Amyotrophic lateral sclerosis (ALS) is a neurodegenerative disorder consisting of progressive loss of motor neurons. TDP-43 has been identified as a component of ubiquitinimmunoreactive inclusions of motor neurons in ALS. We focused on the diffuse cytoplasmic TDP-43 immunoreactivity in ALS neurons, and quantitatively assessed it in comparison with skein/round TDP-43 and ubiquitin immunostaining in motor neurons of 30 sporadic ALS cases. The percentage of spinal motor neurons with cytoplasmic TDP-43 immunoreactivity was higher than that of ubiquitin-immunoreactive ones. The percentage of TDP-43-positive motor neurons was independent of neuron counts in anterior horns, while the percentage of ubiquitinated neurons was inversely correlated. Aiming to define the cytosolic localization of TDP-43, the immunoblot analysis of spinal cord and frontal cortex showed that full-length TDP-43, the 45 kDa form and ubiquitinated TDP-43 are found in the soluble inclusion-free fraction. The present data suggest that delocalization, accumulation and ubiquitination of TDP-43 in the cytoplasm of motor neurons are early dysfunctions in the cascade of the events leading to motor neuron degeneration in ALS, preceding the formation of insoluble inclusion bodies. Being cytoplasmic accumulation an ongoing event during the course of the illness, a therapeutic approach to this incurable disease can be envisaged.

The possible relationship between Guillain-Barré syndrome (GBS) and cancer is still controversial and the existence of a paraneoplastic GBS remains unconfirmed. To better define whether there is a relationship between GBS and malignancy, we compared the observed and the expected number of patients with tumours in a population-based cohort of subjects with GBS. Clinical differences between GBS patients with or without malignancies were analysed. Data were obtained from the Piemonte and Valle d'Aosta Register for GBS (PARGBS) (years 1990-1998). GBS was diagnosed according to NINCDS criteria. The number of expected cases of malignancy in the PARGBS population was calculated using the incidence rate of all types of cancer (ICD codes 140-208) in Piemonte [1985-1987], and in the most important town of this region, that is Turin (years 1993-1997). In the nine-year period, 435 incident patients with GBS were found. Nine of them developed cancer in the six months preceding or following GBS; in seven of them, the diagnosis of cancer and GBS was concomitant. The expected number of malignant tumours was 3.7 (using the incidence in Piemonte) and 3.8 (using the incidence in Turin); therefore, the odds ratios were 2.43 (95 % CI, 1.11-4.62) and 2.37 (95% CI, 1.09-4.50), respectively (p<0.01). Although the cases with malignancies were clinically similar to the other cases of GBS observed through the Register, the mortality in GBS patients with cancer was higher and was the final cause of death in two patients affected by severe cancer. These results suggest a possible correlation between some cases of GBS and cancer. However, GBS in cancer patients does not meet all the criteria for paraneoplastic diseases.

1Promoter methylation of the O6-methylguanine-DNA methyltransferase (MGMT) gene plays 2 a role in cellular response to alkylating agents. In the present study aimed to: i) evaluate the 3 concordance between MGMT promoter methylation status in tumor tissue and plasma; ii) 4 monitor MGMT promoter methylation status in plasma taken before and during 5 temozolomide treatment; iii) explore the value of MGMT promoter methylation status in 6 plasma as a prognostic/predictive biomarker in glioma patients. 7We enrolled 58 patients with histologically confirmed glioma at different grades of 8 malignancy. All patients underwent surgical resection and temozolomide treatment. Paraffin-9 embedded tumor tissue was available for 48 patients. Blood samples were collected from all 10 patients before temozolomide treatment (baseline) and at each MRI examination for a 12-11 month period. MGMT promoter methylation status was assessed in both sample types by real 12 time PCR with a specific probe. 13The frequency of MGMT promoter methylation was 60.4% in tumor tissue and 41.38% in 14 plasma. MGMT promoter methylation status was concordant in the two sample types 15 (Kappa=0.75, 95% confidence interval [CI] 0.57-0.93; p-value <0.001). Overall and 16 progression-free survival were longer in patients with methylated MGMT promoter. 17Mortality was higher in patients with unmethylated MGMT promoter, whether in tumor 18 tissue (hazard ratio [HR]: 2.21; 95% CI 0.99-4.95) or plasma (HR: 2.19; 95% CI 1.02-4.68). 19Progression-free survival was shorter in patients with unmethylated MGMT promoter, 20 whether in tissue (HR: 2.30; 95% CI 1.19-4.45) or plasma (HR: 1.77; 95% CI 0.95-3.30). 21The cumulative incidence of unmethylated MGMT promoter in plasma at baseline was 58%, 22 and reached virtually 100% at 12 months. In conclusion MGMT promoter methylation status 23 in tumor tissue and plasma was highly concordant, and both were associated with longer 24 survival, supporting the role of the detection of methylated MGMT promoter in predicting 25 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 3 treatment response. However we suggest caution in using plasma as a surrogate of tumor 1 tissue due to possible false-negative results. 2 3 4 Keywords: Glioma, plasma DNA, MGMT, methylation, temozolomide 5 6 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 16 17 18 19 20 21 22 23 24 25 26 27 28 29 30 31 32 33 34 35 36 37 38 39 40 41 42 43 44 45 46 47 48 49 50 51 52 53 54 55 56 57 58 59 60 61 62 63 64 65 4 Introduction 1 Gliomas are the most common type of primary brain tumor in adults, with a standardized 2 incidence in Europe of 6/100,000 person-years, and a peak of 18.5/100,000 person-years in 3 people over 65 years of age [1]. Despite recent advances in the treatment of gliomas, such as 4 ...

TMZ has a role in recurrent chemo-naïve adult patients with intracranial ependymoma, regardless of tumor grade and MGMT methylation. We suggest that, after failure of surgery and radiotherapy, TMZ should be considered as a possible first-line treatment for recurrent ependymoma.

Although a large number of amyotrophic lateral sclerosis (ALS) patients have undergone transplantation procedures with olfactory ensheathing cells (OECs) in the Bejing Hospital, to our knowledge, no post-mortem neuropathologic analyses have been performed. We examined the post-mortem brain of two Italian patients affected by ALS who underwent cellular transplantation in Beijing with their consent. Our aim was to assess the events following the graft procedure to possibly support the rationale of the treatment strategy. The neuropathologic findings were analyzed on the basis of the limited awareness of the experimental conditions and discussed in relation to the safety, efficacy and long-term outcome of the transplanted cells. Islands of quiescent, undifferentiated cells within the delivery track persisting for up to 12 months-24 months were found. Prominent glial and inflammatory reaction around the delivery track strongly supports the encasement of the graft. Evidence of axonal regeneration, neuronal differentiation and myelination was not seen. The surgical procedure of implantation was not compatible with a neurotrophic effect. The OEC transplantation did not modify the neuropathology of ALS in the two patients. In conclusion, the present neuropathologic analysis does not support a beneficial effect of fetal OEC implantation into the frontal lobes of ALS patients.