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H Gerber

Publications and source records attributed to H Gerber.

At least 19 recordsLinked to original sources

Impaired insulin secretion in vivo but enhanced insulin secretion from isolated islets in pancreatic beta cell-specific vascular endothelial growth factor-A knock-out mice.

AIMS/HYPOTHESIS: Endothelial cells are considered to be essential for normal pancreatic beta cell function. However, there have been no reports showing their importance for beta cell function. MATERIALS AND METHODS: Using mice with disrupted vascular endothelial growth factor-A gene specifically in beta cells, we investigated the relation between islet vascular structure and beta cell function. RESULTS: Mice with disrupted vascular endothelial growth factor-A gene specifically in beta cells had reduced islet vascular density with impaired formation of endothelial fenestration. While their fasting glucose and body weight were comparable with control mice, their glucose- and tolbutamide-induced rapid insulin release were impaired, thus resulting in glucose intolerance. On the other hand, glucose and KCl enhanced the levels of insulin secreted from islets isolated from these mice. In addition, the production of soluble N-ethylmaleimide-sensitive factor attachment protein receptors in the islets was increased. Insulin content and expression of insulin I and pancreas duodenum homeobox 1 mRNA in the islets were also increased. CONCLUSIONS/INTERPRETATION: Our results indicate that an abnormal quality and quantity of blood vessels due to reduced expression of vascular endothelial growth factor-A in beta cells could be a cause of impaired insulin secretion without impairment of beta cell function.

Animals↗

Double-stranded secondary structures on mRNA induce type I interferon (IFN alpha/beta) production and maturation of mRNA-transfected monocyte-derived dendritic cells.

BACKGROUND: The development of dendritic cell (DC)-based vaccines using antigen-encoding mRNA requires identification of the critical parameters for efficient ex vivo loading of DCs. Exogenously delivered mRNA can induce DC activation, but the molecular mechanisms involved are unknown. The aim of the present study was to identify the means by which mRNA-dependent activation of DCs occurs. METHODS: In vitro transcribed mRNA molecules were delivered into porcine monocyte-derived DCs (MoDCs) using different non-viral gene transfer procedures. Using the green fluorescent protein (GFP) as reporter gene, as well as rhodamine-labeled RNA, intracellular delivery and transfection efficiency were assessed by confocal microscopy and flow cytometry. DC activation was monitored in terms of MHC class II and CD80/86 upregulation, as well as the production of type I interferon (IFN-alpha/beta). RESULTS: mRNA-lipofected MoDCs produced type I IFN and upregulated MHC class II and CD80/86. Computational analysis of the mRNA molecules predicted highly ordered secondary structures forming double-stranded RNA (dsRNA). This dsRNA was also detectable by immunofluorescence in mRNA-lipofected cells, using antibody specific for dsRNA. Digestion of the mRNA prior to lipofection with a double-strand-specific RNase, but not a single-strand-specific RNase, abrogated DC activation. Impairment of protein kinase R (PKR) with 2-aminopurine also interfered with the activation. CONCLUSIONS: Double-stranded secondary structures on mRNA delivered by lipofection can activate MoDCs. This could have important implications for mRNA-based immunomodulation of DCs, DC-based immunotherapy, and formulation of RNA-based vaccines. In addition, this report describes the first in vitro steps towards development of a novel large animal model system to evaluate DC-based vaccines against infectious diseases.

Animals↗

[The role of interventional pain therapy in palliative care].

The effective treatment of severe pain requires a multimodal approach with a thorough knowledge of the pharmacological properties of the individual drugs. Non-opioid analgesics are surprisingly effective and compare well to opioids. Due to the pain-induced changes in the spinal cord, the resulting tolerance and the specific effect of tumor mediators on the pain process, an effective treatment strategy requires in 10-30% of the tumor patients in the last weeks of their life an interventional approach to pain management. The common techniques involve neuroaxial delivery of local anaesthetics and opioids. There has been an abundant experience with these techniques in the perioperative setting. The indications are uncontrollable pain states and/or severe side effects of pain therapy including the cognitive deterioration with high dose opioids. The complications are mainly technical and are mostly due to catheter obstruction or disconnection. The increasing use of elastomeric pumps reduces the weight of the reservoir and allows the patient an easy adjustment of the flow rate.

Ambulatory Care↗

Pilot study of urinary iodine concentration and of biochemical thyroid parameters before and after cautious public health intervention on salt iodide content: the Swiss longitudinal 1996-2000 iodine study.

OBJECTIVE AND DESIGN: Iodide concentration (IC) in salt was cautiously increased in Switzerland (15 --> 20 ppm iodide). We evaluated the dynamics of the effect of this intervention on urinary iodine concentration (UIC, microg/l) and on thyroid parameters. SETTING: University Hospital in Bern, Switzerland. SUBJECTS: A cohort of 36 subjects (12 children, 11 women, 13 men) out of 44 were recruited. INTERVENTIONS: During the study periods PRE (May 1996-May 1998) and POST (October 1998-December 2000, subdivided into equal subperiods POST1 and POST2), that is, before and after the increase of IC in salt, subjects collected 6248 urine spots for analysis of UIC. Thyroid volumes (n=2/subject) and serum thyroid parameters (n=8/subject) were sequentially evaluated. METHODS: Average PRE-POST data were compared (multiple regression analysis). RESULTS: UIC increased overall by 5.1% (P=0.0003). Increase of UIC was highest in children (11.3%, P<0.0001), significant in women (8%, P=0.0016), but not significant in men (P=0.143). Comparison between periods POST1 and POST2 showed that UIC changed more gradually in women than in children. Thyroid volumes were normal, no nonphysiological change occurred. TSH indicated euthyroidism; it decreased in children (1.98 ==> 1.74 mU/l, P=0.04) and increased in men (1.65 ==> 1.91mU/l, P=0.025). FT3 decreased in children (P<0.004) and FT4 decreased in men (P=0.017), both within normal ranges. TSH, FT3 and FT4 were unchanged in women. FT3/FT4 ratios were stable. Anti-TPO-Ab titers were stable (P=0.9). Anti-Tg-Ab titers decreased (P=0.009). CONCLUSION: The significant UIC effects were of uncertain metabolic relevance. No pathological side effects occurred. Differential delays and penetrances of UIC increase in children and adults were hitherto unknown. The unspectacular stepwise policy seems to be safe. Our pilot results in a population with moderate iodine deficiency in women should be confirmed in population-based cluster studies. SPONSORSHIP: This work was supported by grants from the University Hospital in Bern, the Swiss Federal Office of Public Health (SFOPH), the 'Swiss National Foundation for Scientific Research' (32-49424.96), the 'Fondation Genevoise de Bienfaisance Valerie Rossi di Montelera', the 'Schweizerische Lebensversicherungs- und Rentenanstalt' and the 'Schüpbach Foundation of the University of Bern'.

Adolescent↗

[Knotting of a catheter in the plexus brachialis. A rare complication].

For the repair of an open injury to the hand, a continuous axillary block was performed in a 40- year-old male patient. Slight resistance was experienced during advancement of the catheter. Adequate surgical anaesthesia developed after the injection of 40 ml of a bupivacaine/prilocaine mixture. For postoperative analgesia an infusion of bupivacaine 0.25% was started. Three days later the removal of the catheter proved to be difficult due to a knot in the distal part of the catheter. This seems to be the first report of a knot in a catheter used for continuous axillary plexus block.

Adult↗

Urinary iodine concentration during pregnancy in an area of unstable dietary iodine intake in Switzerland.

We prospectively investigated urinary iodine concentration (UIC) in pregnant women and in female, non-pregnant controls in the canton of Berne, Switzerland, in 1992. Mean UIC of pregnant women [205 +/- 151 microg iodine/g creatinine (microg l/g Cr); no. = 153] steadily decreased from the first (236 +/- 180 microg l/g Cr; no. = 31) to the third trimester (183 +/- 111 microg l/g Cr, p < 0.0001; no. = 66) and differed significantly from that of the control group (91 +/- 37 microg l/g Cr, p < 0.0001; no. = 119). UIC increased 2.6-fold from levels indicating mild iodine deficiency in controls to the first trimester, demonstrating that high UIC during early gestation does not necessarily reflect a sufficient iodine supply to the overall population. Pregnancy is accompanied by important alterations in the regulation of thyroid function and iodine metabolism. Increased renal iodine clearance during pregnancy may explain increased UIC during early gestation, whereas increased thyroidal iodine clearance as well as the iodine shift from the maternal circulation to the growing fetal-placental unit, which both tend to lower the circulating serum levels of inorganic iodide, probably are the causes of the continuous decrease of UIC over the course of pregnancy. Mean UIC in our control group, as well as in one parallel and several consecutive investigations in the same region in the 1990s, was found to be below the actually recommended threshold, indicating a new tendency towards mild to moderate iodine deficiency. As salt is the main source of dietary iodine in Switzerland, its iodine concentration was therefore increased nationwide in 1998 for the fourth time, following increases in 1922, 1965 and 1980.

Adult↗

Swiss pilot study of individual seasonal fluctuations of urinary iodine concentration over two years: is age-dependency linked to the major source of dietary iodine?

OBJECTIVE: Because of known significant seasonal fluctuations of iodine concentration in cow's milk (ICM) in Switzerland (winter/summer (w/s) ICM ratio averages 5.6), we looked for seasonal variations and familial aspects in urinary iodine and sodium concentrations (UIC, UNaC). DESIGN: : Prospective sequential cohort investigation. SETTING: University hospital. SUBJECTS: Thirteen children (six girls, seven boys) aged 3-10 y and their parents (n=9) aged 30-47 y. INTERVENTIONS: The volunteers collected 1729 spot urine samples (5.1996-5.1998). UIC data from winter (UIC(w): December-February) and summer seasons (UIC(s): July-September) were compared with UNaC and average milk consumption. Iodine intakes from milk and salt were modelized. RESULTS: Highest seasonal fluctuations of UIC were found in six pre-school children (w/s UIC group average=1.56, significant), followed by seven school children (w/s UIC group average=1.24, N.S.); none existed in adults. UIC/UNaC showed corresponding seasonal fluctuations in children, but not in adults. Winter milk was an important iodine source for children, as proven indirectly by similar seasonal fluctuations of ICM, UIC, UIC/UNaC and an important part within UIC due to milk. CONCLUSIONS: Contribution to UIC from milk intake during winter was high in children (40-50%) and lower in adults (about 20%). Compared with children, dietary habits of adults are more complex and their iodine supply depends mainly on iodized salt and not on milk, so the effect of seasonal ICM variations on UIC is less marked. Because of significant seasonal UIC fluctuations in consumers of fresh milk products in Switzerland, results of future UIC studies conducted during summer and winter seasons should be compared cautiously, especially in young children. Furthermore, consumption of milk is to be promoted, since, besides calcium and vitamins, it is an essential source of iodine. UNaC determination should be included in epidemiologic studies if the dietary source of iodine is questioned.

Adult↗

Quantification of urinary iodine: a need for revised thresholds.

OBJECTIVE: To compare different possibilities of reporting the iodine supply in the same urine samples. Indeed, in field studies, urinary iodine concentration (I/L: micro g I/L, micro mol I/L, I/creatinine: micro g I/g creatanine, micro mol I/mol creatinine) is more readily available than excretion (I/24h micro g I/24 h, micro mol I/24h). However, confusion exists regarding the comparability of iodine supply based upon I/L, I/creatinine and I/24h, which for decades have been regarded as biochemically equivalent. DESIGN: We compared I/24h, I/L and I/creatinine in accurate 24 h collections of urine and I/L and I/creatinine in 47 spot urine samples. PATIENTS: A total of 13 subjects (Bern n=7, Brussels n=6) collected a total of 110 precise 24 h urine collections (Bern n=63, Brussels n=47). The subjects from Brussels also took a spot sample at the beginning of each 24 h collection. RESULTS: Iodine supply in both places was mildly deficient according to the criteria of WHO; all but one collection indicated an intake of >0.39 micro mol I/24h (>50 micro g I/24h). The same data presented as I/creatinine (or I/L) indicated an iodine intake of <0.39 (<50 micro g I/24h) in 5% (24%) of the samples in Bern and 23% (57%) in Brussels. Similar findings were observed for 47 spot samples. Whatever the cut-off selected, I/creatinine and I/L were systematically lower than I/24h (P<0.0002). Creatinine showed smaller CV than volume but did not perform better in defining iodine intake. CONCLUSION: Considering I/24h as a reference, both I/creatinine and I/L clearly underestimate the iodine intake in subjects with adequate proteoenergetic intake. The significant deviations observed illustrate the urgent need for establishing separate ranges for I/24h, I/creatinine and I/L. In population studies, these deviations might even be larger.

Adult↗

Further studies on the early protective responses of pigs following immunisation with high potency foot and mouth disease vaccine.

The ability of an emergency oil adjuvanted foot-and-mouth disease (FMD) vaccine to elicit early protective immunity in pigs against direct contact homologous challenge was examined. All vaccinates showed reduced viraemia and shedding of FMDV, and certain animals were protected, showing no clinical signs. IL-6, IL-8 and IL-12 were consistently detected in challenged animals that had been vaccinated. Other cytokines--IL-1, IL-2, TNF, TGF and interferons--were not detected. This demonstrates that the vaccine did not induce a systemic inflammatory response, nor a systemic elevation of T lymphocyte activity. Although the IL-6 and IL-8 did not relate to protection, IL-12 production was highest in the protected vaccinated pigs. Thus, the induction of monocytic cell activity, demonstrable by the production of IL-6, IL-8 and IL-12, appears to play a critical role in FMDV emergency vaccine induction of the innate immune defences which relate to early protection against FMD. The possible modes of defence in which such cytokine activity would be involved are discussed.

Animals↗

Differential expression of IgG Fc binding protein (FcgammaBP) in human normal thyroid tissue, thyroid adenomas and thyroid carcinomas.

The genetic events involved in thyroid carcinogenesis are still incompletely understood. Several rearrangements and mutations of oncogenes have been implicated in the development of thyroid papillary carcinomas, follicular adenomas and carcinomas. However, none of these molecular alterations is suitable either as a general marker for the diagnosis of thyroid carcinomas or to differentiate between thyroid follicular adenomas and carcinomas. In order to identify new genes with altered expression which could serve as such markers, we analyzed RNA from thyroid tumor and normal tissue using a novel technique called restriction-mediated differential display. Several differentially expressed genes were identified, including the gene for IgG Fc binding protein (FcgammaBP). Differential expression of FcgammaBP was confirmed by quantitative real-time RT-PCR. Our experiments showed that IgG Fc binding protein (FcgammaBP) is differentially expressed in normal thyroid tissue, thyroid adenomas and thyroid carcinomas. While the FcgammaBP gene is constitutively expressed in normal thyroid tissue, its expression is significantly increased in follicular thyroid adenomas and significantly decreased in papillary and follicular thyroid carcinomas. Thus, measurement of the expression levels of FcgammaBP in thyroid biopsies might help to make the otherwise difficult distinction between a thyroid follicular adenoma and a follicular carcinoma.

Adenoma↗

Porcine dendritic cells generated in vitro: morphological, phenotypic and functional properties.

Despite the central role that dendritic cells (DC) play in immune regulation and antigen presentation, little is known about porcine DC. In this study, two sources of DC were employed. Bone marrow haematopoietic cell-derived DC (BM-DC) were generated using granulocyte-macrophage colony-stimulating factor (GM-CSF) in the presence or absence of tumour necrosis factor-alpha (TNF-alpha). Monocyte-derived DC (Momicron-DC) were generated with GM-CSF and interleukin-4 (IL-4). In both systems, non-adherent cells developed with dendritic morphology, expressing high levels of major histocompatibility complex (MHC) class II. The presence of TNF-alpha increased the BM-DC yield, and enhanced T-cell stimulatory capacity. Both BM-DC and Momicron-DC expressed the pan-myeloid marker SWC3, as well as CD1 and CD80/86, but were also CD14+ and CD16+. The CD16 molecule was functional, acting as a low-affinity Fc receptor. In contrast, the CD14 on DC appeared to differ functionally from monocyte CD14: attempts to block CD14, in terms of lipopolysaccharide (LPS)-induced procoagulant activity (PCA), failed. The use of TNF-alpha or LPS for DC maturation induced up-regulation of MHC class II and/or CD80/86, but also CD14. Allogeneic mixed leucocyte reactions and staphylococcal enterotoxin B antigen presentation assays demonstrated that these DC possessed potent T-cell stimulatory capacity. No T helper cell polarization was noted. Both the BM-DC and the Momicron-DC induced a strong interferon-gamma and IL-4 response. Taken together, porcine DC generated in vitro possess certain characteristics relating them to DC from other species including humans, but the continued presence of CD14 and CD16 on mature and immature porcine DC was a notable difference.

Animals↗

Nuclear localization of epidermal growth factor and epidermal growth factor receptors in human thyroid tissues.

Epidermal growth factor (EGF) has widespread growth effects, and in some tissues proliferation is associated with the nuclear localization of EGF and epidermal growth factor receptor (EGFR). In the thyroid, EGF promotes growth but differs from thyrotropin (TSH) in inhibiting rather than stimulating functional parameters. We have therefore studied the occurrence and cellular distribution of EGF and EGFR in normal thyroid, in Graves' disease, where growth is mediated through the thyrotropin receptor (TSHR), and in a variety of human thyroid tumors. In the normal gland the staining was variable, but largely cytoplasmic, for both EGF and EGFR. In Graves' disease there was strong cytoplasmic staining for both EGF and EGFR, with frequent positive nuclei. Nuclear positivity for EGF and particularly for EGFR was also a feature of both follicular adenomas and follicular carcinomas. Interestingly, nuclear staining was almost absent in papillary carcinomas. These findings document for the first time the presence of nuclear EGF and EGFR in thyroid. Their predominant occurrence in tissues with increased growth (Graves' disease, follicular adenoma, and carcinoma) may indicate that nuclear EGF and EGFR play a role in growth regulation in these conditions. The absence of nuclear EGF and EGFR in papillary carcinomas would suggest that the role played by EGF in growth control differs between papillary carcinoma and follicular adenomas/carcinomas of the thyroid.

Adenoma↗

Porcine alveolar macrophages: poor accessory or effective suppressor cells for T-lymphocytes.

Porcine Alv-Mphi from bronchoalveolar lavages were tested for their function in an in vitro foot-and-mouth disease virus (FMDV)-specific lymphoproliferative recall response. The Alv-Mphi were seen to be poor accessory cells when compared with peripheral blood monocytes. This poor capacity was evident despite an efficient expression of SLA-DR region antigens, and other co-stimulatory adhesion molecules. It was noted that Alv-Mphi secrete relatively little interleukin 1 (IL-1beta), with or without LPS induction, even though mRNA for the cytokine could be detected. In contrast, blood monocytes with their effective accessory activity were potent secretors of IL-1. Although this IL-1beta would be important with respect to the accessory capacity of monocytic cells, it was noted that the absence of bioactive IL-1 from the Alv-Mpi cultures was not solely responsible for their poor accessory function. In fact, the Alv-Mphi produced factors which not only inhibited IL-1 bioactivity, but were also responsible for a clear suppression of lymphoproliferation. This suppressor activity was dependent on the type of monocytic cell present in the culture, being more prominent when "scavenger" phagocytes were present. Thus, the major role of Alv-Mphi s not as an accessory cell akin to monocytes, but as both a scavenger cell, related to Mphi derived from monocytes in the absence of inflammatory signals, and an immunoregulatory cell.

Animals↗

Allergen-specific IgE levels against crude mould and storage mite extracts and recombinant mould allergens in sera from horses affected with chronic bronchitis.

Immunoglobulin E antibody (IgE) levels against four recombinant (r) mould allergens (r-Aspergillus fumigatus [rAsp f] 7, 8 and 9; r-Alternaria alternata 1 [rAlta1]) and crude mould (Aspergillus fumigatus, Alternaria alternata, Penicillium notatum) and storage mite extracts were determined by ELISA in sera from 24 pulmonary sound control horses and 26 horses suffering from chronic bronchitis/bronchiolitis (CB), also called chronic obstructive pulmonary disease (COPD). Serum IgG and IgA titres were also determined against Aspergillus fumigatus extract and rAsp f 8.IgE against the crude extracts could be measured in all sera, but there was no significant difference between CB-affected and control horses. In contrast, only 8-30% of the horses, depending on the r-allergen tested, had detectable IgE levels in serum against the r-allergens. Horses with CB had significantly more often detectable IgE levels than controls against rAlt a 1 (10/26 and 3/24, respectively, p=0. 054), rAsp f 7 (13/26 and 2/24, respectively, p<0.01) and rAsp f 8 (11/26 and 1/24, respectively, p<0.01). Only four horses (three CB-affected and one healthy, p0.05) had detectable IgE levels against rAsp f 9. Furthermore, CB-affected horses were often sensitised against two or more r-allergens (13/26 of the CB-affected horses) while only one of the 24 healthy horses had positive IgE levels against more than one r-allergens. Similarly to IgE levels, no significant differences between CB-affected and healthy horses were found for IgG titres against the Aspergillus fumigatus extract. However, horses with CB had significantly higher serum IgG titres against rAsp f 8 than healthy controls (median=28 versus 10 relative ELISA units [REU], p<0.01). Additionally, horses with detectable IgE titres against rAsp f 8 had significantly higher IgG titres against this r-allergen than horses with undetectable IgE titres (median IgG titres=46 and 13 REU, respectively; p<0.01). For serum IgA titres, neither differences between healthy and CB-affected animals nor correlations between IgA and IgG or IgE titres could be found. These results show that horses suffering from CB are more often sensitised to some Aspergillus fumigatus and Alternaria alternata allergens than control horses and that they are partly sensitised to the same fungal proteins as mould-allergic human patients. Furthermore, this study shows that r-allergens allow a much more sensitive determination of specific serum antibody levels by ELISA than crude mould extracts.

Allergens↗

Hot microscopic areas of iodine-deficient euthyroid goitres contain constitutively activating TSH receptor mutations.

Constitutively activating TSH receptor mutations have been established as the most common molecular basis for the pathogenesis of toxic thyroid nodules. These mutations result in uncontrolled signalling through the TSH receptor that is likely to cause hyperfunction and proliferation. The incidence of toxic multinodular goitres has been demonstrated to be related to iodine deficiency. Moreover, scintigraphically autonomous areas are found in 40% of euthyroid goitres from iodine-deficient areas. To investigate the molecular cause of these autonomous areas, small autoradiographically hot areas were examined for somatic TSH receptor mutations using archival tissue sections from 14 patients with euthyroid goitres, which had been originally prepared nearly 20 years ago. All patients had received (125)I 17 h preoperatively for the autoradiographic investigation of their thyroid. Areas with high and low (125)I-labelling on autoradiography sections were collected separately either from serial paraffin-embedded tissue sections, or Eukitt-embedded tissue sections containing the autoradiograph. After genomic DNA extraction, the transmembrane segment of the TSH receptor was PCR-amplified and directly sequenced. Somatic TSH receptor mutations were identified in areas with high (125)I-labelling in four patients: A623I, L629P, F631L, and T632I. This is the first evidence that TSH receptor mutations occur in microscopic areas with increased (125)I-labelling in euthyroid goiters and it suggests that TSH receptor mutations in these areas confer the potential to develop into toxic thyroid nodules. It is therefore very likely that toxic thyroid nodules originate from small autonomous areas in iodine-deficient euthyroid goitres that contain a TSH receptor mutation.

Female↗

Functional outcome of transplanted knee joints in dogs.

Total knee joint transplantation has been performed in animal models and humans. This study investigates the impact of this operation on knee joint function in a dog model. Therefore, replantation was compared to transplantation during a 6-month follow-up period in four dogs in each group. The peak vertical ground reaction force normalized in all legs undergoing replantation and in two of four after transplantation. A third transplant recipient reduced loading from the 4th month due to a local complication, and the fourth succumbed to sepsis 3 months postoperatively. A weight-bearing index (WBI), defined as loading of the grafted divided by loading of both hind-limbs decreased from 0.48 +/- 0.08 preoperatively to 0.13 +/- 0.10 by 1 month after replantation and from 0.53 +/- 0.07 to 0 after transplantation. After 6 months, weight-bearing of all replant recipients was restored, but reduced in two transplant recipients with graft function. Full recovery after replantation, but impaired function after transplantation, was also reflected in the histological results: normal histological pictures of blood vessels, cartilage, bone and soft tissues were found in all replant recipients, but infiltrative vasculopathy indicating chronic rejection was found in the transplanted joints. The results of this animal study confirm that the procedure can lead to satisfactory functional results but also emphasize the need for perfect control of immunosuppression.

Animals↗