MATER protein as substrate of PKCepsilon in human cumulus cells.

High activity of the phosphoinositide 3-kinase/Akt pathway in cumulus cells plays an important role in FSH regulation of cell function and Protein Kinase C epsilon (PKCepsilon) collaborates with these signalling pathways to regulate cell proliferation. Relevant roles in follicular development are played by Maternal Antigen That Embryos Require (MATER) that is a cumulus cell- and oocyte-specific protein dependent on the maternal genome. We recently demonstrated that human MATER localizes at specific domains of oocytes and, for the first time, also in cumulus cells. MATER contains a carboxy-terminal leucine-rich repeat domain involved in protein-protein interactions regulating different cellular functions. Here we investigated the functional role of MATER. Thus, we performed coimmunoprecipitation experiments using HEK293T cells expressing human MATER; a similar approach was then followed in human cumulus/follicular cells. In MATER(+)HEK293T cells, we observed that this protein acts as a phosphorylation substrate of PKCepsilon. Western blot experiments indicate that, unlike oocytes, human cumulus cells express PKCepsilon. Immunoprecipitation and confocal analysis suggest for the first time that MATER protein interacts with this protein kinase in cumulus cells under physiological conditions. Since PKCepsilon is known to collaborate with antiapoptotic signalling pathways, this suggests a novel mechanism for the function of MATER in follicular maturation.

Aberrant crypt foci in colorectal carcinogenesis. Cell and crypt dynamics.

Aberrant crypt foci (ACF) have been identified on the colonic mucosal surface of rodents treated with colon carcinogens and of humans after methylene-blue staining and observation under a light microscope. Several lines of evidence strongly suggest that ACF with certain morphological, histological, cell kinetics, and genetic features are precursor lesions of colon cancer both in rodents and in humans. Thus, ACF represent the earliest step in colorectal carcinogenesis. This paper has the main purpose of reviewing the evidence supporting this view, with particular emphasis on cell and crypt dynamics in ACF. ACF have been used as intermediate biomarkers of cancer development in animal studies aimed at the identification of colon carcinogens and chemopreventive agents. Recently, evidence has also shown that ACF can be effectively employed in chemopreventive studies also in humans.

Complications and long-term outcome of 80 oncology patients undergoing needle catheter jejunostomy placement for early postoperative enteral feeding.

Some findings suggest that needle catheter jejunostomy (NCJ) is associated with a significant rate of potentially dangerous complications. The purpose of this study was to prospectively evaluate the rate and type of early and late complications associated with NCJ in patients with surgical treatment of upper gastrointestinal malignancy. Eighty patients underwent NCJ implant at the end of their scheduled surgical procedure. Enteral nutrition programme was started on postoperative day 1 in the surgical ICU. NCJ was always removed in the outpatient clinic after hospital discharge. One case of tube blockage has been observed as single short-term complication in this series. No long-term complications have been detected after a mean follow-up of 12 months. Routine use of NCJ in malnourished patients undergoing major surgical procedures on upper gastrointestinal tract is safe and effective.

Apoptosis in different stages of human oogenesis.

Apoptosis is an active form of cell death characterized by a series of morphological changes that become particularly evident at the ultrastructural level. The majority of ovarian germ cells undergo degeneration during prenatal and reproductive life and only in recent studies has it been demonstred that this drop is due to an apoptotic process. We evaluated this process during human oogenesis in prenatal life and we studied the ultrastructural changes that occur in apoptosis in various phases of the meiotic process. From our observations it is clear that apoptosis involves two main phases of the meiotic process: an earlier one concerning the oogonia and oocytes in the preleptotene stage, and a later one that mainly concerns the oocytes in the pachytene stage.

Scanning electron microscopy of aberrant crypt foci in human colorectal mucosa.

BACKGROUND: Aberrant crypt foci (ACF) are clusters of morphologically altered crypts which can be observed by light or stereomicroscopy on the mucosal surface of the colon after staining with methylene-blue. They probably represent one of the earliest events in human colorectal carcinogenesis. The main purpose of the present study was to observe the surface features of aberrant and normal colonic crypts in humans using scanning electron microscopy (SEM) in order to find and measure differences between aberrant and normal. MATERIALS AND METHODS: Fifteen mucosal specimens containing ACF and 8 with normal mucosa taken from patients operated on for colon cancer were observed under a scanning electron microscope. RESULTS: By SEM ACF were easily observed on the mucosal surface, because they showed a well defined border and were elevated on the mucosal surface. Under higher magnification luminal openings of aberrant crypts had a larger overall average diameter than normal (37.6 microns +/- 13.5, mean +/- SD, vs 15.9 microns +/- 4.9, P = 0.001), though when crypt multiplicity of ACF (number of crypts per ACF) was higher, the diameter of luminal openings tended to be smaller and similar to those of normal crypts, with weak negative correlation between crypt multiplicity of ACF and mean diameter of aberrant luminal openings (r = 0.27). Finally, the mucosal surface among aberrant crypts was flattened because of a loss of microvilli. in conclusion, scanning electron microscopy allows a better definition of the topological features of aberrant crypt foci than light or stereomicroscopy.

Rare presentation of small bowel leiomyosarcoma with liver metastases.

Intraabdominal sarcomas are rare tumours usually diagnosed at an advanced stage. These lesions at presentation are bulky and symptoms are often related to pressure effects on adjacent organs. This case report describes a rare presentation of a small bowel leiomyosarcoma whose initial presentation was free haemorrhage into the abdominal cavity and concomitant liver metastases. This case report also demonstrates that, even with such a rare presentation, an aggressive surgical approach is indicated in this type of tumour and helps a patient with advanced disease to live a few disease-free months with a good quality of life.

Effects of estrogens and oxytocin on the development of neonatal mammalian ovary.

The preservation and death of germ cells in the neonatal mammalian ovary are linked with the presence of hormones. Estrogens and oxytocin are present at birth in all mammalian vertebrates. The aim of this study was to examine their role in the development of the neonatal ovary and also in the preservation and death of germ cells in the neonatal period: apoptotic phenomena play a fundamental role in the control of their number. Female neonatal mice were treated at birth with estradiol monobenzoate or oxytocin and sacrificed after 5 days. The ovaries were sectioned in toto into semi-thin sections, in order to calculate their volume. Thin sections were also carried out to verify, under the transmission electron microscope (T.E.M.), the cells in apoptosis. The ovaries treated with the greater concentration of estradiol monobenzoate showed a volume that was significantly greater than that of the controls and a reduction of germ cells in apoptosis. The ovaries treated with oxytocin at all degrees of concentration had a volume significantly less than the controls and they also had a higher number of germ cells in apoptosis.

Human neonatal ovary: proposal of a three-dimensional model.

A model of human neonatal ovary is presented, derived from morphometric, evaluations carried out on left ovaries removed from five full-term neonates with a 46, XX karyotype, free from malformations of the genital apparatus. According to this model, the gonad can be represented by a triaxial ellipsoid with a central medullary core surrounded by a cortical stratum of constant thickness. The germinal population, consisting of follicles and primitive cortical tissue, occupies the cortex, intermingled with the interstitium or stroma. In the cortex it is then possible to describe an outer layer formed by primitive cortical tissue, and an inner portion occupied by follicles. The primary and secondary follicles fill the portion near the medulla and the primordial ones are contained in the middle and outer zones. Since the variability observed among ovaries is slight, we can propose a mean model of neonatal ovary in which the spatial relationships among the different components, the total number of follicles and their position in the cortex can be calculated.

[An experimental study of the use of synthetic meshes in large abdominal eventrations]. / Studio sperimentale sull'applicazione di reti sintetiche in corso di grandi sventramenti addominali.

BACKGROUND: The aim of this study is to evaluate the biocompatibility of Angimesh mesh (polypropilene) in rats, usually used to replace or to strengthen the abdominal fasciae. METHODS: We made an abdominal longitudinal incision of skin and muscles in 20 rats was carried out and two muscular semilunae removed. The incision was occluded with Angimesh mesh fixed by ethilcyanoacrilate glue (first group of rats) or with suture stitches (second group of rats). The animals were sacrificed after 80 days and the stability of the mesh and the presence of endoperitoneal adhesion were evaluated macroscopically, and microscopically the fibrogenesis and inflammation answer. RESULTS: Two rats of the first group and seven of the second group died before 30 days. The mesh was more steady when fixed by suture. Adhesion in the contact sites between the mesh and the peritoneum was found. Histology did not show any inflammation reaction but showed a fibrogenic answer. CONCLUSIONS: The cyanoacrilate non-toxicity has been confirmed also in this application. Angimesh mesh showed good biocompatibility and resulted ideal for the synthesis of large abdominal resections.

[Artificial nutrition in the management of lesions caused by caustic ingestion]. / La nutrizione artificiale nelle lesioni da ingestione di caustici.

Ingestion of a large amount of corrosive agent results in a life-threatening condition which requires a much more aggressive diagnostic and therapeutic approach than was formerly recommended. Based on experience with a wide spectrum of upper g-i injuries in 56 patients treated through a 5-year-period, the Authors suggest an early staging of the lesions by immediate endoscopy, followed by adequate resective surgery for high-degree esophago-gastric lesions. Indications, techniques of artificial nutrition and the nutritional requirements of this condition are presented and fully discussed. The prompt institution of total parenteral nutrition is an essential part of the emergency management of these lesions. Fourteen patients underwent surgical treatment for the lesions; in all cases a Witzel's feeding jejunostomy was performed and postoperative enteral nutrition was started. Eight patients underwent a reconstructive operation of colon interposition and received home-based total enteral nutrition for a period of 8-25 weeks. No major complications relate to nutritional support occurred and nutritional indexes were normal at the end of the treatment.

Flexible sigmoidoscopy as a colorectal cancer screening test in the general population: recruitment phase results of a randomized controlled trial in Lombardia, Italy.

UNLABELLED: A two arm multicentre randomized controlled trial is in progress to evaluate the efficacy of flexible sigmoidoscopy (FS) as a screening test for colorectal cancer in the general population. AIMS: To determine the acceptance rate and feasibility of FS as a colorectal cancer screening test in average-risk asymptomatic volunteers. Average-risk, asymptomatic subjects, aged 55-64 years and assisted by 244 general practitioners (GPs) in Lombardy, Italy, were invited by postal questionnaire (PQ) to enter a study for the prevention of colorectal cancer and asked to indicate their interest in, and willingness to undergo, screening: those responding positively were randomized to the intervention or control arms. GPs were trained in colorectal cancer screening and proposed free FS to their patients randomized to the intervention arm. All sigmoidoscopies were performed by experienced endoscopists. Small polyps were removed at FS. Colonoscopy was indicated for high risk polyps (size more than 5 mm, more than two adenomas, villous histology, severe dysplasia or malignancy). 40,945 subjects were invited. 667 PQs were returned undelivered due to postal failure. 7,892 (19.59%) subjects responded, 2,116 of whom (26.81%) were not included, presenting 1 or more exclusion criteria. We randomized 5,778 volunteers and performed 1,582 sigmoidoscopies out of 2,885 subjects in the intervention arm (54.84% acceptance rate). Although the screening procedure had a good attendance rate in the intervention group, involvement of the people invited was lower than expected. Future FS screening programmes will require a keener focus on recruitment strategies, mainly with participation of GPs.

[Feasibility of exeresis. Controversial aspects in the surgery of carcinoma of the pancreas]. / Fattibilità della exeresi. Aspetti controversi nella chirurgia del carcinoma del pancreas.

Pancreatic cancer has a dismal prognosis also after resection with a 5 years' survival of about 5% in operated patients. The main clinical issue in patients with a malignant tumour is to identify the ones that would benefit from a surgical treatment. Resectability of pancreatic cancer has not an absolute value and the possible advantages in terms of prognosis and quality of life should be balanced with surgical mortality and morbidity. For this reason the management of this disease involves a multidisciplinary approach and the surgeon should join with the other specialists in experienced oncology centers. En exhaustive evaluation of the following prognostic factors should be made pre and intra-operatively to better define life expectancy with or without resection: Histotype: endocrine tumours and cystadenocarcinoma have, in general, a better prognosis, Staging: JPS classification has a better prognostic value if compared to the UICC. Completeness of the resection. Biological characteristics of the tumour. The main variables to be considered for the exeresis are: Size and local growth of the tumour (also considering the involvement of vessels, retroperitoneum and pancreatic capsule). Liver or peritoneal metastases: for this laparoscopy has a key role for staging. Histologic confirmation: differential diagnosis with chronic pancreatitis is sometimes difficult and every attempt should be made to have a pre-operative histology. Vascular invasion is one of the main contraindications to surgery and an exhaustive evaluation of vascular involvement should be considered mandatory. Lymph nodal involvement, in general, represents a negative prognostic factor even if Japanese authors claim that a radical resection can be performed in case of positive nodes in the peripancreatic area, if a complete lymphadenectomy is carried out.

Application of poly-L-lactide screws in flat foot surgery: histological and radiological aspects of bio-absorption of degradable devices.

The flat foot in childhood is a condition frequently observed in orthopedic practice but it is still debated when and in which patients surgical corrective treatment is appropriate; recently, the application of poly-L-lactic-acid (PLLA) screws was proposed. The present study investigates a group of 33 patients treated with PLLA expansion endorthesis in order to evaluate the deformity correction. Clinical and radiological outcomes in patients were correlated with: a) morphological characterization of screws both before and after being removed from patients, when necessary; b) histological and bio-molecular evaluation of degradation processes of the implants, focusing attention on the correlation between the cellular cohort involved in inflammatory reaction and the bio-absorption degree of PLLA screws. Deformity correction was mostly achieved, with minimal need of screw removal; the results obtained clearly show the occurrence of chronic rather than acute inflammation in removed screw specimens. At the histological level, after biomaterial implantation, the sequence of events occurring in the surrounding tissues ultimately ends in the formation of foreign body giant cells (FBGCs) at the tissue/material interface; but the mechanisms which influence the fate of screw implants, i.e. the resolution of acute inflammation rather than the progression towards chronic inflammation, are of crucial importance for biodegradable materials like "polylactic acid". In fact, the FBGC response ensures a long-term mechanism which eliminates the foreign material from the body, but at the same time the implications of prolonged FBGC responses, which generate negative side effects, could significantly impede the healing progress.

Effect of ethanol on respiratory activity of embryonal tissues.

By estimating the respiratory activity of various chicken embryonal organs we have noted that it was the highest for the kidney tissues and lower and lower for liver, brain and heart tissues. After the addition of ethanol to the culture medium we may conclude that ethanol 2,5% does not substantially alter the cellular respiratory activity, whereas ethanol 5% remarkably appears to inhibit it. In fact for this last condition the heart and kidney fragments, when are transplanted after 48 hours in "pendent drop", result rarely surviving.

Effect of organ phospholipids on the growth of embryonal tissues cultured in vitro.

With regard to a set of studies about cellular nutrition, the effect of different amounts of phospholipids, extracted from various organs of calf (diencephalon, retina, cerebral cortex and heart), on normal embryonal chicken myocardium cultures has been tested. From the numerous observations we have made it is resulted that phospholipids, particularly at the highest levels, constantly decrease the culture migration velocity and this reduction is remarkably more accentuated in presence of 1.25%(0) heart phospholipids. Moreover in this condition the cells show a change in their morphology and appear all full of big lipid drops, perhaps in consequence of an alteration of the chemical-physical plasmalemma characteristics and a change in the membrane enzyme activities involving lipid metabolism too.