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1.
Diabet Med ; 35(7): 846-854, 2018 07.
Article in English | MEDLINE | ID: mdl-29577410

ABSTRACT

AIMS: To assess adult diabetes care providers' current transition practices, knowledge about transition care, and perceived barriers to implementation of best practices in transition care for emerging adults with Type 1 diabetes mellitus. METHODS: We administered a 38-item web-based survey to adult diabetes care providers identified through the Québec Endocrinologist Medical Association and Diabetes Québec. RESULTS: Fifty-three physicians responded (35%). Fewer than half of all respondents (46%) were familiar with the American Diabetes Association's transition care position statement. Approximately one-third of respondents reported a gap of >6 months between paediatric and adult diabetes care. Most (83%) believed communication with the paediatric team was adequate; however, only 56% reported receiving a medical summary and 2% a psychosocial summary from the paediatric provider. Respondents believed that the paediatric team should improve emerging adults' preparation for transition care by developing their self-management skills and improve teaching about the differences between paediatric and adult-oriented care. Only 31% had a system for identifying emerging adults lost to follow-up in adult care. Perceived barriers included difficulty accessing psychosocial services, emerging adults' lack of motivation, and inadequate transition preparation. Most (87%) were interested in having additional resources, including a self-care management tool and a registry to track those lost to follow-up. CONCLUSIONS: Our findings highlight the need to better engage adult care providers into transition care practices. Despite adult physicians' interest in transition care, implementation of transition care recommendations and resources in clinical care remains limited. Enhanced efforts are needed to improve access to mental health services within the adult healthcare setting.


Subject(s)
Attitude of Health Personnel , Communication , Diabetes Mellitus, Type 1/therapy , Endocrinology , Pediatrics , Transition to Adult Care , Adult , Cross-Sectional Studies , Female , Health Services Accessibility , Humans , Interprofessional Relations , Lost to Follow-Up , Male , Middle Aged , Motivation , Psychosocial Support Systems , Quebec , Self Care , Surveys and Questionnaires
2.
Curr Oncol ; 25(6): e597-e609, 2018 12.
Article in English | MEDLINE | ID: mdl-30607129

ABSTRACT

Background: Colorectal Cancer Canada, in partnership with a Scientific Advisory Committee, is developing a Canadian Patient Group Pathway to Accessing Cancer Clinical Trials ("Pathway"). A central element of the Pathway is presented here-namely, a set of recommendations and tools aimed at each stakeholder group. Methods: A summary of the peer-reviewed and grey literature informed discussions at a meeting, held in June 2017, in which a cross-section of stakeholders reached consensus on the potential roles of patient groups in the cancer clinical trials process, barriers to accessing cancer clinical trials, best practice models for patient-group integration, and a process for developing the Pathway. Canadian recommendations and tools were subsequently developed by a small working group and reviewed by the Scientific Advisory Committee. Results: The major output of the consensus conference was agreement that the Clinical Trials Transformation Initiative (ctti) model, successfully applied in the United States, could be adapted to create a Canadian Pathway. Two main differences between the Canadian and American cancer clinical research environments were highlighted: the effects of global decision-making and systems of regulatory and funding approvals. The working group modified the ctti model to incorporate those aspects and to reflect Canadian stakeholder organizations and how they currently interact with patient groups. Conclusions: Developing and implementing a Canadian Pathway that incorporates the concepts of multi-stakeholder collaboration and the inclusion of patient groups as equal partners is expected to generate significant benefits for all stakeholders. The next steps to bring forward a proposed Pathway will involve engaging the broader cancer research community. Clinical trial sponsors will be encouraged to adopt a Charter recognizing the importance of including patient groups, and to support the training of patient groups through an independent body to ensure quality research partners. Integration of patient groups into the process of developing "real world" evidence will be advanced by a further consensus meeting being organized by Colorectal Cancer Canada for 6-7 November 2018.


Subject(s)
Clinical Trials as Topic , Critical Pathways , Biomedical Research , Canada , Cross-Sectional Studies , Decision Making , Health Planning Guidelines , Humans , Models, Theoretical
3.
Theriogenology ; 86(6): 1482-1488, 2016 Oct 01.
Article in English | MEDLINE | ID: mdl-27341774

ABSTRACT

The objectives of this study were to determine (i) whether Chlamydia abortus would adhere to or penetrate the intact zona pellucida (ZP-intact) of early in vivo-derived caprine embryos, after in vitro infection; and (ii) the efficacy of the International Embryo Transfer Society (IETS) washing protocol for bovine embryos. Fifty-two ZP-intact embryos (8-16 cells), obtained from 14 donors were used in this experiment. The embryos were randomly divided into 12 batches. Nine batches (ZP-intact) of five embryos were incubated in a medium containing 4 × 10(7)Chlamydia/mL of AB7 strain. After incubation for 18 hours at 37 °C in an atmosphere of 5% CO2, the embryos were washed in batches in 10 successive baths of a phosphate buffer saline and 5% fetal calf serum solution in accordance with IETS guidelines. In parallel, three batches of ZP-intact embryos were used as controls by being subjected to similar procedures but without exposure to C. abortus. The 10 wash baths were collected separately and centrifuged for 1 hour at 13,000 × g. The washed embryos and the pellets of the 10 centrifuged wash baths were frozen at -20 °C before examination for evidence of C. abortus using polymerase chain reaction. C. abortus DNA was found in all of the infected batches of ZP-intact embryos (9/9) after 10 successive washes. It was also detected in the 10th wash fluid for seven batches of embryos, whereas for the two other batches, the last positive wash bath was the eighth and the ninth, respectively. In contrast, none of the embryos or their washing fluids in the control batches were DNA positive. These results report that C. abortus adheres to and/or penetrates the ZP of in vivo caprine embryos after in vitro infection, and that the standard washing protocol recommended by the IETS for bovine embryos, failed to remove it. The persistence of these bacteria after washing makes the embryo a potential means of transmission of the bacterium during embryo transfer from infected donor goats to healthy recipients and/or their offspring. Nevertheless, the detection of C. abortus DNA by polymerase chain reaction does not prove that the bacteria found was infectious. Further studies are required to investigate whether enzymatic and/or antibiotic treatment of caprine embryos infected by C. abortus would eliminate the bacteria from the ZP.


Subject(s)
Chlamydia Infections/veterinary , Chlamydia , Embryo Transfer/veterinary , Goat Diseases/embryology , Goat Diseases/microbiology , Animals , Chlamydia/genetics , Chlamydia/isolation & purification , Chlamydia Infections/transmission , DNA, Bacterial/analysis , Embryo, Mammalian/microbiology , Goats , Polymerase Chain Reaction/veterinary , Zona Pellucida/microbiology
4.
Res Vet Sci ; 106: 66-73, 2016 Jun.
Article in English | MEDLINE | ID: mdl-27234538

ABSTRACT

The present work aimed to assess the effect of equilibration time on post-thaw motility parameters of canine sperm frozen in three extenders: 6% low-density lipoproteins (LDL), 6% liposomes (LIPO), and 40% egg yolk plasma (EYP). A second experiment is aimed at evaluating the functional integrity of canine spermatozoa frozen in the three extenders at the best equilibration time found in the experiment one. In the first experiment, 20 ejaculates harvested from 7 dogs, were frozen in three extenders (LDL, LIPO, and EYP) after four equilibration times (30min, 1h, 3h, and 6h). The semen was evaluated after thawing using an image analyser (HT-IVOS 14.0). The 6h equilibration time gave better results of motility and progressive motility in the three studied extenders. (LDL: 58.9% vs. 42.7%; LIPO: 54.4% vs. 31.9%; EYP: 55.4% vs 40.5% for motility 6 vs. 1h). In the second experiment, 10 ejaculates taken from 6 dogs were frozen under the same conditions as the previous experiment, after 6h equilibration time. The integrity parameters of the spermatozoal membrane (hypo-osmotic swelling test, and SYBR14/propidium Iodide staining), acrosome (FITC-Pisium sativum Aglutinin staining), and DNA (acridine orange staining) were evaluated at three different stages: post-dilution (T0), post-equilibration, and post-thawing. Post-thaw results were as follows: membrane integrity (HOSt: 62;6% vs 58% vs 64.4%; SYBR14/IP: 63.6% vs 57.9% vs 64.8%); acrosome integrity (FITC-PSA: 79.4% vs 74% vs 76.2%) and DNA integrity (Acridine-orange: 98.9% vs 98.5% vs 98.7%) respectively for LDL vs. LIPO vs. EYP. No significant difference existed between the extenders tested; thus 6%LIPO and 40%EYP could be good candidates for replacement of 6%LDL in the protection of canine sperm during the freeze-thaw process without altering motility and integrity parameters.


Subject(s)
Cryopreservation/veterinary , Cryoprotective Agents/pharmacology , Dogs/physiology , Semen Preservation/veterinary , Sperm Motility/drug effects , Spermatozoa/drug effects , Acrosome/physiology , Animals , Cryopreservation/methods , Egg Yolk , Freezing , Lipoproteins, LDL , Male , Semen/drug effects , Semen Preservation/methods , Spermatozoa/physiology
5.
Theriogenology ; 84(7): 1053-60, 2015 Oct 15.
Article in English | MEDLINE | ID: mdl-26279312

ABSTRACT

The aim of this study was to develop an in vitro embryo culture medium without either fetal calf serum or BSA, using various growth factors and cytokines (GFs-CYKs; IGF-I, IGF-II, bFGF, LIF, GM-CSF, TGF-ß1, and PDGF-BB), and other molecules with surfactant and embryotrophic properties, such as recombinant albumin (RA) and hyaluronan (HA). The first part of the study was dedicated to define the best combination of GFs-CYKs + RA + HA for optimal embryonic development. Next, we compared development rates and embryo quality (inner cell mass [ICM]-to-total cell number [TCN] ratio), and postthaw survival and hatching rates using this synthetic medium (T1) and a control medium: synthetic oviduct fluid + BSA + ITS (insulin, transferrin, and selenium). The blastocyst rates were significantly higher with T1 than those with the control at 7 and 8 days after fertilization. There was no significant difference in TCN or the ICM/TCN ratio between the two treatments. Survival and hatching rates 48 hours after thawing were similar for both treatments. Finally, nine embryo transfers were conducted using fresh and previously frozen Day-7 blastocysts to evaluate the in vivo viability of embryos produced in this synthetic medium; four gestations were obtained from six fresh embryos and one gestation from three frozen embryos. In conclusion, the fetal calf serum and BSA-free medium, supplemented with GFs-CYKs + RA + HA, improved embryo development and gave comparable ICM/TCN ratios and postthaw survival rates to the control with BSA. Fresh and frozen embryos produced in this medium are viable for embryo transfer. This fully synthetic method of embryo culture is a useful means of reducing the risk of disease transmission via embryo transfer.


Subject(s)
Cattle/embryology , Culture Media , Embryo Culture Techniques/veterinary , Fertilization in Vitro/veterinary , Albumins , Animals , Blastocyst/physiology , Cryopreservation , Culture Media/chemistry , Cytokines , Embryo Culture Techniques/methods , Embryo Transfer/veterinary , Embryonic Development , Female , Hyaluronic Acid , Intercellular Signaling Peptides and Proteins , Pregnancy , Recombinant Proteins , Surface-Active Agents
6.
Mucosal Immunol ; 8(5): 1000-8, 2015 Sep.
Article in English | MEDLINE | ID: mdl-25563498

ABSTRACT

Membrane-associated mucins (MAMs) expressed on the ocular surface epithelium form a dense glycocalyx that is hypothesized to protect the cornea and conjunctiva from external insult. In this study, the hypothesis that the MAMs MUC1 and MUC16, expressed on the apical surface of the corneal epithelium, suppress Toll-like receptor (TLR)-mediated innate immune responses was tested. Using an in vitro model of corneal epithelial cells that are cultured to express MAMs, we show that reduced expression of either MUC1 or MUC16 correlates with increased message and secreted protein levels of the proinflammatory cytokines interleukin (IL)-6, IL-8, and tumor necrosis factor-α (TNF-α) following exposure of cells to the TLR2 and TLR5 agonists, heat-killed Listeria monocytogenes and flagellin, respectively. As mice express Muc1 (but not Muc16) in the corneal epithelium, a Muc1(-/-) mouse model was used to extend in vitro findings. Indeed, IL-6 and TNF-α message levels were increased in the corneal epithelium of Muc1(-/-) mice, in comparison with wild-type mice, following exposure of enucleated eyes to the TLR2 and TLR5 agonists. Our results suggest that the MAMs MUC1 and MUC16 contribute to the maintenance of immune homeostasis at the ocular surface by limiting TLR-mediated innate immune responses.


Subject(s)
CA-125 Antigen/immunology , Conjunctiva/immunology , Cornea/immunology , Immunity, Innate , Membrane Proteins/immunology , Mucin-1/immunology , Toll-Like Receptor 2/immunology , Toll-Like Receptor 5/immunology , Animals , CA-125 Antigen/genetics , Cell Line, Transformed , Conjunctiva/microbiology , Cornea/microbiology , Cytokines/genetics , Cytokines/immunology , Humans , Listeria monocytogenes/immunology , Membrane Proteins/genetics , Mice , Mice, Knockout , Mucin-1/genetics , Toll-Like Receptor 2/genetics , Toll-Like Receptor 5/genetics
7.
Prog Urol ; 23(12): 977-85, 2013 Oct.
Article in French | MEDLINE | ID: mdl-24090782

ABSTRACT

OBJECTIVE: Evaluate a self-screening questionnaire for bladder cancer of occupational origin and analyse an influence of exposure to a carcinogen bladder tumor on prognosis. PATIENTS AND METHODS: Five hundred and thirty-one patients followed, between 2005 and 2010, for bladder cancer in two university centers have received a self-screening questionnaire derived from questionnaire KVP 08. Patients who responded positively to at least one of the items were considered to have a self-screening questionnaire "positive". Patients were finally invited to take an appointment for consultation in occupational pathology. RESULTS: The response rate to self-screening questionnaire was 39.9% (212/531). It was "positive" in 82 cases (38.7%). Among the 82 patients with a self-screening questionnaire "positive", 46 patients consulted in occupational pathology (56%). Occupational exposure to a bladder carcinogen was documented in 91.3% of cases. Among the 22 patients who consulted in occupational pathology with a self-screening questionnaire "negative", an occupational exposure to a bladder carcinogen was documented in 13.6% of cases. The sensibility of the self-screening questionnaire was 91.3%, the specificity 86.4% and the accuracy 89.7%. The relative risk to have an occupational exposure if the self-screening questionnaire was "positive" was 6.69. The analysis of groups "positive" versus "negative" does not reveal any statistically significant difference in terms of tumor aggressiveness and disease-free survival. CONCLUSION: The self-screening questionnaire was considered relevant with good reliability for detection of occupational exposure to a bladder carcinogen.


Subject(s)
Diagnostic Self Evaluation , Occupational Diseases/diagnosis , Surveys and Questionnaires , Urinary Bladder Neoplasms/diagnosis , Aged , Female , Humans , Male , Retrospective Studies
8.
Prog Urol ; 22(12): 688-91, 2012 Oct.
Article in French | MEDLINE | ID: mdl-22999114

ABSTRACT

Prostatic Stromal Tumors of Uncertain Malignant Potential (STUMP) are rare tumor of the prostate of mesenchymal origin, accounting, with sarcoma for 0.1-0.2% of all malignant prostatic tumours. They however require to be individualized, to differentiate it from a benign prostatic hyperplasia or a sarcoma of the prostate. The therapeutic management should be made keeping in mind the risk of degeneration towards a malignant shape. Although the appropriate treatment is unknown, radical prostatectomy seem to be the treatment of reference, especially for young patient or for extensive lesion.


Subject(s)
Prostatic Hyperplasia/pathology , Prostatic Neoplasms/pathology , Stromal Cells/pathology , Humans , Male , Prostatectomy , Prostatic Hyperplasia/surgery , Prostatic Neoplasms/surgery , Sarcoma/pathology , Sarcoma/surgery
9.
Cancer Radiother ; 14 Suppl 1: S111-9, 2010 Nov.
Article in French | MEDLINE | ID: mdl-21129653

ABSTRACT

With 12,000 new cases each year in France, rectal cancers are a frequent entity. Concurrent fluoropyrimidin-based chemoradiation followed by a surgery including total mesorectal excision is the standard of care for locally advanced (T3-4) or node positive cancers of the mid and lower rectum. Modalities of irradiation depend on tumour location (mid versus lower rectum) and its local extension. Nevertheless, the clinical target volume (CTV) always encompasses the entire mesorectum, that goes from the peritoneal reflexion line (facing the third sacral vertebrae) to the levator ani muscles. The internal iliac lymph nodes are as well always included in the CTV. The aim of this article is to review the main epidemiological, anatomical, radiological and prognostic factors that are meaningful to define the optimal modalities of conformal radiation of rectal cancers. Definition of target volumes and organs at risk will be discussed, as well as doses and dose-constraints. A case report will be used to illustrate this article.


Subject(s)
Rectal Neoplasms/radiotherapy , Combined Modality Therapy , France/epidemiology , Humans , Lymphatic Metastasis , Prognosis , Radiotherapy, Conformal/methods , Rectal Neoplasms/drug therapy , Rectal Neoplasms/epidemiology , Rectal Neoplasms/pathology , Rectum/anatomy & histology , Rectum/radiation effects
10.
FEBS Lett ; 583(21): 3478-84, 2009 Nov 03.
Article in English | MEDLINE | ID: mdl-19818771

ABSTRACT

Our study aimed to establish the complete structure of the main dihydroxy conjugated triene issued from the lipoxygenation (soybean enzyme) of docosahexaenoic acid, named PDX, an isomer of protectin/neuroprotectin D1 (PD1/NPD1) described by Bazan and Serhan. NMR approaches and other chemical characterization (e.g. GC-MS, HPLC and LC-MS/MS) indicated that PDX is 10(S),17(S)-dihydroxy-docosahexa-4Z,7Z,11E,13Z,15E,19Z-enoic acid. The use of (18)O(2) and mass spectrometry showed that PDX is a double lipoxygenation product. Its structure differs from PD1, with E,Z,E geometry (PDX) instead of E,E,Z (PD1) and S configuration at carbon 10 instead of R. PDX inhibits human blood platelet aggregation at sub-micromolar concentrations.


Subject(s)
Docosahexaenoic Acids/chemistry , Docosahexaenoic Acids/pharmacology , Platelet Aggregation/drug effects , Arachidonate 15-Lipoxygenase/metabolism , Carbon/chemistry , Docosahexaenoic Acids/metabolism , Dose-Response Relationship, Drug , Gas Chromatography-Mass Spectrometry , Humans , Hydroxides/chemistry , Magnetic Resonance Spectroscopy , Glycine max/enzymology , Stereoisomerism
12.
Biotechnol Prog ; 19(3): 858-63, 2003.
Article in English | MEDLINE | ID: mdl-12790650

ABSTRACT

This paper presents a study about the influence of gas velocity on a methanogenic biofilm in an inverse turbulent bed reactor. Experimental results indicate a dynamic response of the growing attached biomass to the changes of hydrodynamic conditions, mainly attrition constraints. Short but intensive increases of gas velocity (U(g)) are shown to induce more detachment than a high but constant gas flow rate. Hydrodynamic conditions control the composition of the growing biofilm in terms of cells and exocellular polymeric substances (EPS). The cell fraction within the biofilm (R(cell)) was found to be inversely proportional to the gas velocity. The specific activity expressed in methane production rate or COD removal rate is higher in biofilms formed under high hydrodynamic constraints. The control of the hydrodynamic conditions in a biofilm reactor should make it possible to obtain a resistant and active biofilm.


Subject(s)
Bacteria, Anaerobic/cytology , Bacteria, Anaerobic/physiology , Biofilms/growth & development , Bioreactors/microbiology , Cell Culture Techniques/methods , Methane/metabolism , Rheology/methods , Cell Culture Techniques/instrumentation , Cell Division/physiology
13.
Toxicon ; 40(10): 1451-62, 2002 Oct.
Article in English | MEDLINE | ID: mdl-12368115

ABSTRACT

We determined the seasonal distribution of paralytic shellfish toxins (PSTs) and PST producing bacteria in > 15, 5-15, and 0.22-5 microm size fractions in the St Lawrence. We also measured PSTs in a local population of Mytilus edulis. PST concentrations were determined in each size fraction and in laboratory incubations of sub-samples by high performance liquid chromatography (HPLC), including the rigorous elimination of suspected toxin 'imposter' peaks. Mussel toxin levels were determined by mouse bioassay and HPLC. PSTs were detected in all size fractions during the summer sampling season, with 47% of the water column toxin levels associated with particles smaller than Alexandrium tamarense (< 15 microm). Even in the > 15 microm size fraction, we estimated that as much as 92% of PSTs could be associated with particles other than A. tamarense. Our results stress the importance of taking into account the potential presence of PSTs in size fractions other than that containing the known algal producer when attempting to model shellfish intoxication, especially during years of low cell abundance. Finally, our HPLC results confirmed the presence of bacteria capable of autonomous PST production in the St Lawrence as well as demonstrating their regular presence and apparent diversity in the plankton.


Subject(s)
Bacterial Toxins/metabolism , Bivalvia/chemistry , Chemical Fractionation/methods , Saxitoxin/isolation & purification , Seasons , Shellfish Poisoning , Animals , Biological Assay , Bivalvia/microbiology , Chromatography, High Pressure Liquid , Mice , Paralysis/chemically induced , Paralysis/physiopathology , Particle Size , Quebec , Saxitoxin/analogs & derivatives , Seawater , Shellfish/microbiology
14.
Invest Ophthalmol Vis Sci ; 42(11): 2483-9, 2001 Oct.
Article in English | MEDLINE | ID: mdl-11581187

ABSTRACT

PURPOSE: To determine whether the number of filled conjunctival goblet cells and mucin gene expression are altered in a mouse model of allergic conjunctivitis. METHODS: A/J mice were sensitized intraperitoneally with cat dander or the peptide P3-1 from the protein Fel d1. Two weeks later, the mice were challenged for 7 consecutive days with eye drops containing the allergens. Conjunctival tissue was harvested at 0, 6, 24, or 48 hours after final antigen challenge. Control samples were naïve animals and mice sensitized with cat dander and challenged with OVA-peptide or PBS. The mean number of filled goblet cells per square millimeter in three forniceal fields for each group was determined in wholemounts of conjunctiva prepared using rhodamine-phalloidin labeling followed by confocal microscopy. RNA was isolated from conjunctiva of the contralateral eye and taken for relative quantitation of mRNA of the goblet cell mucin Muc5AC and the epithelial membrane-spanning mucin Muc4, by real-time RT-PCR. RESULTS: The number of filled goblet cells was significantly decreased with both cat dander and P3-1, after final ocular challenge (P < 0.001). The most significant decrease over naïve mice was seen at 6 hours after final challenge with both allergens. The number of filled goblet cells was still decreased but was returning toward naïve levels at 24 hours (P < 0.05), and at 48 hours no significant difference was seen compared with naïve, PBS-treated, and OVA-peptide-treated control samples. For both cat dander and P3-1, Muc5AC and Muc4 mRNA was found to be decreased at the time of final ocular challenge. The level of Muc5AC mRNA from goblet cells rebounded from the decrease to show an increase over control by 24 hours after final challenge, and by 48 hours, the mRNA level had returned to naïve control range. In contrast, significant increases in Muc5AC mRNA were evident after final control challenge with PBS or OVA-peptide, indicating a potential irritant effect of drop application. The Muc4 mRNA level was significantly reduced at all time points except 24 hours after the last challenge. By comparison with allergen-challenged eyes, no change in Muc4 message levels was noted at any time point in OVA-peptide- or PBS-treated control eyes. CONCLUSIONS: These findings demonstrate that, in the conjunctiva of mice, repetitive application of allergens induces a reduction in the number of filled goblet cells and a decrease in Muc5AC and Muc4 mRNAs. After a period of 24 to 48 hours, the goblet cell number return to naïve levels, and goblet cell mucin mRNA levels return to above or within normal range, indicating a rapid recovery in the mucus secretion system.


Subject(s)
Conjunctiva/metabolism , Conjunctivitis, Allergic/pathology , Goblet Cells/pathology , Mucins/genetics , Allergens , Animals , Cell Count , Conjunctivitis, Allergic/metabolism , Epithelium/metabolism , Female , Glycoproteins , Mice , Mice, Inbred A , Microscopy, Confocal , Models, Animal , Mucin 5AC , Mucin-4 , Mucins/metabolism , Ovalbumin , RNA, Messenger/metabolism , Reverse Transcriptase Polymerase Chain Reaction
15.
Can J Microbiol ; 47(7): 667-9, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11547887

ABSTRACT

Rapid molecular strain typing is critical for effective outbreak investigation and implementation of infection control measures. Pulsed-field gel electrophoresis is a highly discriminatory technique for Campylobacter jejuni, but generally requires 3-5 days. We describe a simplified protocol for pulsed-field gel electrophoresis that provides high quality typing of C. jejuni isolates in a single day.


Subject(s)
Bacterial Typing Techniques , Campylobacter Infections/epidemiology , Campylobacter jejuni/classification , DNA, Bacterial/analysis , Molecular Epidemiology , Adult , Aged , Campylobacter Infections/microbiology , Campylobacter jejuni/genetics , Campylobacter jejuni/isolation & purification , Electrophoresis, Gel, Pulsed-Field , Female , Humans , Male , Middle Aged , Time Factors
16.
Am J Med Genet ; 105(4): 369-74, 2001 May 08.
Article in English | MEDLINE | ID: mdl-11378852

ABSTRACT

Estrogen and thyroid hormones exert effects on growth, development, and differentiation of the nervous system. Hormone administration can lead to changes in behavior, suggesting that genetic variants of the estrogen receptor alpha (ERalpha) and the thyroid hormone receptor alpha (TRalpha) genes may predispose to psychiatric diseases. To investigate this possibility, regions of likely functional significance (all coding exons and flanking splice junctions) of the ERalpha and TRalpha genes were scanned in patients with schizophrenia (113), along with pilot studies in patients with bipolar illness (BPI), puerperal psychosis, autism, attention-deficit hyperactivity disorder (ADHD), and alcoholism. A total of 1.18 megabases of the ERalpha gene and 1.16 megabases of the TRalpha gene were scanned with Detection of Virtually All Mutations-SSCP (DOVAM-S), a method that detects virtually all mutations. Four missense mutations, seven silent mutations and one deletion were identified in the ERalpha gene, while only four silent mutations were present in the TRalpha gene. Two of the missense mutations in ERalpha are conserved in the six available mammalian and bird species (H6Y, K299R) and a third sequence variant (P146Q) is conserved in mammals, birds, and Xenopus laevis, hinting that these sequence changes will be of functional significance. These changes were found in one patient each with BPI, puerperal psychosis, and alcoholism, respectively. Analysis of the ERalpha and TRalpha genes in 240 subjects reveals that missense changes and splice site variants are uncommon (1.7% and 0%, respectively). Further analyses are necessary to determine if the missense mutations identified in this study are associated with predisposition or outcome for either psychiatric or nonpsychiatric diseases.


Subject(s)
Receptors, Estrogen/genetics , Receptors, Thyroid Hormone/genetics , Schizophrenia/genetics , Alleles , Base Sequence , DNA/chemistry , DNA/genetics , DNA Mutational Analysis , Estrogen Receptor alpha , Gene Frequency , Humans , Mutation, Missense , Polymorphism, Genetic , Polymorphism, Single-Stranded Conformational , Schizophrenia/pathology
17.
Diabetes ; 50(3): 660-6, 2001 Mar.
Article in English | MEDLINE | ID: mdl-11246888

ABSTRACT

Atherosclerosis is a major complication of type 2 diabetes. The pathogenesis of this complication is poorly understood, but it clearly involves production in the vascular wall of macrophage (Mo) lipoprotein lipase (LPL). Mo LPL is increased in human diabetes. Peripheral factors dysregulated in diabetes, including glucose and free fatty acids (FAs), may contribute to this alteration. We previously reported that high glucose stimulates LPL production in both J774 murine and human Mo. In the present study, we evaluated the direct effect of FAs on murine Mo LPL expression and examined the involvement of peroxisome proliferator-activated receptors (PPARs) in this effect. J774 Mo were cultured for 24 h with 0.2 mmol/l unsaturated FAs (arachidonic [AA], eicosapentaenoic [EPA], and linoleic acids [LA]) and monounsaturated (oleic acid [OA]) and saturated FAs (palmitic acid [PA] and stearic acid [SA]) bound to 2% bovine serum albumin. At the end of this incubation period, Mo LPL mRNA expression, immunoreactive mass, activity, and synthetic rate were measured. Incubation of J774 cells with LA, PA, and SA significantly increased Mo LPL mRNA expression. In contrast, exposure of these cells to AA and EPA dramatically decreased this parameter. All FAs, with the exception of EPA and OA, increased extra- and intracellular LPL immunoreactive mass and activity. Intracellular LPL mass and activity paralleled extracellular LPL mass and activity in all FA-treated cells. In Mo exposed to AA, LA, and PA, an increase in Mo LPL synthetic rate was observed. To evaluate the role of PPARs in the modulatory effect of FAs on Mo LPL gene expression, DNA binding assays were performed. Results of these experiments demonstrate an enhanced binding of nuclear proteins extracted from all FA-treated Mo to the peroxisome proliferator-response element (PPRE) consensus sequence of the LPL promoter. PA-, SA-, and OA-stimulated binding activity was effectively diminished by immunoprecipitation of the nuclear proteins with anti-PPAR-alpha antibodies. In contrast, anti-PPAR-gamma antibodies only significantly decreased AA-induced binding activity. Overall, these results provide the first evidence for a direct regulatory effect of FAs on Mo LPL and suggest a potential role of PPARs in the regulation of Mo LPL gene expression by FAs.


Subject(s)
Fatty Acids/physiology , Lipoprotein Lipase/metabolism , Macrophages/enzymology , Animals , Cell Line , Consensus Sequence , Fatty Acids/pharmacology , Immunologic Techniques , Lipoprotein Lipase/genetics , Mice , Nuclear Proteins/metabolism , Promoter Regions, Genetic/genetics , Promoter Regions, Genetic/physiology , RNA, Messenger/metabolism , Receptors, Cytoplasmic and Nuclear/physiology , Response Elements/physiology , Transcription Factors/physiology
18.
J Clin Endocrinol Metab ; 86(2): 594-600, 2001 Feb.
Article in English | MEDLINE | ID: mdl-11158014

ABSTRACT

The physical character and amount of mucus secreted by the endocervix changes dramatically during the menstrual cycle to facilitate sperm migration at the time of midcycle ovulation. Mucins are highly glycosylated, high-molecular-weight proteins, which are the major structural components of the protective mucus gel covering all wet-surfaced epithelia, including that of the endocervix. We have previously demonstrated that the endocervical epithelium expresses messenger RNA (mRNA) of three of the large gel-forming mucins, designated MUC5AC, MUC5B, and MUC6, with mRNA of MUC5B predominating. Because mucin protein levels may be regulated posttranscriptionally, measurement of MUC5B protein levels with cycle are needed for correlation to mRNA levels. Measurement of specific mucin gene products within mucus secretions has been limited by availability of specific, well-characterized antibodies and by volume requirements of the isolation protocols for mucins, which include CsCl density centrifugation and fraction isolation. To measure MUC5B protein within the cervical mucus through the hormone cycle, we developed a polyclonal antibody specific to the mucin. The antibody, designated no. 799, is to a synthetic peptide mimicking a 19-amino-acid segment of an intercysteine-rich region within the D4 domain in the 3' region of the MUC5B protein. It recognizes native as well as denatured MUC5B on immunoblot, is preadsorbable with its peptide, and binds to apical secretory vesicles of epithelia expressing MUC5B. We used the MUC5B antibody along with a cervical mucin standard cervical mucin isolate in enzyme-linked immunosorbent assay to determine the relative amount of MUC5B mucin in samples of human cervical mucus taken through the menstrual cycle. We demonstrate a peak of MUC5B mucin in human cervical mucus collected at midcycle, compared with mucus from early or late in the cycle. This peak in MUC5B content coincides with the change in mucus character that occurs at midcycle, suggesting that this large mucin species may be important to sperm transit to the uterus.


Subject(s)
Cervix Mucus/physiology , Gene Expression Regulation , Menstrual Cycle/physiology , Mucins/genetics , Amino Acid Sequence , Antibody Specificity , Cervix Mucus/cytology , Enzyme-Linked Immunosorbent Assay , Epitopes/chemistry , Epitopes/immunology , Female , Humans , Luteinizing Hormone/metabolism , Molecular Sequence Data , Mucin-5B , Mucins/analysis , Mucins/blood , RNA, Messenger/analysis , Regression Analysis , Reverse Transcriptase Polymerase Chain Reaction , Saliva/chemistry
19.
J Rehabil Res Dev ; 37(1): 1-10, 2000.
Article in English | MEDLINE | ID: mdl-10847567

ABSTRACT

Differences in pelvic obliquity between small groups of persons with unilateral lower limb amputation and subjects without amputation were analyzed. Kinematic walking data were collected as six males with transtibial amputation and three males with transfemoral amputation walked over a range of speeds. The pelvic obliquity patterns and amplitudes from the groups with amputation were compared to normal data. Results showed that smaller peak-to-peak amplitudes of pelvic obliquity were associated with higher amputation levels. Pelvic drop during early prosthetic-limb stance tended to be smaller than during early sound-limb stance. Most of the subjects with amputation exhibited an obliquity pattern in which the hip on the prosthetic side was raised above the stance-side hip during prosthetic swing phase, indicative of a compensatory action known as hip-hiking. The subjects with transfemoral amputation exhibited this hip-hiking pattern during sound-limb swing phase as well. Results from this study suggest that further investigation is required to determine those limitations of current prosthetic technology that adversely affect pelvic obliquity in the gait of persons with amputation, and to determine if significant benefit can be realized by restoring a normal pattern of pelvic obliquity to the gait of persons with amputation.


Subject(s)
Amputation, Surgical/methods , Artificial Limbs , Gait/physiology , Pelvic Bones/physiology , Prosthesis Fitting , Adult , Aged , Biomechanical Phenomena , Femur/surgery , Hip Joint/physiology , Humans , Leg , Male , Middle Aged , Range of Motion, Articular , Reference Values , Tibia/surgery
20.
J Pediatr Orthop ; 20(3): 349-55, 2000.
Article in English | MEDLINE | ID: mdl-10823603

ABSTRACT

The aims of this study were to determine the influence of external tibial torsion on the effectiveness of the ankle-foot orthoses (AFO) in children with lumbosacral myelomeningocele. Forty patients with normal tibial rotation and 18 patients with excessive external tibial torsion were evaluated with three-dimensional gait analysis at their comfortable walking speed. The group with normal tibial rotation showed significantly greater knee extension and lower mean extension moment compared with the group with external tibial torsion (p < 0.05). The posteriorly and laterally deviated ground-reaction force relative to the knee-flexion axis compromises the ability of this force to facilitate knee extension. Patients with torsional magnitudes >20 degrees demand close inspection as candidates for derotation osteotomy. The AFO will continue to stabilize the ankle-foot complex, but improved knee motion, knee-extensor activity, and ultimately walking efficiency may be compromised.


Subject(s)
Gait/physiology , Hip Joint/physiopathology , Knee Joint/physiopathology , Meningomyelocele/physiopathology , Orthotic Devices , Adolescent , Biomechanical Phenomena , Child , Child, Preschool , Humans , Muscle, Skeletal/physiopathology
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