Massive infection of Cystidicoloides ephemeridarum in brown trout Salmo trutta with skeletal deformities.

We investigated the cause of skeletal deformities found in brown trout from the Aspromonte mountain area in Reggio Calabria, Italy. Toxicological, histopathological and parasitological analyses were carried out on 14 fish with evident macro-morphological alterations from 2 different locations in the same river, and 4 control fish without morphological alterations from a different river (far from the first river but still within the area under study). Histopathological and radiological observations confirmed severe skeletal deformities in the specimens investigated. Parasitological examinations highlighted the presence of the nematode Cystidicoloides ephemeridarum, found only within the gastrointestinal tract of specimens showing deformities. Moreover, a direct correlation between parasite number and fish size was found. Given the low heavy metal levels and the presence of a massive parasitosis in teleosts showing deformities, we postulate a correlation between skeletal deformities and nematode infestation: the parasites caused a serious vitamin and mineral deficiency in the fish, which led to a dysplastic vertebral column. The low calcium levels found in malformed specimens compared with negative controls effectively confirm this hypothesis.

Gastric leiomyoma in a sea bass Dicentrarchus labrax broodfish.

This report describes a spontaneously arising non-infiltrative neoplasm of gastric tunica muscularis in a broodstock sea bass Dicentrarchus labrax from an Italian aquaculture farm. Microscopically, the mass was circumscribed and non-encapsulated and was composed of spindle cells arranged in parallel interlacing bundles or, occasionally, a whirling pattern. Cells had a small quantity of eosinophilic cytoplasm with distinct cell borders. Neoplastic cells were immuno-reactive with smooth muscle actin, vimentin and desmin; S100 was negative. The mucosal epithelium was intact with no neoplastic involvement. A gastric leiomyoma was diagnosed based on the findings. More efforts should be made to study the possible etiology of leiomyoma affecting fish from aquaculture.

Immunohistochemical Study of Four Fish Tumors.

The present study supports the usefulness of ancillary techniques, such as immunohistochemistry, as a valid diagnostic tool in the field of fish oncology. The immunohistochemical patterns observed in four neoplasms on four individual teleosts belonging to different species are described. Cytokeratin, vimentin, actin, S100, calretinin, and Melan-A antibodies were used. Diagnoses of papilloma in a Bream Abramis brama, fibroma in a Sand Steenbras Lithognathus mormyrus, schwannoma in a Crucian Carp Carassius carassius, and melanoma in a spontaneously inbred Xiphophorus hybrid were made. Diagnosis of tumors in fish is not always easy to carry out, and the tool provided by antibodies used on mammalian tissue is essential for obtaining definitive, unambiguous, and inexpensive identification.

Electrophysiological dysfunction induced by anti-ribosomal P protein antibodies injection into the lateral ventricle of the rat brain.

Objective Anti-ribosomal P antibodies (anti-P) are strongly associated with neuropsychiatric lupus. This study was designed to determine whether these antibodies are capable of causing electro-oscillogram (EOSG) and behavior alterations in rats. Methods IgG fraction anti-P positive and affinity-purified anti-P antibodies were injected intraventricularly in rats. Sequential cortical and subcortical EOSGs were analyzed during 30 days. IgG anti-Ro/SS-A and normal IgG were used as controls. Results All 13 animals injected with IgG anti-P demonstrated a high prevalence of polyspikes, diffusely distributed in hippocampal fields and cerebral cortex. These abnormalities persisted approximately a month. Remarkably, an identical electrical disturbance was observed with the inoculation of affinity-purified anti-P antibodies. The EOSG alterations were associated with behavioral disorders with varying degrees of severity in every animal injected with anti-P. In contrast, no changes in EOSG or behavioral disturbances were observed in the control group. Conclusion Our study indicates that anti-P antibodies can directly induce electrophysiological dysfunction in central nervous system particularly in hippocampus and cortex associated with behavior disturbances.

The role of endosomal toll-like receptors in bacterial recognition.

BACKGROUND: Infections caused by extracellular Gram positive bacteria are still a major health problems. Better understanding of the mechanisms underlying immune responses to these organisms is key to develop pharmacological agents, including vaccines, to control these infections. OBJECTIVE AND PERSPECTIVES: The objective of this review is to highlight the importance of nucleic acid-sensing, intracellular Toll-like receptors in innate immune recognition and in host defenses against extracellular bacteria. CONCLUSIONS: Toll-like receptors 7 and 9 have a major role in inducing host-protective type I interferon responses in conventional dendritic cells in response to streptococci and other extracellular gram positive bacteria. Moreover an as yet unidentified MyD88-dependent receptor is likely responsible for proinflammatory cytokine induction in response to these pathogens.

Haemophagocytic syndrome in rheumatic patients. A systematic review.

BACKGROUND: Hemophagocytic lymphohistiocytosis (HLH), is a potentially fatal hyperinflammatory syndrome characterized fever, hepatosplenomegaly, and cytopenias. HLH can be either primary, with a genetic aetiology, or secondary, associated with malignancies, autoimmune diseases, or infections. Among rheumatic disorders, HLH occurs most frequently in systemic juvenile idiopathic arthritis. AIM: To draw attention on this severe syndrome that may often go undiagnosed in patient with rheumatic diseases. MATERIALS AND METHODS: PubMed search was performed by combining the terms (haemophagocytic, haemophagocytosis, hemophagocytosis, hemophagocytic, erythrophagocytosis, macrophage activation syndrome) and (rheumatic, rheumatologic, arthritis, lupus, Sjögren's syndrome, scleroderma, polymyositis, dermatomyositis, polymyalgia rheumatic, mixed connective tissue disease, polychondritis, sarcoidosis, polyarteritis nodosa, Henoch-Schönlein, serum sickness, wegener's granulomatosis, giant cell arteritis, temporal arteritis, Takayasu's arteritis, Behçet's syndrome, Kawasaki, Buerger's). RESULTS: 117 papers describing 421 patients were considered. HLH was described in systemic lupus erythematosus in 94 patients, in Still's disease in 37 patients, in rheumatoid arthritis in 13 patients, in systemic juvenile arthritis in 219 patients, in dermatomyositis in 7 patients, in Kawasaki disease in 25 patients, in systemic sclerosis in 5 patients, in Behcet disease in one patient, in polyarteritis nodosa in 6 patients, in ankylosing spondylitis in 2 patients, in mixed connective tissue disease in one patient, in sarcoidosis in 5 patients, in Sjögren's syndrome in 3 patients, in Wegener's granulomatosis in one patient, and in unclassifiable disorders in two patients. CONCLUSIONS: HLH occurring in the course of rheumatic diseases is an important and often underdiagnosed clinical entity, which can affect prognosis.

Secondary hemophagocytic lymphohistiocytosis in zoonoses. A systematic review.

BACKGROUND: Hemophagocytic lymphohistiocytosis (HLH) is a rare syndrome that is often fatal despite treatment. It is caused by a dysregulation in natural killer T-cell function, resulting in activation and proliferation of histiocytes with uncontrolled hemophagocytosis and cytokines overproduction. The syndrome is characterized by fever, hepatosplenomegaly, cytopenias, liver dysfunction, and hyperferritinemia. HLH can be either primary, with a genetic aetiology, or secondary, associated with malignancies, autoimmune diseases, or infections. AIM: To focus on secondary HLH complicating zoonotic diseases. MATERIALS AND METHODS: PubMed search of human cases of HLH occurring during zoonotic diseases was performed combining the terms (haemophagocytic or haemophagocytosis or hemophagocytosis or hemophagocytic or erythrophagocytosis or macrophage activation syndrome) with each one of the etiological agents of zoonoses. RESULTS: Among bacterial diseases, most papers reported cases occurring during brucellosis, rickettsial diseases and Q fever. Regarding viral diseases, most of the cases were reported in patients with avian influenza A subtype H5N1. Among the protozoan zoonoses, most of the cases were reported in patients with visceral leishmaniasis. Regarding zoonotic fungi, most of the cases were reported in AIDS patient with histoplasmosis. No cases of secondary HLH were reported in patient with zoonotic helminthes. CONCLUSIONS: Zoonotic diseases are an important cause of HLH. Secondary HLH can delay the correct diagnosis of the zoonotic disease, and can contribute to an adverse outcome.

A mycobacterial disease is associated with the silent mass mortality of the pen shell Pinna nobilis along the Tyrrhenian coastline of Italy.

Disease is an increasing threat for marine bivalves worldwide. Recently, a mass mortality event (MME) impacting the bivalve Pinna nobilis was detected across a wide geographical area of the Spanish Mediterranean Sea and linked to a haplosporidian parasite. In 2017-2018, mass mortality events affecting the pen shell Pinna nobilis were recorded in two different regions of Italy, Campania and Sicily, in the Tyrrhenian Sea (Mediterranean Sea). Histopathological and molecular examinations of specimens showed the presence of Haplosporidium sp. in only one specimen in one area. Conversely, in all of the surveyed moribund animals, strong inflammatory lesions at the level of connective tissue surrounding the digestive system and gonads and linked to the presence of intracellular Zhiel-Neelsen-positive bacteria were observed. Molecular analysis of all of the diseased specimens (13) confirmed the presence of a Mycobacterium. Blast analysis of the sequences from all of the areas revealed that they were grouped together with the human mycobacterium M. sherrisii close to the group including M. shigaense, M. lentiflavum and M. simiae. Based on pathological and molecular findings, it is proposed that a mycobacterial disease is associated with the mortality episodes of Pinna nobilis, indicating that, at this time, Haplosporidium sp. is not responsible for these events in Campanian and Sicilian waters.

Epidemiology and clinical features of Mediterranean spotted fever in Italy.

Mediterranean Spotted Fever is caused by Rickettsia conorii and is transmitted to humans by Rhipicephalus sanguineus, the common dog tick. It is characterized by the symptomatologic triad: fever, exanthema and "tache noire", the typical eschar at the site of the tick bite. In Italy the most affected region is Sicily. The seasonal peak of the disease (from June through September) occurs during maximal activity of immature stage ticks. Severe forms of the disease have been reported in 6% of patients, especially adults with one of the following conditions: diabetes, cardiac disease, chronic alcoholism, glucose-6-phosphate dehydrogenase deficiency, end stage kidney disease. The mortality rate may reach 2.5%. Oral or parenteral administration of tetracyclines or chloramphenicol represent the standard treatment. Recent studies indicate that oral clarithromycin and azithromycin could constitute an acceptable alternative for the treatment of the disease in children; furthermore, they could be recommended during pregnancy.

Use of sulesomab in the diagnosis of brucellar spondylitis.

Twenty-two patients with suspected brucellar spondylitis were investigated to evaluate the possible diagnostic role of Sulesomab, a (99m)Tc-antigranulocyte antibody Fab' fragment. Sensitivity and specificity were compared with those of magnetic resonance imaging (MRI). Skeletal involvement was detected by MRI in 11 cases, while leukoscintigraphy indicated normal vertebral uptake in seven of these patients, increased uptake in two patients, and decreased uptake in two patients. Leukoscintigraphy of the 11 patients negative by MRI demonstrated increased uptake in two cases. The sensitivity and specificity of leukoscintigraphy were 27.2% and 81.1%, respectively. Based on these results, leukoscintigraphy is not indicated for the management of patients with suspected brucellar spondylitis.

Anxiety and high plasma catecholamines do not impair pharmaco-induced erection of psychogenic erectile dysfunctional patients.

The aim of this study was to assess the influence of anxiety and plasma catecholamines on the pharmaco-induced erection of psychogenic erectile dysfunction (ED) patients. A total of 23 patients with psychogenic ED aged from 19 to 43 y were submitted to: (1) anxiety evaluation by the Spielberger's State and Trait Anxiety Inventory-STAI; (2) intracavernous injection of PGE1 10 microg+phentolamine 1 mg with the response monitored by Rigiscan; (3) blood sampling from cavernous bodies and cubital vein for adrenaline and noradrenaline levels determination by high performance liquid chromatography. The whole procedure was done in a single clinical setting at the same day. We found no significant correlation between the erection rigidity and the cavernous or peripheral catecholamines or between erection rigidity and anxiety scores. Some patients showed rigid erections despite high anxiety scores or penile catecholamine levels while others, with incomplete erections, had much smaller levels. These results are suggestive of a more complex mechanism controlling the penile sympathetic responsiveness in psychogenic ED patients.

Effect of chemical stimulation of the dorsomedial hypothalamic nucleus on blood plasma glucose, triglycerides and free fatty acids in rats.

The effects of chemical stimulation of the dorsomedial hypothalamic nucleus (DMH) on blood plasma concentration of glucose, triglycerides, insulin, and free fatty acids (FFA) were investigated in anesthetized adult Wistar rats. Microinjection of 12.5 nmol of norepinephrine into the DMH increased blood plasma concentration of glucose and FFA, decreased triglycerides, and did not change plasma insulin within 5 min; after 20 min, blood glucose and FFA reached control values. Microinjection of epinephrine (12.5 nmol) into the DMH also increased blood plasma glucose concentration and decreased triglycerides after 5 min. These effects are probably mediated by beta-adrenergic mechanisms, because they were prevented by beta-adrenergic antagonist propranolol, but not by alpha-adrenergic antagonist prazosin. Microinjection into the DMH of glutamate, dopamine, or acetylcholine failed to cause any change in those metabolic parameters, corroborating the hypothesis that the DMH is part of a beta-adrenergic pathway involved in short-term modulation of the availability of glucose and FFA.

Increase of food intake induced by glucagon in the rat.

The effect of intraperitoneal administration of saline, glucose (25 mg/100 g b.w.), insulin (0.025 U/100 g b.w.) and glucagon (50 micrograms/kg b.w.) on glycemia, liver glycogen concentration and food intake was studied on 104 male adult Wistar rats. When saline was injected the amount of food ingested was similar to that expected at the metabolic moment selected for the tests. Glucose administration did not reduce food intake but both insulin and glucagon provoked a threefold increase during the 60 minutes ensuing the injection. The overall ingestion of food during the 24 hours after the injection of the hormones was significantly higher (about 10%) than the control values during the preceding or the succeeding 24 hours. A hyperphagic, rather than a hypophagic effect of glucagon administration is possibly related to the small dose used in the experiments. The mechanisms involved in the increase of food intake due to glucagon are discussed in terms of acceleration of the metabolic reactions that normally prevent large drops of glycemia as glucose utilization proceeds during the inter-meal periods and that in physiological conditions build up until the need for food arises.

Meal-feeding and physical effort. 1. Metabolic changes induced by exercise training.

To evaluate the consequences of the combination of meal-feeding (which causes in the long term several adaptations that lead to saving stored energetic substrates), rats subjected to a 2-hr feeding/22-hr fasting schedule were forced to swim 30 min everyday at a fixed hour during four weeks. The results indicate that meal-fed exercised rats: 1) increase food intake above that found in the nonexercising and the corresponding (nonfood-restricted) controls; 2) did not lose weight (in contrast to the controls); 3) initially had a high glycogen mobilization but at the end of the fourth week started to save hepatic glycogen again, despite the intense energy demanding exercise; 4) maintained a slight hyperglycemia; 5) mobilized less free fatty acids than the nonexercising meal-fed rats, probably due to higher insulinemia; 6) had a lower content of ascorbic acid in the adrenal glands in comparison to the control exercising rats; this suggests that the exercise was less stressful in the latter group.

Meal-feeding and physical effort. 2. Metabolic changes induced by an acute exercise.

In this study, rats under meal-feeding up to four weeks were submitted to a sudden exercise (swimming) to evaluate the effects of a behavior that requires mobilization of a large amount of energy on some physiological parameters already changed due to the chronic food-restriction. During exercise meal-fed rats: 1) increase the rate of gastric emptying; 2) maintain glycemia more steadily than the controls even during a long-lasting exercise; 3) maintain high liver glycogen concentration and its mobilization starts later on; 4) free fatty acid mobilization is lower than in the controls but during exercise do use much more; 5) keep more glycogen in the muscles (including the heart) than the controls but during the exercise do utilize much more; 6) are slightly less stressful (mainly after a longer exercise) than the controls as suggested by the adrenal ascorbic acid content.

Carbohydrate metabolism and food intake in food-restricted rats. Relationship between the metabolic events during the meal and the degree of food intake.

To study some metabolic features during feeding in food-restricted rats two groups of animals were maintained on a 2 hr feeding/22 hr fast schedule. Group D (n = 38) received a meal every day from 8:00 to 10:00 a.m. Group N (n = 34) was given the meal from 8:00 to 10:00 p.m. The average total amount of food ingested by rats of group N in the two hour period was 6.3 +/- 0.4 whereas Group D ingested 4.8 +/- 0.3 g/100 g b.w. The metabolic pattern also was different in one group as to the other. The basal liver glycogen content when feeding started was considerably lower in the nocturnal group (0.14 +/- 0.02 mg/100 mg of liver tissue) than in the diurnal group (0.44 +/- 0.10 mg/100 mg). Afterwards glycogen increased in both groups but more steeply and intensely in group N. Glycemia increased in group D and was almost invariant in group N. Insulinemia went up in both groups but in group D its peak was higher and occurred 60 minutes after the onset of feeding whereas the peak in group N was much lower and occurred at 90 minutes. There was a clear dissociation between the time courses of insulinemia and glycemia in both groups, especially in group N, which suggests a central control of insulin secretion during feeding that partially unlocks it from blood glucose concentration. The hepatic glycogen content was partially linked to the amount of food ingested but again there was a dissociation between these two variables, inasmuch as a higher glycogen replenishment in the nocturnal group corresponded to a larger food intake.

Reversibility of metabolic changes induced by feeding schedule in rats.

This study aimed to investigate the effect of free-feeding on rats kept on meal-feeding schedule for a prolonged period. Thus, rats meal-fed for 4 and 20 weeks were given free access to food for subsequent 5 weeks. The metabolic adaptation of higher hepatic glycogen content, low plasma FFA values and sustained glycemia during 22-hr fast, reported for rats subjected to meal-feeding, completely disappeared after free-eating period. The rate of body weight gain increased as a consequence of the free access to food in both groups but the control values (group with food ad lib all the time) were attained only in rats previously submitted to meal-feeding for the shorter period of time (4 weeks). The findings of this study suggest that the recovery of body weight by meal-fed rats, for the control values, seems to depend on the duration of the meal-feeding schedule and the age when it is imposed.