Spatial clustering of amyotrophic lateral sclerosis in Sardinia, Italy: The contribution of age, sex, and genetic factors.

INTRODUCTION/AIMS: Several microgeographic clusters of higher/lower incidence of amyotrophic lateral sclerosis (ALS) have been identified worldwide. Differences in the distribution of local factors were proposed to explain the excess ALS risk, whereas the contribution of known genetic/epigenetic factors remains unclear. The aim is to identify restricted areas of higher risk in Sardinia and to assess whether age, sex, and the most common causative genetic mutations in Sardinia (C9orf72 and TARDBP mutations) contributed to the variation in the ALS risk. METHODS: We performed an ad hoc analysis of the 10-y population-based incident cohort of ALS cases from a recent study of a large Sardinian area. Cluster analysis was performed by age- and sex-adjusted Kulldorff's spatial scan statistic. RESULTS: We identified a statistically significant cluster of higher ALS incidence in a relatively large area including 34 municipalities and >100,000 individuals. The investigated genetic mutations were more frequent in the cluster area than outside. Regardless of the genetic mutations, the excess of ALS risk was significantly associated with either sex or with age ≥ 65 y. Finally, an additive interaction between older age and male sex contributed to the excess of ALS risk in the cluster area but not outside. DISCUSSION: Our analysis demonstrated that known genetic factors, age, and sex may contribute to microgeographic variation in ALS incidence. The significant additive interaction between older age and male sex we found in the high-incidence cluster could suggest the presence of a third factor connecting the analyzed risk factors.

Increasing prevalence 2015-2019 of amyotrophic lateral sclerosis in Sardinia, Italy.

BACKGROUND: While amyotrophic lateral sclerosis (ALS) incidence has increased during the last decades, structured evidence on increased prevalence is lacking. After reporting a significant yearly increase of ALS incidence over a 10-year period, we checked for increased prevalence in Southern Sardinia over a quinquennium. METHODS: ALS patients (El Escorial Criteria) recruited from the study area and followed at ALS Centre, University of Cagliari, were included. Prevalence was computed for January 1, 2015 and January 1, 2019 and was calculated for the overall ALS population as well as for tracheostomized and non-tracheostomized patients. RESULTS: We observed a non-significant trend for greater ALS prevalence in 2019 than in 2015 (18.31 per 100,000 vs. 15.26 per 100,000; rate ratio: 1.83, p = 0.01). By contrast, a significantly raising 2015 to 2019 ALS prevalence was observed in tracheostomized patients. No significant difference could be detected in non-tracheostomized. CONCLUSIONS: We provided the highest prevalence rate to date reported in the worldwide literature, and also showed a non-significant raising ALS prevalence in the Sardinian population over a quinquennium. The trend in raising ALS prevalence was likely due to extended survival due to invasive interventions.

Disease survival and progression in TARDBP ALS patients from Sardinia, Italy.

BACKGROUND: Common genes implicated in amyotrophic lateral sclerosis (ALS) development may also influence its progression rate. The C9orf72 mutations featured a faster progression rate while the European SOD1 mutations were associated with a slower progression. In this study, we assessed the relationship between TARDBP and ALS progression/survival. METHODS: ALS incident patients (2010-2019) were diagnosed by El Escorial revised criteria and staged over the disease course by the King's staging system. Disease progression was analysed by Kaplan-Meier survival curves and Cox regression models, with survival measured from symptom onset to death/tracheostomy or censor date. RESULTS: The study population included 76 patients carrying TARDBP mutations (A382T/G295S), 28 patients carrying the C9orf72 GGGGCC expansion, and 158 patients who had no evidence of causative genetic mutations (nmALS group). TARDBP patients reached death/tracheostomy later than C9orf72 and nmALS patients, independently of possible prognostic indicators (sex, age at ALS onset, diagnostic delay, phenotype at onset, and family history of ALS). On King's staging, the time elapsed between disease onset (King's stage 1) and involvement of the second body region (King's stage 2B) was similar in TARDBP and nmALS patients but longer in TARDBP than in C9orf72 patients. TARDBP patients reached King's stages 3 and 4 later than C9orf72 and nmALS patients. CONCLUSIONS: TARDBP patients have a better survival/prognosis than C9orf72-positive and nmALS patients. King's staging also suggested that the higher survival rate and the slower progression associated with the TARDBP mutation could mainly be attributed to the longer time elapsed between King's stages 2B to 3.

Percutaneous gastrostomy, mechanical ventilation and survival in amyotrophic lateral sclerosis: an observational study in an incident cohort.

OBJECTIVE: To analyze disease-modifying effects of percutaneous endoscopic gastrostomy (PEG) insertion for supporting nutrition, noninvasive ventilation (NIV), and tracheostomy-assisted ('invasive') ventilation (TIV) in amyotrophic lateral sclerosis (ALS). METHODS: We retrospectively analyzed survival in a large population-based incident cohort that was prospectively followed up in our center. Analysis considered several known ALS-related prognostic variables. RESULTS: In this population, PEG and NIV in multivariable analysis significantly correlated to survival as computed by disease onset to death/tracheostomy. NIV was associated with better survival while PEG was associated with reduced survival. Other independent prognostic factors were age at ALS onset, diagnostic delay, and flail arm/leg and pure upper motor neuron (PUMN) phenotypes. The length of survival after TIV was significantly associated with age at ALS onset (inverse correlation) whereas other variables did not. The length of survival after TIV correlated to age at ALS onset in such a way that each additional year of age at ALS onset decreased survival by about 0.7 months. Patients who underwent both TIV and NIV did not experience a better survival than those who underwent TIV alone. CONCLUSION: The lack of effect of enteral nutrition on ALS survival probably reflected the timing of PEG insertion in patients with more severe disease. By contrast, patients who used mechanical ventilation had an increased overall survival compared with non-ventilated ones. The study also provided new information showing that the combined use of NIV and TIV did not may prolong ALS survival as compared to TIV alone.

Impact of occupational categories on the incidence of amyotrophic lateral sclerosis in Sardinia Island, Italy.

OBJECTIVE: Occupation is one of the potential risk factors for amyotrophic lateral sclerosis (ALS) for which previous controlled studies produced inconsistent results. The aim of this study is to assess the impact of several groups of occupational categories on ALS incidence. METHODS: ALS patients from the southern part of Sardinia who had onset during 2012-2021 and fulfilled El Escorial revised diagnostic criteria were included. The risk of ALS was estimated in relation to the occupation held in 2011, as obtained from the 2011 Census that classified working activities in ten groups. Each occupational group was compared with a reference category represented by all other occupations, and rate ratio were calculated. Additive interaction between activity at work and age at ALS onset/sex on ALS incidence was calculated. RESULTS: Employment in agriculture/breeding and in the armed forces were significantly associated with increasing ALS risk. None of the other assessed occupation groups was associated with change in the risk of ALS. Geographic analysis indicated that the effect of agriculture/breeding was particularly evident in the areas of higher risk for the general population. By contrast, an inverse pattern of spatial risk was associated with armed forces activity at work. The increased risk of ALS associated to agriculture/breeding was more evident in older people. No significant interaction was detected between working in the armed forces and older age/sex. CONCLUSIONS: The significant interaction between agriculture/breeding and age suggests that the mechanisms leading to ALS are complex and involve several factors.

Tyrosine kinase receptor status in endometrial stromal sarcoma: an immunohistochemical and genetic-molecular analysis.

Endometrial stromal sarcomas (ESS) are rare uterine malignant mesenchymal neoplasms, which are currently treated by surgery, as effective adjuvant therapies have not yet been established. Tyrosine kinase inhibitors have rarely been applied in ESS therapy, with few reports describing imatinib responsivity. The aim of this study was to analyze the status of different tyrosine kinase receptors in an ESS series, in order to evaluate their potential role as molecular targets. Immunohistochemistry was performed for EGFR, c-KIT, PDGFR-α, PDGFR-ß, and ABL on 28 ESS. EGFR, PDGFR-α, and PDGFR-ß gene expression was investigated by real-time polymerase chain reaction (qRT-PCR) on selected cases. "Hot-spot" mutations were screened for on EGFR, c-KIT, PDGFR-α, and PDGFR-ß genes, by sequencing. All analysis was executed from formalin-fixed, paraffin-embedded specimens. Immunohistochemical overexpression of 2 or more tyrosine kinase receptors was observed in 18 of 28 tumors (64%), whereas only 5 tumors were consistently negative. Gene expression profiles were concordant with immunohistochemical overexpression in only 1 tumor, which displayed both high mRNA levels and specific immunoreactivity for PDGFR-α, and PDGFR-ß. No activating mutations were found on the tumors included in the study. This study confirms that TKRs expression is frequently observed in ESS. Considering that the responsiveness to tyrosine kinase inhibitors is known to be related to the presence of specific activating mutations or gene over-expression, which are not detectable in ESS, TKRs immunohistochemical over-expression alone should not be considered as a reliable marker for targeted therapies in ESS. Specific post-translational abnormalities, responsible for activation of TKRs, should be further investigated.

Incidence of amyotrophic lateral sclerosis in Sardinia, Italy: age-sex interaction and spatial-temporal variability.

Objective: This study assessed amyotrophic lateral sclerosis (ALS) incidence in Sardinia, Italy, and the combined contribution of age and gender to disease risk. We also checked disease incidence for spatial-temporal variability. Methods: ALS patients from all neurological centers of the study area who had onset during 2010-2019 and fulfilled El Escorial revised diagnostic criteria were included. Incidence was calculated for the overall study area and each province separately. Additive interaction between age and sex on ALS incidence was assessed. Results: The average crude annual incidence rate was 3.6/100,000 person-years (95% CI, 3.2-4.1), 3.1/100,000 person-years (95% CI, 2.7-3.5) when age-adjusted. Incidence was greater among people aged ≥65 years and men, with the two variables undergoing significant additive interaction. Incidence increased yearly over the study period, with annual incidence correlating with the increasing yearly frequency of people aged ≥65 years, but not with the proportion of incident cases carrying genetic mutations. Stratifying by province, the rates from Oristano and South Sardinia were higher than the rate from Cagliari. ALS patients from areas at different risk were comparable for frequency of clinical/genetic features. Conclusion: ALS incidence in Sardinia was in the upper part of the European range of variability. We also provided new information about age and sex as risk factors for ALS, showing male sex as a modifier of the effect of aging on ALS incidence. Spatial-temporal variations in ALS incidence correlated to changes in the proportion of the aging population rather than to the distribution of genetic factors.

Black hairy tongue in a patient with amyotrophic lateral sclerosis.

Black hairy tongue (BHT) is a condition characterized by the elongation of filiform papillae associated with a marked discoloration, from yellowish-brown to black, and a thick lingual coating. BHT is usually observed in the elderly and in patients with limited self-sufficiency, as a consequence of poor oral hygiene. In this perspective, the patients affected by amyotrophic lateral sclerosis (ALS) represent a high-risk category for the occurrence of BHT. The fast and inexorable loss of their self-sufficiency due to progressive muscle atrophy as well as the impropriate education of healthcare assistants have demonstrated to have significant reflection on the maintenance of an adequate standard of oral hygiene. This paper firstly described a case of BHT in a patient affected by ALS. A case of BHT in a patient (Caucasic, male, 63 years old) affected by ALS was described. The primary goal of the work was to teach and motivate the patient to the use of the tongue cleaner in association with the local application of chlorexidine 0.20%. Furthermore, in order to support the patient with accurate domiciliary oral hygiene, a proper training for his health-care assistant was provided. The maintenance of the oral health of ALS patient is fundamental to prevent systemic complications that could jeopardize the already fragile physical balance of these patients. The dedicated monitoring by a dentist or a dental hygienist would seem essential in order to achieve this objective.

Probing depth in periodontal pockets: In vitro evaluation of contributions to variability due to probe type and operator skill.

Periodontal probing aims at detecting the presence, type and gravity of periodontal diseases influencing distance between gingival margin and connective ligament. Measurements in vivo are affected by substantial uncertainty, owing, for example, to probe features, anatomic variations and operator's skill. Inadequate reproducibility in periodontal probing may lead to diagnostic mistakes and inappropriate therapeutic decisions. In vitro evaluation of reproducibility of measurements of periodontal pockets was aimed at while developing a Periodontal Calibration Box designed to calibrate periodontal probe operators, catering also for simulating contraction observed in vivo. Probe type and clinical experience of operators were found to affect substantially probing errors.

Application of the novel method in the diagnosis and treatment of median rhomboid glossitis Candida-associated.

The aim of this work is to demonstrate how current molecular techniques should be integrated in the diagnostic process and can have a crucial role in the management of oral fungal infections. A case of median rhomboid glossitis Candida-associated and its resolution will be described step by step. At the time of the first observation, the lesion on the surface of the tongue did not respond to the previous administration of topical antifungal agent, such a nystatin. Firstly, in order to identify the causative agent and confirm Candida albicans infection, a brushing of the lesion was performed and polymerase chain reaction analysis was carried out. In addition, deoxyribonucleic acid sequencing method, known as Pyrosequencing(®), was used in the detection of point mutations commonly associated with fluconazole resistance and consequently, in the prediction of susceptibility to azole agents. According to molecular findings, the administration of fluconazole has therefore led to resolution of the case in 2 weeks. This case highlights how the use of molecular techniques, now-a-days, can assist the clinicians to quickly obtain the report with highly accurate and precise results and appropriately support them in the diagnosis and therapeutic process.

Microbial changes in subgingival plaque and polymicrobial intracellular flora in buccal cells after fixed orthodontic appliance therapy: a preliminary study.

The oral ecosystem is strictly related to a balance maintained by specific niches recognized as sites, where oral bacteria can metabolize avoiding the immune system response. The oral bacteria species that colonize the ecological niches vary during fixed orthodontic treatment, with a prevalence of periodontal bacterial species. Qualitative analysis of five periodontal pathogens was used to investigate the microbial colonization rate in the crevice and buccal epithelial cells. The presence of inadequate oral hygiene was considered as a modulation variable for microbial colonization. Statistical analysis was performed by Fisher's exact test, ANOVA, and Pearson correlation. A P value lower than 0.05 was assumed as statistically significant. Tannerella forsythia was the only periodontal pathogen detected with a statistically admissible frequency. The positivity for Tannerella forsythia was correlated to sampling time and oral hygiene motivation. In buccal epithelial cells, both factors contributed to microbial decrease (P < 0.05), whereas, in crevice, oral hygiene motivation promoted a decrease in the microbial colonization rate (P < 0.05). According to microbiological findings, it is possible to identify how correct motivation for oral hygiene is more than enough to modulate or to avoid an upset of the oral ecosystem balance in early stages of orthodontic treatment.

Oil Essential Mouthwashes Antibacterial Activity against Aggregatibacter actinomycetemcomitans: A Comparison between Antibiofilm and Antiplanktonic Effects.

The aim of this work is to determine the antibacterial activity of three marketed mouthwashes on suspended and sessile states of Aggregatibacter actinomycetemcomitans. The efficacy of two commonly used products in clinical practice, containing essential oils as active ingredients (menthol, thymol, methyl salicylate, and eucalyptol) in association with or without alcohol, has been evaluated in comparison with a chlorhexidine-based mouthwash. The microtiter plate assay, in order to obtain a spectrophotometric measurement of bacterial responses at growing dilutions of each antiseptic, was used for the study. The analysis revealed that a good antibacterial activity is reached when the abovementioned mouthwashes were used at concentration over a 1/24 dilution and after an exposure time of 30 seconds at least. In conclusion, the alcoholic mouthwash appears to have a better biofilm inhibition than its antiplanktonic activity while the nonalcoholic product demonstrates an opposite effect with a better antiplanktonic behavior.

S-100A1 is a reliable marker in distinguishing nephrogenic adenoma from prostatic adenocarcinoma.

Nephrogenic adenoma is a benign lesion that may occur at any site of the genitourinary tract, usually in association with previous urothelial injuries. Although its pathogenesis is still debated, recent studies seem to confirm its derivation from renal tubular epithelium, rather than from a metaplastic process of urothelium. In addition to its uncertain origin, there can be diagnostic difficulty in distinguishing nephrogenic adenoma from prostatic adenocarcinoma, particularly with lesions arising in the prostatic urethra. So far, immunohistochemical stains are often needed to make such a distinction, and several markers have been proposed, often with controversial results. S100A1 is a calcium binding protein that has been recently reported to be expressed in renal tubular cells and in a subset of renal cell neoplasms. Alpha-methylacyl-CoA racemase (AMACR), a recently identified prostate cancer marker, has also been found to be expressed in renal tubules and in some renal epithelial neoplasms. In this study, we investigated the expression of S100A1 and AMACR in 18 nephrogenic adenomas and in 100 prostatic adenocarcinomas. A strong and distinct cytoplasmic or nucleocytoplasmic staining of S100A1 was found in 17 out of 18 cases of nephrogenic adenoma (94%), but never in prostatic adenocarcinoma. In contrast, AMACR expression was detected in 14 of 18 nephrogenic adenomas (78%) and in 96 of 100 prostatic adenocarcinomas (96%). We conclude that (1) S100A1 is a specific and sensitive immunohistochemical marker to differentiate nephrogenic adenoma from prostatic adenocarcinoma; (2) AMACR immunostaining does not seem to be a useful marker in distinguishing between these 2 lesions; (3) given that both S100A1 and AMACR have been reported to be expressed in renal tubular cells and in a subset of renal cell neoplasms, our findings confirm the histogenetic relationship between nephrogenic adenoma and renal tubular epithelium.