The Impact of Dose Rate on the Tumor Microenvironment Using Flattening-filter-free Beams.

AIMS: Recently, dose delivery technology has rapidly evolved with flattening filter-free beams (FFF), and the biological effects of high dose rates are a matter of interest. We hypothesized that FFF beams at different dose rates obtained with modern linear accelerators have different effects on the TME. MATERIALS AND METHODS: The B16-F10 melanoma syngeneic tumor model was established, and mice were randomized to 2 different doses (2 Gy and 10 Gy) and 3 different dose rates (1 Gy/min, 6 Gy/min, and 14 Gy/min) along with the control group. Euthanasia was performed on the seventh day after RT, and intracardiac blood was collected for a comet assay. Tumors were harvested and examined histomorphologically and immunohistochemically. Statistical analyses were performed using SPSS software version 23 (SPSS Inc., Chicago, IL, USA). RESULTS: The daily growth rate was uniform, and no difference was observed between tumor volumes across all three dose rates for each dose. Deoxyribonucleic acid (DNA) damage in blood mononuclear cells was not affected by dose or dose rate. In the TME histomorphological examination, the number of mitosis is less in the 10 Gy arm, whereas the pleomorphism score was greater. Nevertheless, varying dose rates had no effect on the number of mitosis or the pleomorphism score. The severity of the inflammation, cell densities in the TME, and expression of immunohistochemical markers were comparable across all doses and dose rates. CONCLUSION: In our study involving the B16-F10 syngeneic tumor model, varying dose rates obtained with FFF beams had no effect on tumor volume, blood mononuclear cell DNA damage, or TME parameters. However, in order to fully understand the biological impacts of novel techniques, our study should be validated with alternative preclinical setups.

Involved-site Radiotherapy Delineation Dilemmas in the Treatment of Adult Hodgkin Lymphoma: Turkish Society for Radiation Oncology Hematological Oncology, Pediatric Oncology and TBI Study Group Case-based Questionnaire Review (TROD 03-005).

AIMS: The International Lymphoma Radiation Oncology Group (ILROG) defined involved-site radiotherapy (ISRT) guidelines. These rules offer a certain variability that allows for autonomous decision-making in diverse clinical settings. However, this flexibility also gives rise to conflicts about the selection of treatment fields in the daily decision-making process. The aim of this study was to show the extent of interobserver variability when ILROG-ISRT recommendations were used in different clinical scenarios. MATERIALS AND METHODS: The 10-question survey used in our study consisted of two parts (part A and part B) and was prepared by four senior radiation oncologists experienced in the haemato-oncology field. The results were presented by stratifying according to clinical experience (<10 years, ≥10 years). Binomial tests (one-sided) were conducted to assess whether answers for each group and the whole group reached a consensus. RESULTS: Twenty-six radiation oncologists, 13 of whom had less than 10 years of experience and 13 seniors, participated in the survey. Eighty per cent of respondents thought ILROG did not bring sufficient solutions for all clinical scenarios but offered solutions in some cases. In different case-based scenarios, the consensus among the respondents decreased down to 38%. Senior radiation oncologists were found to have more doubts about the adequacy of current guidelines. CONCLUSIONS: ILROG guidelines allow for a high degree of variability in real-life clinical scenarios and different interpretation of the recommendations may lead to increased toxicity and recurrences. Therefore, there is a need for refinement in ISRT delineation strategies. On behalf of the Turkish Society for Radiation Oncology Hematological Oncology, Pediatric Oncology and TBI Study Group, we are planning to carry out further educational contouring sessions to detect the interobserver variability in real-life contouring cases.

Effects of Complement Regulators and Chemokine Receptors in Type 2 Diabetes.

CD55 and CD59 are complement regulatory proteins suggested to be related with progression of diabetes and its complications. The stromal cell-derived factor 1 (SDF-1) and C-X-C chemokine receptor type 4 (CXCR-4) are chemokine proteins. We aimed to investigate the relation of CD55 and CD59 expression levels and polymorphisms of SDF-1 and CXCR-4 with type 2 diabetes mellitus (T2DM) and its complications. Seventy-five T2DM patients and 73 controls were enrolled. Expression levels of CD55 and CD59 were measured by FACS Calibur; qRT-PCR was used to determine SDF-1 and CXCR-4 gene polymorphisms. CD55 and CD59 expressions in patients with nephropathy, retinopathy and cardiovascular disease were significantly lower than controls. Frequency of CXCR-4 T allele carrying was high in patients and created 1.6 fold risk for the disease (p = .07). CXCR-4 a allele carriers had decreased nephropathy; although there was no statistical significance in carrying CXCR-4 T allele, presence of nephropathy was approximately 2 times higher (p = .254). The nephropathy risk increased 10-fold in CXCR-4 TT genotype carriers (p = .02). All SDF-1 CC genotype carriers had retinopathy, so, it was considered that the CC genotype was effective in retinopathy development (p = .031). For the presence of cardiovascular disease, significant difference was observed for SDF-1 genotypes. Increased cardiovascular risk of 5- and 1.9-fold in SDF-1 T (p = .007) and CXCR-4 T (p = .216) allele carriers, respectively, was observed. We suggest that CD55 and CD59 protein levels and SDF-1 and CXCR-4 have predictive importance in process, complications and tendency of T2DM.

Hyaluronic acid spacer in focal prostate reirradiation: A single centre experience.

PURPOSE: The optimal management of locally recurrent prostate cancer after curative radiotherapy is still unknown. In this study, we evaluated the preliminary results of reirradiation using stereotactic body radiotherapy for locally recurrent prostate cancer after initial definitive local radiotherapy. MATERIALS AND METHODS: Between April 2016 and February 2019, 11 patients with recurrent disease at the previously irradiated prostate were treated. Local recurrence was detected by radiological with or without functional imaging modalities including prostate multiparametric/pelvic MRI or positron-emission tomography-computerised tomography with (68Ga)-labelled prostate-specific membrane antigen performed after rising prostate specific antigen serum level during follow-up. All patients received stereotactic body radiotherapy to the recurrent nodule to a total dose of 30Gy in five fractions. Hyaluronic acid spacer was injected between prostate and rectum in seven patients to decrease the rectal dose. Acute toxicity was evaluated by using Common Terminology Criteria for Adverse Events version 4.0, and late toxicity was evaluated by using Radiation Therapy Oncology Group/European Organisation for Research and Treatment of Cancer late radiation morbidity scoring schema. RESULTS: At the diagnosis, the median age was 64 years, and the mean prostate specific antigen serum concentration was 17.7ng/mL. The median interval time between local recurrence and initial definitive radiotherapy was 63 months. Mean prostate specific antigen concentration nadir value during follow-up was 0.43ng/mL. With a median follow up of 19 months, three patients developed either local or distant relapse. One patient had grade 3 acute rectal toxicity, and one patient had grade 2 late urinary toxicity. We did not observe any acute or late toxicity due to hyaluronic acid spacer injection. CONCLUSION: Reirradiation after local recurrence following initial definitive radiotherapy together with hyaluronic acid spacer use seems to be effective and safe.

An osteometric study on humerus / Estudio osteométrico del húmero

Humerus is the longest and thickest bone of the upper limb. As a long bone, it has two epiphysis and diaphysis. In this study, we aimed to conduct morphometric measurements belonging to human humerus. This study was conducted on 60 humerus (28 right, 32 left) in collections of Necmettin Erbakan University Meram Medicine Faculty Anatomy Laboratory. Digital calipers, osteometric board and precision scales for humerus bone measurements were used. Measurements were classified as measurements of diaphysis and proximal and distal epiphysis of humerus. Each bone weight was determined. Also nutrient foramen number and localization was determined. In this study, it was determined that mean right humerus length 30.41±1.73 mm, mean left humerus length 30.04±2.39 mm. It was identified that mean right humerus weight was 115.05±28.06 g, mean left humerus weigh twas 111.63±33.34 g. In 9 humerus (15 %), supratrochlear foramen has been observed. 6 of these were oval and 3 of them were round. Nutrient foramen has not been observed in two humerus (3.3 %). Also, medium and weak correlation was identified between many parameters. We believe that the obtained data from this study may be qualities of reference for sex determination from humerus.

El húmero es el hueso más largo y grueso del miembro superior. Como un hueso largo, tiene dos epífisis y una diáfisis. En este estudio, se pretende realizar mediciones morfométricas pertenecientes al húmero humano. Este estudio se realizó en 60 húmeros (28 derechos, 32 izquierdos) en colecciones del Laboratorio de Anatomía de la Facultad de Medicina Meram,Necmettin Erbakan University, Se utilizaron calibradores digitales, tableros osteométricos y escalas de precisión para medir el húmero. Las mediciones se clasificaron como medidas de húmero proximal, corporal y distal. Se determinó el peso de cada hueso. También se determinó el número y la localización de los forámenes nutricios. La longitud media del húmero derecho fue de 30,41 ± 1,73 mm y la del húmero izquierdo fue de 30,04 ± 2,39 mm. El peso medio del húmero derecho fue 115,05 ± 28,06 g y el izquierdo 111,63 ± 33,34 g. En 9 húmeros (15 %), se observó un foramen supratroclear, seis de ellos eran ovales y tres redondos.No se observó foramen nutricio en dos húmeros (3,3 %). Además, se identificó una correlación media y débil entre varios parámetros. Creemos que los datos obtenidos de este estudio pueden ser de referencia para la determinación del sexo de un individio a partir del húmero.

The synergistic effect of ω3 and Vit D3 on glycemia and TNF-α in islet transplantation.

The current treatment of type 1 diabetes consists of insulin administration. Transplantation of islets of Langerhans is considered very favorable because the full effect of insulin treatment cannot be obtained in severe cases. Although agents such as omega-3 (ω3) and vitamin D3 (Vit D3) are known to contribute to the success of islet allo-transplantation (ITX), in this study we aimed to experimentally determine their effects on glycemia and TNF-α production. Wistar albino rats, which were used as recipients, were given ω3, Vit D3, and islets by gavage, and intraperitoneal- and intraportal injections, respectively. Daclizumab (DAC) was used for immunosuppression. Glycemia levels decreased in rats treated with ω3 and vit D3. TNF-α increased in all groups due to application of STZ. After ITX (day +1), the weakest increase was observed in the ω3 + Vit D3 group. In the ITX+DAC group, compared with that of ITX only, DAC was shown to decrease levels of TNF- α following ITX, only in control group, however, similar levels of TNF-α were observed in other groups. The values in the treated groups were already lower than those of the controls in the ITX group and also remained almost equal in the ITX+DAC group. We suggest that the use of ω3 and Vit D3 together will improve the pro-inflammatory aspect encountered during and after ITXs, and contribute to the reduction of the dose of immunosuppressants in these procedures.

Effect of in situ exopolysaccharide production on physicochemical, rheological, sensory, and microstructural properties of the yogurt drink ayran: an optimization study based on fermentation kinetics.

Exopolysaccharide (EPS)-producing starter cultures are preferred for the manufacture of fermented milk products to improve rheological and technological properties. However, no clear correlation exists between EPS production and the rheological and technological properties of fermented milk products such as the yogurt drink ayran. In this study, 4 different strain conditions (EPS- and EPS+ Streptococcus thermophilus strains) were tested as a function of incubation temperature (32, 37, or 42°C) and time (2, 3, or 4 h) to determine the effect of culture type and in situ EPS production on physicochemical, rheological, sensory, and microstructural properties of ayran. Furthermore, we assessed the effect of fermentation conditions on amounts of EPS production by different EPS-producing strains during ayran production. A multifactorial design of response surface methodology was used to model linear, interaction, and quadratic effects of these variables on steady shear rheological properties of ayran samples and in situ EPS production levels. The physicochemical and microbiological characteristics of ayran samples altered depending on incubation conditions and strain selection. Steady shear tests showed that ayran samples inoculated with EPS+ strains exhibited pseudoplastic flow behavior. Production of ayran with EPS- strain (control sample) resulted in the lowest apparent viscosity values (η50), whereas those produced with the combination of 2 EPS+ strains yielded ayran with notably increased η50 values. We concluded that incubation time was the variable with the greatest effect on η50, consistency coefficient (K), and flow behavior index (n) values. In situ EPS production was also affected by these conditions during ayran fermentation in which strain-specific metabolism conditions were found to be the most important factor for EPS production. In addition, these findings correlated the amount of in situ EPS produced with the rheological properties of ayran. Scanning electron microscopy images of the samples showed differences in structural features, revealing a prominent network strand structure in the ayran samples inoculated with the admixture of 2 EPS-producing strains incubated at 37°C for 3 h. These results provide useful information for large-scale production of ayran by the dairy industry.

Multiple variations of the abdominal aorta in a single cadaver.

Numerous variations of the abdominal aorta were observed during a routine dissection of the abdominal region in a 60-year-old male cadaver in the Department of Anatomy, Meram Faculty of Medicine, Selcuk University, Turkey. In the present case, a common inferior phrenic trunk arose from the abdominal aorta and then divided into two branches. The left gastric artery arose from the front of the abdominal aorta, with an accessory right hepatic artery arising from the superior mesenteric artery. Although the single right renal artery originated from the abdominal aorta, double left renal arteries were found to originate from the abdominal aorta. Knowledge of these variations could help surgeons to identify and protect the abdominal aorta during surgery.

Intensive glucose therapy and clinical implications of recent data: a consensus statement from the Global Task Force on Glycaemic Control.

BACKGROUND: There is compelling evidence showing that achieving good glycaemic control reduces the risk of microvascular complications in people with type 1 and type 2 diabetes. Likewise, there is clear evidence to show that achieving good glycaemic control reduces the risk of macrovascular complications in type 1 diabetes. The UKPDS 10-year follow up suggests that good glycaemic control also reduces the risk of macrovascular complications in type 2 diabetes. Despite this, recent results from ACCORD, ADVANCE and VADT present conflicting results and data from the ACCORD trial appear to suggest that very low HbA(1c) targets (<6.0%) may, in fact, be dangerous in certain patient populations. AIM: To review recent results from ACCORD, ADVANCE and VADT and provide clear guidance on the clinical significance of the new data and their implications for the practising physician treating patients with type 2 diabetes. METHODS: A Pubmed search was used to identify major randomised clinical trials examining the association between glycaemic control and diabetes-associated complications. The data was reviewed and discussed by the GTF through a consensus meeting. The recommendations for clinical practice in this statement are the conclusions of these analyses and discussions. RESULTS: Evidence from ACCORD, ADVANCE, VADT and UKPDS suggests that certain patient populations, such as those with moderate diabetes duration and/or no pre-existing CVD, may benefit from intensive blood glucose control. These trials highlight the benefit of a multifactorial treatment approach to diabetes. However, ACCORD results indicate that aggressive HbA(1c) targets (<6.0%) may not be beneficial in patients with existing CVD and a longer duration of diabetes. CONCLUSIONS: Glycaemic control remains a very important component of treatment for type 2 diabetes and contrasting results from the ACCORD, ADVANCE and VADT should not discourage physicians from controlling blood glucose levels.

Growth performance and quality properties of meat from broiler chickens reared under different monochromatic light sources.

1. Quality properties of breast and drumstick muscles of female broiler chickens reared under different light sources were evaluated using a total of 480 female chicks (Ross 308). 2. After hatch, the chicks were randomly divided into three lighting treatment groups: control (daylight; mini incandescent lamps), G-B lighting group (green light monochromatic (MC) lamps for first 3 weeks switching to blue MC lamps for remaining 3 weeks), G-GB mixed lighting group (Green MC light for first 3 weeks, switching to Green MC light + Blue MC light mixture for remaining 3 weeks). 3. Feed consumption, body weight and total muscle weight values of the muscles from G-B and G-GB mix lighting groups were significantly higher than those of incandescent (control) lighting groups. 4. The breast and drumstick muscles from control lighting groups had a lower pH and water-holding capacity, but higher cooking loss values than those from G-B and G-GB mix lighting groups. The muscles from G-GB mix lighting groups exhibited the softer structure than those from other lighting groups. 5. We suggest that G-B or G-GB mix lighting during the rearing period of female broilers would improve body and muscle growth and then meat quality properties.

Effect of acute maximal exercise on lymphocyte subgroups in type 1 diabetes.

The essential therapy of diabetes mellitus includes medical nutrition therapy (MNT), exercise and medical therapy. Exercise, besides its metabolic effects, has positive influence on the immune system, but some forms of exercise may cause trauma for muscle and skeletal systems, they may also support negative effects on the immune system. Nineteen type 1 diabetic patients (mean age 22.1 +/- 2.8 yrs), followed by Diabetes Outpatient Clinic and twenty age matched male control subjects were included into the study, to demonstrate the effects of maximal, acute exercise on the immune system. The exercise test was performed according to Bruce protocol on treadmill. In diabetic subjects, increased CD19 and CD23 expressions were observed before exercise. In both groups (diabetic/control) CD3, CD4 expressions and CD4/CD8 ratio were decreased following the exercise, however expression of natural killer (NK) cells increased. Compared to type 1 diabetic patients healthy subjects had longer acute exercise that caused the increased level of CD8 expression, however type 1 diabetic patients did not show any difference. These results indicate that submaximal aerobic exercise might be recommended for type 1 diabetics without any complications because of its positive reflection on metabolic control and no negative effects on the immune system.

Characteristics of meat emulsion systems as influenced by different levels of lemon albedo.

The effect of the addition of lemon albedo on the functional properties of emulsions was studied by using a model system. Oil/water (O/W) model emulsion systems were prepared by the addition of two types of lemon albedo (raw and dehydrated) at five concentrations (0.0%, 2.5%, 5.0%, 7.5% and 10%) to mechanically deboned chicken meat. The emulsion capacity, stability, viscosity and flow properties of the prepared model emulsions were analyzed. In addition, the colour parameters of cooked emulsion gel were determined. The addition of lemon albedo increased the emulsion capacity (EC) and the highest EC value was reached with 5% of albedo added. However, further increase in the albedo concentration caused an inverse trend in the EC values. A similar trend was observed in the emulsion stability (ES) values. Dehydrated albedo (DA) addition caused higher EC and ES values than did raw albedo (RA). DA increased the L(∗), a(∗) and b(∗) values of the cooked emulsion gels. Emulsion viscosity (EV) values were positively correlated with an increase in albedo concentration and the highest EV value was obtained from the emulsions with 10% albedo. Albedo addition did not change the flow properties of the emulsions and, in addition, increased the pseudoplasticity. As a consequence, the use of lemon albedo might be a potential dietary fiber source to enhance the functional and technological properties for frankfurter-type meat products.

Intracytoplasmic cytokine levels and neutrophil functions in early clinical stage of type 1 diabetes.

Studies indicate that both CD4(+) and CD8(+) T lymphocytes and their cytokines play a critical role in different clinical stages of type 1 diabetes (T1D). Disturbances of oxidative burst and phagocytic activities in neutrophils of diabetic patients compared to uncontrolled disease support the importance of neutrophil functions in the treatment and follow up of diabetic patients. This study is designed in order to investigate Th1 and Th2 cytokine profiles and neutrophil functions in early clinical stage of T1D. Patients diagnosed as T1D but not yet under insulin therapy (Group 1; n=15) and T1D patients with disease duration of <3 months (Group 2; n=20) were compared to healthy subjects (Group 3; n=15). All subjects with T1D were positive for islet cell antibody (ICA) and glutamic acid decarboxylase antibody (GADA), their fasting glucose levels were >126 mg/dl and A1(c) levels were >8. Intracytoplasmic interleukin (IL)-2, IL-10, interferon (IFN)-gamma and tumour necrosis factor (TNF)-alpha levels of isolated CD4(+) and CD8(+) T cells, and neutrophil functions were determined by flow cytometry. Intracellular TNF-alpha level of CD4(+) T lymphocytes was significantly decreased in Group 1 compared to Group 2 and healthy subjects. In contrast, TNF-alpha in CD8(+) T lymphocytes was higher in Group 1 compared to Group 2. Increased TNF-alpha content of CD8(+) T lymphocytes was also obtained in Groups 1 and 2 compared to healthy subjects. Increased TNF-alpha secretion of CD8(+) T cells might reflect the role of CD8(+) T cells in beta cell destruction. Similar to cytokine content, phagocytic and oxidative burst activities in Group 1 were significantly lower compared to Group 2 and healthy subjects. Impaired neutrophil functions could be recovered by the treatment of the disease.

Organ specific autoantibodies in preclinical and early clinical type 1 diabetes in Turkey.

Type 1 diabetes mellitus (T1D) patients (G1; n=73) and first degree relatives with islet cell antibody (ICA) values of >or=10 JDF u twice or >or=20 JDF u one and loss of FPIR (G2; n=18) were screened for two other autoantibodies, anti-glutamic acid decarboxylase (GADA) and insulin autoantibodies (IAA), and for other organ-specific autoantibodies, anti-gastric parietal cell (anti-PCA) and anti-thyroid peroxidase (anti-TPO) as well. The two control groups consisted of healthy subjects (G3; n:55 and G4; n:13). In G1, positivity of ICA, GADA, IAA, anti-TPO and anti-PCA were 63%, 75.1%, 27.4%, 17.8% and 8.2%, respectively. In G2, positivity for GADA, IAA, anti-TPO and anti-PCA were 55.6%, 11.1%, 16.7% and 11.1%, respectively. None of the anti-TPO or anti-PCA positive cases had clinical or laboratory thyroid disease or pernicious anemia. Other organ specific antibodies, in case they accompany GADAand/or IAA in high risk individuals, result in higher risk for T1D. Moreover, this condition may indicate future potential for developing thyrogastric autoimmune diseases. In conclusion; autoantibodies are markers for autoimmune destruction in T1D, and for identification of subjects at risk for disease. Even at the time of diagnosis of T1D, screening for thyrogastric autoimmunity might be recommended for early detection of the relevant diseases.

Heart rate variability in diabetes patients.

Diabetes mellitus can cause cardiovascular autonomic neuropathy and is associated with increased cardiovascular deaths. We investigated cardiovascular autonomic neuropathy in diabetics and healthy controls by analysis of heart rate variability. Thirty-one diabetics and 30 age- and sex-matched controls were included. In the time domain we measured the mean R - R interval (NN), the standard deviation of the R - R interval index (SDNN), the standard deviation of the 5-min R - R interval mean (SDANN), the root mean square of successive R - R interval differences (RMSSD) and the percentage of beats with a consecutive R - R interval difference > 50 ms (pNN50). In the frequency domain we measured high-frequency power (HF), low-frequency power (LF) and the LF/HF ratio. Diabetes patients had lower values for time-domain and frequency-domain parameters than controls. Most heart rate variability parameters were lower in diabetes patients with chronic complications than in those without chronic complications.

Peritransplant and long-term secretion of interleukin-1beta in cyclosporine treated syngeneic rats allografted with islets of langerhans.

Interleukin-1beta (IL-1beta) is one of the proinflammatory cytokines that may mediate primary nonfunction of islets of Langerhans, limiting the success of allogeneic transplantation. The aim of this study was to assess differences between the secretion of IL-1beta as well as glycemia in peri- and long-term periods of intraportal islet allo-transplantation with or without cyclosporine (CyA) immunosuppression. Inbred Wistar albino rats were transplanted intraportally with rat islets isolated by collagenase digestion. The two recipient groups (6 rats/group) were: group 1, control, islet transplantation (ITX) without any treatment and group 2, CyA-treated via the femoral muscle on days -1, 0, +1, and +2. Serum IL-1beta (pg/mL) levels were measured by ELISA on days 0 (pre-ITX), +1, +2, and +195. Tail vein blood was used to evaluate glycemia (mg/dL). No major differences were observed in IL-1beta secretion on days 0, +1, or +195 between the groups. Immunosuppressive treatment produced significantly lower secretion in group 2 (P < .002) on day +2. Significantly greater secretions were detected at days +195, +1, and +195 compared to days 0, +2, and +2, respectively (P < .002; P < .008; P < .002). Positive correlations were observed between IL-1beta levels on days +1 and +2 (r = 0.845, P < .034). The mean values in groups 1 and 2 on days 0, +1, and +2 were 140.6 +/- 4.62 vs 119.1 +/- 12.12, 73.1 +/- 19.59 vs 88.3 +/- 14.08, 106.5 +/- 13.79 vs 92.5 +/- 15.8, respectively. No animal in group 1 displayed glycemia while three group 2 animals did at day +195. However, a negative correlation was found between IL-1beta on day 0 and glycemia on day +195 (r = -0.999, P < .026). Our results suggest that IL-1beta secretion, which is detrimental for islet engraftment, decreases at peritransplant day +2, but is upregulated during long-term graft survival both in controls and in CyA-treated recipients.

A modified automated method for isolation of viable pancreatic islets in laboratory animals.

To achieve successful islet transplantation, a high viability is required. For this reason an automated method including two chambers: one for islets isolation and one for recirculation and collection was developed. Recently, we produced a modified version of this work by building a similar system of glass where marbles were not used for disaggregation, and the pancreatic tissue had to be disrupted mechanically before the digestion phase. By using the reconfigured system, we obtained 260 +/- 20 islets from each Wistar albino rat (weighing 220 to 240 g) pancreas. Islets were observed at 35 minutes after the start of perfusion (closed circuit) and the optimum time to stop the isolation determined to be 40 minutes based upon islets viability. Although the present system is configured for islet isolation from small laboratory animals (rat, mouse), we have also obtained thousands of islets at 25 minutes after treatment of a 0.5-g piece of pig pancreas. Compared to the time-consuming manual method usually used for islet isolation from small laboratory animals, the new technique is economic, easy to use, and does not reduce islets viability.

Effect of human somatotropin hormone on cultured rat islets.

Because growth hormone (GH) improves the insulin secretion capacity of isolated human fetal islets in vitro, we sought to show that it positively influences isolated rat islets. Islets isolated from Wistar albino rats by a modified automated system were cultured in media containing 87% RPMI 1640, 10% FCS, 2% antibiotic-antimycotic, and 1% L-glutamine for 12 +/- 2 days. The cultured islets were divided into two groups: growth hormone negative (Group I) and growth hormone positive (Group II). On the 5th day we observed a decrease in the islet cell counts in both groups (Group I 28% versus Group II 45%). On the 10th day, the decrease continued in the GH-negative group (59%), while the count remained stable in the GH-positive group. The viability of rat islets was determined by fluorescein diacetate (FDA) plus propidium iodide (PI) staining. In comparison to the peripheral green, central orange-red staining pattern of Group I islets upon fluorescent microscopy, Group II showed more compact islets. Cultured islets seemed to be brighter than those without GH in the cultured islets. In conclusion, we observed that 2 weeks of incubation in the presence of GH acts positively on cultured rat islets for both their amount and their viability.

Evaluation of insulin resistant diabetes mellitus in Alström syndrome: a long-term prospective follow-up of three siblings.

Alström syndrome is a rare cause of diabetes mellitus. We studied two generations of a Turkish family in whom four members were affected by Alström syndrome. The natural course of the syndrome in three sisters was followed for 13 yr. The three sisters had short stature and truncal obesity, and developed complete blindness due to retinitis pigmentosa at 10, 5 and 13 yr of age. Two had sensorineural hearing loss and mild mental retardation, while the other developed diabetic ketoacidosis (DKA) at 14 yr and was treated with insulin from onset of diabetes. In the second case, diagnosis of diabetes was made by an OGTT at age 20 yr, and controlled with diet alone for 11 yr, then with a sulphonylurea for 2.5 yr, then with insulin. The third case developed acute hyperglycaemia at 20 yr, and required insulin from onset. Moreover, transitional features of impaired carbohydrate and fat metabolism (severe hyperinsulinaemia and insulin resistance progressing to islet beta cell failure, and hypertriglyceridaemia with fatty liver) were demonstrated, in accord with the literature. Previously unreported findings characteristic of nephro-uropathy with early-onset hypertension were also detected, and included in all cases proteinuria, glomerulopathy, and abnormal locations of the kidneys, narrowed uretero-renal junctions and dilated ureters.