Role of adipose tissue as an endocrineorgan in systemic inflammation

Introduction The escalating public health problem represented by obesity has spurred multidisciplinary research into adipose tissue and importantly, the molecular biology of the adipocyte. The concept of adipose tissue as an endocrine organ, in addition to an energy storage compartment, is now pivotal in linking excess adiposity to disease states. Recent studies suggest that obesity related metabolic disorders are characterised by mild chronic inflammation as a result of adipocytokine production from fat tissue leading to dysregulation in the pro/anti‐inflammatory systemic balance. Adipokines and pro‐inflammatory markers are implicated in insulin insensitivity, blood glucose dysregulation, inflammation and atherosclerosis. There is a considerable amount of research into the characterization of adipokines and pro‐inflammatory cytokines, the antiinflammatory adipocytokines warrant further exploration. Research studies and design This PhD research was set out to investigate potential antiinflammatory molecules that could be used as markers of, and therapeutics for, metabolic syndrome associated maladies. This PhD consists of three studies including Study 1: a haracterisation study of pro/anti‐inflammatory mediators carried out on 116 men of various BMI and body composition; Study 2: an in vitro study design carried out in human Simpson Golabi Behmel Syndrome (SGBS) adipocyte cell line investigating a glucocorticoid regulated anti‐inflammatory protein, annexin A1 (AnxA1) and its role in fat tissue function; and Study 3: a double‐blind cross‐over randomised trial in 15 borderline metabolic syndrome males investigating the effect of a supplemental antiinflammatory agent, resveratrol (250 mg/day for two weeks, from 500 mg of Polygonum cuspidatum (from root)), on metabolic parameters. Key findings Study 1: We demonstrated for the first time that AnxA1 is significantly inversely correlated with increasing BMI (R = ‐0.424**, P < 0.001), increasing body fat % (R = ‐0.192, P = 0.037) and a larger waist size (R = ‐0.390**, P < 0.001) in 118 men aged 19 to 61 years, with BMI between 16.8 – 56.4 kg/m2, BF % between 4.3 to 51.8 %. The negative correlation of decreasing plasma AnxA1 was strongest statistically when compared with WHR, rather than total body fat, suggesting that centrally located fat may be more influential at reducing plasma AnxA1 concentrations. Study 2: We have shown that ANXA1 gene is expressed in human SGBS adipocytes and hypoxia reduces the expression of ANXA1 gene showing that AnxA1 may act as a counter regulator of adipose tissue inflammation. We found that CRP expression was significantly down‐regulated following 4 (P=0.015), 8 (P=0.035) and 24 (P=0.037) of hypoxia treatment in the cells also treated with Ac2‐26 peptide compared to vehicle alone. IL‐6 was also found to be significantly down‐regulated after 24 hour hypoxia treatment in the Ac2‐26 treated cells compared to vehicle (P=0.022). Study 3: The effect of resveratrol on metabolic function had no significant effect on the metabolic markers measured including blood pressure, blood glucose, blood cholesterol and glycated LDL. Conclusion: These data demonstrate that AnxA1 could potentially represent a (fat) depot specific biomarker whose decline with increasing central adiposity may relate to the phenomena of increasing systemic inflammation and associated disease risk. We also demonstrate for the first time that an AnxA1 is expressed in human SGBS preadipocytes and mature adipocytes and AnxA1 mimetic, Ac2‐26 peptide, regulates pro‐inflammatory markers in human SGBS adipocytes. We showed that it may be difficult to improve the metabolic profile of individuals through supplementation of exogenous anti‐inflammatory agent, resveratrol. Whilst anti‐inflammatory agents such as AnxA1 may propose novel therapeutics for metabolic syndrome associated diseases, to date regular exercise and weight loss remain the main interventions that significantly cut the risk of developing chronic long‐term conditions and obesity‐associated maladies

Monacha samsunensis (Pfeiffer, 1868): another Anatolian species introduced to Western Europe, where it is known as Monacha atacis Gittenberger & de Winter, 1985 (Gastropoda: Eupulmonata: Hygromiidae)

Populations of Monacha atacis from southern Occitania in France and of M. samsunensis from northern Anatolia in Turkey (Atakum/Samsun and Kastamonu) were investigated by an integrative approach based on morphological (shell and genitalia) and molecular (mitochondrial and nuclear gene sequences) features. Morphological examination revealed a complex pattern of variation within and between geographically separated populations, while molecular analysis showed strong similarity between the two species, confirming earlier suggestions that the species are conspecific. Pfeiffer’s name Helix samsunensis introduced in 1868 has priority over the name M. atacis given by Gittenberger &amp; de Winter in 1985. © 2022 The Author(s). Published by Informa UK Limited, trading as Taylor &amp; Francis Group

Effectiveness of „mobile” and stationary X-ray units and computed tomography in brachytherapy treatment planning

CT, mobile and stationary x-ray cameras were used with the aim of comparing the source localization effectiveness in brachytherapy planning. Properties of orthogonal X-ray pictures were discussed and their impact on dose planning in brachytherapy was evaluated.Differences between doses calculated for applicator positions localized by stationary and „mobile” X-ray units ranged between 6% and 11% in the rectum and 10% in the bladder, respectively

Intravaginal brachytherapy for patients with endometrial cancer after surgery-review of technical developments

The use of intravaginal brachytherapy as a post-operative procedure to reduce the incidence of reccurence of carcinoma of the endometrium is well known. We analysed 3 differents methods of intravaginal brachytherapy: conventional brachytherapy Ra-226, LDR after-loading technic Cez 137 and HDR after-loading brachytherapy lrydium 192. In the period 1953–1986 in Gynaecological Radiotherapy Department in Poznań, brachytherapy with vaginal applicators containing 30 mg radium, filtrated by 2 mm Pb, were used after total hysterectomy. The given dose was 3000 mgh in 100 hours of one insertion. Since 1986 Caesium 137 in one oblong applicator has been used to fill the vagina. Usualy four sources were employed and treatment time was about 24 hours. On the basis of the radiological verification in two planes, the doses were calculated at 0,5 cm from the applicator surface and at the contact point of the contrast image of the Foley catheter placed in the bladder neck. Dose in the rectum was calculated at the distance shown by a marker situated in the rectum. The patients were treated to the total dose of 30 Gy. From 1995 HDR after-loading inreasingly replaced LDR after-loading in intravaginal brachytherapy. With iridium 192 the overall dose was applied in three fractions-each 6 Gy calculated at 0,5 cm from the surface of the oblong applicator. Complications were graded with EORTC\RTOG criteria

28. Comparison of doses measured by thermolumi-nescent and semiconductor detectors during total body irradiation at Cobalt-60 and 15 meV linear accelerator

In-vivo dosimetry is an important way of dose verification during total body irradiation (TBI).AimThe aim of this paper was to compare the doses measured in-vivo with two types of detectors: thermoluminescent (TLD) and semiconductor (SEM) during TBI.PatientsSince 1993, 38 patients have TBI performed, out of them – 22 on Cobalt-60 and 16 on 15 MeV linear accelerator. Total dose of 12,6 Gy was prescribed and delivered in 8 fractions during 4 days. Combination of lateral and anterior-posterior fields, with lung shields was used. Doses were measured with the aim to verify primarily calculated doses (in ten reference points in the body).MethodsMeasured doses were normalised to those pre-calculated. Mean doses and their standard deviations (SD) were calculated separately for each of ten sections, for doses measured with TLD and SEM detectors respectively. Analysis was carried out for doses measured in points lying on the beam to the body entry during irradiation at lateral fields.ResultsMean dose for the whole group of patients treated on Cobalt-60, for all ten sections together, measured with TLD detectors was equal to 1,05 (normalised to calculated dose) with standard deviation (SD) of 3,4% and for SEM was equal to 0,98 with SD = 2,5%. Respectively, for 15 MeV linear accelerator mean dose for TLD was 1,05 with SD = 3,1% and 1,02 with SD = 3,1%.ConclusionsMean differences between doses measured with TLD and SEM dosimeters at beam entry at lateral fields were equal to 6,8% for Cobalt-60 and 3,1% for 15 MeV

Effect of irradiation on interleukin 6 and soluble interleukin 6 receptor modified melanoma genetic vaccine

We have designed phase I/II human melanoma gene therapy clinical protocol. The aim of the study was to actively immunize HLA-A1 and/or HLA-A2-positive patients with melanoma with an admixture of irradiated autologous tumor cells and allogeneic melanoma cells genetically engineered to secrete IL-6 and sIL-6R in order to elicit or enhance specific and nonspecific antimelanoma immune responses to autologous tumor cells to eradicate distant melanoma lesions. Irradiation of autologous and allogeneic tumor cells is a key step in preparation of cellular vaccine because of two major reasons, (i) it inhibits cell proliferation which is crucial in the case of autologous cells which may form a tumor; (ii) it increases melanoma vaccine immunogenicity. The aim of the study was to estimate the optimal dose of ionizing radiation which will provide sterilization of both autologous and allogeneic melanoma cells and will ensure cytokine secretion.Human melanoma cells (Mich-1) were transduced with IL-6 and sIL-6R cDNA using double copy bicistronic retroviral vector. Parental and transduced cells were seeded at in six-well tissue culture plates and were irradiated with 10, 50, 100 and 200 Gy. Secretion of both recombinant proteins into culture was analyzed before and 24, 48,72,96 h and 6, 7, 10 and 12 days following irradiation. At the same time adherent cells were enumerated, evaluated’ for viability and proliferation. At 24, 48, 72 and 96 h postirradiation specific IL-6 and sIL-6R mRNA levels were analyzed.Irradiation of gene modified cells inhibited their proliferation in the dose dependant manner. Dose of 50 Gy sufficiently affected cell proliferation, however, for safety reasons we decided to use the dose of 100 Gy for vaccine preparation. Irradiation did not inhibit secretion of IL-6 and sIL-6R. In contrary, on a per cell basis it significantly increased their secretion which lasted 12 days postirradiation. Interestingly, we did not observe dose or time dependent differences in specific mRNA cellular levels suggesting that increased secretion of both proteins is regulated not on the transcriptional but rather on the posttranscriptional level. Taking all these facts into account we concluded that irradiation of tumor cells may provide an effective and safe approach for gene-modified vaccine preparation

Effect of irradiation on interleukin 6 and soluble interleukin 6 receptor modified melanoma genetic vaccine

We have designed phase I/II human melanoma gene therapy clinical protocol. The aim of the study was to actively immunize HLA-A1 and/or HLA-A2-positive patients with melanoma with an admixture of irradiated autologous tumor cells and allogeneic melanoma cells genetically engineered to secrete IL-6 and sIL-6R in order to elicit or enhance specific and nonspecific anti-melanoma immune responses to autologous tumor cells to eradicate distant melanoma lesions. Irradiation of autologous and allogeneic tumor cells is a key step in preparation of cellular vaccine because of two major reasons, (i) it inhibits cell proliferation which is crucial in the case of autologous cells which may form a tumor; (ii) it increases melanoma vaccine immunogenicity. The aim of the study was to estimate the optimal dose of ionizing radiation which will provide sterilization of both autologous and allogeneic melanoma cells and will ensure cytokine secretion.Human melanoma cells (Mich-1) were transduced with IL-6 and sIL-6R cDNA using double copy bicistronic retroviral vector. Parental and transduced cells were seeded at in six-well tissue culture plates and were irradiated with 10, 50, 100 and 200 Gy. Secretion of both recombinant proteins into culture was analyzed before and 24, 48, 72, 96 h and 6, 7, 10 and 12 days following irradiation. At the same time adherent cells were enumerated, evaluated for viability and proliferation. At 24, 48, 72 and 96 h postirradiation specific IL-6 and sIL-6R mRNA levels were analyzed.Irradiation of gene modified cells inhibited their proliferation in the dose dependant manner. Dose of 50 Gy sufficiently affected cell proliferation, however, for safety reasons we decided to use the dose of 100 Gy for vaccine preparation. Irradiation did not inhibit secretion of IL-6 and sIL-6R. In contrary, on a per cell basis it significantly increased their secretion which lasted 12 days postirradiation. Interestingly, we did not observe dose or time dependent differences in specific mRNA cellular levels suggesting that increased secretion of both proteins is regulated not on the transcriptional but rather on the posttranscriptional level. Taking all these facts into account we concluded that irradiation of tumor cells may provide an effective and safe approach for gene-modified vaccine preparation

47. Allogeneic bone marrow transplantation in children with acute lymphoblastic leukemia in first and second complete remission conditioned with fractionated total body irradiation and etoposide or cyclophosphamide

Patients and methodsFrom 1993 to 2001 thirty two children underwent bone marrow transplantation (BMT) for ALL (12 in I CR and 20 in II CR after early BM or BM/organ relapse). Except 2 syngeneic all other were HLA-identical siblings transplants. All patients (pts) were conditioned with FTBI 2×1,5 Gy for 4 days (total dose 12 Gy) with lung shielding (9 Gy) and CY 60 mg/kg i.v for 2 days (total dose 120 mg/kg) (n=1 in I CR and n = 11 in II CR) or VP 60 mg/kg i.v (n = 11 in I CR and n = 9 in II CR), Pts in I CR have been given 1,1–4,9×108 MNC/kg (med. 2,7×108/kg), while pts in II CR 1,9–4,0×108 MNC/kg (med. 2,7×108/kg). For GvHD prevention CsA 3 mg/kg/d i.v was administered alone in 22 pts (n = 9 in I CR and n = 13 in II CR) or in combination with “short” MTX +/− PRED in 8 pts (n = 3 in I CR and n = 5 in II CR). Two pts transplanted with syngeneic BM received no GvHD prevention. Regimen related toxicity (RRT) was graded according to the system developed by Bearman et al. (1988).ResultsOnly mild or moderate expression of RRT was observed (GI toxicity, I°− 80%, II° −4%; stomatitis I° −40%, II° −20%; hepatic toxicity I°− 28%; renal, bladder and cardiac toxicity I°− 4%) and no transplant related deaths occured (TRM=0%). Among 12 pts transplanted in I CR only one child relapsed 4 months from BMT, while remaining 11 pts are alive in CCR with a median follow-up of 33months (range 6 to 66 months) and 92% probability of 5-year EFS. Of 20 children transplanted in II CR 6 relapsed 1–14 months from BMT (median 6,5 months). Fourteen of them remain in CCR with a median follow-up 19,5 months (range 1 to 96 months) and 66% probability of 8-year EFS.Conclusions1.In children with ALL the FTBI-12 Gy-containing regimen is well tolerated without the life-threatening toxic complications.2.FTBI-12 Gy-containing regimen demonstrates very good antileukemic efficacy for HR-ALL in I CR, but only limited for ALL in II CA.3.3. In context of good tolerance of FTBI in a total dose of 12 Gy and its limited antileukemic efficacy in children with ALL in II CR the escalation of FTBI total dose from 12 Gy to 13,2 Gy appears to be justified in those children. Supported by grant KBN 4 PO5E 108 18

Age-Related Changes in the Natural Killer Cell Response to Seasonal Influenza Vaccination Are Not Influenced by a Synbiotic: a Randomised Controlled Trial

Natural killer (NK) cells are an important component of the immune response to influenza infection, but are subject to alteration during aging, which may play a role in impaired response to infection and vaccination in older people. Enhancement of NK cell activity could, therefore, present a means to improve the immune response to vaccination in older subjects, and pre- and probiotics offer an opportunity to modulate antiviral defenses via alteration of the gut microbiota. This study investigated the effect of a novel probiotic, Bifidobacterium longum bv. infantis CCUG 52486, combined with a prebiotic, gluco-oligosaccharide (B. longum + Gl-OS), on the NK cell response to seasonal influenza vaccination in young and older subjects in a double-blind, randomized controlled trial. There were significant effects of aging on NK cell phenotype, the most notable of which were an increase in CD56dim cells, mainly reflected in the CD16+ subset, a decrease in CD56bright cells, mainly reflected in the CD16− subset, and greater expression of the immunosenescence marker, CD57, on NK cell subsets. However, these changes only partially translated to differences in NK cell activity, observed as trends toward reduced NK cell activity in older subjects when analyzed on a per cell basis. Influenza vaccination increased the proportion of CD56bright cells and decreased the proportion of CD56dim cells, in young, but not older subjects. Although NK cell activity in response to vaccination was not significantly different between the young and older subjects, low post-vaccination NK cell activity was associated with poor seroconversion in only the older subjects. There was no influence of the synbiotic on NK cell phenotype or activity, either before or after influenza vaccination. In conclusion, aging is associated with marked alteration of the phenotype of the NK cell population and there was evidence of an impaired NK cell response to influenza vaccination in older subjects. The effects of aging on NK cell phenotype and activity could not be offset by B. longum + Gl-OS

Impact of ageing and a synbiotic on the immune response to seasonal influenza vaccination; a randomised controlled trial

Background & aims Ageing increases risk of respiratory infections and impairs the response to influenza vaccination. Pre- and pro-biotics offer an opportunity to modulate anti-viral defenses and the response to vaccination via alteration of the gut microbiota. This study investigated the effect of a novel probiotic, Bifidobacterium longum bv. infantis CCUG 52486, combined with a prebiotic, gluco-oligosaccharide, on the B and T cell response to seasonal influenza vaccination in young and older subjects . Methods In a double-blind, randomized controlled trial, 58 young (18–35 y) and 54 older (60–85 y) subjects were supplemented with the synbiotic for 8 weeks. At 4 weeks they were administered with a seasonal influenza vaccine. B and T cell phenotype and responsiveness to in vitro re-stimulation with the vaccine were assessed at baseline, 4, 6 and 8 weeks. Results B and T cell profiles differed markedly between young and older subjects. Vaccination increased numbers of memory, IgA+ memory, IgG+ memory and total IgG+ B cells in young subjects, but failed to do so in older subjects and did not significantly alter T cell subsets. Seroconversion to the H1N1 subunit in the older subjects was associated with higher post-vaccination numbers of plasma B cells, but seroconversion was less consistently associated with T cell phenotype. B and T cell subsets from both young and older subjects demonstrated a strong antigen-specific recall challenge, and although not influenced by age, responsiveness to the recall challenge was associated with seroconversion. In older subjects, CMV seropositivity was associated with a significantly lower recall response to the vaccine, but the synbiotic did not affect the responsiveness of B or T cells to re-stimulation with influenza vaccine. Conclusions Antigen-specific B and T cell activation following an in vitro recall challenge with the influenza vaccine was influenced by CMV seropositivity, but not by a synbiotic