Clinical and pathological features affecting cardiac sympathetic denervation in autopsy-confirmed dementia with Lewy bodies.

BACKGROUND AND PURPOSE: The aim was to clarify the features affecting cardiac sympathetic denervation in autopsy-confirmed dementia with Lewy bodies (DLB) patients. METHODS: Fifty-four autopsy-confirmed DLB patients were enrolled. Tissue samples of the left ventricular anterior wall were immunostained with anti-tyrosine hydroxylase antibody to identify catecholaminergic nerve axons. Immunostained areas were quantified as residual cardiac sympathetic nerve (CSN) axons and the relationship between the degree of residual CSN axons and clinical and neuropathological features was examined. RESULTS: Virtually all patients showed small amounts of residual CSN axons (0.87%, range 0.02%-9.98%), with 50 patients (92.6%) showing <2.0% of residual axons. The patients who showed psychological symptoms within the first year of the disease had significantly more residual CSN axons than the remaining patients did (1.50% vs. 0.40%, P < 0.01). Patients with a short disease duration and neocortical-type Lewy body pathology tended to have more preserved CSN axons, although this difference was not statistically significant. Fifty-three patients (98.1%) who had neurofibrillary tangles in the brain and strong concomitant Alzheimer's disease pathology also had statistically significantly more preserved CSN axons. The patient with the most preserved CSN axons showed different characteristics from the results, except for the first symptom. CONCLUSION: Psychological symptoms within the first year of the disease, a short disease duration, neocortical-type Lewy body pathology and strong concomitant Alzheimer's disease pathology may be related to mild CSN degeneration in DLB patients. Thus, DLB patients with broad Lewy body pathology in the brain in the early stages may show mild CSN degeneration.

Serum retinol levels are associated with brain volume loss in patients with multiple sclerosis.

BACKGROUND: Although predicting future brain volume loss (BVL) in patients with multiple sclerosis (MS) is important, studies have shown only a few biomarkers that can predict BVL. OBJECTIVES: The aim of this study is to elucidate the association between longitudinal BVL and serum biomarker candidates. METHODS: This single-center, retrospective, observational study intended to cover MS patients during January 2008 to March 2016. Patients who underwent brain MRI two times at intervals of >24 months and had a blood test to measure biomarker candidates at the time or within three months of the MRI scan were included. Evaluation of brain volume was performed by using SIENAX and SIENA in the FMRIB software library. RESULTS: Twenty-three patients with MS were included in this study. We found that serum retinol binding protein (RBP) levels were significantly correlated with percentage brain volume change (PBVC) (p = 0.0079). Furthermore, best subset selection of multiple linear regression models identified baseline normalized brain volume and serum RBP as the best predictors of PBVC. CONCLUSIONS: Our study shows that lower serum retinol levels are associated with greater longitudinal BVL and that serum RBP and can be a predictor of BVL.

Microfluidic immunomagnetic cell separation using integrated permanent micromagnets.

In this paper, we demonstrate the possibility to trap and sort labeled cells under flow conditions using a microfluidic device with an integrated flat micro-patterned hard magnetic film. The proposed technique is illustrated using a cell suspension containing a mixture of Jurkat cells and HEK (Human Embryonic Kidney) 293 cells. Prior to sorting experiments, the Jurkat cells were specifically labeled with immunomagnetic nanoparticles, while the HEK 293 cells were unlabeled. Droplet-based experiments demonstrated that the Jurkat cells were attracted to regions of maximum stray field flux density while the HEK 293 cells settled in random positions. When the mixture was passed through a polydimethylsiloxane (PDMS) microfluidic channel containing integrated micromagnets, the labeled Jurkat cells were selectively trapped under fluid flow, while the HEK cells were eluted towards the device outlet. Increasing the flow rate produced a second eluate much enriched in Jurkat cells, as revealed by flow cytometry. The separation efficiency of this biocompatible, compact micro-fluidic separation chamber was compared with that obtained using two commercial magnetic cell separation kits.

Early changes in carcinogenesis of colorectal adenomas.

OBJECTIVES: Adenocarcinoma of the colon and rectum is the third most common cause of cancer deaths and the sixth most common cancer in the world. Adenomas are benign neoplastic lesions which can be transformed into carcinomas, but this is usually not the case. There should be some risk factors which lead to the development of carcinomas into adenomas. The aim of this study is to find out the early changes and high risk factors related to carcinogenesis in colonic polyps. METHODS: In this study, we reviewed nearly 1000 colonoscopic biopsies and chose 72 biopsies. We developed three groups (tubular adenomas group 1, villous adenomas group 2, normal mucosa group 3); each group had 24 different biopsies. P53, Ki-67, bcl-2, cyclin D1, E-cadherin, c-erb B2 immunohistochemistry and human papillomavirus (HPV) in-situ hybridization were used for analysis. RESULTS: Five of the seventy-two cases were positive in HPV in-situ analysis. Four of them were villous adenomas and one was a tubular adenoma. Ki-67 expression was limited only to crypts in group 3 but in groups 1 and 2, Ki-67 expression was seen both in crypt epithelium and surface epithelium. Cyclin D1, c-erb B2, bcl-2 expression was significantly increased in neoplastic polyps. CONCLUSION: Ki-67 expression, both in the crypt and surface epithelium, and cyclin D1, c-erb B2, bcl-2 over-expression may be a clue of dysplastic epithelium and if the role of HPV is elucidated and shown to be important in colonic carcinogenesis, then vaccination might prevent carcinogenesis caused by HPV.

Early changes in carcinogenesis of colorectal adenomas / Cambios tempranos en la carcinogénesis de los adenomas colorectales

OBJECTIVES: Adenocarcinoma of the colon and rectum is the third most common cause of cancer deaths and the sixth most common cancer in the world. Adenomas are benign neoplastic lesions which can be transformed into carcinomas, but this is usually not the case. There should be some risk factors which lead to the development of carcinomas into adenomas. The aim of this study is to find out the early changes and high risk factors related to carcinogenesis in colonic polyps. METHODS: IIn this study, we reviewed nearly 1000 colonoscopic biopsies and chose 72 biopsies. We developed three groups (tubular adenomas group 1, villous adenomas group 2, normal mucosa group 3); each group had 24 different biopsies. P53, Ki-67, bcl-2, cyclin D1, E-cadherin, c-erb B2 immunohistochemistry and human papillomavirus (HPV) in-situ hybridization were used for analysis. RESULTS: Five of the seventy-two cases were positive in HPV in-situ analysis. Four of them were villous adenomas and one was a tubular adenoma. Ki-67 expression was limited only to crypts in group 3 but in groups 1 and 2, Ki-67 expression was seen both in crypt epithelium and surface epithelium. Cyclin D1, c-erb B2, bcl-2 expression was significantly increased in neoplastic polyps. CONCLUSION: Ki-67 expression, both in the crypt and surface epithelium, and cyclin D1, c-erb B2, bcl-2 over-expression may be a clue of dysplastic epithelium and if the role of HPV is elucidated and shown to be important in colonic carcinogenesis, then vaccination might prevent carcinogenesis caused by HPV.

OBJETIVOS: El adenocarcinoma del colon y recto es la tercera causa más común de muertes por cáncer y el sexto tipo de cáncer más común en el mundo. Los adenomas son lesiones neoplásicas benignas que pueden transformarse en carcinomas, pero éste normalmente no es el caso. Debe haber algunos factores de riesgo que conducen al desarrollo de carcinomas en adenomas. El objetivo de este estudio es averiguar los cambios tempranos y los factores de alto riesgo relacionados con la carcinogénesis en los pólipos colónicos. MÉTODOS: En este estudio, revisamos casi 1000 biopsias colonoscópicas y escogimos 72 biopsias. Desarrollamos tres grupos (grupo 1: adenomas tubulares, grupo 2: adenomas vilosos, grupo 3: mucosa normal); cada grupo tuvo 24 biopsias diferentes. Para el anílisis se utilizaron la inmunohistoquímica de P53, Ki-67, bcl-2, ciclina D1, E-cadherina, y c-erb B2, así como la hibridación in situ para la detección del virus del papiloma humano (VPH) RESULTADOS: Cinco de setenta y dos casos resultaron positivos en el análisis del VPH in-situ. Cuatro de ellos fueron adenomas vilosos, de los cuales uno era un adenoma tubular. La expresión Ki-67 está limitada sólo a las criptas en el grupo 3, pero en los grupos 1 y 2, la expresión Ki-67 se observó tanto en el epitelio de la cripta como en el epitelio de la superficie. La expresión de la ciclina D1, c-erb B2, y bcl- 2 se halla significativamente aumentada en los pólipos neoplásicos. CONCLUSIÓN: La expresión de Ki-67 tanto en el epitelio de la cripta como de la superficie, y la sobre-expesión de la ciclina D1, c-erb B2, bcl-2 puede ser una clave para el epitelio displásico, y si se aclara y demuestra que el papel del VPH es importante en la carcinogénesis colónica, entonces la vacunación podría prevenir los carcinogénesis inducidos por el VPH.

Tunable and label-free bacteria alignment using standing surface acoustic waves.

This paper describes a new technique for focusing bacteria in a microfluidic channel and subsequently controlling their trajectory. Bacteria alignment is obtained using standing surface acoustic waves (SSAW) generated by two interdigitated transducer electrodes (IDTs) patterned on a piezoelectric wafer. The bacteria are focused in the standing wave pressure nodes, separated by half a wavelength, the electrode geometry and applied voltage frequency being chosen accordingly. Interestingly, the position of a pressure node can be modulated by introducing a phase shift between the electrical signals applied to both IDTs. The bacteria, trapped in this node, follow it and can therefore be deflected. This technique works with label-free bacteria in their culture medium and induces low power consumption, which is very interesting for portable devices.

Diagnostic pitfalls in the evaluation of fine needle aspiration cytology of the thyroid: correlation with histopathology in 260 cases.

OBJECTIVES: Fine needle aspiration cytology (FNAC) of the thyroid is a non-invasive, cost-effective screening procedure that is valuable for distinguishing neoplastic lesions from non-neoplastic nodules. The aim of this study was to determine the diagnostic accuracy of FNACs performed at our institution by correlating FNAC results with histopathological diagnoses. METHODS: Two hundred and seventy-one aspiration cytology specimens followed by thyroidectomy were included in the study, and the results of 260 adequate FNACs were compared with their histological diagnoses. RESULTS: The sensitivity and specificity of thyroid FNAC for detecting neoplasia were 92.6% and 91.6%, respectively. There were 15 (5.7%) false positives and six (2.3%) false negatives. CONCLUSIONS: The results showed that follicular cells that exhibit some of the features of papillary carcinoma could be observed in a cytology slide of Hashimoto's thyroiditis, leading to a diagnostic pitfall. In addition, cellularity and overlapping cytological criteria in hyperplasia might lead to a false diagnosis.

A clinical, genetic, and neuropathologic study in a family with 16q-linked ADCA type III.

Presented is the new kindred with autosomal dominant cerebellar ataxia linked to chromosome 16q22.1 (16q-ADCA type III) associated with progressive hearing loss. By haplotype analysis, the critical interval was slightly narrowed to three megabase regions between GATA01 and D16S3095. Neuropathologic study of 16q-ADCA type III demonstrated characteristic shrinkage of Purkinje cell bodies surrounded by synaptophysin-immunoreactive amorphous material containing calbindin- and ubiquitin-positive granules.

Somatosensory-evoked cortical potential during attacks of paroxysmal dysesthesia in multiple sclerosis.

Paroxysmal dysesthesia is considered to be one of the characteristic symptoms of multiple sclerosis (MS), but the lesion responsible and the pathophysiology of this dysesthesia are not known. We report the interesting finding of somatosensory-evoked potentials (SEPs) in a patient with MS during a paroxysmal dysesthesia attack.

Cytoplasmic and nuclear polyglutamine aggregates in SCA6 Purkinje cells.

Aggregations of the alpha1A-calcium channel protein have been previously demonstrated in spinocerebellar ataxia type 6 (SCA6). Here the authors show that small aggregates, labeled by a monoclonal antibody 1C2 that preferentially detects expanded polyglutamine larger than that in SCA6 mutation, are present mainly in the cytoplasm but also in the nucleus of Purkinje cells. Although the length of expansion is small in SCA6, the current finding might indicate that SCA6 conforms to the pathogenic mechanism(s) in other polyglutamine diseases.

Morphologic evaluation and integrin expression profile of renal tubular cells cultured from percutaneous renal biopsy specimen.

Kidney biopsy is an indispensible procedure for making a pathologic diagnosis of renal diseases by fixing and staining the biopsy specimen. However, it is not a routine procedure to culture the cells from a renal biopsy specimen directly, or to utilize the cultured cells for any kind of diagnostic or functional evaluation. In this study, primary culture of the renal tubular epithelial cells was tried from a piece of percutaneous kidney biopsy specimen. Successive passages of the cells were possible until fourth passage. With these cells, morphologic characteristics of the cultured cells and integrin expression profiles were investigated. On light and electron microscopy, these cells were characterized by the cobblestone-like growth, presence of microvilli and tight junction, and the preservation of polarity. Immunohistochemical studies demonstrated the epithelial nature of these cells and particularly their differentiation from renal tubular epithelial cells, of either proximal or distal nephronic segment. The integrin profile confirms the epithelial nature of the cell. We hope that our results facilitate the understanding of pathophysiology of renal tubular cells from the patient directly.

Ataxin-3 is translocated into the nucleus for the formation of intranuclear inclusions in normal and Machado-Joseph disease brains.

Machado-Joseph disease (MJD)/spinocerebellar ataxia type 3 (SCA3) is one of the dominantly inherited cerebellar ataxias. The gene responsible for the disease, a novel gene of unknown function, encodes ataxin-3 containing a polyglutamine stretch. Although it has been known that ataxin-3 is incorporated into neuronal intranuclear inclusions (NIIs) in neurons of affected regions, the relationship between NII formation and neuronal degeneration still remains uncertain. In the present study we show two different conditions in which ataxin-3 is recruited into the nucleus and suggest a process to form nuclear inclusions. In normal brains, wild-type ataxin-3 localizes within the ubiquitin-positive nuclear inclusion, the Marinesco body, indicating that ataxin-3 is recruited into the nuclear inclusion even in the absence of pathologically expanded polyglutamine. In MJD/SCA3 brains, immunohistochemical analyses with anti-ataxin-3 antibody, anti-ubiquitin antibody, and monoclonal antibody 1C2 known to recognize expanded polyglutamine revealed differences in frequency and in diameter among NIIs recognized by each antibody. These results were confirmed in the same inclusions by double immunofluorescent staining, suggesting that expanded ataxin-3 forms a core, thereby recruiting wild-type ataxin-3 into the nucleus around the core portion, and then followed by activation of the ubiquitin/ATP-dependent pathway. Recruitment of ataxin-3 into the nucleus and formation of nuclear inclusion under two different conditions suggest that ataxin-3 may be translocated into the nucleus under certain conditions stressful on neuronal cells such as aging and polyglutamine neurotoxicity.

Spinocerebellar ataxia type 6 mutation alters P-type calcium channel function.

Abnormal CAG repeat expansion in the alpha1A voltage-dependent calcium channel gene is associated with spinocerebellar ataxia type 6, an autosomal dominant cerebellar ataxia with a predominant loss of the Purkinje cell. A reverse transcriptase-polymerase chain reaction analysis of mRNA from mouse Purkinje cells revealed a predominant expression of the alpha1A channel lacking an asparagine-proline (NP) stretch in the domain IV (alpha1A(-NP)). Human alpha1A channels carrying various polyglutamine length with or without NP were expressed in HEK293 cells, and channel properties were compared using a whole-cell voltage clamp technique. alpha1A(-NP), corresponding to P-type channel, with 24 and 28 polyglutamines found in patients showed the voltage dependence of inactivation shifting negatively by 6 and 11 mV, respectively, from the 13 polyglutamine control. Contrarily, the alpha1A channel with NP (alpha1A(+NP)), corresponding to Q-type channel, with 28 polyglutamines exhibited a positive shift of 5 mV. These results suggest that altered function of alpha1A(-NP) may contribute to degeneration of Purkinje cells, which express predominantly alpha1A(-NP), due to the reduced Ca(2+) influx resulting from the negative shift of voltage-dependent inactivation. On the other hand, other types of neurons, expressing both alpha1A(-NP) and alpha1A(+NP), may survive because the positive shift of voltage-dependent inactivation of alpha1A(+NP) compensates Ca(2+) influx.

Abundant expression and cytoplasmic aggregations of [alpha]1A voltage-dependent calcium channel protein associated with neurodegeneration in spinocerebellar ataxia type 6.

Spinocerebellar ataxia type 6 (SCA6) is one of the eight neurodegenerative diseases caused by a tri-nucleotide (CAG) repeat expansion coding polyglutamine (CAG repeat/polyglutamine diseases) and is characterized by late onset autosomal dominant cerebellar ataxia and predominant loss of cerebellar Purkinje cells. Although the causative, small and stable CAG repeat expansion for this disease has been identified in the [alpha]1A voltage-dependent calcium channel gene (CACNA1A), the mechanism which leads to predominant Purkinje cell degeneration is totally unknown. In this study, we show that the calcium channel mRNA/protein containing the CAG repeat/polyglutamine tract is most intensely expressed in Purkinje cells of human brains. In SCA6 brains, numerous oval or rod-shaped aggregates were seen exclusively in the cytoplasm of Purkinje cells. These cytoplasmic inclusions were not ubiquitinated, which contrasts with the neuronal intra-nuclear inclusions of other CAG repeat/polyglutamine diseases. In cultured cells, formation of perinuclear aggregates of the channel protein and apoptotic cell death were seen when transfected with full-length CACNA1A coding an expanded polyglutamine tract. The present study indicates that the mechanism of neurodegeneration in SCA6 is associated with cytoplasmic aggregations of the [alpha]1A calcium channel protein caused by a small CAG repeat/polyglutamine expansion in CACNA1A.