Bone marrow transplantation improves the outcome of Atm-deficient mice through the migration of ATM-competent cells.

Ataxia telangiectasia (A-T) is a highly pleiotropic disorder. Patients suffer from progressive neurodegeneration, severe bronchial complications, immunodeficiency, hypersensitivity to radiotherapy and elevated risk of malignancies. Leukemia and lymphoma, along with lung failure, are the main causes of morbidity and mortality in A-T patients. At present, no effective therapy for A-T exists. One promising therapeutic approach is bone marrow transplantation (BMT) that is already used as a curative therapy for other genomic instability syndromes. We used an established clinically relevant non-myeloablative host-conditioning regimen and transplanted green fluorescent protein (GFP)-expressing ataxia telangiectasia mutated (ATM)-competent bone marrow-derived cells (BMDCs) into Atm-deficient mice. GFP expression allowed tracking of the potential migration of the cells into the tissues of recipient animals. Donor BMDCs migrated into the bone marrow, blood, thymus, spleen and lung tissue of Atm-deficient mice showing an ATM-competent phenotype. BMT inhibited thymic lymphomas, normalized T-lymphocyte populations, improved weight gain and rearing activity of Atm-deficient mice. In contrast, no GFP(+) cells were found in the cerebellum or cerebrum, and we detected decreased size index in MRI imaging of the cerebellum in 8-month-old transplanted Atm-deficient mice in comparison to wild-type mice. The repopulation with ATM-competent BMDCs is associated with a prolonged lifespan and significantly improved the phenotype of Atm-deficient mice.

Ectropion, entropion, trichiasis.

Aim of this review was to describe ectropion, entropion and trichiasis and their therapy. These eyelid pathologies are characterised by common symptoms (redness, excessive tearing and irritation of the eye) and by altered balance of the anterior and posterior lamellae of the eyelids. They involve more frequently the inferior eyelid and the therapy is mainly surgical. Parasurgical therapy may play a role as a temporary measure.

Pathoanatomy of cerebellar degeneration in spinocerebellar ataxia type 2 (SCA2) and type 3 (SCA3).

The cerebellum is one of the well-known targets of the pathological processes underlying spinocerebellar ataxia type 2 (SCA2) and type 3 (SCA3). Despite its pivotal role for the clinical pictures of these polyglutamine ataxias, no pathoanatomical studies of serial tissue sections through the cerebellum have been performed in SCA2 and SCA3 so far. Detailed pathoanatomical data are an important prerequisite for the identification of the initial events of the underlying disease processes of SCA2 and SCA3 and the reconstruction of its spread through the brain. In the present study, we performed a pathoanatomical investigation of serial thick tissue sections through the cerebellum of clinically diagnosed and genetically confirmed SCA2 and SCA3 patients. This study demonstrates that the cerebellar Purkinje cell layer and all four deep cerebellar nuclei consistently undergo considerable neuronal loss in SCA2 and SCA3. These cerebellar findings contribute substantially to the pathogenesis of clinical symptoms (i.e., dysarthria, intention tremor, oculomotor dysfunctions) of SCA2 and SCA3 patients and may facilitate the identification of the initial pathological alterations of the pathological processes of SCA2 and SCA3 and reconstruction of its spread through the brain.

Assessment of the multiple components of the variability in the adenoma detection rate in sigmoidoscopy screening, and lessons for training.

BACKGROUND AND STUDY AIMS: The determinants of the observed variability of adenoma detection rate (ADR) in endoscopy screening have not yet been fully explained. PATIENTS AND METHODS: Between November 1999 and November 2006 13 764 people (7094 men, 6670 women; age range 55-64) underwent screening flexible sigmoidoscopy at five hospital endoscopy units in Turin. To study the determinants of the ADR for distal adenomas, accounting for patient, examiner, and hospital characteristics, we applied a multivariate multilevel regression model. RESULTS: Average ADRs for all adenomas and for advanced adenomas (size > or = 10 mm, villous component > 20 %, high grade dysplasia) were 13.5 % (range 5.2 %-25.0 %) and 6.4 % (3.1 %-10.7 %) for men, and 8.0 % (2.5 %-14.0 %) and 3.7 % (0.2 % - 7.4 %) for women. In multivariate analysis, increased ADR of advanced adenomas was associated with male gender (odds ratio [OR] 1.78, 95 %CI 1.49 - 2.11), self-report of one first-degree relative with colorectal cancer (CRC) (1.44, 1.11-1.86), or of recent-onset rectal bleeding (1.73, 1.24-2.40). Adjusting for these variables, a significantly lower ADR was found for endoscopists with either a lower rate of incomplete sigmoidoscopy (< 9 %; OR 0.59, 95 %CI 0.41-0.87) or a higher rate (> 12 %; 0.64, 0.45-0.91), or with low activity volume (< 85 sigmoidoscopies/year; 0.66, 0.50-0.86). Residual variability explained by the endoscopy center effect was about 1 % and statistically significant. CONCLUSIONS: Endoscopist performance in flexible sigmoidoscopy CRC screening is highly variable. Low volume of screening activity independently predicts lower ADR, suggesting that operators devoting more time to screening sigmoidoscopy may perform better. Variability among pathologists in adenoma classification might explain part of the residual variability across endoscopy units.

Accumulation of phosphorylated I kappaB alpha and activated IKK in nodes of Ranvier.

AIMS: Nuclear factor-kappaB (NF-kappaB) is an ubiquitously expressed transcription factor that modulates inducible gene transcription crucial for the regulation of immunity, inflammatory processes, and cell survival. In the mammalian nervous system, constitutive NF-kappaB activation is considered to promote neuronal cell survival by preventing apoptosis. Increasing evidence suggests a critical role for NF-kappaB activation in acute and chronic neurodegenerative diseases. Recently, a striking enrichment of phosphorylated I kappaB alpha (pI kappaB alpha) and activated I KappaB Kinase (IKK), two key components of the NF-kappaB activation pathway, was demonstrated in the axon initial segment (AIS) of neurons. As the AIS shares fundamental features with nodes of Ranvier (NR), we examined whether pI kappaB alpha and activated IKK are also enriched in NR. METHODS: Double-immunofluorescence labelling was performed with vibratome sections of the rodent central and peripheral nervous system. Sections were analysed using confocal laser scanning microscopy and preembedding electron microscopy. RESULTS: Here we report a remarkable accumulation of pI kappaB alpha and activated IKK in NR in the central and peripheral nervous system. Immunolabelling for both proteins extended from NR into the adjacent paranode. pI kappaB alpha predominantly accumulated within the cytoplasm and was associated with fasciculated microtubules. This association was confirmed by electron microscopy. By comparison, activated IKK preferentially clustered beneath the cytoplasmic membrane. CONCLUSION: In conclusion, the coincident accumulation of pI kappaB alpha and activated IKK in AIS and NR suggests that these specific axonal compartments contribute to neuronal NF-kappaB activation.

Use of marginal donors in kidney transplantation: our experience.

The use of elderly donors has been advocated to expand the organ donor pool because of increased needs and the organ shortage. The aim of this study was to analyze whether the use of elderly donors and marginal kidneys affected the outcome of renal transplantations. Herein we presented data on 126 kidney transplantations performed from January 1996 to September 2003 using 32 marginal donors (group A) and 94 ideal donors (group B). We analyzed the medical and surgical complications and the graft survivals at a median follow-up of 18 months. Medical and surgical complications occurred in 22% and 5% versus 7% and 4% in groups A and B, respectively. The mean cold ischemia time and the mean age were greater for patients undergoing kidney transplantations from marginal donors. No differences were observed in graft survival in groups A and B. In conclusion, our data suggested that with an appropriate strategy and a correct selection of patients, marginal kidneys can be safely used to decrease the gap between demand and supply.

PTFE mesh in renal allograft compartment syndrome.

We report a case of anuria in a 42-year-old female kidney transplant patient that occurred secondary to extrinsic compression from a large kidney being placed extraperitoneally in a small iliac fossa. Prompt reexploration in the immediate postoperative period resulted in salvage of the graft with restoration of kidney function. The abdominal wall was reconstructed using prosthetic mesh, which decreased the compartment pressure within the iliac fossa sufficiently to allow the renal vein patency and the kidney perfusion. We think that this tension-free surgical technique should be applied in those cases in which the retroperitoneal space is less than the size of the kidney to avoid renal allograft compartment syndrome or incisional hernia.

Low-dose irradiation prior to bone marrow transplantation results in ATM activation and increased lethality in Atm-deficient mice.

Ataxia telangiectasia is a genetic instability syndrome characterized by neurodegeneration, immunodeficiency, severe bronchial complications, hypersensitivity to radiotherapy and an elevated risk of malignancies. Repopulation with ATM-competent bone marrow-derived cells (BMDCs) significantly prolonged the lifespan and improved the phenotype of Atm-deficient mice. The aim of the present study was to promote BMDC engraftment after bone marrow transplantation using low-dose irradiation (IR) as a co-conditioning strategy. Atm-deficient mice were transplanted with green fluorescent protein-expressing, ATM-positive BMDCs using a clinically relevant non-myeloablative host-conditioning regimen together with TBI (0.2-2.0 Gy). IR significantly improved the engraftment of BMDCs into the bone marrow, blood, spleen and lung in a dose-dependent manner, but not into the cerebellum. However, with increasing doses, IR lethality increased even after low-dose IR. Analysis of the bronchoalveolar lavage fluid and lung histochemistry revealed a significant enhancement in the number of inflammatory cells and oxidative damage. A delay in the resolution of γ-H2AX-expression points to an insufficient double-strand break repair capacity following IR with 0.5 Gy in Atm-deficient splenocytes. Our results demonstrate that even low-dose IR results in ATM activation. In the absence of ATM, low-dose IR leads to increased inflammation, oxidative stress and lethality in the Atm-deficient mouse model.

Proliferating and differentiating Schwann cell cultures from embryonic chick sciatic nerve maintained for months in vitro without antimitotics or growth factors.

In general, the available methods for culturing Schwann cells require specific antibodies and/or the addition of antimitotics to suppress fibroblasts, plus various factors to support their growth. Moreover, the maximal culture period of Schwann cells normally is limited to a few weeks. Here, three easy novel methods to culture Schwann cells from embryonic chick sciatic nerve are presented, that require no growth factors or agents elevating intracellular cAMP. In contrast to the conventional antimitotic treatment with cytosine arabinoside, we use D-valine to suppress fibroblasts. Our modified medium C leads within a few days to highly enriched Schwann cell cultures (culture I). Passage into a serum-reduced medium D allows for differentiating longterm cultures (culture II). In cultures I and II, the rate of cell division is low. However, after passage into serum-containing SC-medium, proliferation increases within one week to high levels (culture III). Cultures II and III can be grown for several months, during which time spontaneous immortalization can occur. The high purity of the cultures of about 95% is assessed using glia-specific antibodies for S-100 antigen, HNK-1 epitope, glia fibrillary acidic protein (GFAP), galactocerebroside (Gal C) and 3A7. These culture procedures are easy to perform and are suitable for differentiation, proliferation and coculturing experiments.

The intralaminar nuclei assigned to the medial pain system and other components of this system are early and progressively affected by the Alzheimer's disease-related cytoskeletal pathology.

The intralaminar nuclei of the human thalamus are integrated into the ascending reticular activating system and into limbic, oculomotor and somatomotor loops. In addition, some of them also represent important components of the medial pain system. We examined the occurrence and severity of the Alzheimer's disease (AD)-related cytoskeletal pathology and beta-amyloidosis in the seven intralaminar nuclei (central lateral nucleus, CL; central medial nucleus, CEM; centromedian nucleus, CM; cucullar nucleus, CU; paracentral nucleus, PC; parafascicular nucleus, PF; subparafascicular nucleus, SPF) in 27 autopsy cases at different stages of the cortical neurofibrillary pathology (cortical NFT/NT-stages I-VI) and beta-amyloidosis (cortical phases 1-4). The CEM, CL, PF, and SPF are slightly affected at stage II (corresponding to preclinical AD). They are markedly involved at stages III and IV (i.e. incipient AD) and severely affected at stages V and VI (i.e. clinical AD). In the PC and CU, the cytoskeletal pathology is mild at stage III, marked at stage IV, and severe at stages V-VI, whereas the CM is only mildly affected at stages IV-VI. In all of the intralaminar nuclei, deposits of the protein beta-amyloid occur for the first time during the final phase of cortical beta-amyloidosis. Functionally, the cytoskeletal pathology encountered in the intralaminar nuclei may contribute to the memory and affective symptoms, attention deficits, and dysfunctions related to horizontal saccades and smooth pursuits seen in AD patients. Equally important, however, are the findings that the cytoskeletal pathology developing within the intralaminar nuclei assigned to the medial pain system (CEM, CL, CU, PC, PF) as well as within other components of this system begins already during the preclinical or incipient phases of AD. Given this fact, the question arises as to whether non-discriminative aspects mediated by the medial pain system could be employed to identify individuals in the very earliest stages of AD.

Hospitalization for peptic ulcer bleeding: evaluation of a risk scoring system in clinical practice.

BACKGROUND: Upper gastrointestinal tract haemorrhage is a common cause of hospitalization: resource utilization in management of peptic ulcer bleeding varies considerably with no apparent effect on patient outcome. Several risk score systems based on endoscopic and clinical data have been proposed and validated in order to aid patient management. AIM: To assess clinical reliability of a scoring system and to define guidelines to improve efficiency of patient management without reducing efficacy METHODS: We considered all patients admitted to our unit for bleeding peptic ulcer over a one-year period. Every patient had an early endoscopy (within 12 hours) and therapy according to the appearance of the ulcer defined by Forrest classification. All subjects were classified into low-, intermediate- and high-risk patients on basis of clinical and endoscopic features according to "Cedar Sinai Medical Center predictive index" which was applied retrospectively in first six months then perspectively for the last period using the results obtained from first semester. For each risk group, we compared Length of Hospital Stay number of blood units used in transfusion, rebleeding rate, need for surgery as well as mortality in the two periods, using Student t test. We correlated Length of Hospital Stay and every score parameter by applying analysis of variance to results over the one-year period. RESULTS: Study population consists of 91 patients. Recurrent bleeding was observed in only three entering the high-risk group, only one of whom needed surgery Overall mortality was 9.8% (9 patients, only one for rebleeding). Variance analysis showed that the only parameter of the "Cedar Sinai Medical Center predictive index" which correlated with Length of Hospital Stay was comorbidity (p < or =0.05). Comparing the two periods, a close application of the score in the last six months allowed Length of Hospital Stay to be reduced in low-risk patients (t test with p=0.004) resulting in early discharge of 33% of cases without affecting patient outcome. CONCLUSIONS: This study confirms the reliability of the "Cedar Sinai Medical Center predictive index" in clinical practice improving the strategy of applying economic resources. Longer Length of Hospital Stay of intermediate- and high-risk groups is influenced more by comorbidities than by endoscopic findings. Early discharge was possible in one third of low risk patients. An accurate evaluation clinical para meters on admission together with early endoscopy may achieve the goal of reducing costs with a correct patient management.

Cocaine alters the onset and maintenance of maternal behavior in lactating rats.

Though much attention has been devoted to the behavioral and physiological consequences of cocaine abuse in offspring, little is known regarding the effects on the maternal behavior of the cocaine-exposed dam. We examined whether cocaine affects the initiation (late pregnancy) and/or maintenance (postpartum [PP]) phases of full maternal behavior (FMB; retrieving, grouping, and crouching over six pups) in Sprague-Dawley female rats. In Experiment 1, cocaine (5.0 or 10.0 mg/kg) or saline was administered on PP day 5 or 6 and FMB scored. Both dosages significantly disrupted FMB, particularly crouching, though 10.0 mg/kg had a greater effect on FMB. Experiment 2 (using 10.0 mg/kg cocaine) examined specific elements of the disruption and found significant reductions in proportion of females engaging in FMB, as well as increases in the latencies to contact, retrieve, lick, group, and crouch over pups. In Experiment 3 osmotic pumps containing 20 mg cocaine/kg/day or saline were implanted SC in day 14 pregnant rats. FMB testing was performed on days 1-2 postpartum together with a T-maze pup-retrieval test on postpartum days 3-5. Cocaine disrupted FMB in the homecage, in general, rendering the females less attentive to young, but was without effect in the T-maze tests. Cocaine--perhaps owing to its purported dopaminergic activity--may operate through motivational mechanisms to disrupt FMB in the postpartum maintenance phase; and through effects on late pregnancy levels of prolactin (a hormone which stimulates FMB), to disrupt maternal behavior during the initiation phase.

Normalization of bronchial exocrine pathology after endobronchial treatment with ambroxol.

A clinical study was performed on nine adult in-patients suffering from chronic intercrisis bronchopneumopathy, to evaluate whether a local endobronchial treatment with ambroxol could be effective in stabilizing the secretory activity of bronchial glands modified by phlogistic injury. The patients underwent bronchoscopic examination and deep biopsy, before and after treatment with local medication of ambroxol every other day for 30 days. In addition to morphometric measurements, detailed histochemical examinations were performed on the secretion of mucous acinic and glandular excretory ducts. The trial proved that the treatment with ambroxol stimulates serous acini, restoring normal production of sulphomucins and also of mucous acini, increasing the metabolic activities of acid glycoproteins and proportionally curtailing the excessive synthesis of glycoproteins. Ambroxol can therefore be considered a mucoregulating drug.

[Cardiac malformations with septal defects. Apropos of a case of biloculate heart with transposition and aortic atresia]. / Le malformazioni cardiache con difetti settali. A proposito di un caso di cuore biloculato con trasposizione ed atresia aortica.

The Authors describe a case of uniatrial univentricular heart with a right transposition and atresia of the aorta. This case forms part of an autoptic series of 74 congenital cardiopathies with defects of the septal system: monometameric (52 cases), polymetameric (22 cases), and limited (66 cases) or broad DIA, DIV or FAP types (8 cases). Having outlined the correlations between the type of defect and the complex morphogenetic events, whose close integration leads to a continuous septal system, the Authors focus their attention on the common associations between septal defects and other cardio-vascular anomalies (66 cases). The analysis of findings supports hypothesis that many of these are due to alterations in the position and direction of individual septal buds are should be seen against the overall background of septal pathology. Under these circumstances the pathogenetic importance of alterations of the complex movements presented by the cardiac type during the course of septal development and, to an event greater extent, possible alterations of the embryonal hematic flow are underlined.

[Ventricular hypoplasia in congenital heart diseases]. / Le ipoplasie ventricolari nelle cardiopatie congenite.

Twenty-seven cases of autopsy showing hypoplasia of one of the ventricular cavities, prevalently the left, were selected from a series of 103 congenital cardiopathies (1979-1990). In 9 cases ventricular hypoplasia formed part of complex malformative syndromes with a well-known physiognomy: a further 18 cases showed a complete (9 cases) or incomplete (9 cases) hypoplastic heart syndrome. Subjects were prevalently female and the presence of other malformations indicating a genetic dysfunction was less evident than in other cardiopathies. The different pathogenetic hypotheses are discussed in the light of macro- and microscopic morphological factors, of which the most plausible is that involving an anomalous position or orientation of the musculo-membranous folds which give rise to the septation system of the various metamers of the cardiac tube.

Extended pathoanatomical studies point to a consistent affection of the thalamus in spinocerebellar ataxia type 2.

The involvement of the thalamus during the course of the currently known polyglutamine diseases is still a matter of debate. While it is well-known that this diencephalic nuclear complex undergoes neurodegeneration in some polyglutamine diseases such as Huntington's disease (HD), it has remained unclear whether and to what extent the thalamus is also involved in spinocerebellar ataxia type 2 (SCA2) patients. Encouraged by our recent post-mortem findings in one German SCA2 patient and the results of a recent nuclear magnetic resonance (NMR) study, we extended our pathoanatomical analysis to serial thick sections stained for lipofuscin granules and Nissl substance through the thalami of four additional German and Cuban SCA2 patients. According to this analysis the thalamus is consistently affected by the destructive process of SCA2. In particular, during our study we observed a consistent involvement of the lateral geniculate body, the lateral posterior, ventral anterior, ventral lateral, ventral posterior lateral, and ventral posterior medial thalamic nuclei as well as the extraterritorial reticular nucleus. In four of the SCA2 cases studied additional damage was seen in the inferior and lateral nuclei of the pulvinar, whereas in the minority of the patients a subset of the limbic nuclei of the thalamus (i.e. anterodorsal, anteroprincipal, laterodorsal, fasciculosus, mediodorsal, central lateral, central medial, cucullar, and paracentral nuclei, medial nucleus of the pulvinar) underwent neurodegeneration. These interindividual differences in the distribution pattern of thalamic neurodegeneration indicate that the thalamic nuclei differ in their proclivities to degenerate in SCA2 and may suggest that they become involved at different phases in the evolution of the underlying degenerative process.

[Hepato-biliary complications of estro-progestin therapy]. / Complicanze epato-biliari della terapia estro-progestinica.

A brief account of the most frequently observed side-effects of the protracted administration of oestroprogestinic substances is followed by a detailed explanation of the damage they may cause to the hepatobiliary apparatus: cholestatic icterus; cholesterolic biliary lithiasis; benign and malignant liver neoplasias. Reference is made to the liver diagnosis problems the practitioner may encounter when dealing with patients treated with these substances. Stress is laid on the need for close monitoring of tolerance and the possible effects of oestroprogestinic management on liver and bile function.