Identification of a unique membrane-bound molecule on a hemopoietic stem cell line and on multipotent progenitor cells.
Hemopoietic stem cells are a distinct population of cells that can differentiate into multilineages of hemopoietic cells and have long-term repopulation capability. A few membrane-bound molecules have been found to be preferentially, but not uniquely, present on the surface of these primitive cells. We report here the identification of a unique 105-kDa glycoprotein on the surface of hemopoietic stem cell line BL3. This molecule, recognized by the absorbed antiserum, is not present on the surface of myeloid progenitors 32D and FDC-P1 cells, EL4 T cells, and NIH 3T3 fibroblasts. This antiserum can also be used to block the proliferation of BL3 cells even in the presence of mitogen-stimulated spleen cell conditioned medium, which is known to have a stimulating activity on BL3 cells. It can also inhibit development of in vitro, fetal liver cell-derived multilineage colonies, but not other types of colonies, and of in vivo bone marrow cell-derived colony-forming unit spleen foci. These data suggest that gp105 plays an important role in hemopoietic stem cell differentiation.


Structures of two major histocompatibility complex class I genes of the rainbow trout (Oncorhynchus mykiss).
Here we describe two rainbow trout major histocompatibility complex (MHC) class I genes characterized from lambda phage genomic clones prepared from a single fish. Clone GC71 contains all exons except a leader peptide-encoding exon. An open reading frame is maintained, and thus the gene MhcOnmy-U71 could be expressed in this individual. The class I gene found on clone GC41 lacks exons encoding the leader peptide and cytoplasmic domain. This gene, MhcOnmy-U41p, is a pseudogene due to a deletion in the alpha(2) domain-encoding exon causing premature termination. Both the Onmy-U71 and Onmy-U41p genes are distinguished by long introns between the exons encoding the alpha(1) and alpha(2) domains. Clone GC41 also contains the 3' exons of the LMP7/ PSMB8 gene encoding the gamma-interferon-induced proteosome subunit of rainbow trout.


Quantum fidelity and thermal phase transitions.
We study the quantum fidelity approach to characterize thermal phase transitions. Specifically, we focus on the mixed-state fidelity induced by a perturbation in temperature. We consider the behavior of fidelity in two types of second-order thermal phase transitions (based on the type of nonanaliticity of free energy), and we find that usual fidelity criteria for identifying critical points is more applicable to the case of lambda transitions (divergent second derivatives of free energy). Our study also reveals that for fixed perturbations, the sensitivity of fidelity at high temperatures (where thermal fluctuations wash out information about the transition) is reduced. From the connection to thermodynamical quantities we propose slight variations to the usual fidelity approach that allow us to overcome these limitations. In all cases we find that fidelity remains a good precriterion for testing thermal phase transitions, and we use it to analyze the nonzero temperature phase diagram of the Lipkin-Meshkov-Glick model.


Regulation of the expression of key genes involved in HDL metabolism by unsaturated fatty acids.
The cardioprotective effects of HDL have been largely attributed to their role in the reverse cholesterol transport pathway, whose efficiency is affected by many proteins involved in the formation and remodelling of HDL. The aim of the present study was to determine the effects, and possible mechanisms of action, of unsaturated fatty acids on the expression of genes involved in HDL metabolism in HepG2 cells. The mRNA concentration of target genes was assessed by real-time PCR. Protein concentrations were determined by Western blot or immunoassays. PPAR and liver X receptor (LXR) activities were assessed in transfection experiments. Compared with the SFA palmitic acid (PA), the PUFA arachidonic acid (AA), EPA and DHA significantly decreased apoA-I, ATP-binding cassette A1 (ABCA1), lecithin-cholesterol acyltransferase (LCAT) and phospholipid transfer protein mRNA levels. EPA and DHA significantly lowered the protein concentration of apoA-I and LCAT in the media, as well as the cellular ABCA1 protein content. In addition, DHA repressed the apoA-I promoter activity. AA lowered only the protein concentration of LCAT in the media. The activity of PPAR was increased by DHA, while the activity of LXR was lowered by both DHA and AA, relative to PA. The regulation of these transcription factors by PUFA may explain some of the PUFA effects on gene expression. The observed n-3 PUFA-mediated changes in gene expression are predicted to reduce the rate of HDL particle formation and maturation.


RNA therapeutics: beyond RNA interference and antisense oligonucleotides.
Here, we discuss three RNA-based therapeutic technologies exploiting various oligonucleotides that bind to RNA by base pairing in a sequence-specific manner yet have different mechanisms of action and effects. RNA interference and antisense oligonucleotides downregulate gene expression by inducing enzyme-dependent degradation of targeted mRNA. Steric-blocking oligonucleotides block the access of cellular machinery to pre-mRNA and mRNA without degrading the RNA. Through this mechanism, steric-blocking oligonucleotides can redirect alternative splicing, repair defective RNA, restore protein production or downregulate gene expression. Moreover, they can be extensively chemically modified to acquire more drug-like properties. The ability of RNA-blocking oligonucleotides to restore gene function makes them best suited for the treatment of genetic disorders. Positive results from clinical trials for the treatment of Duchenne muscular dystrophy show that this technology is close to achieving its clinical potential.


[Ethics and clinical drug trials].
During the last decade, biomedical ethics has known a rapid development. Research in man, including clinical drug trials, must now take into account ethical and legal requirements. The concept of the inviolability of the human person constitutes the basic tenet of biomedical ethics. Four main principles ensue: the principles of autonomy, of nonmaleficience, of beneficience and of justice. Some rules derive from them, for instance veracity, confidentiality, fidelity and respect of intimacy with regard to the subject of an experiment. From a practical standpoint, the design of a clinical trial must be scientifically sound, otherwise it cannot be ethical. An informed consent must be obtained, in written form, from the subject in a clinical trial and he must be aware of his right to withdraw from it at any moment. Moreover, the clinical trial must be monitored throughout its duration with respect to ethics. An ethics of the methodology of clinical trials must therefore exist. Ethics in research should be a way of thinking and behaving, inherent to the research training. The researcher should be keenly conscious of the ethical requirements involved in his work, if he wishes to avoid eventual external interventions.


Three-in-one blocks with ropivacaine: evaluation of sensory onset time and quality of sensory block.
The purpose of this prospective, randomized, double-blinded study was to evaluate the sensory onset time and the quality of sensory block of ropivacaine, a new long-acting local anesthetic, compared with bupivacaine, for 3-in-1 blocks. Fifty ASA physical status I-III patients undergoing hip surgery after trauma were randomly assigned to two study groups of 25 patients each. The two study groups received a 3-in-1 block with either 20 mL of ropivacaine 0.5% or 20 mL of bupivacaine 0.5%. Blocks in both groups were performed using a nerve stimulator. The sensory onset time and the quality of sensory block was assessed by pinprick test in the central sensory region of each of the three nerves and compared with the same stimulation in the contralateral leg. We used a scale from 100% (normal sensation) to 0% (no sensory sensation). We did not find significant differences in sensory onset times between the ropivacaine group and the bupivacaine group (30+/-11 vs 32+/-10 min). The quality of sensory blocks was also comparable between the study groups (19%+/-20% vs 21%+/-15%). We conclude that the sensory onset time and quality of sensory block during 3-in-1 blocks performed with ropivacaine are comparable to those with bupivacaine. Ropivacaine is described as being less potent than bupivacaine, making this local anesthetic promising for 3-in-1 blocks because of its reportedly lower incidence of cardiovascular and central nervous system complications. Ropivacaine 0.5% has a sensory onset time and quality of sensory block during 3-in-1 blocks similar to that of bupivacaine 0.5%. Ropivacaine is described as being less potent than bupivacaine, making it a promising local anesthetic for 3-in-1 blocks because of its reportedly lower cardiovascular and central nervous system toxicity.


Clinical profile, multimodal imaging, and treatment response in macular serpiginous choroiditis.
To describe the clinical profile, multimodal imaging, and treatment response in macular serpiginous choroiditis (MSC). Clinical records of 16 eyes (14 patients) with MSC presenting to a tertiary eye care institute between 2015 and 2019 were analyzed retrospectively. Mean age of 14 patients presenting with MSC was 33 ± 13 yrs with 64% males and 36% females. Mean visual acuity of the eyes with MSC at presentation was 0.43 ± 0.46 (logMAR) improving to 0.16 ± 0.28 (logMAR) at final visit. Thirteen eyes (81.3%) had active lesion at presentation. Mantoux test was positive in seven patients (50%) and QuantiFERON TB gold test positive in 10 patients (71%). HRCT chest showed latent tuberculosis in seven patients (50%). All patients underwent multimodal imaging. All patients received oral steroids as treatment therapy; 11 patients also received immunosuppressives, nine patients received additional anti-tubercular therapy (ATT). Mean duration of follow-up for the patients was 18 ± 10 months. A total of eight (50%) eyes had recurrence of lesions after an average duration of 14 ± 14 (3-36) months and were restarted on the treatment as per the requirement. At final follow-up, all eyes showed a good response to treatment and had healed lesions. Comparing the final BCVA to the initial BCVA, 38% (n = 6) showed improvement, 56% (n = 9) remained stable, and 6% (n = 1) eyes worsened at the final follow-up. Clinical profile and presentation of MSC is similar to that of CSC, and combination treatment with intravenous methyl prednisolone (IVMP), steroids, immunosuppressives, and ATT can salvage vision. A high suspicion of associated tuberculosis in endemic regions should be kept in mind.


Development and use of chromosome segment substitution lines as a genetic resource for crop improvement.
CSSLs are a complete library of introgression lines with chromosomal segments of usually a distant genotype in an adapted background and are valuable genetic resources for basic and applied research on improvement of complex traits. Chromosome segment substitution lines (CSSLs) are genetic stocks representing the complete genome of any genotype in the background of a cultivar as overlapping segments. Ideally, each CSSL has a single chromosome segment from the donor with a maximum recurrent parent genome recovered in the background. CSSL development program requires population-wide backcross breeding and genome-wide marker-assisted selection followed by selfing. Each line in a CSSL library has a specific marker-defined large donor segment. CSSLs are evaluated for any target phenotype to identify lines significantly different from the parental line. These CSSLs are then used to map quantitative trait loci (QTLs) or causal genes. CSSLs are valuable prebreeding tools for broadening the genetic base of existing cultivars and harnessing the genetic diversity from the wild- and distant-related species. These are resources for genetic map construction, mapping QTLs, genes or gene interactions and their functional analysis for crop improvement. In the last two decades, the utility of CSSLs in identification of novel genomic regions and QTL hot spots influencing a wide range of traits has been well demonstrated in food and commercial crops. This review presents an overview of how CSSLs are developed, their status in major crops and their use in genomic studies and gene discovery.


Stimulus- and amino acid-induced calcium and potassium changes in rat neocortex.
Local changes in extracellular concentration of calcium ions, [Ca2+]o, and potassium ions, [K+]o, elicited through ionophoretic applications of the excitatory amino acids glutamate (Glu) and aspartate (Asp) or through cortical surface stimulations were measured in the rat motor cortex using paired ion-selective microelectrodes. Glu or Asp applications produced dose-dependent decreases in [Ca2+]o from a base line of 1.25 mM to as low as 0.08 mM and increases in [K+]o from 2.8 mM to as high as 13.3 mM. These ionic changes were accompanied by negative DC potential shifts of up to 15 mV. [K+]o changes were practically constant in all cortical layers, whereas [Ca2+]o changes were variable with depth, showing two localized marked maximums. One was observed in the superficial layers, 150-300 microns below cortical surface and the second at 1,100-1,300 microns, in a region corresponding roughly to layer V. Application of tetrodotoxin (TTX) onto the cortical surface abolished stimulus-evoked ionic changes, whereas amino acid-induced [Ca2+]o changes remained practically unaltered, and [K+]o changes were reduced by 20% on average. Selective degeneration of the pyramidal tract neurons was induced by performing chronic lesions of the pyramidal tract at the bulbar level. In these conditions, laminar profiles of amino acid-induced [Ca2+]o changes were considerably altered in the corresponding motor cortex. The maximum Ca2+ changes were then reduced by two-thirds, and the [Ca2+]o signals were practically constant throughout the cortex. Local cobalt (Co2+) or manganese (Mn2+) applications could abolish amino acid-evoked [Ca2+]o changes, whereas most of the [K+]o changes persisted. gamma-Aminobutyric acid applications could decrease [Ca2+]o signals to a large extent, whereas [K+]o changes were diminished by only 15-25% on average. These results show that the putative neurotransmitters Glu and Asp induce significant changes in [Ca2+]o and [K+]o in the rat cerebral cortex and indicate the possible origin of the ion fluxes in terms of neuronal elements and ionic channels: in particular, they indicate that the major part of the amino acid-induced [Ca2+]o maximum decreases can be ascribed to Ca2+ movements through channels located very likely on the pyramidal tract neurons in the motor cortex. They also indicate that significant reductions of the extracellular space volume occur during excitatory amino acid applications.


Quantitative analysis of dysautonomia in patients with autonomic dysreflexia.
Autonomic dysreflexia (AD) is a life-threatening condition for individuals with cervical or high-thoracic spinal cord injury (SCI). The profile of autonomic dysfunction in AD using validated clinical autonomic tests has not been described so far, although it could be useful to identify SCI patients at greater risk of developing AD non-invasively. With this objective, 37 SCI patients (27% female) were recruited, and hemodynamic and cardiac parameters were continuously monitored to determine the presence of AD, defined as an increase of systolic blood pressure of 20 mmHg or higher after bladder filling with saline. Then, standard autonomic function testing was performed, including Deep Breathing, Valsalva Manoeuvre and Tilt Table Test. Finally, baroreflex sensitivity (BRS), and spectral analysis of heart rate and blood pressure variability were measured at rest. Catecholamines and vasopressin levels were also measured at supine and upright positions. The severity of SCI was assessed through clinical and radiological examinations. AD was observed in 73.3% of SCI patients, being 63.6% of them asymptomatic during the dysreflexive episode. AD patients displayed a drop in sympathetic outflow, as determined by decreased noradrenalin plasma levels, reduced sympathovagal balance and increased BRS. In line with decreased sympathetic activity, the incidence of neurogenic orthostatic hypotension was higher in AD patients. Our results provide novel evidence regarding the autonomic dysfunction in SCI patients with AD compared to non-AD patients, posing non-invasively measured autonomic parameters as a powerful clinical tool to predict AD in SCI patients.


Safety and Readmission in Pediatric Ambulatory Surgery in a Tertiary Hospital.
To assess the safety and outcome of pediatric ambulatory surgery by measuring the rate of complications and readmission and identifying common risk factors for complications and readmission. A cross-sectional study was conducted at King Abdullah Specialist Children's Hospital (KASCH), Riyadh, Kingdom of Saudi Arabia, using the BESTCare 2.0 Health Information System (SKHIC, Riyadh, Saudi Arabia). All cases admitted under the pediatric ambulatory surgery unit from June 2015 to May 2018 were included. We reviewed 462 medical charts and recorded the variables of age, sex, American Society of Anesthesiologists (ASA) classification, complications, and readmission within one month of the surgery. SPSS (IBM Corp., Armonk, NY, USA) was used for data analysis. Associations between exposure variables (e.g., age, duration of surgery) and the outcome variables (e.g., rate of readmission and complications) were measured using the Chi-square test for categorical variables, the T-test and analysis of variance (ANOVA) for numerical and categorical variables, and logistic regression for multiple variables to control confounding variables. Approximately, 3.5% of the pediatric ambulatory surgery cases required readmission, and 10.6% of the patients had complications with zero mortality. All the variables had no significant association either with the readmission or complications (p > 0.05), except for the duration of surgery in minutes which was associated with complications (OR 1.006, 95% CI, 1.000-1.012, P = 0.035). Among pediatric ambulatory surgery cases, the mortality rate is 0%, with low complications and readmission rates. Also, the longer the surgery, the higher the risk of complications.


One-step synthesis of hybrid nanocrystals with rational tuning of the morphology.
Metal-sulfide hybrid nanocrystals (HNCs) have been of great interest for their distinguished interfacial effect, which gives rise to unique catalytic properties. However, most of the reported metal-sulfide HNCs were synthesized via two-step approaches and few were fabricated based on the one-step strategies. Herein, we report a facile one-pot synthesis of CuPt-Cu2S, Pt-Cu2S HNCs, and CuPt nanocubes by simply changing the Pt precursor types. 1-Hexadecanethiol (HDT) was employed in this system to mediate the reduction of metal precursors and also as capping agent and sulfur source. Moreover, CuPd-Cu2S and Au-Cu2S HNCs were successfully prepared by using this one-step method. The catalytic properties of the obtained three nanocrystals were investigated in hydrogenation of cinnamaldehyde. Results show that CuPt-Cu2S HNCs exhibited the highest conversion rate and the highest selectivity toward hydrocinnamaldehyde while 3-phenyl-1-propanol was the only product over Pt-Cu2S HNCs.


Renal resistive indexes: variability in Doppler US measurement in a healthy population.
To determine the variability in resistive index (RI) in normal kidneys, possible causes of variability, and consequences of reporting a single value. Doppler ultrasound RI measurements were obtained in the upper, middle, and lower regions of 118 kidneys in 58 healthy subjects (aged 24-70 years; 35 women, 23 men) who subsequently underwent angiography. The effects of sampling a particular parenchymal region, vascular territory, or kidney were assessed. Kidney region, vascular territory, and right versus left kidney had no consistently significant effect (P < or = .05) on RI. Age had a statistically significant effect. RI readings were highly correlated with each other both within a subject and within a kidney. The probability that a single RI value would exceed 0.70 in a healthy 45-year-old subject was 6%; this decreased to 3% when three readings were averaged. The variability of RI measurements in a kidney suggests that a number of RI readings should be averaged before a single representative value is reported.


Radiocolloid scintigraphy in Felty's syndrome.
The Tc-99m sulfur colloid liver-spleen scintigrams of nine patients with Felty's syndrome (a triad of rheumatoid arthritis, splenomegaly, and neutropenia) were reviewed. The characteristic scintigraphic findings include (1) moderate or severe splenomegaly, (2) the reversal of liver-to-spleen uptake ratio despite normal liver function tests, and (3) virtually no visualization of the bone marrow uptake and no pulmonary radiocolloid sequestration. In a late stage of cirrhosis of the liver, moderate-to-severe splenomegaly with the reversal of liver-to-spleen uptake ratio almost always accompanies abnormal liver function, and sometimes there may be associated pulmonary radiocolloid uptake. It was therefore concluded that in a proper clinical setting, a radiocolloid scintigraph may differentiate between Felty's syndrome and cirrhosis of the liver in a late stage.


Formation of antisperm antibodies in women treated with fallopian tube sperm perfusion.
Fallopian tube sperm perfusion (FSP) is a combination of ovarian stimulation and intra-uterine insemination using a large volume (4 ml) of inseminate containing 10(7)-10(8) spermatozoa. The inseminate will flush the Fallopian tubes and some of it will end up in the pouch of Douglas. In the present study, we have investigated whether the FSP method will result in the formation of serum antisperm antibodies in the female. A total of 184 treatment cycles were given to 128 women. The indications for treatment were: unexplained infertility (n = 35), various infertility diagnoses (n = 28) and donor insemination (n = 65). Prior to treatment, 11 (8.6%) women had a positive tray-agglutination test (Friberg) and/or a positive immunobead test. After completing one to four treatment cycles, another six (4.7%) women had developed serum antisperm antibodies. The antibodies induced by the treatment were of isotype IgM and directed against the tail-tip of the spermatozoa. Two of the women, who prior to the treatment had antisperm antibodies, showed an increase in antibody titre during treatment. There was no statistically significant difference in the pregnancy rate between the women with antisperm antibodies and the women without. In our opinion, the small risk of developing antisperm antibodies is no contra-indication for treating infertile couples with FSP.


Carpal tunnel syndrome following burns.
The commonest cause of carpal tunnel syndrome (CTS) is a congenital predisposition - the carpal tunnel is simply narrower in some people than in others. The development of CTS due to various burn causes has never been reported. This study describes some demographic features of all reported CTS cases following different types of burn in patients admitted to our burns centre in the USA. A retrospective case study was carried out to identify CTS following different types of burns among the patients admitted between January 2001 and January 2006. A descriptive data analysis was carried out to observe CTS following hand burns. There were 36 CTS cases in 28 patients presenting various kinds of burns in the upper extremities. The mean age of the CTS patients was 52 years (SD, 10.51 yr) and 79% were male. Of these, 57% had sustained thermal burns, 32% electrical burns, and the remainder had scalds or chemical burns. More patients (56%) had second-degree burns than third-degree burns and all but one of the patients with electrical burns had second-degree burns. This observation demonstrates that there were a considerable number of CTS cases following thermal burn injury compared to previous reporting. This study also suggests the need of a prospective study to examine the association between burns in the upper extremities and the likelihood of their progression to CTS and whether any specific type of burn is more likely to result in CTS.


Exposure to indoor biomass fuel pollutants and asthma prevalence in Southeastern Kentucky: results from the Burden of Lung Disease (BOLD) study.
Asthma is a chronic inflammatory respiratory disease, characterized by episodic and reversible airflow obstruction and airway hyperresponsiveness and is influenced by both genetic and environmental factors. The Burden of Obstructive Lung Disease (BOLD) survey was used to determine the prevalence of self-reported asthma in a target population of 325,000 adults aged > or =40 in Southeastern Kentucky. Postbronchodilator spirometry was used to classify subjects based on lung function. Risk factors for asthma in this population, in particular indoor usage of biomass fuels, were evaluated. The overall study population was comprised of 508 individuals, with 15.5% reporting current asthma and 5.8% reporting former asthma. In this population, the following risk factors for asthma were identified: female sex, smoking, less than a high school education, increasing body mass index (BMI), and a history of cooking indoors with coal and wood. Cooking indoors with wood and coal for more than 6 months of one's life was shown to significantly increase the odds of reporting current asthma (odds ratio (OR) = 2.3, confidence interval (CI) 1.1, 5.0), whereas no effect was seen from a history of heating indoors with wood and coal (OR = 0.8, CI 0.4, 1.8). Current or former asthma was reported by 21.3% of the adult population. A history of using biomass fuels when cooking indoors significantly increased the risk of reporting current asthma in this population.


Predicting happiness among centenarians.
Happiness is believed to evolve from the comparison of current circumstances relative to past achievement. However, gerontological literature on happiness in extreme old age has been limited. The purpose of this study was to determine how perceptions of health, social provisions, and economics link past satisfaction with life to current feelings of happiness among persons living to 100 years of age and beyond. A total of 158 centenarians from the Georgia Centenarian Study were included to conduct the investigation. Items reflecting congruence and happiness from the Life Satisfaction Index were used to evaluate a model of happiness. Pathways between congruence, perceived economic security, subjective health, perceived social provisions, and happiness were analyzed using structural equation modeling. Congruence emerged as a key predictor of happiness. Furthermore, congruence predicted perceived economic security and subjective health, whereas perceived economic security had a strong influence on subjective health status. It appears that past satisfaction with life influences how centenarians frame subjective evaluations of health status and economic security. Furthermore, past satisfaction with life is directly associated with present happiness. This presents implications relative to understanding how perception of resources may enhance quality of life among persons who live exceptionally long lives.


Popliteus strain with concurrent deltoid ligament sprain in an elite soccer athlete: a case report.
Case Report (Differential diagnosis). Differential diagnosis of knee pathology after trauma may be difficult when diagnosing an isolated popliteus strain and concurrent medial deltoid ligament sprain. Upon a thorough search of the published literature, the authors found no reports delineating a popliteus strain in professional soccer in the United States. The joints most affected by injury in soccer players are the knee and ankle joints. The purpose of this case report is to describe the presentation of and difficulties encountered in diagnosing a popliteus strain in a Major League Soccer athlete. During an in-season away game, an outside defender was slide-tackled from behind when his right shank was caught in an externally rotated position underneath himself and the opposing player. The initial point of contact was made to the proximal third of the posterior right shank with an anteromedially directed force. The medial longitudinal arch of the foot was forced into a more midfoot pronated position and the subtalar joint was forced into eversion. The athlete was diagnosed with a moderate strain of the right popliteus muscle with a concurrent medial deltoid ligament sprain of the right ankle. This mechanism of injury, pain with passive knee flexion and internal rotation during McMurray's test, pain with Garrick's Test and magnetic resonance imaging (MRI) study confirmed the diagnosis. The athlete returned to full ninety-minute game participation after an intensive 15-day rehabilitation program. This case is unique because the injury manifested itself at multiple joints and specifically involved the popliteus muscle. The mechanism of injury can be associated with many other soft tissue injuries to the knee, and thus, may not lead the clinician initially to consider the diagnosis of a popliteus strain. Diagnosis of this entity may be difficult due to the possible shared attachment of the popliteus muscle to the lateral meniscus, and the lack of available testing methods to assess damage to the popliteus muscle. Level 5.


[Large paraesophageal hernia at risk of torsion in an 84-year-old man].
We describe the diagnosis of a large paraesophageal hernia that showed a risk of torsion in an 84-year-old man who had good health status and no clinical antecedents of interest until the previous night when he woke up and felt dyspnea, some pain located in the epigastrium and a fever spike. After a short interview with ambiguous and inconclusive answers, the main diagnosis was based on the data obtained from the physical examination, the electrocardiogram, the results of the emergency blood tests, and the hydroaerial level that appeared on the standing chest x-ray; acute myocardial infarction and pulmonary embolism were excluded. Once the patient was stabilized, esophagogastroscopy was requested and some hours later the patient underwent the remaining examinations: intestinal transit, opaque enema and computed tomography scan, which are described in the text. The results of these examinations form the basis of a generic discussion about this case and a literature review from point of view of geriatrics. Few cases of large diaphragmatic hernias in octogenarians have been reported in the literature. We discuss the contribution of the techniques used in the diagnosis of this entity.


Molecular predictors of clinical outcome in patients with head and neck squamous cell carcinoma.
Head and neck squamous cell carcinoma (HNSCC) involves the upper aerodigestive tract and can destroy the structure and function of organs involved in voice, speech, taste, smell and hearing, as well as vital structures necessary for survival. HNSCC has long been a treatment challenge because of the high rate of recurrences and of advanced disease at the time of diagnosis. Molecular identification of tissue biomarkers in diagnostic biopsy specimens may not only identify patients at risk for developing HNSCC but may also select patients that may benefit from more aggressive treatment modalities. Several biomarkers studied to date such as the proteins p53, cyclin D1, p16, Cox-2 enzyme, epidermal growth factor and vascular endothelial growth factor receptors, matrix metalloproteinases and the Fhit marker for genomic instability could be manipulated for the therapeutic benefit of these patients. This review presents the most updated information on molecular biomarkers with the greatest prognostic potential in HNSCC and discusses some factors that contribute to the controversy concerning their prognostic importance.


Aggregate nonparametric safety analysis of traffic zones.
Exploring the significant variables related to specific types of crashes is vitally important in the planning stage of a transportation network. This paper aims to identify and examine important variables associated with total crashes and severe crashes per traffic analysis zone (TAZ) in four counties of the state of Florida by applying nonparametric statistical techniques such as data mining and random forest. The intention of investigating these factors in such aggregate level analysis is to incorporate proactive safety measures in transportation planning. Total and severe crashes per TAZ were modeled to provide predictive decision trees. The variables which carried higher weight of importance for total crashes per TAZ were - total number of intersections per TAZ, airport trip productions, light truck productions, and total roadway segment length with 35 mph posted speed limit. The other significant variables identified for total crashes were total roadway length with 15 mph posted speed limit, total roadway length with 65 mph posted speed limit, and non-home based work productions. For severe crashes, total number of intersections per TAZ, light truck productions, total roadway length with 35 mph posted speed limit, and total roadway length with 65 mph posted speed limit were among the significant variables. These variables were further verified and supported by the random forest results.


Influence of extracellular matrix on the proliferation and differentiation of glioma cells in culture.
The influence of extracellular matrix on the proliferation and differentiation of glioma cells in vitro was investigated by culturing glioma cells in collagen gel and on collagen and laminin films. Rat C-6 glioma cells extended thin cytoplasmic processes, proliferated, and differentiated in collagen gel and on collagen film. The cells were stellate with multiple thin processes like the astrocyte in vivo. The intracellular content of cyclic adenosine-monophosphate in rat C-6 glioma cells on collagen film increased approximately four fold over the control level. In laminin film culture, rat C-6 glioma cells extended thin cytoplasmic processes with many knotty structures and proliferated. These findings confirm that extracellular matrix induces the differentiation of glioma cells.


Adrenal incidentalomas and subclinical Cushing's syndrome: diagnosis and treatment.
Adrenal incidentaloma has become a frequent clinical dilemma. Even in the absence of specific clinical features of Cushing's syndrome, adrenocortical incidentalomas may display steroid secretory activity at different degrees. The recognition of endocrine and metabolic dysfunctions associated with subclinical hypercortisolism leads to current awareness about its potential consequences. Different protocols and threshold values to define normal cortisol secretion and diagnosis of subclinical Cushing's syndrome have been proposed, including recent practice guidelines for the diagnosis of overt Cushing's syndrome. Follow-up studies have provided additional data about the natural course of the disease and related cardiovascular and metabolic consequences. The study of bilateral adrenocorticotropin-independent macronodular adrenocortical hyperplasia in some familial cases offers a new approach to understanding the spectrum of subclinical cortisol hypersecretion. The prevalence of subclinical hypercortisolism may be higher than previously reported as more sensitive diagnostic criteria are now recommended. The absence of a single gold standard test, the diversity of diagnostic criteria and the requirement of subsequent meticulous biochemical evaluations before a decision for treatment represent a challenge for the clinical management of this condition.


Emergency Braking Intention Detect System Based on K-Order Propagation Number Algorithm: A Network Perspective.
In order to avoid erroneous braking responses when vehicle drivers are faced with a stressful setting, a K-order propagation number algorithm-Feature selection-Classification System (KFCS) is developed in this paper to detect emergency braking intentions in simulated driving scenarios using electroencephalography (EEG) signals. Two approaches are employed in KFCS to extract EEG features and to improve classification performance: the K-Order Propagation Number Algorithm is the former, calculating the node importance from the perspective of brain networks as a novel approach; the latter uses a set of feature extraction algorithms to adjust the thresholds. Working with the data collected from seven subjects, the highest classification accuracy of a single trial can reach over 90%, with an overall accuracy of 83%. Furthermore, this paper attempts to investigate the mechanisms of brain activeness under two scenarios by using a topography technique at the sensor-data level. The results suggest that the active regions at two states is different, which leaves further exploration for future investigations.


[Case study of vulvar malignant melanoma conducted at the Cinquantenaire General Hospital in Lubumbashi].
Primary malignant melanoma of the female genital tract is an extremely rare tumor. It most commonly affects the vulva then the cervix and the vagina. Vulvar cancer accounts for about 1% (all sites) of melanomas. In order of frequency, it occurs in the vagina, uterus and ovary. Less than 200 cases of vulvar cancers have been described worldwide. We here report a clinical case of malignant vulvar melanoma in a 72-year-old postmenopausal woman. Partial hemivulvectomy and bilateral superficial inguinal lymphadenectomy were performed. The postoperative course was favorable and the patient was discharged on postoperative day 15.


Effect of glucose variability on pathways associated with glucotoxicity in diabetes: Evaluation of a novel in vitro experimental approach.
Glycaemic variability (GV) has been hypothesized to increase the risk of diabetes complications; however, results of clinical studies are contradictory. The effect of GV on cell phenotypes has been investigated in vitro showing that GV may have more deleterious effect on cells that high glucose itself. However, methodology used to study GV in vitro differs significantly between studies and does not reflect in vivo situation. Therefore we aimed to establish clinically relevant an in vitro experimental approach for the study of GV that reflects intra-day glucose fluctuations of subjects with type 1 diabetes mellitus (T1DM) and of healthy subjects and to test how low and high GV affect expression of genes that protects cells from hyperglycaemia-induced damage. Human umbilical vein endothelial cells (HUVEC) were cultured 24h in medium with different glucose profiles: high GV, low GV and GV of healthy subjects-profiles created according to CGM of T1DM patients and healthy subjects. These profiles were compared to commonly used 5.5 and 25mmol/l glucose concentrations. Gene expression was determined using quantitative PCR. Our results showed general down-regulation of enzymes that are involved in the protection against hyperglycaemia-induced intracellular changes in both low and high GV compared to normal glycaemia similarly to the decrease induced by continuous hyperglycaemia. Gene expressions did not differ between high and low GV. Our data indicate that GV may have similar or even greater effect than continuous hyperglycaemia on the expression of several genes relevant to pathogenesis of diabetes microvascular complications.


SEPALLATA--like genes of Isatis indigotica can affect the architecture of the inflorescences and the development of the floral organs.
The architecture of inflorescence and the development of floral organs can influence the yield of seeds and have a significant impact on plant propagation. E-class floral homeotic MADS-box genes exhibit important roles in regulation of floral transition and differentiation of floral organs. Woad (Isatis indigotica) possesses unique inflorescence, floral organs and fruit. However, very little research has been carried out to determine the function of MADS-box genes in this medicinal cruciferous plant species. SEPALLATA orthologs in I. indigotica were cloned by degenerate PCR. The sequence possessing the highest identity with SEP2 and SEP4 of Arabidopsis were named as IiSEP2 and IiSEP4, respectively. Constitutive expression of IiSEP2 in Columbia (Col-0) ecotype of Arabidopsis led to early flowering, and the number of the flowers and the lateral branches was reduced, indicating an alteration in architecture of the inflorescences. Moreover, the number of the floral organs was declined, the sepals were turned into carpelloid tissues bearing stigmatic papillae and ovules, and secondary flower could be produced in apetalous terminal flowers. In 35S::IiSEP4-GFP transgenic Arabidopsis plants in Landsberg erecta (Ler) genetic background, the number of the floral organs was decreased, sepals were converted into curly carpelloid structures, accompanied by generation of ovules. Simultaneously, the size of petals, stamens and siliques was diminished. In 35S::IiSEP4-GFP transgenic plants of apetalous ap1 cal double mutant in Ler genetic background, the cauliflower phenotype was attenuated significantly, and the petal formation could be rescued. Occasionally, chimeric organs composed of petaloid and sepaloid tissues, or petaloid and stamineous tissues, were produced in IiSEP4 transgenic plants of apl cal double mutant. It suggested that overexpression of IiSEP4 could restore the capacity in petal differentiation. Silencing of IiSEP4 by Virus-Induced Gene Silencing (VIGS) can delay the flowering time, and reduce the number and size of the floral organs in woad flowers. All the results showed that SEPALLATA-like genes could influence the architecture of the inflorescence and the determinacy of the floral meristems, and was also related to development of the floral organs.


Calcium Silicate Improved Bioactivity and Mechanical Properties of Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) Scaffolds.
The poor bioactivity and mechanical properties have restricted its biomedical application, although poly(3-hydroxybutyrate-co-3-hydroxyvalerate) (PHBV) had good biocompatibility and biodegradability. In this study, calcium silicate (CS) was incorporated into PHBV for improving its bioactivity and mechanical properties, and the porous PHBV/CS composite scaffolds were fabricated via selective laser sintering (SLS). Simulated body fluid (SBF) immersion tests indicated the composite scaffolds had good apatite-forming ability, which could be mainly attributed to the electrostatic attraction of negatively charged silanol groups derived from CS degradation to positively charged calcium ions in SBF. Moreover, the compressive properties of the composite scaffolds increased at first, and then decreased with increasing the CS content, which was ascribed to the fact that CS of a proper content could homogeneously disperse in PHBV matrix, while excessive CS would form continuous phase. The compressive strength and modulus of composite scaffolds with optimal CS content of 10 wt % were 3.55 MPa and 36.54 MPa, respectively, which were increased by 41.43% and 28.61%, respectively, as compared with PHBV scaffolds. Additionally, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay indicated MG63 cells had a higher proliferation rate on PHBV/CS composite scaffolds than that on PHBV. Alkaline phosphatase (ALP) staining assay demonstrated the incorporation of CS significantly promoted osteogenic differentiation of MG63 cells on the scaffolds. These results suggest that the PHBV/CS composite scaffolds have the potential in serving as a substitute in bone tissue engineering.


Identification of Driver Genes and Interaction Networks Related to Brain Metastasis in Breast Cancer Patients.
Brain metastasis is a common complication of breast cancer (BC); however, the interaction networks and driver genes that lead to brain metastasis in BC patients are still unknown. In this study, we employed bioinformatics analyses to discover hub genes and long noncoding RNA- (lncRNA-) protein-coding gene (PCG) networks related to BC brain metastasis (BCBM). Firstly, we screened differentially expressed PCGs and lncRNAs in normal and BCBM samples using the GSE52604 dataset. Subsequently, differentially expressed genes (DEGs) and overall interaction networks were constructed, and topological degrees were analyzed to identify potential driver genes. After identifying the hub pathogenic module by weighted gene coexpression network analysis (WGCNA), the genes in the hub module were evaluated for functional enrichment. Finally, we constructed multiple interaction networks associated with BCBM and identified seven potential driver genes, out of which MYBPC1 was the only overlapping gene in the adopted analytical methods. It is worth mentioning that we validated the prognostic value of the identified hub genes in TCGA database and evaluated the prediction ability of MYBPC1 in the GSE38057 dataset. In addition, the CIBERSORT algorithm revealed changes in the immune microenvironment. In conclusion, the driver PCGs and lncRNAs in the interaction networks can be utilized as a promising therapeutic strategy for the treatment of brain metastasis in BC patients.


Influence of light finger touch on postural stability during upright stance with cold-induced plantar hypoesthesia.
The present study investigated the influence of light finger touch on postural stability during an upright stance with or without cold-induced plantar hypoesthesia. Participants were 21 healthy right-handed men (20-33 years), randomly assigned to a normal temperature group (NML-group; n = 11) or cold-induced plantar hypoesthesia group (COLD-group; n = 10). Participants performed two standing tasks with their eyes closed: 1) stand upright with the feet comfortably apart (normal standing); and 2) with the feet together in a side-by-side stance (Romberg standing). The order of the tasks was randomized across participants. After a rest period, they were asked to perform each standing task and let their right index finger lightly touch the fixed surface. Before the each task, participants in the COLD group were conducted a plantar intermediate hypothermic exposure. Postural sway was decreased by providing additional haptic sensory input through the light touching regardless of standing task difficulty with a normal plantar temperature. Although postural sway under plantar hypoesthesia is decreased by light touching, the effect was greater during the difficult posture task. These results suggest that, when the sensory information from the plantar surface is decreased, additional haptic input from the hand or finger might provide supplementary information about the relative changes in one's own body orientation. Therefore, sensory information provided by touching became relatively more important to maintaining postural control during the relatively difficult postural task performed under the reduced plantar sensitivity condition.


Use of melatonin: a single-centre audit.
Sleep disorders in childhood are common. Melatonin is prescribed by UK community paediatricians to treat sleep disorders, but practice is not standardised. This audit reviewed melatonin prescribing within a community paediatric department in a 12-month period. 682 children received melatonin prescriptions; a random sample of 198 records were reviewed. The most common underlying condition was autism spectrum disorder (ASD) in 28%. 41% had no underlying diagnosis when melatonin was initiated and were waiting for neurodevelopmental/ASD assessment. 42% were on melatonin for at least 2 years. Further work is required to optimise melatonin prescribing practice for children and young people.


Three Types of Elicitors Induce Grapevine Resistance against Downy Mildew via Common and Specific Immune Responses.
Three recognized plant defense stimulators (PDS), methyl jasmonate (MeJA), benzothiadiazole (BTH) and phosphonates (PHOS), were sprayed on grapevine Vitis vinifera cuttings and conferred resistance to the biotrophic pathogen Plasmopara viticola. The effects on molecular defense-related genes and polyphenol content (stilbenes and flavanols) were revealed at 6 and 8 days post-elicitation. The transcript accumulation was consistent with the signaling pathway specific to the elicitor, salicylic acid for BTH, and jasmonic acid for MeJA, with some cross-talks. PHOS tended to modulate the defense responses like BTH. Moreover, in response to a downy mildew inoculation, the leaves pre-treated with PHOS and BTH overproduced pterostilbene, and after MeJA treatment, piceids and ε-viniferin, compared to uninoculated elicitor-treated leaves. These results provide evidence of the different modes of action of PDS and their role in sustainable viticulture.


Neuroimaging examination of newborns in vertically acquired infections.
Congenital/perinatal nervous system (CNS) infections are an important cause of mortality and morbidity in neonatal period, and long-term sequelae. Many pathogens can lead to infections frequently involving the CNS, with possible disruption of brain development, which often is related to gestational age of maternal infection. The mechanism of infection and damage is different among the infectious agents, leading to more specific pathologic findings. It is necessary in newborns with confirmed or suspected CNS infection to undergo investigation by neuroimaging techniques to help healthcare providers, give adequate treatment and follow-up care and counsel parents. Computed tomography, Magnetic Resonance Imaging and cerebral ultrasonography are fundamental tools in evaluating infants with suspected or proved congenital/perinatal infections. Each imaging technique has its advantages, disadvantages and limits, since they are sometimes complementary.


Expression pattern, ethanol-metabolizing activities, and cellular localization of alcohol and aldehyde dehydrogenases in human pancreas: implications for pathogenesis of alcohol-induced pancreatic injury.
Alcohol dehydrogenase (ADH) and aldehyde dehydrogenase (ALDH) are major enzymes responsible for metabolism of ethanol. Genetic polymorphisms of ADH1B, ADH1C, and ALDH2 occur among racial populations. The metabolic effect and metabolites contribute to pathogenesis of pancreatic injury. The goal of this study was to determine the functional expressions and cellular localization of ADH and ALDH families in human pancreas. Fifty five surgical specimens of normal pancreas as well as 15 samples each for chronic pancreatitis and pancreatic cancer from archival formalin-fixed paraffin-embedded tissue specimens were investigated. Class-specific antibodies were prepared by affinity chromatographies from rabbit antisera raised against recombinant human ADH1C1, ADH4, ADH5, ADH7, ALDH1A1, ALDH2, and ALDH3A1. The isozyme expression patterns of ADH/ALDH were identified by isoelectric focusing, and the activities were assayed spectrophotometrically. The protein contents of ADH/ALDH isozymes were determined by immunoblotting, and the cellular localizations were detected by immunohistochemistry and histochemistry. At 33 mM ethanol, pH 7.5, the activities were significantly different between allelic phenotypes of ADH1B. The activity of ALDH2-inactive phenotypes was slightly lower than ALDH2-active phenotypes at 200 microM acetaldehyde. The protein contents were in the following decreasing order: ALDH1A1, ALDH2, ADH1, and ADH5. ADH1B was detected in the acinar cells and ADH1C in the ductular, islet, and stellate cells. The expression of ADH1C appeared to be increased in the activated pancreatic stellate cells in chronic pancreatitis and pancreatic cancer. Alcohol dehydrogenase and ALDH family members are differentially expressed in the various cell types of pancreas. ADH1C may play an important role in modulation of activation of pancreatic stellate cells.


Comparative Appraisal of Biomass Production, Remediation, and Bioenergy Generation Potential of Microalgae in Dairy Wastewater.
The present study is a trail to integrate the phycoremediation and bioenergy production from microalgal species cultivated in the dairy wastewater (DWW). Higher biomass productivities for Chlorella pyrenoidosa (24.44 ± 8.02 mg L-1d-1), Anabaena ambigua (23.64 ± 5.69 mg L-1d-1) and Scenedesmus abundans (18.72 ± 2.06 mg L-1d-1) were recorded in 3:1 DWW over the control. The microalgal species have effectively reduced the BOD by 56%, COD by 77%, nitrate by 88%, and phosphate by 85% following 25 days of the cultivation in the 3:1 DWW. The total lipid content was 10.36, 13.13, and 16.93% of dry matter of biomass in C. pyrenoidosa, A. ambigua, and S. abundans, respectively following 25 days of cultivation in the 3:1 ratio of DWW. The biochemical characterization revealed that the protein content was 21.8% in C. pyrenoidosa, 17.73% in A. ambigua and 34.06% in S. abundans. The estimation of theoretical methane potential suggested that the microalgal species have the desirable possibility of biogas generation. The results have marked the achievability of an integrated process for the remediation and bioenergy production by the employment of microalgal species.


Bioresource Nutrient Recycling in the Rice-Wheat Cropping System: Cornerstone of Organic Agriculture.
This study evaluated the impact of conventional practices (fertilizer alone) and diverse farming approaches (such as green manuring, farmyard manure application, rice-residue incorporation, residue mulching, residue removal and residue burning) on soil attributes. A total of thirty-five farm sites were selected, with five sites (replications) for each farming approach system, which were used over the past three years in the study farms. Characterization of rice residues of all cultivars, green manure crop (sesbenia: Sesbania sesban) and decomposed farmyard manure samples showed differential behaviours for macronutrients and micronutrients. Continuous application of inorganic fertilizers significantly influenced soil attributes, especially electrical conductivity, nutrient contents, bacterial and fungal population and soil enzymatic attributes. The crop residue treatments favourably influenced the soil parameters over the control. Crop residue incorporation or burning significantly increased soil available potassium, microbial biomass, enzymatic activities and organic carbon when compared with applications of chemical fertilizer alone, while total nitrogen content was increased by residue incorporation. However, green manuring and farmyard manure applications showed inferior responses compared with residue management treatment. It is therefore recommended that bioresources should be managed properly to warrant improvements in soil properties, nutrient recycling and the sustainability for crop productivity, in order to achieve sustainable development goals for climate action.


Chemoselective synthesis of sialic acid 1,7-lactones.
The chemoselective synthesis of the 1,7-lactones of N-acetylneuraminic acid, N-glycolylneuraminic acid, and 3-deoxy-d-glycero-d-galacto-nononic acid is accomplished in two steps: a simple treatment of the corresponding free sialic acid with benzyloxycarbonyl chloride and a successive hydrogenolysis of the formed 2-benzyloxycarbonyl 1,7-lactone. The instability of the 1,7-lactones to protic solvents has been also evidenced together with the rationalization of the mechanism of their formation under acylation conditions. The results permit to dispose of authentic 1,7-sialolactones to be used as reference standards and of a procedure useful for the preparation of their isotopologues to be used as inner standards in improved analytical procedures for the gas liquid chromatography-mass spectrometry (GLC-MS) analysis of 1,7-sialolactones in biological media.


Analysis of mitogen-activated protein kinase signaling specificity in response to hyperosmotic stress: use of an analog-sensitive HOG1 allele.
When confronted with a marked increase in external osmolarity, budding yeast (Saccharomyces cerevisiae) cells utilize a conserved mitogen-activated protein kinase (MAPK) signaling cascade (the high-osmolarity glycerol or HOG pathway) to elicit cellular responses necessary to permit continued growth. One input that stimulates the HOG pathway requires the integral membrane protein and putative osmosensor Sho1, which recruits and enables activation of the MAPK kinase kinase Ste11. In mutants that lack the downstream MAPK kinase (pbs2Delta) or the MAPK (hog1Delta) of the HOG pathway, Ste11 activated by hyperosmotic stress is able to inappropriately stimulate the pheromone response pathway. This loss of signaling specificity is known as cross talk. To determine whether it is the Hog1 polypeptide per se or its kinase activity that is necessary to prevent cross talk, we constructed a fully functional analog-sensitive allele of HOG1 to permit acute inhibition of this enzyme without other detectable perturbations of the cell. We found that the catalytic activity of Hog1 is required continuously to prevent cross talk between the HOG pathway and both the pheromone response and invasive growth pathways. Moreover, contrary to previous reports, we found that the kinase activity of Hog1 is necessary for its stress-induced nuclear import. Finally, our results demonstrate a role for active Hog1 in maintaining signaling specificity under conditions of persistently high external osmolarity.


Elevated serum levels of interleukin-5 in patients with the syndrome of episodic angioedema and eosinophilia.
The syndrome of episodic angioedema and eosinophilia is characterized by cyclic edema, marked peripheral blood eosinophilia, and eosinophil degranulation in the dermis. Using a sensitive immunoenzymetric method, we measured serum interleukin (IL)-5 levels in four patients with this syndrome. We also determined the percentage of activated T cells in the peripheral blood of a new patient before and during an attack. In the patient presented, IL-5 levels peaked several days before maximal eosinophilia and then declined. This patient's lymphocytes showed an increased percentage, 28% (normal 2% to 3%), of activated T cells staining for both CD3 and HLA-DR 10 days before maximal eosinophilia, but no increase at the time of peak eosinophilia. In serum from three previously reported cases, elevated serum IL-5 levels were found during attacks. After glucocorticoid administration, IL-5 levels became undetectable in three of the four patients. Production of IL-5 is likely an important determinant of the pathophysiology of this syndrome.


Cognitive impairment after tick-borne encephalitis.
Tick-borne encephalitis (TBE) is a potentially debilitating disease caused by the TBE virus. Natural foci of TBE were localized in the Lublin region (Poland). We observed the growing tendency of the incidence of TBE in the Lublin region at the beginning of the 1990s. Farmers and forestry workers were most often infected due to the occupational risk. Forty farmers with TBE since the beginning of the year 2001 were included in the study. Serological measurements as well as medical and neuropsychological examination confirmed the infection. Dementia and mild cognitive impairment were diagnosed on the basis of DSM-IV-R, ICD-10 and Petersen criteria, respectively. Long-term consequences of the illness have been reported in 22 subjects. In this group, cognitive impairment of the amnestic type was diagnosed. No decline in cognitive status was observed in the examined patients for 6 months. The diagnosis of human TBE is usually based on serology, which has a number of pitfalls. The outcome of the disease can mimic many neurological diseases. For that reason, the history of farmers with cognitive decline should be carefully examined. There is reasonable likelihood of cognitive decline in patients with TBE.


Effects of bile salt infusion on chlorpromazine-induced cholestasis in the isolated perfused rat liver.
The present study has demonstrated that tauroursodeoxycholate (TUDC), but not taurocholate, can reverse chlorpromazine (CPZ)-induced cholestasis in the isolated perfused rat liver. At an infusion rate of 1.5 mumol/min, TUDC led to restoration of bile flow in the perfused rat liver made cholestatic by the addition of 250 microM CPZ. This reversal was accompanied by an increased excretion of CPZ and its metabolites. A higher infusion rate of 5.0 mumols TUDC/min, however, led to only a transient increase in bile flow and to no increase in CPZ excretion. In contrast to the effects of TUDC, infusion of taurocholate led to an exacerbation of CPZ-induced cholestasis. The differences in the efficacy of the two bile salts may be due to their relative detergent (hydrophobic) properties.


Toxicity of industrial wastes and waste leaching test eluates containing organic compounds.
Leaching tests, CEN prEN 12457-2, CEN PrEN 12457-3, and NEN 7349, were conducted for varnish residue and urea resin waste, two industrial wastes containing organic chemicals. The leaching test eluates were analyzed for solvent concentrations and total organic carbon. Aqueous leaching tests were found to be suitable for both chemical and biological testing. Ecotoxicity was assessed by luminescent bacteria, plant root growth, reverse electron transport, and ToxiChromopad. The eluates were highly toxic but the toxicity decreased in later stages of the multistep leaching test (NEN 7349). Urea resin eluates were significantly less toxic than varnish residue eluates. The solid wastes markedly inhibited plant seed germination before and after the leaching test (CEN prEN 12457-3). The solid wastes were not biologically degradable in the standard test (ISO 14593), whereas the eluates from the CEN prEN 12457-3 test degraded slowly.


Novel missense mutation (Leu466Arg) of the DAX1 gene in a patient with X-linked congenital adrenal hypoplasia.
We identified a DAX1 missense mutation, a substitution of arginine for leucine at codon 466 (Leu466Arg), in an infant with X-linked congenital adrenal hypoplasia (AHC). A heterozygous substitution, Leu466Arg, was also identified in his mother and sister. Since leucine at position 466 is well conserved among other orphan nuclear hormone receptor superfamilies and Leu466Arg was not detected among 50 normal Japanese control individuals, the mutation is most likely responsible for X-linked AHC. It is interesting to note that Leu466Arg among all mutations ever reported is located at the most C-terminal region of the DAX-1 protein. Most mutations identified previously were located in the C-terminal presumptive ligand binding domain. Hence, the C-terminal end of the DAX-1 protein may play an important role in the biological function, such as in normal adrenal embryogenesis.


Surface electromyography as a tool to study the head rest comfort in cars.
In this study, two methods were presented in order to evaluate objectively the comfort of head rests in cars using surface electromyography (SEMG). First, the Amplitude Probability Density Function (APDF) was computed. Extremely low level intensity activities were measured. No significant differences were observed on SEMG global activity between experiments with and without a head rest. Second, instead of computing one APDF, several APDF were calculated over time. Analyses focused on the rate of evolution of APDF parameters. This was a time varying APDF (TAPDF). This last method allowed a better diagnosis than the first one. For drivers, significant decreases in SEMG activity were observed when using a head rest. For passengers, a contradictory increase in SEMG activity was observed during experiments with a head rest. This observation was explained by the seat back angle chosen by passengers.


The Autonomy Over Smoking Scale.
Our goal was to create an instrument that can be used to study how smokers lose autonomy over smoking and regain it after quitting. The Autonomy Over Smoking Scale was produced through a process involving item generation, focus-group evaluation, testing in adults to winnow items, field testing with adults and adolescents, and head-to-head comparisons with other measures. The final 12-item scale shows excellent reliability (alphas = .91-.97), with a one-factor solution explaining 59% of the variance in adults and 61%-74% of the variance in adolescents. Concurrent validity was supported by associations with age of smoking initiation, lifetime use, smoking frequency, daily cigarette consumption, history of failed cessation, Hooked on Nicotine Checklist scores, and Diagnostic and Statistical Manual of Mental Disorder (4th ed., text rev.; American Psychiatric Association, 2000) nicotine dependence criteria. Potentially useful features of this new instrument include (a) it assesses tobacco withdrawal, cue-induced craving, and psychological dependence on cigarettes; (b) it measures symptom intensity; and (c) it asks about current symptoms only, so it could be administered to quitting smokers to track the resolution of symptoms.


Forensic Radiology: An Update.
Forensic radiology is a specialized area of medical imaging using radiological techniques to assist physicians and pathologists in matters related to the law. The forensic application of diagnostic medical radiology can be applied in many fields; the prime target of evaluation is the osseous skeleton, but soft tissues and abdominal and thoracic viscera may offer key findings. The technological progress in clinical radiology provides a lot of potential tools to forensic radiology, allowing wider fields of applications in this matter.


T cell antigen receptor ubiquitination is a consequence of receptor-mediated tyrosine kinase activation.
Engagement of the T cell antigen receptor results in both its phosphorylation and its ubiquitination. T cell antigen receptor ubiquitination was evaluated in Jurkat, a well characterized human T leukemia cell line. Treatment of cells with the tyrosine kinase inhibitor herbimycin A resulted in an inhibition of receptor ubiquitination. Consistent with this, pervanadate, which increases cellular tyrosine phosphorylation, enhanced receptor ubiquitination. A requirement for receptor-mediated tyrosine kinase activity for ubiquitination was confirmed in cells lacking the tyrosine kinase p56lck and also in cells that are defective in expression of CD45, a tyrosine phosphatase that regulates the activity of p56lck. The need for tyrosine kinase activation for ubiquitination was not bypassed by directly activating protein kinase C and stimulating endocytosis of receptors. These observations establish ubiquitination of the T cell antigen receptor as a tyrosine kinase-dependent manifestation of transmembrane signaling and suggest a role for tyrosine phosphorylation in the ligand-dependent ubiquitination of mammalian transmembrane receptors.


Factors associated with self-reported pain and hand function following surgical A1 pulley release.
Surgical A1 pulley release can considerably reduce pain and improve hand function, but individual outcomes are highly variable. This study aimed to identify factors contributing to self-reported pain and hand function 3 months postoperatively. We included 2681 patients who had received surgical treatment for a trigger finger or thumb and who completed the Michigan Hand outcomes Questionnaire (MHQ). Hierarchical linear regression models were used to investigate patient and clinical characteristics associated with postoperative pain and hand function. For both pain and hand function, the most influential factors associated with worse outcomes were worse MHQ scores at baseline (β 0.38 and 0.33, respectively) and ≥3 preoperative steroid injections (β -0.36 and -0.35). These factors indicated that patients with severe preoperative symptoms represent a group with a more advanced disease that is more difficult to treat. These findings can assist clinicians in patient counselling, expectation management and decision-making about the timing of the intervention.Level of evidence: II.


[Collateral sprouting at end-to-side neurorrhaphy site demonstrated by nerve fibers micro-tease technique].
To develop a technique that can directly demonstrate collateral sprouting of intact nerve fibers at end-to-side neurorrhaphy site. Five Wistar adult rats were used in this study. The common peroneal nerves at one side were sectioned at the level of knee joint, and their distal ends were sutured to the tibial nerves after removal of a 1 mm-diameter window in the epineurium. Three months after the operation, the nerve segments at neurorrhaphy site and the normal tibial nerves at the contralateral site were harvested. The specimens were fixed in 10% formaldehyde and post-fixed in 1% osmium tetroxide, then macerated in glycerol. Single fiber was teased out in pure glycerol under an operative microscope, then transferred to a slide and observed under light microscope. The nerve segments at neurorrhaphy site and distal peroneal nerves were also harvested for histological evaluation. At the neurorrhaphy site, small nerve fibers sprouted from a donor nerve fiber near node's of Ranvier. While such phenomena were not found in normal tibial nerve. From the longitudinal section of neurorrhaphy site, bundles of nerve fibers ranged from tibial nerve to peroneal nerve were observed. Lots of regenerative nerve fibers emerged in distal peroneal nerve. The phenomena of collateral sprouting at end-to-side neurorrhaphy site can be demonstrated directly by nerve fiber micro-tease technique.


Repetitive extragenic palindromic PCR for study of Streptococcus mutans diversity and transmission in human populations.
Pulsed-field gel electrophoresis (PFGE) is considered the "gold standard" for molecular epidemiological study. Repetitive extragenic palindromic PCR (rep-PCR) is less time-consuming and more suitable for analyzing large numbers of bacterial strains in human populations. PFGE and rep-PCR provide comparable genotyping results for investigating Streptococcus mutans diversity and transmission.


Relaxation modulus of denture base resin reinforced with woven glass fibers.
This study examined the reinforcing effect of woven glass fibers on the relaxation modulus by three-point loading of denture base resin. Three sandwich-type flexure test specimens polymerized by heating were used, all 3 mm thick: glass fibers in compression (U type); glass fibers in the center (C type); and glass fibers in tension (L type). Unreinforced specimens (ACR) were produced as a control. The relaxation modulus after 8 sec from the start of stress (Er(8)) was calculated from stress relaxation curves. In the glassy state, the Er(8)s of the U and L types were greater than those of the ACR and C types. In the rubbery state (plateau region), the Er(8)s of the U and L types were greater than those of the ACR and C types. The woven glass fibers added to the U and L type specimens increased the relaxation modulus of PMMA near the oral temperature; moreover, they inhibited stress relaxation when the temperature increased.


[Persisting itching? Consider scabies].
Scabies is a common skin condition, which is accompanied by severe itching and a high degree of contagiousness. Rapid diagnosis and adequate treatment for the patient and his contacts is therefore important. Recognizing scabies is not always easy. Patients may present with atypical skin lesions. Persistent itching, more intense at night, and itching in relatives can then lead to the clinical (probable) diagnosis. Demonstration of the mite and/or eggs in the skin can confirm the diagnosis, either by dermatoscopy or direct microscopy of skin scrapings. Treatment consists of applying permethrin cream and/or taking ivermectin tablets. Although resistance to these agents has been suggested, treatment failure is mainly caused by failure to properly implement the treatment recommendations by the patient and/or his or her contacts. We present three patients with scabies in which the diagnosis and treatment could have been performed better.


Local olfactory interneurons provide the basis for neurochemical regionalization of olfactory glomeruli in crustaceans.
The primary olfactory centers of metazoans as diverse as arthropods and mammals consist of an array of fields of dense synaptic neuropil, the olfactory glomeruli. However, the neurochemical structure of crustacean olfactory glomeruli is largely understudied when compared to the insects. We analyzed the glomerular architecture in selected species of hermit crabs using immunohistochemistry against presynaptic proteins, the neuropeptides orcokinin, RFamide and allatostatin, and the biogenic amine serotonin. Our study reveals an unexpected level of structural complexity, unmatched by what is found in the insect olfactory glomeruli. Peptidergic and aminergic interneurons provide the structural basis for a regionalization of the crustacean glomeruli into longitudinal and concentric compartments. Our data suggest that local olfactory interneurons take a central computational role in modulating the information transfer from olfactory sensory neurons to projection neurons within the glomeruli. Furthermore, we found yet unknown neuronal elements mediating lateral inhibitory interactions across the glomerular array that may play a central role in modulating the transfer of sensory input to the output neurons through presynaptic inhibition. Our study is another step in understanding the function of crustacean olfactory glomeruli as highly complex units of local olfactory processing.


Imaging findings of intraventricular and ependymal lesions.
Intraventricular and ependymal lesions comprise a wide spectrum of tumoral, cystic, vascular, infectious and inflammatory disorders. With respect to tumoral and cystic diseases, the location, age and CT and MRI patterns are the main factors for diagnosis. The MRI findings of infectious diseases are supported by the clinical history, immune status and laboratory findings. Intracranial associated lesions may be very helpful for the diagnosis of Sturge-Weber, subependymal giant cell astrocytoma and systemic diseases, such as sarcoidosis and histiocytosis. Intraventricular vascular lesions are rare but present typical features on neuroimaging. The aim of this review is to provide a detailed description of these disorders with an emphasis on the key imaging findings and to generate a narrow differential diagnosis. We present a diagnostic approach based on the solid or cystic aspect of the intraventricular focal mass, its origin from the ventricular wall or choroid plexus and its location within the ventricular system. We also propose a differential diagnosis for ependymal dissemination: the ependymal enhancement may be due to ventriculitis from adjacent parenchymal lesions, the ependymal spread of tumors or infectious or inflammatory/systemic diseases.


Intravaginal ring delivery of the reverse transcriptase inhibitor TMC 120 as an HIV microbicide.
TMC 120 (Dapivirine) is a potent non-nucleoside reverse transcriptase inhibitor that is presently being developed as a vaginal HIV microbicide. To date, most vaginal microbicides under clinical investigation have been formulated as single-dose semi-solid gels, designed for application to the vagina before each act of intercourse. However, a clear rationale exists for providing long-term, controlled release of vaginal microbicides in order to afford continuous protection against heterosexually transmitted HIV infection and to improve user compliance. In this study we report on the incorporation of various pharmaceutical excipients into TMC 120 silicone, reservoir-type intravaginal rings (IVRs) in order to modify the controlled release characteristics of the microbicide. The results demonstrate that TMC 120 is released in zero-order fashion from the rings over a 28-day period and that release parameters could be modified by the inclusion of release-modifying excipients in the IVR. The hydrophobic liquid excipient isopropyl myristate had little effect on steady-state daily release rates, but did increase the magnitude and duration of burst release in proportion to excipient loading in the IVR. By comparison, the hydrophobic liquid poly(dimethylsiloxane) had little effect on TMC 120 release parameters. A hydrophilic excipient, lactose, had the surprising effect of decreasing TMC 120 burst release while increasing the apparent steady-state daily release in a concentration-dependent manner. Based on previous cell culture data and vaginal physiology, TMC120 is released from the various ring formulations in amounts potentially capable of maintaining a protective vaginal concentration. It is further predicted that the observed release rates may be maintained for at least a period of 1 year from a single ring device. TMC 120 release profiles and the mechanical properties of rings could be modified by the physicochemical nature of hydrophobic and hydrophilic excipients incorporated into the IVRs.


Terminal deoxynucleotidyl transferase (TdT) expression is associated with FLT3-ITD mutations in Acute Myeloid Leukemia.
The terminal deoxynucleotidyl transferase (TdT) is a DNA polymerase expressed in acute myeloid leukemias (AMLs), where it may be involved in the generation of NPM1 and FLT3-ITD mutations. We studied the correlations between TdT expression and FLT3-ITD or NPM1 mutations in primary AML samples, and the impact on patients' survival. TdT expression was analyzed in 143 adult AML patients by flow cytometry as percentage of positivity and mean fluorescence intensity (MFI) on blasts. TdT was positive in 49 samples (34.2%), with a median of 48% TdT-positivity (range 7-98) and a median MFI of 2.70 (range 1.23-30.54). FLT3-ITD and NPM1 mutations were present in 24 (16.7%) and 34 (23.7%) cases, respectively. Median TdT expression on blasts was significantly higher in FLT3-ITD+, as compared with FLT3-ITD- AMLs (median 8% vs 0% respectively, p = 0.035). NPM1 mutational status, FLT3-ITD allelic ratio, karyotype, and ELN risk groups, did not correlate with TdT expression or MFI on blasts. TdT + patients had poorer survival as compared to TdT-, but this result was not confirmed by the multivariable analysis, where ELN risk stratification as well as age and type of treatment remained independent prognostic factors for OS. In summary, our results support the possible implication of TdT enzyme in the generation of FLT3-ITD mutations in AML.


The effectiveness of AGV, Ex-PRESS, or trabeculectomy in the treatment of primary and secondary glaucoma: a systematic review and network meta-analysis.
In this study, we compared the efficacy of Ahmed, Ex-PRESS, and trabeculectomy to provide a reference for determining surgical schemes for glaucoma patients undergoing external drainage surgery in clinical practice. We performed a literature search for studies on the treatment of primary and secondary glaucoma with three types of external drainage surgery (Ahmed, Ex-PRESS, and trabeculectomy). As at April 24, 2021, seven electronic databases were searched for randomized controlled trials comparing any two of Ahmed, Ex-PRESS, and trabeculectomy in the treatment of glaucoma. The Cochrane tool was also adopted to evaluate the risk of bias in these trials. The relative risk (RR) with 95% confidence interval (CI), and weighted mean difference (WMD) were determined and compared indirectly using R software. A total of 14 randomized controlled trials were included in this study, involving 866 eyes of 808 patients. As for the intraocular pressure (IOP) after 3 months, trabeculectomy did not contribute to better improvement than Ahmed (WMD =0.014; 95% CI: -0.14-0.18) and Ex-PRESS (WMD =0.014; 95% CI: -0.072-0.097). However, there was a significant difference in the IOP 1 year between trabeculectomy and Ex-PRESS (WMD =0.097; 95% CI: 0.0080-0.18), with the latter achieving a favorable improvement effect. Meanwhile, the complete success (CS) of trabeculectomy was significantly lower than that of Ex-PRESS (RR =0.73; 95% CI: 0.57-0.93). In addition, Ex-PRESS was superior to Ahmed (WMD =-0.48; 95% CI: -0.89 to -0.084) in terms of a decreased number of post-operative medications. For glaucoma patients who are required to receive external drainage surgery, Ex-PRESS could achieve a significant effect on the IOP 1 year and CS, as well as a marked decrease in the number of post-operative medications used, compared with the other two types of surgery. In terms of the efficacy at least 1 year after surgery, Ex-PRESS should be one of the preferred methods for external drainage.


48-h Hypoxic exposure results in endothelium-dependent systemic vascular smooth muscle cell hyperpolarization.
Chronic hypoxia (CH) results in reduced sensitivity to vasoconstrictors in conscious rats that persists upon restoration of normoxia. We hypothesized that this effect is due to endothelium-dependent hyperpolarization of vascular smooth muscle (VSM) cells after CH. VSM cell resting membrane potential was determined for superior mesenteric artery strips isolated from CH rats (PB = 380 Torr for 48 h) and normoxic controls. VSM cells from CH rats studied under normoxia were hyperpolarized compared with controls. Resting vessel wall intracellular Ca(2+) concentration ([Ca(2+)](i)) and pressure-induced vasoconstriction were reduced in vessels isolated from CH rats compared with controls. Vasoconstriction and increases in vessel wall [Ca(2+)](i) in response to the alpha(1)-adrenergic agonist phenylephrine (PE) were also blunted in resistance arteries from CH rats. Removal of the endothelium normalized resting membrane potential, resting vessel wall [Ca(2+)](i), pressure-induced vasoconstrictor responses, and PE-induced constrictor and Ca(2+) responses between groups. Whereas VSM cell hyperpolarization persisted in the presence of nitric oxide synthase inhibition, heme oxygenase inhibition restored VSM cell resting membrane potential in vessels from CH rats to control levels. We conclude that endothelial derived CO accounts for persistent VSM cell hyperpolarization and vasoconstrictor hyporeactivity after CH.


Downregulation of FAK-related non-kinase mediates the migratory phenotype of human fibrotic lung fibroblasts.
Fibroblast migration plays an important role in the normal wound healing process; however, dysregulated cell migration may contribute to the progressive formation of fibrotic lesions in the diseased condition. To examine the role of focal-adhesion-kinase (FAK)-related non-kinase (FRNK) in regulation of fibrotic lung fibroblast migration, we examined cell migration, FRNK expression, and activation of focal adhesion kinase (FAK) and Rho GTPase (Rho and Rac) in primary lung fibroblasts derived from both idiopathic pulmonary fibrosis (IPF) patients and normal human controls. Fibrotic (IPF) lung fibroblasts have increased cell migration when compared to control human lung fibroblasts. FRNK expression is significantly reduced in IPF lung fibroblasts, while activation of FAK, Rho and Rac is increased in IPF lung fibroblasts. Endogenous FRNK expression is inversely correlated with FAK activation and cell migration rate in IPF lung fibroblasts. Forced exogenous FRNK expression abrogates the increased cell migration, and blocked the activation of FAK and Rho GTPase (Rho and Rac), in IPF lung fibroblasts. These data for the first time provide evidence that downregulation of endogenous FRNK plays a role in promoting cell migration through FAK and Rho GTPase in fibrotic IPF lung fibroblasts.


Modulation of human cardiac transient outward potassium current by EGFR tyrosine kinase and Src-family kinases.
The human cardiac transient outward K(+) current I(to) (encoded by Kv4.3 or KCND3) plays an important role in phase 1 rapid repolarization of cardiac action potentials in the heart. However, modulation of I(to) by intracellular signal transduction is not fully understood. The present study was therefore designed to determine whether/how human atrial I(to) and hKv4.3 channels stably expressed in HEK 293 cells are regulated by protein tyrosine kinases (PTKs). Whole-cell patch voltage-clamp, immunoprecipitation, western blotting, and site-directed mutagenesis approaches were employed in the present study. We found that human atrial I(to) was inhibited by the broad-spectrum PTK inhibitor genistein, the selective epidermal growth factor receptor (EGFR) kinase inhibitor AG556, and the Src-family kinases inhibitor PP2. The inhibitory effect was countered by the protein tyrosine phosphatase inhibitor orthovanadate. In HEK 293 cells stably expressing human KCND3, genistein, AG556, and PP2 significantly reduced the hKv4.3 current, and the reduction was antagonized by orthovanadate. Interestingly, orthovanadate also reversed the reduced tyrosine phosphorylation level of hKv4.3 channels by genistein, AG556, or PP2. Mutagenesis revealed that the hKv4.3 mutant Y136F lost the inhibitory response to AG556, while Y108F lost response to PP2. The double-mutant Y108F-Y136F hKv4.3 channels showed no response to either AG556 or PP2. Our results demonstrate that human atrial I(to) and cloned hKv4.3 channels are modulated by EGFR kinase via phosphorylation of the Y136 residue and by Src-family kinases via phosphorylation of the Y108 residue; tyrosine phosphorylation of the channel may be involved in regulating cardiac electrophysiology.


Teaching Diagnostic Reasoning to Medical Students: a four-step approach.
Diagnosis is fundamental to clinical medicine, and diagnostic errors are a serious public health problem. However, there is little consensus regarding the best approach to teaching diagnostic reasoning in medical schools. One approach ("pattern recognition") uses learned associations between patient symptoms and signs and human disorders to help experienced clinicians solve problems rapidly and efficiently. However, this approach may be ineffective when used by students with little clinical experience. Here we describe a four-step analytical approach to diagnosis that can be used by medical students before beginning clinical training. This approach complements the pattern recognition approach used by experts and can be used by students, residents in training, and attending physicians when confronting complex cases. The analytical approach also highlights critical basic science concept areas that support diagnostic reasoning and therefore warrant emphasis in medical school curricula. We propose introducing the analytical approach to medical students early in their training, coordinated with basic science instruction. Once students master relevant basic science concepts, they can use the analytical approach to diagnose disorders affecting one or more physiological systems, as a foundation for future clinical training.


Optical investigation of strong exciton localization in high Al composition AlxGa₁-xN alloys.
The exciton localization in wurtzite AlxGa₁-xN alloys with x varying from 0.41 to 0.63 has been studied by deep-ultraviolet photoluminescence (PL) spectroscopy and picosecond time-resolved PL spectroscopy. Obvious S-shape temperature dependence was observed indicating that the strong exciton localization can be formed in high Al composition AlxGa₁-xN alloys. It was also found that the Al composition dependence of exciton localization energy of AlGaN alloys is inconsistent with that of the excitonic linewidth. We contribute the inconsistency to the strong zero-dimensional exciton localization.


Milk ingestion stimulates net muscle protein synthesis following resistance exercise.
Previous studies have examined the response of muscle protein to resistance exercise and nutrient ingestion. Net muscle protein synthesis results from the combination of resistance exercise and amino acid intake. No study has examined the response of muscle protein to ingestion of protein in the context of a food. This study was designed to determine the response of net muscle protein balance following resistance exercise to ingestion of nutrients as components of milk. Three groups of volunteers ingested one of three milk drinks each: 237 g of fat-free milk (FM), 237 g of whole milk (WM), and 393 g of fat-free milk isocaloric with the WM (IM). Milk was ingested 1 h following a leg resistance exercise routine. Net muscle protein balance was determined by measuring amino acid balance across the leg. Arterial concentrations of representative amino acids increased in response to milk ingestion. Threonine balance and phenylalanine balance were both > 0 following milk ingestion. Net amino acid uptake for threonine was 2.8-fold greater (P < 0.05) for WM than for FM. Mean uptake of phenylalanine was 80 and 85% greater for WM and IM, respectively, than for FM, but not statistically different. Threonine uptake relative to ingested was significantly (P < 0.05) higher for WM (21 +/- 6%) than FM (11 +/- 5%), but not IM (12 +/- 3%). Mean phenylalanine uptake/ingested also was greatest for WM, but not significantly. Ingestion of milk following resistance exercise results in phenylalanine and threonine uptake, representative of net muscle protein synthesis. These results suggest that whole milk may have increased utilization of available amino acids for protein synthesis.


Central retinal artery occlusion after back surgery: a case report.
Central retinal artery occlusion can be caused by excessive extrinsic pressure on the eyeball during surgery. It is a well-known and severe complication after elective back surgery. The authors report a case of central retinal artery occlusion in an adult after posterior lumbar fusion in which a rectangular headrest was used.


Serum concentrations of Lp(a) and other lipoprotein parameters in heavy alcohol consumers.
The serum concentration of lipoprotein Lp(a) and of other lipoproteins of 64 individuals consuming more than 200 g of ethanol per day for several years was measured. 42 of the propositi with no clinical signs of alcoholic liver cirrhosis (ALC) had an apoAI/apoAII ratio twice as high as the control group. The apoAI values of the consumers of alcohol were higher and the HDL-C and Lp(a) levels were lower than those of the control group. 22 alcohol drinkers with ALC exhibited strikingly reduced values of all serum lipids and lipoproteins and the lowest Lp(a) concentrations of all. It is suggested that alcohol drinking lowers serum Lp(a) what might be an additional reason why this group of people seems to be a lower risk of atherosclerosis.


Ol-Prx 3, a member of an additional class of homeobox genes, is unimodally expressed in several domains of the developing and adult central nervous system of the medaka (Oryzias latipes).
Large-scale genetic screens for mutations affecting early neurogenesis of vertebrates have recently been performed with an aquarium fish, the zebrafish. Later stages of neural morphogenesis have attracted less attention in small fish species, partly because of the lack of molecular markers of developing structures that may facilitate the detection of discrete structural alterations. In this context, we report the characterization of Ol-Prx 3 (Oryzias latipes-Prx 3). This gene was isolated in the course of a large-scale screen for brain cDNAs containing a highly conserved DNA binding region, the homeobox helix-three. Sequence analysis revealed that this gene belongs to another class of homeobox genes, together with a previously isolated mouse ortholog, called OG-12 [Rovescalli, A. C., Asoh, S. & Nirenberg, M. (1996) Proc. Natl. Acad. Sci. USA 93, 10691-10696] and with the human SHOX gene [Rao, E., Weiss, B., Fukami, M., Rump, A., Niesler, B., et al. (1997) Nat. Genet. 16, 54-62], thought to be involved in the short-stature phenotype of Turner syndrome patients. These three genes exhibit a moderate level of identity in the homeobox with the other genes of the paired-related (PRX) gene family. Ol-Prx 3, as well as the PRX genes, are expressed in various cartilaginous structures of head and limbs. These genes might thus be involved in common regulatory pathways during the morphogenesis of these structures. Moreover, this paper reports a complex and monophasic pattern of Ol-Prx 3 expression in the central nervous system, which differs markedly from the patterns reported for the PRX genes, Prx 3 excluded: this gene begins to be expressed in a variety of central nervous system territories at late neurula stage. Strikingly, it remains turned on in some of the derivatives of each territory during the entire life of the fish. We hope this work will thus help identify common features for the PRX 3 family of homeobox genes.


Molecular Cloning, Recombinant Expression and Antifungal Activity of BnCPI, a Cystatin in Ramie (Boehmeria nivea L.).
Phytocystatins play multiple roles in plant growth, development and resistance to pests and other environmental stresses. A ramie (Boehmeria nivea L.) phytocystatin gene, designated as BnCPI, was isolated from a ramie cDNA library and its full-length cDNA was obtained by rapid amplification of cDNA ends (RACE). The full-length cDNA sequence (691 bp) consisted of a 303 bp open reading frame (ORF) encoding a protein of 100 amino acids with deduced molecular mass of 11.06 kDa and a theoretical isoelectric point (pI) of 6.0. The alignment of genome DNA (accession no. MF153097) and cDNA sequences of BnCPI showed that an intron (~104 bp) exists in the coding region. The BnCPI protein contains most of the highly conserved blocks including Gly⁵-Gly⁶ at the N-terminal, the reactive site motif QxVxG (Q49V50V51S52G53), the L79-W80 block and the [LVI]-[AGT]-[RKE]-[FY]-[AS]-[VI]-x-[EDQV]-[HYFQ]-N (L22G23R24 F25A26V27 D28D29H30 N31) block that is common among plant cystatins. BLAST analysis indicated that BnCPI is similar to cystatins from Glycine max (77%), Glycine soja (76%), Hevea brasiliensis (75%) and Ricinus communis (75%). The BnCPI was subcloned into expression vector pSmart-I and then overexpressed in Escherichia coli BL21 (DE3) as a His-tagged recombinant protein. The purified reBnCPI has a molecular mass of 11.4 kDa determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Purified reBnCPI can efficiently inhibit the protease activity of papain and ficin toward BANA (Nα-benzoyl-L-arginine-2-naphthyamide), as well as the mycelium growth of some important plant pathogenic fungi. The data further contribute to our understanding of the molecular functions of BnCPI.


Systematic study of the substituted active C-terminus of hirudin.
Hirudin, a potent clinical thrombin inhibitor from Hirudo medicinalis, consists of 65 amino acids in a single chain. In this paper, we systematically synthesize a series of C-terminal (desulfo hirudin45-65) peptides substituted by 20 natural L-amino acids via the Multipin method. The resulting peptide library is subsequently screened using an alpha-thrombin-mediated fibrinogen clotting assay and alpha-thrombin-induced amidolytic hydrolysis assay.


Synthesis and characterization of boranate ionic liquids (BILs).
Straightforward access to hydridoborate-based ionic liquids (BILs) is provided. They fall into a barely developed area of research and are of interest as, for example, reagents for organic synthesis. A series of pure [BH(4)](-) ILs with 1-butyl-2,3-dimethylimidazolium (BMMIM), 1-ethyl-3-methylimidazolium (EMMIM), 1-propyl-1-methylpiperidinium (PropMPip), and1-butyl-1-methylpyrrolidinium (BMP) cations were prepared. All synthesized ILs are well soluble in CH(2)Cl(2). We developed a procedure that gives clean products with correct elemental analyses. In contrast to earlier reports, which when conducted by us yielded only mixtures of the boranate anion with major halide contamination (maximum [BH(4)](-) content: 77.5 %). These materials can be viewed as the starting material for the (hypothetical) hydrogen-storage redox shuttling sequence between [BH(4)](-) and [B(12)H(12)](2-), in which the triboranate anion [B(3)H(8)](-) is a formal intermediate. Here we also developed a facile route to [B(3)H(8)](-) ILs with [BMMIM](+), [EMMIM](+), [PropMPip](+), and [NBu(4)](+), in which Na[BH(4)] reacts in situ (enhanced by ultrasound) with the solvent CH(2)Cl(2) as the oxidizing agent to give the triboranate IL in high yield and purity according to the equation: 3 [BH(4)](-)+2 CH(2)Cl(2)+[Cat](+)→[B(3)H(8)](-)[Cat](+)+H(2)+2 CH(3)Cl+2 Cl(-). We further investigated this reaction path by additional NMR spectroscopic experiments, powder-XRD analysis, and quantum chemical DFT calculations.


Steroid receptor coactivator expression throughout the menstrual cycle in normal and abnormal endometrium.
The endometrium of reproductive aged women undergoes cyclic developmental changes in preparation for implantation in response to estrogen and progesterone. These steroids and their receptors are tightly regulated throughout the menstrual cycle, and their actions are facilitated by the presence of steroid receptor coactivators of the p160 family. In this study using immunohistochemistry and Western blot analysis, we characterize the expression patterns of three coactivators, steroid receptor coactivator-1, amplified in breast cancer-1 (AIB1), and transcriptional intermediary factor-2 in human endometrium obtained prospectively from normal fertile women throughout the menstrual cycle. With the exception of glandular AIB1, which increased in the late secretory phase, none of the coactivators changed significantly during the menstrual cycle. We compared coactivator expression patterns in fertile endometrium to the endometrium of anovulatory (proliferative; n = 3) and clomiphene-induced ovulatory (secretory; n = 13) women with polycystic ovarian syndrome (PCOS), a group that have a higher likelihood of developing estrogen-induced endometrial hyperplasia and cancer. To control for the effect of clomiphene citrate, an additional group was included consisting of ovulatory women treated with clomiphene citrate for "male factor" infertility. Compared with both fertile and infertile controls, PCOS women exhibited elevated levels of AIB1 and transcriptional intermediary factor-2 expression in both epithelial and stromal cells. We postulate that increased coactivator expression may render the endometrium more sensitive to estrogen. In support of this, we describe an increased expression of ERalpha (an estrogen-induced gene product) during the menstrual cycle in PCOS endometrium compared with fertile controls. In summary, we demonstrate that the expression of p160 coactivators are regulated in endometrium during the menstrual cycle in normal fertile women but are overexpressed in the endometrium of women with PCOS. Based on these findings, we suggest a possible mechanism to explain the poor reproductive performance observed in PCOS and the increased incidence of endometrial hyperplasia and cancer noted in this group of women.