[SAMHD1 acts at stalled replication forks to prevent interferon induction]. / SAMHD1 agit sur les fourches de réplication bloquées pour empêcher l'induction d'interféron.

DNA replication is an extremely complex process, involving thousands of replication forks progressing along chromosomes. These forks are frequently slowed down or stopped by various obstacles, such as secondary DNA structures, chromatin-acting proteins or a lack of nucleotides. This slowing down, known as replicative stress, plays a central role in tumour development. Complex processes, which are not yet fully understood, are set up to respond to this stress. Certain nucleases, such as MRE11 and DNA2, degrade the neo-replicated DNA at the level of blocked forks, allowing the replication to restart. The interferon pathway is a defense mechanism against pathogens that detects the presence of foreign nucleic acids in the cytoplasm and activates the innate immune response. DNA fragments resulting from genomic DNA metabolism (repair, retrotransposition) can diffuse into the cytoplasm and activate this pathway. A pathological manifestation of this process is the Aicardi-Goutières syndrome, a rare disease characterized by chronic inflammation leading to neurodegenerative and developmental problems. In this encephalopathy, it has been suggested that DNA replication may generate cytosolic DNA fragments, but the mechanisms involved have not been characterized. SAMHD1 is frequently mutated in the Aicardi-Goutières syndrome as well as in some cancers, but its role in the etiology of these diseases was largely unknown. We show that cytosolic DNA accumulates in SAMHD1-deficient cells, particularly in the presence of replicative stress, activating the interferon response. SAMHD1 is important for DNA replication under normal conditions and for the processing of stopped forks, independent of its dNTPase activity. In addition, SAMHD1 stimulates the exonuclease activity of MRE11 in vitro. When SAMHD1 is absent, degradation of neosynthesized DNA is inhibited, which prevents activation of the replication checkpoint and leads to failure to restart the replication forks. Resection of the replication forks is performed by an alternative mechanism which releases DNA fragments into the cytosol, activating the interferon response. The results obtained show, for the first time, a direct link between the response to replication stress and the production of interferons. These results have important implications for our understanding of the Aicardi-Goutières syndrome and cancers related to SAMHD1. For example, we have shown that MRE11 and RECQ1 are responsible for the production of DNA fragments that trigger the inflammatory response in cells deficient for SAMHD1. We can therefore imagine that blocking the activity of these enzymes could decrease the production of DNA fragments and, ultimately, the activation of innate immunity in these cells. In addition, the interferon pathway plays an essential role in the therapeutic efficacy of irradiation and certain chemotherapeutic agents such as oxaliplatin. Modulating this response could therefore be of much wider interest in anti-tumour therapy.

La réplication de l'ADN est un processus extrêmement complexe, impliquant des milliers de fourches de réplication progressant le long des chromosomes. Ces fourches sont fréquemment ralenties ou arrêtées par différents obstacles, tels que des structures secondaires de l'ADN, des protéines agissant sur la chromatine ou encore un manque de nucléotides. Ce ralentissement, qualifié de stress réplicatif, joue un rôle central dans le développement tumoral. Des processus complexes, qui ne sont pas encore totalement connus, sont mis en place pour répondre à ce stress. Certaines nucléases, comme MRE11 et DNA2, dégradent l'ADN néorépliqué au niveau des fourches bloquées, ce qui permet le redémarrage des réplisomes. La voie interféron est un mécanisme de défense contre les agents pathogènes qui détecte la présence d'acides nucléiques étrangers dans le cytoplasme et active la réponse immunitaire innée. Des fragments d'ADN issus du métabolisme de l'ADN génomique (réparation, rétrotransposition) peuvent diffuser dans le cytoplasme et activer cette voie. Une manifestation pathologique de ce processus est le syndrome d'Aicardi-Goutières, une maladie rare caractérisée par une inflammation chronique générant des problèmes neurodégénératifs et développementaux. Dans le cadre de cette encéphalopathie, il a été suggéré que la réplication de l'ADN pouvait générer des fragments d'ADN cytosoliques, mais les mécanismes impliqués n'avaient pas été caractérisés. SAMHD1 est fréquemment muté dans le syndrome d'Aicardi-Goutières ainsi que dans certains cancers, mais son rôle dans l'étiologie de ces maladies était jusqu'à présent largement inconnu. Nous montrons que de l'ADN cytosolique s'accumule dans les cellules déficientes pour SAMHD1, particulièrement en présence de stress réplicatif, activant la réponse interféron. Par ailleurs, SAMHD1 est important pour la réplication de l'ADN en conditions normales et pour le processing des fourches arrêtées, indépendamment de son activité dNTPase. De plus, SAMHD1 stimule l'activité exonucléase de MRE11 in vitro. Lorsque SAMHD1 est absent, la dégradation de l'ADN néosynthétisé est inhibée, ce qui empêche l'activation du checkpoint de réplication et entraine un défaut de redémarrage des fourches de réplication. De plus, la résection des fourches de réplication est réalisée par un mécanisme alternatif qui libère des fragments d'ADN dans le cytosol, activant la réponse interféron. Les résultats obtenus montrent, pour la première fois, un lien direct entre la réponse au stress réplicatif et la production d'interférons. Ces résultats ont des conséquences importantes dans notre compréhension du syndrome d'Aicardi Goutières et des cancers liés à SAMHD1. Par exemple, nous avons démontré que MRE11 et RECQ1 sont responsables de la production des fragments d'ADN qui déclenchent la réponse inflammatoire dans les cellules déficientes pour SAMHD1. Nous pouvons donc imaginer que bloquer l'activité de ces enzymes pourrait diminuer la production des fragments d'ADN et, in fine, l'activation de l'immunité innée dans ces cellules. Par ailleurs, la voie interférons joue un rôle essentiel dans l'efficacité thérapeutique de l'irradiation et de certains agents chimiothérapiques comme l'oxaliplatine. Moduler cette réponse pourrait donc avoir un intérêt beaucoup plus large en thérapie anti-tumorale.

Polystyrene nanoparticles may affect cell mitosis and compromise early embryo development in mammals.

A great interest surrounds the development of nanoparticles (NPs) for biomedical applications such as drug delivery and cancer therapy. However, the interplay between nanoscale materials and biological systems and the associated hazards have not been completely clarified yet. In this study, bovine oviductal epithelial cells (BOECs) and embryos were used as in vitro models to investigate whether cell mitosis and early mammalian embryo development could be affected by the exposure to polystyrene (PS) nanoparticles. Analysis of the karyotype performed on BOECs exposed to PS-NPs did not show chromosomal anomalies compared to the control, although more tetraploid metaphase plates were observed in the former. In vitro fertilization experiments designed to understand whether exposure to PS-NPs could affect pre-implantation development showed that incubation with PS-NPs decreased 8-cell embryo and blastocyst rate in dose-dependent fashion. The quality of the blastocysts in terms of mean cell percent blastomeres with fragmented DNA was the same in exposed blastocysts compared to controls. These results show that the exposure to PS-NPs may impair development. In turn, this may affect the rate of mitosis in embryos and yield a lower developmental competence to reach the blastocyst stage. This suggests that release in the environment and the subsequent accumulation of PS-NPs into living organisms should be carefully monitored to prevent cytotoxic effects that may compromise their reproduction rates.

In ataxia-teleangiectasia betamethasone response is inversely correlated to cerebellar atrophy and directly to antioxidative capacity.

BACKGROUND AND PURPOSE: Ataxia-telangiectasia (A-T) is a rare autosomal recessive disorder caused by alterations of the A-T mutated (ATM) gene. Although A-T is a non-curable disease, we, previously, documented a clear improvement of cerebellar functions during a short-term betamethasone trial. The aim of this study was to define the underlying biochemical mechanism. METHODS: In six A-T patients receiving a short-term steroid therapy, intracellular glutathione (GSH) levels were evaluated with a colorimetric assay. The lipid peroxidation level and reactive oxygen species (ROS) production were evaluated using commercial assays. All the parameters were compared with the improvement of cerebellar functions expressed as delta (Delta) of the Scale for the Assessment and Rating of Ataxia (SARA). RESULTS: We observed an inverse correlation between Delta SARA and the severity of cerebellar atrophy and between the latter and basal GSH values. Four of the five patients with the highest Delta SARA also had the highest GSH values. Moreover, even though basal ROS values were comparable in patients and controls, in the only patient studied at different time-points of therapy, a remarkable reduction in ROS levels was documented. CONCLUSION: We suggest that antioxidative mechanisms play a role in favouring the improvement of cerebellar functions observed in A-T patients receiving a short-term betamethasone trial.

Non-scheduled consultation in oncologic patients. How many of them are true emergencies? An observational prospective study.

Oncologic emergencies have been extensively described and clearly defined. In oncology daily practice, cancer patients seek non-scheduled medical care in situations they perceive as a medical emergency, but which may not be a true emergency. The aim of the study was to identify the main symptoms leading to a non-scheduled consultation (NSC) and their relationship to the type of cancer, and to evaluate whether the diagnosis at discharge of patients admitted as result of a NSC correlates with a true oncologic emergency. This was a prospective observational study. Between July 2002 and April 2003, 365 NSCs were recorded. The most frequent baseline diseases were breast cancer (70), lung cancer (67), gastrointestinal cancer (52), lymphoma (42) and ovarian cancer (22). The most common symptoms for consultation were: fever (84), pain (81), cutaneous manifestations (26), dyspnea (23), bleeding (16) and abdominal distention (16). Overall, 114 of 365 NSCs (31%) resulted in admission. The most frequent symptoms resulting in admission were fever (42), pain (16), dyspnea (11), vomiting (9), neurologic manifestations (7), abdominal distention (6) and anuria (6). At discharge, only 30 patients (26%) admitted after a NSC were diagnosed with a defined oncologic emergency: febrile neutropenia (13), intestinal occlusion (12), obstructive uropathy (4) and abdominal perforation (1). True emergencies were not the most frequent causes of NSC at our institution.

Reacciones agudas de hipersensibilidad secundarias a la infusión de quimioterapia ambulatoria: experiencia del Instituto Alexander Fleming / Secondary hypersensitivity reactions to antineoplastic agents: report of Instituto Alexander Fleming (Argentina)

Objetivo: Evaluar retrospectivamente las características de nuestra población, sus patologías prevalentes y las drogas recibidas; evaluar la naturaleza de las reacciones agudas de hipersensibilidad (RAH) producidas por la infusión de agentes antineoplásicos; evaluar las maniobras médicas instauradas y la necesidad de que estos tratamientos sean realizados en el ámbito hospitalario. Material y métodos: Se consideraron los pacientes (ptes.) que concurrieron a hospital de día para recibir tratamiento ambulatorio; se categorizaron las reacciones agudas en severas (requieren maniobras médicas activas para reestablecer la estabilidad clínica y se debe suspender la infusión) y moderadas (requieren maniobras médicas activas pero se puede reiniciar la infusión) sin considerarse las reacciones leves (por ej.: acatisia, eritema facial) ni las extravasaciones, se categorizaron las maniobras médicas en complejas (inotrópicos, internación) y no complejas (corticoides, antihistamínicos, alta institucional)...

Reacciones agudas de hipersensibilidad secundarias a la infusión de quimioterapia ambulatoria: experiencia del Instituto Alexander Fleming / Secondary hypersensitivity reactions to antineoplastic agents: report of Instituto Alexander Fleming (Argentina)

Objetivo: Evaluar retrospectivamente las características de nuestra población, sus patologías prevalentes y las drogas recibidas; evaluar la naturaleza de las reacciones agudas de hipersensibilidad (RAH) producidas por la infusión de agentes antineoplásicos; evaluar las maniobras médicas instauradas y la necesidad de que estos tratamientos sean realizados en el ámbito hospitalario. Material y métodos: Se consideraron los pacientes (ptes.) que concurrieron a hospital de día para recibir tratamiento ambulatorio; se categorizaron las reacciones agudas en severas (requieren maniobras médicas activas para reestablecer la estabilidad clínica y se debe suspender la infusión) y moderadas (requieren maniobras médicas activas pero se puede reiniciar la infusión) sin considerarse las reacciones leves (por ej.: acatisia, eritema facial) ni las extravasaciones, se categorizaron las maniobras médicas en complejas (inotrópicos, internación) y no complejas (corticoides, antihistamínicos, alta institucional)...(AU)

[Percutaneous lithotripsy: how to make it safer. Personal experience]. / Litolapassi percutanea: come renderla più sicura. Nostra esperienza.

BACKGROUND: Percutaneous surgery (PCN) is now a routinary method for the treatment of the majority of renal stones, since it has become safer than in the past, because, thanks to new endourologic instruments and to ultrasounds, it has been possible to reduce the mistakes of the renal puncture. Furthermore, the use of balloons catheters for the dilatation of the nephrostomic channel allows the reduction of operating time and hemorrhage risk. In this paper, the authors expose their experience in PCN operations carried out with the help of X-ray and ultrasonography, during the indirect laying of the stones, and the use of a balloon catheter for the creation of a working channel. METHODS: The Authors report their experience with the use of ultrasounds and concomitant X-rays for renal puncture, and of balloon catheter as track dilator, during 68 consecutive PCN carried out for renal lithiasis, 55 primary and 13 secondary to ESWL treatments; every stone has been cracked by a "Swiss-lithoclast" balistic lithotripter. For every patient, time of operation, complications and hospitalization-days were registered. RESULTS: Only three patients (4.4%) had haemorrhages and in one case of A-V fistula nephrectomy was necessary. The patients stayed in hospital approx. four days; the nephrostomic drainage was generally removed 3 days after the operation. CONCLUSIONS: The very low incidence of complications and the very short time of hospitalization suggest that ultrasounds and balloon-catheters may be useful to this surgery and may make it safer than in the past. Moreover, ultrasonography reduces the rate of X-rays exposition for operators and patients; the cost of the balloon is easily balanced by the reduction of operating times and hospitalization-days.

Mre11 protein complex prevents double-strand break accumulation during chromosomal DNA replication.

Mre11 complex promotes repair of DNA double-strand breaks (DSBs). Xenopus Mre11 (X-Mre11) has been cloned, and its role in DNA replication and DNA damage checkpoint studied in cell-free extracts. DSBs stimulate the phosphorylation and 3'-5' exonuclease activity of X-Mre11 complex. This induced phosphorylation is ATM independent. Phosphorylated X-Mre11 is found associated with replicating nuclei. X-Mre11 complex is required to yield normal DNA replication products. Genomic DNA replicated in extracts immunodepleted of X-Mre11 complex accumulates DSBs as demonstrated by TUNEL assay and reactivity to phosphorylated histone H2AX antibodies. In contrast, the ATM-dependent DNA damage checkpoint that blocks DNA replication initiation is X-Mre11 independent. These results strongly suggest that the function of X-Mre11 complex is to repair DSBs that arise during normal DNA replication, thus unraveling a critical link between recombination-dependent repair and DNA replication.

Role for cyclin-dependent kinase 2 in mitosis exit.

Mitosis requires cyclin-dependent kinase (cdk) 1-cyclin B activity [1]. Exit from mitosis depends on the inactivation of the complex by the degradation of cyclin B [2]. Cdk2 is also active during mitosis [3, 4]. In Xenopus egg extracts, cdk2 is primarily in complex with cyclin E, which is stable [5]. At the end of mitosis, downregulation of cdk2-cyclin E activity is accompanied by inhibitory phosphorylation of cdk2 [6]. Here, we show that cdk2-cyclin E activity maintains cdk1-cyclin B during mitosis. At mitosis exit, cdk2 is inactivated prior to cdk1. The loss of cdk2 activity follows and depends upon an increase in protein kinase A (PKA) activity. Prematurely inactivating cdk2 advances the time of cyclin B degradation and cdk1 inactivation. Blocking PKA, instead, stabilizes cdk2 activity and inhibits cyclin B degradation and cdk1 inactivation. The stabilization of cdk1-cyclin B is also induced by a mutant cdk2-cyclin E complex that is resistant to inhibitory phosphorylation. P21-Cip1, which inhibits both wild-type and mutant cdk2-cyclin E, reverses mitotic arrest under either condition. Our findings indicate that the proteolysis-independent downregulation of cdk2 activity at the end of mitosis depends on PKA and is required to activate the proteolysis cascade that leads to mitosis exit.

Responses to DNA damage in Xenopus: cell death or cell cycle arrest.

Xenopus embryos divide rapidly following fertilization. During this rapid period of cleavage, cell divisions are not sensitive to DNA replication or spindle assembly inhibition. Here, we have investigated the consequences of eliciting DNA damage in these embryos. We show that the rapid cell divisions are not affected by DNA damage. However, as the embryos reach the onset of gastrulation, they undergo rapid and synchronous apoptosis. We have investigated the regulation on this delayed apoptotic response to DNA damage. Next, we have reconstituted a DNA damage cell cycle checkpoint in vitro, demonstrating that all the checkpoint signalling components are present in the embryos but are not activated under the experimental conditions used to generate DNA damage in the embryo.

Reconstitution of an ATM-dependent checkpoint that inhibits chromosomal DNA replication following DNA damage.

Cell cycle checkpoints lead to the inhibition of cell cycle progression following DNA damage. A cell-free system derived from Xenopus eggs has been established that reconstitutes the checkpoint pathway inhibiting DNA replication initiation. DNA containing double-strand breaks inhibits replication initiation in a dose-dependent manner. Upon checkpoint activation, a prereplicative complex is assembled that contains ORC, Cdc6, Cdc7, and MCM proteins but lacks Cdc45. The checkpoint is ATM dependent. Cdk2/CyclinE acts downstream of ATM and is downregulated by Cdk2 phosphorylation on tyrosine 15. Cdk2AF/CyclinE is refractory to checkpoint signaling, and Cdc25A overrides the checkpoint and restores DNA replication. This report provides the description of a DNA damage checkpoint pathway that prevents the onset of S phase independently of the transcriptional function of p53 in a vertebrate organism.

Multicenter study on TGPO autoantibody prevalence in various thyroid and non-thyroid diseases; relationships with thyroglobulin and thyroperoxidase autoantibody parameters.

OBJECTIVE: TGPO autoantibodies (aAbs) that bind simultaneously to thyroglobulin (Tg) and thyroperoxidase (TPO) are present in the serum of patients with autoimmune thyroid diseases (AITD) and have been found to differ from monospecific Tg and TPO aAbs. To obtain further insights on the prevalence defined as the rate of occurrence and significance of TGPO aAbs in a large population, we carried out a collaborative study involving 15 European teams. METHODS: Serum samples from 3122 patients with various thyroid and non-thyroid diseases and normal subjects were assayed using a novel TGPO aAb detection kit. This test was designed so that TGPO aAbs are trapped between the Tg-coated solid phase and the soluble TPO labeled with a radioiodinated monoclonal antibody. RESULTS: Only three out of the 220 normal subjects (prevalence of 1.4%) were found to have positive TGPO aAb levels, which were mainly observed in the patients with AITD: the group of patients suffering from Hashimoto's thyroiditis had a TGPO aAb prevalence of 40.5% (n=437 patients), those with Graves' disease, a prevalence of 34.6% (n=645) and those with post-partum thyroiditis, 16.0% (n=243). Among the non-AITD patients with positive TGPO aAb levels, the TGPO aAb prevalence ranged from 20.7% among those with thyroid cancer (n=246) to 0% among those with toxic thyroid nodules (n=47). Among the patients with non-thyroid diseases, the TGPO aAb prevalence ranged from 9.8% in the case of Biermer's pernicious anemia (n=78) to 0% in that of premature ovarian failure (n=44). It is worth noting that the groups showing the highest TGPO aAb prevalence also contained the patients with the highest TGPO aAb titers. Statistical comparisons between the TGPO aAb prevalences in the various groups showed that TGPO aAb could be used as a parameter to distinguish between the groups of Hashimoto's and Graves' patients and between the women with post-partum thyroiditis and the post-partum women with only Tg and/or TPO aAb established during early pregnancy. Unexpectedly, the correlations between TGPO aAbs and Tg and TPO aAbs were found to depend mainly on the assay kit used. CONCLUSION: High TGPO aAb titers are consistently associated with AITD but the reverse was not found to be true. TGPO aAbs are a potentially useful tool, however, for establishing Hashimoto's diagnosis, and would be worth testing in this respect with a view to using them for routine AITD investigations.

Protein kinase A is required for chromosomal DNA replication.

Passage through mitosis resets cells for a new round of chromosomal DNA replication [1]. In late mitosis, the pre-replication complex - which includes the origin recognition complex (ORC), Cdc6 and the minichromosome maintenance (MCM) proteins - binds chromatin as a pre-requisite for DNA replication. S-phase-promoting cyclin-dependent kinases (Cdks) and the kinase Dbf4-Cdc7 then act to initiate replication. Before the onset of replication Cdc6 dissociates from chromatin. S-phase and M-phase Cdks block the formation of a new pre-replication complex, preventing DNA over-replication during the S, G2 and M phases of the cell cycle [1]. The nuclear membrane also contributes to limit genome replication to once per cell cycle [2]. Thus, at the end of M phase, nuclear membrane breakdown and the collapse of Cdk activity reset cells for a new round of chromosomal replication. We showed previously that protein kinase A (PKA) activity oscillates during the cell cycle in Xenopus egg extracts, peaking in late mitosis. The oscillations are induced by the M-phase-promoting Cdk [3] [4]. Here, we found that PKA oscillation was required for the following phase of DNA replication. PKA activity was needed from mitosis exit to the formation of the nuclear envelope. PKA was not required for the assembly of ORC2, Cdc6 and MCM3 onto chromatin. Inhibition of PKA activity, however, blocked the release of Cdc6 from chromatin and subsequent DNA replication. These data suggest that PKA activation in late M phase is required for the following S phase.

[Peritoneal splenosis simulating a right retroperitoneal mass]. / Splénose péritonéale simulant une masse rétropéritonéale droite.

We report a case of an incidentally diagnosed apparently isolated splenosis nodule located in the hepatorenal peritoneal recess. According to imaging features, this mass was a non-renal retroperitoneal tumor. In spite of the isolated location, the history of splenic rupture should have led to a heat-damaged red blood cell scintiscan and would thus have avoided percutaneous biopsy.

[Food preferences in anorectic girls at the beginning of therapy]. / Etude des préférences alimentaires chez la jeune fille anorexique au début de la prise en charge thérapeutique.

To determine the eating habits of patients with anorexia nervosa, we compared a group of 21 anorectic girls satisfying DSM-III-R criteria (A: 19 +/- 6.6 years; 14.2 +/- 2 kg/m2) with 30 control girls (C: 19.8 +/- 3.3 years; 20.9 +/- 4.4 kg/m2). A standardised form listing 226 food items was used to assess their food preferences. For each food category except vegetables and fish, the medium rate of positive appreciation was lower in group A than in group C. However, a positive correlation was found between the two groups (except for dairy and diet products), showing no major distortion of taste in group A. Anorectic girls generally discarded the sweetest fruit and the fattest meats, but sometimes chose to eat high-calorie food, possibly because of its supposed nutritional value. Their dislike for so-called "light" products was also apparent. Moreover, no regressive tendency to childish choices was found in their eating habits. It is concluded that group A displayed a narrowed food field, but without distortions of taste.

[Diabetes insipidus disclosing metastasis of breast adenocarcinoma]. / Diabète insipide révélant une métastase d'un adénocarcinome mammaire.

We are reporting on a case of diabetes insipidus (DI) and anterior pituitary failure revealing a breast cancer metastasis. Ten years after being diagnosed with a unilateral breast cancer, the patient presented with asthenia, thirst, polyuria and nocturia improved by subcutaneous DDAVP. MRI revealed a thickened pituitary stalk. DI is uncommon, late and usually asymptomatic in breast cancer. The association with an anterior pituitary failure is even more rare. In our patient the metastasis is in the pituitary stalk and seems to be due to meningeal deposits. MRI appears to be the best procedure to perform, showing a thickening stalk. Extension to the pituitary gland is related to direct tumor invasion from adjacent structures rather than haematogenous spread.

[Uroflowmetry in the early diagnosis of postoperative urethral stenosis]. / L'uroflussometria nella diagnosi precoce delle stenosi uretrali postoperatorie.

Uroflowmetry is a widely used technique for evaluation of lower urinary tract "obstructions", and urethral strictures (US) are considered a typically late complication after endoscopic or open prostatectomy. The clinical experience of the Department of Urology of Catania University is reported. Uroflowmetry was included in routine checks of prostatic patients after operation (TUR or open surgery) and the tests were performed at 40 and 180 days post-operatively. This proved to be an effective and objective means for early diagnosis of urethral strictures.

Nutritional status and growth in children with chronic hepatitis B.

As reported for other chronic liver diseases, hepatitis B virus (HBV) chronic infection might result in malnutrition. In order to establish whether this disease could be responsible of malnutrition and hence influence growth, 75 children, chronically infected with HBV, have been followed up for 4 years. Thirty-one of them had chronic active hepatitis (CAH), 25 chronic persistent hepatitis (CPH), 14 chronic lobular hepatitis (CLH), and five cirrhosis (three active, two inactive). The nutritional status was evaluated every 12 months, with careful physical and laboratory examinations. General nutritional status was estimated according to Waterlow criteria (13, 14). At our first observation, 50 children were following a balanced diet with a caloric intake adequate for age and weight, whereas 25 were on a low-fat diet, begun in the belief of its therapeutic value. For seven patients of this second group, the caloric intake was below the daily requirement. The latter group showed a growth failure in weight when they were first seen at our center and gained weight when the dietary intake was normalized. However, no biochemical feature of malnutrition was observed in all the 75 children. At the end of the follow-up period, the nutritional status was satisfactory for all of them.