Search

Altered mitochondrial DNA (mtDNA) methylation has been detected in several human pathologies, although little attention has been given to neurodegenerative diseases. Recently, altered methylation levels of the mitochondrial displacement loop (D-loop) region, which regulates mtDNA replication, were observed in peripheral blood cells of Alzheimer’s disease and amyotrophic lateral sclerosis patients. However, nothing is yet known about D-loop region methylation levels in peripheral blood of Parkinson’s disease (PD) patients. In the current study, we investigated D-loop methylation levels and mtDNA copy number in peripheral blood of 30 PD patients and 30 age- and sex-matched control subjects. DNA methylation analyses have been performed by means of methylation-sensitive high-resolution melting (MS-HRM) and pyrosequencing techniques, while mtDNA copy number was analyzed by quantitative PCR. MS-HRM and pyrosequencing analyses provided very similar D-loop methylation levels in PD patients and control subjects, and no differences between the two groups have been observed. Treatment with L-dopa and duration of the disease had no effect on D-loop methylation levels in PD patients. Additionally, mtDNA copy number did not differ between PD patients and control subjects. Current results suggest that D-loop methylation levels are not altered in peripheral blood of PD patients nor influenced by dopaminergic treatment.

Background: Red blood cells (RBCs) contain the majority of α-synuclein (α-syn) in blood, representing an interesting model for studying the peripheral pathological alterations proved in neurodegeneration. Objective: The current study aimed to investigate the diagnostic value of total α-syn, amyloid-β (Aβ1–42), tau, and their heteroaggregates in RBCs of Lewy body dementia (LBD) and Alzheimer’s disease (AD) patients compared to healthy controls (HC). Methods: By the use of enzyme-linked immunosorbent assays, RBCs concentrations of total α-syn, Aβ1–42, tau, and their heteroaggregates (α-syn/Aβ1–42 and α-syn/tau) were measured in 27 individuals with LBD (Parkinson’s disease dementia, n = 17; dementia with Lewy bodies, n = 10), 51 individuals with AD (AD dementia, n = 37; prodromal AD, n = 14), and HC (n = 60). Results: The total α-syn and tau concentrations as well as α-syn/tau heterodimers were significantly lower in the LBD group and the AD group compared with HC, whereas α-syn/Aβ1–42 concentrations were significantly lower in the AD dementia group only. RBC α-syn/tau heterodimers had a higher diagnostic accuracy for differentiating patients with LBD versus HC (AUROC = 0.80). Conclusion: RBC α-syn heteromers may be useful for differentiating between neurodegenerative dementias (LBD and AD) and HC. In particular, RBC α-syn/tau heterodimers have demonstrated good diagnostic accuracy for differentiating LBD from HC. However, they are not consistently different between LBD and AD. Our findings also suggest that α-syn, Aβ1–42, and tau interact in vivo to promote the aggregation and accumulation of each other.

Many physiological processes in the human body follow a 24-h circadian rhythm controlled by the circadian clock system. Light, sensed by retina, is the predominant “zeitgeber” able to synchronize the circadian rhythms to the light-dark cycles. Circadian rhythm dysfunction and sleep disorders have been associated with aging and neurodegenerative diseases including mild cognitive impairment (MCI) and Alzheimer’s disease (AD). In the present study, we aimed at investigating the genetic variability of clock genes in AD patients compared to healthy controls from Italy. We also included a group of Italian centenarians, considered as super-controls in association studies given their extreme phenotype of successful aging. We analyzed the exon sequences of eighty-four genes related to circadian rhythms, and the most significant variants identified in this first discovery phase were further assessed in a larger independent cohort of AD patients by matrix assisted laser desorption/ionization-time of flight mass spectrometry. The results identified a significant association between the rs3027178 polymorphism in the PER1 circadian gene with AD, the G allele being protective for AD. Interestingly, rs3027178 showed similar genotypic frequencies among AD patients and centenarians. These results collectively underline the relevance of circadian dysfunction in the predisposition to AD and contribute to the discussion on the role of the relationship between the genetics of age-related diseases and of longevity.

Background: Circadian and sleep disturbances are associated with increased risk of mild cognitive impairment (MCI) and Alzheimer’s disease (AD). Wearable activity trackers could provide a new approach in diagnosis and prevention. Objective: To evaluate sleep and circadian rhythm parameters, through wearable activity trackers, in MCI and AD patients as compared to controls, focusing on sex dissimilarities. Methods: Based on minute level data from consumer wearable devices, we analyzed actigraphic sleep parameters by applying an electromedical type I registered algorithm, and the corresponding circadian variables in 158 subjects: 86 females and 72 males (42 AD, 28 MCI, and 88 controls). Moreover, we used a confusion-matrix chart method to assess accuracy, precision, sensitivity, and specificity of two decision-tree models based on actigraphic data in predicting disease or health status. Results: Wake after sleep onset (WASO) was higher (p

A plethora of complex misfolded protein combinations have been found in Alzheimer disease (AD) brains besides the classical pathological hallmarks. Recently, α-synuclein (α-syn) and its heterocomplexes with amyloid-β (Aβ) and tau have been suggested to be involved in the pathophysiological processes of neurodegenerative diseases. These pathological features are not limited to the brain, but can be also found in peripheral fluids. In this respect, red blood cells (RBCs) have been suggested as a good model to investigate the biochemical alterations of neurodegeneration. Our aim is to find whether RBC concentrations of α-syn and its heterocomplexes (i.e., α-syn/Aβ and α-syn/tau) were different in AD patients compared with healthy controls (HC). The levels of homo- and heteroaggregates of α-syn, Aβ and tau, were analyzed in a cohort of AD patients at early stage either with dementia or prodromal symptoms (N = 39) and age-matched healthy controls (N = 39). All AD patients received a biomarker-based diagnosis (low cerebrospinal fluid levels of Aβ peptide combined with high cerebrospinal fluid concentrations of total tau and/or phospho-tau proteins; alternatively, a positivity to cerebral amyloid-PET scan). Our results showed lower concentrations of α-syn and its heterocomplexes (i.e., α-syn/Aβ and α-syn/tau) in RBCs of AD patients with respect to HC. RBC α-syn/Aβ as well as RBC α-syn/tau heterodimers discriminated AD participants from HC with fair accuracy, whereas RBC α-syn concentrations differentiated poorly the two groups. Although additional investigations are required, these data suggest α-syn heteroaggregates in RBCs as potential tool in the diagnostic work-up of early AD diagnosis.

Hemispatial neglect is common after unilateral brain damage, particularly to perisylvian structures in the right-hemisphere (RH). In this disabling syndrome, behaviour and awareness are biased away from the contralesional side of space towards the ipsilesional side. Theoretical accounts of this in terms of hemispheric rivalry have speculated that the intact left-hemisphere (LH) may become hyper-excitable after a RH lesion, due to release of inhibition from the damaged hemisphere. We tested this directly using a novel twin-coil transcranial magnetic stimulation (TMS) approach to measure excitability within the intact LH of neglect patients. This involved applying a conditioning TMS pulse over left posterior parietal cortex (PPC), in order to test its effect on the amplitude of motor evoked potentials (MEPs) produced by a subsequent test pulse over left motor cortex (M1). Twelve RH stroke patients with neglect, an age-matched group of eight RH stroke patients without neglect, and 10 healthy controls were examined. We found that excitability of left PPC-M1 circuits was higher in neglect patients than the other groups, and related to the degree of neglect on clinical cancellation tests. A follow-up found that 1 Hz repetitive TMS over left PPC normalized this over-excitability, and also ameliorated visual neglect on an experimental measure with chimeric objects. Our results provide 'direct' evidence for pathological over-excitability of the LH in the neglect syndrome, as quantified by left PPC influences on left M1, with implications for possible treatment. © The Author (2008). Published by Oxford University Press on behalf of the Guarantors of Brain. All rights reserved.

Visual perceptual learning is widely assumed to reflect plastic changes occurring along the cerebro-cortical visual pathways, including at the earliest stages of processing, though increasing evidence indicates that higher-level brain areas are also involved. Here we addressed the possibility that the cerebellum plays an important role in visual perceptual learning. Within the realm of motor control, the cerebellum supports learning of new skills and recalibration of motor commands when movement execution is consistently perturbed (adaptation). Growing evidence indicates that the cerebellum is also involved in cognition and mediates forms of cognitive learning. Therefore, the obvious question arises whether the cerebellum might play a similar role in learning and adaptation within the perceptual domain. We explored a possible deficit in visual perceptual learning (and adaptation) in patients with cerebellar damage using variants of a novel motion extrapolation, psychophysical paradigm. Compared to their age- and gender-matched controls, patients with focal damage to the posterior (but not the anterior) cerebellum showed strongly diminished learning, in terms of both rate and amount of improvement over time. Consistent with a double-dissociation pattern, patients with focal damage to the anterior cerebellum instead showed more severe clinical motor deficits, indicative of a distinct role of the anterior cerebellum in the motor domain. The collected evidence demonstrates that a pure form of slow-incremental visual perceptual learning is crucially dependent on the intact cerebellum, bearing the notion that the human cerebellum acts as a learning device for motor, cognitive and perceptual functions. We interpret the deficit in terms of an inability to fine-tune predictive models of the incoming flow of visual perceptual input over time. Moreover, our results suggest a strong dissociation between the role of different portions of the cerebellum in motor versusnon-motor functions, with only the posterior lobe being responsible for learning in the perceptual domain. © 2014.

BACKGROUND: The neural mechanisms and the circuitry involved in levodopa-induced dyskinesia (LID) are still partially obscure. LID can be considered the consequence of an abnormal pattern or code of activity that originates and is conveyed from the basal ganglia to the thalamus and the cortical motor areas. However, not only striatothalamocortical motor circuits but also other interconnected pathways could be implicated in its pathogenesis. METHODS: In a series of experiments, we applied repetitive transcranial magnetic stimulation (rTMS) over the lateral cerebellum in a group of patients with advanced Parkinson disease, to investigate whether modulation of cerebellothalamocortical circuits by means of rTMS may result in a modification of a dyskinetic state induced by levodopa ingestion. RESULTS: We found that a single session of cerebellar continuous theta burst stimulation (cTBS) was capable of transiently reducing LID. In the same patients, we observed that cerebellar cTBS changed the profile of activation of intracortical circuits in the contralateral primary motor cortex. Cerebellar cTBS reduced short intracortical inhibition and increased long intracortical inhibition, inducing a cortical reorganization that is associated with a reduction of LID. Furthermore, in another experiment, we observed that a 2-week course of bilateral cerebellar cTBS induced persistent clinical beneficial effects, reducing peak-dose LID for up to 4 weeks after the end of the daily stimulation period. CONCLUSIONS: Our study demonstrates that cerebellar continuous theta burst stimulation has an antidyskinetic effect in Parkinson disease patients with levodopa-induced dyskinesia, possibly due to modulation of cerebellothalamocortical pathways. © 2009 AAN Enterprises, Inc.

Objective: To investigate the efficacy of curcumin oral supplementation (600 mg/day, Brainoil), a natural antioxidant compound, in Amyotrophic Lateral Sclerosis (ALS). Methods: Patients were randomized into two groups: Group A received placebo for 3 months, then Brainoil for the following 3 months, Group B took Brainoil for 6 months. The evaluations were conducted at basal (T0), after 3 months of double blinded Brainoil or placebo treatment (T1), and after the 3 month open-label phase (T2). Clinical evaluations and oxidative stress biomarkers, including oxidative protein products (AOPPs), ferric reducing ability (FRAP), total thiols (T-SH) and lactate, were evaluated, compared to a control group, during an incremental forearm exercise test. Results: Over the entire study Group B showed a stable score of the ALS-FRS-r which decreased in Group A (pT0 exercise lactate was lower compared to Group A (p Conclusion: Although further studies are needed to confirm these data, treatment with curcumin shows encouraging results indicating a slight slowdown in disease progression, improving aerobic metabolism and oxidative damage, this also contributing to deepen knowledge into the pathogenic mechanisms of ALS.

Background Migraine is a complex multifactorial, neurobiological disorder, whose pathogenesis is not fully understood, nor are the mechanisms associated with migraine transformation from episodic to chronic pattern. A possible role of impaired oxidative mitochondrial metabolism in migraine pathogenesis has been hypothesized, and increased levels of peripheral markers of oxidative stress have been reported in migraine patients, although the literature data are limited and heterogeneous. Objectives The aim of this cross-sectional study was to determine plasmatic levels of advanced oxidation protein products, ferric-reducing antioxidant power and total plasmatic thiol groups, all plasmatic markers related to oxidative stress, in a sample of chronic migraine patients and medication-overuse headache, compared to a control group of healthy subjects. Methods Thirty-three patients with a diagnosis of both chronic migraine and medication-overuse headache (International Classification of Headache Disorders,3rd edition, beta version) and 33 healthy, headache-free subjects were enrolled. Patients with comorbid/coexisting conditions were excluded, as well as patients in treatment with migraine preventive drugs. Plasmatic levels of advanced oxidation protein products, ferric-reducing antioxidant power, and total thiol groups were determined in migraine patients and controls; moreover, oxidative stress biomarkers were compared in migraine patients with triptan compared to non-steroidal anti-inflammatory drug overuse. Results The statistical analysis showed significantly lower levels of ferric-reducing antioxidant power and total plasmatic thiol groups, both expression of antioxidant power, in patients with chronic migraine and medication-overuse headache compared to controls (respectively, ferric antioxidant power median [interquartile range] 0.53 [0.22] vs 0.82 [0.11] mmol/L, P

Neurodegenerative disorders (NDs) are characterized by abnormal accumulation/misfolding of specific proteins, primarily α-synuclein (α-syn), β-amyloid1-42 (Aβ), and tau, in both brain and peripheral tissue. In addition to homo-oligomers, the role of α-syn interactions with Aβ or tau has gradually emerged. The altered protein accumulation has been related to both oxidative stress and physical activity; nevertheless, no correlation among the presence of peripheral α-syn hetero-aggregates, antioxidant capacity, and physical exercise has been discovered as of yet. Herein, the content of α-syn, Aβ, tau, and of their heterocomplexes was determined in red blood cells (RBCs) of healthy subjects (sedentary and athletes). Such parameters were related to the extent of the antioxidant capability (AOC), a key marker of oxidative stress in aging-related pathologies, and to physical exercise, which is known to play an important preventive role in NDs and to modulate oxidative stress. Tau content and plasma AOC toward hydroxyl radicals were both reduced in older or sedentary subjects; in contrast, α-syn and Aβ accumulated in elderly subjects and showed an inverse correlation with both hydroxyl AOC and the level of physical activity. For the first time, α-syn heterocomplexes with Aβ or tau were quantified and demonstrated to be inversely related to hydroxyl AOC. Furthermore, α-syn/Aβ aggregates were significantly reduced in athletes and inversely correlated with physical activity level, independent of age. The positive correlation between antioxidant capability/physical activity and reduced protein accumulation was confirmed by these data and suggested that peripheral α-syn heterocomplexes may represent new indicators of ND-related protein misfolding.

The basal ganglia have been associated with temporal processing in ranges of milliseconds and seconds. However, results from PD patient studies are elusive. Time perception in these patients has been tested with different approaches including repetitive movement tasks (i.e. finger tapping) and cognitive tasks (i.e. time reproduction), and both abnormal and normal performances have been reported for different time intervals. Furthermore, when PD patients were required to learn two target durations in the same session when they were off medication, they overestimated the short duration and underestimated the long duration in the seconds range. This pattern of temporal accuracy was described as a "migration effect" and was interpreted as a dysfunctional representation of memory for time (Malapani, C., Rakitin, B. C., Levy, R., Meck, W. H., Deweer, B., Dubois, B., et al. (1998). Coupled temporal memories in Parkinson's disease: A dopamine-related dysfunction. Journal of Cognitive Neuroscience, 10, 316-331). Here, we controlled the emergence of similar behaviour also during millisecond time processing in PD patients. A time reproduction task was employed in which subjects were required to estimate intervals in millisecond (500 ms) and few second (2000 ms) ranges. In the first experiment, these intervals were tested in the same session to verify whether the migration effect was present also between time intervals in different millisecond and few second ranges. In a second experiment, they were not intermingled but were tested in two separate sessions to verify whether abnormalities depended on a selective perceptual deficit of the time intervals tested (i.e. millisecond or second ranges). All experiments were performed in both off and on therapy conditions. Our results demonstrated that PD patients showed no deficits in time estimation for time intervals in either the millisecond or few second range when the different time intervals were tested in separate sessions. This negative finding was obtained in both on and off conditions. However, when the different ranges were tested in the same session, we found that PD patients were impaired selectively for time intervals in the seconds range. Our data seem to indicate that time processing in PD patients for time intervals spanning up to 2 s is unimpaired and that abnormalities in such temporal scale may emerge only when patients have to deal with different durations, when timing involves further cognitive processes such as memory and attention. © 2007 Elsevier Ltd. All rights reserved.

A number of evidences indicates oxidative stress as a relevant pathogenic factor in Alzheimer's disease (AD) and mild cognitive impairment (MCI). Considering its recognized major genetic risk factors in AD, apolipoprotein (APO E) has been investigated in several experimental settings regarding its role in the process of reactive oxygen species (ROS) generation. The aim of this work has been to evaluate possible relationships between APO E genotype and plasma levels of selected oxidative stress markers in both AD and MCI patients. APO E genotypes were determined using restriction enzyme analysis. Plasma levels of oxidative markers, advanced oxidation protein products, iron-reducing ability of plasma and, in MCI, activity of superoxide dismutases were evaluated using spectrophotometric analysis. We found, compared to controls, increased levels of oxidized proteins and decreased values of plasma-reducing capacity in both AD patients (p0.0001) and MCI patients (p0.001); the difference between AD and MCI patients was significant only for plasma-reducing capacity (p0.0001), the former showing the lowest values. Superoxide dismutase activity was reduced, although not at statistical level, in MCI compared with that in controls. E4 allele was statistically associated (p0.05) with AD patients. When comparing different APO E genotype subgroups, no difference was present, as far as advanced oxidation protein products and iron-reducing ability of plasma levels were concerned, between E4 and non-E4 carriers, in both AD and MCI; on the contrary, E4 carriers MCI patients showed significantly decreased (p0.05) superoxide dismutase activity with respect to non-E4 carriers. This study, in confirming the occurrence of oxidative stress in AD and MCI patients, shows how it can be related, at least for superoxide dismutase activity in MCI, to APO E4 allele risk factor.

The vast majority of Alzheimer’s disease (AD) are late-onset forms (LOAD) likely due to the contribution of genetic, environmental, and stochastic factors, superimposed on a physiologically age-related decline of neuronal functions. Increasing evidence indicates epigenetic modifications in LOAD brains, and many of the environmental factors associated with AD risk, such as heavy metals and dietary factors, are able to modify the epigenome. There is also indication that environmentally-induced early life modifications of the genome during embryogenesis and brain development could contribute to the development of the disease later in life. DNA methyltransferase 3b (DNMT3b) is an enzyme involved in de novo methylation of the genome during embryogenesis, expressed in progenitor cells during neurogenesis. In the present study we evaluated two functional DNMT3B promoter polymorphisms, namely -149 C>T (rs2424913) and - 579 G>T (rs1569686), as candidate LOAD risk factors. Our analysis of 376 Italian LOAD patients and 308 matched controls revealed no difference in allele frequencies between the case an the control group (OR = 1.10 (0.88-1.39) for rs2424913, and OR = 1.02 (0.81-1.28) for rs1569686). Also the genotype distributions of both polymorphisms were closely similar between groups, and no significant effect on disease age at onset was observed. Overall, present results do not support a major role for rs2424913 or rs1569686 in LOAD pathogenesis.

Amyotropic lateral sclerosis (ALS) is a fatal and progressive neurodegenerative disease causing the loss of motoneurons of the brain and the spinal cord. The etiology of ALS is still uncertain, but males are at increased risk for the disease than females. Several studies have suggested that motoneurons in ALS might be subjected to the double insult of increased DNA oxidative damage and deficiencies in DNA repair systems. Particularly, increased levels of 8-oxoguanine and impairments of the DNA base excision repair system have been observed in neurons of ALS patients. There is evidence that the Ser326Cys polymorphism of the human 8-oxoguanine DNA glycosylase 1 (hOGG1) gene is associated with a reduced DNA repair activity. To evaluate the role of the hOGG1 Ser326Cys polymorphism in sporadic ALS (sALS), we screened 136 patients and 129 matched controls. In the total population, we observed association between both the Cys326 allele (p = 0.02) and the combined Ser326Cys + Cys326Cys genotype (OR = 1.65, 95% CI = 1.06–2.88) and increased risk of disease. After stratification by gender, the Cys326 allele (p = 0.01), both the Ser326Cys genotype (OR = 2.14, 95% CI = 1.09–4.19) and the combined Ser326Cys + Cys326Cys genotype (OR = 2.15, 95% CI = 1.16–4.01) were associated with sALS risk only in males. No significant association between the Ser326Cys polymorphism and disease phenotype, including age and site of onset and disease progression, was observed. Present results suggest a possible involvement of the hOGG1 Ser326Cys polymorphism in sALS pathogenesis.

Oxidative damage accumulates in the DNA of the human brain over time, and is supposed to play a critical role in the pathogenesis of Alzheimer's disease (AD). It has been suggested that the brain in AD might be subjected to the double insult of increased oxidative stress, as well as deficiencies in repair mechanisms responsible for the removal of oxidized bases. The type of damage that is most likely to occur in neuronal cells is oxidative DNA damage which is primarily removed by the base excision repair (BER) pathway, and a decrease in BER activity was observed in post-mortem brain regions of AD individuals, especially in the activity of 8-oxoguanine DNA glycosylase. There is evidence that the Ser326Cys polymorphism of the human 8-oxoguanine DNA glycosylase 1 (hOGG1) gene is associated with a reduced DNA repair activity. However, although a deficient BER was proposed in the etiology of AD by several authors, polymorphisms of BER genes have not been studied in AD yet. We performed a case-control study including 178 patients with sporadic AD (sAD) and 146 matched controls to evaluate the role of the Ser326Cys polymorphism as a risk factor for sAD. In the present study we failed to find any association between allele (chi2=0.03, p=0.86) or genotype (chi2=0.25, p=0.882) frequencies of hOGG1 Ser326Cys and the risk of sAD. Present results suggest that the Ser326Cys polymorphism of the hOGG1 gene is not an independent risk factor for sAD.

Alzheimer's disease (AD) is characterized by altered functional cortico-cortical connectivity likely due to loss of afferent and efferent connections between different cortical areas. Here we explored parieto-frontal functional connectivity in 15 AD patients and 12 healthy control subjects by means of bifocal transcranial magnetic stimulation (TMS). Conditioning stimuli were applied over the right posterior parietal cortex (PPC) at different intensities (90% and 110% of resting motor threshold, RMT). Motor evoked potentials (MEPs) were then recorded from the ipsilateral primary motor cortex at different interstimulus intervals (ISIs) ranging between 2 and 15 ms. Results showed that in healthy subjects, a conditioning TMS pulse applied over the ipsilateral PPC at 90%, but not at 110%, of RMT intensity was able to increase the excitability of the right M1. This functional interaction peaked at ISI = 6 ms. Conversely, in AD patients the facilitatory pattern of parieto-motor connections was evident only when TMS was delivered at an intensity of 110% of RMT with a peak at ISI = 8 ms. Moreover in AD patients, treatment with cholinesterase inhibitors did not induce any significant modification in the strength of the connection. In subsequent analyses, we found that, in AD patients, the effects induced by PPC conditioning at 110% RMT correlated with neuropsychological measures of episodic memory and executive functions, implying that patients with better cognitive performance had less impaired connectivity. Our findings reveal that parieto-frontal cortico-cortical functional connectivity is altered in AD patients, providing further evidence for a disconnection-based interpretation of AD symptoms. © 2013 - IOS Press and the authors. All rights reserved

Patients affected by Parkinson's disease (PD) may present with lower urinary tract (LUT) dysfunction characterized by involuntary detrusor overactivity. We evaluated possible impact of a 2-week course of low frequency 1 Hz repetitive transcranial magnetic stimulation (rTMS) on LUT behavior in eight advanced PD patients complaining of urinary disturbances. We tested the effects of rTMS measuring urodynamic examination and the International Prostate Symptoms Score (IPSS) questionnaire, used for evaluation of subjective LUTS. rTMS was able to improve temporarily LUT behavior in PD patients, increasing bladder capacity and the first sensation of filling phase. Moreover, a reduction of IPSS score was noticed, due to an improvement on filling phase symptoms. The beneficial effects assessed with the IPSS lasted for up to 2 weeks after the end of the stimulation. rTMS seems to be an effective, noninvasive alternative treatment for PD patients with urinary disturbances. © 2009 Movement Disorder Society.

The aim of the present study was to investigate the possible contribution of three common functional polymorphisms in the DNA repair protein X-ray repair cross-complementing group 1 (XRCC1), namely Arg194Trp (rs1799782), Arg280His (rs25489) and Arg399Gln (rs25487), to sporadic amyotrophic lateral sclerosis (SALS). We genotyped 206 Italian SALS patients and 203 matched controls for XRCC1 Arg194Trp, Arg280His and Arg399Gln polymorphisms by means of PCR/RFLP technique, searching for association between any of the studied polymorphisms and disease risk, age and site of onset. We observed a statistically significant difference in XRCC1 Gln399 allele frequencies between SALS cases and controls (0.39/0.28; p=0.001). The present study suggests that the XRCC1 Arg399Gln polymorphism might contribute to SALS risk.

Quadrantectomy and associated sentinel lymph node biopsy (SLNB) is currently employed in most breast surgery centres as the gold standard in the treatment of early breast cancer. This approach has a modest morbidity and can usually be performed in a day-surgery regimen, leading to best acceptance by the patients. This reports outlines the experience of our Breast Unit with quadrantectomy and SLNB in day surgery for early breast cancer. One hundred patients presenting to our institution with primary invasive breast cancer measuring less than 3 cm and clinically negative axillary nodes underwent quadrantectomy and SLNB in day surgery. For 60 women with breast cancer the sentinel node was negative, so the only definitive surgical treatment was performed in the day-surgery regimen; 40 patients with positive sentinel nodes were hospitalised a second time for axillary dissection. In these patients that needed clearance of the axilla, SLNB was performed on the only positive node in 22 cases (55%). None of the patients admitted for quadrantectomy and SLNB in day surgery required re-hospitalisation after discharge. All patients proved to be fully satisfied with early discharge from hospital when questioned on the occasion of subsequent monitoring. Short-stay surgical programs in early invasive breast cancer treatment are feasible today owing to the availability of less invasive approaches such as quadrantectomy and SLNB. There are two main pointers to a distinct advantage for this kind of approach, i.e. recovery and psychological adjustment. Recovery from surgery is faster and the patient tends to play down the seriousness of the operation and to have a better mental attitude to neoplastic disease. Moreover, when performing quadrantectomy with SLNB in day surgery fewer than 50% of breast cancer patients (40% in our experience) require another surgical treatment, concluding the surgery in a single session.

Sentinel lymph-node biopsy is an innovative method for axillary staging in breast cancer patients, based on the concept that information about the status of the entire lymphatic drainage from a tumour site could be obtained by identification and sampling of a "sentinel node". The aim of the study was to evaluate the impact of sentinel lymph-node biopsy in the management of patients with early invasive breast carcinoma. Three hundred and forty-one patients with primary invasive breast carcinoma measuring less than 2 cm (less than 3 cm from January 2001) and clinically negative axillary nodes were recruited into the study. Sentinel lymph-nodes were positive for metastases in 108/341 cases (31.7%). Micrometastases were found in 22 patients and isolated tumour cells in 1 case. The mean number of sentinel lymph-nodes removed was 1.8 per patient. The sentinel lymph-node was the only positive node in 57 of 108 patients (52.8%). The percentage of axillary recurrence in sentinel lymph-node-negative patients was 0%. The accuracy of sentinel lymph-node biopsy for axillary staging has been confirmed in many studies. Axillary recurrences after sentinel lymph-node biopsy range from 0 to 1.6% in many series, while axillary recurrence after axillary lymph-node dissection is about 0-3%. In our experience we observed no axillary recurrences in 233 patients with sentinel lymph-node biopsy alone, with a median follow-up of 33 months, confirming the accuracy of the procedure, and sentinel lymph-node-negative patients with sentinel lymph-node biopsy alone are no more at risk for axillary recurrences than those undergoing axillary lymph-node dissection.

Impairments in DNA repair enzymes have been observed in amyotrophic lateral sclerosis (ALS) tissues, particularly in the activity of the apurinic/apyrimidinic endonuclease 1 (APEX1). Moreover, it was suggested that the common APEX1 Asp148Glu polymorphism might be associated with ALS risk. To further address this question we performed the present study aimed at evaluating the contribution of the APEX1 Asp148Glu polymorphism in sporadic ALS (sALS) risk and clinical presentation, including age and site of onset and disease progression. We screened 134 sALS Italian patients and 129 matched controls for the presence of the APEX1 Asp148Glu polymorphism. No difference in APEX1 Asp148Glu allele and genotype frequencies was found between the groups, nor was the polymorphism associated with age and site of onset or disease progression. Present results do not support a role for the APEX1 Asp148Glu polymorphism in sALS pathogenesis in the Italian population.

Life expectancy for patients with breast carcinoma has changed in Europe over the last two decades. In Italy, the overall survival rate is about 77% at 5 years. When considering the situation in Sicily, the EUROCARE 2 study examined survival data from the Ragusa Cancer Registry, showing that the curves are worse than in other regions of Italy. Starting from these considerations we decide to evaluate whether these data from the Ragusa Cancer Registry corresponded to Palermo data. So we analysed data from 575 consecutive patients with breast cancer, treated in our Breast Unit from 1990 to 2003 according to the St. Gallen Recommendations and followed for a median period of 5 years. The prognostic role of age, tumour size, nodal status, TNM, stage, grading and hormonal receptors (OR, PR) were analysed and survival curves at 5 and 10 years were produced using the actuarial survival methods. All causes of death were considered. The median follow-up was 33 months. The Log rank test and univariate cox proportional model were used to demonstrate the association between prognostic factors and outcome. When considering T and N status, the curves showed an inverse correlation between survival and increases in these parameters. Overall survival was 92.9% at 5 years and 81.4% at 10 years for T1, 78.4% at 5 years and 61.4% at 10 years for T2 and 40.8% for T3-T4 at 5 and 10 years. Overall survival for NO was 92.1% and 78.2%, respectively, at 5 and 10 years, but decreased to 72.0% and 59.9% at 5 and 10 years for N1. In N2 patients we found that only about 50% of patients were still alive at 5 and 10 years, while for N3 patients the figures were 57.2% and 40%, respectively.

The aim of this study was to prospectively evaluate the sensitivity, specificity and accuracy of fine-needle aspiration of thyroid nodules. From January 1978 to December 2003, 497 patients underwent fine-needle aspiration of thyroid nodules and then thyroidectomy. Fine-needle aspiration was performed with a fine 23-25 gauge and the aspirate was fixed and dyed with May-Grounwald-Giemsa method. The results of cytology were divided into four groups: positive for malignancy (77 patients), negative for malignancy (370 patients), suspect for malignancy (34 patients) and not diagnostic (16 patients). Suspect specimens also included follicular neoplasm and Hürthle cell neoplasm. Fine-needle aspiration results were compared with histopathologic analysis after thyroidectomy. There were 2 false-positive (0.5%) and 5 false-negative patients (1%). Sensitivity, specificity and accuracy were respectively 94.7%, 99% and 98.4%. This study confirms that fine-needle aspiration of thyroid nodules can be performed easily with high sensitivity, specificity and accuracy.

The aim of the study was to evaluate possible differences in accuracy between the radioactive tracing and vital staining method in the search for sentinel nodes in patients with breast cancer. From January 1999 to December 2000, 102 patients with T1 N0 breast carcinoma were recruited into the study for localization of sentinel nodes with vital blue dye staining and radioactive tracing and were then submitted to lumpectomy and axillary dissection. For the two methods, we estimated the percentage of sentinel nodes localized, the false-negative rate, the predictive negative and positive value and the accuracy. The vital blue dye staining method permitted localization of the sentinel node in 73% of patients with a false-negative rate of 8%, a predictive negative value of 92% and 92% accuracy. The radioactive tracing method permitted localization of the sentinel node in 97% cases with a false-negative rate of 0%, a predictive negative value of 100% and 100% accuracy (P0.0005). The method that offers the better results is radioactive tracing. Currently, many authors use both techniques, since, in common practice, staining helps to identify the sentinel node with the probe.

Forty consecutive patients undergoing thyroid surgery under local anesthesia (LA) by a single surgeon over a 5-year period were included in this retrospective review. In all cases, the indication for LA was patient request. The study included 29 females and 11 males with an average age of 44 years (range 22-66 years). Body habitus was thin in 12.5%, average in 67.5%, and obese in 20%. Operations consisted of 21 unilateral thyroid lobectomies, 3 partial thyroidectomies, 3 subtotal thyroidectomies, and 13 total thyroidectomies. The pathology revealed benign disease in 45% and malignant disease in 55%. All procedures were performed using lidocaine and/or bupivacaine to administer a deep cervical plexus block as well as a field block. Mild additional intraoperative intravenous sedation was provided in most cases. Two patients were converted emergently to general endotracheal anesthesia because of inability to tolerate LA in one and a seizure secondary to intraarterial injection of lidocaine in the other patient. There were no instances of wound infection hemorrhage, recurrent laryngeal nerve injury, or hypoparathyroidism. In conclusion, thyroid surgery in selected patients can be performed safely using LA by experienced surgeons. If patients are carefully prepared preoperatively, LA offers a simple and reasonable alternative to general anesthesia.

The increase in the geriatric population in recent decades was not followed by a corresponding increase in attention on the part of surgeons to elderly patients until, in the '80s, many clinical studies showed that postoperative morbidity and mortality were related to the associated diseases rather than to the patients' age. To evaluate whether greater attention to the elderly is capable of increasing the number of operations that can be performed when necessary, we retrospectively surveyed the activity of a division of general surgery over two different periods of 5 years: the first one from 01/01/1985 to 31/12/1989 and the second from 01/01/1995 to 31/12/1999, when a team was specifically set up to take care of elderly patients. We registered a significant difference in the number of elderly patients operated on for tumours (134 vs. 208), an increase in colorectal neoplasms (24 vs. 70) and significant differences between radical and palliative surgery (P = 0.03). The extension of radical interventions for cancer to the elderly resulted in a moderate increase in postoperative complications. The surgery-related mortality was around 3.5% in both periods. Thus, a team of surgeons specifically dedicated to aged patients can improve the number of surgical operations performed in these subjects when required without any rise in postoperative mortality.

Background: Bilateral neck exploration (BNE) with the patient under general anesthesia has been the standard for parathyroidectomy. In efforts to minimize invasiveness and recovery from the procedure, unilateral neck exploration with the patient under local anesthesia in combination with sestamibi scanning is being done. Patients with a nonlocalized adenoma, concurrent thyroid disease, and/or multiple parathyroid adenomas have been excluded from this minimally invasive procedure. Methods: Two hundred thirty-six patients underwent BNE under local anesthesia for primary hyperparathyroidism that was performed by a single surgeon between 1988 and April 1999. Results: The percentage of patients who underwent parathyroidectomy under local anesthesia increased from 3% in 1988 to 97% in 1999. Twenty-three percent of patients underwent a concurrent thyroid procedure, and 84% of patients had a single adenoma removed. Sixty-two percent of patients had a negative preoperative sestamibi scan or did not have a scan at all. The incidence of frozen section decreased in the initial 50 cases from 100% to 39% during the last 100 cases. Average operative time was 43 minutes without a thyroid procedure and 66 minutes with a thyroid procedure. Overall, 70% of patients were discharged within 6 hours of the surgical procedure; this percentage increased to 91% during the last 5 months. Conclusions: BNE with the patient under local anesthesia can be performed safely and effectively in patients with coexisting thyroid disease and a nonlocalized adenoma. (Surgery 1999:126:1011-5.)

Background: The purpose of this paper was to review my evolving experience with local/regional anesthesia in an outpatient setting. Methods: Two hundred three consecutive patients during a 9-year period who chose to undergo thyroid operation under regional/local anesthesia were reviewed. Early discharge was offered to patients who were observed for 6 hours without neck swelling and who had no surgical reasons for delaying discharge. Results: In group A there were 2 patients who were given inhalation anesthesia during operation compared with none in groups B and C. The average length of stay in group A was 0.49 days, 0.55 days in group B, and 0.24 days in group C. Eighty-five percent of the patients whose operation began before 1300 hours were discharged within 6 hours versus only 50% of those operated on later in the day. Forty-seven percent of patients in group A, 65% of group B, and 77% of patients in group C were discharged within 6 hours of operation. On the basis of previous experience with general anesthesia, discharge time is not significantly influenced by the type of anesthesia chosen. There were no readmissions to the hospital, but 2 episodes of postoperative bleeding required reoperation. Survey showed that 95% of patients rated the level of pain equivalent or less severe than dental procedures under local anesthesia, and all patients would choose local again. Conclusions: These data suggest that thyroidectomy can be performed with the patient under local/regional anesthesia, with low morbidity and high patient satisfaction. Most patients can be discharged within 6 to 8 hours, and these discharges were not associated with readmissions. (Surgery 1998;124:975-9.)

Evidence suggests that atherosclerotic plaque composition and morphology are more important predictors of plaque stability and clinical behavior than the degree of vessel stenosis. Unfortunately, noninvasive methods of plaque characterization in situ using duplex ultrasound have been largely unsuccessful. We investigated the accuracy of helical computed tomography (HCT) in defining plaque morphology in patients undergoing carotid endarterectomy.Candidates for endarterectomy were enrolled prospectively and underwent HCT with intravenous contrast injection. The attenuation values (in Hounsfield Units) of plaque features such as calcium, fibrous tissue, and lipid were measured and correlated with the tissue types seen on microscopic examination. Percent luminal stenosis derived from transverse HCT images was also compared to values obtained from duplex ultrasound.Ambulatory patients referred to a tertiary, metropolitan medical center.A total of 21 out of 22 cases were suitable for comparison. Qualitatively, HCT accurately defined plaque features containing calcium, fibrous stroma, and lipid. Using tissue attenuation values (Hounsfield Units - H), HCT distinguished between lipid and fibrous stroma (means 39+/-12 H and 90+/-24 H, respectively, p0.001). HCT-derived calculations of percent stenosis also correlated with the ultrasound data (p0.001).HCT is an accurate, noninvasive means for studying detailed plaque morphology and composition. This technique provides the basis for further studies correlating plaque anatomy with clinical behavior.

Although there have been several reports in the literature describing a renewed interest in performing thyroid surgery under local anesthesia (LA), there has been little information regarding parathyroid surgery under local anesthesia.We retrospectively reviewed our experience of 49 LA parathyroid patients over a 9-year period at a single institution. A bilateral cervical block (C2-C3) was administered by a single surgeon using lidocaine and bupivacaine.The study included 39 females and 10 males with an average age of 62 years (range, 35-89 years). Every surgery was curative and the final pathology revealed 46 parathyroid adenomas and 3 cases of parathyroid hyperplasia. Forty-seven percent of the patients were discharged within 6 hours of operation and the remaining patients had a 1.4-day average length of hospital stay. A group of age- and sex-matched controls who underwent parathyroid surgery using general anesthesia (GA) served as a control group with 27% of operations performed as outpatients and an average length of stay of 1.6 days. Return to work averaged 6 days for the LA group versus 8 days for the GA. In the LA group, there was one instance of postoperative hemorrhage requiring reoperation and one instance of conversion to GA secondary to an inability to tolerate LA. There were no instances of recurrent laryngeal nerve injury or permanent hypoparathyroidism in either group.These data suggest that experienced surgeons can perform parathyroid surgery safely and effectively using LA as an alternative to GA.

A retrospective review of 500 patients with primary hyperparathyroidism seen from 1951 to 1975 was conducted; the effect of routine screening of calcium and phosphate levels (initiated in 1968) on the incidence and spectrum of the disease was analyzed. The majority of the patients (77%) were diagnosed in the eight-year period after routine biochemical screening was instituted. Comparing the group of patients diagnosed before the advent of biochemical screening and those diagnosed since screening was instituted, we found: (1) a small but significant increase in the number of asymptomatic patients diagnosed (from 2% to 12%); (2) no change in the incidence of related medical disorders, i.e., nephrocalcinosis and hypertension; (3) no change in the incidence of primary hyperplasia and adenoma; and (4) no change in the mean serum calcium level, the mean age at diagnosis, or the number or location of the involved parathyroid glands. Although routine calcium screening has identified significantly more cases of primary hyperparathyroidism, screening apparently does not enable diagnosis at an earlier stage.

Experimental studies disagree on the frequency and effects of reminiscence among the elderly. However, efficacy in working with recollection can be increased by an understanding of three types of reminiscence: informative, evaluative and obsessive. Informative reminiscence involves recollection for the pleasure of reliving and retelling. Reviewing memories can be used to revive interest, self-esteem, and personal relationships. Evaluative reminiscence is based on Butler's life review. A more favorable resolution is likely when it ranges over the lifetime and is shared. Groups and oral history projects can be tailored to support informative and evaluative reminiscences. Obsessive reminiscence may result from guilt, stress or grief. Relieving stress and finding new outlets can be combined with techniques like psychodrama and Progoff's journal workshop, to work through obstructing experiences.

Thyroglobulin (TG) is a glycoprotein which has been found to be produced by almost all nonmedullary thyroid cancers, and is present in elevated levels in the serum of most patients with metastatic thyroid cancer. It has long been a clinical impression that many metastatic thyroid cancers are "responsive" to TSH suppression as manifested by a decrease in the size and/or symptoms of the tumor. Five athyroid patients with metastatic thyroid cancer were studied to determine the effect of TSH stimulation on their serum TG levels and attempt to correlate this effect with their clinical response. Three of the five patients showed a definite rise in their serum TG once serum thyroxine (T4Y replacement was withdrawn, which correlated with an increase in TSH, and two of these patients have had no progression of their disease while on T4 suppression. The remaining two patients have shown no change in their TG levels after T4 withdrawal, and both have had progression of their metastatic disease while on T4 suppression. Thus, we see that in some patients, TG production by their metastatic thyroid cancer is responsive to TSH suppression, and there is some suggestion that TSH-responsive cancers are less aggressive; but it will require further studies to determine this.

Many thyroid malignancies are sufficiently differentiated to produce thyroglobulin both in situ and in perpipheral blood. Since patients who have undergone total thyroidectomy for malegnancy should not have normally circulating thyroglobulin, their serum thyroglobulin may provide a simple and specific tumour marker for recurrent disease. Of 30 such athyroid patients who were studied, all of the 20 patients who were disease-free ten years after thyroidectomy had minimal (less than15 ng/ml) serum-thyroglobulin levels while all of 10 patients with recurrences had raised levels (greater than 90 ng/ml). Controls ranged from 0 to 60 ng/ml. This assay should prove valuable in following patients who have undergone total thyroidectomy for recurrent thyroid malignancy.

Seven patients with advanced cutaneous T-cell lymphomas were treated with a combination chemotherapy regimen of vincristine, doxorubicin (Adriamycin), bleomycin, methotrexate, leucovorin factor, 5-FU, and hydrocortisone. All patients had an objective response but only one had a complete clinical remission. Our results are compared to other regimens reported in the literature.