Immunohistochemistry detected and localized cannabinoid receptor type 2 in bovine fetal pancreas at late gestation

At present, data on the endocannabinoid system expression and distribution in the pancreatic gland appear scarce and controversial as descriptions are limited to humans and laboratory animals. Since the bovine pancreas is very similar to the human in endocrine portion development and control, studies on the fetal gland could prove to be very interesting, as an abnormal maternal condition during late pregnancy may be a predisposing trigger for adult metabolic disorders. The present investigation studied cannabinoid receptor type 2 presence and distribution in the bovine fetal pancreas towards the end of gestation. Histological analyses revealed numerous endocrinal cell clusters or islets which were distributed among exocrine adenomeri in connectival tissue. Immunohistochemistry showed that endocrine-islets contained some CB2-positive cells with a very peculiar localization that is a few primarily localized at the edges of islets and some of them also scattered in the center of the cluster. Characteristically, also the epithelium of the excretory ducts and the smooth muscle layers of the smaller arteries, in the interlobular glandular septa, tested positive for the CB2 endocannabinoid receptor. Conse quently, the endocannabinoid system, via the cannabinoid receptor type 2, was hypothesized to play a major role in controlling pancreas function from normal fetal development to correct metabolic functioning in adulthood

The presence and localization of apelin in the sheep abomasum: impact of diets characterized by different chemical composition

The apelinergic system is a complex system including the apelin peptide (AP), initially isolated in 1998 from bovine stomach homogenates and its receptor, named APJ receptor. The apelin peptide is extensively expressed in several human and laboratory animal organs (heart, lung, brain, mammary gland). Between the different roles hypothesize for apelin there are also the control of blood pressure and the stimulation of drinking behavior in rats. Recently its expression was also evidenced in the basal glandular portion of the stomach in laboratory animals, allowing to hypothesize its intervention in the control of acid secretion. No data concerning its presence and distribution in the abomasums of the sheep are present at the moment. So, we decided to test the presence and distribution of apelin in the abomasums of the sheep and the possible existence of a variability as a consequence of the different water content of the diet.The experiment was conducted using 50 sheep fed on Apennine semi-natural pasture for two experimental periods: in the first period the animals were fed for 45 days on the pasture at the maximum of its flowering; in the second one the animals were fed on the same pasture until its maximum dryness but the half of them daily received adiet integration of cereals. At the end of each experimental period, some animals were regularly slaughtered at the local abattoir, the abomasum specimens were immediately removed and some of them processed for routine tissue-embedding preparation while other for molecular biology. The immunohistochemical reaction was visualized on 5 µm serial sections, using a primary rabbit polyclonal antibody (anti-AP), the avidin-biotin-complex and the DAB as the chromogen. The immunohistochemical study showed a peculiar immunoreaction for AP in the abomasum of the animals examined. In particular, a immunopositive reaction for AP was evident in the cells of the basal third of the tubular glands and they were mainly of the closed type, with an oval or round shape and contained many perinuclear granules. The immunopositive reactions didn’t evidence any difference both in the localization and in the number of the positive cells, between the different groups. Immuno-positivity for AP was not observed in any other histological structure or in the sections utilized as negative controls.These results allow us to conclude that AP is present in the glandular tissue of the abomasums of the sheep, as observed in laboratory animals, with a peculiar cytoplasmatic localization and to hypothesize that AP is involved in the control of gastric secretion, probably via APJ receptor. The AP expression in the stomach doesn’t seem to be influenced by the water content of the diets with a positivity that appears to be localized in the same glandular portion between the different animal groups

ITALIAN CANCER FIGURES - REPORT 2015: The burden of rare cancers in Italy = I TUMORI IN ITALIA - RAPPORTO 2015: I tumori rari in Italia

OBJECTIVES: This collaborative study, based on data collected by the network of Italian Cancer Registries (AIRTUM), describes the burden of rare cancers in Italy. Estimated number of new rare cancer cases yearly diagnosed (incidence), proportion of patients alive after diagnosis (survival), and estimated number of people still alive after a new cancer diagnosis (prevalence) are provided for about 200 different cancer entities. MATERIALS AND METHODS: Data herein presented were provided by AIRTUM population- based cancer registries (CRs), covering nowadays 52% of the Italian population. This monograph uses the AIRTUM database (January 2015), which includes all malignant cancer cases diagnosed between 1976 and 2010. All cases are coded according to the International Classification of Diseases for Oncology (ICD-O-3). Data underwent standard quality checks (described in the AIRTUM data management protocol) and were checked against rare-cancer specific quality indicators proposed and published by RARECARE and HAEMACARE (www.rarecarenet.eu; www.haemacare.eu). The definition and list of rare cancers proposed by the RARECAREnet "Information Network on Rare Cancers" project were adopted: rare cancers are entities (defined as a combination of topographical and morphological codes of the ICD-O-3) having an incidence rate of less than 6 per 100,000 per year in the European population. This monograph presents 198 rare cancers grouped in 14 major groups. Crude incidence rates were estimated as the number of all new cancers occurring in 2000-2010 divided by the overall population at risk, for males and females (also for gender-specific tumours).The proportion of rare cancers out of the total cancers (rare and common) by site was also calculated. Incidence rates by sex and age are reported. The expected number of new cases in 2015 in Italy was estimated assuming the incidence in Italy to be the same as in the AIRTUM area. One- and 5-year relative survival estimates of cases aged 0-99 years diagnosed between 2000 and 2008 in the AIRTUM database, and followed up to 31 December 2009, were calculated using complete cohort survival analysis. To estimate the observed prevalence in Italy, incidence and follow-up data from 11 CRs for the period 1992-2006 were used, with a prevalence index date of 1 January 2007. Observed prevalence in the general population was disentangled by time prior to the reference date (≤2 years, 2-5 years, ≤15 years). To calculate the complete prevalence proportion at 1 January 2007 in Italy, the 15-year observed prevalence was corrected by the completeness index, in order to account for those cancer survivors diagnosed before the cancer registry activity started. The completeness index by cancer and age was obtained by means of statistical regression models, using incidence and survival data available in the European RARECAREnet data. RESULTS: In total, 339,403 tumours were included in the incidence analysis. The annual incidence rate (IR) of all 198 rare cancers in the period 2000-2010 was 147 per 100,000 per year, corresponding to about 89,000 new diagnoses in Italy each year, accounting for 25% of all cancer. Five cancers, rare at European level, were not rare in Italy because their IR was higher than 6 per 100,000; these tumours were: diffuse large B-cell lymphoma and squamous cell carcinoma of larynx (whose IRs in Italy were 7 per 100,000), multiple myeloma (IR: 8 per 100,000), hepatocellular carcinoma (IR: 9 per 100,000) and carcinoma of thyroid gland (IR: 14 per 100,000). Among the remaining 193 rare cancers, more than two thirds (No. 139) had an annual IR <0.5 per 100,000, accounting for about 7,100 new cancers cases; for 25 cancer types, the IR ranged between 0.5 and 1 per 100,000, accounting for about 10,000 new diagnoses; while for 29 cancer types the IR was between 1 and 6 per 100,000, accounting for about 41,000 new cancer cases. Among all rare cancers diagnosed in Italy, 7% were rare haematological diseases (IR: 41 per 100,000), 18% were solid rare cancers. Among the latter, the rare epithelial tumours of the digestive system were the most common (23%, IR: 26 per 100,000), followed by epithelial tumours of head and neck (17%, IR: 19) and rare cancers of the female genital system (17%, IR: 17), endocrine tumours (13% including thyroid carcinomas and less than 1% with an IR of 0.4 excluding thyroid carcinomas), sarcomas (8%, IR: 9 per 100,000), central nervous system tumours and rare epithelial tumours of the thoracic cavity (5%with an IR equal to 6 and 5 per 100,000, respectively). The remaining (rare male genital tumours, IR: 4 per 100,000; tumours of eye, IR: 0.7 per 100,000; neuroendocrine tumours, IR: 4 per 100,000; embryonal tumours, IR: 0.4 per 100,000; rare skin tumours and malignant melanoma of mucosae, IR: 0.8 per 100,000) each constituted <4% of all solid rare cancers. Patients with rare cancers were on average younger than those with common cancers. Essentially, all childhood cancers were rare, while after age 40 years, the common cancers (breast, prostate, colon, rectum, and lung) became increasingly more frequent. For 254,821 rare cancers diagnosed in 2000-2008, 5-year RS was on average 55%, lower than the corresponding figures for patients with common cancers (68%). RS was lower for rare cancers than for common cancers at 1 year and continued to diverge up to 3 years, while the gap remained constant from 3 to 5 years after diagnosis. For rare and common cancers, survival decreased with increasing age. Five-year RS was similar and high for both rare and common cancers up to 54 years; it decreased with age, especially after 54 years, with the elderly (75+ years) having a 37% and 20% lower survival than those aged 55-64 years for rare and common cancers, respectively. We estimated that about 900,000 people were alive in Italy with a previous diagnosis of a rare cancer in 2010 (prevalence). The highest prevalence was observed for rare haematological diseases (278 per 100,000) and rare tumours of the female genital system (265 per 100,000). Very low prevalence (<10 prt 100,000) was observed for rare epithelial skin cancers, for rare epithelial tumours of the digestive system and rare epithelial tumours of the thoracic cavity. COMMENTS: One in four cancers cases diagnosed in Italy is a rare cancer, in agreement with estimates of 24% calculated in Europe overall. In Italy, the group of all rare cancers combined, include 5 cancer types with an IR>6 per 100,000 in Italy, in particular thyroid cancer (IR: 14 per 100,000).The exclusion of thyroid carcinoma from rare cancers reduces the proportion of them in Italy in 2010 to 22%. Differences in incidence across population can be due to the different distribution of risk factors (whether environmental, lifestyle, occupational, or genetic), heterogeneous diagnostic intensity activity, as well as different diagnostic capacity; moreover heterogeneity in accuracy of registration may determine some minor differences in the account of rare cancers. Rare cancers had worse prognosis than common cancers at 1, 3, and 5 years from diagnosis. Differences between rare and common cancers were small 1 year after diagnosis, but survival for rare cancers declined more markedly thereafter, consistent with the idea that treatments for rare cancers are less effective than those for common cancers. However, differences in stage at diagnosis could not be excluded, as 1- and 3-year RS for rare cancers was lower than the corresponding figures for common cancers. Moreover, rare cancers include many cancer entities with a bad prognosis (5-year RS <50%): cancer of head and neck, oesophagus, small intestine, ovary, brain, biliary tract, liver, pleura, multiple myeloma, acute myeloid and lymphatic leukaemia; in contrast, most common cancer cases are breast, prostate, and colorectal cancers, which have a good prognosis. The high prevalence observed for rare haematological diseases and rare tumours of the female genital system is due to their high incidence (the majority of haematological diseases are rare and gynaecological cancers added up to fairly high incidence rates) and relatively good prognosis. The low prevalence of rare epithelial tumours of the digestive system was due to the low survival rates of the majority of tumours included in this group (oesophagus, stomach, small intestine, pancreas, and liver), regardless of the high incidence rate of rare epithelial cancers of these sites. This AIRTUM study confirms that rare cancers are a major public health problem in Italy and provides quantitative estimations, for the first time in Italy, to a problem long known to exist. This monograph provides detailed epidemiologic indicators for almost 200 rare cancers, the majority of which (72%) are very rare (IR<0.5 per 100,000). These data are of major interest for different stakeholders. Health care planners can find useful information herein to properly plan and think of how to reorganise health care services. Researchers now have numbers to design clinical trials considering alternative study designs and statistical approaches. Population-based cancer registries with good quality data are the best source of information to describe the rare cancer burden in a population

Lancet

BACKGROUND: In 2015, the second cycle of the CONCORD programme established global surveillance of cancer survival as a metric of the effectiveness of health systems and to inform global policy on cancer control. CONCORD-3 updates the worldwide surveillance of cancer survival to 2014. METHODS: CONCORD-3 includes individual records for 37.5 million patients diagnosed with cancer during the 15-year period 2000-14. Data were provided by 322 population-based cancer registries in 71 countries and territories, 47 of which provided data with 100% population coverage. The study includes 18 cancers or groups of cancers: oesophagus, stomach, colon, rectum, liver, pancreas, lung, breast (women), cervix, ovary, prostate, and melanoma of the skin in adults, and brain tumours, leukaemias, and lymphomas in both adults and children. Standardised quality control procedures were applied; errors were rectified by the registry concerned. We estimated 5-year net survival. Estimates were age-standardised with the International Cancer Survival Standard weights. FINDINGS: For most cancers, 5-year net survival remains among the highest in the world in the USA and Canada, in Australia and New Zealand, and in Finland, Iceland, Norway, and Sweden. For many cancers, Denmark is closing the survival gap with the other Nordic countries. Survival trends are generally increasing, even for some of the more lethal cancers: in some countries, survival has increased by up to 5% for cancers of the liver, pancreas, and lung. For women diagnosed during 2010-14, 5-year survival for breast cancer is now 89.5% in Australia and 90.2% in the USA, but international differences remain very wide, with levels as low as 66.1% in India. For gastrointestinal cancers, the highest levels of 5-year survival are seen in southeast Asia: in South Korea for cancers of the stomach (68.9%), colon (71.8%), and rectum (71.1%); in Japan for oesophageal cancer (36.0%); and in Taiwan for liver cancer (27.9%). By contrast, in the same world region, survival is generally lower than elsewhere for melanoma of the skin (59.9% in South Korea, 52.1% in Taiwan, and 49.6% in China), and for both lymphoid malignancies (52.5%, 50.5%, and 38.3%) and myeloid malignancies (45.9%, 33.4%, and 24.8%). For children diagnosed during 2010-14, 5-year survival for acute lymphoblastic leukaemia ranged from 49.8% in Ecuador to 95.2% in Finland. 5-year survival from brain tumours in children is higher than for adults but the global range is very wide (from 28.9% in Brazil to nearly 80% in Sweden and Denmark). INTERPRETATION: The CONCORD programme enables timely comparisons of the overall effectiveness of health systems in providing care for 18 cancers that collectively represent 75% of all cancers diagnosed worldwide every year. It contributes to the evidence base for global policy on cancer control. Since 2017, the Organisation for Economic Co-operation and Development has used findings from the CONCORD programme as the official benchmark of cancer survival, among their indicators of the quality of health care in 48 countries worldwide. Governments must recognise population-based cancer registries as key policy tools that can be used to evaluate both the impact of cancer prevention strategies and the effectiveness of health systems for all patients diagnosed with cancer. FUNDING: American Cancer Society; Centers for Disease Control and Prevention; Swiss Re; Swiss Cancer Research foundation; Swiss Cancer League; Institut National du Cancer; La Ligue Contre le Cancer; Rossy Family Foundation; US National Cancer Institute; and the Susan G Komen Foundation

AN IMMUNOHISTOCHEMICAL STUDY EVIDENCES GHRELIN AND ITS RECEPTOR IN THE HAIR FOLLICLE OF THE SHEEP

Ghrelin is a peptide hormone discovered in 1999. It is mainly secreted by the endocrine cells of the gastrointestinal tract and acts by binding to a specific receptor. Ghrelin presents a wide tissue distribution and is involved in numerous central and peripheral actions including hormonal, orexigenic, neurological, cardiovascular, and immunological activities (1).In this work, the expression and localization of ghrelin and its receptor was investigated in the skin of sheep by means of immunohistochemistry. The immunohistochemical reaction was performed on ovine skin samples collected from the neck ventral region of healthy animals regularly slaughtered at the abattoir. Samples were fixed in formalin and embedded in paraffin. Dewaxed sections were incubated with 3% peroxidase-blocking solution and with normal goat serum to block the endogenous peroxidase activity and non-specific binding respectively. Successively, serial sections were incubated overnight with polyclonal anti-Ghrelin and anti-Ghrelin receptor antibodies (Abcam Cambridge UK). The reaction was visualized using Vectastain ABC kit and DAB (Vector Laboratories, Burlingame, CA, USA). The analyses performed evidenced a clear and intense immunostaining for both ghrelin and its receptor in the ovine skin. Staining was mainly localized in the hair follicles (HF). Positivity to ghrelin was observed in a short area of the HF at the level of the soprabulbar region; it involved the inner cells of the outer root sheath, including the companion layer, and the cells of the inner root sheath. The receptor, instead, was observed in all cell layers of the outer root sheath and extended more than ghrelin, from the soprabulbar region to the isthmus. Other than HF, ghrelin showed a weak positivity in the soprabasal layers of the epidermis which nevertheless were negative to the receptor. Finally, the receptor was clearly expressed by the cells of the sweat glands. At present, the skin is described as an endocrine organ since it is the target of several endocrine signals and, at the same time, it is itself capable of producing substances with hormone-like activity (2). The study of ghrelin and, more in general, of adipokines at the skin level represent an interesting and current topic for domestic animals including sheep. It was shown that some adipokines, such as leptin and adiponectin, are widely involved in the metabolism of the skin, and, more specifically, of the HF in both physiological and pathological conditions (3, 4). Ghrelin is a recently discovered molecule and there are until now a few surveys on ghrelin at the skin level. However the strong immunohistochemical expression of ghrelin and its receptor evidenced in the HF of the sheep let us to suppose that ghrelin may have a role in the HF activity probably acting through a paracrine or autocrine mechanism

EPIDERMIS AND HAIR FOLLICLE IN BOVINE SKIN EXPRESS THE LEPTIN HORMONE AND ITS RECEPTOR

Leptin (Ob) is a hormone that acts through the activation of the specific receptor Ob-R. It is mainly secreted by visceral and subcutaneous adipose tissue and represents the first known adipokine [1]. Ob is a pleiotropic molecule and plays an important role in the skin, where it stimulates keratinocytes to proliferate and intervenes in the regulation of wound healing processes. It also participates in the control of hair follicle morphogenesis and its cycles of growth, regression, and rest [2]. Ob may act through endocrine, paracrine and autocrine mechanisms. While it is secreted by skin structures including epidermis and hair follicles, intradermal adipose tissue also seems to have a role in Ob secretion and accordingly in the control of the hair follicle cycle in mice and humans [3]. In this work, the morphological characteristics of the skin in bovine species were evaluated by Hematoxylin-Eosin and Oil Red O stainings to investigate the presence and extension of intradermal adipose tissue that may be invoved in Ob secretion. Moreover, Ob and Ob-R expression were analized by means of RT-PCR and immunohistochemistry. Through the morphological analysis, a high and thick dermis without adipocytes was observed. Hair follicles and glands were located in the prossimal part of the skin, beneath the epidermis, while a thick layer of connective tissue, lacking adipose cells, separated these structures by subcutis. RT-PCR gave a positive outcome, evidencing the transcripts for both molecules in the bovine skin samples examined By immunohistochemistry, Ob and its receptor were observed in the epidermis and in the outer root sheath of hair follicles during the follicular cycle. The epidermis abundantly expressed Ob; while all layers of cells were involved, the suprabasal layers expressed a stronger signal. Ob-R was observed in the cells of the basal layer. As regards hair follicles, both Ob and Ob-R were expressed by the outer root sheath of hair follicles. Staining mainly extended into the regions of the infundibulum and isthmus while the bulb was negative. Immunostaining persisted in all stages of hair follicles. The expression of Ob-R in the bovine skin proves that Ob acts on this peripheral organ. The identification of Ob in the epidermis and hair follicle epithelium attests that Ob may act through a paracrine and autocrine mechanism on these structures even if an endocrine mechanism cannot be excluded. The absence of adipocytes around hair follicles and, broadly, in all the dermis indicates that the intradermal adipose tissue does not exist in bovine and accordingly cannot exert paracrine control on the hair follicle. The identification of the Ob system in bovine skin provides important information for properly understanding the biological mechanisms that regulate skin structures, and well as for comparing animal species and highlighting their differences