Gene-based approaches in pain research and exploration of new therapeutic targets and strategies.
Large panel of gene-based techniques is used for many years specifically in the pain research field. From the first identification (cloning) of some "mythic" genes, such as those encoding opioid or capsaicin receptors allowing then the creation of first-generation knockout mice, to the today conditional (time, tissue, cell-type and even pathology-dependent) and regulatable modulation of a gene function, these approaches largely contributed to fundamental leaps forward in our understanding of the function of some proteins and of their interest as possible druggable targets. Perhaps one of the most remarkable evolution in the last years is the passage of these approaches from the bench to the patient; whether it concerns the identification of genes involved in inherited pain insensibility/susceptibility, the search for genetic markers of pain types, the individual pharmacogenomics or even the first gene therapy trials. From many possible variants of gene-grounded techniques used in pain research we focus here on gene knockouts and some recent developments, on viral vectors-based gene transfer and on transgenic models for the tracing of pain pathways. Through these selected examples we attempted to emphasize the immense potential of these approaches and their already well-recognized contribution in both the basic and clinical pain research.


Evaluation of 2,6-diamino-N-([1-(1-oxotridecyl)-2-piperidinyl]methyl)- hexanamide (NPC 15437), a protein kinase C inhibitor, as a modulator of P-glycoprotein-mediated resistance in vitro.
We assessed the effect of the protein kinase C inhibitor 2,6-diamino-N-([1-(1-oxotridecyl)-2-piperidinyl]methyl)hexanami de (NPC 15437) on the action of anthracyclines, epipodophyllotoxins and vinca alkaloids in P-glycoprotein (Pgp)-expressing CH(R)C5 hamster ovary and MCF-7/Adria(R) human breast cancer cells. Flow microfluorimetry revealed that treatment of CH(R)C5 cells with 75 microM NPC 15437 for 1 h resulted in a 6- to 10-fold increase in the nuclear accumulation of daunorubicin. Colony forming assays revealed that treatment with 75 microM NPC 15437 was associated with a 4-fold decrease in the LD90 for etoposide and a 2.5-fold decrease in the LD50 for vincristine. At higher concentrations of NPC 15437, greater modulation of anthracycline accumulation was observed; but NPC 15437 itself inhibited subsequent colony formation. Similar effects on drug accumulation and cytotoxicity were observed in MCF-7/Adria(R) cells. Experiments designed to investigate the mechanism by which NPC 15437 exerts these effects revealed that treatment with the protein kinase C activator phorbol-12-myristate 12-acetate partially reversed the effect of NPC 15437, suggesting that NPC 15437 was exerting an effect through protein kinase C. Photoaffinity labeling experiments revealed that NPC 15437 also inhibited the binding of [3H]-azidopine to Pgp in isolated membrane vesicles. These results identify NPC 15437 [correction of NPC15437] as the prototype of a new class of potential Pgp modulators but indicate that the effects of this agent as a modulator are potentially limited by its cytotoxicity.


Foxg1 is required to limit the formation of ciliary margin tissue and Wnt/β-catenin signalling in the developing nasal retina of the mouse.
The ciliary margin (CM) develops in the peripheral retina and gives rise to the iris and the ciliary body. The Wnt/β-catenin signalling pathway has been implicated in ciliary margin development. Here, we tested the hypothesis that in the developing mouse retina Foxg1 is responsible for suppressing the Wnt/β-catenin pathway and restricting CM development. We showed that there is excess CM tissue in Foxg1(-/-) null embryos and this expansion is more pronounced in the nasal retina where Foxg1 normally shows its highest expression levels. Results on expression of a reporter allele for Wnt/β-catenin signalling and of Lef1, a target of Wnt/β-catenin signalling, displayed significant upregulation of this pathway in Foxg1(-/-) nulls at embryonic days 12.5 and 14.5. Interestingly, this upregulation was observed specifically in the nasal retina, where normally very few Wnt-responsive cells are observed. These results indicate a suppressive role of Foxg1 on this signalling pathway. Our results reveal a new role of Foxg1 in limiting CM development in the nasal peripheral retina and add a new molecular player in the developmental network involved in CM specification.


Menopause and development of Alzheimer's disease: Roles of neural glucose metabolism and Wnt signaling.
Late onset Alzheimer´s disease (AD) is a neurodegenerative disease with gender differences in its onset and progression, being the prevalence predominant in women and at an earlier age than in men. The pathophysiology of the menopausal condition has been associated to this dementia, playing major roles regarding both endocrine and glucose metabolism changes, amongst other mechanisms. In the current review we address the role of estrogen deficiency in the processes involved in the development of AD, including amyloid precursor protein (APP) processing to form senile plaques, Tau phosphorylation forming neurofibrillary tangles, Wnt signaling and AD neuropathology, the role of glucose brain metabolism, Wnt signaling and glucose transport in the brain, and our research contribution to these topics.


Intramuscular tetanus neurotoxin reverses muscle atrophy: a randomized controlled trial in dogs with spinal cord injury.
Motor symptoms of spinal cord injury (SCI) considerably impair quality of life and are associated with a high risk of secondary diseases. So far, no pharmacological treatment is available for these symptoms. Therefore, we conducted a randomized, double-blinded, placebo-controlled study in dogs with spontaneous SCI due to disc herniation to test whether a reduction of spinal inhibitory activity by intramuscular injections of tetanus neurotoxin (TeNT) alleviates motor symptoms such as muscle atrophy or gait function. To this end, 25 dogs were treated with injections of either TeNT or placebo into their paretic hindlimb muscles. Effects of TeNT on muscle thickness were assessed by ultrasound, while effects on gait function were measured using the modified functional scoring system in dogs. Four weeks after the TeNT injections, muscle thickness of the gluteus medius muscle (before median 1.56 cm [inter-quartile range {IQR} 1.34-1.71 cm] and after median 1.56 cm [IQR 1.37-1.85 cm], P-value 0.0133) as well as of the rectus femoris muscle (before median 0.76 cm [IQR 0.60-0.98 cm] and after median 0.93 cm [IQR 0.65-1.05 cm], P-value 0.0033) significantly increased in the TeNT group. However, there was no difference in gait function between the TeNT and placebo groups. The treatment was well tolerated by all dogs without any signs of generalized tetanus symptoms or any spreading of effects beyond the lumbar level of the injected hindlimbs. With regard to the beneficial effects on muscle thickness, intramuscular injections of TeNT represent the first pharmacological approach that focally reverses muscle atrophy in SCI. Moreover, the study data support the safety of this treatment when TeNT is used at low dose.


Developing quantitative physiological phenotypes of sleep apnea for epidemiological studies.
Existing physiological databases have not been sufficiently detailed to provide relevant and important information for characterizing the pathophysiology of obstructive sleep apnea. Critical collapsing pressure (P(CRIT)) is a standard method for determining upper airway patency during sleep, however is labor intensive and prohibits large-scale studies. Based on previously published data indicating R(US) does not significantly vary between groups, our aim was to develop an approach to estimate the P(CRIT) from airflow at atmospheric pressure (V(atm)). In a dataset of 126 subjects, where P(CRIT) and R(US) were measured using standard techniques. We then determined the minimum sample size required to estimate the R(US) mean and variance by utilizing a bootstrap procedure (30 times for n=3 to 126). We first estimated the minimum number of subjects needed for obtaining a group for a two-tailed (z=1.96) standard error for R(US) in the population. Then in 75 individuals, quantitative estimates of airflow were obtained at atmospheric pressure. Using the estimated R(US) and atmospheric, we determined an estimated P(CRIT) (ЄP(CRIT)). Bland-Altman plots were generated to determine the agreement between the measured P(CRIT) and ЄP(CRIT). For the entire population the mean ± SEM R(US) was 23 ± 1 cmH(2)O/L/s (± 95% CI: 21, 25). ~40 subjects represent the minimum sample required to estimate the population variance within ± 2 SEM. In the subsample with atmospheric flow measurements, a linear regression model (ЄP(CRIT) [cmH(2)O] = V(@PN) [L/s]x-23[cmH(2)O/L/s]), ЄP(CRIT) ranged from 0 to -9.6 cmH(2)O. In the Bland-Altman analysis there was no mean difference between the measured P(CRIT) and ЄP(CRIT) (-0.01 cmH(2)O; p=0.8) with upper and lower limits of agreement at ± 2.3 cmH(2)O. The variance of upstream resistance approaches a constant value in groups with approximately 40 subjects. Utilizing a fixed up-stream resistance to estimate P(CRIT) from the airflow at atmospheric pressure agrees with the measured values. These data suggest that measurements of quantitative airflow during standard polysomnography can be used to determine upper airway properties in large cohorts.


Relaxin influences the growth of MCF-7 breast cancer cells. Mitogenic and antimitogenic action depends on peptide concentration.
The effects of relaxin (RLX) on the human breast adenocarcinoma cell line MCF-7 have been evaluated. The cells were maintained in culture with Dulbecco's modified Eagle's medium with 1% and 10% fetal calf serum. Highly purified porcine RLX was added at concentrations ranging between 10(-11) and 10(-6) M, and, after 96-hour incubation in the presence of the peptide, cell proliferation, intracellular cyclic adenosine monophosphate (cAMP) content, percent cycling cells, and structural and ultrastructural pattern were studied. The results indicate that RLX is a direct modulator of MCF-7 cell proliferation, stimulating growth at low concentrations and inhibiting growth at high concentrations. Determinations of percent cycling cells and intracellular cAMP accumulation agree with the results of the growth studies. Addition of different concentrations of 8-Br-cAMP to the culture medium results in a dose-related stimulation of MCF-7 cell proliferation. Morphologic examination shows that, in the current experiments, RLX does not induce any clear-cut signs of differentiation of MCF-7 cells in terms of activation of secretion or intracellular lipid deposition. These findings indicate that RLX should be regarded as a novel agent involved in the control of growth of human breast cancer cells.


Inference from clinical and fluid dynamic studies about underlying cause of spontaneous isolated superior mesenteric artery dissection.
Due to increased use and improvements in diagnostic imaging studies, spontaneous isolated superior mesenteric artery dissection (SISMAD), which is a rare vascular event, has been reported to occur on a more frequent basis. Although there have been some anecdotal case reports describing the underlying pathology of SISMAD, the etiology of the majority of SISMAD is still poorly understood. The purpose of this study was to determine the underlying cause of SISMAD. From July 2001 to March 2010, 51 consecutive patients with SISMAD (symptomatic 39, asymptomatic 12) and 38 patients with combined aortic and superior mesenteric artery dissection (CASMAD) were identified in a single institution by retrospective investigations. Diagnosis was dependent on multi-detector helical computed tomography (CT) scan. To find clinical characteristics of SISMAD, we compared demographic, clinical, and lesion (site of entry tear, type, length) characteristics between the two groups. To find any flow dynamic abnormalities at the proximal segment of the superior mesenteric artery (SMA), we conducted flow dynamic studies using computational fluid dynamic models (V.12; ANSYS, Inc., Canonsburg, Pa). Streamline patterns and wall shear stress distributions were tested with computer simulation models using three different branching angles of SMA from the abdominal aorta. Compared to CASMAD, SISMAD was more common in men (90.2% vs 71.1%; P = .02), less frequently associated with hypertension (31.4% vs 65.8%; P = .001), and more frequently associated with intra-abdominal cancers (11.8% vs 0%; P = .036). In a fluid dynamic study using computational fluid dynamic models, we found abnormal mechanical stresses at the anterior wall around the convex portion of the SMA. Development of SISMAD seems to be less likely the result of hypertension or connective tissue disease but more likely due to hemodynamic force caused by convex curvature.


Novel 1,4-dihydropyridine (Bay K 8644) facilitates calcium-dependent [3H]noradrenaline release from PC 12 cells.
The effects of the novel 1,4-dihydropyridine Bay K 8644 [methyl-1,4-dihydro-2,6-dimethyl-3-nitro-4-(2-trifluoromethylphenyl)-pyridine- 5-carboxylate] on the release of [3H]noradrenaline in cultured PC 12 cells were investigated. K+ in a concentration-dependent manner evoked 3H-transmitter release with an EC50 of 50-56 mM. Bay K 8644 at 30 nM potentiated the K+-evoked [3H]noradrenaline release; however, in the absence of calcium neither K+ evoked nor Bay K 8644 enhanced [3H]noradrenaline release. At a K+ concentration of 25 mM, Bay K 8644 stimulated [3H]noradrenaline release fivefold, with an EC50 of 10 nM, and 100 nM of the calcium channel blocker nitrendipine shifted the concentration response curve of Bay K 8644 to the right in an apparently competitive fashion. Nitrendipine blocked the Bay K 8644-potentiated release with an EC50 of 700 nM in the presence of 500 nM Bay K 8644. [3H]Nitrendipine bound to a saturable population of binding sites on PC 12 cell membranes with a Bmax of 180 fmol X mg-1 of membrane protein and a KD of 0.9 nM. Bay K 8644 inhibited [3H]nitrendipine binding with a Ki of 16 nM. It is concluded that Bay K 8644 binds to, and stabilizes, the open state of calcium channels and thus acts as a "calcium agonist" to mediate calcium-dependent cellular events such as catecholamine release from PC 12 cells.


The effects of 1,25(OH)2 D3 treatment on immune responses and intracellular metabolic pathways of bone marrow-derived dendritic cells from lean and obese mice.
Vitamin D affects differentiation, maturation, and activation of dendritic cells (DCs). Obesity-related immune dysfunction is associated with metabolic changes in immune cells. Objectives of the study are to investigate the effects of vitamin D and obesity on immune responses and markers related to immunometabolism of bone marrow-derived dendritic cells (BMDCs). Bone marrow cells (BMCs) were isolated from lean and obese mice, and BMDCs were generated by culturing BMCs with rmGM-CSF. BMDCs were treated with 1 or 10 nM of 1,25-dihydroxyvitamin D3 (1,25(OH)2 D3 ), and maturation was induced by LPS (50 ng/ml) stimulation for 24 hr. Cell phenotypes, cytokine productions, and expression of proteins and genes involved in Akt/mTOR signaling pathway and glycolytic pathway were determined. 1,25(OH)2 D3 treatment inhibited differentiation of BMDCs (CD11c+ %), expression of phenotypes related with DC function (MHC class II and CD86) and production of IL-12p70 in both lean and obese mice. The expression of PD-L1 and the ratio of IL-10/IL-12p70 were increased by 1,25(OH)2 D3 . With 1,25(OH)2 D3 treatment, Akt/mTOR signaling pathway was suppressed, and expression of genes related to glycolysis (Glut1, Pfkfb4, and Hif1A) was increased. The upregulation of glycolysis-related genes observed with 1,25(OH)2 D3 treatment seems to be associated with the induction of tolerogenic features of BMDCs from lean and obese mice, and Hif1A seems to have a potential role in conveying the effect of 1,25(OH)2 D3 on glycolysis.


Influence of presentation at the weekend on treatment and outcome in ST-elevation myocardial infarction in hospitals with catheterization laboratories.
Studies about the influence of various factors on clinical therapy and course in acute coronary syndromes have shown that the outcome is related to admission time to the hospital, with an impaired prognosis in patients admitted out of regular working hours. However little is known about the impact of admission on weekend in hospitals with catheterisation laboratories. We analyzed data of the prospective MITRA-PLUS registry of 11,516 patients with ST-elevation myocardial infarction (STEMI) admitted to hospitals with catheterization facilities for differences of in-hospital mortality between patients admitted during regular working hours, at night and on weekends. The prehospital delay and "door-to-balloon"-time were significantly longer on weekends and at nights than at regular working hours (median 196 Vs. 240 Vs. 155 min; P < 0.0001; 60 Vs. 84 min at weekends, resp. 75 min at nights; P < 0.0001). Reperfusion therapy was performed in 72.8% (8,248/11,332) patients, and there were less patients treated on weekend versus "on"-hours (69.7 Vs. 77 %, P < 0.0001). On weekends we found a significant higher in-hospital mortality (11.1 Vs. 9.4%, P = 0.01) and at night there was a trend to higher in-hospital mortality when compared with regular working hours (10.6 Vs. 9.4%, P = 0.07). In patients with STEMI admitted to hospitals with catheterization facilities, admission during the "off"-hours is associated with higher in-hospital mortality. This may be due to lower rates of revascularization therapy and longer prehospital and in-hospital delays as compared to "on"-hours.


Diagnostic value of the skin-prick test and RAST assay in insect sting allergy.
The results of skin-prick tests to four concentrations of venom (0.1, 1, 10 and 1000 micrograms/ml) carried out on two occasions were analysed in relation to the history of adverse reactions to stings and to the level of venom-specific IgE antibody in serum, in forty-two subjects allergic to insect stings (sixteen to bee and twenty-six to wasp). Fifty control subjects (some of whom had never been stung by bee or wasp) with no history of adverse reaction to stings were also studied. No subject gave a positive skin-test reaction to 0.1 microgram/ml, and small numbers reacted to either 1 or 10 micrograms/ml. The lowest concentration of venom to which most subjects had a positive skin test was 100 micrograms/ml. Our data suggest that in wasp-allergic patients a positive skin test to 100 micrograms/ml is normally significant (reflecting the presence of specific IgE), whereas in bee-allergic patients a skin test reaction to 100 micrograms/ml is usually non-specific for the following reasons. (i) In the allergic patients when skin tests were repeated, a reaction to 100 micrograms/ml bee venom often became negative (in six of eight), whereas for wasp venom the reaction became more positive (at 10 or 1 micrograms/ml) in seven of eight patients. Whilst this might reflect lack of reproducibility, the consistent direction of change for either bee or wasp venom suggests responses to this concentration of these venoms may have different interpretations.(ABSTRACT TRUNCATED AT 250 WORDS)


Evaluation of Potential Drug-Drug Interaction Risk of Pexidartinib With Substrates of Cytochrome P450 and P-Glycoprotein.
Pexidartinib is approved for treatment of adults with symptomatic tenosynovial giant cell tumor. In vitro data showed pexidartinib's potential to inhibit and induce cytochrome P450 (CYP) 3A, inhibit CYP2C9, CYP2C19 and P-glycoprotein (P-gp). Herein, 2 open-label, single-sequence, crossover studies evaluated the drug-drug interaction potential of pexidartinib on CYP enzymes (CYP2C9, CYP2C19, and CYP3A) and P-gp. Thirty-two subjects received single oral doses of midazolam (CYP3A substrate) and tolbutamide (CYP2C9 substrate) alone and after single and multiple oral doses of pexidartinib. Twenty subjects received single oral doses of omeprazole (CYP2C19 substrate) and digoxin (P-gp substrate) alone or with pexidartinib. Analysis of variance was conducted to determine the effect of pexidartinib on various substrates' pharmacokinetics. No drug-drug interaction was concluded if the 90% confidence interval of the ratio of test to reference was within the range 80% to 125%. Coadministration of single and multiple doses of pexidartinib resulted in 21% and 52% decreases, respectively, in the area under the plasma concentration-time curve from time zero to the last measurable time point (AUClast ) of midazolam, whereas AUClast values of tolbutamide increased 15% and 36%, respectively. Omeprazole exposure decreased on concurrent administration with pexidartinib, the metabolite-to-parent ratio was similar following omeprazole administration alone vs coadministration with pexidartinib; pexidartinib did not affect CYP2C19-mediated metabolism. Maximum plasma concentrations of digoxin slightly increased (32%) with pexidartinib coadministration; no significant effect on digoxin AUClast . These results indicate that pexidartinib is a moderate inducer of CYP3A and a weak inhibitor of CYP2C9 and does not significantly affect CYP2C19-mediated metabolism or P-gp transport.


EGFR and myosin II inhibitors cooperate to suppress EGFR-T790M-mutant NSCLC cells.
An acquired mutation (T790M) in the epidermal growth factor receptor (EGFR) accounts for half of all relapses in non-small cell lung cancer (NSCLC) patients who initially respond to EGFR kinase inhibitors. In this study, we demonstrated for the first time that EGFR-T790M interacts with the cytoskeletal components, myosin heavy chain 9 (MYH9) and β-actin, in the nucleus of H1975 cells carrying the T790M-mutant EGFR. The interactions of EGFR with MYH9 and β-actin were reduced in the presence of blebbistatin, a specific inhibitor for the MYH9-β-actin interaction, suggesting that the EGFR interaction with MYH9 and β-actin is affected by the integrity of the cytoskeleton. These physical interactions among MYH9, β-actin, and EGFR were also impaired by CL-387,785, a kinase inhibitor for EGFR-T790M. Furthermore, CL-387,785 and blebbistatin interacted in a synergistic fashion to suppress cell proliferation and induce apoptosis in H1975 cells. The combination of CL-387,785 and blebbistatin enhanced the down-regulation of cyclooxygenase-2 (COX-2), a transcriptional target of nuclear EGFR. Overall, our findings demonstrate that disrupting EGFR interactions with the cytoskeletal components enhanced the anti-cancer effects of CL-387,785 against H1975 cells, suggesting a novel therapeutic approach for NSCLC cells that express the drug-resistant EGFR-T790M.


Does the lack of the P-glycoprotein efflux pump in neutrophils explain the efficacy of colchicine in familial Mediterranean fever and other inflammatory diseases?
Colchicine is an alkaloid drug commonly used in familial Mediterranean fever (FMF), gout, Behcet's syndrome, psoriasis and Sweet's syndrome. The exact mechanism of its action in these diseases is not entirely known. However, it has been shown that colchicine may inhibit neutrophil chemotaxis, thereby decreasing the inflammatory process. Recently, it was shown that colchicine accumulates in neutrophils in higher concentrations than in lymphomonocytes. Studies dealing with the multiple drug resistance (MDR) issue disclosed that neutrophils lack the P-glycoprotein (P-gly) membranal pump (encoded by the MDR1 gene). We propose that the preferential accumulation of colchicine in neutrophils compared with lymphomonocytes is due to the absence of the P-gly efflux pump in the former. This may explain the effectiveness of colchicine in diseases where increased chemotaxis is evident. The hypothesis may also provide an explanation for FMF patients who do not respond to the drug.


Tissue expressions of nine genes important to immune defence of the Pacific white shrimp Litopenaeus vannamei.
The tissue expressions of nine immune related genes in apparently healthy Pacific white shrimp Litopenaeus vannamei were analyzed by conventional RT-PCR, quantitative real time PCR (qPCR) and in situ hybridisation. The nine genes were beta-glucan binding protein-high density lipoprotein (BGBP-HDL), lipopolysaccharide-beta-glucan binding protein (LGBP), haemocyanin, prophenoloxidase (proPO), transglutaminase (TGase), crustins, penaeidins (PEN), cytosolic manganese superoxide dismutase (cMnSOD), and lysozyme. Transcripts of all nine genes were detected in all tissues with differential expression levels when examined by RT-PCR and qPCR. BGBP-HDL, LGBP and haemocyanin were mainly expressed in the hepatopancreas and their expressions levels were about 1/10-1/3 those of beta-actin. Their expressions in other tissues were relatively limited. ProPO, TGase, crustins, PEN-3, and lysozyme showed the highest levels of expression in haemocytes and the lowest in hepatopancreas. Their expression levels in the haemocytes were 3 (PEN-3) to 10(-2) (proPO) times those of beta-actin. In contrast to the other genes, cMnSOD showed higher expression levels in haemolymph related organ, stomach and muscle; and lower expression levels in haemocyte, migut, neural ganglion and hepatopancreas. When examined by in situ hybridisation, hepatopancreatic F cells were found to be the major cell type that produced transcripts of BGBP-HDL, LGBP and haemocyanin. On the other hand, circulatory haemocytes and haemocytes infiltrated in various tissues contributed to the expressions of proPO, TGase, crustins, PEN-3 and lysozyme. Both hepatopancreatic F cell and haemocyte generated cMnSOD transcripts. Using in situ hybridisation, the present study is the first to show the tissue distributions of BGBP-HDL, LGBP, haemocyanin, TGase, crustins and cMnSOD in healthy white shrimp. The present results provide a baseline data of physiological expressions for the genes that are important in immune activation and modulation in Pacific white shrimp and a guideline of tissue or organ sampling for effective gene expression analyses for future immunological studies.


Four-Year Follow-up of [18 F]Fluorodeoxyglucose Positron Emission Tomography-Based Parkinson's Disease-Related Pattern Expression in 20 Patients with Isolated Rapid Eye Movement Sleep Behavior Disorder Shows Prodromal Progression.
Isolated rapid eye movement sleep behavior disorder is known to be prodromal for alpha-synucleinopathies, such as Parkinson's disease (PD) and dementia with Lewy bodies. The [18 F]fluorodeoxyglucose-positron emission tomography (PET)-based PD-related brain pattern can be used to monitor disease progression. We longitudinally investigated PD-related brain pattern expression changes in 20 subjects with isolated rapid eye movement sleep behavior disorder to investigate whether this may be a suitable technique to study prodromal PD progression in these patients and to identify potential phenoconverters. Subjects underwent two [18 F]fluorodeoxyglucose-PET brain scans ~3.7 years apart, along with baseline and repeated motor, cognitive, and olfactory testing within roughly the same time frame. At baseline, 8 of 20 (40%) subjects significantly expressed the PD-related brain pattern (with z scores above the receiver operating characteristic-determined threshold). At follow-up, six additional subjects exhibited significant PD-related brain pattern expression (70% in total). PD-related brain pattern expression increased in all subjects (P = 0.00008). Four subjects (20%), all with significant baseline PD-related brain pattern expression, phenoconverted to clinical PD. Suprathreshold PD-related brain pattern expression and greater score rate of change may signify greater shorter-term risk for phenoconversion. Our results support the use of serial PD-related brain pattern expression measurements as a prodromal PD progression biomarker in patients with isolated rapid eye movement sleep behavior disorder. © 2020 The Authors. Movement Disorders published by Wiley Periodicals LLC on behalf of International Parkinson and Movement Disorder Society.


Survey data on entrepreneurs׳ subjective plan and perceptions of the likelihood of success.
Entrepreneurship is an important economic process in both developed and developing worlds. Nonetheless, many of its concepts appear to be difficult to 'operationalize' due to lack of empirical data; and this is particularly true with emerging economy. The data set described in this paper is available in Mendeley Data׳s "Vietnamese entrepreneurs' decisiveness and perceptions of the likelihood of success/continuity, Vuong (2015) [1]" http://dx.doi.org/10.17632/kbrtrf6hh4.2; and can enable the modeling after useful discrete data models such as BCL.


Acyanotic and Cyanotic Congenital Heart Diseases.
There has been tremendous progress in treatment of heart disease in children. Device therapy is increasingly being used in acyanotic congenital heart disease, while surgical results have improved significantly to give smile to many cyanotic heart disease children and their parents. This article makes an attempt to increase awareness of general pediatricians about common congenital heart diseases.


Comparison between laser- and diathermy-assisted posterior cordotomy for bilateral vocal cord abductor paralysis.
A reliable procedure is needed to solve the problem of difficult airway in patients with bilateral vocal cord paralysis without adversely affecting patient "voice quality." To compare the results of laser-assisted posterior cordotomy with diathermy-assisted posterior cordotomy for bilateral vocal cord paralysis in regard to dyspnea severity, voice quality, and aspiration. Prospective randomized clinical trial at a university medical center. Zagazig University Hospitals, Zagazig, Egypt. Twenty patients randomly categorized into 2 groups; group A was treated with laser-assisted posterior cordotomy and group B was treated with diathermy-assisted posterior cordotomy. Laser-assisted posterior cordotomy for group A and diathermy-assisted posterior cordotomy for group B. Dyspnea severity, voice quality, and aspiration. A significant difference (P < .05) was found between group A and group B at all postoperative comparisons in dyspnea, whereas no significant difference (P ≥ .05) was detected at all postoperative comparisons in voice quality. Laser-assisted posterior cordotomy can be considered as a reliable and superior procedure compared with diathermy-assisted posterior cordotomy in the treatment of bilateral vocal cord abductor paralysis. clinicaltrials.gov Identifier: ISRCTN08093874.


Effect of hyperosmotic conditions on the expression of the betaine-GABA-transporter (BGT-1) in cultured mouse astrocytes.
The adaptation of cells to hyperosmotic conditions involves accumulation of organic osmolytes to achieve osmotic equilibrium and maintenance of cell volume. The Na+ and Cl(-)-coupled betaine/GABA transporter, designated BGT-1, is responsible for the cellular accumulation of betaine and has been proposed to play a role in osmoregulation in the brain. BGT-1 is also called GAT2 (GABA transporter 2) when referring to the mouse transporter homologue. Using Western Blotting the expression of the mouse GAT2 protein was investigated in astrocyte primary cultures exposed to a growth medium made hyperosmotic (353+/-2.5 mosmol/kg) by adding sodium chloride. A polyclonal anti-BGT-1 antibody revealed the presence of two characteristic bands at 69 and 138 kDa. When astrocytes were grown for 24 h under hyperosmotic conditions GAT2 protein was up-regulated 2-4-fold compared to the level of the isotonic control. Furthermore, the expected dimer of GAT2 was also up-regulated after 24 h under the hyperosmotic conditions. The [3H]GABA uptake was examined in the hyperosmotic treated astrocytes, and characterized using different selective GABA transport inhibitors. The up-regulation of GAT2 protein was not affecting total GABA uptake but the hyperosmotic condition did change total GABA uptake possibly involving GAT1. Immunocytochemical studies revealed cell membrane localization of GAT2 throughout astroglial processes. Taken together, these results indicate that astroglial GAT2 expression and function may be regulated by hyperosmolarity in cultured mouse astrocytes, suggesting a role of GAT2 in osmoregulation in neural cells.


Biogenic stabilization and heavy metal immobilization during vermicomposting of vegetable waste with biochar amendment.
Vermicomposting is a traditional technology that produces the best quality of compost, but factors such as maturity, presence of heavy metals, etc. need to be tackled prior to agrarian application. The present study investigates the influence of varying biochar dose (2.5, 5, and 10% on a weight basis) on the maturity of compost and heavy metals during vermicomposting of vegetable waste using epigeic earthworm. Biochar amendment notably enhanced the electrical conductivity (up to 2.7 mS/cm), nitrogen content (up to 3.1%), NO3-N (up to 630 mg/kg) and nutritional value. The heavy metals, oxygen uptake rate (below 0.96 mg/g VS/day) and CO2 evolution rate (below 1 mg/g VS/day) were attenuated along with degradation of complex organic crystals as observed in powder X-Ray Diffraction (PXRD) spectra. Furthermore, biochar aid in reducing pathogens (below 1.1 × 103 MPN/g dry weight) as inferred from the Most Probable Number (MPN) results as well as degrading the complex organics into simpler compounds as revealed from the Fourier-transform infrared spectroscopy (FTIR) spectra. The present study inferred that the vegetable waste was biologically stabilized through biochar amendment during vermicomposting process with improved nutritional and physico-chemical properties.


Development and technical validation of a smartphone-based pediatric cough detection algorithm.
Coughing is a common symptom in pediatric lung disease and cough frequency has been shown to be correlated to disease activity in several conditions. Automated cough detection could provide a noninvasive digital biomarker for pediatric clinical trials or care. The aim of this study was to develop a smartphone-based algorithm that objectively and automatically counts cough sounds of children. The training set was composed of 3228 pediatric cough sounds and 480,780 noncough sounds from various publicly available sources and continuous sound recordings of 7 patients admitted due to respiratory disease. A Gradient Boost Classifier was fitted on the training data, which was subsequently validated on recordings from 14 additional patients aged 0-14 admitted to the pediatric ward due to respiratory disease. The robustness of the algorithm was investigated by repeatedly classifying a recording with the smartphone-based algorithm during various conditions. The final algorithm obtained an accuracy of 99.7%, sensitivity of 47.6%, specificity of 99.96%, positive predictive value of 82.2% and negative predictive value 99.8% in the validation dataset. The correlation coefficient between manual- and automated cough counts in the validation dataset was 0.97 (p < .001). The intra- and interdevice reliability of the algorithm was adequate, and the algorithm performed best at an unobstructed distance of 0.5-1 m from the audio source. This novel smartphone-based pediatric cough detection application can be used for longitudinal follow-up in clinical care or as digital endpoint in clinical trials.


A new method with fiberoptic transducers used for simultaneous recording of intravesical and urethral pressure during physiological filling and voiding phases.
A new technique using fiberoptic transducers for simultaneous registration of intravesical and urethral pressures during the physiological filling and voiding phases is described. The method, which is simple, exact and stable, was used on several patients with different urological problems. The study shows that the measurement technique and the fiberoptic transducer system are suited for clinical use.


Treatment of childhood acute immune thrombocytopenic purpura with anti-D immune globulin or pooled immune globulin.
To evaluate the effectiveness of initial treatment of children with acute immune thrombocytopenic purpura (ITP) with anti-D immune globulin (anti-D) or pooled IgG immune globulin (IVIg). The medical charts of 33 children diagnosed with acute ITP from May 1995 to October 1997 were reviewed. Patient data were eligible for analysis if, for the new diagnosis of acute ITP, the patient had received either anti-D at 45 to 50 microg/kg (WinRho SD, NABI) or IVIg at 0.8 to 1 g/kg (Gammagard SD, Baxter-Highland). The platelet response time for each treatment group was compared by the Mann-Whitney U test. Time to achieve a platelet count >/=20 x 10(9 )/L (20,000/mm3 ) was 1.54 +/- 0.51 days in the IVIg group (n = 13) and 1.26 +/- 0.82 days in the anti-D group (n = 14) (P =.34). Time to achieve a platelet count >/=40 x 10(9 )/L (40,000/mm3 ) was 1.77 +/- 0.74 and 1.49 +/- 1.01 days for the IVIg and anti-D groups, respectively (P =.32). Children given IVIg were hospitalized for 2.1 +/- 0.87 days, whereas those given anti-D were hospitalized for 1.94 +/- 1.08 days. A net decrease in hemoglobin concentration was observed after receipt of IVIg (9.1 +/- 7.3 g/L [0.91 +/- 0.73 g/dL]) and after anti-D therapy (4.5 +/- 10.3 g/L [0.45 +/- 1.03 g/dL], P =.23). No patient required intervention for hemolysis. In this retrospective analysis anti-D was as effective as IVIg for the treatment of acute ITP in children. However, randomized, controlled trials are needed to establish the role of anti-D in the treatment of acute ITP in children.


Kinetic studies on the reactivation of D-beta-hydroxybutyrate dehydrogenase with mixtures of short-chain lecithins.
D-beta-hydroxybutyrate dehydrogenase, purified as soluble, lipid-free apoenzyme (inactive) from rat liver mitochondria can be reactivated by the short-chain dihexanoyl, diheptanoyl, and dioctanoyl lecithins at the monomeric state, upon formation of a reversible enzyme-lecithin complex. Previous studies with these lecithins suggested that reactivation of the apoenzyme requires the simultaneous occupation of two identical, noninteracting lecithin binding sites via a rapid equilibrium random mechanism. The short-chain lecithins exhibited similar reactivating capacities, differing only in their affinities towards the enzyme. In order to further test that model, the reactivation of the apoenzyme was studied when two or three short-chain lecithins were simultaneously present in the reaction medium. The initial velocities were measured either as a function of the concentration of one lecithin while the other(s) were kept constant, or as a function of the total phospholipid concentration with mixtures of different lecithins at a constant molar ratio. The pertinent equations were derived on the principles of multiple equilibria with identical, noninteracting sites able to be occupied by any of the different lecithins present in the reaction medium, with the doubly occupied enzyme as the only active species. In agreement with the above-proposed model, the results obtained indicates that the molar fraction of the doubly occupied (active) enzyme species can be calculated from equilibrium considerations and that the maximal attainable with the different short-chain lecithins are similar.


Felodipine-induced inhibition of polymorphonuclear leukocyte functions.
Felodipine inhibits fMet-Leu-Phe or ionophore A23187-induced exocytosis in rabbit peritoneal polymorphonuclear leukocytes (PMNs), in the concentration range 1-50 microM. Activation of the metabolic burst, and migration of PMNs towards fMet-Leu-Phe are equally inhibited by felodipine in the same concentration range. The effect is not due to blocking of calcium channels in the plasma membrane, because the degree of inhibition remains the same when Ca2+ is omitted from the medium. Felodipine interferes with ionophore A23187-induced association of 45Ca with the PMN but this interference occurs at lower concentrations than the inhibition of exocytosis. Hypotonic hemolysis of erythrocytes is inhibited by felodipine; maximal protection against hemolysis occurs at a concentration of 50 microM felodipine. It is suggested that at least a part of the inhibiting effect on PMN functions might be due to an anesthetic-like membrane effect of felodipine.


Characterization of the temperature- and pressure-induced inverse and reentrant transition of the minimum elastin-like polypeptide GVG(VPGVG) by DSC, PPC, CD, and FT-IR spectroscopy.
We investigated the temperature- and pressure-dependent structure and phase behavior of a solvated oligopeptide, GVG(VPGVG), which serves as a minimalistic elastin-like model system, over a large region of the thermodynamic phase field, ranging from 2 to 120 degrees C and from ambient pressure up to approximately 10 kbar, applying various spectroscopic (CD, FT-IR) and thermodynamic (DSC, PPC) measurements. We find that this octapeptide behaves as a two-state system which undergoes the well-known inverse-temperature folding transition occurring at T approximately 36 degrees C, and, in addition, a slow trend reversal at higher temperatures, finally leading to a reentrant unfolding close to the boiling point of water. Furthermore, the pressure-dependence of the folding/unfolding transition was studied to yield a more complete picture of the p, T-stability diagram of the system. A molecular-level picture of these processes, in particular on the role of water for the folding and unfolding events of the peptide, presented with the help of molecular-dynamics simulations, is presented in a companion article in this issue.


Determinants of influenza and COVID-19 vaccine intent or uptake in Lebanon: a scoping review of the literature.
Vaccination is essential to protect from influenza and recently from COVID-19, yet uptake in Lebanon is suboptimal. Several factors determine uptake including knowledge, attitude and policies. We conducted a scoping review of the literature to explore the determinants of influenza and COVID-19 vaccine intent or uptake in Lebanon. Following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines, thirty one peer reviewed studies indexed in six databases Pub Med, EMBASE, Scopus, CINAHL, Medline, and the Cochrane Library were screened. Two students, a senior Librarian and an Associate Professor in nursing searched for eligible studies. The library search strategy followed a combination of three broad concepts (viral influenzas; vaccines; Lebanon). The search timeframe was up till December 31, 2022. Determinants of influenza and COVID-19 vaccine intent or uptake were categorized following the constructs of the Theory of Planned Behavior. Nine studies investigated influenza vaccine intent or uptake among the public community, parents and healthcare workers. Twenty one studies investigated COVID-19 vaccine intent or uptake among the public community, older refugees, university students, patients with cancer, dentists, and social media users. One study investigated both types of vaccines. A conceptual model of the determinants of vaccine intent and uptake within the Lebanese context was developed. Various determinants included environmental factors, norms, knowledge, perceptions, attitudes, past experiences, behavioral control and hesitancy. Research on vaccine intent and uptake in Lebanon is still in its infancy, while that of COVID-19 is on the rise. Multifaceted reasons behind the low vaccination rates were observed yet few attempts were made to target vulnerable groups. Further research studies are needed to target vulnerable groups.


SNC1, a yeast homolog of the synaptic vesicle-associated membrane protein/synaptobrevin gene family: genetic interactions with the RAS and CAP genes.
SNC1, a gene from the yeast Saccharomyces cerevisiae, encodes a homolog of vertebrate synaptic vesicle-associated membrane proteins (VAMPs) or synaptobrevins. SNC1 was isolated by its ability to suppress the loss of CAP function in S. cerevisiae strains possessing an activated allele of RAS2. CAP is a component of the RAS-responsive S. cerevisiae adenylyl cyclase complex. The N-terminal domain of CAP is required for full cellular responsiveness to activated RAS proteins. The C-terminal domain of CAP is required for normal cellular morphology and responsiveness to nutrient extremes. Multicopy plasmids expressing SNC1 suppress only the loss of the C-terminal functions of CAP and only in the presence of activated RAS2.


Systems biology of autophagy in leishmanial infection and its diverse role in precision medicine.
Autophagy is a contentious issue in leishmaniasis and is emerging as a promising therapeutic regimen. Published research on the impact of autophagic regulation on Leishmania survival is inconclusive, despite numerous pieces of evidence that Leishmania spp. triggers autophagy in a variety of cell types. The mechanistic approach is poorly understood in the Leishmania parasite as autophagy is significant in both Leishmania and the host. Herein, this review discusses the autophagy proteins that are being investigated as potential therapeutic targets, the connection between autophagy and lipid metabolism, and microRNAs that regulate autophagy and lipid metabolism. It also highlights the use of systems biology to develop novel autophagy-dependent therapeutics for leishmaniasis by utilizing artificial intelligence (AI), machine learning (ML), mathematical modeling, network analysis, and other computational methods. Additionally, we have shown many databases for autophagy and metabolism in Leishmania parasites that suggest potential therapeutic targets for intricate signaling in the autophagy system. In a nutshell, the detailed understanding of the dynamics of autophagy in conjunction with lipids and miRNAs unfolds larger dimensions for future research.


Haemodynamic effects of molsidomin.
Hemodynamic changes for 30 min after intravenous administration of molsidomin 4 mg were studied in 10 patients. Pulmonary artery pressure, pulmonary wedge pressure, right atrial pressure and aortic pressure decreased significantly after 15 min, and after 30 min they were still below the control level. Heart rate increased significantly (by 15%) 15 min after injection, and after 30 min it was still 8% above the control value. Cardiac index decreased significantly from 3.37 +/- 0.12 to 2.95 +/- 0.15 1/min/m2. Peripheral and pulmonary resistance showed no significant change. Both stroke work index and minute work decreased significantly 15 min after administration of molsidomin and after 30 min both were still below the control level. The results suggest that molsidomin decreases preload without influencing afterload. By decreasing preload, molsidomin improves the oxygen supply/demand ratio of the myocardium.


Secure medical image encryption with Walsh-Hadamard transform and lightweight cryptography algorithm.
It is important to ensure the privacy and security of the medical images that are produced with electronic health records. Security is ensured by encrypting and transmitting the electronic health records, and privacy is provided according to the integrity of the data and the decryption of data with the user role. Both the security and privacy of medical images are provided with the innovative use of lightweight cryptology (LWC) and Walsh-Hadamard transform (WHT) in this study. Unlike the light cryptology algorithm used in encryption, the hex key in the algorithm is obtained in two parts. The first part is used as the public key and the second part as the user-specific private key. This eliminated the disadvantage of the symmetric encryption algorithm. After the encryption was performed with a two-part hex key, the Walsh-Hadamard transform was applied to the encrypted image. In the Walsh-Hadamard transform, the Hadamard matrix was rotated with certain angles according to the user role. This allowed the encoded medical image to be obtained as a vector. The proposed method was verified with the results of the number of pixel change rates and unified average changing intensity measurement parameters and histogram analysis. The results showed that the method is more successful than the lightweight cryptology method and the proposed methods in the literature to solve security and privacy of the data in medical applications with user roles.


Multiplex qPCR assay for ultra sensitive detection of JCV DNA with simultaneous identification of genotypes that discriminates non-virulent from virulent variants.
JC virus (JCV) is the etiologic agent for progressive multifocal leukoencephalopathy (PML), a demyelinating disease occurring in the brain of patients with underlying immune compromised states. All viable JCV genomes contain a conserved region in the T protein coding nucleotide sequence that when detected by PCR in CSF is a confirmatory diagnostic marker for PML along with clinical and neuroradiological evidence. The non-coding regulatory region (NCRR) is hypervariable, as evidenced by nucleotide sequence of the non-virulent variant, which is predominantly excreted in urine, versus that of virulent variants found in brain and CSF of PML patients. All variants can be found in blood. A single assay that quantifies and identifies JCV DNA in clinical samples and discriminates between variants has significant value to physicians and patients at risk for PML. Separate primer pairs were tested together to quantitatively detect conserved viral DNA nucleotide sequence in patient samples, while simultaneously detecting the NCRR specific for the non-virulent variant. In testing using control plasmids and patients' CSF, blood, and urine, PML patients predictably demonstrated the non-virulent, archetype NCRR in urine, but virulent NCRR variants in CSF and blood. The JCV qPCR multiplex assay targets two regions in JCV genomes to simultaneously identify and measure viral DNA, as well as distinguish between variants associated with PML and those that are not. The multiplex results could signal risk for PML if patients are viremic with JCV variants closely associated with PML pathogenesis.


Plasma metabolomic profiling of repeated restraint stress in rats.
Exposure to repeated stress is associated with the occurrence and development of many prevalent diseases. Here, we evaluated the impact of a repeated restraint stress (RRS) paradigm on male Wistar rats and compared global plasma metabolites in stressed and unstressed rats using an ultra-high performance liquid chromatography-quadrupole time-of-flight mass spectrometry (UHPLC/Q-TOF-MS) approach and multivariate statistical analysis. The data showed that ten days of RRS induced weight loss and anxiety-like behavior, possibly through elevating plasma corticosterone levels in rats. Different metabolic characteristics were found between the two groups. Seventeen differential metabolites were identified and considered as potential biomarkers in the stress process, including histidine metabolism, neurotransmitter synthesis, and bile acid metabolism. Furthermore, l-carnosine, an identified metabolite, was examined in the same model and found to attenuate weight loss and anxiety-like behavior in RRS rats. Hence, these findings describe the metabolomic profiling of repeated stress and provide new insights into the pathophysiological mechanism of stress-induced deflections and potential pharmacological interventions.


Determination of ochratoxin A at the ppt level in human blood, serum, milk and some foodstuffs by high-performance liquid chromatography with enhanced fluorescence detection and immunoaffinity column cleanup: methodology and Swiss data.
An improved specific analytical method for ochratoxin A (OA) is presented, combining HPLC separation with enhanced fluorescence detection by post-column addition of ammonia. Commercial immunoaffinity columns (Biocode) were for the first time applied to the cleanup of extracts of body fluids; they could be used up to 20 times for blood serum. The extraction efficiency of OA from human serum and milk as well as its derivatization to esters were studied and improved. The quantitation limit for OA was improved and estimated at 5-10 pg/g for human milk and serum. The mean recovery of OA from serum and milk was estimated at 85%. The overall coefficient of variation for OA determinations in serum, milk and selected foodstuffs was estimated at 10% (concentration range 0.01-5 ng/g). The method was applied to sera of 368 blood donors, 10 pairs of maternal and fetal sera, as well as to 40 human milk samples and selected foodstuffs; the results are discussed.


Scientism and Pseudoscience: A Philosophical Commentary.
The term "scientism" is used in a variety of ways with both negative and positive connotations. I suggest that some of these uses are inappropriate, as they aim simply at dismissing without argument an approach that a particular author does not like. However, there are legitimate negative uses of the term, which I explore by way of an analogy with the term "pseudoscience." I discuss these issues by way of a recent specific example provided by a controversy in the field of bioethics concerning the value, or lack thereof, of homeopathy. I then frame the debate about scientism within the broader context of C.P. Snow's famous essay on the "two cultures."


Cerebral infarction in a young man using high-dose anabolic steroids.
Anabolic androgenic steroid (AAS) abuse has increased among athletes in recent years. However, AAS abuse can increase hypercoagulopathy and cause cerebrovascular disease. We report a case of a 27-year-old man who had right hemiparalysis, hemianopia, dysarthria, and double vision in the middle of muscle training. He suspected acute disseminated encephalomyelitis at first, because of a preceding respiratory infection. However, extensive work-up was performed, including brain magnetic resonance imaging, transcranial Doppler and transesophageal echocardiography, confirming the final diagnosis of cardioembolic stroke. Physicians should be aware that cerebrovascular disease may be a side effect of AAS, even in younger populations.


Problem-solving instruction during the clinical clerkship: description and preliminary evaluation of a programme.
A clerkship programme in general surgery is described whereby students acquire factual knowledge about disease entities through independent learning rather than through formal teaching. The objectives of the programme are to promote problem-solving skills. It consists of clinical work (patient work-up, clinical rounds, participation in the activities in the operating room, out-patient clinic and emergency room) and of small-group tutorial sessions. The described programme has met with approval and satisfaction from both students and teachers. There were no significant differences in the average grades on the written and oral final examinations between the graduates of this clerkship programme and those of a traditional one. Independent learning and problem-solving instruction, which have been adopted by some medical schools in the preclinical phase of the curriculum, can be effectively implemented during the clinical phase as well.


Bone mechanotransduction may require augmentation in order to strengthen the senescent skeleton.
Physical exercise is thought to hold promise as a non-invasive countermeasure against skeletal fragility arising from post-menopausal and age-related osteoporosis. Importantly, mechanical loading and exercise are capable of increasing bone size via periosteal expansion, which by far, is the most effective means of strengthening the structure of a given bone. The focus of this review was to therefore explore whether exercise has the potential to increase periosteal modeling and bone size in the senescent skeleton. A survey of exercise trials in humans suggests that exercise interventions that enhance periosteal modeling in the young skeleton fail to do the same in the elderly skeleton. Underlying this ineffectiveness, in vitro studies indicate that aging lowers basal levels of cell function and degrades bone mechanotransduction at a variety of levels from altered second messenger signaling to gene expression driving proliferation and/or differentiation. Given these age-related alterations, the ultimate efficacy of an exercise intervention may depend upon concurrent supplementation that directly address deficits in signaling and/or cell function. In this context, in vivo animal models of mechanical loading that simulate the muted periosteal adaptation in the elderly hold potential to examine the efficacy of countermeasures. Preliminary in vivo experiments suggest that pharmacologically counteracting age-related deficits in cellular function can restore exercise induced periosteal modeling in the senescent skeleton to levels observed in young animals. If the safety and efficacy of this strategy were to be confirmed for human use, it would enable the utilization of exercise as a viable countermeasure against skeletal fragility at senescence.


A case report of xanthogranulomatous osteomyelitis of the distal ulna mimicking a malignant neoplasm.
Male, 59 FINAL DIAGNOSIS: Xanthogranulomatous osteomyelitis Symptoms: Painful swelling in the wrist Medication: Drug history of antifulgal agents Clinical Procedure: Excisional biopsy Specialty: Radiology. Rare disease. Xanthogranulomatous inflammation is a chronic inflammatory disease in which bone involvement is extremely rare. Bone involvement of xanthogranulomatous inflammation, termed xanthogranulomatous osteomyelitis (XO), often presents as a mass-like lesion extending to adjacent structures, which can mimic infiltrative carcinoma. We present a case of XO in the ulna, which mimicked a neoplasm. A 59-year-old man presented with a 2-month history of painful swelling in the right wrist. Plain radiography and CT showed an osteolytic lesion at the distal ulna. MRI revealed a soft-tissue mass with lobulated margins and contrast enhancement. Intense uptake in bone scan and PET suggested malignancy. An excisional biopsy from the representative area resulted in a pathology diagnosis of XO. Gross and radiologic manifestations of XO can mimic neoplasm. XO generally has benign prognosis, contrary to malignant bone tumor. Therefore, biopsy and histopathological confirmation are necessary for proper management.


Three-Dimensional Imaging of Urethral Stricture Disease and Urethral Pathology for Operative Planning.
Diagnosing urethral pathology can prove difficult, as clinically, the presentation is often nonspecific and may be suggestive of multiple etiologies. Therefore, detailed and accurate urethral imaging in both males and females is critical. Since the early 1900s, conventional imaging studies including RUG and VCUG, with adjunct cystourethroscopy, have remained the gold standard diagnostic techniques to evaluate urethral pathology. However, limitations of conventional imaging have generated interest in finding alternative imaging modalities with comparable, if not superior, diagnostic accuracy, the goal being a more complete assessment of urethral pathology and anatomy that would allow for appropriate surgical planning. Imaging modalities with three-dimensional (3D) capabilities may provide more comprehensive information regarding urethral diseases through a more detailed illustration of periurethral soft tissue structures. Whether or not these imaging modalities will replace conventional studies is unclear, though there is an increasing body of literature that support their use.


Photodynamic treatment of oral squamous cell carcinoma in hamster cheek pouch model using chlorin p6-histamine conjugate.
Over-expression of histamine receptors has been reported in several types of malignancies. Earlier we have successfully demonstrated use of chlorin p6-histamine conjugate (Cp6-his) for improving cellular uptake and photo toxicity of Cp6 in oral cancer cell lines. In the present study, after having confirmed that histamine receptors are over-expressed in tumors of hamster cheek pouch, we investigated the efficacy of Cp6-his for photodynamic treatment (PDT) of tumors in this animal model. Cp6-his (3mg/kg body weight) was injected intraperitoneally and its accumulation in tumor, surrounding tissue, normal mucosa and abdominal skin was monitored non-invasively by fluorescence spectroscopy. For PDT, tumors at 4h after Cp6-his administration were exposed to red light (660±25nm, 100J/cm(2)). Tumor damage and regression were assessed by histology and tumor volume measurements, respectively. Expression of histamine H2 receptors in tumor and normal mucosa was assessed by immuno-staining. The accumulation of Cp6-his was higher in tumors as compared to normal mucosa at 4h after its administration. For Cp6 similar preferential accumulation was observed except that in normal mucosa the accumulation of Cp6 was more as compared to Cp6-his. The clearance of Cp6-his from skin was rapid showing ∼80% decrease within 48h from its peak level at 4h after drug injection. PDT led to extensive cellular damage and tumors of size up to ∼1000mm(3) regressed completely one week after PDT. Higher tumor selectivity of Cp6-his and complete regression of bigger tumors after PDT suggest that conjugating Cp6 to histamine is a promising approach to improve PDT efficacy.


A novel mutation at a probable heme-binding ligand in neutrophil cytochrome b558 in atypical X-linked chronic granulomatous disease.
A membrane-bound cytochrome b558, a heterodimer consisting of gp91-phox and p22-phox, is a critical component of the superoxide (O2-)-generating reduced nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in phagocytes. Chronic granulomatous disease (CGD) is characterized by recurrent bacterial infection caused by a defect of the oxidase. Both subunits are absent from phagocytes in typical X-linked recessive CGD patients who are primarily defective in gp91-phox. We report here an atypical case of X-linked CGD in which neutrophils showed a complete absence of O2--forming NADPH oxidase activity, but a small amount (about 10% of control) of both subunits was detected by immunoblot analysis. Spectrophotometric studies of the neutrophils with a recently developed sensitive method gave no evidence for the heme spectrum in the cytochrome b558, of this CGD. Reverse transcription/polymerase chain reaction and sequence analysis revealed a C to T transition replacing histidine at amino acid position 101 (His101) by tyrosine in gp91-phox. These results provide evidence that His101 of gp91-phox is the one of the heme-binding ligands of cytochrome b558.


Time related changes of T1, T2, and T2(*)(2) of human blood in vitro.
In view of a potential future use for dating hemorrhage in forensic medicine the correlation of MR relaxation parameters with time was evaluated in blood samples. A systematic relationship could be valuable for using MRI for estimating the age of hemorrhage and soft tissue hematomas particularly in clinical forensic medicine. Relaxation times T1, T2, and T2(*) of venous blood samples from 6 volunteers were measured using 3T MRI regularly up to 30 days. The time progression of the relaxation parameters was systematically analyzed and examined for possible interrelations. T2 initially decreased to a minimum, and then increased again (range 24-97ms), while T1 started with a plateau phase followed by an almost linear decrease (range 333-2153ms). T2(*) remained relatively constant during the entire investigation period. The higher the initial T2 was, the lower was its minimum, and the greater was the decrease of the associated T1. The inter- and intra-individual variability was relatively large, one reason being very likely the metabolic differences in the blood samples. The observed characteristic changes in blood samples over time measured by quantitative MR techniques add objective information in view of an estimation of the age of hemorrhage. However, in vivo studies will be needed to verify the data with respect to influencing metabolic factors.


CHADS(2) score is predictive of left atrial thrombus on precardioversion transesophageal echocardiography in atrial fibrillation.
The goals of this study were to determine: 1) if the CHADS(2) score correlates with left atrial (LA) or left atrial appendage (LAA) thrombus on pre-cardioversion transesophageal echocardiography (TEE) in nonvalvular atrial fibrillation (NVAF); and 2) what, if any, components of the CHADS(2) score are most important in predicting LA/LAA thrombus. It is unknown if CHADS(2) score, a marker of thromboembolic risk in NVAF, accurately predicts LA/LAA thrombus on pre-cardioversion TEE. We retrospectively studied patients undergoing precardioversion TEE for NVAF at a tertiary hospital. TEE reports were reviewed for presence of LA/LAA thrombus. Using medical records and an ICD-9 coding database, a CHADS(2) score was derived, and the association between CHADS(2) and thrombus was evaluated with Mantel-Haenszel Chi-Square. The relation between the singular components of CHADS(2) and thrombus were analyzed using Pearson's Chi-Square. In 643 consecutive patients undergoing pre-cardioversion TEE, LA/LAA thrombus was identified in 46 (7.2 %). A strong association was present between CHADS(2)score and LA/LAA thrombus (p = 0.0005). No thrombi were identified in patients with CHADS(2) = 0. Among 46 patients with thrombus, all (100%) had CHF. Of the singular components, CHF was the only factor independently associated with thrombus (p < 0.0001). In non-valvular atrial fibrillation, CHADS(2) is strongly associated with LA thrombus on TEE. Our findings suggest pre-cardioversion TEE may be unnecessary if the CHADS(2) score = 0. Of the components of the CHADS(2) score, CHF was the only independently associated risk factor which correlated with LA/LAA thrombus.


Risk scoring system for prediction of obstetric anal sphincter injury.
The objective was to begin the process of developing an antenatal risk scoring system, as a first step towards examining whether elective Caesarean section for women at high risk of injury could be an effective and acceptable intervention. Retrospective study. Tertiary maternity unit in the UK. One hundred and twenty-three women who sustained an obstetric anal sphincter injury (OASI) and 123 controls without OASI. Case notes of women with a third or fourth degree tear between 1997 and 1999 were examined for risk factors. Controls matched for age and week of delivery were identified from the maternity record database and case records reviewed for the presence of risk factors. Unweighted and weighted risk scores were produced using odds ratios, and compared between cases and controls. Receiver operating characteristics (ROC) curve analysis of the risk scores was performed to discriminate between cases and controls and to calculate the sensitivity and specificity of each scoring system. Odds ratio (OR) and 95% confidence interval (CI) for each risk factor. Sensitivity and specificity from ROC curves for weighted and unweighted risk score. Among the cases there were more nulliparous women (OR 1.77; CI 1.05-2.99) and a trend towards more women with an episiotomy (OR 1.57; CI 0.99-2.47). Among women with sphincter injury, trends towards more epidurals (OR 1.64; CI 0.97-2.75), and more babies weighing more than 4000 g among (OR 1.45; CI 0.85-2.49) were noted. The median unweighted risk score was 2 for cases and 2 for controls (P= 0.05), while the weighted risk score was 2.1 and 1.37 (P= 0.03), respectively. The ROC curves approximated to a straight line demonstrating very poor discrimination between cases and controls. The predictive test performed poorly, suggesting that the risk factors identified do not exert a large enough effect in a cohort of this size.


SMC proteins and chromosome structure.
The structure of chromosomes is largely determined by chromosome-associated proteins. Members of the SMC (structural maintenance of chromosomes) family play an important role in both prokaryotic and eukaryotic chromosome structure and dynamics. SMC proteins are involved in chromosome condensation, sister-chromatid cohesion, sex-chromosome dosage compensation, genetic recombination and DNA repair. There have been major advances recently in understanding the function of SMC proteins--including the identification of biochemical activities of SMC-containing protein complexes and the realization that individual SMC proteins might link seemingly unrelated aspects of chromosomal metabolism.


Novel cells integrated biosensor based on superoxide dismutase on electrospun fiber scaffolds for the electrochemical screening of cellular stress.
A novel electrochemical biosensor was developed to monitor fibroblast cells stress levels for the first time in situ under external stimuli based on the recognition of superoxide anion released upon cell damage. The biosensor comprised metallized polycaprolactone electrospun fibers covered with zinc oxide for improved cell adhesion and signal transduction, whilst stable bioconjugates of mercaptobenzoic acid-functionalized gold nanoparticles/superoxide dismutase were employed as recognition bioelements. Biosensors were first tested and optimized for in situ generated superoxide detection by fixed potential amperometry at +0.3 V, with minimal interferences from electroactive species in cell culture media. L929 fibroblast cells were then implanted on the optimized biosensor surface and the biosensor morphologically characterized by scanning electron microscopy (SEM) and fluorescence microscopy, which illustrated the network-type pattern of fibroblasts adjacent to the fiber scaffold. Fibroblast stress was induced by zymosan and monitored at the cells integrated biosensor using fixed potential amperometry (CA) with a sensitivity of 26 nA cm-2 μg mL-1 zymosan and electrochemical impedance spectroscopy (EIS), with similar sensitivity of the biosensor considering the Rs and Z' parameters of around 0.13 Ω cm2 μg-1 mL and high correlation factors R2 of 0.9994. The obtained results underline the applicability of the here developed biosensor for the electrochemical screening of the fibroblast cells stress. The concept in using low-cost biocompatible polymeric fibers as versatile scaffolds for both enzyme immobilization and cell adhesion, opens a new path in developing biosensors for the in-situ investigation of a variety of cellular events.


CAG repeat length in androgen receptor gene is not associated with amyotrophic lateral sclerosis.
Epidemiological and clinical studies show higher prevalence of amyotrophic lateral sclerosis (ALS) in males than in females and more severe lesions in androgen receptor (AR)-expressing tissues. The AR gene contains a polymorphic CAG trinucleotide repeat, whose expansion over a certain threshold is toxic to motor neurons, causing spinal and bulbar muscular atrophy (SBMA). We tested the hypothesis that the AR CAG repeat linked to SBMA is a risk factor for ALS. We analyzed AR CAG expansions in 336 patients with ALS and 100 controls. We found a negative association of AR CAG expansions with ALS susceptibility, clinical presentation, and survival. Our findings do not support a role of the AR CAG repeat length in ALS.


The role of imaging for the management of newly diagnosed glioblastoma in adults: a systematic review and evidence-based clinical practice guideline update.
These recommendations apply to adults with a newly diagnosed lesion with a suspected or histopathologically proven glioblastoma (GBM). What are the optimal imaging techniques to be used in the management of a suspected glioblastoma (GBM), specifically: which imaging sequences are critical for most accurately identifying or diagnosing a GBM and distinguishing this tumor from other tumor types? Critical Imaging for the Identification and Diagnosis of Glioblastoma Level II: In patients with a suspected GBM, it is recommended that the minimum magnetic resonance imaging (MRI) exam should be an anatomic exam with both T2 weighted, FLAIR and pre- and post-gadolinium contrast enhanced T1 weighted imaging. The addition of diffusion and perfusion weighted MR imaging can assist in the assessment of suspected GBM for the purposes of distinguishing GBM from other tumor types. Computed tomography (CT) can provide additional information regarding calcification or hemorrhage and also can be useful for subjects who are unable to undergo MR imaging. At a minimum, these anatomic sequences can help identify a lesion as well as its location, and potential for surgical intervention. Improvement of diagnostic specificity with the addition of non-anatomic (physiologic imaging) to anatomic imaging Level II: One blinded prospective study and a significant number of case series support the addition of diffusion and perfusion weighted MR imaging in the assessment of suspected GBM, for the purposes of distinguishing GBM from other tumor types (e.g., primary CNS lymphoma or metastases). Level III: It is suggested that magnetic resonance spectroscopy (MRS) and nuclear medicine imaging (PET 18F-FDG and 11C-MET) be used to provide additional support for the diagnosis of GBM.


Effect of chronic guanethidine treatment on vascular norepinephrine content in the neonatal domestic pig.
Vinblastine, 6-hydroxydopamine, and antibody to nerve growth factor have been shown to destroy sympathetic neurons in neonatal rats. High doses of guanethidine produce permanent sympathectomies in the adult or newborn rat. Similar studies in the cat, rabbit, and hamster demonstrate that the sympathetic nervous systems are not destroyed with guanethidine. To produce permanent sympathectomy in a large animal model, guanethidine was administered to neonatal pigs. As with the cat, rabbit, and hamster, the sympathetic nervous system in the piglets was not destroyed by guanethidine.


Ringworm of the African hedgehog (Erinaceus albiventris) in the Ivory Coast due to Arthroderma benhamiae.
Isolates of the conidial state of Arthroderma behnhamiae (+ mating strain) were obtained from dry, scaly lesions on the ears of 3 hedgehogs in the Ivory Coast. This is a new record for this country.


Emulsion gels loaded with pancreatic lipase: Preparation from spontaneously made emulsions and assessment of the rheological, microscopic and cargo release properties.
Emulsion gels are solidified emulsions, which can be used for delivery of both hydrophilic and lipophilic substances. In this research, at first fish oil-in-water (O/W; 10% w/w) emulsions were prepared through the spontaneous emulsification technique. As emulsifier, a blend of the small-molecule surfactant tween 80, and either low-acyl (LaG) or high-acyl (HaG) gellan was used. For making fully stable (100% stability index) emulsions, a 10-fold higher concentration of LaG than HaG in the emulsion aqueous phase was required. The difference in gellan concentration resulted in a bigger mean drop size, as well as lesser consistency coefficient and yield stress for HaG emulsion than LaG emulsion. Subsequently, the fully stable HaG and LaG emulsions were gelled by CaCl2 addition. LaG emulsion gel was self-supporting and had a dense microstructure (as observed by electron microscopy), whereas HaG emulsion gel was not self-supporting. Loading lipase into the emulsions before ionotropic gelation did not lead to unacceptable acid values for fish oil during the emulsion gels storage. When the lipase-loaded fish O/W emulsion gels were immersed in an acid solution which imitated the gastric fluid (yet without digestive enzymes) oil droplets flocculated (as observed by confocal microscopy). The acid immersion also increased the dynamic moduli of the gels. Lipase was not released into the surrounding acid solution from LaG emulsion gel. A subsequent immersion within an alkaline solution imitating the small intestine fluid (yet without digestive enzymes) reduced the dynamic moduli of both kinds of emulsion gels. The alkaline immersion also caused extensive crack propagation in LaG emulsion gel network, which was found associated with diminished value of tan δ (G''/G') as an index of gel energy dissipation. Lipase was released form LaG emulsion gel into the alkaline solution, however, it took a remarkable period of time to begin.


Transfusion-acquired hepatitis E infection misdiagnosed as severe critical illness polyneuromyopathy in a heart transplant patient.
This is the case of a 56-year-old man who underwent heart transplantation. Within the first postoperative days, his respiratory and limb muscles weakened, which was attributed to critical illness polyneuromyopathy (CIPM). At day 70 post transplantation, he had increased liver enzyme levels and acute hepatitis E virus (HEV) infection was diagnosed. HEV RNA was found in the serum, stools, and cerebrospinal fluid. Results of further investigations suggested a possible HEV-related polyradiculoneuropathy. At transplantation, the patient was negative for immunoglobulin (Ig)G, IgM, and HEV RNA. A trace-back procedure identified the source of infection and concluded that HEV infection was contracted from blood transfusion 12 days prior to transplantation from an HEV RNA-positive donor. Tests of the organ donor for HEV were negative. Phylogenetic analysis revealed sequence homology between the HEV-3 strain of the patient and the HEV-3 strain of the blood donor. Despite ribavirin treatment, the patient died on day 153 post transplantation from multiorgan failure. In conclusion, patients with hepatitis or neuropathic illness who have received blood products should be screened for HEV.


A critical evaluation of Internet marketing of products that contain ephedra.
To evaluate information contained within Internet sites that advertise and market dietary supplements containing ephedra. We conducted an Internet search to identify Web sites advertising weight-loss supplements that contained ephedra. Between July 7 and July 18, 2002, 4 search engines were used by entering the term herbal weight loss. Outcome measures included disclosure of potential adverse effects of or contraindications to ephedra-containing supplements, disclosure of ephedra alkaloid dosage, and presence of misleading or incorrect information. Thirty-two products and advertisements were identified and systematically evaluated for deviance from truth-in-advertising standards. Of the 32 Web sites analyzed, 13 (41%) failed to disclose potential adverse effects or contraindications to supplement use. Seventeen (53%) did not reveal the dosage of ephedra alkaloids that was recommended. More importantly, 11 sites (34%) contained incorrect or misleading statements, some of which could directly result in serious harm to consumers. If dietary supplements containing ephedra are to continue to be marketed freely, substantial reform in advertising regulation and enforcement is warranted.


Impacts of thermal mismatches on chytrid fungus Batrachochytrium dendrobatidis prevalence are moderated by life stage, body size, elevation and latitude.
Global climate change is increasing the frequency of unpredictable weather conditions; however, it remains unclear how species-level and geographic factors, including body size and latitude, moderate impacts of unusually warm or cool temperatures on disease. Because larger and lower-latitude hosts generally have slower acclimation times than smaller and higher-latitude hosts, we hypothesised that their disease susceptibility increases under 'thermal mismatches' or differences between baseline climate and the temperature during surveying for disease. Here, we examined how thermal mismatches interact with body size, life stage, habitat, latitude, elevation, phylogeny and International Union for Conservation of Nature (IUCN) conservation status to predict infection prevalence of the chytrid fungus Batrachochytrium dendrobatidis (Bd) in a global analysis of 32 291 amphibian hosts. As hypothesised, we found that the susceptibility of larger hosts and hosts from lower latitudes to Bd was influenced by thermal mismatches. Furthermore, hosts of conservation concern were more susceptible than others following thermal mismatches, suggesting that thermal mismatches might have contributed to recent amphibian declines.


Effect of ethanol on hepatotoxicity and hepatic DNA-binding of aflatoxin B1 in rats.
The hepatocarcinogen aflatoxin B1 is converted to reactive metabolites that bind covalently to cellular macromolecules. These metabolites may also react with glutathione, resulting in the formation of glutathione conjugates and detoxication of the reactive metabolite. When rats were pretreated with ethanol by gastric intubation at a dose of 100 mmol/kg, 6 hr (the time of maximal GSH depletion) before the administration of aflatoxin B1, the covalent binding of 8,9-epoxide-aflatoxin B1 to DNA in vivo was increased by 47% and the hepatotoxicity was also potentiated. However, the covalent binding was not increased by pretreatment with ethanol 18 hr (time with approximately normal GSH levels) before administration of the toxin, and no potentiation of hepatotoxicity was observed. Pretreatment with a non-toxic dose of ethanol had no effects on the activity of glutathione S-transferase and glutathione peroxidase. These results suggest that the depletion of GSH and the increased formation of DNA-adduct from the liver constitute an important mechanism for the potentiation of aflatoxin B1-induced hepatotoxicity by ethanol.


Extracellular stimulation with human "noisy" electromyographic patterns facilitates myotube activity.
Electrical stimulation (ES) of skeletal muscle partially mimics the benefits of physical activity. However, the stimulation protocols applied clinically to date, often cause unpleasant symptoms and muscle fatigue. Here, we compared the efficiency of a "noisy" stimulus waveform derived from human electromyographic (EMG) muscle patterns, with stereotyped 45 and 1 Hz electrical stimulations applied to mouse myotubes in vitro. Human gastrocnemius medialis electromyograms recorded from volunteers during real locomotor activity were used as a template for a noisy stimulation, called EMGstim. The stimulus-induced electrical activity, intracellular Ca(2+) dynamics and mechanical twitches in the myotubes were assessed using whole-cell perforated patch-clamp, Ca(2+) imaging and optical visualization techniques. EMGstim was more efficient in inducing myotube cell firing, [Ca(2+)]i changes and contractions compared with more conventional electrical stimulation. Its stimulation strength was also much lower than the minimum required to induce contractions via stereotyped stimulation protocols. We conclude that muscle cells in vitro can be more efficiently depolarized using the "noisy" stochastic stimulation pattern, EMGstim, a finding that suggests a way to favor a higher level of electrical activity in a larger number of cells.


Development of American College of Rheumatology Quality Measures for Systemic Lupus Erythematosus: A Modified Delphi Process With Rheumatology Informatics System for Effectiveness (RISE) Registry Data Review.
We aimed to develop readily measurable digital quality measure statements for clinical care in systemic lupus erythematosus (SLE) using a multistep process guided by consensus methods. Using a modified Delphi process, an American College of Rheumatology (ACR) workgroup of SLE experts reviewed all North American and European guidelines from 2000 to 2020 on treatment, monitoring, and phenotyping of patients with lupus. Workgroup members extracted quality constructs from guidelines, rated these by importance and feasibility, and generated evidence-based quality measure statements. The ACR Rheumatology Informatics System for Effectiveness (RISE) Registry was queried for measurement data availability. In 3 consecutive Delphi sessions, a multidisciplinary Delphi panel voted on the importance and feasibility of each statement. Proposed measures with consensus on feasibility and importance were ranked to identify the top 3 measures. Review of guidelines and distillation of 57 quality constructs resulted in 15 quality measure statements. Among these, 5 met high consensus for importance and feasibility, including 2 on treatment and 3 on laboratory monitoring measures. The 3 highest-ranked statements were recommended for further measure specification as SLE digital quality measures: 1) hydroxychloroquine use, 2) limiting glucocorticoid use >7.5 mg/day to <6 months, and 3) end-organ monitoring of kidney function and urine protein excretion at least every 6 months. The Delphi process selected 3 quality measures for SLE care on hydroxychloroquine, glucocorticoid reduction, and kidney monitoring. Next, measures will undergo specification and validity testing in RISE and US rheumatology practices as the foundation for national implementation and use in quality improvement programs.


Electron microscopic study of cardiac ganglia in human fetuses.
The early events in the development of the heart ganglia and nerves in human fetuses ranging from 5 to 12 weeks of gestation age were studied by transmission electron microscopy. The first neuroblasts in the atrial mesenchyme differ from surrounding cells in the presence of short cisternae of rough endoplasmic reticulum and in the absence of glycogen particles. The most valuable criterion for identification of neuroblasts is the presence of contacts with preganglionic nerve terminals. Only ganglia composed of compact aggregations of neuronal cells and nerve terminals have a complete glial sheath. The first signs of synapse formation were seen in 5-week-old fetuses; well developed synapses with many synaptic vesicles were found from the 8th week of gestation onwards. These were predominantly axodendritic synapses. It is proposed that synaptogenesis begins with the appearance of osmiophilic zones at the sites of interneuronal contacts, then synaptic vesicles move by axonal transport to reach the preformed specialized junctions and stop in axonal presynaptic varicosities.


Enzyme replacement therapy in Hurler syndrome after failure of hematopoietic transplant.
The most severe form of Mucopolysaccharosidosis type I (MPS-I), Hurler syndrome, presents with progressive respiratory, cardiac and musculoskeletal symptoms and cognitive deterioration. Treatment includes enzyme replacement therapy (ERT) and/or hematopoietic stem cell transplantation (HSCT). We describe the case of an 8-year old boy with MPS-I, homozygous for W402X, treated at 10 months of age with HSCT and after failure of the transplant, with ERT during 2 years showing good results, including a positive neuropsychological development.


Sedimentation-induced detachment of magnetite nanoparticles from microalgal flocs.
The objective of this study is to develop a simple, one-step approach to separate adsorbed Fe3O4 nanoparticles from microalgal flocs for further downstream processing. Using the wild-type strain of fresh-water algae Chlamydomonas reinhardtii, effective removal of nanoparticles from microalgal flocs by both centrifugal sedimentation (at 1500 or 2000g) and magnetic sedimentation (at 1500 Oe) is demonstrated. At the physiological pH of the solution (i.e., pH 7.0), where the electrostatic force between the nanoparticles and the microalgal cells is strongly attractive, larger separation force was achieved by simply increasing the density and viscosity of the solution to 1.065g/mL and 1.244cP, respectively. The method described here offers significant opportunity for purifying microalgal biomass after nanoparticle-flocculation-based harvesting and decreasing the cost of microalgal biotechnology. This may also find avenues in other applications that apply flocculation, such as algal biofilm formation in photobioreactors and wastewater treatment.


Cancer genetic testing in marginalized groups during an era of evolving healthcare reform.
The Affordable Care Act and subsequent reforms pose tradeoffs for racial-ethnic, rural, and sex-related groups in the United States experiencing disparities in BRCA1/2 genetic counseling and testing and colorectal cancer screening, calling for policy changes. A working group of the American Public Health Association Genomics Forum Policy Committee engaged in monthly meetings to examine ongoing literature and identify policy alternatives in the coverage of cancer genetic services for marginalized groups. 589 items were collected; 408 examined. Efforts continued from February 2015 through September 2020. African Americans and Latinos have shown 7-8 % drops in uninsured rates since the Exchanges opened. The ACA has increased BRCA1/2 test availability while several disparities remain, including by sex. Rural testing and screening utilization rates have improved. Medicaid expansion and the inclusion of Medicare in the ACA have resulted in mixed improvements in colorectal cancer screening rates in marginalized groups. Cancer genetic testing and screening to date have only partially benefited from healthcare reforms. Sensitivity to cost concerns and further monitoring of emerging data are needed. A reduction in disparities depends on the availability of private insurance, Medicaid and Medicare to the marginalized. Attention to value-based design and the way cancer benefits are translated into actual testing and screening are crucial. The findings suggest the need for further benefits-related health agency interpretation of and amendments to the ACA, continued Medicaid and innovative Medicare expansion, and incorporation of cancer services values-based considerations at several levels, aimed at reducing group disparities.


Validation of a Simple Quality-of-Life Score for Identification of Minimal and Prediction of Overt Hepatic Encephalopathy.
Minimal hepatic encephalopathy (MHE) is underdiagnosed because most clinics refrain from psychometric testing. Diagnostic activities need to go up so patients with MHE can get the treatment their condition requires. The sickness impact profile questionnaire for covert hepatic encephalopathy (SIPCHE) score is based on quality-of-life outcomes and has been proposed as a simple, patient-administered diagnostic score for grade 1 and MHE. Validate the SIPCHE for MHE identification and overt hepatic encephalopathy (OHE) prediction. 110 patients with liver cirrhosis (age 60 years, Model for End-Stage Liver Disease score of 11.4, 80% blue-collar) provided information for SIPCHE scoring: gender, age, and four SIP statements: "I do not maintain balance (physically)," "I act irritable or impatient with myself," "I am not doing any of the usual physical recreation or activities," and "I am eating much less than usual." MHE was diagnosed using an abnormal continuous reaction time test and/or portosystemic encephalopathy syndrome test score. Patients were followed for 2.7 years on average. SIPCHE score positivity had high sensitivity (82%) but low specificity (38%) for MHE detection. Patients with an abnormal SIPCHE had a higher incidence of OHE during follow-up (35% vs. 14%, P = 0.05). OHE prediction sensitivity was 87% and exclusion sensitivity was 85%. The patients with an abnormal SIPCHE had twice as many subsequent episodes of OHE, and despite their high mortality, also a higher risk. An abnormal SIPCHE had a high sensitivity and low specificity for MHE identification. An abnormal SIPCHE was associated with a more than doubled risk of OHE, even with death as a competing event. SIPCHE could be used as a high-sensitivity, low-cost, surrogate marker of MHE in clinics without availability of psychometric tests and allow more patients to benefit from anti-MHE treatment.


Vitreous loss during phacoemulsification in glaucoma patients: long-term intraocular pressure control.
To assess the long-term effect of vitreous loss during phacoemulsification on intraocular pressure (IOP) control in glaucoma patients. Birmingham and Midland Eye Centre, City Hospital, Birmingham, United Kingdom. In this study, 26 glaucoma cases with vitreous loss during phacoemulsification were identified from operative room records. The cases were performed from January 1999 to December 2001. Minimum follow-up was 12 months. Postoperative IOP control in eyes with vitreous loss (Group 1) was assessed and compared with that in stable fellow eyes (Group 2), which served as controls. Another control group (Group 3) comprised patients with primary open-angle glaucoma who had successful phacoemulsification. Case notes of 22 patients from the same period fulfilled these criteria. The 3 groups were comparable in age, sex, laterality, ethnicity, mean IOP, and mean number of medications preoperatively. Twelve months after surgery, 43.2% in Group 1, 23.1% in Group 2 (P<.05) and 4.5% in Group 3 had significantly worse IOP; the differences between Group 1 and Groups 2 and 3 were statistically significant (P<.05). Intraocular pressure control was significantly better in Group 3 than in Group 2 (P<.05). Vitreous loss during cataract surgery in glaucoma patients adversely affected IOP control in the long term. Results suggest that uneventful cataract surgery in glaucoma patients improves IOP control.


GABAB modulation of neuronal activity related to visually guided movement in the monkey premotor cortex.
The effects of the gamma-aminobutyric acid-B (GABAB) receptor agonist baclofen and the GABAA receptor antagonist bicuculline methiodide on the activities of the same neurons in the premotor cortex of two monkeys while they performed a visually guided task with a GO or NO-GO response were investigated. While bicuculline methiodide induced excitatory responses in all of the task-related neurons examined, baclofen induced inhibitory responses in 75% of the examined neurons and excitatory responses in 25% of the neurons. The results indicate that, in the monkey premotor cortex, not only GABAA but also GABAB receptors are involved in GABAergic modulation of neuronal activities related to the visually guided behavioral performance.


Species diversification patterns in the Polynesian jumping spider genus Havaika Prószyński, 2001 (Araneae, Salticidae).
Hotspot archipelagoes provide exceptional models for the study of the evolutionary process, due to the effects of isolation and topographical diversity in inducing the formation of unique biotic assemblages. In this paper, we examine the evolutionary patterns exhibited by the jumping spider genus Havaika Prószyński, 2001 in the Polynesian islands of the Hawaiian and Marquesas chains. To date, systematic research on Havaika has been seriously limited by the poor taxonomic knowledge on the group, which was based on a handful of specimens that showed continuous variability and lacked clear-cut diagnostic characters. Here, we circumvent this problem by inferring a phylogeny based on DNA sequences of several fragments including both mitochondrial (protein coding cytochrome oxidase I, NAD1 dehydrogenase, ribosomal 16S, and tRNA leu) and nuclear (internal transcribed spacer 2) genes, and a statistical morphological analyses of a large sample of specimens. Results suggest that the Marquesan and Hawaiian Havaika may be the result of independent colonizations. Furthermore, data provide little support for the standard "progression rule" (evolution in the direction of older to younger islands) in Hawaiian Islands. This may be explained by a recent arrival of the group: age estimates of the different lineages suggest that Havaika colonized the Hawaiian Islands after most of the extant islands were already formed. The lack of clear-cut diagnostic characters among species may also be explained by the recent origin of the group since molecular data do not provide any evidence of hybridization among lineages. Quantitative morphological data coupled with the phylogenetic information allow us to reevaluate the current limitation of Havaika taxonomy. Molecular data support the existence of at least four different evolutionary lineages that are further morphologically diagnosable. However, genealogical relationships are better predicted by geographical affinity (i.e. island) than by morphological characters used in the original descriptions of the species. A pattern of size segregation linked to largely overlapping distributions of some of the species hints at a potential involvement of competition in generating morphological diversity. This study contributes to our understanding on the origin and shaping of the biodiversity of oceanic islands and sets the stage for more detailed studies on particular aspects of these previously overlooked spiders.


Temporal and microclimatic partitioning of the floral resources of Justicia aurea amongst a concourse of pollen vectors and nectar robbers.
The flowers of Justicia aurea, morphologically characteristic of ornithophily, attracted a diverse array of foragers where they occurred as a dense stand in the tropical forests at La Selva, Costa Rica, and so provided an arena for this study of competition and coexistence. Two hummingbird species (Phaethornis superciliosus and Campylopterus hemileucurus) visited the flowers legally early in the morning, and defended the nectar resource; a third smaller bird (P. longuemareus) foraged for nectar throughout the day but collected it illegally by piercing the corollas. In addition, nectar was harvested illegally by four species of stingless bee (Trigona) and by ants, creating a further drain on the limited floral resources.Consideration of the diurnal patterns of foraging activities in combination with a spatial axis (defined here in terms of microclimate and insolation) nevertheless showed a good separation of flower visits for the different nectarivores. Hummingbirds visited flowers in zones where the reward was highest, while insects foraged to minimise their energetic costs; each of these factors could in turn be related to microclimatic considerations. A comprehensive scheme of resource utilisation could therefore be extracted from the field observations and interpreted in these terms. The limited area of niche overlap thus revealed corresponded closely with the situations where direct interference competition was observed, between hummingbird species or between bees and aggressive ants.