Production and Characterization of a Conditionally Immortalized Dog Beta-Cell Line from Fetal Canine Pancreas.

Since the 1970s, rodent and human insulin-secreting pancreatic beta-cell lines have been developed and found useful for studying beta-cell biology. Surprisingly, although the dog has been widely used as a translational model for diabetes, no canine insulin-secreting beta cells have ever been produced. Here, a targeted oncogenesis protocol previously described by some of us for generating human beta cells was adapted to produce canine beta cells. Canine fetal pancreata were obtained by cesarean section between 42 and 55 days of gestation, and fragments of fetal glands were transduced with a lentiviral vector expressing SV40LT under the control of the insulin promoter. Two Lox P sites flanking the sequence allowed subsequent transgene excision by Cre recombinase expression. When grafted into SCID mice, these transduced pancreata formed insulinomas. ACT-164 is the cell line described in this report. Insulin mRNA expression and protein content were lower than reported with adult cells, but the ACT-164 cells were functional, and their insulin production in vitro increased under glucose stimulation. Transgene excision upon Cre expression arrested proliferation and enhanced insulin expression and production. When grafted in SCID mice, intact and excised cells reversed chemically induced diabetes. We have thus produced an excisable canine beta-cell line. These cells may play an important role in the study of several aspects of the cell transplantation procedure including the encapsulation process, which is difficult to investigate in rodents. Although much more work is needed to improve the excision procedure and achieve 100% removal of large T antigen expression, we have shown that functional cells can be obtained and might in the future be used for replacement therapy in diabetic dogs.

Production, Characterization, and Function of Pseudoislets from Perinatal Canine Pancreas.

We evaluated the cell composition and function of canine pancreatic pseudoislets (PIs) produced from 42- to 55-day-old fetuses, 1- to 21-day-old pups, and an adult dog pancreas. After mild collagenase treatment, partially digested tissues were cultured for 2-3 weeks. PI production started on culture day 3, was marked for 6 to 9 days, and then stopped. PI production was greatest with the neonatal specimens, reaching about 12 million aggregates per litter (55-day-old fetus) or per pancreas (1-day-old pup). Cell composition at all stages was similar to that in adult pancreatic islets, with predominant ß cells, scant α cells and, most importantly, presence of δ cells. Among pancreatic markers assessed by quantitative real-time PCR (qRT-PCR) mRNA assay, insulin showed the highest expression levels in PIs from newborn and adult pancreas, although these were more than 1000 times lower than in adult islets. Pdx1 mRNA expression was high in PIs from 55-day-old pancreases and was lower at later stages. Consistent with the qRT-PCR results, the insulin content was far lower than reported in adult dog pancreatic islets. However, insulin release by PIs from 1-day-old pups was demonstrated and was stimulated by a high-glucose medium. PIs were transplanted into euglycemic and diabetic SCID mice. In euglycemic animals, the transplant cell composition underwent maturation and transplants were still viable after 6 months. In diabetic mice, the PI transplants produced insulin and partially controlled the hyperglycemia. These data indicate that PIs can be produced ex vivo from canine fetal or postnatal pancreases. Although functional PIs can be obtained, the production yield is most likely insufficient to meet the requirements for diabetic dog transplantation without further innovation in cell culture amplification.

Reassessment of antibiotic therapy in hospitals.

INTRODUCTION: French national guidelines state that antibiotic therapies should be reassessed between 48 and 72hours after treatment initiation and that reassessment of antibiotic therapy (RA) must be recorded in patients' files. OBJECTIVE: To determine whether RA is performed and recorded in patients' files in hospitals in a region of France. METHODS: Setting: hospitals participating in the National nosocomial infection point- prevalence survey (NPS) in Upper-Normandy, France. Patients included those receiving antibiotic therapy (excluding antibiotic prophylaxis) on NPS day, started in the hospital in which the survey was conducted and ongoing for more than 72hours. Data collected included characteristics of participating hospitals and, for each included patient, characteristics of ward, infection and antibiotic therapy, and mention in the patients' files of explicit or implicit RA. The rate of explicit and implicit RA was calculated and factors associated with explicit or implicit RA were evaluated using a univariate analysis. RESULTS: Thirty-three hospitals representing 87% of hospital beds region-wide were included in the study. In addition, 933 prescriptions were assessed for 724 infections in 676 patients. The overall rate of RA was 67.6% (49.3% of explicit RA and 18.3% of implicit RA). The rate of RA differed significantly according to infection and antibiotic class but not according to hospital or ward characteristics. CONCLUSION: Our study provides new and reassuring results regarding reassessment of antibiotic therapy.

The risk for hepatitis C infection in blood donors in Cluj County, Romania.

Blood products safety is based on different criteria including the selection of blood donors. Blood donors referred to Cluj County (Romania) Blood Transfusion Centre in January-March 2012 completed a self-administered questionnaire and were examined by a physician. Data collected from first-time and repeat donors were compared for possible risk factors for hepatitis C infection. In total, 1100 donors were selected. In first-time donors, most frequent factors were age<26 years, female gender and history of health care procedures. Behavioural risk factors (e.g. drug use, sexual promiscuity) may not be properly filtered out in blood donors, suggesting the necessity of improving the health screening process.

The seroprevalence of hepatitis C virus infection among blood donors from the North West region of Romania.

The infection with hepatitis C virus (HCV) known also as a blood-borne infection can be life-threatening by delayed consequences of persistent infection. This study aimed to estimate the hepatitis C infection prevalence, its epidemiological mode of manifestation and its recent evolution among blood donors from North West region of Romania. We made a chronological study using the HCV seropositive confirmed results to serological blood donors screening, provided by two Blood Transfusion Centres, chosen for their quality of provided data: Cluj and Satu Mare. The statistics were descriptive and the time series modelling used the polynomial regression. In both counties, about three fourth of donors were male and in Satu Mare, female donors (29.2%; p < 0.001) have been significantly more represented than in Cluj (24.6%). Hepatitis C virus seroprevalence was significantly (p < 0.01) higher in Satu Mare (279.8/10(5) donors, CI: 241.7-317.9/10(5) donors) than in Cluj (212.6/10(5) donors, CI: 187.4-237.8/10(5) donors). The mode of manifestation was endemic in different patterns, one with wave and a stable one with a particular high value of 1.091.1/10(5) female donors (CI: 674-1508.2/10(5) donors) seroprevalence, in 2008, in Satu Mare. The variation of the prevalence value was higher for women and particularly (coefficient of variation: 91.9%) for those from Satu Mare. The hepatitis C virus seroprevalence among blood donors reflected a stable, endemic manifestation with particular variation and high transmission in female populational subgroups.

Evolution of IGF-1 in children born small for gestational age and with growth retardation, treated by growth hormone adapted to IGF-1 levels after 1 year.

AIM: This study was designed to estimate the percentage of growth hormone (GH)-treated children born small for gestational age (SGA), with serum IGF-1 >2 SDS before and after GH dose adaptation. METHODS: SGA boys aged 4-9 and girls aged 4-7 with a height <-2 SDS and an annual growth rate below the mean received a subcutaneous GH dose of 57 µg/kg/day for 2 years. The GH dose was to be decreased by 30% in children with serum IGF-1 >2 SDS at 12 months and on the previous sample. The GH dose could be reduced a second time to 35 µg/kg·day. IGF-1 and IGFBP-3 dosages were centralized. RESULTS: Among the 49 (21 boys) children included in the study, 8 (16.3%) had an IGF-1 >2 SDS consecutively at 9 and 12 months (95% CI 7.3, 29.7). The GH dose was decreased in 6/8 children. However, IGF-1 levels were elevated at several nonconsecutive determinations in 45% (95% CI 28.4, 56.6) of the patients. CONCLUSION: A high IGF-1 level is observed in 45% of the GH SGA-treated children with a relatively high dose of GH. A 30% reduction in the GH dose causes a decrease in IGF-1 below 2 SDS in most children.

[Obstacles to effective treatment of depression. A general practitioners' postal survey in the north-west region of France]. / Enquête sur la prise en charge des patients dépressifs en soins primaires: les médecins généralistes ont des difficultés et des solutions.

CONTEXT AND AIM: Depression is a quite common condition, and its treatment is mainly provided by General Practitioner (GP). It is already known that detection and treatment requires significant improvement. The well known and high consumption of antidepressant drugs in France, the highest of all other European countries, requires specific studies. The causes of this situation are not clear and seem to be numerous: Patient's demands, social claims; lack of initial and continuous medical education, bad GP demographic trends, and lack of them in rural areas; pharmaceutical company pressure; and organisation of the health care system. GP are the main medical actors of the primary care system in France. The aim of this study was to survey GP perceptions on secondary care services, seek the views and barriers to the provision of good services, and ask them about perceptions and solutions they could suggest. METHODS: A structured postal questionnaire was sent to all GP of the north-west region of France, asking physicians about obstacles perceived when taking care of depressive patients; factors influencing the use of services, specialised advice, treatments, access to psychiatrists and psychological care. Their psychiatric knowledge and demographic data were also assessed. Quantitative data were analysed using Epi-Info software, and qualitative data were transcribed and coded manually. RESULTS: A total of 25% of the GP returned the questionnaire (n=2097 in 8709). The sample profile was the same as the studied population. Less than a third of the GP (28%) were aware of the clinical guidelines on depression, and less than a fifth (18%) had clinical experience of psychiatry during their studies. Lack of time was not the main obstacle assessed by the GP. Their complaints were about lack of mental health services, difficulty in accessing services, and about general liaison between primary and secondary health care services: they reported difficulties obtaining quick and good response from the specialist either for emergency or non emergency cases. Regarding secondary care, they mainly referred to the psychiatrist, rather than to the psychologist, probably because this second option is not reimbursed by the social security system. Not surprisingly, medication was cited as the most frequently used treatment, followed by psychotherapy and cognitive behavioral therapy (CBT), and almost never self help literature and self help groups. Trained GP considered they were much more comfortable coping with depressed patients, less frequently using secondary care providers, and easily alternative solutions rather than antidepressant drugs. This situation suggests the usefulness of medical education, and is attested by many qualitative answers. DISCUSSION: It is not sure that the low rate of knowledge of the guidelines should be judged only as a lack of professionalism. According to the "French Society of Primary Care", clinical guidelines need updating, and it is known that those available could be useful only for half of the situations encountered in primary care. Operational propositions urgently need to be proposed. Recent questioning of the real interest of pharmaceutical options in the treatment of depression is another argument. Nor can we wait for a hypothetic rise in the demographic situation. The GP have several propositions to improve these problems, e.g. continuous medical education (CME) focusing on "patient centred therapy", dedicated hotline or circuit for depressed people, and an adapted sociomedical directory. They also feel that political awareness about lack of physicians is required, but say that improving quality of care does not rely only on improving demographics. They ask for funds for psychological care. When thinking about the circuit of care, the role of all care providers, and their communication, a global vision appears unavoidable, which would get rid of the divisions between out-patients and the hospital. CONCLUSION: Despite an unavoidable questioning on the dysfunctions of the health care system, quality of care and probably pharmaceutical consumption for the depressed patient might be improved by simple tools, such as adapted CME for primary care physicians, and communication improvement between secondary and primary care systems.

[Information for patients about hospital infections in psychiatry: An assessment of healthcare professionals' knowledge, opinion and attitude]. / Connaissances, opinions et pratiques des professionnels concernant l'information des patients sur les infections nosocomiales : évaluation en milieu psychiatrique.

INTRODUCTION: French legislation makes mandatory for healthcare providers the disclosure of hospital infection (HI) risk and actual occurrence to the patient. Given the specific diseases encountered in psychiatry, some difficulties may be expected in practical application of this regulation. OBJECTIVES: The aim of our study was to describe the knowledge, declared practices and opinions of healthcare workers (HCW) in psychiatry concerning information for patients about HI. METHODS: We randomly selected doctors, nurses and head nurses from four hospitals with psychiatric activity in Normandy. The HCW were asked to self-complete an anonymous questionnaire, including data describing the responding HCW and questions aiming at describing his/her knowledge, attitude in routine daily practice and opinion about information to patients about HI. RESULTS: One hundred and forty-one HCW were initially selected, of which 114 (80.9%) eventually agreed to complete the questionnaire. Only eight HCW (7.0%) were considered to have a correct overall knowledge of legal obligations. Main errors concerned the obligation to inform the patient of the HI risk according to the medical procedures that are to be performed (43.9% of correct answers) and the obligation to inform the patient of the HI risk according to his/her medical condition (46.5%). The obligation to inform the patient of the occurrence of a HI was largely known (84.2%). HCW usually giving information about the risk of HI to patients without HI accounted for 5.3%. Main reasons advocated for not informing patients were a low level risk of HI in psychiatry (80.4%) and the lack of patients' demand (59.8%). In the case of HI occurrence, the percentage of HCW routinely informing patients was 13.2%. HCW systematically informing the patient's family about the occurrence of HI accounted for 9.6%. A large proportion of HCW supported delivering information to patients about HI (86.0%). HCW expected from information better approval of prevention programs by the patients (87.7%) but feared an increased anxiety in patients (75.4%) and a higher rate of care refusal (48.2%). CONCLUSION: Whereas a very large proportion of HCW in psychiatry support delivering information to patients about HI, our study shows HCW's lack of awareness of regulations and lack of declared practices. Among factors explaining this contrast, a lower perceived HI risk and severity level are to be mentioned. Training programs focusing on risk and mechanisms of HI could be offered to professionals in psychiatry. The issue of specific communication difficulties with psychiatric patients should be addressed as well. In order to develop information on HI, specific methods suited to those patients should be developed.

Influence of infection control report cards on patients' choice of hospital: pilot survey.

The impact on patients' attitudes of quality report cards on infection control in hospitals has never previously been studied. In 2006, the French government implemented a mandatory report card on infection control activity (ICALIN) in all hospitals. This approach was aimed at encouraging professionals to change their routine practices in case they should lose patients due to a low ICALIN score. Our objective was to assess what impact ICALIN could have on patients' attitude as regards hospital choice. We performed a survey of patients and visitors in 14 randomly selected hospitals of various ICALIN scores. A convenience sample of 381 patients and visitors completed an anonymous questionnaire on ICALIN, their reasons for choosing a hospital and attitude in the event of a low ICALIN score. Factors associated with interest in ICALIN and impact of ICALIN on hospital choice were assessed by logistic regression. Our results showed that 77% of participants were interested in ICALIN. ICALIN was ranked sixth as a reason for choosing a hospital. In the case of a low ICALIN, 24.1% of participants would refuse admission and 54.9% would seek advice from their general practitioner. Sociodemographic factors had no influence on patients' attitude. In conclusion, our survey suggests that patients take note of poor performance on infection control report cards. As most patients rely on their general practitioner to interpret these report cards, there is a definite need for further communication with general practitioners on this issue.

Mycoplasma hominis wound infection after a vascular allograft.

Cases of Mycoplasma hominis infections after allograft are rare. We report a case of M. hominis wound infection after a vascular allograft. The allograft was positive before having any contact with the recipient, and our investigation suggests that M. hominis may have been transmitted from the donor to the recipient. It is not clear, however, whether specific diagnosis of M. hominis should be performed on tissue before grafting in order to prevent such donor-to-host transmission.

What factors influence healthcare professionals' opinion and attitude regarding information for patients about hospital infection?

Although informing patients about medical risks is said to decrease the number of malpractice claims, most inpatients receive no information about hospital infection. Using a self-administered questionnaire, we surveyed 1270 healthcare workers randomly selected from 22 French hospitals to assess their opinion on information for patients about hospital infection risks, and their practice of informing patients with, or without, hospital infection. The influence of healthcare worker characteristics on opinion and practice was assessed using logistic regression. Response rate was 87.2%. Although 85.4% supported giving more information, only 17.0% routinely informed non-infected patients and 31.6% informed infected patients about infection. Attitudes were influenced by healthcare worker characteristics and environmental factors. Knowledge of obligations influenced practice when informing non-infected patients, but not those with hospital-acquired infection. Further research is needed to help healthcare professionals improve risk communication and disclosure of hospital infection.