Humoral immunity response to human endogenous retroviruses K/W differentiates between amyotrophic lateral sclerosis and other neurological diseases.

BACKGROUND AND PURPOSE: Human endogenous retroviruses (HERV) K/W seem to play a role in fostering and exacerbation of some neurological diseases, including amyotrophic lateral sclerosis (ALS). Given these findings, the immunity response against HERV-K and HERV-W envelope surface (env-su) glycoprotein antigens in serum and cerebrospinal fluid (CSF) was investigated for ALS, multiple sclerosis (MS) and Alzheimer's disease patients and in healthy controls. METHODS: Four antigenic peptides derived respectively from HERV-K and HERV-W env-su proteins were studied in 21 definite or probable ALS patients, 26 possible or definite relapsing-remitting MS patients, 18 patients with Alzheimer's disease and 39 healthy controls. An indirect enzyme-linked immunosorbent assay was set up to detect specific antibodies (Abs) against env-su peptides. RESULTS: Amongst the measured levels of Abs against the four different HERV-K peptide fragments, only HERV-K env-su19-37 was significantly elevated in ALS compared to other groups, both in serum and CSF. Instead, amongst the Abs levels directed against the four different HERV-W peptide fragments, only HERV-W env-su93-108 and HERV-W env-su248-262 were significantly elevated, in the serum and CSF of the MS group compared to other groups. In ALS patients, the HERV-K env-su19-37 Abs levels were significantly correlated with clinical measures of disease severity, both in serum and CSF. CONCLUSIONS: Increased circulating levels of Abs directed against the HERV-W env-su93-108 and HERV-W env-su248-262 peptide fragments could serve as possible biomarkers in patients with MS. Similarly, increased circulating levels of Abs directed against the HERV-K env-su19-37 peptide fragment could serve as a possible early novel biomarker in patients with ALS.

Prevalence of foot disorders in captive Sardinian partridges (Alectoris barbara barbara Bonnaterre, 1790) as useful indicators of fitness to natural environment.

Foot health represents an issue in wild avifauna breeding practices. In particular, prevalence of digital ulceration (DU) and foot pad dermatitis (FPD) are valid indicators of welfare in wildlife conservation centres and may be interpreted as to fitness for bird's reintroduction into the wild. This study meant to test the effects of raising practices on foot pad health in captive Sardinian partridges (Alectoris barbara barbara Bonnaterre, 1790) reared for biodiversity conservation, to assess welfare and fitness to reintroduction into nature. A total of 22 couples were allotted into two experimental groups. In one group, 10 couples were housed in 10 cages for breeding partridges, consisting of two animals each, with metal wire flooring system, above trays where droppings were collected. The remaining 12 couples were housed in six aviaries, consisting of four animals each, on natural (earth and stones) ground. In both groups, partridges were fed identical diets. No significant differences of food pad scoring were found between birds housed in cages (2.3 ± 0.4) and those reared in aviaries on natural ground (2.5 ± 0.6). Moreover, scores of male foot pads for both groups (2.4 ± 0.6) had no significant differences in comparison with female foot pads, independently on housing (2.5 ± 0.4). Body mass (BM) was higher (+4.36%) than average BM reported for wild Sardinian partridges. Digital ulceration was found in the 20% of females, exclusively from the cage group. Body mass of females in cages with metal wire flooring appeared to be significantly (p < .001) and negatively correlated (r = -.528) with DU prevalence. These results suggest that housing conditions impacts differently on behaviour of females and males in one same couple, and this relates to foot health, in particular as to DU prevalence.

Morphometric traits of gizzard in relation to feeding habits of wild Sardinian partridges (Alectoris barbara barbara, Bonnaterre, 1790) with particular regard to clast selection.

Housing and feeding practices of wild birds for conservation management of biodiversity or restocking play a crucial role in determining the survival rates of animals when released into nature. Failure in coping with the environment might be one of the main flaws captive animals can experience when put into natural habitat. The present investigation aimed at exploring feeding habits and related morphometric traits of gizzard with respective content from wild partridges in comparison with captive ones. A total of 52 hunted wild Sardinian adult partridges (Alectoris barbara barbara Bonnaterre, 1790) were used. By comparison, 42 captive adult partridges reared in cages were enrolled. From each animal, the morphology of gizzard was investigated and respective content analysed for gross composition and taxonomical determination of fractions. Wet sieving analysis of each gizzard content was carried out (four-sieve towers with different mesh sizes: 1 mm, 500 µm, 250 µm and 125 µm), and relative and absolute weight of fresh filled and empty gizzards were recorded. Thickness of muscular layer of gizzard wall was measured by stereomicroscope. Carcass weight significantly (p < 0.05) differed between captive vs. wild partridges (478 ± 21 and 305 ± 35 g respectively). Post-mortem inspection highlighted gross morphological differences of gizzards between the two groups. Fresh weight of empty gizzards was 6.37 ± 0.80 vs. 11.25 ± 1.82 g, with average pH values of digesta 4.97 ± 0.11 vs. 4.38 ± 0.28 in captive vs. wild partridges respectively. Gizzard content from wild partridges accounted a 61.7% vs. 38.3% of biological vs. non-biological material proportions (DM basis). The non-biological material was mostly represented by lithic fragments and minerals (quartz, feldspar, calcite and mica) with specific peculiarities in terms of granulometry and morphometry. Feeding the captive partridges should point to support morphological and functional adaptation of gizzards to the feeding stuffs naturally available in the environment.

Multiple sclerosis-associated retrovirus and progressive disability of multiple sclerosis.

Retrovirus-like particles containing the multiple sclerosis-associated retrovirus RNA, significantly found in the cerebrospinal fluid of patients with multiple sclerosis, have been preliminarily associated with a short-term poor clinical and radiological prognosis of the disease. We asked whether these prognostic indications are still measurable after a long-term clinical evaluation (10 years). Our 10-year blind observational study confirms that the presence of multiple sclerosis-associated retrovirus in the cerebrospinal fluid of early multiple sclerosis patients is associated with a significantly greater rate of relapse-unrelated unremitting disability and secondary progression of the disease.

Homologous HSV1 and alpha-synuclein peptides stimulate a T cell response in Parkinson's disease.

Environmental factors are implicated in the development of Parkinson's disease (PD). The aim of this study is to investigate the role of cell-mediated immunity upon a specific immune-stimulation with HSV-1 and human alpha-synuclein homologues peptides by using the intracellular cytokine method on Parkinson's patients and healthy controls. The study showed, for the first time, a specific response to TNF-α CD8, CD4 and NK cells after stimulation in PD patients. Our data show a possible role of the immune system in the pathogenesis of Parkinson's disease, and that HSV-1 infections may lead to a progression of the disease.

Multiple sclerosis-associated retrovirus and MS prognosis: an observational study.

MS-associated retrovirus (MSRV) in the CSF may have gliotoxic properties and could be associated with a more disabling MS. The authors tested this hypothesis in 15 untreated patients with MS: 6 MSRV- and 9 MSRV+ at the time of CSF withdrawal. After a 3-year mean follow-up, MSRV- patients showed a stable MS course, whereas MSRV+ patients had a progressive course (p = 0.01).

[Toxicity of atrazine and its metabolite deethylatrazine in Thamnocephalus platyurus and Dugesia gonocephala]. / Tossicità dell'atrazina e del suo metabolita desetilatrazina per il Thamnocephalus platyurus e per la Dugesia gonocephala.

The toxicity of Atrazine and that of its Desethylatrazine metabolite has been defined employing two organisms already tested in our labs the Dugesia gonocephala, belonging to a scissiparous strain coming from the island of Tavolara (Sardinia) and the Thamnocephalus platyurus, crustacean anostracan produced under form of quiescent cysts from Creasel Ltd. (Deinze, Belgium). It has been defined the lethal concentrations at 50% of Atrazine and of Desethylatrazine, of which it has been studied also the report dose-effect in the comparisons of the rectilinear motility. The results highlight that in the comparisons of the T. platyurus Atrazine expounds a toxicity of around three times that of its metabolite; this is not seen with the Planarians that are equally sensitive to both the compounds. The use of these two experimental models for the evaluation of the contamination of bodies of water is shown to be particularly useful given their great sensitivity to pollutants.

[Eco-toxicological study conducted with a battery of biological and phytological tests on sediments carried out on a series of 24 tributaries of the Po in 1994 and 1995]. / Ricerche ecotossicologiche eseguite con batterie di tests biologici e fitologici sui sedimenti prelevati in sequenza da 24 fiumi affluenti del Po, negli anni 1994 e 1995.

In spring and autumn 1994 and 1995 affluent water and bed sediment were sampled from 24 tributaries of the Po river, always at the same site and at the nearest place to the confluence. In the laboratory the pore water was separated from the particle fraction of the sediment. The organic compounds bound to the latter component were extracted with solvents and brought to water solution by means of dimethyl sulfoxide. The observed animal species, along with the seeds of Lepidium, were exposed to effluent water, to pore water and to water solutions of the organic compounds extracted from bed sediment. Toxicity was evaluated on the basis of 1) direct lethality, 2) the delay of embryo development, 3) the impairment of regeneration, in the animal species, while the germination index was used for the Lepidium susceptibility. The results of these investigations demonstrate that 1) the challenged species cover a broad range of sensitivities toward environmental toxins, 2) toxicity found in river samples appears almost exclusively bound to the sediment, 3) the noxious effects found in the tributaries of the Po river increase moving downstream, and 4) likewise the sediment-bound toxicity varies among the different samplings, both as a consequence of changes in rain-dependent river flow, and because of the man-made interventions on the river sides and bed.

Multiple sclerosis: reduced proportion of circulating plasmacytoid dendritic cells expressing BDCA-2 and BDCA-4 and reduced production of IL-6 and IL-10 in response to herpes simplex virus type 1.

OBJECTIVE: We hypothesized that autoaggressive immune responses observed in multiple sclerosis (MS) could be associated with an imbalance in proportion of immune cell subsets and in cytokine production in response to infection, including viruses. METHODS: We collected blood mononuclear cells (MNC) from 23 patients with MS and 23 sex- and age-matched healthy controls (HC) from the island of Sardinia, Italy, where the prevalence of MS is extraordinarily high. Using flow cytometry, we studied MNC for expression of blood dendritic cell antigens (BDCA)-2 and BDCA-4 surface markers reflecting the proportion of plasmacytoid dendritic cells (pDC) that produce type I interferons (IFNs) after virus challenge and promote Th2/anti-inflammtory cytokine production. In parallel, pro-inflammatory (interleukin [IL]-2, IL-12, IFN-gamma), anti-inflammatory (IL-4, IL-10), and immuno-regulatory/pleiotropic cytokines (type I IFNs including IFN-alpha and beta, IL-6) were measured before and after an in vitro exposure to herpes simplex virus type 1 (HSV-1). RESULTS: The subset of lineage negative (lin(-)), BDCA-2(+) cells was lower in patients with MS compared with HC (0.08 + or - 0.02% vs 0.24 + or - 0.02%; P < 0.001). A similar pattern was observed for lin(-)BDCA-4(+) cells (0.08 + or - 0.02% vs 0.17% + or - 0.03; P < 0.01). Spontaneous productions of IL-6 (45 + or - 10 pg/mL vs 140 + or - 26 pg/mL; P < 0.01) and IL-10 (17 + or - 0.4 pg/mL vs 21 + or - 1 pg/mL; P < 0.05) by MNC were lower in patients with MS compared with HC. Spontaneous production of IL-6 (6.5 + or - 0.15 pg/mL vs 21 + or - 5 pg/mL; P < 0.01 and IL-10 (11 + or - 1 pg/mL vs 14 + or - 3 pg/mL; P < 0.05) by pDC was also lower in patients with MS compared with HC. Exposure of MNC to HSV-1 showed, in both patients with MS and HC, increased production of IFN-alpha, IL-6, and IL-10 but decreased production of IL-4. In response to HSV-1 exposure, productions of IL-6 (165 +or - 28 pg/mL vs 325 + or - 35 pg/mL; P < 0.01) and IL-10 (27 +or - 3 vs 33 + or - 3 P < 0.05) by MNC as well as by pDC (IL-6: 28 + or - 7 vs 39 + or - 12 P < 0.05; IL-10: 14 + or - 1 vs 16 + or - 3 P < 0.05) were lower in patients with MS compared with HC. CONCLUSION: The results implicate a new evidence for altered immune cells and reduced immune responses in response to viral challenge in MS.

Multiple sclerosis-associated retrovirus and optic neuritis.

One prognostic factor for early multiple sclerosis (MS) patients to develop a definite MS may be the presence of the MS-associated retrovirus (MSRV) in the cerebrospinal fluid (CSF). We designed a specific study on a cohort of optic neuritis (ON) patients to evaluate the MSRV-dependent conversion to MS relative to the prediction conferred by magnetic resonance imaging (MRI) and CSF abnormalities. At follow-up, 33.3% MSRV+ and 0% MSRV- ON patients developed MS (P = 0.03). The prediction value is lower than that given by CSF and MRI abnormalities (42.3%). This intriguing finding is discussed in the light of the abundant discrepancies observed in the MSRV literature. Multiple Sclerosis 2006; 12: 357-359. www.multiplesclerosisjournal.com

Multiple sclerosis-associated retrovirus in early multiple sclerosis: a six-year follow-up of a Sardinian cohort.

The human endogenous retroviruses (HERV)-W family contains an extracellular particle detected in multiple sclerosis (MS) patients and designated as MS-associated retrovirus (MSRV). Through nested RT-PCR assays specific for pol MSRV gene, we preliminary reported that its presence in the cerebrospinal fluid (CSF) of early MS patients could be indicative of a poor prognosis upon a three-year follow-up. In the present clinical study, we enlarged our blind observation up to six years. At study entry, 10 MS patients were MSRV+ and eight were MSRV- in the CSF, both groups having a similar mean age and Expanded Disability Status Scale (EDSS) score. After six year follow-up, the mean EDSS significantly differed between the MSRV+ and MSRV- cohorts (4.3 versus 2.2; P = 0.004), as did the annual relapse rate (0.5 in the MSRV+ versus 0.3 in the MSRV-; P = 0.01). Finally, two MSRV+ patients entered the progressive phase, whilst none of the MSRV- group entered this phase, and 9/10 MSRV+ versus 2/8 MSRV patients were treated with immunomodulatory or immunosuppressive drugs (P = 0.009). In conclusion, we found that the presence of MSRV virions in the CSF at the onset of MS is associated, not only with disability accumulation, but also with a higher rate of clinical re-exacerbations. With the known potential pathogenic effects of MSRV given in the literature, further investigations on MSRV are warranted.

Immunomodulation of fucosyl-lactose and lacto-N-fucopentaose on mononuclear cells from multiple sclerosis and healthy subjects.

The 1,2-fucosyl-oligosaccharides, and among these the 2'-fucosyl-lactose (2'-FL) and lacto-N-fucopentaose (LNFP)-I, are quantitatively the most represented oligosaccharides of human milk. They are also seen to represent an important immune device to prevent nursing infants from severe infectious diarrhoea. Recent evidences show that the appearance of 2'-FL and LNFP-I in human colostrums is synchronised with the macrophage inhibition and that LNFP-III induces a Th2 response from the mouse peripheral immune system. Since mannosyl-fucosyl receptors are described on the macrophage surface, all these evidences allow us to investigate on the possible immune function of human 2'-FL and LNFP-I in vitro on LPS-activated mononuclear cells (MNC) from 12 patients with multiple sclerosis (MS) and 20 matched health controls (HC). We found that 2'-FL and LNFP-I significantly decrease, to a different extent, the MNC proliferation from both HC and MS patients, in a linear and dose-dependent manner. 2'-FL and LNFP-I also reduce the production of IL-12 and IFN-γ, particularly in MS patients as compared to HC (p=0.01 and p<0.001, respectively), while increasing that of IL-10. The overall immunomodulatory effect of 2'-FL and LNFP I here presented may represent a future therapeutic option for the abnormal immune response found in some monocyte-mediated diseases.

Glatiramer acetate reduces lymphocyte proliferation and enhances IL-5 and IL-13 production through modulation of monocyte-derived dendritic cells in multiple sclerosis.

Dendritic cells (DC), as the most effective antigen presenting cells, are protagonists of the complex immune network involved in multiple sclerosis (MS) lesion formation. Glatiramer acetate (GA), a synthetic random copolymer, is thought to exert its therapeutical effect in MS by favouring both Th2 cell development and IL-10 production from peripheral lymphocytes as well as by systemically affecting the antigen presenting cells. In the present study we further analysed the mechanisms of action of GA by using an autologous DC-lymphocytes (Ly) coculture system from 11 MS patients and 12 matched healthy controls (HC). We found that, in MS patients, pretreatment with GA significantly decreases the in vitro proliferative effect of DC on lymphocytes as compared to HC and to unpulsed or myelin basic protein (MBP)-pulsed DC from MS patients (P < 0.05). In addition, GA-treated DC from both MS patients and HC significantly increase the lymphocyte production of IL-5 and IL-13 as compared to MBP-treated DC (P < 0.05). In conclusion our in vitro study may provide new therapeutical mechanisms of GA on lymphocytes, antiproliferative and Th2-favouring effects, which are mediated by monocyte-derived DC.

Intrathecal chitotriosidase and the outcome of multiple sclerosis.

Activated macrophages are major effectors at all stages of lesion formation in multiple sclerosis (MS) brain. Here, we report that the macrophage enzyme chitotriosidase (Chit) is significantly elevated both in plasma and cerebrospinal fluid (CSF) of patients with MS as compared to healthy controls and other neurological patients (P<0.001). Furthermore, the Chit activity in blood significantly associates with the MS clinical course (higher in secondary progressive relative to relapsing-remitting, P=0.01) and the clinical severity as measured by Kurtkze's Expanded Disability Status Scale (P<0.001). Also, we found that Chit activity is compartmentalized in the central nervous system of early MS patients and that its CSF/plasma quotient, in the presence of a preserved albumin quotient, correlates with the extent of future clinical deterioration (r=0.91; P<0.001). These findings confirm that innate immunity, here represented by Chit, is clinically relevant in MS and allows, if confirmed, reconsidering novel MS therapeutic strategies specifically aimed at this branch of the immune response.

Inflammatory biomarkers in blood of patients with acute brain ischemia.

Although many failed surrogate markers are provided in the literature, inflammation may contribute to the outcome of ischemic stroke. In 50 consecutive patients with acute ischemic stroke, in the absence of symptoms and signs of concomitant infection, we evaluated a panel of biomarkers reported to be variably associated with brain ischemia, and correlate their serum level with the brain lesion volume and clinical outcome. Infarct size was calculated on computed tomography (CT) scans by means of the Cavalieri's method. Neurological impairment was scored by using the Glasgow Coma Scale, Glasgow Outcome Scale and National Institutes of Health (NIH) scales at stroke onset and 3-month follow-up. Some markers showed a direct significant correlation with both initial and final NIH scale and with infarct size, particularly tumor necrosis factor alpha (TNF-alpha) (P=0.002), intercellular adhesion molecule-1 (P<0.01) and matrix metalloproteinase-2/9 (P=0.001). In contrast to previous reports, interleukin-6 (IL-6) serum level showed a significant inverse correlation with both final neurological impairment and infarct size (P<0.001). This novel finding allows us suggesting that IL-6, in the context of a complex pro-inflammatory network occurring during stroke, is associated with neuroprotection rather than neurotoxicity in patients with ischemic brain injury.

[Toxicologic effects of cadmium on gametes and embryonal kinetics of paracentrotus lividus]. / Effetti tossicologici da cadmio sui gameti e sulla cinetica embrionale del paracentrotus lividus.

The adverse effects of cadmium chloride or cadmium sulphate on both fertilization of sea urchin eggs by spermatozoa and cadmium treated embryos have been studied. Cd treated ova can be fertilized by control spermatozoa. On the contrary, the preliminary treatment of spermatozoa with CdCl2 or CdSO4 succeeds in reducing the fertilization rate of control eggs at concentrations as low as 2 micrograms/ml. Cd sulphate appears to be more noxious than Cd chloride. The study of the residual motility of sea urchin embryos exposed to either CdCl2 or CdSO4 has evidenced a fairly effect at 21 hours after fertilization. On the contrary, concentrations of 25 micrograms/ml Cd chloride or Cd sulphate significantly affect the motility of 48 hours embryos. Higher levels of cadmium, for example, 150 or 200 micrograms/ml, completely block the translation of developing organisms. In this instance, Cd chloride is more effective than Cd sulphate. In addition, a few embryos are affected by malformations and are underdeveloped. These results are interpreted on the basis of the inhibitory effect of cadmium on the CaCO3 uptake by tissues and structures which physiologically need to be calcified in order to develop and function. Further, they give support to the predictive significance of the toxicological tests on the sea urchin plutei in monitoring environmental hazards.

[Embryotoxicity of fenbendazole in Paracentrotus lividus]. / Embriotossicità del fenbendazolo in Paracentrotus lividus.

The aim of the present work was to evaluate the embryotoxicity of Fenbendazole, a benzimidazole carbamate-derived anthelmintic drug widely employed in Veterinary Medicine, by using the embryonal development of Paracentrotus lividus (sea urchin) as a experimental model. Embryos were obtained by in vitro eggs fertilization and cultured in seawater. Five embryo suspensions were added by Fenbendazole reaching a final concentration of 5 micrograms/l, 7.5 micrograms/l, 10 micrograms/l, 12.5 micrograms/l and 25 micrograms/l; a suspension was kept drug-free as a control. Embryo development was evaluated by microscopical examination of suspensions at 3 and 40 hours. Our results show that a concentration of 5 micrograms/l of the drug determines a considerable delay of the embryonal development in the 95 percent of the elements observed, and a concentration of 25 micrograms/l produces a block of the embryogenesis at the phase of morula and blastula in all embryos. Results confirm that the effects observed are probably due to an extended inhibition of several enzyme complexes of the embryo cells.

Serum uric acid and multiple sclerosis.

Several studies indicate that patients with multiple sclerosis (MS) have low serum levels of the endogenous antioxidant uric acid (UA), although it has not been established whether UA is primarily deficient or secondarily reduced due to its peroxynitrite scavenging activity. We measured serum urate levels in 124 MS patients and 124 age- and sex-matched controls with other neurological diseases. In addition, we compared UA levels when MS patients were stratified according to disease activity (by means of clinical examination and MRI), duration, disability and course. MS patients had significantly lower serum urate levels than controls (p= 0.001). However, UA levels did not significantly correlate with disease activity, duration, disability or course. Our study favors the view that reduced UA in MS is a primary, constitutive loss of protection against oxidative agents, which deserves further pathogenetic elucidation aimed at future therapeutic strategies.