegans and 22 age and sex matched omnivore controls. The fatty acid composition of breast milk from four vegan and four omnivore control mothers, and of erythrocytes from three infants breast fed by vegan mothers and six infants breast fed by omnivore control mothers was determined. The proportions of linoleic acid and its long-chain derivatives were higher, the proportion of the long-chain derivatives of alpha-linolenic acid was lower, and the ratio of 22:5omega3/22:6omega3 was greater in the tissues of the vegans and infants breast-fed by vegans than in controls; the most marked differences were in the proportions of linoleic (18:2omega6) and docosahexenoic (22:6omega3) acids. Weights, skinfold thickness, serum vitamin B12, cholesterol and triglyceride concentrations were less in vegans than in controls. The difference in serum cholesterol concentration was most marked. It is concluded that a vegan-type diet may be the one of choice in the treatment of ischemic heart disease, angina pectoris, and certain hyperlipidemias.


Adrenergic reinnervation of the denervated rat urinary bladder.
In rats undergoing unilateral extirpation of the pelvic ganglion, the adrenergic innervation disappeared on the ipsilateral side of the urinary bladder. It had reappeared after 6--9 weeks.


Effect of manipulative variables on the accuracy of crown wax patterns.
An instrument was developed to carve wax patterns in a reproducible manner. The methods of wax application to the die and wax removal during carving were similar to routine laboratory procedures. The wax patterns were invested in stone with the dies on which they were prepared, and sectioned to allow the determination of their adaptation on sectioned samples. Shrinkage of wax patterns on dies was found to create a marginal gap at shoulders and bevels. Further shrinkage observed after removal from the die was attributed to relaxation of elastic stresses in the wax. Carving of bevels improved adaptation to the die for samples with large gaps at the shoulder. This improved adaptation can be attributed to plastic deformation in the wax bevel. Remodeling of pattern margins was found to improve adaptation to the die, especially if carried out after the pattern was removed and replaced on the die. Remodeling the margin should involve heating with a spatula of a narrow band of wax around the margin up to the axial die wall over the entire shoulder width.


Allelic replacement in Staphylococcus aureus with inducible counter-selection.
A method for rapid selection of allelic replacement mutations in the chromosome of Staphylococcus aureus is described. Plasmid pKOR1, an Escherichia coli/S. aureus shuttle vector, permits rapid cloning via lambda recombination and ccdB selection. Plasmid transformation of staphylococci and growth at 43 degrees C, a non-permissive condition for pKOR1 replication, selects for homologous recombination and pKOR1 integration into the bacterial chromosome. Anhydrotetracycline-mediated induction of pKOR1-encoded secY antisense transcripts via the Pxyl/tetO promoter, a condition that is not compatible with staphylococcal growth, selects for chromosomal excision and loss of plasmid. Using this strategy, allelic replacements in S. aureus rocA were generated at frequencies that obviated the need for antibiotic marker selection.


[Does antibiotherapy prevent unnecessary prostate biopsies in patients with high PSA values?].
We investigated if antibiotherapy has any role on total PSA (tPSA), free PSA (fPSA) and fPSA/tPSA ratio in patients with tPSA higher than 2.5ng/ml. We also analyzed if it has any relation with prostate cancer diagnosis rate. A total 108 patients older than 50 years of age with lower urinary system sypmtoms and tPSA >2.5ng/ml were included in this study. Antibiotherapy was given to all the cases for three weeks. After that, transrectal ultrasound-guided prostate biopsies were taken from all the patients. Before and after antibiotherapy, "The International Prostate Symptom Score" (IPSS) and "National Institutes of Health Chronic Prostatitis Symptom Index" (NIH-CPSI) questinories are performed and serum tPSA, fPSA and fPSA/tPSA values were obtained. TPSA, fPSA and fPSA/tPSA ratio alterations prior to and after antibiotherapy did not show any statistically significant difference (p>0.05). When prostate adenocarcinoma was excluded, an statistically significant decrease was found in IPSS and NIH-CPSI scores for all cases. Antibiotherapy given to patients with PSA levels higher than threshold value has not led to significant change in prostate needle biopsy decision. Prostate biopsy should be considered without trying antibiotherapy in patients with high PSA values if a suspicion of prostatitis does not exist.


"The best laid schemes...": an evaluation of the extension of midwifery training in Scotland.
A research project was undertaken to evaluate the extension of midwifery training in Scotland. The evaluative criterion used was the new midwives' employment decisions. The data on students' valuation of their training suggest that they regard their time as having been well spent and other data show that socialization into midwifery increased with the longer training. Confidence in midwifery skills did not change markedly in association with the extension of training. Certain benefits of the longer course anticipated among midwives, are shown by these data not to have happened.