RAS signaling in colorectal carcinomas through alteration of RAS, RAF, NF1, and/or RASSF1A.
More than half of all colorectal carcinomas are known to exhibit an activated mitogen-activated protein kinase pathway. The NF1 gene, a negative regulator of KRAS, has not previously been examined in a series of colorectal cancer. In the present study, primary colorectal carcinomas stratified according to microsatellite instability status were analyzed. The whole coding region of NF1 was analyzed for mutations using denaturing high-performance liquid chromatography and sequencing, and the copy number alterations of NF1 were examined using multiple ligation-dependent probe amplification and real-time polymerase chain reaction. The mutational hot spots in KRAS and BRAF were sequenced, and promoter hypermethylation status of RASSF1A was assessed with a methylation-specific polymerase chain reaction. One sample had two missense mutations in NF1, whereas nine additional tumors had intronic mutations likely to affect exon splicing. Interestingly, 8 of these 10 tumors were microsatellite-unstable. Four other tumors showed a duplication of NF1. Mutations in KRAS and BRAF were mutually exclusive and were present at 40% and 22%, respectively. RASSF1A was hypermethylated in 31% of the samples. We show that the RAS signaling network is extensively dysregulated in colorectal carcinomas, because more than 70% of the tumors had an alteration in one or more of the four examined components.


Endogenous methicillin resistant staphylococcus aureus as a source of post-operative surgical site infections.
To determine frequency of Endogenous Methicillin Resistant Staphylococcus aureus (MRSA) in pre-operative patients and its frequency in Surgical Site Infections (SSIs) post operatively. It was a descriptive cross sectional conducted at Department of Microbiology, Fauji Foundation Hospital Rawalpindi (FFH), Pakistan. Samples were collected from 1st November-31st May 2018. Total 75 samples were collected during the period. Consecutive non-probability sampling technique was utilized. Specimens were collected from nose, axilla and groin of preoperative patients. Methicillin Resistant Staphylococcus aureus was identified if only isolated from these sites. Patients were followed till his/her discharge from the hospital and if they developed infection post operatively, pus specimen from infected site was also collected and identified. Out of 75 specimen preoperatively, 11(14.7%) were identified as endogenous MRSA. From these, 33(44%) developed Surgical site infections (SSIs), among them 19(57%) were MRSA (09 endogenous, 10 exogenous), 7(21.2%) were Escherichia coli, 3(9.1%) were Klebsiella pneumoniae, 3(9.1%) were Enterococcus faecalis and 1(3%) was Methicillin Sensitive Staphylococcus aureus. The results of this study determined that Endogenous Methicillin Resistant Staphylococcus aureus (MRSA) could be isolated from patients going for surgery if microbiological screening was done at the time of admission. This could prevent patients from Surgical Site Infection Post operatively by these endogenous MRSA. This search and wipe out strategy is able to curtail the events of outbreak, reduce hospital stay and decrease budget of the hospital by providing guidance in choice of empirical therapy for infection.


Comparative efficacy of monophasic and biphasic waveforms for transthoracic cardioversion of atrial fibrillation and atrial flutter.
Transthoracic cardioversion fails to restore sinus rhythm in 6% to 33% of patients with atrial fibrillation. This study sought to determine the relative efficacy of biphasic waveforms compared with monophasic waveforms in the treatment of atrial arrhythmias. A total of 912 patients underwent 1022 transthoracic cardioversions between May 2000 and December 2001. A monophasic damped sine waveform was used in the first 304 cases, and a rectilinear biphasic defibrillator was used in the next 718 cases. Use of a biphasic waveform was associated with 94% success in conversion to sinus rhythm compared with 84% with a monophasic waveform (P < .001). The cumulative energy required to restore sinus rhythm was lower with biphasic shocks in both atrial fibrillation and atrial flutter groups (554 +/- 413 J for monophasic vs 199 +/- 216 J for biphasic shocks in the atrial fibrillation group, P < .001; 251 +/- 302 J vs 108 +/- 184 J, respectively, in the atrial flutter group, P < .001). In a multivariate analysis, use of a biphasic shock was associated with a 3.9-fold increase in success of cardioversion. When used to cardiovert atrial arrhythmias, the rectilinear biphasic waveform was associated with higher success rates and lower cumulative energies than the monophasic damped sine waveform.


Bronchoalveolar lavage in idiopathic interstitial lung diseases.
Bronchoalveolar lavage (BAL) is useful for diagnosing various interstitial lung diseases (ILDs) and monitoring ILD during treatment. The ability to detect specific agents or substances by using BAL is especially helpful in determining whether idiopathic ILD has a background causality. BAL in combination with other intensive examinations may enable the definitive diagnosis of an idiopathic ILD. Among the idiopathic ILDs of concern, this article focuses on idiopathic interstitial pneumonia (IIP) and cryptogenic organizing pneumonia (COP). IIP and COP are classified together as idiopathic interstitial pneumonia (IIP), an integrated clinicoradiographic pathological disease entity. BAL has identified two points important for differentiating the disease entity: a paucity of lymphocytes appears in the BAL fluid of patients with idiopathic pulmonary fibrosis (IPF), and lymphocytosis with a decreased CD4:CD8 ratio appears in patients with COP. These findings can be useful, in combination with high-resolution computed tomographic (HRCT) data, for selecting a favorable treatment option. On the other hand, these rules cannot be applied to IP associated with collagen vascular disease (CVD). Furthermore, some IIP patients may manifest features of CVD during the clinical course after the detection of IP (interstitial pneumonia). Thus the definite role of BAL cell profiles remains to be determined.


Green Pharmaceutical Analysis of Drugs Coformulated with Highly Different Concentrations Using Spiking and Manipulation of Their Ratio Spectra.
Introducing green analysis to pharmaceutical products is considered a significant approach to preserving the environment. This method can be an environmentally friendly alternative to the existing methods, accompanied by a validated automated procedure for the analysis of a drug with the lowest possible number of samples. Different simple spectrophotometric methods were developed for the simultaneous determination of empagliflozin (EG) and metformin (MT) by manipulating their ratio spectra in their application on a recently approved pharmaceutical combination, Synjardy tablets. A spiking technique was used to increase the concentration of EG in samples prepared from the tablets to allow for the simultaneous determination of EG with MT without prior separation. Validation parameters according to International Conference on Harmonization guidelines were acceptable over a concentration range of 2-12 μg/mL for both drugs using derivative ratio and ratio subtraction coupled with extended ratio subtraction. The optimized methods were compared using one-way analysis of variance and proved to be suitable as ecofriendly approaches for industrial QC laboratories.


[Importance of the components of the complex ester group for the action of acetylcholine and dicarboxylic acid dicholine esters].
The substitution of ether oxygen by a methylene group in the acetylcholine molecule sharply decreases the muscarinic action, while the substitution of the carbonyl group by a methylene one reduces the nicotinic properties. Sebacoyldicholine possesses only nicotinic properties. The replacement of its ether oxygens by methylene groups lowers the stimulant action on the frog rectus abdominis muscle by two orders of magnitude, whereas the stimulant action on the cat sympathetic ganglion is 30-fold as decreased. The blocking action on the isolated rat diaphragm becomes 30-fold less potent, but the blocking action on the cat skeletal muscle remains as strong as that of sebacoyldicholine after cholinesterase inhibition. The replacement of sebacoyldicholine carbonyl groups makes the compound elicit a noncompetitive cholinolytic action on the frog muscle, while the blocking action on the cat skeletal muscle becomes as weak as that exerted by hexadecamethylene-bis-trimethyammonium.


[Resection and reconstruction of the retrohepatic vena cava in combination with liver resections].
Liver resection combined with the resection and reconstruction of the vena cava represents the only potential curative therapy for malignant hepatic tumors with invasion of the vena cava. We performed a liver resection with segmental replacement of the retrohepatic vena cava by synthetic grafts in 29 patients. In three cases, the additional presence of central involvement of all three hepatic veins required ex situ tumor resection. Four patients underwent a simultaneous exstirpation of the primary tumor (kidney or suprarenals). The remaining hepatic veins were reimplanted into the graft in three cases, and in two cases the renal veins were reimplanted. There was no perioperative mortality. A distal arteriovenous fistula was not applied. Five patients revealed postoperative transient liver insufficiency, requiring temporary dialysis in three cases. Two of these patients developed a transient multiorgan failure with the need of mechanical ventilation. 18 patients died during the course of follow-up, 17 of these cases due to recurrent metastases of the primary disease. Infection or thrombosis of the prosthetic vascular graft have not been observed. Beside tumor exstirpation, extended liver resection and concomitant vena cava replacement may prevent embolism as well as the obstruction of the vena cava with lower extremity swelling and the possibility of developing a Budd Chiari syndrome. We were able to achieve a long-term survival for surgically treated patients even in cases with advanced tumor stages.


Pancreas transplantation with portal venous drainage with an emphasis on technical aspects.
Advances in surgical techniques and clinical immunosuppression have led to improving results in vascularized pancreas transplantation. Most pancreas transplants are performed with enteric exocrine drainage and systemic venous delivery of insulin (systemic-enteric technique) although bladder drainage (systemic-bladder technique) remains a viable option. To improve the physiology of pancreas transplantation, an innovative technique of portal venous delivery of insulin and enteric drainage of the exocrine secretions (portal-enteric technique) was developed and refined over the past 27 yr. However, the potential of portal-enteric pancreas transplantation has never been fully realized as it is currently performed in only 18% of simultaneous pancreas-kidney/sequential pancreas after kidney and 10% of pancreas-alone transplants with enteric drainage. A number of studies have demonstrated no major or consistent differences in outcomes for bladder-drained or enteric-drained pancreas transplants with either portal or systemic venous drainage although some studies suggest purported metabolic and immunologic advantages associated with portal venous delivery of insulin. The purpose of this study is to review the existing literature on portal-enteric pancreas transplantation with an emphasis on surgical aspects and technical modifications/nuances that have been introduced with time and experience.


Effect of supplements with lactic acid bacteria and oligofructose on the intestinal microflora during administration of cefpodoxime proxetil.
Thirty healthy volunteers in three groups participated in a study of the effect on the intestinal microflora of oral supplementation with Bifidobacterium longum, Lactobacillus acidophilus and oligofructose, an indigestible oligosaccharide, during oral administration of cefpodoxime proxetil bd for 7 days. Those in group A also received an oral supplement with c.1011 cfu of B. longum BB 536 and L. acidophilus NCFB 1748 and 15 g oligofructose daily, those in group B received a supplement with oligofructose only and those in group C received placebo, for 21 days. In all three groups there was a marked decrease in aerobic microorganisms, involving mainly a rapid and almost complete disappearance of Escherichia coli (P: < 0.05) during antimicrobial administration and, thereafter, an overgrowth of enterococci (P: < 0.05). The number of intestinal yeasts also increased significantly (P: < 0.05) in groups A and B over the same period. There was a dramatic decrease in anaerobic microorganisms on day 4 of administration, mainly caused by loss of bifidobacteria (P: < 0.05) in all groups. The number of lactobacilli also decreased but was significantly higher in group A than in group C at the end of cefpodoxime proxetil administration. Clostridium difficile was found in only one person from group A, but six persons each in groups B and C. Of the bifidobacterial strains isolated from the faecal samples in group A, one was similar to the strain of B. longum administered, but most volunteers were colonized by several different strains of B. longum during the investigation period. The administered strain of L. acidophilus was recovered from six patients in group A.


Peliosis hepatis in recipients of renal transplants.
Peliosis hepatis, an uncommon liver lesion characterised by blood-filled cavities bordered by hepatocytic plates, was found in 12 patients three to 17 months after renal transplantation. Hepatomegaly and portal hypertension were present in five of the six patients with major peliosis hepatis, and were absent in the other six with minor hepatic lesions. Alterations of centrilobular vein walls in some of these patients suggest that peliosis hepatis could be the result of a blockade or liver blood outflow at the junctions of sinusoids and centrilobular veins. The cause of these alterations might be azathioprine.


[Hospital Mortality in Parkinson's Disease: Retrospective Analysis in a Portuguese Tertiary Centre].
Parkinson's disease is associated with high hospital mortality. Male gender, late age at onset, higher disability and the coexistence of cognitive impairment or depression have been suggested to be risk factors of death. Pneumonia and cardiovascular diseases are the leading causes of death. To characterize the mortality (causes of hospital admission and death) of Parkinson's disease patients in a tertiary hospital, as well as demographic and clinical characteristics. Identification of hospital admissions of Parkinson's disease patients that resulted in inpatient death between 2008 and 2014. Retrospective review of medical files and inclusion of patients with disease clinically confirmed by a neurologist. Assessment of causes of death and demographic and clinical characteristics of patients. 1525 hospital admissions of Parkinson's disease patients were identified, of which 150 resulted in death. Of these, 52 patients met the inclusion criteria. Mean age at onset of Parkinson's disease symptoms was 66.8 years (± 8.7) and mean duration of disease was 12.5 years (± 7.9). Sixty-five percent of patients were in stages 4-5 of the Hoehn and Yahr scale. Thirty-three patients (63%) had dementia and eleven (21%) had depression. Infections were the leading cause of death (respiratory in 63% of cases). Similarly to literature, pneumonia was the leading cause of hospital death and most patients presented advanced disease stage and dementia. Contrarily to other studies, life expectancy was not reduced and cardiovascular diseases and trauma were not causes of death in our population. This is the first Portuguese mortality study in Parkinson's disease. Pneumonia is the leading cause of death. Advanced disease stage and dementia were common features in these patients.


Surgical Reduction and Fixation of Tibial Spine Fractures in Children: Arthroscopic Suture Fixation.
The pediatric tibial spine fracture can be considered the equivalent of the pediatric anterior cruciate ligament (ACL) rupture, especially considering that failure to manage these injuries appropriately could result in substantial knee pathology and long-term functional failure. Arthroscopy-assisted internal fixation of the tibial spine fracture minimizes the patient's hospital stay compared with that associated with an open arthrotomy approach, but the postoperative rehabilitation may be paramount to a successful outcome. Compared with conservative management, surgical reduction and fixation of these fractures decreases the risks of comorbidities related to meniscal tissue entrapment, the risk of anterior impingement, and the need for revision treatment. Repair should be considered for fractures displaced >5 mm at their maximum, given the risk of revision when these cases are treated without surgery. After reduction and tentative fixation, the fracture can be managed with either suture or screw fixation. For suture fixation, the surgical steps include (1) creation of peripatellar arthroscopy portals; (2) evacuation of the hematoma and clearing of the fracture bed of meniscus and callus; (3) reduction, with particular attention paid to articular surface extension if present, and tentative fixation with an ACL tibial guide and placement of threaded Kirschner wire; (4) drilling of bone tunnels for suture passage using the tibial guide juxtaposed to the ACL insertion (both medially and laterally, in turn); (5) passing a suture of choice (nonabsorbable type requiring intraoperative fluoroscopy to evaluate tunnel placement during the previous step); (6) passage of suture both posterior and anterior to the ACL fibers; (7) removal of the tentative threaded Kirschner wire; (8) tying of the posterior suture over a bone bridge; (9) tying of the anterior suture over the same bone bridge; and (10) standard arthroscopy portal closure and placement of dressings including immobilization of the knee. For screw fixation, the surgical steps are slightly different, with Step 3 followed by (4) placement of the guidewire (if a cannulated screw is being used), (5) intraoperative assessment of the fracture reduction and drilling of the screw track perpendicular to the vector of the ACL pull and tentative threaded Kirschner wire fixation, (6) measurement and placement of the final screw implant, (7) removal of the tentative Kirschner wire fixation prior to final setting of the screw, and (8) standard arthroscopy portal closure and placement of dressings including immobilization of the knee. Radiographic and functional outcomes are excellent with either fixation technique in this pediatric population.


Mathematical model of 137Cs migration in soil: analysis of observations following the Chernobyl accident.
The applicability of traditional mathematical models is studied on the basis of 3 y of observation of the vertical migration of 137Cs fallout after the Chernobyl accident. The most accurate description of the dependence of the radionuclide concentration on the depth of the soil layer is given by a lognormal distribution. The parameters of this distribution are determined and shown to be a solution to the Fokker-Planck equation, a special case of which is the diffusion-convection transport equation.


Erk and PI-3 kinase are necessary for collagen binding and actin reorganization in corneal epithelia.
It was recently shown that phosphatidylinositol-(PI)3 kinase is upregulated in wounded rabbit corneal epithelia. Extracellular signal-regulated kinase (erk)-1 and -2 proteins and PI-3 kinase were activated in embryonic corneal epithelia after 1-hour stimulation by type I collagen. In the current investigation specific inhibitors of PI-3 kinase and mitogen-activated kinase-kinase (MEK-1 kinase) were used to determine the role of these signaling molecules in actin reorganization and collagen binding to isolated sheets of corneal epithelial tissue. Effects of specific PI-3 kinase and MEK-1 inhibitors (LY294002, PD98059, respectively) were investigated in embryonic corneal epithelial tissues. Avian embryonic corneal epithelia were isolated as tissue sheets, organ cultured in the presence of these specific inhibitors, and stimulated with type I collagen. The tissues were evaluated for collagen-stimulated actin reorganization, erk-1 and -2 and PI-3 kinase activity, total filamentous actin accumulation, and collagen binding. The MEK-1 inhibitor PD98059 decreased erk-1 and -2 phosphorylation and blocked actin reorganization in a dose-dependent manner. The PI-3 kinase 85-kDa subunit was decreased 25% in LY294002-treated tissue, and collagen binding also decreased significantly in tissues treated with MEK-1 and PI-3 kinase inhibitors compared with control tissues. In addition, both inhibitors blocked actin cortical mat reorganization. CONCLUSIONS; PI-3 kinase and erk-1 and -2 signaling pathways are activated and necessary for collagen binding and integrin-mediated actin reorganization in embryonic avian corneal epithelium.


Daily rhythms of catalase and glutathione peroxidase expression and activity are endogenously driven in the hippocampus and are modified by a vitamin A-free diet.
Alterations in enzymatic antioxidant defense systems lead to a deficit of cognitive functions and altered hippocampal synaptic plasticity. The objectives of this study were to investigate endogenous rhythms of catalase (CAT) and glutathione peroxidase (GPx) expression and activity, as well as CREB1 mRNA, in the rat hippocampus, and to evaluate to which extent the vitamin A deficiency could affect those temporal patterns. Rats from control and vitamin A-deficient (VAD) groups received a diet containing 4000 IU of vitamin A/kg diet, or the same diet devoid of vitamin A, respectively, during 3 months. Rats were maintained under 12-hour-dark conditions, during 10 days before the sacrifice. Circadian rhythms of CAT, GPx, RXRγ, and CREB1 mRNA levels were determined by reverse transcriptrase polymerase chain reaction in hippocampus samples isolated every 4 hours during a 24-hour period. CAT and GPx enzymatic activities were also determined by kinetic assays. Regulatory regions of clock and antioxidant enzymes genes were scanned for E-box, RXRE, and CRE sites. E-box, RXRE, and CRE sites were found on regulatory regions of GPx and CAT genes, which display a circadian expression in the rat hippocampus. VAD phase shifted CAT, GPx, and RXRγ endogenous rhythms without affecting circadian expression of CREB1. CAT and GPx expression and enzymatic activity are circadian in the rat hippocampus. The VAD affected the temporal patterns antioxidant genes expression, probably by altering circadian rhythms of its RXR receptors and clock factors; thus, it would impair the temporal orchestration of hippocampal daily cognitive performance.


A qualitative exploration of young adult smokers' responses to novel tobacco warnings.
Despite reduced smoking among adolescents, smoking prevalence peaks among young adults aged 18-30, many of whom believe themselves exempt from the health risks of smoking shown in warning labels. We explored how young adult smokers perceived warnings featuring proximal risks, and whether these encouraged cessation more effectively than traditional health messages. We conducted in-depth interviews with 17 young adult smokers and explored their perceptions of current warnings as well as novel warnings representing short-term health consequences; immediate social risks, and tobacco's toxicity (denormalizing tobacco as an everyday product). We used a thematic analysis approach to explore how participants rationalized existing warnings and interpreted the novel messages. Participants considered the immediate social and physiological benefits they gained from smoking outweighed the distal risks shown in health warnings, which they regarded as improbable and irrelevant. Of the novel warnings, those presenting immediate social risks altered the balance of gains and losses young adults associated with smoking; however, those presenting short-term health risks or depicting tobacco as a toxin were less effective. Participants regarded warnings featuring proximal social risks as more salient and they were less likely to rationalise these as irrelevant. Social risk messages merit further investigation to examine their potential as a complement to traditional health warnings.


Comparison of two major forms of the Shigella virulence plasmid pINV: positive selection is a major force driving the divergence.
All Shigella and enteroinvasive Escherichia coli (EIEC) strains carry a 230-kb virulence plasmid (pINV) which is essential for their invasiveness. There are two sequence forms, pINV A and pINV B, of the plasmid (R. Lan, B. Lumb, D. Ryan, and P. R. Reeves, Infect. Immun. 69:6303-6309, 2001), and the recently sequenced pINV plasmid from Shigella flexneri serotype 5 is a pINV B form. In this study we sequenced the majority of the coding region of the pINV A form from S. flexneri serotype 6 other than insertion sequence or related sequences and compared it with the pINV B form. More than half of the genes sequenced appear to be under positive selection based on their low ratio of synonymous to nonsynonymous substitutions. This high proportion of selected differences indicates that the two pINV forms have functional differences, and comparative studies of pathogenicity in different Shigella-EIEC strains could be informative. There are also genes absent in the S. flexneri serotype 6 plasmid, including the sepA gene encoding serine protease, the major secreted protein of S. flexneri serotype 2a, and the stbAB genes, which encode one of the two partition systems found in S. flexneri serotype 5. The incompatibility of the two pINV forms appears to be due to either small differences in the mvpAT postsegregational killing system or the presence of an unknown system in pINVA.


Nanoscale patterning of a self-assembled monolayer by modification of the molecule-substrate bond.
The intercalation of Cu at the interface of a self-assembled monolayer (SAM) and a Au(111)/mica substrate by underpotential deposition (UPD) is studied as a means of high resolution patterning. A SAM of 2-(4'-methylbiphenyl-4-yl)ethanethiol (BP2) prepared in a structural phase that renders the Au substrate completely passive against Cu-UPD, is patterned by modification with the tip of a scanning tunneling microscope. The tip-induced defects act as nucleation sites for Cu-UPD. The lateral diffusion of the metal at the SAM-substrate interface and, thus, the pattern dimensions are controlled by the deposition time. Patterning down to the sub-20 nm range is demonstrated. The difference in strength between the S-Au and S-Cu bond is harnessed to develop the latent Cu-UPD image into a patterned binary SAM. Demonstrated by the exchange of BP2 by adamantanethiol (AdSH) this is accomplished by a sequence of reductive desorption of BP2 in Cu free areas followed by adsorption of AdSH. The appearance of Au adatom islands upon the thiol exchange suggests that the interfacial structures of BP2 and AdSH SAMs are different.


Arthropod 5-HT2 receptors: a neurohormonal receptor in decapod crustaceans that displays agonist independent activity resulting from an evolutionary alteration to the DRY motif.
The stomatogastric nervous system (STNS) is a premiere model for studying modulation of motor pattern generation. Whereas the cellular and network responses to monoamines have been particularly well characterized electrophysiologically, the transduction mechanisms that link the different monoaminergic signals to specific intracellular responses are presently unknown in this system. To begin to elucidate monoaminergic signal transduction in pyloric neurons, we used a bioinformatics approach to predict the existence of 18 monoamine receptors in arthropods, 9 of which have been previously cloned in Drosophila and other insects. We then went on to use the two existing insect databases to clone and characterize the 10th putative arthropod receptor from the spiny lobster, Panulirus interruptus. This receptor is most homologous to the 5-HT2 subtype and shows a dose-dependent response to 5-HT but not to any of the other monoamines present in the STNS. Through a series of pharmacological experiments, we demonstrate that this newly described receptor, 5-HT2betaPan, couples with the traditional G(q) pathway when expressed in HEK293 cells, but not to G(s) or G(i/o). Moreover, it is constitutively active, because the highly conserved DRY motif in transmembrane region 3 has evolved into DRF. Site-directed mutagenesis that reverts the motif back to DRY abolishes this agonist-independent activity. We further demonstrate that this receptor most likely participates in the modulation of stomatogastric motor output, because it is found in neurites in the synaptic neuropil of the stomatogastric ganglion as well as in the axon terminals at identified pyloric neuromuscular junctions.


Extensive next-generation sequencing analysis in chronic lymphocytic leukemia at diagnosis: clinical and biological correlations.
In chronic lymphocytic leukemia (CLL), next-generation sequencing (NGS) analysis represents a sensitive, reproducible, and resource-efficient technique for routine screening of gene mutations. We performed an extensive biologic characterization of newly diagnosed CLL, including NGS analysis of 20 genes frequently mutated in CLL and karyotype analysis to assess whether NGS and karyotype results could be of clinical relevance in the refinement of prognosis and assessment of risk of progression. The genomic DNA from peripheral blood samples of 200 consecutive CLL patients was analyzed using Ion Torrent Personal Genome Machine, a NGS platform that uses semiconductor sequencing technology. Karyotype analysis was performed using efficient mitogens. Mutations were detected in 42.0 % of cases with 42.8 % of mutated patients presenting 2 or more mutations. The presence of mutations by NGS was associated with unmutated IGHV gene (p = 0.009), CD38 positivity (p = 0.010), risk stratification by fluorescence in situ hybridization (FISH) (p < 0.001), and the complex karyotype (p = 0.003). A high risk as assessed by FISH analysis was associated with mutations affecting TP53 (p = 0.012), BIRC3 (p = 0.003), and FBXW7 (p = 0.003) while the complex karyotype was significantly associated with TP53, ATM, and MYD88 mutations (p = 0.003, 0.018, and 0.001, respectively). By multivariate analysis, the multi-hit profile (≥2 mutations by NGS) was independently associated with a shorter time to first treatment (p = 0.004) along with TP53 disruption (p = 0.040), IGHV unmutated status (p < 0.001), and advanced stage (p < 0.001). Advanced stage (p = 0.010), TP53 disruption (p < 0.001), IGHV unmutated status (p = 0.020), and the complex karyotype (p = 0.007) were independently associated with a shorter overall survival. At diagnosis, an extensive biologic characterization including NGS and karyotype analyses using novel mitogens may offer new perspectives for a better refinement of risk stratification that could be of help in the clinical management of CLL patients.


Women's knowledge and beliefs towards vaccination for influenza during pregnancy in Turkey and underlying factors of misinformation: a single-centre cross-sectional study.
The aim of the study was to evaluate the knowledge and perceptions of the pregnant women presenting to our hospital for seasonal vaccination for influenza and to determine the factors associated with it. In this cross-sectional study pregnant woman presenting to our hospital between October 2018 and March 2019 were evaluated. A non-validated, well-detailed questionnaire addressing the vaccination rates, participants' perceptions about the facts behind the vaccination for influenza and the factors associated with refusal of vaccination was performed. Women's knowledge level provided by their healthcare providers was also questioned. A total of 250 participants were included in the study. The average age of the patients was 28.85 ± 5.42 years (range 18-43); and the average pregnancy week was 19 ± 9.75. It was determined that 98% (n = 245) of the participants did not have any vaccinations before, and 98.8% (n = 247) did not have any vaccination during their current pregnancy. 65.2% (n = 163) of the participants did not know that the vaccination for influenza was safe in pregnancy; and 64% (n = 160) did not know that the vaccination for influenza was recommended in pregnancy. The most frequent responses given by the participants to justify their refusal for the vaccination was "my doctor was against" and "it can be harmful to my baby" (25.6% and 24%, respectively). It was determined that 98.4% (n = 246) of the participants were not recommended about the vaccination for influenza by any healthcare centres; and 92.8% (n = 232) did not receive any information on vaccination for influenza. The knowledge of the participants on vaccination for influenza was inadequate and had misconceptions. The inadequacy of healthcare employees, government institutions and the media may have played roles in this outcome. The reasons underlying the inadequacy of the healthcare providers on vaccination for influenza may be questioned.


Serum concentrations and expressions of serum amyloid A and leptin in adipose tissue are interrelated: the Genobin Study.
Serum amyloid A (SAA) is a novel link between increased adipose tissue mass and low-grade inflammation in obesity. Little is known about the factors regulating its serum concentration and mRNA levels. We investigated the association between SAA and leptin in obese and normal weight subjects and analyzed the effect of weight reduction on serum SAA concentration and gene expression in adipose tissue of the obese subjects. Seventy-five obese subjects (60+/-7 years, body mass index (BMI) 32.9+/-2.8 kg/m(2), mean+/-s.d.) with impaired fasting plasma glucose or impaired glucose tolerance and other features of metabolic syndrome, and 11 normal weight control subjects (48+/-9 years, BMI 23.7+/-1.9 kg/m(2)) were studied at the baseline. Twenty-eight obese subjects underwent a 12-week intensive weight reduction program followed by 5 months of weight maintenance. Blood samples and abdominal s.c. adipose tissue biopsies were taken at the baseline and after the follow-up. Gene expression was studied using real-time quantitative PCR. The gene expressions in women and serum concentrations of leptin and SAA were interrelated independently of body fat mass in the obese subjects (r=0.54, P=0.001; r=0.24, P=0.039 respectively). In multiple linear regression analyses, leptin mRNA explained 38% of the variance in SAA mRNA (P=0.002) in the obese women. Weight loss of at least 5% increased SAA mRNA expression by 48 and 36% in men and women, but serum SAA concentrations did not change. The association between SAA and leptin suggests an interaction between these two adipokines, which may have implications in inflammatory processes related to obesity and the metabolic syndrome.


A national UK audit of suprapubic catheter insertion practice and rate of bowel injury with comparison to a systematic review and meta-analysis of available research.
Limited data exist on the risks of complications associated with a suprapubic catheter (SPC) insertion. Bowel injury (BI) is a well-recognized albeit uncommon complication. Guidelines on the insertion of SPC have been developed by the British Association of Urological Surgeons, but there remains little evidence regarding the incidence of this complication. This study uses contemporary UK data to assess the incidence of SPC insertion and the rate of BI and compares to a meta-analysis of available papers. National Hospital Episodes Statistics data were searched on all SPC insertions over an 18-month period for operating procedure codes, Code M38.2 (cystostomy and insertion of a suprapubic tube into bladder). Patients age, 30-day readmission rates, 30-day mortality rate, and catheter specific complication rate were collected. To estimate the BI rate, we searched patients who had undergone any laparotomy or bowel operation within 30 days of SPC insertion. Trusts were contacted directly and directed to ascertain whether there was SPC-related BI. PubMed search to identify papers reporting on SPC related BI was performed for meta-analysis RESULTS: 11 473 SPC insertions took place in the UK in this time period. One hundred forty-one cases had laparotomy within 30 days. Responses from 114 of these cases reported one BI related to SPC insertion. Meta-analysis showed an overall BI rate of 11/1490 (0.7%). This is the largest dataset reported on SPC insertions showing a lower than previously reported rate of BI. We recommend clinicians use a risk of BI of less than 0.25% when counseling low-risk patients.


Pharmacologic evidence that high dose chronic amitriptyline and desipramine down-regulate alpha 2-receptor-mediated hypothermia in the rat.
The effects of a three week course of treatment of amitriptyline (AMI) and desipramine (DMI) (3.2 and 10 mg/kg, s.c., b.i.d.) were studied on the hypothermic response to clonidine (0.1 mg/kg, s.c.) in male adult albino rats. Single doses of clonidine in normal unmedicated rats produced a marked (about 2 degrees C) fall in body temperature. In contrast, 0.9% NaCl control injections had no significant effect. The clonidine-induced hypothermia was prevented by yohimbine pretreatment (3.2 mg/kg, i.p., 1/2 hr before), suggesting an alpha 2-mediated response. On days 8 and 14 of chronic tricyclic antidepressant (TCA) treatment, the hypothermic response was prolonged. Four days after the abrupt withdrawal of 20 days of TCA treatment (10 mg/kg, s.c., b.i.d.), the clonidine-induced hypothermia response was attenuated consistent with down-regulation of alpha 2-receptors. This latter effect was not observed with a dose of 3.2 mg/kg, s.c., b.i.d. of either TCA.


Effects of ginsenoside Rg1 of Panax ginseng on mitosis in human blood lymphocytes in vitro.
The ginsenoside Rg1 extracted from the root of Panax ginseng can promote mitosis in cultured human lymphocytes activated by PHA or Con A. Its most effective concentrations are around 0.0003-0.0005 mg per ml of medium. Experiments show that it does not arrest the cells at any particular mitotic stage. It can also enhance the DNA synthesis in the activated lymphocytes. As a result of the increased number of the mitotic cells and enhanced DNA synthesis, the cell density is significantly increased in the Rg1-treated culture as compared with the control. However, in the absence of a mitogenic lectin Rg1 cannot restart the quinescent human lymphocytes to divide in vitro; therefore it is not mitogenic to resting cells. The possible action of Rg1 on activated human lymphocytes as well as its pharmacological significance are discussed.


Levels of polychlorinated biphenyls (PCBs) and three organochlorine pesticides in fish from the Aleutian Islands of Alaska.
Persistent organic pollutants (POPs), including polychlorinated biphenyls (PCBs) and chlorinated pesticides, have been shown to have many adverse human health effects. These contaminants therefore may pose a risk to Alaska Natives that follow a traditional diet high in marine mammals and fish, in which POPs bioaccumulate. This study examined the levels of PCBs and three pesticides [p, p'-DDE, mirex, and hexachlorobenzene (HCB)] in muscle tissue from nine fish species from several locations around the Aleutian Islands of Alaska. The highest median PCB level was found in rock sole (Lepidopsetta bilineata, 285 ppb, wet weight), while the lowest level was found in rock greenling (Hexagrammos lagocephalus, 104 ppb, wet weight). Lipid adjusted PCB values were also calculated and significant interspecies differences were found. Again, rock sole had the highest level (68,536 ppb, lipid weight). Concerning the PCB congener patterns, the more highly chlorinated congeners were most common as would be expected due to their greater persistence. Among the pesticides, p, p'-DDE generally dominated, and the highest level was found in sockeye salmon (Oncorhynchus nerka, 6.9 ppb, wet weight). The methodology developed by U.S. Environmental Protection Agency (USEPA) was used to calculate risk-based consumption limits for the analyzed fish species. For cancer health endpoints for PCBs, all species would trigger strict advisories of between two and six meals per year, depending upon species. For noncancer effects by PCBs, advisories of between seven and twenty-two meals per year were triggered. None of the pesticides triggered consumption limits. The fish analyzed, mainly from Adak, contain significant concentrations of POPs, in particular PCBs, which raises the question whether these fish are safe to eat, particularly for sensitive populations. However when assessing any risk of the traditional diet, one must also consider the many health and cultural benefits from eating fish.


The Effect of Botulinum Toxin A Injections in the Spine Muscles for Cerebral Palsy Scoliosis, Examined in a Prospective, Randomized Triple-blinded Study.
A prospective, randomized triple-blinded cross-over design treating with either botulinum toxin A (BXT) or saline (NaCl). To examine the efficacy of BTX treatment in cerebral palsy scoliosis (CPS). Intramuscular injections with BTX have been used off label in treating CPS. 1 prospective study has been conducted, demonstrating in both radiological and clinical improvement, whereas showing no side effects or complications. Subjects (brace-treated CPS between 2 and 18 yr) were injected using ultrasonic-guidance with either NaCl or BTX in selected spine muscles with 6 mo intervals (block randomization, sealed envelope). Radiographs of the spine and clinical follow-up were captured before and 6 weeks after each injection. Primary outcome parameter was radiological change in Cobb angle, where a 7° change was regarded as an effect (1 SD). Radiological parameters were measured before and 6 weeks after treatment by 3 experienced doctors separately. Moreover, clinical results were evaluated by the pediatric quality of life score and systematic open questioning of the parents about the child's wellbeing. Subjects, researchers, and monitors were blinded during the trial. Appropriate permissions (2008-004584-19) and no funding were obtained. 16 cerebral palsy patients (GFMCS III-V) with CPS were consecutively included, whereas 6 patients were excluded. There were no drop-outs to follow-up, but 1 possible serious adverse event of pneumonia resulting in death was recorded and the study was terminated. No significant radiological or clinical changes were detected when compared with NaCl injections using Wilcoxon matched pair signed-rank test. No positive radiological or clinical effects were demonstrated by this treatment, except for the parent's initial subjective but positive appraisal of the effect. However, the study was terminated due to 1 possible severe adverse event and scheduled numbers needed to treat (hence power) were not reached. 1.


Non-invasive measures of pulse wave velocity correlate with coronary arterial plaque load in humans.
Arterial stiffness is an emerging major risk factor for cardiovascular morbidity and mortality. The aim of the present study was to assess if coronary artery plaque load correlates with non-invasive measures of arterial stiffness. Prospective investigational study. Tertiary university hospital centre. Patients undergoing elective diagnostic coronary angiography. Coronary artery plaque burden was assessed using a 30 MHz intravascular ultrasound catheter during an automated pullback. Proximal coronary artery plaque volume was determined using a validated edge-detection algorithm following three-dimensional computerized reconstruction. Central arterial stiffness was assessed in each patient using applanation tonometry to radial, carotid and femoral pulses, with derivation of aortic pressure augmentation and pulse wave velocity using pulse wave analysis. In 35 patients (61 +/- 2 years), proximal coronary arterial plaque volume was 5.9 +/- 0.6 mm3/mm of vessel. Plaque volume correlated positively with carotid-radial pulse wave velocity (r = 0.47, P = 0.008) and appeared to correlate with carotid-femoral pulse wave velocity (r = 0.34, P = 0.07). Aortic augmentation (r = 0.24, P = 0.16), augmentation index (r = 0.3, P = 0.08), and pulse pressure (r = 0.22, P = 0.2) did not correlate significantly with proximal coronary artery plaque volume. Non-invasive measures of carotid-radial pulse wave velocity correlate with the extent of coronary artery plaque volume and may be a useful non-invasive surrogate marker for the extent of coronary atherosclerosis. Our findings are consistent with the suggestion that central aortic stiffness may promote the development of coronary atherosclerosis and ischaemic heart disease.


The design of the optic nerve in fish.
Fish have large eyes, with short optic nerves that are continually flexed by compensatory eye movements during swimming. Here, I review the tissue construction of the fish optic nerve, to see how the glia and axons are adapted to withstand these mechanical stresses, which are not normally encountered by CNS tissue within the skull. As in other lower vertebrates, the optic nerve astrocytes are highly unusual: their intermediate filaments are composed of cytokeratins (Giordano et al., 1989), not GFAP. Their processes are linked together by desmosomes, forming thin transverse lace-like partitions, placed at quasi-regular intervals longitudinally (Maggs & Scholes, 1990). This accordion-like arrangement is interpreted as providing a flexible tissue-skeleton for the optic nerve. A new observation is that the optic axons run in coherent parallel waves. This pattern, which is complementary to that of the astroglia, reversibly accommodates limited axial stretches. The waves are equivalent to those underlying the optical banding of Fontana (1781) in peripheral nerves, but wavelength (30 microns) and amplitude (5 microns) are about an order of magnitude less, reflecting the much smaller average size of the optic axons. The pattern also occurs in mammals, and may be restricted to the visual pathway: if present elsewhere in the CNS, nerve-fiber waves are inconspicuous at best. In fish, the astroglial partitions occur in register with the waves, suggesting that steric interactions between developing axons and glia may help to establish, or stabilize, the regular longitudinal spacing. This may have functional as well as mechanical implications, since the astrocytes form perinodal associations and their pattern is one which strongly clusters the nodes of Ranvier.


Effect of fertilizers on galanthamine and metabolite profiles in Narcissus bulbs by 1H NMR.
Narcissus bulbs contain the biologically active alkaloid galanthamine, and Narcissus is being developed as a natural source of the molecule for the pharmaceutical industry. The effect of fertilizer on galanthamine production was investigated in a field study using a (1)H nuclear magnetic resonance (NMR) metabolite profiling approach. Galanthamine was quantitated and major metabolites in the bulbs were identified. The application of standard fertilization levels of nitrogen and potassium caused a significant increase in galanthamine as compared to a control. Multivariate data analysis of the (1)H NMR data revealed that applying double the standard level of nitrogen fertilizer resulted in production of more amino acids and citric acid cycle intermediates, but not more galanthamine. The results indicated that standard levels of fertilizer currently applied in The Netherlands are sufficient for optimal galanthamine accumulation in the bulbs. This study shows how (1)H NMR-based metabolic profiling can provide insight into the response of plant metabolism to agricultural practices.


Motivational factors for initiating and maintaining physical activity among adults aged over fifty targeted by a tailored intervention.
Objective: To provide insight into the motivational working mechanisms (i.e. mediators) of an effective physical activity (PA) intervention for adults aged over fifty.Design: The mediation model (N = 822) was investigated in an RCT for the total intervention population, participants who were not norm-active at baseline (targeting PA initiation) and norm-active participants (targeting PA maintenance) separately.Main Outcome Measures: Potential mediators (attitude, self-efficacy, intention, action planning and coping planning) of the effect on PA (6-months) were assessed at baseline, 3 and/or 6 months.Results: The intervention resulted in a decrease in intention (B= -.209; p=.017), and an increase in action planning (B=.214; p=.018) and PA (B=.220; p=.002). Intention and action planning did not mediate the effect on PA. Self-efficacy, although not significantly influenced by the intervention, was found to be the only motivational variable that predicted change in PA (B=.164; p=.007). These results were confirmed among participants initiating PA. Among norm-active participants no significant intervention effects were identified.Conclusion: The motivational factors cannot explain the intervention effect on PA. Most likely, the effect can be explained by an interaction between the motivational factors together. Differences between participants initiating versus maintaining PA, highlight the importance of performing mediation analyses per subgroup.


A single-step sizing and radiofrequency ablation catheter for circumferential ablation of Barrett's esophagus: Results of a pilot study.
The 360 Express balloon catheter (360 Express) has the ability to self-adjust to the esophageal lumen, ensuring optimal tissue contact. The objective of this article is to evaluate the efficacy and safety of the 360 Express for radiofrequency ablation (RFA) treatment of Barrett's esophagus (BE). BE patients with low-grade dysplasia (LGD), high-grade dysplasia (HGD) or early cancer (EC) were included. Visible lesions were removed by endoscopic resection (ER) prior to RFA. RFA was performed with the 360 Express using the standard ablation regimen (12J/cm2-clean-12J/cm2). Primary outcome: BE regression percentage at three months. Secondary outcomes: procedure time, adverse events, complete eradication of dysplasia (CE-D) and intestinal metaplasia (CE-IM). Thirty patients (median BE C4M6) were included. Eight patients underwent ER prior to RFA. Median BE regression: 90%. Median procedure time: 31 minutes. Adverse events (13%): laceration (n = 1); atrial fibrillation (n = 1); vomiting and dysphagia (n = 1); dysregulated diabetes (n = 1). After subsequent treatment CE-D and CE-IM was achieved in 97% and 87%, respectively. In 10% a stenosis developed during additional treatment requiring a median of one dilation. This study shows that circumferential RFA using the 360 Express may shorten procedure time, while maintaining efficacy compared to standard circumferential RFA.


Exploring features of integrative teaching through a microanalysis of connection-making processes in a health sciences curriculum.
The interconnected nature of knowledge in the health sciences is not always reflected in how curricula, courses, and learning activities are designed. Thus have scholars advocated for more explicit attention to connection-making, or integration, in teaching and learning. However, conceptual and empirical work to guide such efforts is limited. This study analyzed classroom processes to determine what connections educators promoted in their classrooms and how those connections were made. A qualitative, focused ethnography design explored connection-making in a health science curriculum. Eight instructors were observed during ten class sessions resulting in 35 h of video data. Video data were entered into the observational software, Noldus Observer, and coded using continuous sampling. Frequency and duration of connections made were calculated in Noldus. Connection-making involved four interactive elements: The topic under direct consideration, other domains of professional knowledge (practice, student experience, research, theory, other content, core construct of the profession, external influences, metacognition), the integrative processes instructors used to connect a topic to other knowledge domains (informal example, stories, questioning, linking statements, formal cases, and program descriptions), and the learning and instructor context (type and purpose of course, instructor personal and professional experience). These elements are presented as an initial integrative learning taxonomy that can be used to guide explicit attention to connection-making in education and research.


ADIPOR1 deficiency-induced suppression of retinal ELOVL2 and docosahexaenoic acid levels during photoreceptor degeneration and visual loss.
Lipid metabolism-related gene mutations can cause retinitis pigmentosa, a currently untreatable blinding disease resulting from progressive neurodegeneration of the retina. Here, we demonstrated the influence of adiponectin receptor 1 (ADIPOR1) deficiency in retinal neurodegeneration using Adipor1 knockout (KO) mice. Adipor1 mRNA was observed to be expressed in photoreceptors, predominately within the photoreceptor inner segment (PIS), and increased after birth during the development of the photoreceptor outer segments (POSs) where photons are received by the visual pigment, rhodopsin. At 3 weeks of age, visual function impairment, specifically photoreceptor dysfunction, as recorded by electroretinography (ERG), was evident in homozygous, but not heterozygous, Adipor1 KO mice. However, although photoreceptor loss was evident at 3 weeks of age and progressed until 10 weeks, the level of visual dysfunction was already substantial by 3 weeks, after which it was retained until 10 weeks of age. The rhodopsin mRNA levels had already decreased at 3 weeks, suggesting that reduced rhodopsin may have contributed to early visual loss. Moreover, inflammation and oxidative stress were induced in homozygous KO retinas. Prior to observation of photoreceptor loss via optical microscopy, electron microscopy revealed that POSs were present; however, they were misaligned and their lipid composition, including docosahexaenoic acid (DHA), which is critical in forming POSs, was impaired in the retina. Importantly, the expression of Elovl2, an elongase of very long chain fatty acids expressed in the PIS, was significantly reduced, and lipogenic genes, which are induced under conditions of reduced endogenous DHA synthesis, were increased in homozygous KO mice. The causal relationship between ADIPOR1 deficiency and Elovl2 repression, together with upregulation of lipogenic genes, was confirmed in vitro. Therefore, ADIPOR1 in the retina appears to be indispensable for ELOVL2 induction, which is likely required to supply sufficient DHA for appropriate photoreceptor function and survival.


Endogenous brain oscillations and related networks detected by surface EEG-combined fMRI.
It is difficult to study the brain "at rest" with an approach generally pursued in science when external manipulation (independent variable) is used to obtain informative measurements (dependent variable) about the object of interest. External manipulation in its classic sense may suspend the resting state, and hence the object of interest will evade. Naturally, unless in a final and irreversible state, biological rest will always be an endogenously dynamic process. Combining two modalities, electroencephalography (EEG) and functional magnetic resonance imaging (fMRI), to simultaneously measure the brain's activity from two angles, one can be chosen to be interpreted as the independent variable and the other as the dependent variable, and without external manipulation the brain's spontaneous dynamics can be studied. The EEG, for example, observes endogenous modulations of vigilance and detects spontaneous events such as sleep spindles or epileptic discharges and can be used as the independent variable, i.e., to form a regressor to interrogate the fMRI data (dependent variable). The opposite is possible as well, and data fusion attempts try using all data both as dependent and independent variables at the same time. This review limits itself to an exemplary discussion of simultaneous EEG/fMRI studies in humans, and among a variety of proposed resting state networks only discusses a few, especially those for which non-resting state literature has proposed a functional meaning: the "default mode" network and an attentional network. It will be shown that one EEG feature can correlate with different fMRI activation maps and that a single resting state network may be associated with a variety of EEG patterns giving insight into the function of different resting states and the relationship between the two modalities in themselves.


Ranking of P-glycoprotein substrates and inhibitors by a calcein-AM fluorometry screening assay.
In order to compare the capacities of a variety of compounds to interfere with P-glycoprotein (Pgp) function, a novel assay was set up to work on a large screening scale. The model assay measures the capacity of parental sensitive (Par) and multidrug-resistant (MDR) cells to efflux a small fixed amount of acetoxymethyl calcein (calcein-AM) after their pretreatment with concentration ranges of known Pgp modulators. This microplate cytometry-based assay was performed with two different pairs of cell lines, the human lymphocytic leukemia CEM cells and the murine monocytic leukemia P388 cells. For a given Pgp-expressing MDR cell line, a Pgp modulator EC50 was defined as the concentration required to restore half of the calcein retention shown by similarly treated Par cells. With both MDR-P388 and MDR-CEM cells, EC50 comparisons ranked five reference Pgp modulators as follows: SDZ 280-446 > SDZ PSC 833 > cyclosporin A > verapamil > vinblastine. Further use of the MDR-CEM cells could rank 15 Pgp modulators for their capacity to interfere with calcein-AM efflux as follows: SDZ 280-446 1.9 x > SDZ PSC 833 8.3 x > cyclosporin A 3.8 x > amiodarone 1.1 x > quinacrine 1.6 x > verapamil 1.4 x > quinidine 1.1 x > vinblastine 11 x > vincristine 2 x > chloroquine > beta-lumicolchicine > or = gamma-lumicolchicine > or = colchicine > etoposide > or = doxorubicin. This calcein-AM assay should open the way for ranking large numbers of novel structures for their potential Pgp modulator properties, particularly for an efficient screening of Pgp function antagonists, but it does not allow defining whether their inhibition may be competitive or not.


Dental implant education in Nigeria: a curriculum guide.
Anecdotal evidence suggests that the practice of implant dentistry, "gold standard" for edentulism is presently very low in Nigeria. This article proposes a model for teaching dental implant education and also emphasises the need for inclusion of standardised dental implant education in the dental curriculum in Nigeria. A literature search using the MEDLINE was conducted on various models for teaching of dental implant education worldwide. Implant dental education for undergraduate and postgraduate programmes has 3 components namely: didactic educational programme, laboratory hands-on experience, and clinical hands-on experience. In addition, dentists and dental educationists can gain proficiency in basic and advanced implant dentistry through implant dental education organised by recognised universities, private and international implant organisations, manufacturer-sponsored educational programmes as well as training and courses organised by dental associations. Dental implant education should be given a priority and be included in both the undergraduate and postgraduate dental curriculum in Nigeria. In addition, a standardised implant dentistry programme should be formulated and developed for all dental schools in Nigeria.


Methylenetrahydrofolate Reductase Gene C677T Polymorphism and Diabetic Retinopathy: a Meta-Analysis.
Objective To investigate the association between the methylenetetrahydrofolate reductase gene C677T (MTHFR C677T) polymorphism and diabetic retinopathy (DR). Methods A total of 6971 subjects including 2707 DR patients and 4264 controls from 23 studies were enrolled in the study. A random-effects model was applied to estimate the overall effects and the stratified effects of the MTHFR C677T polymorphism on the risk of DR, and study quality was also assessed. Results Strong associations were observed between the MTHFR C677T polymorphism and DR. The carries of MTHFR C677T were more likely to be found in the DR group in relative to the healthy control group with odds ratio 1.68, 2.55, and 2.31 respectively in allele contrast model (T vs. C, 95%CI: 1.29-2.18, P<0.001, I 2=78.4%), homozygous model (TT vs. CC, 95%CI: 1.70-3.83, P=0.008, I 2=54.4%) and dominant model (TT+CT vs. CC, 95%CI: 1.62-3.29, P<0.001, I 2=74.7%). This association can also be found in contrast to the Ncd (non-complicated diabetic mellitus) group (allele contrast, OR=1.50, 95%CI: 1.07-2.11, P=0.032, I 2=62.1%; homozygous, OR=2.39, 95%CI: 1.06-5.38, P=0.017, I 2=66.7%; dominant, OR=1.59, 95%CI: 0.97-2.62, P=0.056, I 2=56.5%). For the heterozygous model (CT vs. CC), the association was significant in contrast to the healthy control group (OR=1.46, 95%CI: 1.64-3.69, P=0, I 2=77.3%), while in contrast to the Ncd control group the association was not statistically meaningful (OR=1.38, 95%CI: 0.87-2.18, P=0.131, I 2=43.7%). For the recessive model, 1.92-fold increased risk was found only in contrast to the Ncd control group (95%CI: 1.07-3.43, P=0.064, I 2=55.0%). There was no significant association found in the models in contrast to the DM control group. Conclusion In this meta-analysis, we found an association between the MTHFR C677T polymorphism and DR, especially in contrast to the Ncd control group. Further studies are required to establish more definite relationship.


E2F-1 lacking the transcriptional activity domain induces autophagy.
The transcription factor E2F-1 plays a crucial role in the control of cell proliferation. E2F-1 has tumor suppressive properties by inducing apoptosis and autophagy. In this study, E2F-1 and its truncated form (E2Ftr), lacking the transactivation domain (TAD), were compared for their ability to induce autophagy. In Gaussia luciferase-based assays, both E2F-1 and E2Ftr induced the proteolytic cleavage of the autophagic marker LC3. In addition, LC3 and autophagy protein 5 (Atg5) were upregulated by E2F-1 and E2Ftr. Likewise, both E2F proteins induced a punctate pattern of GFP-tagged LC3, indicating autophagosome formation. The presence of double-membrane autophagic vesicles induced by E2F-1 and E2Ftr was confirmed by transmission electron microscopy (TEM). The application of z-VAD-fmk, a caspase inhibitor, partially blocked both E2F-1 and E2Ftr-mediated cytotoxicity. Moreover, Atg5 (-/-) cells were more resistant to the E2F-1 or E2Ftr-induced cell killing effect than Atg5 wt cells. The TAD of E2F-1 is not essential for induction of autophagy; apoptosis and autophagy cooperate for an efficient cancer cell killing effect induced by E2F-1 or E2Ftr. E2Ftr-induced autophagy is a promising approach to destroy tumors that are resistant to conventional treatments.


Nanoconfined NaAlH4: prolific effects from increased surface area and pore volume.
Nanoconfinement is a promising technique to improve the properties of nanomaterials such as the kinetics for hydrogen release and uptake and the stability during cycling. Here we present a systematic study of nanoconfined NaAlH4 in nanoporous scaffolds with increasing surface area and pore volume and almost constant pore sizes in the range of 8 to 11 nm. A resorcinol formaldehyde carbon aerogel was CO2-activated under different conditions and provided aerogels with BET surface areas of 704, 1267 and 2246 m(2) g(-1) and total pore volumes of 0.91, 1.30 and 2.21 mL g(-1), respectively. Nanoconfinement of NaAlH4 was achieved by melt infiltration and (27)Al MAS NMR reveals that the respective scaffolds incorporate 68, 82 and 91 wt% NaAlH4, for the above-mentioned samples, while the remaining fraction decomposes to metallic Al indicating that increasing CO2-activation tends to facilitate the infiltration process. The frequencies for the (23)Na and (27)Al MAS NMR centerband resonances from NaAlH4 vary systematically for the infiltrated samples and are shifted towards higher frequency and become more narrow with increasing degree of CO2 activation of the scaffolds. This new effect is attributed to increasing interactions with conduction electrons from increasingly graphite-/graphene-like scaffolds. The bulk versus nanoconfined ratio of NaAlH4 was investigated using Rietveld refinement, revealing that the majority of added NaAlH4 is confined inside the nanopores. The hydrogen desorption kinetics decreased with increasing surface area and the hydrogen storage capacity is more stable and decreases less during continuous hydrogen release and uptake cycles. In fact, the available amount of hydrogen (2.7 wt% H2) was more than doubled compared to the nanoconfinement in the non-activated carbon aerogel (1.3 wt% H2). Furthermore, it was demonstrated that Ti-functionalization of the CO2-activated aerogels combines the high storage capacity with fast hydrogen release kinetics from NaAlH4 which fully decomposes into Na3AlH6 at T ≤ 100 °C.


Doping of graphene induced by boron/silicon substrate.
In this work, we show the doping of graphene most likely from heteroatoms induced by the substrate using Raman spectra, x-ray photoelectron spectroscopy, energy dispersive x-ray spectroscopy and ab initio molecular dynamics (MD) simulations. The doping of graphene on a highly boron-doped silicon substrate was achieved by an annealing at 400 K for about 3 h in an oven with air flow. With the same annealing, only the Raman features similar to that from the pristine graphene were observed in the freestanding graphene and the graphene on a typical Si/SiO2 wafer. Ab initio MD simulations were performed for defected graphene on boron-doped silicon substrate at several temperatures. All vacancy sites in the graphene are occupied either with B atoms or Si atoms resulting in the mixed boron-silicon doping of the graphene. The MD simulations validated the experimetal finding of graphene doped behavior observed by Raman spectrum. The electronic structure analysis indicated the p-type nature of doped graphene. The observed doping by the possible incorporation of heteroatoms into the graphene, simply only using 400 K annealing the boron-doped Si substrate, could provide a new approach to synthesize doped graphene in a more economic way.


Dynamic and opposing adjustment of movement cancellation and generation in an oculomotor countermanding task.
Adaptive adjustments of strategies help optimize behavior in a dynamic and uncertain world. Previous studies in the countermanding (or stop-signal) paradigm have detailed how reaction times (RTs) change with trial sequence, demonstrating adaptive control of movement generation. Comparatively little is known about the adaptive control of movement cancellation in the countermanding task, mainly because movement cancellation implies the absence of an outcome and estimates of movement cancellation require hundreds of trials. Here, we exploit a within-trial proxy of movement cancellation based on recordings of neck muscle activity while human subjects attempted to cancel large eye-head gaze shifts. On a subset of successfully cancelled trials where gaze remains stable, small head-only movements to the target are actively braked by a pulse of antagonist neck muscle activity. The timing of such antagonist muscle recruitment relative to the stop signal, termed the "antagonist latency," tended to decrease or increase after trials with or without a stop-signal, respectively. Over multiple time scales, fluctuations in the antagonist latency tended to be the mirror opposite of those occurring contemporaneously with RTs. These results provide new insights into the adaptive control of movement cancellation at an unprecedented resolution, suggesting it can be as prone to dynamic adjustment as movement generation. Adaptive control in the countermanding task appears to be governed by a dynamic balance between movement cancellation and generation: shifting the balance in favor of movement cancellation slows movement generation, whereas shifting the balance in favor of movement generation slows movement cancellation.


Factors associated with completing comprehensive traumatic brain injury evaluation.
This study examined factors associated with Iraq and Afghanistan Veterans following up with the Department of Veterans Affairs (VA) comprehensive traumatic brain injury (TBI) evaluation after a positive first-level VA TBI screen. Participants included 465 Iraq and Afghanistan Veterans at one VA Medical Center and its five affiliated community-based outpatient clinics, with a positive initial TBI screen between April 1, 2007 and June 1, 2010. We found that 75% of Veterans completed the comprehensive TBI evaluation. Women were three times less likely to complete the comprehensive TBI evaluation than men, and those who endorsed post-traumatic stress disorder avoidance symptoms were nearly two times less likely to complete the comprehensive TBI evaluation. In contrast, headaches, Hispanic ethnicity, and the season of the initial TBI screen (summer vs. winter) were positively associated with completing a comprehensive TBI evaluation. A substantial minority of Veterans who screen positive on the VA initial TBI screen fail to present for the comprehensive TBI evaluation. Addressing specific gender-related issues, avoidance, and the timing of referrals in the context of VA polytrauma programs may increase the likelihood that Veterans receive further assessment, education, and early intervention for TBI or other mental health problems to prevent chronic postdeployment disability.


Direct visualization of DNA baton pass between replication factors bound to PCNA.
In Eukarya and Archaea, the lagging strand synthesis is accomplished mainly by three key factors, DNA polymerase (Pol), flap endonuclease (FEN), and DNA ligase (Lig), in the DNA replication process. These three factors form important complexes with proliferating cell nuclear antigen (PCNA), thereby constructing a platform that enable each protein factor to act successively and smoothly on DNA. The structures of the Pol-PCNA-DNA and Lig-PCNA-DNA complexes alone have been visualized by single particle analysis. However, the FEN-PCNA-DNA complex structure remains unknown. In this report, we for the first time present this tertiary structure determined by single particle analysis. We also successfully visualized the structure of the FEN-Lig-PCNA-DNA complex, corresponding to a putative intermediate state between the removal of the DNA flap by FEN and the sealing of the nicked DNA by Lig. This structural study presents the direct visualization of the handing-over action, which proceeds between different replication factors on a single PCNA clamp bound to DNA. We detected a drastic conversion of the DNA from a bent form to a straight form, in addition to the dynamic motions of replication factors in the switching process.


Case Report and Genomic Analysis of Cefiderocol-Resistant Escherichia coli Clinical Isolates.
Cefiderocol is a novel siderophore cephalosporin with in vitro activity against multidrug-resistant (MDR), gram-negative bacteria and intrinsic structural stability to all classes of carbapenemases. We sought to identify gene variants that could affect the mechanism of action (MOA) of cefiderocol. We report a case of bacteremia in a liver transplant candidate with a strain of carbapenem-resistant Escherichia coli that was found to be resistant to cefiderocol despite no prior treatment with this antimicrobial agent. Using whole-genome sequencing, we characterized the genomic content of this E coli isolate and assessed for genetic variants between related strains that were found to be cefiderocol susceptible. We identified several variants in genes with the potential to affect the mechanism of action of cefiderocol. The cefiderocol resistance in the E coli isolate identified in this study is likely due to mutations in the cirA gene, an iron transporter gene.


Effect of replacing calcium salts of palm oil with camelina seed at 2 dietary ether extract levels on digestion, ruminal fermentation, and nutrient flow in a dual-flow continuous culture system.
Camelina is a drought- and salt-tolerant oil seed, which in total ether extract (EE) contains up to 74% polyunsaturated fatty acids. The objective of this study was to assess the effects of replacing calcium salts of palm oil (Megalac, Church & Dwight Co. Inc., Princeton, NJ) with camelina seed (CS) on ruminal fermentation, digestion, and flows of fatty acids (FA) and AA in a dual-flow continuous culture system when supplemented at 5 or 8% dietary EE. Diets were randomly assigned to 8 fermentors in a 2 × 2 factorial arrangement of treatments in a replicated 4 × 4 Latin square design, with four 10-d experimental periods consisting of 7 d for diet adaptation and 3 d for sample collection. Treatments were (1) calcium salts of palm oil supplementation at 5% EE (MEG5); (2) calcium salts of palm oil supplementation at 8% EE (MEG8); (3) 7.7% CS supplementation at 5% EE (CS5); and (4) 17.7% CS supplementation at 8% EE (CS8). Diets contained 55% orchardgrass hay, and fermentors were fed 72 g of dry matter/d. On d 8, 9, and 10 of each period, digesta effluent samples were taken for ruminal NH3, volatile fatty acids, nitrogen metabolism analysis, and long-chain FA and AA flows. Statistical analysis was performed using the MIXED procedure (SAS Institute Inc., Cary, NC). We detected an interaction between FA source and dietary EE level for acetate, where MEG8 had the greatest molar proportion of acetate. Molar proportions of propionate were greater and total volatile fatty acids were lower on CS diets. Supplementation of CS decreased overall ruminal nutrient true digestibility, but dietary EE level did not affect it. Diets containing CS had greater biohydrogenation of 18:2 and 18:3; however, biohydrogenation of 18:1 was greater in MEG diets. Additionally, CS diets had greater ruminal concentrations of trans-10/11 18:1 and cis-9,trans-11 conjugated linoleic acid. Dietary EE level at 8% negatively affected flows of NH3-N (g/d), nonammonia N, and bacterial N as well as the overall AA outflow. However, treatments had minor effects on individual ruminal AA digestibility. The shift from acetate to propionate observed on diets containing CS may be advantageous from an energetic standpoint. Moreover, CS diets had greater ruminal outflow of trans-10/11 18:1 and cis-9,trans-11 conjugated linoleic acid than MEG diets, suggesting a better FA profile available for postruminal absorption. However, dietary EE at 8% was deleterious to overall N metabolism and AA outflow, indicating that CS can be fed at 5% EE without compromising N metabolism.


Structural characterization of human and bovine lung surfactant protein D.
Human and bovine surfactant proteins D (SP-D) were purified from late amniotic fluid and bronchioalveolar lavage on the basis of its Ca(2+)-dependent affinity for maltose. The molecular mass of a trimeric subunit was determined by matrix-assisted laser desorption ionization MS to lie in the range 115-125 kDa for human SP-D and 110-123 kDa for bovine SP-D. A single polypeptide chain was determined at 37-41 and 36-40 kDa for the human and bovine species respectively. The major parts of the primary structures of both SP-D molecules were determined by a combination of MS and Edman degradation. The heterogeneity in SP-D was caused mainly by a high number of post-translational modifications in the collagen-like region. Proline and lysine residues were partly hydroxylated and lysine residues were further O-glycosylated with the disaccharide galactose-glucose. A partly occupied N-linked glycosylation site was characterized in human SP-D. The carbohydrate was determined as a complex type bi-antennary structure, with a small content of mono-antennary and tri-antennary structures. No sialic acid residues were present on the glycan, but some had an attached fucose and/or an N-acetylglucosamine residue linked to the core. Bovine SP-D was determined as having a similar structure.


Up-regulation of cytolytic functions of human Vδ2-γ T lymphocytes through engagement of ILT2 expressed by tumor target cells.
In humans, the majority of peripheral blood γδ T cells expresses Vγ9Vδ2 T-cell receptors (TCR) and recognize nonpeptidic phosphorylated antigens. In contrast, most tissue-derived γδ T cells, which are located mainly in spleen and epithelia, preferentially use Vδ1 or Vδ3 chains paired with diverse Vγ chains to form their TCR. Our knowledge about the antigenic specificity and costimulation requirements of human Vδ2(-) γδ T cells remains limited. In an attempt to address this important issue, we characterized the specificity of a monoclonal antibody (mAb 256), screened for its ability to specifically inhibit cytolytic responses of several human Vδ2(-) γδ T-cell clones against transformed B cells. We show that mAb 256 does not target a TCR ligand but blocks key interactions between non-TCR molecules on effector γδ T cells and ILT2 molecule, expressed by tumor targets. In line with the previously reported specificity of this NK receptor for classic and nonclassic major histocompatibility complex (MHC) class I molecules, blockade of MHC class I/ILT2 interactions using MHC class I- or ILT2-specific mAbs and ILT2-Fc molecules inhibited tumor-induced activation of Vγ8Vδ3 T-cell clones. Therefore, this study describes a new cytotoxic T lymphocyte activation pathway involving MHC class I engagement on γδ T cells.


[Densitometric analysis of crystalline lens nucleus in the choice of strategy of surgical treatment of cataracts].
Computer analysis of TV images of the eye was used for quantitative evaluation of optic density of the lens nucleus in patients with cataracts in need of surgical treatment. Indications to cataract phacoemulsification were determined using classification of the degree of the lens nucleus maturity. This classification is based on the maximal values of optic density of the anterior third of the nucleus, obtained by the lens densitometry, coefficient of light absorption by the lens nucleus, and tentative subjective classification of the lens nucleus maturity by a color scale. In initial cataract (first degree) the mean maximal optic density of the anterior third of the nucleus is 55.8 +/- 5.7 arb. units and coefficient of the lens nucleus light absorption is 0.93, in nuclear brown cataract (fourth degree) these values are 158.7 +/- 24.4 arb units and 1.81, respectively. Cataract with brown enlarged nucleus involving the cortical layers of the lens and high optic density of these layers, with lens nucleus light absorption coefficient more than 2.5 is a contraindication to phacoemulsification.


Biodegradable composite porous poly(dl-lactide-co-glycolide) scaffold supports mesenchymal stem cell differentiation and calcium phosphate deposition.
In recent decades, tissue engineering strategies have been proposed for the treatment of musculoskeletal diseases and bone fractures to overcome the limitations of the traditional surgical approaches based on allografts and autografts. In this work we report the development of a composite porous poly(dl-lactide-co-glycolide) scaffold suitable for bone regeneration. Scaffolds were produced by thermal sintering of porous microparticles. Next, in order to improve cell adhesion to the scaffold and subsequent proliferation, the scaffolds were coated with the osteoconductive biopolymers chitosan and sodium alginate, in a process that exploited electrostatic interactions between the positively charged biopolymers and the negatively charged PLGA scaffold. The resulting scaffolds were characterized in terms of porosity, degradation rate, mechanical properties, biocompatibility and suitability for bone regeneration. They were found to have an overall porosity of ∼85% and a degradation half time of ∼2 weeks, considered suitable to support de novo bone matrix deposition from mesenchymal stem cells. Histology confirmed the ability of the scaffold to sustain adipose-derived mesenchymal stem cell adhesion, infiltration, proliferation and osteo-differentiation. Histological staining of calcium and microanalysis confirmed the presence of calcium phosphate in the scaffold sections.


Linear space-variant optical processing of 1-D signals.
In the past, most optical data processing systems have been restricted to performing linear space-invariant operations. However, a wide class of interesting data processing operations require linear space-variant filtering. Three methods for performing linear space-variant processing of 1-D inputs are described. Experimental results obtained with all three systems are presented, and their relative advantages and disadvantages are discussed.


Dietary lipids modify receptor- and non-receptor-dependent components of alpha 1-adrenoceptor-mediated contraction.
Dietary lipid modulation of alpha-adrenoceptor (adrenergic receptor)- and non-adrenoceptor-mediated contractile properties of isolated rat abdominal aortic segments were assessed during the early developmental period. Rats were raised from conception to 90 days of age on semisynthetic diets containing various types and amounts of lipids. Aortic segments from three groups of rats fed high-fat diets (15% wt/wt) consisting of olive oil, corn oil, or lard as the sole lipid sources were compared with those from rats fed a low-fat control diet containing corn oil (5% wt/wt). alpha-Adrenoceptor activities were assessed by measuring the norepinephrine dose response of the tissue rings with and without partial inactivation of alpha-receptors by benextramine. alpha-Adrenoceptor sensitivity to norepinephrine increased, whereas receptor affinity decreased significantly in rats raised on high-fat diets. Qualitative features of dietary lipids influenced non-adrenoceptor-dependent aspects of vascular contractility. Diets rich in polyunsaturated fatty acids (high- and low-fat corn oil) raised the maximum response to norepinephrine and the contractile response to 60 mM potassium compared with more-saturated diets (olive oil and lard). These results demonstrate an effect of chronic feeding of high dietary fat on alpha-adrenoceptor-mediated contractility of abdominal aortic rings from young Sprague-Dawley rats. Qualitative features of dietary lipids also appear to modify receptor-independent parameters of the contractile response of the arterial tissue rings in these animals.


Neural Sensors: Learning Pixel Exposures for HDR Imaging and Video Compressive Sensing With Programmable Sensors.
Camera sensors rely on global or rolling shutter functions to expose an image. This fixed function approach severely limits the sensors' ability to capture high-dynamic-range (HDR) scenes and resolve high-speed dynamics. Spatially varying pixel exposures have been introduced as a powerful computational photography approach to optically encode irradiance on a sensor and computationally recover additional information of a scene, but existing approaches rely on heuristic coding schemes and bulky spatial light modulators to optically implement these exposure functions. Here, we introduce neural sensors as a methodology to optimize per-pixel shutter functions jointly with a differentiable image processing method, such as a neural network, in an end-to-end fashion. Moreover, we demonstrate how to leverage emerging programmable and re-configurable sensor-processors to implement the optimized exposure functions directly on the sensor. Our system takes specific limitations of the sensor into account to optimize physically feasible optical codes and we evaluate its performance for snapshot HDR and high-speed compressive imaging both in simulation and experimentally with real scenes.


Age-related improvements in auditory backward and simultaneous masking in 6- to 10-year-old children.
This study investigated the development of auditory frequency and temporal resolution using simultaneous and backward masking of a tone by a noise. The participants were 6- to 10-year-old children and adults. On the measure of frequency resolution (the difference in the detection threshold for a tone presented either in a bandpass noise or in a spectrally notched noise), 6-year-old children performed as well as adults. However, for the backward masking task, 6-year-olds had, on average, 34 dB higher thresholds than adults. A negative exponential decay function fitted to the backward masking data for subjects of all ages indicated that adult-like temporal resolution may not be reached until about 11 years of age. These results show that, measured by masking, frequency resolution has reached adult-like performance by 6 years of age, whereas temporal resolution develops beyond 10 years of age. Six-year-old children were also assessed with tests of cognitive ability. Improvements in both frequency and temporal resolution were found with increasing IQ score.


Lysine mutagenesis identifies cationic charges of human CYP17 that interact with cytochrome b5 to promote male sex-hormone biosynthesis.
Human CYP17 (17alpha-hydroxylase-17,20-lyase; also cytochrome P450c17 or cytochrome P450(17alpha)) catalyses a hydroxylation reaction and another reaction involving the cleavage of a C-C bond (the lyase activity) that is required only for androgen production. Single amino acid mutations in human CYP17, Arg(347)-->His and Arg(358)-->Gln, have been reported to result in the loss of the lyase activity and to cause sexual phenotypic changes in 46XY male patients. By using site-directed mutagenesis we show here that another mutation in human CYP17, Arg(449)-->Ala, for which human variants have yet not been described, also leads to selective lyase deficiency. Furthermore, all the three types of mutants display a loss of responsiveness to cytochrome b(5), an interaction that is essential for lyase activity, and hence male sex-hormone biosynthesis. That the defect could be essentially reversed by lysine mutagenesis has led to the conclusion that the cationic charges on all three residues (at the positions of Arg(347), Arg(358), Arg(449)) are vital for the functional interaction of CYP17 with cytochrome b(5) and that the loss of any one of these cationic charges is catastrophic.


Mussel defensins are synthesised and processed in granulocytes then released into the plasma after bacterial challenge.
MGD1 (Mytilus galloprovincialis defensin 1), a new member of the arthropod defensin family, is a 4 kDa antibacterial peptide previously isolated from the plasma of Mediterranean mussels. We report here the presence of MGD1 in the organelle-rich fraction of hemocytes and the cDNA sequence corresponding to MGD1 and one new isoform mRNA: MGD2. Sequence analysis indicated that MGDs are synthesised as precursors consisting of a putative signal peptide of 21 residues, the active peptide of 39 amino acids and a 21 residue carboxyl-terminal extension, rich in acidic amino acids. Localisation of the transcripts by northern blot revealed that the precursors are abundantly expressed in hemocytes. Immunocytochemistry at both the optical and ultrastructural levels showed that defensins (i) are predominantly located in vesicles of a granulocyte subclass of hemocytes containing small granules, (ii) are also found in large clear granules of another granulocyte subclass, and (iii) that MGD immune reactivity existed in granular structures of enterocytes. Finally, we revealed that bacterial challenge triggered a plasmatic increase of MGD1 concentration and gave evidence of the simultaneous release of the peptides from the hemocytes.


Sphingosine 1-phosphate protects bovine oocytes from heat shock during maturation.
Sphingosine 1-phosphate (S1P) is a sphingolipid metabolite that can block apoptosis by counteracting the proapoptotic effects of ceramide. Experiments were performed to evaluate whether S1P blocks the disruption in oocyte developmental competence caused by heat shock. Cumulus-oocyte complexes (COCs) were placed in maturation medium and cultured at 38.5 or 41 degrees C for the first 12 h of maturation. Incubation during the last 10 h of maturation, fertilization, and embryonic development were performed at 38.5 degrees C. Heat shock during the first 12 h of maturation reduced cleavage rate, the number of oocytes developing to the blastocyst stage, and the percentage of cleaved embryo that subsequently developed to blastocysts. Addition of 50 nM S1P to maturation medium had no effect on oocytes matured at 38.5 degrees C but blocked effects of thermal stress on cleavage and subsequent development. The blastocysts formed at Day 8 did not differ between S1P and control groups in caspase activity, total cell number, or percentage of cells that were apoptotic. Blocking endogenous generation of S1P by addition of 50 nM N1N-dimethylsphingosine, a sphingosine kinase inhibitor, reduced or tended to reduce cleavage rate and blastocyst development regardless of whether maturation of COCs was at 38.5 or 41 degrees C. Results demonstrate that S1P protects oocytes from a physiologically relevant heat shock and affects oocyte maturation even in the absence of heat shock. The S1P-treated oocytes that survived heat shock and became blastocysts had a normal developmental potential as determined by caspase activity, total cell number, and percentage of apoptotic cells. Thus, modulation of developmental competence of oocytes using S1P may be a useful approach for enhancing fertility in situations where developmental competence of oocytes is compromised.


Evolution of AANAT: expansion of the gene family in the cephalochordate amphioxus.
The arylalkylamine N-acetyltransferase (AANAT) family is divided into structurally distinct vertebrate and non-vertebrate groups. Expression of vertebrate AANATs is limited primarily to the pineal gland and retina, where it plays a role in controlling the circadian rhythm in melatonin synthesis. Based on the role melatonin plays in biological timing, AANAT has been given the moniker "the Timezyme". Non-vertebrate AANATs, which occur in fungi and protists, are thought to play a role in detoxification and are not known to be associated with a specific tissue. We have found that the amphioxus genome contains seven AANATs, all having non-vertebrate type features. This and the absence of AANATs from the genomes of Hemichordates and Urochordates support the view that a major transition in the evolution of the AANATs may have occurred at the onset of vertebrate evolution. Analysis of the expression pattern of the two most structurally divergent AANATs in Branchiostoma lanceolatum (bl) revealed that they are expressed early in development and also in the adult at low levels throughout the body, possibly associated with the neural tube. Expression is clearly not exclusively associated with the proposed analogs of the pineal gland and retina. blAANAT activity is influenced by environmental lighting, but light/dark differences do not persist under constant light or constant dark conditions, indicating they are not circadian in nature. bfAANAT alpha and bfAANAT delta' have unusually alkaline (> 9.0) optimal pH, more than two pH units higher than that of vertebrate AANATs. The substrate selectivity profiles of bfAANAT alpha and delta' are relatively broad, including alkylamines, arylalkylamines and diamines, in contrast to vertebrate forms, which selectively acetylate serotonin and other arylalkylamines. Based on these features, it appears that amphioxus AANATs could play several roles, including detoxification and biogenic amine inactivation. The presence of seven AANATs in amphioxus genome supports the view that arylalkylamine and polyamine acetylation is important to the biology of this organism and that these genes evolved in response to specific pressures related to requirements for amine acetylation.


Carpal tunnel syndrome in patients on long-term hemodialysis.
Sixteen cases with carpal tunnel syndrome (CTS) were reported out of 412 patients on long-term hemodialysis due to chronic renal failure. Clinical symptoms included numbness in the area innervated by the median nerve in 100% of symptomatic hands and pain in the wrist and hand in 81%. This pain usually became severer at night and during hemodialysis. Muscle atrophy was noted in 41% of the hands. Nerve conduction studies revealed prolonged distal sensory latency, slowed sensory nerve conduction velocity across the wrist and normal distal motor latency in 18%, prolonged distal motor latency in 51% and no response in motor or sensory stimulation in 31%. Evidence of denervation on electromyography was seen in 36% of the hands. Patients were conservatively treated avoiding daily activities precipitating the condition with volar wrist splint only at night in 18% and in 90% with steroid hormone injection in the carpal tunnel. Median nerve release was performed in 18% of the hands. Amyloid deposit was demonstrated in 3 of 4 operated hands. Although the relation between long-term hemodialysis and the occurrence of the amyloid deposition in the carpal tunnel has not been established, the present data along with other recent reports strongly indicate that amyloid deposit in the carpal tunnel on hemodialysis patients could be one of the most possible cause of CTS.


Homocysteine-lowering therapy or antioxidant therapy for bone loss in Parkinson's disease.
We investigated whether homocysteine (Hcy)- lowering therapy or an antioxidant prevented bone loss in Parkinson's disease (PD) patients taking levodopa. Forty-two PD patients with low bone mineral density (BMD) taking levodopa were randomly assigned to Hcy-lowering therapy (5 mg folate and 1500 microg vitamin B(12) daily), alpha-lipoic acid (alpha-LA) therapy (1200 mg daily), or control groups. Primary outcomes were BMD changes from baseline to 12 months. Secondary outcomes were changes in Hcy level, and C-telopeptide (CTX) levels at 12 months. Forty-one patients completed the study. Hcy-lowering therapy resulted in significantly greater BMD changes at the lumbar spine (4.4%), total femur (2.8%), and femur shaft (2.8%) than control (P = 0.005-0.023). BMD changes in the alpha-LA therapy group were similar to those of the control group, but changes at the trochanter (4.6%) were significantly greater in the alpha-LA therapy group than in the control group after adjustment for body mass index changes. Hcy concentrations decreased to 35.2% +/- 13.4% in the Hcy-lowering therapy group, but increased in other groups. Serum CTX levels at 12 months tended to be lower in the Hcy-lowering group (0.442 +/- 0.024 ng/mL) than control group (0.628 +/- 0.039 ng/mL) (P = 0.159). This small trial suggests that Hcy-lowering therapy may prevent bone loss in PD patients taking levodopa.


Larva migrans in the oral mucosa: report of two cases.
Cutaneous Larva migrans is a very common disease in tropical regions. In the oral mucosa, the infection occurs in the same way as in the skin, but it is rarer. This report describes two cases of Larva migrans in the oral mucosa. The first case was in a 27-year-old woman who presented an erythematous plaque located on the buccal mucosa, extending to a posterior direction, following a linear pattern, to other areas of the mouth. After incisional biopsy of the anterior-most portion of the lesion, morphological details obtained in multiple examined sections suggested Necator or Ancylostoma braziliense larvae as the cause of infection. The second case was in a 35-year-old male who presented a fusiform erythematous plaque in the palatal mucosa. This area was removed and submitted to microscopic examination under a presumptive diagnosis of "parasite migratory stomatitis". The histological characteristics were suggestive of a larva pathway. In both cases the lesion disappeared after biopsy and the patients were symptom-free.


A descriptive epidemiology of giardiasis in New Zealand and gaps in surveillance data.
Giardia is the most commonly notified waterborne disease in New Zealand, which has high incidence rates compared with other developed countries. Four years of giardiasis notification data were analysed to describe the epidemiological patterns of infection in New Zealand and compared with local and international data. Anonymised information was collected nationally for 7818 notified cases of giardiasis between July 1996 and June 2000. International data were collected from the data sources of respective countries. Infection rates adjusted for confounding factors were calculated and presented by age, gender, ethnicity, and area using statistical and spatial methods. Most cases occurred in the 1-4 year age group followed by the 25-44 year age group, and were of Pakeha/European ethnicity. Ethnicity was unknown for 18% of cases, thus affecting demographic calculations. Rates were elevated for several Health Districts in New Zealand (West Coast, Wellington, Waikato, Auckland, Hawke's Bay, Hutt, Rotorua, and Tauranga). The higher rates of giardiasis observed in New Zealand, in comparison with other developed countries, may be related to environmental or social factors. Time-trend analysis suggests a seasonal pattern. This study identified vulnerable groups and major data-gaps. Recommendations for improvements in disease surveillance and data quality are discussed. Geographical information system (GIS) applications are useful for disease monitoring.