bluuebunny/crossref_metadata_2025 huggingface dataset:
Distribution and expression of transgene green fluorescent protein in mice survived up to four weeks following in utero gene therapys
Authors: Yang P T , Huang L , Jia W W , Skarsgard E D , Sy C
Published: May 2017

Gene replacement offers a potential cure for degenerative disorders caused by a single gene deletion or mutation. Diagnoses of monogenic disorders in the fetus enable prenatal gene replacement which may be beneficial from the perspectives of host inflammatory/immune response, efficacy and disease prevention.

Purpose: To evaluate the distribution and expression of reporter gene green fluorescent protein (GFP) in the tissues of mice survived up to one month following in utero gene therapy.

Methods: Vesicular Stomatitis Virus-G (VSV-G) pseudotyped lentiviral (LV) vector containing GFP was prepared via triple plasmid co-transfection. Time-mated CD-1 mice underwent individual amniotic sac injection with either 1x106 LV particles or saline (controls) on gestational day 16 (term=21d), and were allowed to undergo spontaneous parturition. Pups were sacrificed on postnatal days 0, 7, 21 and 28, and neonatal and maternal tissues were analyzed for GFP transgene (by DNA polymerase chain reaction; PCR), and transgene expression by quantitative reverse transcriptase (QRT) PCR and immunohistochemistry (IHC).

Results: We observed selective transduction of neonatal tissues (trachea, lung, liver, heart, kidney, spleen, intestine, skeletal muscle), in pups undergoing in utero transfection with LV-GFP. Maternal tissues did not contain transgene despite exposure during amniotic injection. Although the numbers of pups analyzed at each postnatal time point was small, we observed variable persistence of GFP expression that appeared to be tissue specific (with persistent expression noted in intestine of 4 week old pups).

Conclusions: Neonatal tissue transfection occurs in a variety of tissues following amniotic injection with LV-GFP in this murine model of in utero gene therapy. Transgene persistence and expression patterns observed over the first 4 weeks of life may reflect tissue-specific genomic insertion of transgene that favors persistent transcription in select tissues.
Diagnosis and treatment of Paget’s disease of bone
Authors: Josse Robert G , Hanley David A , Kendler David , Ste Marie Louis-Georges , Adachi Jonathan D , Brown Jacques
Published: May 2017

Paget’s disease of bone (PDB) is a metabolic bone disease characterized by increased bone resorption followed by excessive unregulated bone formation. This results in weakened, deformed bones of increased mass in which the collagen fibres assume a haphazard irregular mosaic pattern instead of the normal parallel symmetry. PDB rarely occurs before middle age and its prevalence increases steadily with age. The overall prevalence in Caucasians is approximately 3%; although it appears to be declining. There is a geographic variation in prevalence, with highest rates found in the UK. PDB affects both men and women, with a slight predominance in men. PDB may be asymptomatic or symptomatic, depending on the bones involved; the most common symptom is pain in the affected bone. While its aetiology remains elusive, genetic factors and environmental influences are implicated. In 2002, guidelines for PDB management were developed in Great Britain and have gained worldwide acceptance. In this position paper, an Expert Panel of Canadian endocrinologists and rheumatologists examines current evidence on the diagnosis and treatment of PDB to provide Canadian recommendations. In general, diagnosis may be confirmed both by X-ray and by the biochemical marker serum alkaline phosphatase, which is elevated in 85% of individuals with untreated active PDB. Treatment is indicated for all patients with symptoms and for asymptomatic patients with active PDB in areas of the skeleton with the potential to produce complications of clinical importance. The Panel recommends treating PDB with bisphosphonates that have demonstrated superior efficacy and remission rates.
The significance of Toll-like receptor 4 (TLR4) expression in patients with chronic hepatitis B
Authors: Wei Xiu-Qing , Guo Yun-Wei , Liu Jia-Jun , Wen Zhuo-Fu , Yang Shao-Ji , Yao Ji-Lu
Published: May 2017

Purpose: To investigate the importance of Toll-like receptor 4 (TLR4) expression on hepatocytes obtained from Chronic Hepatitis B patients as well as on hepatocellular carcinoma HepG2 and HepG2.2.15 cell lines.

Methods: Expression of TLR4 in liver tissues was determined by immunohistochemistry in 75 patients with CHB and in10 healthy controls. The protein and mRNA 1eve1s of TLR4 in hepatocellular carcinoma HepG2 and HepG2.2.15 cells were measured by flow cytometry (FCM) and real-time quantitative PCR (RQ-PCR), and endotoxin triggered TNF-? secretion in HepG2 and HepG2.2.15 cells was evaluated by ELISA.

Results: TLR4 expressed mainly in the cytoplasm and some on cell membrane in hepatocytes. The staining scores of TLR4 expression in the liver tissues of patients with CHB were significantly higher than that of healthy controls. The liver tissues from patients with severe CHB expressed higher level of TLR4 than those from patients with mild CHB. Furthermore, the staining scores of TLR4 expression in the liver tissues of patients with CHB were positively correlated with the grading scores. Our results also showed that the mean fluorescence intensity and TNF-? secretion induced by endotoxin as well as the protein and mRNA 1eve1s of TLR4 in HepG2.2.15 cells were all significantly higher than those in HepG2 cells.

Conclusion: TLR4 was up-regulated in the hepatocytes in patients with CHB. This indicates a potentially important interaction between TLR4 expression and the pathogenesis of CHB.
MOTIVATION AND PLEASURE IN THE SCHIZOPHRENIA SPECTRUM–FINDINGS FROM THE PRODROME
Authors: Foussias G , Remington G , Mizrahi R
Published: May 2017

Background: Schizophreniais a chronic and debilitating illness that affects approximately one percent of the population. The symptoms of schizophrenia are typically thought of in separate domains, including positive symptoms (hallucinations and delusions), negative symptoms (diminished emotional expression and amotivation), and cognitive deficits. Importantly, the negative symptoms have been consistently found to adversely influence functional outcomes, in particular due to markedamotivation.^1 There have been suggestions that these individuals also experience deficits in the experience of pleasure, especially in their capacity to anticipate pleasure.^2 However, such investigations have not included the examination of these symptoms in those in the prodromal phase ofthis illness, a time that holds promise for early intervention and altering thecourse of schizophrenia.^3

Methods: In an effort to examine deficits in motivation and pleasure in the prodromal phase of schizophrenia, we have used an experience sampling method to assess “in the moment” motivation and pleasure in individuals at high risk of developing schizophrenia and healthy controls. Subjects completed baseline assessments including evaluation of their positive and negative symptoms. Subsequently, through the use of a personal digital assistant, subjects rated their motivation and experience of consummatory and anticipatory pleasure in their daily lives, multiple times over the course of four days.

Results and Conclusions: Preliminary data will be presented, as well as the importance of these findings in the context of understanding the underlying pathobiology of this illness, and guiding our search for effective treatments to improvefunctional outcomes in schizophrenia.

References:

1. Sayers SL, Curran PJ, Mueser KT. Psychol Assessment 1996;8:269-80.

2. Gard DE, Kring AM, Gard GM, et al.. Schizophr Res 2007;93:253-60.
COPY NUMBER VARIATION IN METABOLIC SYNDROME
Authors: Lanktree M , Hegele R A
Published: May 2017

Metabolic syndrome (MetS) is defined by the presence of abdominal obesity, hypertension, dysglycemia, and dyslipidemia. Many mutations have been discovered that cause rare monogenic components of metabolic syndrome, and association studies have linked common variants with increased risk of MetS and its components. Despite successes in identifying genetic contributors to metabolic syndrome, unexplained heritability exists and copy number variation (CNV) could be responsible for a portion of this variation. As observed with single nucleotide changes, it is likely that both rare and common CNVs will contribute to MetS disease susceptibility. Recent efforts to map CNVs in control populations have given insight into their size, frequency and distribution. However, despite being observed in controls, the reported CNVs could still modulate susceptibility for late-onset complex traitsor produce subtle metabolic phenotypes. Here we examine the overlap between CNVs found in control datasets and genes with functional hypotheses or evidence of previous association to MetS. Secondly, we present the results and methodology of a search for a rare CNV in a high-penetrance Mendelian disorder, namely familial partial lipodystrophy. As methods to identify CNVs increase in precision and accuracy, the prospect of identifying their role in both rare Mendelian and common complex diseases is exciting.
CORTTICAL AREA 5 IS NECESSARY FOR LONG-LASTING MEMORIES OF OBSTACLES ENCOUNTERED BY WALKKING CATS
Authors: McVea David , Pearson P KG
Published: May 2017

Vision is used by walking animals to ensure stable foot placements and to avoid obstacles in their path. Visual input is not used directly when walking over normal terrain, however. Instead, walking animals look three or four steps ahead of their current position and use short-term memory to remember pertinent objects for use at the appropriate time. In cluttered terrain, humans may rely on increased visual input, guiding foot movement directly when needed. In quadrupeds, the hindlegs can never beguided by visual input directly, suggesting a specialized memory system mightbe used during walking in cluttered terrain. We have previously reported evidence in support of this hypothesis, showing that cats that have stepped over an object with their forelegs and stopped remember its position for much longer (for up to ten minutes) than if they are stopped prior to stepping over it.

In this presentation, I present research that tests the hypothesis that neural signals related to the movement of the forelegs are essential for the generation of this unique visual memory. By placing small lesions in cortical area 5, we disrupted the normal integration of sensory and motor signals in the parietal cortex. Cats with these lesions lost the long-lasting memory of straddled objects, and remembered these objects for no longer than if they had not stepped over them. The results support our hypothesis that a uniquely long-lasting memory system, initiated by neural signals related to foreleg stepping, is responsible for guiding the hindlegs of walking cats over obstacles.

Funding from Canadian Institutes of Health Research and Alberta Heritage Foundation for Medical Research gratefully acknowledged.
THE DYNACTOME: INVESTIGATING DYNAMIC KINASE NETWORKS IN CELLULAR DECISION-MAKING
Authors: So Jonathan , Elder Kelly , Dai Anna , Jorgensen Claus , Linding Rune , Colwill Karen , Pawson Tony
Published: May 2017

Networks of kinases play a role in the transmission and integration of signals from the membrane to the nucleus. We aim to elucidate kinase phosphorylation and interaction partners in these networks through the immuno-precipitation and mass spectrometric analysis of a representative set of 100 Flag-tagged kinases stably expressed in human colorectal cancer cells. The goal is to generate a comprehensive set of interactions and dynamic phosphorylation sites which correlate with cell phenotypes such as apoptosis and proliferation. The techniques of mass-spectrometry have allowed for the identification of proteins and their phosphorylation sites in complex samples. Various labeling methods such as iTRAQ has enabled the relative quantification of these sites as afunction of time (White et al. PNAS, 2007). However, kinases usually work in the context of particular signaling stimuli. We aim to characterize the role of these over-expressed kinases in the context of Trail-induced apoptosis. This isparticularly relevant to tumorigenesis in that many cancers are resistant to apoptosis and recombinant Trail therapies are currently undergoing clinical trials. We present assays to correlate the proliferative ability and sensitivity to apoptosis of various stable cell lines with kinase expression levels through flow cytometry. We also present efforts to trace downstream signaling through the monitoring of MAP kinase phosphorylation using a high-throughput bead array.
HUMAN REGULATORY T CELLS PREVENT ISLET ALLOGRAFT REJECTION
Authors: Wu Douglas C , Wieckiewicz Joanna , Wood Kathryn J
Published: May 2017

Background: Type 1 diabetes mellitus represents a significant burden on global healthcare. Pancreatic islet transplantation offers an effective means of controlling the disease, but shortage of donor tissue, graft thrombosis, and immunological rejection after transplantation remain obstacles that need to be overcome. Our aim was to assess the ability of ex vivo expanded human regulatory T cells (Treg) in modulating the rejection response against a human islet allograft in a clinically relevant model of human pancreatic islet transplantation.

Methods: We studied the rejection response against allogeneic human islets in acohort of 32 immunodeficient mice which had been reconstituted with a functional human immune system. Thirteen subjects were transplanted with human islets without further immunological modification; graft survival was compared with that of thirteen subjects treated additionally with human regulatory T cells. Six controls were given a human islet transplant, but not reconstituted with human immune cells to demonstrate the functionality of the islet graft in the absence of immunological rejection. Graft function was assessed with serial blood glucose measurements, immunohistochemistry,immunoflourescence, and flow cytometry.

Findings: Human islet allografts were rapidly rejected in subjects that did notreceive Treg. With Treg treatment, however, human islet allograft rejection was prevented (median survival time (MST) of > 45 days with Treg, as opposed to an MST of 23 days without Treg). Ex vivo expanded Treg homed to the lymphoid tissue draining the graft site where they suppressed the priming, activation, proliferation, and effector cytokine production of alloreactive T cells.

Interpretation: These findings in a clinically relevant model of human pancreatic islet transplantation demonstrate the ability of ex vivo expanded human Treg to attenuate acute islet allograft rejection, and provide further support for their use in cellular immunotherapy.
The number, scope and geographic distribution of clinical researchers in Canada
Authors: Raghavan Malathi , Sandham J Dean
Published: May 2017

Purpose: Despite international concerns about declining numbers of clinical researchers, the number of clinical researchers in Canada remains undocumented.

Methods: The number and geographic distribution of clinical researchers in Canada and the scope of their research activities were estimated using, as an indicator, the data on clinical research projects funded by the Canadian Institutes for Health Research (CIHR).

Results: Between fiscal years 1999-00 and 2006-07, 1,041 individual researchers—approximately 130 per year—were principal investigators (PIs) on clinical research grants. One hundred and 26 researchers received salary awards; 449 supervisors oversaw the clinical research activities of 230 fellows and 223 students with trainee awards. An additional 2,305 individuals served only as co-investigators on grants. Most (863 [83%]) PIs received funding for operating grants; 196 (19%) PIs received funding for randomized controlled trials. The institute of neurosciences, mental health and addiction funded the highest number of researchers (187 [18%] PIs, 40 [17%] fellows, and 73 [33%] students). Among provinces, Quebec led the nation with the highest number (45) of PIs per million population. Ontario had the highest number of clinical research fellows (10 per million population) while Quebec and Saskatchewan each hosted more students (11 per million).

Conclusion: The number of Canadian investigators with funding for clinical research from CIHR was low. Although the ideal ratio of clinical to basic research capacity is not known, the possibility that the gap between laboratory-based research and clinical research is larger in Canada than in the United States is discussed.
Non-alcoholic fatty liver disease, obesity and other illnesses
Authors: Tarantino Giovanni
Published: May 2017

The estimated prevalence of NonAlcoholic Fatty Liver Disease in the general population in western countries is about 30%, but it is higher among obese and diabetic people. It is likely that more sophisticated approaches are required to understand its pathogenesis and to develop drug targets. In the meantime, the range of associations between NAFLD and other illnesses broadens. Although association does not mean causation, the link between some diseases and NAFLD suggests a common mechanism.
Oxidative stress and erythropoietin response in altitude exposure
Authors: Huang Hsien-Hao , Han Chih-Ly , Yan Horng-Chin , Kao Woei-Yau , Tsai Chu-Dang , Yen David Hung-Tsang , Huang Chun-I , Chen Wei-Teing
Published: May 2017

Purpose: Oxidative stress and erythropoietin (EPO) levels are increased following high altitude exposure. We hypothesized that the altitude-oxidative stress and EPO response would be associated with the presence or absence of acute mountain sickness (AMS) in subjects exposed at high altitude.

Methods: The study enrolled 29 healthy volunteers exposed at altitudes without strenuous physical exercise. Oxidative stress was determined by the spectrophotometric measurement of the colour occurring during the reaction of malondialdehyde (MDA) with thiobarbituric acid (TBA) on blood samples. Ferritin and EPO were also measured simultaneously.

Results: During a rise in altitude at 2000 and 3000 m, there were no changes in plasma ferritin level in either of the 2 groups with or without AMS. In contrast, EPO increased at an altitude of 3000 m and after returning to sea level (28.2±2.7, 26.9±3.3 vs 12.2±1.4 and 17.1±1.6, P < 0.05, in group without AMS; 29.3±4.5, 22.8±2.7 vs 10.6±1.0 and 16.1±1.5, # P < 0.05, in group with AMS; compared with the baseline level and at the height of 2000 meters). At a height of 3000 m, plasma MDA level was elevated compared with that at the altitude of baseline and 2000 m in both groups of subjects with and without AMS (3.77±0.29 vs 1.14±0.17, and 1.64±0.22, P < 0.001, in subjects with AMS; 3.65±0.39 vs 1.71±0.21, and 1.73±0.21, P < 0.001, in subjects without AMS) . After returning to sea level, subjects without AMS had lower MDA oxidative stress compared with those with AMS (2.58±0.26 vs 3.51±0.24, P = 0.0223). Along with a rise in altitude, the oxidative stress in these both groups was not correlated with the changes in EPO (r2 = 0.0728, P = 0.1096).

Conclusion: High altitude-induced oxidative stress, detected by MDA assay, is not different between the two groups of subjects with and without AMS. Upon return to sea level, subjects without AMS had lower MDA oxidative stress burden and higher EPO level than those with AMS. Whether the subjects with altitude illness had delayed recovery from oxidative stress merits further investigation.
Terazosin treatment suppresses basic fibroblast growth factor expression in the rat ventral prostate
Authors: Mitropoulos Dionisios , Kyroudi-Voulgari Aspasia , Christelli Evangelia , Zervas Anastasios , Karayannacos Panagiotis
Published: May 2017

Purpose: Alpha1-adrenergic receptor antagonists may not act solely on smooth muscle contractility. We evaluated the in vivo effect of the alpha1 blocker, terazosin, on the expression of basic fibroblast growth factor (bFGF) in the rat ventral prostate.

Methods: Wistar rats were treated with terazosin (1.2 mg/kg body weight, po, every second day) for 120 days. The expression of bFGF was assessed immuno-histochemically in tissue sections and by Western blotting in whole tissue preparations.

Results: Terazosin treatment did not affect prostate weight or histomorphology. In the control group, epithelial and stromal cells demonstrated positive staining for the anti-bFGF antibody. In contrast, the same staining in terazosin-treated specimens was either absent or extremely weak. An analogous difference was observed among the corresponding immunoblots.

Conclusions: These findings implicate the reduction of bFGF expression by terazosin as a potential additional molecular mechanism of its action that may include alterations in peptide growth factor mediated prostate homeostasis.
Telomerase activity (TMA) in tumour and peritumoural tissues in a rat liver cancer model
Authors: Zhang Huo-Jun , Yang Ji-Jin , Tian Jian-Ming , Wang Pei-Jun , Shao Cheng-Wei , Zuo Chang-Jing , Zhang Shun-Min , Gupta Sanjay
Published: May 2017

Purpose: To study the levels of telomerase activity (TMA) in tumour and peritumoural tissues in a liver cancer model in rats, and to study the change in TMA expression over time.

Methods: Using the telomeric repeated amplification protocol (TRAP), TMA was measured in tumour tissue, peritumoural tissue and normal liver tissue of Walker-256 tumour-bearing rats at 4, 6 and 8 days after tumour implantation.

Results: TMA at day 4, 6 and 8 was 0.767±0.117, 0.768±0.118 and 0.774±0.111 in tumour tissue, 0.389±0.263, 0.492±0.253 and 0.584±0.239 in peritumoural tissue, and 0.231±0.022, 0.229±0.022 and 0.233±0.021 in normal liver tissue, respectively. TMA in tumour tissue was higher than that in peri-tumour and normal liver tissues at all time points of measurement (P < 0.05). The TMA levels in tumour tissue and normal liver tissue did not show any change over time. TMA level in the peritumoural tissue increased with time; TMA level in animals sacrificed at day 8 was higher than that seen in animals sacrificed at day 4 (P < 0.05).

Conclusion: TMA in walker-256 tumour-bearing rats was higher than that in normal and peritumoural tissues. TMA level in the peritumoural tissue increased with time suggesting that TMA activation in peritumoural tissue may be an important factor promoting tumour growth.
Comparison of pharmacokinetics of L-carnitine, Acetyl-L-carnitine and Propionyl-Lcarnitine after single oral administration of L-carnitine in healthy volunteers
Authors: Cao Yu , Wang Yun-xiao , Liu Cheng-juan , Wang Le-xin , Han Zhi-wu , Wang Chun-bo
Published: May 2017

Purpose: To investigate the pharmacokinetics of L-carnitine (LC) and its analogues, acetyl-L-carnitine (ALC) and propionyl -L-carnitine (PLC) in healthy volunteers after single L-carnitine administration.

Methods: Liquid L-carnitine (2.0 g) was administered orally as a single dose in 12 healthy subjects. Plasma and urine concentrations of L-carnitine, ALC and PLC were detected by HPLC.

Results: The maximum plasma concentration (Cmax) and area under the curve (AUC0-?) of L-carnitine was 84.7±25.2 ?mol·L-1·h and 2676.4±708.3 ?mol·L-1·h, respectively. The elimination half-life of L-carnitine and the time required to reach the Cmax (Tmax) was 60.3±15.0 and 3.4±0.46 h, respectively. The Cmax of ALC (12.9±5.5 ?mol·L-1) and PLC (5.08±3.08 ?mol·L-1) was lower than L-carnitine (P < 0.01), so as the AUC0-? (166.2±77.4 and 155.6±264.2?mol·L-1·h, respectively, P < 0.01). The half-life of ALC (35.9±28.9h) and PLC (25.7±30.3 h) was also shorter than L-carnitine (P < 0.01). The 24h accumulated urinary excretion of L-carnitine, ALC and PLC were 613.5±161.7, 368.3±134.8 and 61.3±37.8?mol, respectively.

Conclusion: L-carnitine has a greater maximum plasma concentration than ALC and PLC. L-carnitine also has a longer half-life than ALC and PLC. These data may have important implications in the designing of dosing regimens for L-carnitine or its analogues, such as ALC or PLC.
Keloid scarring, but not Dupuytren’s contracture, is associated with unexplained carotid atherosclerosis
Authors: Bhavsar Sankalp , Nimigan Andre , Hackam Daniel G , O'Gorman David B , Gan Bing Siang , Spence J David
Published: May 2017

Background: Atherosclerosis, a response to injury, may be thought of as scarring in the artery wall. TGF-? and associated signaling molecules have been implicated in the pathophysiology of keloid scarring, Dupuytren’s Contracture and atherosclerotic plaques in independent studies.

Purpose: To test the hypothesis that excess cutaneous scarring and Dupuytren’s contractures predispose independently to carotid atherosclerosis .

Methods: Among 1,747 patients with plaque measurements and complete data for multivariable regression analysis, 57 Caucasian patients had Dupuytren’s contractures and 12 had keloid scars. Carotid total plaque area (TPA) was measured by 2-Dimensional ultrasound.

Results: In linear multivariable regression analysis with coronary risk factors, keloid scars were associated with TPA (P= 0.018), but Dupuytren’s contractures were not. Patients with keloid scarring were younger (P < 0.0001), and more likely to be diabetic (P < 0.0001)

Conclusions: Keloid scarring is a clinical clue to excess atherosclerosis not explained by traditional risk factors. Such patients may benefit from therapy directed at targets related to signalling molecules common to both the process of keloid scarring and atherosclerosis. These findings suggest previously unexplored possibilities for the prevention and treatment of atherosclerosis. The differences between Dupuytren’s and keloid scars that may identify such targets are discussed.
Graves’ disease treated with thyroid arterial embolization
Authors: Zhao Wei , Gao Bu-Lang , Tian Min , Yi Gen-Fa , Yang Hui-Ying , Shen Li-Juan , Li Hong , Shen Jin
Published: May 2017

Purpose: To study pathological changes in the thyroid gland of patients with Graves’ disease (GD) treated with thyroid arterial embolization.

Methods: Thirty-seven patients with GD were treated through transcatheter thyroid arterial embolization. Of these patients, twenty-two had biopsy of the thyroid gland at different time points before and after the embolization for the study of pathology. Serum thyroid hormones, TSH and TRAb were also studied at these time points. Thyroid size was evaluated in all patients using color Doppler ultrasound or CT scan.

Results: Thyroid size decreased immediately or several days following embolization. Pathological study demonstrated mainly acute infarction and necrosis at 7 days post embolization. At 6 months, chronic inflammation and fibrous hyperplasia were the primary findings in the gland, and at 3 years following embolization, mesenchyma hyperplasia and follicle atrophy were primarily present in the embolized thyroid tissue. The thyroid hormones and TSH gradually resumed to normal range after embolization while TRAb decreased significantly.

Conclusion: Thyroid arterial embolization can cause GD thyroid gland a series of pathological changes of acute ischemia and necrosis and later, chronic inflammation, fibroplasia and atrophy to decrease secretion of thyroid. The pathological changes within the thyroid gland after embolization form the basis of thyroid arterial embolization in treating GD hyperthyroidism.
Predictors of time to remission and treatment failure in patients with Graves’ disease treated with propylthiouracil
Authors: Cinemre Hakan , Bilir Cemil , Gokosmanoglu Feyzi , Akdemir Nermin , Erdogmus Besir , Buyukkaya Ramazan
Published: May 2017

Purpose: Propylthiouracil is one of the thionamides used in the treatment of Graves’ disease. The drug has serious side effects and long-term treatment might be needed to achieve remission.

We designed this study to evaluate the clinical and thyroid Doppler characteristics that might predict time to remission and treatment failure in propylthiouracil treated Graves’ patients.

Methods: 26 patients, among 134 presenting to our university hospital outpatient clinic between Feb -July 2007 and with first time diagnosis of clinical thyroid dysfunction, were clinically and ultasonographically diagnosed with Graves’ disease. Doppler parameters, serum thyrotropin, free thyroxine and free triiodothyronine were measured at the beginning of the study and thyroid studies were repeated every 4 weeks until remission. Propylthiouracil 300 mg/day was started for each patient at the time of diagnosis and doses were titrated according to repeat thyroid studies. Patients were treated and followed up for 18 months.

Results: Treatment failure was associated with smoking (P = 0.001) and male gender (P= 0.037). Stepwise multiple regression analysis revealed that age, free thyroxine and superior thyroid artery flow rate were predictors of time to remission (P= 0.001, 0.002 and 0.003, respectively).

Conclusion: The time to remission in Graves patients treated with propylthiouracil can be predicted using age, serum free thyroxine and superior thyroid artery flow rate. This may help early consideration of alternative treatment for the patients requiring prolonged treatment for remission or for those who fail medical treatment. This would decrease unnecessary, long-term propylthiouracil exposure with its serious side effects.
Serum levels of p53 and cytochrome c in subjects with type 2 diabetes and impaired glucose tolerance
Authors: Dincer Yildiz , Himmetoglu Solen , Yalin Serap , Damci Taner , Ilkova Hasan , Akcay Tulay
Published: May 2017

Purpose: To examine apoptotic markers in serum of subjects with diabetes and impaired glucose tolerance (IGT). Serum levels of p53 and cytochrome c, regulator molecules for apoptosis, were measured in subjects with type 2 diabetes, subjects with IGT and healthy controls.

Methods: Forty one subjects with type 2 diabetes, 27 with IGT and 27 healthy volunteers were included in the study. Serum level of cytochrome c and p53 were measured with competitive ELISA.

Results: Serum levels of p53 were lower in the group of subjects with type 2 diabetes (085±0.39 U/ml) than in controls (1.09±0.49 U/ml) (P < 0.05) and in the subjects with IGT (0.98±0.37 U/ml) (P < 0.05). There was no significant difference between the group with IGT and controls. Also, there was no difference for serum level of cytochrome c among the groups. In the group of subjects with type 2 diabetes, serum level of cytochrome c was mildly correlated with HbA1c (r:0.39, P < 0.05).

Conclusion: The present study shows that the serum level of p53 is lower in the patients with type 2 diabetes than in controls or in subjects with IGT. No difference was seen among the the groups for the serum level of cytochrome c.
Adiponectin has different mechanisms in type 1 and type 2 diabetes with C-peptide link
Authors: Ljubic Spomenka , Boras Jozo , Jazbec Anamarija , Lovrencic Marijana Vucic , Vidjak Vinko , Erzen Dubravka Jurisic , Mileta Dean
Published: May 2017

Purpose: Adiponectin (ApN) is considered to be responsible for reduction of inflammation and is known to be included in lipid metabolism. This study was designed to assess the role of adiponectin in patients with type 1 and type 2 diabetes and to determine parameters important in the prediction of adiponectin.

Methods: Adiponectin, high sensitive C-reactive protein, fibrinogen, homocysteine, C-peptide, and lipid panel in addition to clinical and laboratory parameters important for the definition of diabetes, obesity and the metabolic syndrome were measured in 118 patients.

Results: The best model (R2=0.989) for predicting adiponectin in type 1 diabetes included fibrinogen, white blood cell count, uric acid and triglycerides. In type 2 diabetes the best model (R2=0.751) included C-peptide, white blood cell count, systolic blood pressure, fasting blood glucose, glycated hemoglobin and high-density lipoprotein cholesterol. ANOVA showed among-group differences in adiponectin (P=0.028), body mass index (P < 0.001), fasting blood glucose (P < 0.001) and high-density lipoprotein cholesterol (P =0.012) according to the type of diabetes. Between-group differences were also observed in adiponectin (P =0.033) and high-density lipoprotein cholesterol (P =0.009) according to sex. Adiponectin correlated (P < 0.05) with body mass index, C-peptide, pulse pressure and high-density lipoprotein cholesterol.

Conclusion: Adiponectin levels were higher in type 1 diabetes. The association between C-peptide and adiponectin is probably one of the reasons for their different respective levels in different types of diabetes. Interrelations between adiponectin and inflammation, dyslipidemia, C-peptide levels and sex appear to be important for complex adiponectin modulation and action.
Potential role of matrix metalloproteinase-2,-9 and tissue inhibitors of metalloproteinase-1,-2 in exudative pleural effusions
Authors: Vatansever S , Gelisgen R , Uzun H , Yurt S , Kosar F
Published: May 2017

Purpose: To investigate diagnostic values of pleural fluid matrix metalloproteinase-2 (MMP-2), MMP-9, tissue inhibitors of metalloproteinase-1 (TIMP-1) and TIMP-2 measurements in tuberculous pleurisy(TP) and malignat pleurisy (MP).

Methods: The study included 24 patients with TP, 22 patients with MP and 15 patients with pleural effusion of non-tuberculous and non-malignant origin as controls. MMP-2,-9 and TIMP-1,-2 levels in pleural fluid were measured by ELISA method.

Results: Pleural fluid MMP-2 and MMP-9 levels were higher (P < 0.001, P < 0.001, respectively) in TP than in MP and controls. MP patients have higher pleural fluid MMP-2 and MMP-9 levels (P < 0.01, P < 0.05, respectively) than controls. Pleural fluid TIMP-2 levels were higher (P < 0.01 and P < 0.001, respectively) in MP than in TP and controls. Pleural fluid MMP-9 levels were negatively correlated with pleural fluid TIMP-2 levels (r: 0.464, P=0.029) in patients with MP.

Conclusions: Determination of TIMP-2 in pleural fluid may contribute to differentiate TP from MP. These results suggest that overproduction of MMP-9 and TIMP-2 is associated with accumulation of the pleural effusion in malignancy. Further studies with a greater number of patients are needed to confirm this hypothesis.
CSCI-2009-2010 Call for Award Submissions
Authors: CSCI _
Published: May 2017

CSCI 2009-2010 AWARDS CALL FOR SUBMISSIONS

The Canadian Society for Clinical Investigation is pleased to request submissions for the following awards:

Deadline November 26, 2009:

Dr. Mel Silverman Distinguished Service Award CSCI Distinguished Scientist Award CSCI Joe Doupe Young Investigator Award

Deadline January 29, 2010:

CSCI/RCPSC Henry Friesen Award Canadian Research Awards for Specialty Residents (Medicine, Surgery)

Deadline July 5, 2010:

CSCI/CIHR Resident Research Prize

More information will follow on the CSCI website at www.csci-scrc.org
Angiogenic study in Graves’ disease treated with thyroid arterial embolization
Authors: Zhao Wei , Gao Bu-Lang , Liu Zhi-Yong , Yi Gen-Fa , Shen Li-Juan , Yang Hui-Ying , Li Hong , Song Dian-Ping , Jiang Yong-Neng , Hu Ji-Hong , Luo Gang
Published: May 2017

Purpose: To investigate angiogenesis in the thyroid of Graves’ disease (GD) treated with thyroid arterial embolization through analysis of vascular endothelial growth factor (VEGF), basic fibroblast growth factor (bFGF) and microvessel density (MVD).

Materials and methods: Forty-two GD patients were treated with thyroid arterial embolization and followed up for 1-68 months after embolization. Before embolization and at 7 days, 3, 6, 12, 36 and 48 months following embolization, TT3, TT4, FT3, FT4, TSH and thyroid stimulating antibody (TSAb) were tested respectively. Thyroid biopsy was performed under the guidance of computed tomography for immunohistochemical staining of VEGF and bFGF, and MVD within the thyroid gland was marked by CD34.

Results: VEGF and bFGF were mostly expressed in the cytoplasm and on the cell membrane. The expression of VEGF was increased (P < 0.05) at ?6 months compared with before embolization and decreased (P < 0.05) at ?1 year compared with either at ?6 months or before embolization. The expression of bFGF was not statistically different at ?6 months compared with before embolization but was decreased (P < 0.05) at ?1 year compared with either at ?6 months or before embolization. Thyroid MVD marked by CD34 had similar changes to those of the VEGF expression after embolization. There was a positive correlation between VEGF and bFGF (P < 0.05) and between VEGF or bFGF and MVD (P < 0.05). Thyroid hormones mostly returned to normal and TSAb was decreased in longer follow-up.

Conclusion: Thyroid arterial embolization can decrease the expression of VEGF, bFGF and MVD. Consequently, angiogenesis within the GD thyroid will be decreased in the long term after embolization and may serve as the basis for reduced thyroid size and function.
Cigarette smoking and chronic low back pain in the adult population
Authors: Alkherayf Fahad , Agbi Charles
Published: May 2017

Purpose: Chronic low back pain (LBP) is one of the main causes of disability in the community. Although there have been studies suggesting an association between smoking and LBP, these studies were limited by the small numbers of patients, and they did not control for confounders. The objective of this study was to determine whether cigarette smoking is associated with an increased risk of chronic LBP among adults.

Methods: Using Canadian Community Health Survey (cycle 3.1) data, 73,507 Canadians aged 20 to 59 yr were identified. Self-reported chronic LBP status, smoking habits, sex, age, height, weight, level of activity and level of education were identified as well. Back pain secondary to fibromyalgia was excluded. Multivariate logistic regression analysis was used to detect effect modification and to adjust for covariates. Design effects associated with complex survey design were taken into consideration.

Results: The prevalence of chronic LBP was 23.3% in daily smokers and only 15.7% in non-smokers. Age and sex were found to be effect modifiers (P < 0.0001), and the relationship between smoking and chronic LBP risk was dependent on sex and age. The association between daily smoking and the risk of chronic LBP was stronger among younger individuals. Occasional smoking slightly increased the odds of having chronic LBP.

Conclusion: Daily smoking increases the risk of LBP among young adults, and this effect seems to be dose-dependent. Back pain treatment programs may benefit from integrating smoking habit modification. Further research is required to develop effective prevention strategies.
Relationship between hemoglobin and CD40 ligand in prediabetes
Authors: Kutlu Mustafa , Sonmez Alper , Genc Halil , Erdem Gokhan , Tapan Serkan , Celebi Gurkan , Haymana Cem , Taslipinar Abdullah , Uckaya Gokhan , Erbil Mehmet Kemal
Published: May 2017

Purpose: Hemoglobin (Hb) regulates the endothelial function by modulating the bio-availability of NO at the tissue level. A significant direct relationship is present between the Hb levels and endothelial functions in patients with Type 2 diabetes. Testing whether this association also exists in subjects with prediabetes is important because prediabetes is associated with an increased risk of cardiovascular disease and mortality. Therefore, we investigated the association of Hb both with the classical cardiac risk factors and the markers for endothelial dysfunction and inflammation, in subjects with impaired glucose tolerance (IGT).

Methods: We enrolled 69 normotensive, and cardiovascular events free subjects with IGT (M=40, age=45.50±6.8 yr). Plasma insulin, hsCRP, soluble CD40L, vonWillebrand factor, p-selectin levels were measured. The parameters given according to the higher and lower median Hb values of the subjects were compared.

Results: Subjects with the higher Hb levels exhibited lower HDL-C (46.68±10.8 mg/dl vs 51.5±8.9mg/dl; P=0.04) and higher systolic (122.57±6.2mmHg vs 116.17±7.4mmHg; p < 0.001) and diastolic (79.14±3.73mmHg vs 75.58±6.1mmHg; P=0.005) blood pressures and sCD40L (7.9±3.8ng/ml vs 6.07±2.1 ng/ml; P=0.02) levels. Hb levels were correlated to the HDL cholesterol, sCD40L, systolic and diastolic blood pressures and waist circumference (r=-0.28, P=0.02; r=0.29, P=0.02; r=0.53, P < 0.001; r=0.41, P=0.001; r=0.42, P < 0.001 respectively). According to the multiple logistic regression analysis, Hb was the determinant of sCD40L levels (?=0.437, P < 0.001).

Conclusion: These results indicate that there may be a link with higher Hb values and cardiovascular risk factors in patients with IGT. Further investigation is warranted to understand the clinical implications of these findings in subjects with prediabetes.
METHOD VALIDATION STUDY TO EVALUATE THE ANALYTICAL PERFORMANCE OF THE STAT–SITE METER FOR THE MEASURMENT OF SERUM BETA-HYDROXYBUTRATE
Authors: Elsharif A , Don-Wauchope A
Published: May 2017

Objectives: To evaluate the analytical performance of the STAT-Site meter (Stanbio laboratory USA) for the measurement of serum beta-hydroxybutyrate (? OH-B) concentration.

Methods: The precision was evaluated using two levels of quality control materials (low and high). Within run and between run precision studies were performed. CVs (coefficient of variation) compared against accepted standards. Fifty-one leftover patient samples with previously reported ? OH-B concentrations were used for the method comparison. The accuracy was evaluated and compared to our current laboratory method, (Wako Autokit 3-HB, Wako chemical USA). Slope, intercept, correlation coefficient, Deming regression and paired t-test were calculated using Analyse-it® software

Results: The meter showed reasonable precision for the measurement of ? OH-B with CV of 0.0% and 6% for within run precision study and 8.82% and 10.07% for between run precision study which are acceptable according to two CAP surveys. Further these are comparable to our current method CV and the manufacturer claimed CV’s. Deming regression analysis showed a linear relationship between the two methods: The slope of the regression equation was 0.98 (95% CI, 0.87-1.08); intercept 0.10 (95% CI 0.03-0.16) and correlation coefficient of 0.979.There was no bias detected by the visual inspection of the difference plot, this was confirmed by the calculation of the paired t-test (p = 0.19).

Conclusion: The STAT-Site meter is a practical, rapid method with wide analytical range that meets precision and accuracy criteria. The method is suitable for the use in a research context until it gets approval for clinical use.
ASSESSING THE FREQUENCY OF CA-125 MEASUREMENTS WITHIN HAMILTON HEALTH SCIENCES AND ST. JOSEPH’S HEALTHCARE
Authors: Nawara A , Hirte H , Kavsak P
Published: May 2017

Objective: To determined if the number and frequency of CA-125 measurements per patient reflect the 2008 NACB (National Academy of Clinical Biochemist) practice guidelines.

Methods: We collected data retrospectively on CA-125 over a period of one year as apart of our ongoing practice in monitoring the tumor markers for quality assurance, The number of CA-125 results per patient as well as the time interval between the 1st and 2nd measurements was noted. To enrich this population for the likelihood that the measurement of CA-125 was used for monitoring or for detection of recurrence, we divided the population into patients from the Jurvainski Cancer Centre (JCC) and the rest of the sites.

Results: The JCC patients contributed the majority of CA-125 results, representing over 75% of all results (n= 3057 results from 998 patients), whereas the remainder of the sites only yielded 959 results from 920 patients. Further analysis of the JCC patients indicated 2 main subgroups: Group A – patients with 1 or 2 results (n=624 patients); Group B - patients with 3 or more results , Overall, less than 4% of patients at the JCC had a time interval between the 1st and 2nd specimens of less than 2 weeks.

Discussion: This initial analysis would indicate that physicians are ordering CA-125 in agreement with the NACB guidelines. To further improve compliance to the guidelines and to prevent subsequent measurements of CA-125 too close, we propose restricting CA-125 orders that are less than 14 days apart to only those that receive Biochemist’s approval.
The role of C1GALT1C1 in lipopolysaccharide-induced IgA1 aberrant O-glycosylation in IgA nephropathy
Authors: Xie Lin-Shen , Qin Wei , Fan Jun-Ming , Huang Jun , Xie Xi-Sheng , Li Zi
Published: May 2017

Purpose: IgA1 aberrant O-glycosylation is one of the main pathogenetic features of IgA nephropathy (IgAN). This study attempted to determine the role of C1GALT1C1 in aberrant IgA1 O-glycosylation induced by lipopolysaccharide (LPS) and identify potential therapeutic targets in IgAN.

Methods: Lymphocytes isolated from 22 patients with IgAN and 17 normal controls were cultured for 3 to 7 days with or without LPS and 5-azacytidine (5-AZA). Expression levels of C1GALT1C1 mRNA and protein were measured by real-time PCR and Western blot analysis, respectively. Concentration of IgA1 and level of O-glycosylation were determined by ELISA and Vicia villosa (VV) lectin-binding assay. Correlation analysis was performed between the expression of C1GALT1C1 protein and IgA1 O-glycosylation.

Results: Lymphocytes from patients with IgAN secreted more IgA1 than that from normal controls after LPS stimulation (P=0.26, 0.002 and 0.005 on the 3rd, 5th and 7th day, respectively) which could be inhibited by 5-AZA (P=0.001, 0.025 and 0.001 on the 3rd, 5th and 7th day, respectively). Moreover, LPS stimulation could obviously inhibit C1GALT1C1 expression in patients with IgAN (decreased by 71%, 82% and 92% on the 3rd, 5th and 7th day, respectively; P < 0.001), and cause a significant decrease of IgA1 O-glycosylation compared with normal controls (P=0.004, 0.003 and 0.03 on the 3rd, 5th and 7th day, respectively). When 5-AZA was added, the level of C1GALT1C1 expression increased dramatically (1.98, 5.53 and 8.97 times on the 3rd, 5th and 7th day, respectively; P < 0.001) along with an increase of IgA1 O-glycosylation (P=0.295, 0.09 and 0.003 on the 3rd, 5th and 7th day, respectively). However, normal controls showed no significant change in C1GALT1C1 expression and IgA1 O-glycosylation after LPS stimulation (P > 0.05).

Conclusion: LPS induced IgA1 aberrant O-glycosylation and suppressed C1GALT1C1 expression in patients with IgAN. Upregulation of C1GALT1C1 expression by 5-AZA could reverse the IgA1 aberrant O-glycosylation. These results suggest that C1GALT1C1 may play a key role in the regulation of IgA1 O-glycosylation.