LDL-cholesterol lowering effect of a new dietary supplement: an open label, controlled, randomized, cross-over clinical trial in patients with mild-to-moderate hypercholesterolemia.

BACKGROUND: Hypercholesterolemia is a major risk factor for cardiovascular disorders and requires specific intervention through an adequate lifestyle (diet and physical exercise) and, if necessary, an appropriate drug treatment. Lipid-lowering drugs, although generally efficacious, may sometimes cause adverse events. A growing attention has been devoted to the correction of dyslipidemias through the use of dietary supplements. The aim of this study was to assess the lipid-lowering activity and safety of a dietary supplement containing monacolin K, L-arginine, coenzyme Q10 and ascorbic acid, named Argicolina (A), compared to a commercially available product containing monacolin K and coenzyme Q10, Normolip 5 (N). METHODS: This was a single center, controlled, randomized, open-label, cross-over clinical study enrolling 20 Caucasian outpatients aged 18-75 years with serum LDL-C between 130 and 180 mg/dL. Patients assumed two different dietary supplements (A and N) both containing monacolin K 10 mg for 8 weeks each, separated by a 4-week wash-out period. Evaluated parameters were: Total cholesterol (Tot-C), low density lipoprotein cholesterol (LDL-C), high density lipoprotein cholesterol (HDL-C), triglycerides (TG), fasting blood glucose, aspartate aminotransferase, alanine aminotransferase, creatinekinase, gamma-glutamyl-transpeptidase, brachial arterial pressure and heart rate, measured at the start and at the end of each treatment period. Safety was monitored through the study. RESULTS: LDL-C decreased by 23.3% during treatment with N (p < 0.0001) and by 25.6% during treatment with A (p < 0.0001); the LDL-C mean reduction was 36.4 (95% CI: 45,6-27,1) mg/dL during N treatment and 40.1 (95% CI: 49.2-30,9) mg/dL during A treatment. Tot-C decreased significantly (p < 0.0001) within each treatment period. HDL-C increase was negligible during A whereas it was significant during N. TG diminished markedly during A and not significantly during N. The difference between treatments was not statistically significant for all variables. No serious or severe adverse events occurred during the study. CONCLUSIONS: Our results confirm the clinically meaningful LDL-C lowering properties of monacolin K. At variance with a supplement already in the market (N), the novel association (A) of monacolin K with L-arginine, coenzime Q10 and ascorbic acid also produces a significant reduction of triglycerides without significant effects on HDL. TRIAL REGISTRATION: ClinicalTrials.gov ID: NCT03425630 .

The effect of particle size and thermal pre-treatment on the methane yield of four agricultural by-products.

One way to optimize methane production in anaerobic digestion plants is to substitute ligno-cellulosic by-products for crops traditionally used as energy sources. However, using these by-products requires introduction of a pre-treatment system to minimize energy input and maximize energy output for an improved net energy equation. In this study, four agricultural byproducts (wheat, barley, rice straw and maize stalks) underwent various mechanical and thermal treatments prior to anaerobic digestion including particle size reduction to 5.0, 2.0, 0.5, and 0.2 cm and heat application to 90 °C and 120 °C. Mechanical pre-treatment increased byproduct methane yields more than 80%; thermal pre-treatment improved yields more than 60% for wheat and barley straw. Pre-treating wheat straw improved methane yields most, regardless of whether the method was thermal or mechanical. An electric net energy balance was also completed to analyze the feasibility of the pre-treatments according to input and output of energy.

Mutations in the motor and stalk domains of KIF5A in spastic paraplegia type 10 and in axonal Charcot-Marie-Tooth type 2.

Spastic paraplegia type 10 (SPG10) is an autosomal dominant form of hereditary spastic paraplegia (HSP) due to mutations in KIF5A, a gene encoding the neuronal kinesin heavy chain implicated in anterograde axonal transport. KIF5A mutations were found in both pure and complicated forms of the disease; a single KIF5A mutation was also detected in a CMT2 patient belonging to an SPG10 mutant family. To confirm the involvement of the KIF5A gene in both CMT2 and SPG10 phenotypes and to define the frequency of KIF5A mutations in an Italian HSP patient population, we performed a genetic screening of this gene in a series of 139 HSP and 36 CMT2 affected subjects. We identified five missense changes, four in five HSP patients and one in a CMT2 subject. All mutations, including the one segregating in the CMT2 patient, are localized in the kinesin motor domain except for one, falling within the stalk domain and predicted to generate protein structure destabilization. The results obtained indicate a KIF5A mutation frequency of 8.8% in the Italian HSP population and identify a region of the kinesin protein, the stalk domain, as a novel target for mutation. In addition, the mutation found in the CMT2 patient strengthens the hypothesis that CMT2 and SPG10 are the extreme phenotypes resulting from mutations in the same gene.

The GST domain of GDAP1 is a frequent target of mutations in the dominant form of axonal Charcot Marie Tooth type 2K.

BACKGROUND: Mutations in GDAP1 associate with demyelinating (CMT4A) and axonal (CMT2K) forms of CMT. While CMT4A shows recessive inheritance, CMT2K can present with either recessive (AR-CMT2K) or dominant segregation pattern (AD-CMT2K), the latter being characterised by milder phenotypes and later onset. The majority of the GDAP1 mutations are associated with CMT4A and AR-CMT2K, with only four heterozygous mutations identified in AD-CMT2K. METHODS: We screened GDAP1 gene in a series of 43 index patients, 39 with CMT2 and 4 with intermediate CMT, with sporadic and familial occurrence of the disease. RESULTS: Three novel mutations were identified in three families with dominant segregation of the disease: two missense changes, p.Arg226Ser and p.Ser34Cys, affecting the GST domain of the GDAP1 protein and a novel deletion (c.23delAG) leading to early truncation of the protein upstream the GST domain. Wide variability in clinical presentation is shared by all three families mostly in terms of age at onset and disease severity. A rare variant p.Gly269Arg, located within the GST domain, apparently acts as phenotype modulator in the family carrying the deletion. CONCLUSION: The results obtained reveal a GDAP1 mutation frequency of 27% in the dominant families analysed, a figure still unreported for this gene, thus suggesting that GDAP1 involvement in dominant CMT2 might be higher than expected.

Mutated mitofusin 2 presents with intrafamilial variability and brain mitochondrial dysfunction.

BACKGROUND: The axonal forms of Charcot-Marie-Tooth (CMT2) disease are a clinically and genetically heterogeneous group of disorders. Mitofusin 2 gene (MFN2) mutations are the most common cause of CMT2. Complex phenotypes have been described in association with MFN2 gene mutations, including CMT2 with pyramidal features (hereditary motor and sensory neuropathy [HSMN V]) and CMT2 with optic atrophy (HMSN VI). OBJECTIVE: To report on the clinical, neurophysiologic, and neuropathologic features of an Italian family with a novel MFN2 gene mutation and investigate brain functional parameters using magnetic resonance spectroscopy (MRS). METHODS: Three family members, a father and his two sons, were affected by peripheral neuropathy, cognitive impairment, and poor nocturnal vision (also optic neuropathy in one case). A member of this family also showed spastic paraparesis. The MFN2 gene sequence was analyzed. A sural nerve biopsy as well as brain (1)H-MRS and (31)P-MRS were evaluated in two patients. RESULTS: Affected family members carried a novel MFN2 missense mutation, namely R104W, located within the critical GTPase domain of the protein which affects a highly conserved amino acid position. Sural nerve biopsies showed a normal mitochondrial network, particularly at the nodes of Ranvier, upon electron microscopy examination. A significant defect of high energy phosphates (HEPs) in the visual cortex was observed at rest by (31)P-MRS in the adult proband, while his son showed a defective recovery of HEPs after stimulation of the visual cortex. CONCLUSION: Cognitive impairment may be another feature of the MFN2-related phenotype. The widespread peripheral and CNS involvement, as well as the neurosensorial defects, underline the similarities among MFN2-related and primary mitochondrial disorders.

Suspected acute allergic reactions: analysis of admissions to the Emergency Department of the AOU Maggiore della Carità Hospital in Novara from 2003 to 2007.

OBJECTIVE OF THE STUDY: The aim of our work is to ascertain the frequency and the impact of acute allergic reactions on the routine of a highly-specialized Emergency Department collecting information on the admission, the typology of symptoms and the degree of severity calculating the incidence and the outcomes of the events. MATERIALS AND METHODS: The study started the 1 July 2006 and the records of the Emergency Department of the Maggiore della Carità Hospital in Novara were consulted retrospectively in the period between the 1 January 2003 and the 31 December 2006, and prospectively up to the 31 December 2007, using keywords that could identify admission for suspected allergic reactions. Information relating to internal medicine and/or pediatric cases were examined, excluding all surgical and/or trauma cases. The number ofadmissions per year was considered broken down by clinical signs, triage assessment upon admission and discharge outcome. RESULTS: Admissions to the Emergency Department during the period under consideration were 165,120 with 6107 suspected cases of allergic reactions. The symptoms most frequently reported both in adults (A) and children (C < or =18 years old), were: hives 37%, asthma 20.65 (A)% and 27.4% (C); drug allergy 7.5% (A) and 6.1% (C). Reactions to Hymenoptera venom were less frequent, 4.7% (A) and 1.27% (C); the frequency of angioedema, conjunctivitis and rhinitis was between 1 and 4%. The incidence of food allergies (1.4%) and anaphylaxis (0.8%) was comparable for all ages. The triage assessment showed a significant percentage of "yellow" and "red" codes, with 362 cases (5.9%) and 71 cases (1.16%) respectively. A total of 151 patients was hospitalized, no one classified as "white" code. Death occurred in 7 cases: 4 "yellow" codes and 3 "red" codes, respectively. A more detailed specialistic evaluation was recommended in only 10% of the patients. CONCLUSIONS: Admissions to the Emergency Department for suspected allergic reaction are proportional to the number of overall admissions for internal medicine cases and do not appear to be related to the general increase of allergies in the population. This led us to focus our attention on how allergic diseases impact the work of an Emergency Department and how to describe the discharge diagnosis better. A significant number of descriptive diagnoses also turned out to be inaccurate and did not allow the syndrome to be identified properly. The analysis of this information aims to be a stimulus to improve the emergency clinical approach used for allergic diseases and to plan the adequate management ofallergic patients after they have been treated in hospital.

Amino acid changes in the amino terminus of the Na,K-adenosine triphosphatase alpha-2 subunit associated to familial and sporadic hemiplegic migraine.

Familial hemiplegic migraine (FHM) is a rare subtype of migraine with aura inherited with an autosomal dominant pattern. Here, we report the genetic analysis of four families and one sporadic case with hemiplegic migraine (HM) in whom we searched for mutations in the three genes associated with the disease CACNA1A, ATP1A2 and SCN1A. Two novel amino acid changes p.Arg65Trp and p.Tyr9Asn, in the Na,K-adenosine triphosphatase (ATPase) alpha-2 subunit encoded by the ATP1A2 gene, were found in one FHM family and in the sporadic case, respectively. These mutations are peculiar for their location in the extreme N-terminus, an uncommon mutation target in this protein. Low frequency of migraine attacks in all our mutant patients with low complexity of the associated aura symptoms in the sporadic case is also observed. Besides the two novel mutations, the data here reported confirm the involvement of ATP1A2 gene in the sporadic form of HM, while the negative results on the other families tested for all genes known in HM strengthen the hypothesis of the existence of at least another locus involved in FHM.

Improved recycling of livestock slurries on maize by means of a modular tanker and spreader.

Slurry spreading represents a critical component of the waste management strategy since it influences the nutrients' availability for the crops. The Italian Regulation in force sets severe limits to the slurry application to land. These limits--and the necessity to optimise the slurry fertilising value--require to operate with the correct application rate and with a sufficient spreading uniformity. An innovative slurry spreader able to optimise the reuse of this by-product as an agronomic resource whilst respecting the national regulation has been tested under practical conditions. The agronomic trials showed that the controlled recycling--performed with the innovative slurry spreader--gave no statistically different grain yields from those obtained with the chemical fertilisation. The ammonia emissions recorded after the slurry injection were the 16% of those recorded after the band application.

[Nephrotic syndrome refractory to conventional therapy]. / Sindrome nefrosica refrattaria alla terapia convenzionale.

BACKGROUND: Prednisone is the choice medicine in Nephrotic Syndrome (NS) treatment, possibly associated with immunosuppressor medicines (cyclophosphamide or chlorambucil), either in case of NS resistance at cortisone therapy or with frequent relapses. Cyclosporin A (CyA) use has been recently proposed, due to its inhibitory effect on the IL2 and lymphokine release, with a permeabilizing effect on the glomerular membrane. The purpose of this study is to evaluate the CyA antiproteinuric effectiveness with NS conventional therapy refractory patients. METHODS: Six patients (3 females and 3 males) have been treated with CyA (4 +/- 0.5 mg/Kg/die) associated with low corticosteroid dosages. RESULTS: During the treatment, proteinuria reduced in 5 patients, at less than 1/3 of pre-treatment values, for 4 patients this happened starting from the 2nd month of therapy, while after the 12th for the fifth patient. The sixth patient has now a 2/3 reduction compared to the initial one and he is at the 3rd month of therapy. During the CyA treatment, further to the proteinuria reduction, a total proteinemia values increase and a cholesterolemia and tryglyceridemia reduction has been observed, while creatinine and PA have not changed. CONCLUSIONS: Four out of the six treated patients have been respectively under therapy for 2,3,12,30 months. Two stopped CyA therapy: one after 18 months due to clinical stability, still present after 2 years from interruption; one after 9 months with a stable clinical picture for just three months, since she was longing for a pregnancy, achieving a quick proteinuria relapse.

Usefulness of transdermal clonidine in hypertensive patients undergoing minor surgical operations.

Transdermal clonidine (TTSC) treatment was evaluated in 29 patients with mild to moderate hypertension scheduled for minor surgery. Two weeks before the scheduled operation, patients underwent 24-h ambulatory blood-pressure monitoring (ABPM) to evaluate the efficacy of previous oral antihypertensive treatment, which was then substituted with TTSC, 0.1 mg/day. After 1 week, the efficacy of TTSC was clinically assessed, and the dose increased to 0.2 mg/day if needed. ABPM was repeated 2 days before the scheduled operation and 2 days after surgery. The 24-h blood pressure (BP) and heart rate (HR) profiles were smoothed by Fourier analysis. Three patients withdrew for adverse events and one for inefficacy after dose adjustment, TTSC being effective in the remaining 25 patients. Two patients who completed treatment lacked postsurgical ABPM recording. In the 23 patients with all ABPM recordings, average 24-h BP and HR obtained preoperatively during TTSC treatment were slightly reduced compared with values recorded during previous oral therapy. BP changes after surgery were negligible, whereas HR showed a moderate increase. Minor adverse events occurred in four (14%) of 29 patients. Our results demonstrate that TTSC provides adequate BP control in patients with mild to moderate hypertension undergoing minor surgery.

Oral environment temperature changes induced by cold/hot liquid intake.

The use of NiTi shape memory alloys, introduced into orthodontics because of their ability to develop light continuous forces that prove more effective than heavy intermittent forces in the teeth movement, requires the mastering of the functional properties of NiTi wires. More specifically, the recovery force acting on the teeth is a sensitive function of temperature: knowledge of oral temperature modifications is therefore required to understand the stress state modification felt during orthodontic therapy. The temperature modifications induced by cold or hot drink intake in the oral cavity were investigated by using arch wires, fixed to removable Hawley retainers, similar to those currently used in orthodontic practice, by means of six temperature sensors placed in correspondence with specific teeth. Similarly, the temperature changes were detected on a metallic frame, fixed onto the palatal zone to a Hawley retainer, where a palatal expander was placed to correct unilateral or bilateral crossbites in deciduous or in early mixed dentition. The maximum temperature change was observed in the interincisor area: The temperature modification on other teeth depends on the modality of drink intake, with the highest temperature variations being detected in the palatal zone. Hence modifications in the stress state during orthodontic therapy with NiTi wires are to be expected.

Description of mandibular finite helical axis pathways in asymptomatic subjects.

Despite wide use of systems to record jaw motion with six degrees of freedom, most studies have analyzed only the movement of a single mandibular point. The finite helical axis (FHA) is a mathematical model which can be used to describe comprehensively the movements of a rigid body. The aim of this investigation was to describe the FHA of the mandible during habitual jaw movements. Thirty subjects (13 females, 17 males; mean age, 26 years; range, 18 to 34 years) without myoarthropathies of the masticatory system participated in the study. Opening and closing movements, performed at 1-Hz frequency, were recorded with the optoelectronic system Jaws-3D. Three opening and closing movements were recorded from the right side and three from the left side of the jaw. The movement data were low-pass-filtered for noise reduction prior to the computation of the finite helical axis by means of a software program developed in our laboratory. The following parameters were calculated: the rotation of the FHA, its spatial orientation, and the translation along it, as well as its position and distance relative to an intracondylar point. In addition, methodological errors of the model were calculated. During opening and closing, the group mean FHA rotation was 24.3 degrees +/- 4.2 degrees. The group mean of the maximum total translation along the FHA was 0.9 +/- 0.7 mm. The group mean distance between the FHA and the intracondylar point was 48.9 +/- 9.9 mm. The FHA pathways were smooth and varied between individuals. Furthermore, the finite helical axes were never localized within the condyle, and often were located outside of the mandible. The analysis of the FHA pathways yields more information on whole mandibular movements than simply the movements of a single condylar point.

[Medical nephropathies: what has changed in 20 years]. / Nefropatie mediche: cosa è cambiato in 20 anni.

The authors analyse the series of patients with medical nephropathy undergoing renal biopsy between 1973 and 1993 in order to make a diagnostic and prognostic comparison between the first (ID) and second (IID) decade. Clinical indications for biopsy, which became more precise during the second decade, led to the diagnosis of fewer patients with normal histology; the introduction of ME and IF allowed non-significant histological conditions to be reduced during IID; echo-guided biopsy has led to a reduced number of post-biopsy complications in IID compared to ID. Epidemiological analysis reveals the reduction of focal glomerulosclerosis in IID in favour of glomerulonephritis with IgA deposits in correlation with the use of IF; the increase in mebranous glomerulonephritis secondary to increased antigenic stimuli; reduced acute post-infective glomerulonephritis and membrane-proliferative glomerulonephritis owing to an improved prophylaxis of sources of infection. Among the patients undergoing renal biopsy and commencing dialysis an increase was observed in IID in the number of cases of membranous glomerulonephritis or caused by IgA deposits. There was an increased interval between biopsy and the start of dialysis in IID compared to ID, in spite of fewer patients receiving immunosuppressive therapy. This was probably due to the increased number of pathologies with a slower evolution, thus justifying the postponement of the start of dialysis.

Innovative materials: the NiTi alloys in orthodontics.

Since ten years the NiTi alloys have gained an ever increasing place in orthodontic practice: that is due to their peculiar mechanical properties ascribed to a martensitic thermoelastic transformation which can be thermally or, in a proper temperature range, stress-induced. In the last case, when martensite is stress-induced at body temperature, the stress-strain behaviour is pseudoelastic with large deformations gained or recovered at constant stress, respectively in direct/reverse transformation: this behaviour exploited in orthodontics allowed to overcome the drawbacks intrinsic to the use of conventional alloys as stainless steel or Co-Mo alloys, where small displacements can be achieved at decreasing loads. From the phase state diagram of NiTi alloys it appears that at body temperature they are stable, but out of equilibrium: thermal treatments at intermediate temperatures can therefore modify the equilibrium state and as a consequence the transformation temperatures respect to body temperature. That allows to modify the recovery stress level according to the requirements of practice and thus disclosing new roads: the capability to foresee NiTi archwires pre-programmed in different sections, with a personalized scheme. Attention has not currently been paid to the modifications in the recovery stress induced by a temperature change inside the oral cavity. Recent results have shown that the thermal changes in the oral cavity induced by cold/hot liquid intake can considerably modify the stress level to which the dentition is exposed: though confined to the time extent connected with drinking, similar effects can be expected also for meals intake and should be taken into account for a correct procedure.

Reliability of immunoassays for anti-HCV antibodies (ELISA and RIBA II) in patients with essential mixed cryoglobulinemia (EMC).

The recent reports of a very high frequency of signs of hepatitis C virus (HCV) infection among patients with essential mixed cryoglobulins (EMC) suggest new hypotheses for the pathogenesis of this disease. However, most of these studies have been seriously criticized. The serologic test designed for detection of anti-HCV antibodies (ELISA, RIBA I and II) may yield a significant rate of false-positive results when performed on cryoglobulinemic sera, and the detection of the HCV genome by PCR is still heavily conditioned by practical problems. Indirect, but possibly more reliable, evidence of HCV infection in cryoglobulinemic patients might come from the demonstration of anti-HCV antibodies by a conventional technique (ELISA or RIBA) in the purified polyclonal non-rheumatoid immunoglobulinemic fraction excreted in the urine by glomerular filtration. Fifty-two patients whose serum had tested positive for HCV antibodies (by ELISA and RIBA) on multiple occasions were enrolled in this study. They were diagnosed as having either EMC or HCV chronic hepatitis without cryoglobulinemia at least one year ago. The urine samples of these patients were tested for anti-HCV antibodies by ELISA and RIBA. In patients with chronic C hepatitis the antibodies most frequently found in the serum were anti-C33c and anti-C22-3. The results of the RIBA were substantially confirmed by ELISA, with a positive test in the urine of 30 of 32 seropositive patients. Similar results were obtained in patients with EMC II. We conclude that the specificity of the RIBA and ELISA tests for HCV antibodies in patients with EMC appears to be as high as in HCV+ patients without serum cryoglobulins. EMC patients have a high incidence of HCV infection and active chronic liver disease.

[Microalbuminuria: theoretical bases and new applications]. / La microalbuminuria: basi teoriche ed aspetti applicativi.

The constant presence of albumin, as detected by common biochemical methods, in multiple urine samples of a patient, was first considered by Bright, in 1836, as a cardinal sign of renal disease ("clinical proteinuria"). Since then this view was widely adopted for studying the clinical evolution of the patients with diabetes mellitus, whose high risk to develop proteinuria and subsequently a progressive decline of renal function was well known. Thus the finding of "clinical proteinuria" by traditional, merely biochemical techniques, has been considered for more than one century as the opening event in the onset of diabetic nephropathy, and a distinctive sign of glomerulopathy in general. More recently, this view has been deeply criticized, mainly because it lies on the implicit assumption that the sensitivity limits of the biochemical tests for the detection of urinary protein concentrations (about 300 mg/dl), coincide with the ones that can distinguish non nephropathic from nephropathic patients (either diabetic or not). Indeed new techniques, that detect urinary proteins down to 1 microgram/ml, have shown that the upper limit of protein excretion in healthy people is well below the minimum concentration detectable by all the traditional tests. Therefore a new clinical entity, named "microproteinuria" has been defined, meaning the urinary excretion rate ranging between the "physiological" and the "clinical" proteinuria; its pathophysiologic, diagnostic and prognostic significance has been extensively evaluated in the last 20 years. Microproteinuria has been shown to represent a crucial event in the natural history of the diabetic nephropathy; in diabetic patients it is strictly related to the risk of future (months to years) development of overt nephropathy and chronic renal failure, and it may predict the risk of macroangiopathic complications. More recently new settings have been proposed for the study of microproteinuria, as an early and sensitive marker of cardiovascular diseases in hypertensive non diabetic patients and even in non hypertensive non diabetic elderly people. The role of microproteinuria in the diagnosis and follow-up of many non-diabetic glomerulopathies is a very interesting though still unexplored field.

Effect of plasma exchange in a patient with prolonged deep coma from thrombotic thrombocytopenic purpura.

The clinical, laboratory and instrumental data of a patient with thrombotic thrombocytopenic purpura (TTP) are reported in detail, with reference to an articulated therapeutic regimen including plasma exchange, high-dose methylprednisolone and dipiridamole. Particular emphasis is placed on the dramatic improvement obtained with this treatment despite a very delayed diagnosis and prolonged, severe neurological involvement. A short discussion and review of the literature concerning the role of plasma-exchange in the treatment of TTP is reported.

Parenchymal renal involvement in three cases of non-Hodgkin lymphomas: clinical and pathological features.

Parenchymal neoplastic invasion of the kidneys is a common postmortem finding in patients who have died from advanced non Hodgkin's lymphomas (NHL). However, it rarely causes major clinical consequences, such as impairment of glomerular and tubular function, acute or rapidly progressive renal failure. Renal involvement is even less frequent as a first manifestation of NHL, the so-called "primary" renal lymphoma. A review of the main clinical, diagnostic and pathological aspects of three cases observed in our division is presented here.

Hepatitis C virus infection in type II essential mixed cryoglobulinemias.

The possible relationship between essential mixed cryoglobulinemias (EMCs) and hepatitis C virus (HCV) has been investigated in eight patients with type II EMCs and biochemical signs of liver damage, whose serum tested positive in the ELISA for anti-HCV. Sera were tested using the 2nd generation RIBA assay, while serum HCV-RNA was measured semiquantitatively by a RT-PCR in whole serum, cryoprecipitates and supernatants. In all patients a percutaneous liver biopsy and a bone marrow biopsy were performed. At liver biopsy, chronic active hepatitis and/or cirrhosis were present in 6 patients; in the remaining two, a lymphoplasmacytoid infiltration of elements positive for kappa light chains was found. In all patients a bone marrow biopsy showed a paratrabecular infiltration of monoclonal lymphoplasmacytoid elements similar to those found in the liver of the two patients described above. Antibodies against structural and non-structural HCV proteins were detectable in the serum of all patients. HCV-RNA was amplified from the whole sera, cryoprecipitates and supernatants: significantly higher concentrations were found in cryoprecipitates than in supernatants. Our results confirm the high prevalence of HCV infection and ongoing viral replication in patients with type II EMC and suggest the possible implication of HCV in EMC pathogenesis.