Search

Glial cells in the gut represent the morphological and functional equivalent of astrocytes and microglia in the central nervous system (CNS). In recent years, the role of enteric glial cells (EGCs) has extended from that of simple nutritive support for enteric neurons to that of being pivotal participants in the regulation of inflammatory events in the gut. Similar to the CNS astrocytes, the EGCs physiologically express the S100B protein that exerts either trophic or toxic effects depending on its concentration in the extracellular milieu. In the CNS, S100B overexpression is responsible for the initiation of a gliotic reaction by the release of pro-inflammatory mediators, which may have a deleterious effect on neighboring cells. S100B-mediated pro-inflammatory effects are not limited to the brain: S100B overexpression is associated with the onset and maintenance of inflammation in the human gut too. In this review we describe the major features of EGCs and S100B protein occurring in intestinal inflammation deriving from such.

Enteric glial cells (EGC) actively mediate acute and chronic inflammation in the gut; EGC proliferate and release neurotrophins, growth factors, and pro-inflammatory cytokines which, in turn, may amplify the immune response, representing a very important link between the nervous and immune systems in the intestine. Cannabidiol (CBD) is an interesting compound because of its ability to control reactive gliosis in the CNS, without any unwanted psychotropic effects. Therefore the rationale of our study was to investigate the effect of CBD on intestinal biopsies from patients with ulcerative colitis (UC) and from intestinal segments of mice with LPS-induced intestinal inflammation. CBD markedly counteracted reactive enteric gliosis in LPS-mice trough the massive reduction of astroglial signalling neurotrophin S100B. Histological, biochemical and immunohistochemical data demonstrated that S100B decrease was associated with a considerable decrease in mast cell and macrophages in the intestine of LPS-treated mice after CBD treatment. Moreover the treatment of LPS-mice with CBD reduced TNF-α expression and the presence of cleaved caspase-3. Similar results were obtained in ex vivo cultured human derived colonic biopsies. In biopsies of UC patients, both during active inflammation and in remission stimulated with LPS+INF-γ, an increased glial cell activation and intestinal damage were evidenced. CBD reduced the expression of S100B and iNOS proteins in the human biopsies confirming its well documented effect in septic mice. The activity of CBD is, at least partly, mediated via the selective PPAR-gamma receptor pathway. CBD targets enteric reactive gliosis, counteracts the inflammatory environment induced by LPS in mice and in human colonic cultures derived from UC patients. These actions lead to a reduction of intestinal damage mediated by PPARgamma receptor pathway. Our results therefore indicate that CBD indeed unravels a new therapeutic strategy to treat inflammatory bowel diseases.

Glioneuronal tumors are an increasingly recognized cause of partial seizures that occur primarily in children and young adults. Focal epilepsy associated with glioneuronal tumors is often resistant to pharmacological treatment. The cellular mechanisms underlying the epileptogenicity of glioneuronal tumors remain largely unknown. The involved mechanisms are certain to be multifactorial and depend on specific tumor histology, integrity of the blood-brain barrier, characteristics of the peritumoral environment, circuit abnormalities, or cellular and molecular defects. Glioneuronal tumors presenting with epilepsy were observed to have relatively benign biological behavior. The completeness of the tumor resection is of paramount importance in avoiding tumor progression and malignant transformation, which are rare in cases of epileptogenic glioneuronal tumors. An evolving understanding of the various mechanisms of tumor-related epileptogenicity may also lead to a more defined surgical objective and effective therapeutic strategies, including antiepileptogenic treatments, to prevent epilepsy in at-risk patients.

In the central nervous system glial-derived S100B protein has been associated with inflammation via nitric oxide (NO) production. As the role of enteroglial cells in inflammatory bowel disease has been poorly investigated in humans, we evaluated the association of S100B and NO production in ulcerative colitis (UC). S100B mRNA and protein expression, inducible NO synthase (iNOS) expression, and NO production were evaluated in rectal biopsies from 30 controls and 35 UC patients. To verify the correlation between S100B and NO production, biopsies were exposed to S100B, in the presence or absence of specific receptor for advanced glycation end-products (RAGE) blocking antibody, to measure iNOS expression and nitrite production. S100B and iNOS expression were evaluated after incubation of biopsies with lipopolysaccharides (LPS) + interferon-gamma (IFN-gamma) in the presence of anti-RAGE or anti-S100B antibodies or budesonide. S100B mRNA and protein expression, iNOS expression and NO production were significantly higher in the rectal mucosa of patients compared to that of controls. Exogenous S100B induced a significant increase in both iNOS expression and NO production in controls and UC patients; this increase was inhibited by specific anti-RAGE blocking antibody. Incubation with LPS + IFN-gamma induced a significant increase in S100B mRNA and protein expression, together with increased iNOS expression and NO production. LPS + IFN-gamma-induced S100B up-regulation was not affected by budesonide, while iNOS expression and NO production were significantly inhibited by both specific anti-RAGE and anti-S100B blocking antibodies. Enteroglial-derived S100B up-regulation in UC participates in NO production, involving RAGE in a steroid insensitive pathway.

We measured parallel upper critical magnetic fields in Nb/PdNi and Nb/CuNi bilayers and Nb/PdNi/Nb and Nb/CuNi/Nb trilayers. In the bilayers case the measurements reveal a dependence of the reduced two-dimensional-three-dimensional crossover temperature, t(cr) = T(cr)/T(c), on the different values of the superconductor (S)/ferromagnet (F) interface transparencies. In the case of the trilayers we observe that the reduced crossover temperature as a function of the ferromagnetic layer thickness, d(F), reaches a value equal to one only when the π-phase occurs in these systems. Also in this case the influence of the interface transparency on the observed results is discussed.

We study the vortex dynamics in the instability regime induced by high dissipative states well above the critical current in Nb superconducting strips. The magnetic field and temperature behavior of the critical vortex velocity corresponding to the observed dynamic instability is ascribed to intrinsic non-equilibrium phenomena. The Larkin-Ovchinnikov (LO) theory of electronic instability in high velocity vortex motion has been applied to interpret the temperature dependence of the critical vortex velocity. The magnetic field dependence of the vortex critical velocity shows new features in the low-field regime not predicted by LO.

We describe a single-center pediatric experience with 1 mg/kg/wk cidofovir without probenecid in 7 children with BK virus-associated hemorrhagic cystitis. Clinical improvement was observed in all cases, without adverse events, although significant reduction of urinary viral load was observed 2 weeks after the end of cidofovir in 5 out of 6 patients who completed the treatment.

Retroperitoneal schwannoma is a very rare benign tumour that accounts for only a small percentage of retroperitoneal tumours. The preoperative diagnosis is difficult and the only treatment is complete surgical excision. The authors report their experience with a case of a voluminous retroperitoneal mass in a 42-year- old male patient, incidentally discovered during a diagnostic search for a blunt abdominal trauma. Because of the unclear origin of this mass, the patient underwent periodic radiological and ultrasonographic examinations. Three years later, owing to a slight increase in this mass and vague abdominal discomfort reported by the patient, surgical excision was performed. Histological examination of the surgical specimen revealed an "ancient schwannoma". The patient is alive and well and has had no recurrence in the twenty months since surgery. The authors, after a short discussion of the subject, go on to examine the diagnostic and therapeutic treatments of this rare benign neoplasm, which always arouses lively clinical and scientific interest.

Mycoplasma pneumoniae (MP) is, considered to affect rarely children less than 5 yrs of age. This study was performed to describe the epidemiology and the clinical features of MP lower respiratory tract infection (LRTI) in children, presenting to a tertiary children hospital. Eleven month-longitudinal study of LRTI due to MP, diagnosed by polymerase chain reaction (PCR) on throat swab specimen, was performed. Out of 866 children with LRTI admitted to the Gaslini Pediatric Institute in Genoa, 102 had a positive PCR for MP. We found 39 preschool-aged children, 42 school-aged children and 21 young adolescent [6.20 (3.81) yrs old]. Interestingly, eight MP+ infants had <8 months of age. The commonest presentations were cough and/or fever (76.5%). Tachypnoea, upper respiratory tract involvement, diarrhoea and vomiting were more common in the <5 yr Gr as compared to the other groups. Chest X-ray was found abnormal in 76 children: consolidations were the commonest finding. Laboratory test showed that the preschool-aged children had a higher number of lymphocytes (p<0.0001) and monocytes (p=0.009). Thrombocytosis was found in 35.7% of children and was more frequent in the preschool-aged children (p=0.013). MP infection is common in preschool-aged children, including young infants, and may have different clinical presentation, as compared to older children.

Sweetened carbonated beverages are widely consumed, which has fuelled several conflicting opinions about their effects on upper digestive tract functions. We aimed to evaluate the effect of sweetened carbonated drinks, consumed with a standard meal, on gastro-oesophageal reflux, gastric emptying and gallbladder contraction and postmeal sensations in healthy subjects. Thirteen healthy volunteers (seven women, six males; median age 22 years) were tested following the intake of 300 mL sweetened water containing increasing concentrations of carbon dioxide (seven subjects), and of 300 mL sweetened commercial flavoured drink with and without carbon dioxide (six subjects). Gastro-oesophageal reflux, gastric emptying and gallbladder contraction were studied by pH-impedance, octanoic acid breath test and ultrasound respectively. Gastro-oesophageal refluxes were significantly increased 1 h after meal with both water and commercial beverages; only sweetened water without carbon dioxide determined a persistently increasing number of refluxes 2 h postmeal. No differences were found for gastric emptying, gallbladder contraction or postmeal symptoms with any of the beverages tested. This study shows that 300 mL of sweetened carbonated beverage with different levels of carbonation or a commercial soft drink do not modify the physiology of the upper digestive tract.

Viral infections are a rare complication in autologous hemopoietic stem cell transplant (HSCT) recipients but represent a frequent cause of disease after allogeneic HSCT. In the last years, there has been an increase in the number of viral diseases observed in these patients. This fact may be at least partially due to an improvement in diagnostic facilities, but the increasing number of transplant procedures and the more severe immunosuppression may also have played an important role.

Background & Aims: Enteric glia participates to the homeostasis of the gastrointestinal tract. In the central nervous system, increased expression of astroglial-derived S100B protein has been associated with the onset and maintaining of inflammation. The role of enteric glial-derived S100B protein in gastrointestinal inflammation has never been investigated in humans. In this study, we evaluated the expression of S100B and its relationship with nitric oxide production in celiac disease., Methods: Duodenal biopsy specimens from untreated and on gluten-free diet patients with celiac disease and controls were respectively processed for S100B and inducible nitric oxide synthase (iNOS) protein expression and nitrite production. To evaluate the direct involvement of S100B in the inflammation, control biopsy specimens were exposed to exogenous S100B, and iNOS protein expression and nitrite production were measured. We also tested gliadin induction of S100B-dependent inflammation in cultured biopsy specimens deriving from on gluten-free diet patients in the absence or presence of the specific S100B antibody., Results: S100B messenger RNA and protein expression, iNOS protein expression, and nitrite production were significantly increased in untreated patients but not in on gluten-free diet patients vs controls. Addition of S100B to control biopsy specimens resulted in a significant increase of iNOS protein expression and nitrite production. In celiac disease patients but not in controls biopsy specimens, gliadin challenge significantly increased S100B messenger RNA and protein expression, iNOS protein expression, and nitrite production, but these effects were completely inhibited by S100B antibody., Conclusions: Enteric glial-derived S100B is increased in the duodenum of patients with celiac disease and plays a role in nitric oxide production.

We compared growth patterns and morphology of fine roots of size-controlling and invigorating peach (Prunus persica (L.) Batsch) rootstocks. Peach trees were grafted on five rootstocks: a vigorous control (Nemaguard), three intermediate vigor rootstocks (K119-50, P30-135 and Hiawatha), and a semi-dwarfing rootstock (K146-43). Minirhizotron tubes were installed at the base of trees on each rootstock and root images captured with a minirhizotron digital camera system. Number, visible length, and diameter of new roots were recorded at fixed soil depths from April 19, 2000 to December 19, 2001. Root diameter, specific root length, root tissue density and root length density were also measured periodically for each rootstock on roots collected from in-growth cores. Rootstocks had similar seasonal patterns of new root production. Fine root production was lowest in winter and appeared to decline during the final stages of fruit growth. A rootstock with almond in its genetic background (K119-50) produced the greatest quantity of fine roots and had the greatest number of new roots below 69 cm, whereas there were no differences among the other four rootstocks in the total number of roots produced. Rootstock K146-43 had thicker fine roots than the other rootstocks. Independent of rootstock, fine roots produced during spring had greater specific root length than those produced later in the season. The seasonal pattern of fine root production did not appear to be associated with the previously reported effects of these dwarfing rootstocks on shoot growth and stem water potential early in the growing season.

S100beta is an astroglial-derived Ca2+ -binding protein having neurotrophic role on neurons and glial cells. An aberrant S100beta production has been observed in neurodegenerative disease, as Alzheimer's disease and Down syndrome. S100beta is responsible to start up a gliotic reaction by the release of pro-inflammatory mediators, including nitric oxide (NO) and cytokines from microglia and astrocytes, which are, in turn, deleterious for neurons. Interestingly, pro-inflammatory effect of S100beta seems not be restricted into the brain. Macrophages play a pivotal role in inflammatory diseases, occurring both in the brain and in the periphery. In this study, we tested the hypothesis that S100beta may affect macrophage functions, amplifying thus the inflammatory process. Our results demonstrate that S100beta stimulates both NO production and iNOS protein transcription and expression in J774 and rat peritoneal macrophages. NO production was concentration and time-dependently inhibited by two iNOS inhibitors, L-NAME and SMT. We also demonstrated that S100beta induced oxidative stress by increasing H2O2 production and lipid peroxidation of cell membrane in both macrophage types. The pro-oxidant potential of S100beta activates p38 MAP kinase (MAPK), which has been described to directly activate NF-kappaB. In our study, SB203580, a p38 MAPK inhibitor, and two NF-kappaB inhibitors, TLCK and BAY 11-7082, decreased both NO production and iNOS protein transcription and expression in S100beta-stimulated J774 and peritoneal rat macrophages. Moreover, additional studies demonstrated that S100beta affected also TNF-alpha protein expression in J774 macrophages. In conclusion, our results highlight the potential role of S100beta during an inflammatory scenario identifying macrophages as a novel S100beta-responsive cell-type.

Aim of the Study: The Authors report on a patient personally observed owing to a tumefaction, mimicking a pomelo, located on the posterior chest wall, in the same site of another past surgical operation dating back to four years ago. This tumefaction was clear at the inspection, not aching and hard-elastic at the palpation, mobile synchronously with respiratory movements, but unable to alter respiratory mechanics., Case Report: The patient was subjected to laboratory tests, which showed nothing pathological, and to instrumental tests (RX and TAC of the chest, bony scintigraphy) which showed a roundish solid tumefaction, with no "secondary" interest of bony tissue. In this case, it was executed a posterior-lateral thoracothomy, at the VI intercostal space, in the area circumscribing the past surgical scar. The careful removal of the adhesions between the mass and the costal plane, not without the sacrifice of the periosteum, permitted us the total exeresis. The anatomo-pathological test showed a desmoid fibromatosis (desmoid tumor) extra-abdominal (12.5 x 9 x 5 cm). About this kind of neoplastic masses, the risk of post-surgical relapse is very high; so many Authors consider opportune a radio-chemical adjuvant therapy. In this case, the radical excision allowed the Authors to avoid the post surgery pharmacological treatment and to get no relapses after two years from the operation.

The clinical case reported here concerns a giant lipoma (22 x 12 x 10 cm; 2740 g) located in the distal region of the right thigh, in a subfascial zone. The patient was referred to our department for a relapse of thrombophlebitis of the right lower limb (occurring about two years earlier) and presented a large tumefaction of the mid third of thigh at objective examination, the presumed onset of which dated back about ten years after a traumatic accident. Ultrasonography confirmed the presence of the tumefaction but did not allow us to identify its origin with certainty. Musculoskeletal magnetic resonance imaging, however, revealed a gross expansive lesion closely connected to the distal part of the femur, suggesting its probable benign nature. The mass was totally resected without either muscle section or the sacrifice of periosteum. The histological findings indicated an atypical lipomatous tumour. The therapy of giant lipomas is invariably surgical excision. Nevertheless, the high risk of relapse (which is typical of this kind of neoplastic proliferation) despite radical resection, makes a systematic postoperative follow-up necessary.

This study was conducted from September 2000 to September 2004 on 8 cases of inguinal canal endometriosis. All the patients presented an inguinocrural tumefaction of variable size. In 3 cases (symptomatic endometriosis), the patient's medical history yielded a preoperative diagnosis. In the other 5 cases (asymptomatic endometriosis) we obtained an incidental intraoperative diagnosis. In 2 of these, concomitant frank inguinocrural disease further hampered the preoperative diagnosis, while in the other 3 cases we were oriented towards an inguinal adeno-lymphatic disease. We found no signs of neoplastic transformation in our case series. The histological examination confirmed the diagnosis of endometriosis in all 8 cases, without any atypical cellular signs. From an analysis of the literature we deduced that the worldwide incidence of endometriosis is about 10% of the female population, though it is known that in the vast majority of cases there are few or no symptoms. Even today, despite the routine use of advanced diagnostic and surgical procedures, endometriosis foci are very often identified only incidentally owing to the greater incidence of asymptomatic or paucisymptomatic forms. The aim of our study is to stress the difficulty in diagnosing asymptomatic endometriosis and, above all, to suggest that, in the presence of a concomitant hernial pathology in a woman of child-bearing age, the diagnostic hypothesis of endometriosis should never be disregarded.

Ovarian cysts are a common pathology after the 4th decade of life. We can find either smaller functional, non-neoplastic ones (belonging to the follicular and luteinic varieties) or larger tumoral cysts, which, however, are usually benign. These may be of the serous or mucinous type and can sometimes reach really large sizes. Reports of giant ovarian manifestations were more frequent a few decades ago. Prior to the advent of modern radiological, ultrasonographic, tomographic and magnetic resonance imaging techniques, diagnosis was often difficult. Nevertheless, even today, in some cases (as a result of pronounced obesity, for example, associated perhaps with diagnostic negligence), cases of giant ovarian cysts may still be encountered. We report the case of a (previously obese) 19-year-old female, admitted to our hospital for presumed ascites, identified and ultrasonographically misdiagnosed by her gynaecologist. The patient was, in fact, suffering from giant serous cystoadenomas in both ovaries.

Fruiting kiwifruit [Actinidia deliciosa (A. Chev.) C.F. Liang et A.R. Ferguson] vines were grown in two controlled temperatures of 28 / 22 and 17 / 12°C (day / night) for 160 and 215 d, to measure shoot and fruit growth and carbon demand, and to examine competition between fruit and the shoot. Leaf area, internode lengths, fruit diameters, photosynthesis and respiration were measured at regular intervals. The net daily carbon balance per shoot was determined from the net carbon acquisition of shoots, and carbon sequestration as shoot biomass. Vines grown at high temperature had 200% more leaf area, similar stem lengths and 100% more biomass than vines grown at low temperature. Leaf area expansion and stem extension were transiently reduced when fruit growth was maximal. Photosynthetic and respiration rates were affected by temperature, leading to net carbon acquisition of 450 g shoot -1 for 28 / 22°C-grown vines and 253 g shoot -1 for 17 / 12°C-grown vines, 54% being used for leaf, stem and fruit growth. Reallocation of carbon occurred from leaves to fruit, and the consequent reduction in leaf area strongly reduced the overall carbon balance compared with vegetative vines at similar temperatures. The data support the conclusion that at low temperatures especially, there is insufficient carbon to meet the full demands of both fruit and shoot growth.

The Hartmann procedure has, in emergency colo-rectal surgery, many implications. The decision is based on clinical, radiological, instrumental and pathological findings. The authors report the results of 76 Hartmann's procedures performed between 1986 and 1995 and compare their results with those found in the current medical literature on the topic. In particular, they draw attention to an increased use of this procedures in colo-rectal emergencies; the morbility and mortality rates confirm the severity of the clinical cases that can be treated with this operation. To improve results the authors propose a therapeutic plan that uses a score for stratification of the risk (MPI, APA-CHE II, SSI, Hinchey); so the surgeon can choose the best surgical operation. Finally, the authors underline the importance of the principles of oncology surgery in colo-rectal cancer.

Several neuropathologic findings in infants and children with human immunodeficiency virus type-1 (HIV-1) infection are different from those observed in adults, probably related to the fact that the retroviral infection occurs in the setting of neuro-development. This report describes the interaction and biologic activity of tat, the HIV-1 trans-activating protein on human neuroblasts. Two human neuroblastoma cell lines, LAN-5 and GI-CA-N, have been studied for their capability to adhere to tat (full recombinant protein) and to two different peptide residues of it. Both cells adhere to tat and tat46-60 basic domain, although not to tat65-80 residue, which contains the RGD (arginine-glycine-aspartic acid) motif. Adhesion to collagen I was inhibited by preincubating GI-CA-N cells with tat,46-60 although not with tat,65-80 indicating the capability of the basic residue to interfere with collagen I-induced cellular adhesion. The expression of 200-kD neurofilaments induced by collagen I was not induced by tat,46-60 indicating that neural differentiation along the same pathway is not mimicked by this peptide. Neuroblast cell proliferation was not affected by adhesion to tat46-60 nor to tat.65-80 GI-CA-N cells are not permissive to HIV-1 infection. However, proviral DNA was documented in the cell lysate for 14 consecutive in vitro passages, whereas HIV-1 transcription was never detectable. This would exclude the possibility that tat would be transduced by these cells. GI-CA-N stained negative for CD4, although positive for Gal-C, which may explain HIV-1 entry. Results show that immature human neural cells interact with tat protein and/or its basic residue in vitro. A mechanism similar to that herein described would possibly be active in vivo, which may help in clarifying the pathogenic mechanisms of neurologic dysfunction and destruction of the CNS observed in infants infected with HIV-1.

We report an 18-year-old boy with common variable immunodeficiency who presented with splenomegaly as well as left axillary and lateral cervical lymphadenopathy. Main laboratory investigations showed severe thrombocytopenia. Epstein-Barr virus (EBV) DNA was detected in the patient's throat-washing specimens and lymph node biopsy. Lymphocytes from the lymph node biopsy were also positive for EBV nuclear antigen. Serology for EBV and cytomegalovirus was negative. A therapeutic attempt with acyclovir did not influence the course of infection. Six months' treatment with human lymphoblastoid interferon-alpha (IFN alfa) brought about the normalization of clinical and hematologic conditions. Detection on throat-washing specimens carried out 1 year after therapy was negative. Our preliminary experience suggests that human lymphoblastoid IFN-alpha is a valid alternative in therapy of immunodeficient EB virus-infected patients.

Thirty-five children with chronic HBV infection, HBV-DNA and eAg serum positivity, and HBcAg in liver tissue were treated with lymphoblastoid human interferon alpha with (16 cases) or without (19 cases) prednisolone pretreatment. The patients were double-blind randomized to receive steroid or placebo for 4 weeks, followed after 2 weeks by 5 or 10 MU/m2 interferon for 12 weeks. The e anti-e seroconversion rate reached 48%, which is much higher than the spontaneous seroconversion rate. The influence of "prednisolone priming" was not statistically significant. HBeAg clearance was similar in both groups (44% after prednisolone/interferon and 53% after interferon alone). The response to either treatment did not correlate with the pretreatment serum transaminase. HBV-DNA or degree of histological activity. Interferon was well tolerated, the side effects being less severe than in adults, and never led to suspension of the treatment.

Human immunodeficiency virus type 1 (HIV-1)-specific cytotoxic T lymphocyte (CTL) activity against different viral antigens, including env, gag/pol, and nef, was detected in peripheral blood lymphocytes of as many as 25% of uninfected children born to HIV-1-infected mothers. The relatively high frequency of this finding in uninfected, exposed children suggests that CTL-mediated immunity may play a key role in the control or even elimination of the virus. Further characterization of this natural model of potentially protective immunity could provide new insight and suggest further strategies for the development of HIV-1 vaccines.

Zidovudine (ZDV) administration during pregnancy has been suggested for the prevention of mother-to-child HIV-1 transmission. Reliable levels of the drug have been observed in the fetus and in the newborn. Seven HIV-1-infected pregnant women who declined to have abortions and whose immunological status required antiretroviral treatment were administered oral ZDV 18 mg/kg in four daily doses, the initial dose being administered anytime from the 16th to the 30th week of gestation up until the time of delivery. Follow-up of the seven infants from birth with a mean duration of 22 months (range 16-32 months) revealed mild drug-related toxicity: anemia in two infants and macrocytosis in all seven, both conditions resolved by the second month of life. All infants remained HIV-1 seronegative, according to the 1987 CDC classification, and all stayed clinically well. Other virological parameters including virus culture, in vitro antibody production, and polymerase chain reaction, repeatedly performed in the infants, remained negative. Although none of the mothers transmitted HIV-1 infection to the offspring, the size of this study and the relatively low transmission rate (13%) in Europe do not permit us to draw a definite conclusion about treatment efficacy in preventing maternal-fetal transmission. However, the drug caused only limited toxicity among the infants, and its administration to large numbers of mothers in treatment trials should be considered relatively safe for both mother and child.

Ten children affected by HBV chronic hepatitis, not responding to a previous treatment with interferon (IFN), have been treated with a reiterated IFN therapy. The response obtained is not encouraging and only one patient became negative for HBeAg and HBV-DNA.

In this preliminary study, children with chronic HBV hepatitis, as was also previously shown for adults, respond to interferon therapy in an HLA class I antigen dependent manner. If this can be confirmed on a large scale, HLA typing may serve as a useful indication of interferon-therapy responders.

The crude supernatant of an CD1+/CD8+ T-cell clone (GI-CO-T-9) established from a long standing culture of an acute T-lymphoblastic leukemia (T-ALL) was shown to inhibit the responsiveness of normal PBMC to PHA. This clone does not produce TNF-alpha, TNF-beta, alpha-IFN, tau-IFN, IL-1, IL-2 and has no NK-like activity. The crude supernatant has been subjected to a multi-step chromatographic fractioning. Preparative gel permeation HPLC allowed us to recover two peaks of biologic activity in the range of 100-120 kDa and 75-85 kDa (referred to as "high molecular mass inhibitor factor", HMMIF, and "low molecular mass inhibitor factor", LMMIF, respectively). Both fractions were then subjected to anion exchange HPLC: HMMIF was recovered in fractions eluting at 0.04 M NaCl while LMMIF eluted at higher ionic strength (0.48 M NaCl). The fractions with biologic activity recovered from anion exchange HPLC have been subjected to hydrophobic interaction HPLC (HIC) for final purification. The highly purified material was characterized by polyacrylamide gel electrophoresis with and without sodium dodecyl sulfate (SDS) revealing single bands of 115 kDa and 80 kDa. The isoelectric points (pI), determined by flat-bed isoelectricfocusing, were 7.4 for HMMIF and 3.5-3.6 for LMMIF. Studies on temperature lability indicated that both proteins are stable for 3-4 hours at room temperature (RT), 24-36 hours at +4 degrees C and 7-10 days at -80 degrees C.

The supernatant of CD8+ cells, isolated from a permanent lymphoblastoid cell clone established from a long term culture of a T cell acute lymphoblastic leukemia, contained two distinct molecules with suppressive activity on PHA1-induced PBMC proliferation. This clone does not produce TNF-alpha, TNF-beta, alpha-IFN, gamma-IFN, IL-1, IL-2 and has not natural killer activity. In the attempt to purify and biochemically characterize the lymphoblastic cell line-derived T-cell SFs, a multi-step chromatographic separation has been used. Two different peaks of biologic activity have been separated by HPLC gel permeation in the range of 100-120 Kd and 75-85 Kd referred to as high molecular weight suppressor factor HMWSF, and low molecular weight suppressor factor LMWSF, respectively. These fractions were then concentrated, dialyzed and further purified by anion exchange HPLC. This chromatographic step allowed us to considerably purify the two SFs. The biologically active fractions derived from the previous chromatographic step were eventually subjected to hydrophobic interaction HPLC (HIC) for final purification. The highly purified material was characterized by polyacrylamide gel electrophoresis in sodium-dodecylsulfate (SDS-PAGE): a single band corresponding to 115 Kd was observed for HMWSF, while LMWSF yielded a single band at 80 Kd. The isoelectric points (pI) of the different SFs was determined by flat-bed isoelectric-focusing: the HMWSF yielded a single band at pI 7.4, while a much lower pI was observed for LMWSF, 3.5-3.6. Studies on temperature lability indicated that both proteins are stable for 3-4 hours at room temperature (RT), 24-36 hours at +4 degrees C and 7-10 days at -80 degrees C.

Cytosine-arabinoside (ARA-C) effects on a new human neuroblastoma cell line (GI-ME-N), recently established in our laboratory, have been extensively tested. Low doses of ARA-C allowing virtually 100% cell viability induce morphological differentiation and growth inhibition; differentiated cells appear larger and flattened with elongated dendritic processes; such cells appeared within 48 hours after a dose of ARA-C as low as 0.1 microgram/ml. The new morphological aspect reached the maximum expression after 5-6 days of culture being independent of the addition of extra drug to the culture. A decrease in (3H)-thymidine incorporation was also observed within 48 hours being the cell growth completely inhibited at the 6th day. Membrane immunofluorescence with specific monoclonal antibodies showed several dramatic changes in NB-specific antigen expression after 5 days of treatment with ARA-C. These findings suggest that low ARA-C doses promote the differentiation of GI-ME-N neuroblastoma cells resulting in an altered expression of the malignant phenotype.

The aim of the present work is to assess whether reactivation of viral replication occurred in children affected by chronic hepatitis undergoing long term immunosuppressive therapy. 123 serum samples belonging to 25 children were retrospectively evaluated for HBV-DNA and HBeAg. Sera were collected prior to and during the protocol treatment (steroids alone or with azathioprine). Presence of HBV-DNA was evaluated by means of molecular hybridization technique, using a radiolabelled probe of cloned HBV-DNA. Sera (100 1) were denatured and transferred into nylon membrane (spot) then prehybridized and hybridized. After washing in stringent conditions, the filter was exposed in autoradiographic cassette with Kodak film XOmat5. Positivity was semiquantitatively evaluated by blackening of the spot. Increase or appearance of HBV-DNA was observed in the sera from 24/25 pts. HBeAg became positive in 4/5 pts previously negative. Obtained data shows reactivation of viral replication during immunosuppressive therapy. Data are especially significant in those cases in which such activity has apparently ceased or not been detected; HBV may be considered as a latent virus.

A cell clone (GI-CO-T-9) derived from a long term T-cell culture (PF-382), established from a patient affected by acute T-lymphoblastic leukemia (T-ALL), was selected for the presence in the culture medium of factors suppressing T-cell proliferation. The crude supernatant has been subjected to a multi-step chromatographic fractioning, including: preparative gel permeation, anion exchange, and hydrophobic interaction High Performance Liquid Chromatography (HPLC). The highly purified material was characterized by polyacrylamide gel electrophoresis in sodium dodecyl sulfate (SDS-PAGE), revealing single bands of 115 Kd and 80 Kd. The isoelectric points (pI), determined by flat-bed isoelectric-focusing, were 7.4 for High Molecular Weight Suppressor Factor (HMWSF) and 3.5-3.6 for Low Molecular Weight Suppressor Factor (LMWSF).

The efficacy of human lymphoblastoid interferon (Wellferon) therapy was measured in 20 children with chronic hepatitis B with or without pretreatment with prednisolone. Patients were randomised to receive 0.6 mg/kg/day prednisolone for 3 weeks, then at 0.3 mg/kg for a fourth week or placebo. All patients then received interferon 5 MU/m2 i.m. for 12 weeks; daily for 5 days then three times a week for the remaining 11 weeks. Preliminary results show that 25% of children had a permanent loss of viral markers of replication. However, response to interferon varied widely between individuals and a prolonged follow-up is required in order to determine the influence of prednisolone pretreatment on the efficacy of interferon therapy.