Holocarboxylase synthetase deficiency induced by HLCS gene mutations: a rare disease study / 中国当代儿科杂志

A boy, aged 16 months, attended the hospital due to head and facial erythema for 15 months and vulva erythema for 10 months with aggravation for 5 days. The boy developed perioral and periocular erythema in the neonatal period and had erythema and papules with desquamation and erosion in the neck, armpit, and trigone of vulva in infancy. Blood gas analysis showed metabolic acidosis; the analysis of amino acid and acylcarnitine profiles for inherited metabolic diseases and the analysis of organic acid in urine suggested multiple carboxylase deficiency; genetic testing showed a homozygous mutation of c.1522C>T(p.R508W) in the HLCS gene. Finally the boy was diagnosed with holocarboxylase synthetase deficiency and achieved a good clinical outcome after oral biotin treatment. This article analyzes the clinical data of a child with holocarboxylase synthetase deficiency and summarizes the etiology, diagnosis, and treatment of this child, so as to provide ideas for clinicians to diagnose this rare disease.

Expert consensus on screening, diagnosis and treatment of multiple carboxylase deficiency / 浙江大学学报·医学版

Multiple carboxylase deficiency (MCD) includes autosomal recessive holocarboxylase synthetase (HLCS) deficiency and biotinidase (BTD) deficiency, which are caused by and gene mutations respectively. Neonatal screening for HLCS deficiency is based on 3-hydroxyisovaleryl carnitine in dry blood filter paper, and BTD deficiency is based on BTD activity determination. HLCS deficiency and BTD deficiency are characterized by neurocutaneous syndrome and organic aciduria, however, they are different in onset age, neurological symptoms and metabolic decompensation, which needed to be differentiated from acquired biotin deficiency or other genetic metabolic diseases. The diagnosis of the disease requires a combination of biochemical characteristics of hematuria, enzyme activity determination and genetic test. Routine biotin doses are effective for most MCD patients. This consensus is intended to benefit early screening and diagnosis of MCD.

Hipoventilação relacionada ao sono de origem central secundária à deficiência de biotinidase: relato de caso / Sleep-related hypoventilation due to partial biotinidase deficiency: a case repor

RESUMO: A hipoventilação relacionada ao sono de origem central resulta em hipercapnia relacionada ao sono na vigência de condições normais do sistema respiratório e excluindo-se outros fatores. Os pacientes portadores dessa patologia podem se apresentar assintomáticos ou com queixas de cefaleia matinal, déficit cognitivo e fadiga, além de eventos como a observação de respiração superficial. No presente relato, descreve-se o caso de uma paciente de três anos, com exame físico geral e neurológico normais, desenvolvimento neuropsicomotor adequado, apresentando irregu-laridades respiratórias e bradicardia durante o sono. Encaminhada para investigação de distúrbios respiratórios do sono, sendo diagnosticada com hipoventilação relacionada ao sono. Através do estudo genético, evidenciou-se a deficiência de biotinidase como a possível causa da sintomatologia, comprovada por dosagens enzimáticas e teste genético molecular. O tratamento medicamentoso foi iniciado precocemente, determinando resolução dos sintomas descritos. A importância do presente relato se encontra na apresentação da deficiência da biotinidase com quadro cardiorrespiratório isolado em criança neurologicamente normal, ademais trata-se de um caso em que a etiologia de Breath-Holding Spells foi a deficiência dessa enzima. Correspondência sugerida pela resolução da hipoventila-ção central após a introdução da biotina. Além disso, nesse caso, os sintomas Apparent Life-Threatening Events, que aterrorizam o observador e até o profissional, foram solucionados com tratamento simples, a ingesta oral de biotina. Esse relato de caso corrobora com a expansão das possibilidades de manifestações fenotípicas das formas tardias de deficiência de biotinidase, como o fenótipo da Síndrome da Hipoventilação Central. (AU)

ABSTRACT: Idiopathic sleep-related hypoventilation occurs in individuals with hypercapnia during sleep in normal conditions of the respiratory system in the absence of other disorders. Patients with this condition may be asymptomatic or have complaints of morning headache, cognitive deficit and fatigue, and observation of shallow breathing. This report describes the case of a 3-year-old patient with normal physical and neurological exam, appropriate neuropsychomotor development, presenting breathing irregularities, and bradycardia during sleep. The patient was referred to an investigation for sleep respiratory disturbs and was diagnosed with hypoventilation related to sleep. The genetic study, done by enzymatic dosages and molecular genetic tests, showed the deficiency of biotinidase as a possible cause of symptomatology. The drug treatment was initiated early with the resolution of the symptoms. The present clinical report highlights the biotinidase deficiency with an isolated cardiorespiratory condition in a neurologically normal child, which also led to Breath-Holding Spells. This relation was suggested after central hypoventilation resolution following biotin introduction. Besides, Apparent Life-Threatening Events symptoms, which terrify the observer and even professionals, disappeared after the oral intake of biotin. Finally, this case report corroborates the expansion of possibilities for the phenotypic manifestations of late cases from biotinidase deficiency, as the SHC phenotyp. ((AU)

Comparison of Biotin Interference in Second- and Third-Generation Roche Free Thyroxine Immunoassays

No abstract available.

Long Non-Coding RNA LINC00525 Promotes the Stemness and Chemoresistance of Colorectal Cancer by Targeting miR-507/ELK3 Axis

BACKGROUND AND OBJECTIVES: This study aims to explore the effects of a long non-coding RNA, LINC00525, on colorectal cancer (CRC) and its underlying molecular mechanisms. METHODS: The qPCR, MTT, colony formation, Western blotting, Luciferase reporter and biotin pull-down, shRNA knockdown and DNA fragmentation assays were performed in this study. RESULTS: High expressions of LINC00525 were associated with poor prognosis of CRC patients. LINC00525 knockdown decreased stemness properties and increased sensitivities to oxaliplatin. MiR-507 was a direct target of LINC00525 and overexpression of miR-507 significantly decreased abilities of tumorsphere formation and cell growth. Overexpression of miR-507 resulted in a decrease of expression of cancer stem cell markers and the increase of apoptosis rates. MiR-507 regulated the expression of ELK3. In addition, LINC00525 knockdown decreased the expression of ELK3. Restoration of ELK3 expression abrogated the effects of LINC00525 knockdown. LINC00525 could be served as prognostic marker of CRC. CONCLUSIONS: LINC00525 enhanced stemness properties and increased sensitivities of CRC cells to oxaliplatin by targeting miR-507/ELK3 axis.

Comparative study of biotin supplementation on weight gain and occurrence of digital diseases in cattle (Bos taurus x Bos indicus) / Estudo comparativo da suplementação com biotina sobre o ganho de peso e ocorrência de enfermidades digitais em bovinos (Bos taurus x Bos indicus)

This study aimed to assess the weight gain and the incidence of foot diseases in male, crossbred (Bos taurus x Bos indicus) bovines that were supplemented with biotin. An amount of 240 animals, supplemented or not with biotin, allocated in 12 groups of 20 animals, was assessed for a period of six months. The study was conducted during three years and the groups were divided according to the forage available, corn, corn residue and sorghum silage, and initial weights between 100 and 200 kg and between 200 and 300 kg. The statistical analyses used were the Tukey's Test, in triple factorial scheme (type of silage x use of biotin x initial body weight) and the Fisher´s Exact Test, both at 5% significance level. The biotin supplementation in bovines did not influence weight gain and the incidence of foot diseases, however, when comparing only the type of forage, corn and sorghum silage provided higher weight gain than silage made of corn residue.

Esse estudo teve como objetivo avaliar o ganho em peso e a ocorrência de enfermidades digitais em bovinos do sexo masculino, mestiços (Bos taurus x Bos indicus) que foram suplementados com biotina. Avaliaram-se, por um período de seis meses, 240 bovinos, suplementados ou não com biotina, alocados em 12 grupos de 20 animais. O estudo foi realizado durante três anos e os grupos foram divididos de acordo com o volumoso disponibilizado, silagem de milho, de resíduo de milho e de sorgo, e pesos iniciais entre 100 e 200 kg e entre 200 e 300 kg. As análises estatísticas empregadas foram o Teste de Tukey, em esquema de fatorial triplo (tipo de silagem x uso da biotina x peso corporal inicial) e o Teste Exato de Fisher, ambas em nível de significância de 5%. A suplementação com biotina nos bovinos não exerceu influência sobre o ganho em peso e a ocorrência de enfermidades digitais, mas, quando se comparou apenas o tipo de volumoso, a silagem de milho e sorgo, pode se observar um maior ganho em peso que a silagem confeccionada de resíduo de milho.

Differential Inhibition of Nav1.7 and Neuropathic Pain by Hybridoma-Produced and Recombinant Monoclonal Antibodies that Target Nav1.7 : Differential activities of Nav1.7-targeting monoclonal antibodies / 神经科学通报·英文版

The voltage-gated Na channel subtype Nav1.7 is important for pain and itch in rodents and humans. We previously showed that a Nav1.7-targeting monoclonal antibody (SVmab) reduces Na currents and pain and itch responses in mice. Here, we investigated whether recombinant SVmab (rSVmab) binds to and blocks Nav1.7 similar to SVmab. ELISA tests revealed that SVmab was capable of binding to Nav1.7-expressing HEK293 cells, mouse DRG neurons, human nerve tissue, and the voltage-sensor domain II of Nav1.7. In contrast, rSVmab showed no or weak binding to Nav1.7 in these tests. Patch-clamp recordings showed that SVmab, but not rSVmab, markedly inhibited Na currents in Nav1.7-expressing HEK293 cells. Notably, electrical field stimulation increased the blocking activity of SVmab and rSVmab in Nav1.7-expressing HEK293 cells. SVmab was more effective than rSVmab in inhibiting paclitaxel-induced mechanical allodynia. SVmab also bound to human DRG neurons and inhibited their Na currents. Finally, potential reasons for the differential efficacy of SVmab and rSVmab and future directions are discussed.

Development of Cookies with Brewer's Yeast and Beans to Improve Skin Health of Lactating Women

The purpose of this research was to provide basic information for cookies made with black soybeans, chick peas, lentils, oatmeal, and brewer's yeast and to establish the optimum formula for the development of low glycemic index (GI) cookies with high biotin content for lactating women. This study was performed to determine the optimal composite recipe of oatmeal cookies with two different concentrations levels of bean powder (black soybeans, chick peas, lentils) and brewer's yeast using a central composite design. In addition, the mixing conditions of oatmeal cookies were optimized using response surface methodology of sensory evaluation and mechanical and physicochemical analysis. As a result, mechanical and physicochemical analyses showed significant values for lightness, redness, yellowness, hardness, and water content (P < 0.05), while sensory evaluation showed significant values for flavor, taste, crispness, and overall acceptability (P < 0.05). The optimal sensory combination was suggested to be 3.73 g of bean powder and 1.59 g of brewer's yeast. Considering all outcomes obtained throughout the experiments, brewer's yeast, black soybeans, chick peas, lentils, and oatmeal are suitable ingredients for increasing functionality and consumer acceptability of cookies. In addition, these results are expected to be useful in producing cookies of optimal quality, contributing to the development of various nutritious foods, and improving the food industry for lactating women.

Development of 9(th) Revision Korean Food Composition Table and Its Major Changes / 대한지역사회영양학회지

OBJECTIVES: The Korean Food Composition Table (KFCT) was first published in 1970, and has since been updated every five years by the Rural Development Administration (RDA). This study was conducted to introduce the development strategies, features, and challenges of the 9th revision of the KFCT. METHODS: Due to the increasing demands of nutrient database users and generators, the RDA started a new research project in 2013 to improve the quantity and quality of data for the 9th revision of the KFCT. Over 1,000 food items frequently consumed in Korea were selected as key foods using the results of the Korean National Health and Nutrition Examination Survey. About 200 raw materials and processed food items were collected and analyzed every year. Target nutrients that were analyzed by collaborative labs, such as, sugars, selenium, iodine, and biotin, were increased from 22 to 43. Analytical sample handling procedures and data quality evaluation systems were also established in collaboration with 10 contracted labs. Data were evaluated for data quality according to the FAO/INFOODS, CODEX, and AOAC guidelines. RESULTS: The 9th revision contains data on 3,000 food items and up to 43 and 140 food nutrients for the printed table and the excel database file, respectively. Overall, 1,485 data items were newly added, 973 of which were provided by the RDA and 512 were cited from foreign nutrient databases. The remaining 1,515 food items were maintained as in the 8th revision. CONCLUSIONS: The KFCT provides the basic infrastructure for food and nutrition policy, research, and dietary practice in South Korea. The use of the KFCT has increased exponentially in the past few years in both public and private sectors; accordingly, increased efforts should be paid to the preparation, improvement, and maintenance of KFCT.

Development of 9(th) Revision Korean Food Composition Table and Its Major Changes / 대한지역사회영양학회지

OBJECTIVES: The Korean Food Composition Table (KFCT) was first published in 1970, and has since been updated every five years by the Rural Development Administration (RDA). This study was conducted to introduce the development strategies, features, and challenges of the 9th revision of the KFCT. METHODS: Due to the increasing demands of nutrient database users and generators, the RDA started a new research project in 2013 to improve the quantity and quality of data for the 9th revision of the KFCT. Over 1,000 food items frequently consumed in Korea were selected as key foods using the results of the Korean National Health and Nutrition Examination Survey. About 200 raw materials and processed food items were collected and analyzed every year. Target nutrients that were analyzed by collaborative labs, such as, sugars, selenium, iodine, and biotin, were increased from 22 to 43. Analytical sample handling procedures and data quality evaluation systems were also established in collaboration with 10 contracted labs. Data were evaluated for data quality according to the FAO/INFOODS, CODEX, and AOAC guidelines. RESULTS: The 9th revision contains data on 3,000 food items and up to 43 and 140 food nutrients for the printed table and the excel database file, respectively. Overall, 1,485 data items were newly added, 973 of which were provided by the RDA and 512 were cited from foreign nutrient databases. The remaining 1,515 food items were maintained as in the 8th revision. CONCLUSIONS: The KFCT provides the basic infrastructure for food and nutrition policy, research, and dietary practice in South Korea. The use of the KFCT has increased exponentially in the past few years in both public and private sectors; accordingly, increased efforts should be paid to the preparation, improvement, and maintenance of KFCT.

Association of Nutrient Intakes with Cognitive Function in Koreans Aged 50 years and Older

This study attempted to investigate whether nutrient and food intake were related with mild cognitive impairment (MCI) in adults and elderly over 50 years of age in Korea. Questionnaires and anthropometric measurements were conducted on general aspects of the research, and food frequency questionnaires (FFQs) were conducted to determine nutritional status. The relative theta power (RTP) through electroencephalography (EEG) measurements, neurocognitive function test (NFT; CNS Vital Signs), and cognitive function was measured. The MCI group consumed significantly lower C18:4, eicosapentaenoic acid (EPA), docosahexaenoic acid (DHA) among the N-3 fatty acids, N-6 fatty acids dihomo-γ-linolenic acid (DGLA), mono unsaturated fatty acids, C22:1, biotin, vitamin D in the nutrients, and sweet potato (12.35g/day, p = 0.015), mackerel (3.38g/day, p = 0.017), mandarin orange (p = 0.016), persimmon (p = 0.013) and apple (p = 0.023) in the food than the normal group did. And the MCI group consumed salted fish (3.14g/day, p = 0.041) and ice-cream (5.01g/day, p = 0.050) at a significantly higher level. Delayed verbal score, delayed visual score, and verbal memory score of the NFT and RTP values of the prefrontal cortex among the EEGs were significantly lower in the MCI group compared to those in the normal group. From this study, we found that nutrient and food intake are closely related to MCI in Korean aged 50 years and older, but more human studies are needed to verify these findings.

Clinical Manifestations, Gene Analysis of Patients with 3-Methylcrotonyl-CoA Carboxylase Deficiency

OBJECTIVE: 3-Methylcrotonyl CoA carboxylase deficiency (3MCCD) is classified as organic acid disease due to leucine catabolism. It is among the most common inborn errors of metabolism identified on newborn screening test using tandem mass spectrometry. There is a broad spectrum of clinical presentations. 3-Methylcrotonyl CoA carboxylase converts 3-methylcrotonyl-CoA to 3-methylglutaconyl-CoA using biotin as a coenzyme in mitochondria. Restricting protein diets and supplementing carnitine, glycine, and biotin are known treatments. We reported this study to find out clinical symptoms, type of gene mutation, and effect of treatment. METHODS: This study was based on retrospective data of patients with 3MCCD in Soonchunhyang University Seoul Hospital and Soonchunhyang University Bucheon Hospital between April 2009 to August 2016. RESULTS: All 10 infants were born term infants and had no symptoms. During the neonatal period, abnormalities were detected in the new born screening test using tandem mass spectrometry, 3-hydroxyisovalerylcarnitine was increased. 3-Methylcrotonylglycine (3MCG) and 3-hydroxyisovalreric acid (3HIVA) were examined in urine organic acid assay. The results showed that 3MCG was increased in all 10 children. Except for three of the 10 children, 3HIVA was increased. Genetic tests were performed on all 10 children. MCCC1 gene mutations were detected in four patients and MCCC2 mutations were detected in six patients. After diagnosis, all children were recommended leucine-restricted diets, and seven of the 10 patients started to feed on leucine free formula for the treatment of 3MCCD. CONCLUSION: According to our data, all patients has no symptoms and are shown normal development. There were no clinical symptoms or changes in prognosis according to gene mutation type.

Relato de dois casos: falso positivo de TRAb causados por uso de biotina para tratamento dermatológico / Two case report: TRAb false positive caused by the use of biotin for dermatological treatment

RESUMO A Biotina é um componente da dieta utilizada em vários suplementos vitamínicos e fórmulas para algumas patologias específicas. Nos últimos anos temse visto através de vários estudos que a biotina tomada em doses altas, em uma suplementação supra fisiológica, tem relação com alterações significativas nos testes de função tiroidiana como T3, T4 e TSH e no TRAb, pela interação da biotina com a estreptavidina em imunoensaios competitivos, e imunométricos ou tipo "sanduíche", levando a uma combinação laboratorial compatível com o hipertireoidismo. Neste trabalho relatamos os casos de duas pacientes que tomaram biotina em dose baixa de 10 mg por três meses para tratamento dermatológico aonde foram vistos aumentos significativas do TRAb, gerando um falso positivo e podendo ser fator de confusão na hora de definir a etiologia do quadro de tireotoxicose por Doença de Graves. A queda de TRAB foi 48 h após a última dose de biotina. A comunidade médica deve ser alertada sobre essa interação e orientar os pacientes sobre a retirada da biotina 10 mg pelo menos 48 hs antes de coletar exames que incluam o TRAb. Palavras-chave: Biotina; Hipertireoidismo; Tireotoxicose; Doença de Graves;

Preparation, characterization, and antiproliferative activities of biotin-decorated docetaxel-loaded bovine serum albumin nanoparticles

ABSTRACT The aim of the present study was to characterize biotin-decorated docetaxel-loaded bovine serum albumin nanoparticles (DTX-BIO-BSA-NPs) and evaluate their antiproliferative activity in vitro. The particle size of prepared DTX-BIO-BSA-NPs was found to be always lower than 200 nm, with sizes of 166.9, 160.3, 159.0, 176.1 and 184.8 nm and the zeta potential was -29.51, -28.54, -36.54, -36.08 and -27.56 mV after redissolution with water for 0, 1, 2, 4 and 8 hours, respectively. The polydispersity index (PDI) was stable in the range of 0.170 - 0.178. In the in vitro drug-release study, the DTX-BIO-BSA-NPs targeted a human breast cancer cell line MCF-7 effectively. The x-ray diffraction spectrum and DSC curve of DTX-BIO-BSA-NPs suggested that docetaxel was in an amorphous or disordered crystalline phase in DTX-BIO-BSA-NPs. In vitro cytotoxicity results showed that DTX-BIO-BSA-NPs inhibits proliferation of MCF-7, SGC7901, LS-174T and A549 cells in a concentration-dependent manner after exposure to DTX-BIO-BSA-NPs for 48 hours. Taken together, these results indicate that DTX-BIO-BSA-NPs may have potential as an alternative delivery system for parenteral administration of docetaxel.

Evaluation of the Menus of Senior Welfare Centers for Home-bound Elderly

This study examined the characteristics of the foodservice menu items offered at senior welfare centers to provide information on Korean senior menu development. A total of 514 lunch menu items were collected from 27 senior welfare centers in April, July, October and January. The most frequently served staple foods, soups, and side dishes were multi-grain rice, seaweed soup, Bulgogi, Kimchi, and liquid yogurt. The proportions of carbohydrate, protein, and lipids of total energy serving of senior welfare centers were 59.8%: 16.7%: and 22.8%, respectively. The nutrients served at less than 40% of the Recommended Nutrient Intake (RNI) and Adequate Intake (AI) of Dietary Reference Intakes for Koreans (KDRIs) were chloride (1.0%), vitamin D (1.3%), biotin (1.7%), magnessium (4.5%), Iodine (7.5%), pantothenic acid (8.0%), vitamin E (12.5%), vitamin B6 (20.0~21.4%), vitamin K (21.1~24.3%), and water (35.7~39.7%). The nutrients served in excess of the daily intake goal and RNI were iron (98.9~127.1%), sodium (104.9%), and copper (1,100.0%).

Evaluación de reactividad inmunohistoquímica con método del Complejo Avidina­Biotina (ABC) / Evaluation of immunohistochemical reactivity with Avidin-Biotin Complex (ABC) method

Inmunohistoquímica es toda técnica que permite detectar in situ componentes celulares y extracelulares por medio de anticuerpos específicos, empleando sistemas de detección enzimáticos. Dentro de los métodos inmunohistoquímicos, la técnica del complejo avidina­biotina(ABC) es ampliamente utilizada debido a su alta sensibilidad. El objetivo del presente estudio fueevaluar la reactividad inmunohistoquímica del anticuerpo 4C4.9 para la detección de la proteínaS-100, utilizando el método ABC. Para la evaluación de la reactividad inmunohistoquímica se utilizaron 2 biopsias de piel humana con diagnóstico histopatológico de melanoma maligno nodular ulcerado y nevus melanocítico intradérmico, provenientes del Laboratorio de Investigación en Biotecnología Animal de la Universidad de La Frontera, Temuco, Chile. Se utilizó el Kit VECTASTAIN®como método de detección, la dilución del anticuerpo 4C4.9 fue 1/250 y la temperatura de incubación fue a 4 ºC ó 37 ºC por 18 horas. Para validar la técnica, se realizó un control positivo y otro negativo para 4C4.9. Los resultados de la tinción inmunohistoquímica por el método del complejo ABC mostraron tinción positiva para la proteína S-100, tanto en melanoma maligno nodular ulcerado, como en nevus melanocítico intradérmico, incubados durante 18 horas a 4 ºC ó 37 ºC. Sin embargo, la inmunotinción fue más intensa cuando el anticuerpo primario se incubó a 37 ºC. Para una correcta interpretación de los resultados, es necesario tener en consideración que la reacción antígeno-anticuerpo se ve influenciada por diversos factores, como la concentración del anticuerpo, el tiempo y la temperatura de incubación. En conclusión, nuestros resultados sugieren incubarlas muestras con el primer anticuerpo (4C4.9) en una dilución de 1/250 en agua destilada, incu-bando durante 18 h a 37 ºC. Se recomienda la utilización del anticuerpo 4C4.9 como apoyo al diagnóstico y diagnóstico diferencial.

Immunohistochemistry is anytechnique that can detect cellular and extracellular components in situ by means of specific antibodies,using enzymatic detection systems. Among immunohistochemical methods, the technique ofavidin - biotin complex (ABC) is widely used because of its high sensitivity. The aim of this study was to evaluate the immunohistochemical reactivity of the4C4.9 antibody for detection of S-100 protein using the ABC method. For the evaluation ofimmunohistochemical reactivity 2 biopsies of humanskin were used with histopathological diagnosis ofulcerated malignant melanoma and melanocyticintradermal nevi from the Research Laboratory onAnimal Biotechnology of the Universidad de La Fron-tera, Chile. The Kit VECTASTAIN® was used asdetection method, the dilution the 4C4.9 antibodywas 1/250 and incubation temperature was at 4 °Cor 37 °C for 18 hours. To validate the technique, apositive control and a negative for 4C4.9 was performed. The results of immunohistochemicalstaining by the method of ABC complex showed positive staining for protein S-100 both in ulcerated malignant melanoma and melanocytic intradermalnevi, incubated for 18 hours at 4 °C or 37 °C.However, immunostaining was more intense when the primary antibody was incubated at 37° C. For acorrect interpretation of the results, it is necessary to take into consideration that the antigen-antibody reaction is influenced by various factors such as the concentration of antibody, time and temperature ofincubation. In conclusion, our results suggest incubating the samples with the first antibody (4C4.9)at 1/250 dilution in distilled water, incubating for 18h at 37 ºC. However, immunostaining was moreintense when the primary antibody was incubated at37° C. For a correct interpretation of the results, it isnecessary to take into consideration that antigen-antibody reaction is influenced by various factors suchas the concentration of antibody, time and temperature of incubation. In conclusion, our results suggest incubating the samples with the first antibody(4C4.9) at 1/250 dilution in distilled water, incubating for 18 h at 37 ºC. The use of the antibody 4C4.9 is recommended to support the diagnosis and differential diagnosis.

Morphometric analysis of feedforward pathways from the primary somatosensory area (S1) of rats

We used biotinylated dextran amine (BDA) to anterogradely label individual axons projecting from primary somatosensory cortex (S1) to four different cortical areas in rats. A major goal was to determine whether axon terminals in these target areas shared morphometric similarities based on the shape of individual terminal arbors and the density of two bouton types: en passant (Bp) and terminaux (Bt). Evidence from tridimensional reconstructions of isolated axon terminal fragments (n=111) did support a degree of morphological heterogeneity establishing two broad groups of axon terminals. Morphological parameters associated with the complexity of terminal arbors and the proportion of beaded Bp vs stalked Bt were found to differ significantly in these two groups following a discriminant function statistical analysis across axon fragments. Interestingly, both groups occurred in all four target areas, possibly consistent with a commonality of presynaptic processing of tactile information. These findings lay the ground for additional work aiming to investigate synaptic function at the single bouton level and see how this might be associated with emerging properties in postsynaptic targets.

Surface interactions between two of the main periodontal pathogens: Porphyromonas gingivalis and Tannerella forsythia

PURPOSE: Porphyromonas gingivalis and Tannerella forsythia have been implicated as the major etiologic agents of periodontal disease. These two bacteria are frequently isolated together from the periodontal lesion, and it has been suggested that their interaction may increase each one's virulence potential. The purpose of this study was to identify proteins on the surface of these organisms that are involved in interbacterial binding. METHODS: Biotin labeling of surface proteins of P. gingivalis and T. forsythia and liquid chromatography-tandem mass spectrometry (LC-MS/MS) analysis was performed to identify surface proteins involved in the coaggregating activity between P. gingivalis and T. forsythia. RESULTS: It was found that three major T. forsythia proteins sized 161, 100, and 62 kDa were involved in binding to P. gingivalis, and P. gingivalis proteins sized 35, 32, and 26 kDa were involved in binding to T. forsythia cells. CONCLUSIONS: LC-MS/MS analysis identified one T. forsythia surface protein (TonB-linked outer membrane protein) involved in interbacterial binding to P. gingivalis. However, the nature of other T. forsythia and P. gingivalis surface proteins identified by biotin labeling could not be determined. Further analysis of these proteins will help elucidate the molecular mechanisms that mediate coaggregation between P. gingivalis and T. forsythia.

Promotion of Pink1S Auto-phosphorylation with CK2β / 生物医学工程学杂志

The aim of this study is to determine the regulatory mechanism of PTEN-induced putative kinase protein 1 short isoform (PINK1S) in cytoplasm. By co-immunoprecipitation (Co-IP) assay, we identified that PINK1S interacted with the beta regulatory subunit of Casein Kinase 2 (CK2β), but not with the catalytic subunits CK2α1 and CK2α2. Furthermore, cells were transfected with PINK1S and CK2β, and then PINK1S was purified by immunoprecipitation. After detecting the phosphorylated proteins by Phos-tag Biotin, we found that CK2β overexpression increased auto-phosphorylation of PINK1S. Finally, we generated CK2β knockdown cell lines by RNA interference. Purified PINK1S from CK2β knockdown cells significantly reduced its auto-phosphorylation compared with control cells. These results suggested that CK2β functions as a regulatory subunit of PINK1S kinase complex promoted its activation by self-phosphorylation.