Giovanni Sacchi

Acute Renal Failure from Inhalation of Mycotoxins

Mysterious deaths of archeologists after opening Egyptian tombs have been suspected to be secondary to inhalation of mycotoxin, however, the hypothesis has never been verified. Recently, we observed a case of acute renal failure (ARF) undeniably due to inhalation of ochratoxin of Aspergillus ochraceus. After spending 8 h in a granary which had been closed for several months, a farmer and his wife suffered temporary respiratory distress; 24 h later, the woman developed nonoliguric ARF and biopsy revealed tubulonecrosis which healed in 24 days. Toxic substances were not found, but a strain of A. ochraceus producing ochratoxin was isolated from the wheat.

The structure of superficial lymphatics in the human thigh: precollectors

Little is known about the morphology of precollectors, the lymphatic vessels connecting the absorbing and the collecting vessels, which are regarded as the initial drainage routes of lymph. The aim of this study was to describe the structural features of human precollectors.

Antiproliferative and Survival Properties of PMA in MCF-7 Breast Cancer Cell

Although PKCs are assumed to be the main targets of phorbol esters (PMA), additional PMA effectors, such as chimaerins (a family of RacGTPase activating proteins) and RasGRP (exchange factor for Ras/Rap1), can counteract or strengthen the PKC pathways. In this study, we evaluated the proliferative behavior of PMA-treated MCF-7 breast cancer cell and found that: PMA induced growth arrest and inhibited cell death; PMA activated ERKs, which, in turn, induced p21; and inhibitors of ERK (PD98059) and PKC (GF109203X) prevented p21 induction and abolished the PMA survival effect. We conclude that PMA inhibits MCF-7 cell growth and simultaneously stimulates cell survival; both responses are linked to ERK-dependent and p53-independent p21 induction.

Prevention of peritoneal sclerosis: a new proposal to substitute glucose with carnitine dialysis solution (biocompatibility testing in vitro and in rabbits).

Aim Commercial glucose peritoneal dialysis solutions expose the peritoneum to hyperosmolar glucose containing variable amounts of non-enzymic breakdown products of glucose. These solutions are toxic for the peritoneum. The aim of the present study is to compare in vitro and in vivo characteristics of a new dialysis solution containing carnitine, a naturally occurring compound, as substitute of glucose. Material and Methods We compared in vitro and in the rabbit a new peritoneal dialysis solution containing carnitine, with two standard bicarbonate glucose peritoneal dialysis solutions and a solution containing icodextrin. Results In vitro and in vivo the solution containing carnitine seems to be more biocompatible than standard glucose solutions and those containing icodextrin. Conclusions In our study the peritoneal dialysis solution containing carnitine seems to prevent the mesothelial changes observed with solutions containing glucose. Since carnitine has been extensively studied and seems to be well tolerated by hemodialysis patients, even at high doses for long periods, clinical trials in humans may be planned in the near future.

Simple peritoneal sclerosis and sclerosing peritonitis: related or distinct entities?

Aim The etiopathogenesis of sclerosing peritonitis is still debated, with some sustaining that it is a rare form of progression of simple peritoneal sclerosis and others that it is a primitive form. The aim of the present research was to clarify this question. Material and Methods 438 peritoneal biopsies from 253 patients were re-examined. 174 were obtained prior to peritoneal dialysis and 224 after various periods of dialysis. Forty biopsies were from peritoneal dialysis patients who developed sclerosing peritonitis. Peritoneal morphology was studied for signs of transition from simple sclerosis to sclerosing peritonitis. Results Evidence was found sustaining the hypothesis that simple sclerosis to sclerosing peritonitis patients have distinct pathologies. Conclusions The results confirm previous observations, excluding the existence of any type of relation between simple peritoneal sclerosis to sclerosing peritonitis.

Correction of erythrocyte shape abnormalities in familial hypercholesterolemia after LDL-apheresis: Does it influence cerebral hemodynamics?

SummaryIt is well known that red blood cells incubated in low-density lipoprotein (LDL)-rich medium show shape abnormalities that revert to normal after reincubation in normal plasma. Patients with homozygous familial hypercholesterolemia (HFH) have an increased percentage of abnormally-shaped erythrocytes (mostly stomatocytes, knisocytes, and crenated cells) compared to normocholesterolemic controls: 7.73±0.96 versus 3.52±0.52 (mean±SEM;P=0.001). To confirm the role of high LDL concentration in inducing red cell shape abnormalities we determined the percentage of abnormally shaped erythrocytes in seven HFH patients 1 day after the procedure of LDL-apheresis with a 40% cholesterol decrease. A reduction in kniscocytes, stomatocytes, and crenated cells was observed in the patients treated by LDL-apheresis (P<0.01).To investigate the possible benefit of a reduction in erythrocyte shape abnormality on cerebral hemodynamics, cerebral flow velocity, as evaluated by transcranial Doppler, was evaluated concommitanlly and found to be remarkably increased after apheresis (P<0.01). No significant change in hematocrit, plasma viscosity, blood viscosity, mean pressure, or cardiac output was detected, 1 day after apheresis. An inverse correlation was demonstrated (r=0.55;P=0.04) between changes in the percentage of knisocytes+stomatocytes+crenated cells and percent changes in middle cerebral artery peak systolic velocity. The correction of erythrocyte shape abnormalities after LDL-apheresis might be related to dramatic changes in plasma phospholipid concentration and proportion occurring after this procedure in HFH patients. The reduction of erythrocyte shape abnormalities could contribute, together with other hemorheological factors, to the improvement of cerebral hemodynamics after LDL-apheresis.

Culture of bovine thoracic duct endothelial cells

Dear Editor: In vitro studies have led to substantial advances in our understanding of the biology and physiologic role of blood vessel endothehum. The study of lymphatic endothelial cells (LEC) has been delayed due to the technical problems of isolation. LEC have been successfully isolated from bovine mesenteric lymphatic collectors (3-5,8), from canine thoracic duct (2), from a retroperitoneal recurrent lymphangioma (7), and more recently, from rat thoracic duct (1). LEC have also been isolated from human thoracic duct but these cells failed to grow in culture (2). The number of lymphatic endothelial cells that can be obtained from bovine mesenteric lymphatics and from the thoracic duct of dog and rat is, however, quite low due to the small size of these vessels, and insufficient for most biochemical studies. Here we report a reliable and easily reproducible technique for the isolation of a relatively large number of LEC from the main bovine lymphatic vessel, the thoracic duct. They may be utilized for biochemical investigations or other studies that require a high yield of early passage cells. Bovine thoracic aortas were obtained at the local slaughterhouse. The caudal extremity of the thoracic duct was identified in the periadventitial fat on the posterolateral side of the aortas caudal end, and injected with 0.1% Evans blue in saline (Fig. 1). As the thoracic duct, thus stained, was freed from most of the periadventitial fat, numerous small lymph nodes appeared. The lymph nodes never interrupted the course of the vessel, being connected with its lumen via short, thin collectors. The thoracic duct had no collateral branches; however, either of its extremities or the entire duct were sometimes duplicated. Both the extremities of the duct were cannulated with intravenous eannulae and the caudal extremity was dearly marked so that fluids could be infused in the normal direction of the lymph. The presence of valves prevented retrograde flow. The vessel was washed with phosphate buffered saline, filled with warm medium M199 (GIBCO, Life Technologies Ltd., Paisley, Scotland, UK) and incubated for 15 min at 37 ° C in a beaker containing warm saline. The vessel was then filled with 0.2% collagenase (Boehringer Mannheim, GmbH, W. Germany, 0.215 U/ mg) and incubated for 15 min at 37 ° C. After collecting the collagenase solution from the duct, the vessel was massaged to favor the detachment of LEC and washed with 30 ml of M199. All the effluents were spun for 10 min at 630 Xg. The cell pellet was resuspended in complete tissue culture medium (4): M199 containing 20% fetal bovine serum (GIBCO), 100 /.tg/ml endothelial ceil growth supplement (Sigma, St. Louis, MO), 100 /.tg/ml heparin (Sigma), 50 U/ml penicillin and 50 #g/ml streptomycin (GIBCO). The cells were seeded at a density of 1.6 × 104 cells/cm 2 into gelatin-coated plates or round glass cover slips, depending on intended use, and incubated in a humidified atmosphere with 5% CO2. The medium was changed 48 h after seeding and every other

State of the art on autologous mesothelial transplant in animals and humans.

Sixteen years ago rabbit and human mesothelial cells were successsfully cultured and autoimplanted. The aim of the study was merely to demostrate that mesothelial implant was possible and interesting not only in peritoneal dialysis, but also in the vaster field of medicine and surgery concerning all the mesothelial districts of the body. The aim of this paper is to recollect the steps which have led to autolougous mesothelial transplantation and verify if the tecnique has been validated and adopted by others. Review of the literature published in the last 15 years shows that intraperitoneal transplantation of mesothelial cells has been effective in reducing the formation of peritoneal adhesions, and in remodeling the area of mesothelial denudation. New studies on the mesothelial cell opened the way to costruction of transplantable tissue-engineered artificial peritoneum, to the utilization of mesothelial progenitor cells and to find simple metods to collect autologous mesothelial cells. Finally mesothelial trasnsplantation may represent a new neovascular therapy in the prevention and treatment of ischemic coronaric heart disease.

Experimental evaluation of transport force in the rabbit ureter

Background Cleaning the urinary tract by so-called “wash-out effect” and promoting high diuresis has long been advocated but has had very little scientific backing and few prospective studies in international journals. Aim To verify whether the physical laws describing the transport force of water in rivers and pipes are also valid for urinary outflow. Methods A laboratory model for measuring transport force, given liquid and solid capacity, was adapted to create an in vivo model based on the rabbit urinary tract. Results Fluid flow in the rabbit renal pelvis and ureters was found similar to flow in pipes, obeying the physical laws of water transport to some extent. When the quantity of liquid flowing in the urinary tract in unit time was doubled, the transport force increased by various orders of magnitude. When the liquid increased by a larger factor, the transport force became enormous. Conclusions The results confirm the utility of maintaining high diuresis in patients with renal calculus, but stress the utility of drinking 1–2 liters of hypotonic water in a short time to obtain an enormous increase in transport force which increases the probability of a cleansing effect.

A Novel Monoclonal Antibody Specific for Lymphatic Endothelium

The difficulty of identifying and differentiating lymphatic and blood microvessels in tissue sections can be overcome by a monoclonal antibody specific for lymphatic endothelium. Unfortunately, the only known antibody also reacts with the endothelium of some blood vessels. The technique of double immunization (passive, with an antiserum to blood endothelium, and active, with a suspension of lymphatic endothelial cells) was, therefore, used to increase the chances of recognizing specific lymphatic antigens by the mouse immune system. The monoclonal antibody obtained, LyMAb, a G1 immunoglobulin, reacted strongly with the endothelium of bovine thoracic duct, mesenteric collecting vessels and lymphatic vessels of gall-bladder and lymph nodes and moderately with those of the intestinal wall. Blood vessels (intercostal arteries, azygos vein and blood microvessels of all organs tested) were consistently negative. The antibody was species-specific and did not react with formalin-fixed, paraffin-embedded sections. Cross-reactivity was limited to some connective tissue fibres and scattered cells in the lymph node parenchyma, intestinal villi and hepatic lobules.