Botrytis species affect over 300 host plants in all climate areas of the world, at both pre and post-harvest stages, leading to significant losses in agricultural produce. Therefore, the development of a rapid, sensitive and reliable method to assess the pathogen load of infected crops can help to prescribe an effective curing regime. Growers would then have the ability to predict and manage the full storage potential of their crops and thus provide an effective disease control and reduce post-harvest losses. A highly sensitive electrochemical immunosensor based on a screen-printed gold electrode (SPGE) with onboard carbon counter and silver / silver chloride (Ag/AgCl) pseudo-reference electrode was developed in this work for the detection and quantification of Botrytis species. The sensor utilised a direct sandwich enzyme-linked immunosorbent assay (ELISA) format with a monoclonal antibody against Botrytis immobilised on the gold working electrode. Two immobilisation strategies were investigated for the capture antibody, and these included adsorption and covalent immobilisation after self-assembled monolayer formation with 3-dithiodipropionic acid (DTDPA). A polyclonal antibody conjugated to the electroactive enzyme horseradish peroxidase (HRP) was then applied for signal generation. Electrochemical measurements were conducted using 3,3’, 5,5’-tetramethylbenzidine dihydrochloride / hydrogen peroxide (TMB/H2O2) as the enzyme substrate system at a potential of -200 mV. The developed biosensor was capable of detecting latent Botrytis infections 24 h post inoculation with a linear range from 150 to 0.05 μg fungal mycelium ml-1 and a limit of detection (LOD) as low as 16 ng ml-1 for covalent immobilisation and 58 ng ml-1 for adsorption, respectively. Benchmarked against the commercially available Botrytis ELISA kits, the optimised immuno-electrochemical biosensor showed strong correlation of the quantified samples (R2=0.998).

Orientador: Regina Helena Garcia Martins / Banca: Eulália Sakano / Banca: Vilma Terezinha Anselmo-Lima / Banca: Norimar Hernandez Dias / Banca: Noeme Rocha de Souza / Banca: Regina Helena Garcia Martins / Resumo: O câncer de cabeça e pescoço corresponde a 5% de todas as neoplasias malignas do organismo, sendo os sítios mais prevalentes a cavidade oral, faringe e laringe. O principal fator de risco é o tabagismo e o carcinoma espinocelular é o tipo histológico mais frequente. Marcadores de células tumorais como o p53 e Ki-67 têm sido estudados em tumores de cabeça e pescoço pela sua imunoexpressão positiva para as células neoplásicas e em proliferação. Estudos experimentais possibilitam avaliar os efeitos nocivos do tabaco sobre as vias aéreas, sendo este o objetivo deste estudo. OBJETIVOS: Estudar, em ratos, os efeitos da exposição aguda à fumaça de cigarro sobre as mucosas da língua, faringe e laringe, por meio de análise histológica, morfométrica e imunohistoquímica da expressão das proteínas p53 e Ki-67. MATERIAL E MÉTODO: Ratos Wistar foram subdivididos em dois grupos de 20 animais cada: grupo controle (GC), ração e água ad libitum e grupo tabaco (GT) expostos à inalação de fumaça de 40 cigarro/dia por 60 dias. Os animais foram eutanaziados e realizado biópsias da língua, faringe e laringe para os estudos histopatológico, histomorfométrico e imunohistoquímico das proteínas p53 e do ki-67. RESULTADO: As análises de microscopia de luz da língua dos animais do grupo tabaco (GT) revelaram algumas alterações mais marcantes nesse grupo em cada sítio, sendo que na língua, ganharam destaque a hiperplasia epitelial (GT-90%), hiperplasia de células da camada basal (GT-95%) e displasia epitelial de grau leve a moderado (GT- 85%); na faringe, destacaram-se a hiperplasia de células da camada basal... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: INTRODUCTION: The head and neck cancer represents 5% of all malignant neoplasm of the body, with the sites most prevalent oral cavity, pharynx and larynx. The main risk factor is smoking and squamous cell carcinoma is the most frequent histological type. Tumor cell markers such as p53 and ki-67 have been studied in head and neck tumors by their positive immunostaing for proliferating and neoplastic cells. Experimental studies to assess the possible harmful effects of smoking on the airways which are the objective of this study. OBJECTIVES: To study the effects of acute exposure to cigarette smoke on the mucosa of the tongue, pharynx and larynx in rats; using histological, morphometric and immunohistochemical expression of p53 and ki- 67. MATERIALS AND METHODS: Wistar rats were divided into two groups of 20 animals each control group (CG), food and water ad libitum and tobacco group (GT) exposed to inhalation of smoke from 40 cigarettes/day for 60 days. The animals were euthanized and biopsies of the tongue; pharynx and larynx were performed for histological studies, histomorphometric and immunohistochemical study of p53 and ki- 67. RESULTS: The light of the tongue microscopy analysis showed in GT group: epithelial hyperplasia (GT-90%), hyperplasia of the basal cell layer (GT-95%) and dysplasia of mild to moderate (GT-85%); in pharynx, highlighted the hyperplasia of the basal cell layer (GT-85%), dysplasia (GT-25%) and vascular congestion (GT-95%); predominated in the larynx cell hyperplasia of the basal layer (GT-70%), epithelial hyperplasia (GT-55%), congestion (GT-100%) and inflammatory infiltration of polymorphonuclear neutrophils (GT-25%).The morphometric study identified high measures of the keratin layer in group... (Complete abstract click electronic access below) / Doutor

Os microRNAs (miRNAs, miRs) são pequenos RNAs não codificadores presentes em diferentes organismos. Esses RNAs regulam a tradução de genes alvos por meio de ligação seqüência-específica a RNAs mensageiros (mRNAs). Dependendo do grau de complementaridade, podem inibir a tradução e/ou induzir a degradação desses mRNAs. No presente estudo, foi investigado por PCR em tempo real o padrão de expressão de quatro microRNAs (miR-21, -205, -342 e let-7a ) em quatro linhagens celulares derivadas de tumores da cavidade oral e da faringe (FaDu, Hep-2, SCC9 e UM-SCC-38), em queratinócitos orais normais e em amostras de tumor e margens cirúrgicas pareadas de 34 pacientes com carcinomas epidermóides de cabeça e pescoço (CECP). Foi também investigada a correlação da expressão dos MiRs de interesse com as características clinicopatológicas de pacientes com CECP. Nas linhagens celulares, os níveis dos miRs foram similares ou mais baixos que os de queratinócitos normais, ou os miRs não se expressaram. Somente o miR-342 mostrou níveis elevados na linhagem FaDu. Em células Hep-2 tratadas com estradiol, a expressão de miR-let-7a mostrou-se reduzida. Em tumores primários, níveis baixos de miR-let-7a foram observados em carcinomas de soalho de boca e laringe. A expressão de miR-21, -205 e -342 mostrou grande variabilidade entre as amostras e foi reduzida em um dos sítios anatômicos. Não foi observada correlação entre a expressão dos miRs e as características clinicopatológicas dos pacientes com CECP. A análise de três genes alvo candidatos (LYZ, MGLL e SPRR3) mostrou, em carcinomas de soalho de boca e laringe, associação positiva entre a expressão de miR-let-7a e de seu alvo predito MGLL, uma lipase que pode favorecer o fenótipo maligno aumentando os níveis de ácidos graxos livres e sinais lipídicos oncogênicos. O significado dessa associação não pode ser deduzida dos experimentos realizados pelo presente trabalho. / MicroRNAs (miRNAs, miRs) are small, non-coding RNAs present in different organisms. They regulate the translation of target genes through sequence specific binding to mRNA. Depending on the degree of sequence complimentary, they can inhibit translation and/or degradation of target mRNAs In the present study, we used real time PCR to investigate the expression pattern of four microRNAs (miR-21, -205, -342 e let-7a ) in four cell lines derived from tumors of oral cavity and pharinx (FaDu, Hep-2, SCC9 e UM-SCC-38), in normal oral keratinocytes and in matched tumor / surgical margin samples from 34 patients with head and neck squamous cell carcinomas (HNSCC). We also aimed to correlate the miR expression with the clinicopathological features in HNSCC. In cell lines, the miR levels were similar or lower than those in normal keratinocytes, or even absent. Only miR-342 showed high levels in FaDu cell line. In Hep-2 cells treated with estradiol, miR-let-7a expression was reduced. In primary tumors, low miR-let-7a levels were observed in floor of the mouth and larynx carcinomas. The expression of miR-21, -205 and -342 showed high variability between samples and was reduced in one anatomical site. No correlation was observed between miR expression and clinopathological features of head and neck cancer patients. The analysis of three potential target genes (LYZ, MGLL e SPRR3) showed, in floor of the mouth and larynx carcinomas, a positive correlation between the expression of miR-let-7a and its predicted target gene MGLL, a lipase that may support the malignant phenotype by increasing levels of free fatty acids and oncogenic lipid signals. The meaning of such association was not clear from our data.

Orientadores: Paulo Henrique Ferreira Caria, Darcy de Oliveira Tosello / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-11T04:44:19Z (GMT). No. of bitstreams: 1 Santos_CristinaBellinidos_M.pdf: 1617219 bytes, checksum: 88c2ee3f2eb7ee7363575f7aff1c08cf (MD5) Previous issue date: 2008 / Resumo: Os tipos mais comuns de câncer de cabeça e pescoço são os de pele, boca e laringe, ocorrendo principalmente na população masculina, acima dos cinqüenta anos. O tratamento do câncer de cabeça e pescoço, utilizando a cirurgia, quimioterapia e radioterapia, apresenta seqüelas que afetam funções básicas como a respiração, deglutição, fonação e a mobilidade do segmento cervical da coluna vertebral e ombros. Considerando as alterações sofridas pelo aparelho fonador e na movimentação cervical, o presente estudo realizado em forma de capítulos, teve como objetivos: avaliar o comportamento dos músculos esternocleidomastóideos e paraespinhais cervicais em pacientes com câncer de cabeça e pescoço, submetidos a diferentes ressecções cirúrgicas do tumor; avaliar a eficiência de um protocolo de reabilitação fisioterápico dos músculos cervicais e avaliar o comportamento desses músculos na fala esofágica e com uso da laringe eletrônica durante a fonação. Participaram desse estudo vinte e dois voluntários, idade média de cinqüenta e nove anos, divididos em grupos de acordo com os diferentes objetivos do estudo: avaliação do comportamento dos músculos cervicais de pacientes submetidos a diferentes intervenções cirúrgicas: grupo 1 (laringectomizados totais), grupo 2 (ressecção cirúrgica), grupo 3 (controle); avaliação do protocolo de reabilitação fisioterápica: cinco voluntários com limitações nos movimentos do pescoço, presença de dor e anteriorização da cabeça. O protocolo foi aplicado, duas vezes ao dia, durante um mês, composto de alongamento e relaxamento dos músculos cervicais; avaliação da atividade dos músculos cervicais na fonação: grupo 1: (laringe artificial), grupo 2: (voz esofágica), grupo 3 (controle). Para isso foi utilizado a goniometria e a eletromiografia no repouso, fonação e movimentos cervicais de flexão, extensão, lateralidade direita e esquerda, inclinação direita e esquerda. Resultados: Houve alteração no padrão de ativação muscular nos movimentos de lateralidade direita do músculo esternocleidomastóideo esquerdo e inclinação direita do músculo esternocleidomastóideo direito, independente do tipo de procedimento cirúrgico, com aumento nos valores de RMS (Root Mean Square) relacionados ao grupo controle. Após aplicação do protocolo de tratamento houve aumento da amplitude em todos os movimentos cervicais alterando o padrão de ativação muscular no repouso para os músculos paraespinhais esquerdo, diminuindo os valores de RMS e no movimento de lateralidade direita para o músculo esternocleidomastóideo esquerdo, aumentando os valores de RMS. A fonação não alterou o padrão de ativação muscular dos indivíduos que utilizam a voz esofágica e a laringe artificial, com relação ao grupo controle, no entanto, no repouso houve diferença significativa comparando os valores dos grupos com o grupo controle, para o músculo esternocleidomastóideo direito e músculos paraespinhais direito. Conclusão: Existe alteração no padrão de ativação muscular onde o lado de maior acometimento tumoral e cirúrgico oferece maior resistência para a realização dos movimentos, podendo promover aumento nos valores dos RMS, independente do tipo de cirurgia realizado. O protocolo fisioterápico foi efetivo na recuperação das amplitudes dos movimentos do pescoço podendo alterar o padrão de ativação muscular, principalmente no lado de maior exploração cirúrgica. O uso da laringe artificial ou a fala esofágica não alterou a atividade dos músculos esternocleidomastóideos e paraespinhais cervicais bilateralmente / Abstract: The most common head and neck cancer types are the skin, mouth and larynx cancers, occurring mainly in males, older than fifty years old. The head and neck cancer treatment, employing surgery, chemical therapy, and radiotherapy, presents sequels that affect basic functions as breathing, deglutition, phonation and the mobility of the cervical segment of vertebral column and shoulder. Taking into consideration the changes suffered by the phonatory apparatus and cervical movement, the present study accomplished in form of chapters it had as objectives: to evaluate the behavior of sternocleidomastoid and cervical paraspinal muscles in patients with head and neck cancer submitted to different surgery resections of the tumor; evaluate the efficiency of a physical therapic rehabilitation of cervical muscles protocol and evaluate these muscles behavior in esophageal speech and with the use of electronic larynx during phonation. In this study, twenty two volunteers have participated, in an average age of fifty nine, divided in groups, according to the different objectives of the study: evaluation of the cervical muscles behavior in patients submitted to different surgery interventions: group 1 (total laryngectomy), group 2 (surgery resection), group 3 (control); evaluation of a physical therapic rehabilitation protocol: five volunteers with neck movement limitations, pain and anteriorization of the head. A protocol was applied, twice a day, composed of cervical muscle stretching and relaxing; evaluation of cervical muscles activities in phonation: group 1: (artificial larynx); group 2: (esophageal speech), group 3 (control). For this purpose it has been employed goniometry and the electromyography in repose, in phonation and cervical movements of flexion, extension, left and right laterality, left and right inclination. Results: There was alteration in the muscular activation pattern in the right laterality movements, of left sternocleidomastoid muscle and right inclination of right sternocleidomastoid muscle, regardless the surgery procedure type, with the increase in RMS (Root Mean Square) values related to the control group. After applying the treatment protocol, there was a width increase in all cervical movements changing muscular activation pattern in repose for left paraspinal muscles decreasing values of RMS and in the right laterality movement for left sternocleidomastoid muscle, increasing the RMS values. The phonation, has not changed the muscular activation pattern of individuals that use esophageal speech and the artificial larynx, related to the control group, however, during repose there was a significant difference comparing the groups values to control group, for right sternocleidomastoid muscle and right paraspinal muscles. In conclusion: There is a change in muscular activation pattern where the largest tumor and surgery assault offers a bigger resistance to the movement performance, could promote increase in RMS values, regardless the kind of surgery performed. The physical therapic protocol was effective in recovering the neck movement could alter the muscular activation pattern, mainly in largest surgery assaulted side. The artificial larynx use or the esophageal speech didn't change the activity of sternocleidomastoid muscle and bilaterally cervical paraspinal / Mestrado / Anatomia / Mestre em Biologia Buco-Dental

Alterações no padrão normal de metilação do DNA têm sido caracterizadas como um importante mecanismo na gênese de neoplasias. Esta modificação do DNA é denominada de epigenética uma vez que altera o padrão de expressão das células sem alterar a seqüência do DNA. No câncer, as alterações epigenéticas observadas consistem na hipermetilação das ilhas CpG nos promotores dos genes acompanhada de uma hipometilação global dos dinucleotídeos CpG dispersos pelo genoma. Este evento mostra geralmente ser câncer-específico, ou seja, alguns genes que são metilados em um tipo de câncer, não o são na maioria dos outros tipos. O objetivo deste projeto foi identificar, através da construção de bibliotecas subtrativas de RaSH, genes silenciados por metilação nas linhagens de câncer de cabeça e pescoço FaDu, UM-SCC-14A, UM-SCC-17A e UM-SCC-38 e que possuem expressão induzida após o tratamento com o agente demetilante 5-aza-2-deoxicitidina. Uma vez que a metilação leva a diminuição gradual da expressão gênica, o método RT-PCR semi-quantitativo foi utilizado para validação da expressão diferencial dos genes candidatos PLAU, CD82, RBBP4, AOF2, TMSB10, HSPA5 e LAMC2 nas linhagens não tratadas e tratadas com o agente demetilante 5-aza-2-deoxicitidina. Para todos os genes candidatos foi observado aumento na expressão gênica após o tratamento em pelo menos uma das quatro linhagens. Na linhagem UM-SCC-14 A, os genes CD82, RBBP4, AOF2, HSPA5 e LAMC2 mostraram aumento na expressão após o tratamento com o agente demetilante, sendo que o gene LAMC2 também mostrou esse aumento de expressão na linhagem UM-SCC-17A. Na linhagem UM-SCC-38A todos os genes mostraram aumento de expressão após o tratamento. Embora novos estudos sobre a metilação da região promotora dos genes selecionados sejam necessários, aumentam as evidências de que os genes selecionados sejam regulados pelo mecanismo de metilação e que estejam metilados nas linhagens estudadas. / Abnormalities on the normal pattern of DNA methylation have been caracterized as an important mechanism on carcinogenesis. This modification is called epigenetic and can be defined as a heritable change in gene expression that is not accompanied by changes in DNA sequence. The epigenetic alterations observed on cancer include hypermethylation of selected CpG island gene promoters and simultaneous global hypomethylation. The aim of this project was to identify, by rapid subtraction hybridization, genes silenced by methylation on head and neck cancer lineages with alterations on gene expression after the treatment with 5-aza-2-deoxycytidine. The cancer lineages we used for our experiments were: FaDu, UM-SCC-14A, UM-SCC-17A e UM-SCC-387A and seven genes (PLAU, CD82, RBBP4, AOF2, TMSB10, HSPA5 and LAMC2) were analysed by semiquantitative RT-PCR and for all of them an increaseament of gene expression was observed. For the UM-SCC-14A lineage, the genes CD82, RBBP4, AOF2, HSPA5 and LAMC2 were upregulated after the treatment with demethylating agent as well as LAMC2 was uperegulated on UM-SCC-17A. For the UM-SCC-38A lineage all the genes showed increased expression after the treatmente with -aza-2-deoxycytidine. Our work is another evidence that some genes may be regulated by methylation during carcinogenesis.

Orientador: Wilkens Aurélio Buarque e Silva / Dissertação (mestrado) - Universidade Estadual de Campinas, Faculdade de Odontologia de Piracicaba / Made available in DSpace on 2018-08-19T17:19:18Z (GMT). No. of bitstreams: 1 Fernandes_JoaoPaulodosSantos_M.pdf: 1949531 bytes, checksum: 8b69749dcdca84f0eaba7a2ac05299a1 (MD5) Previous issue date: 2012 / Resumo: O objetivo neste trabalho foi analisar a influência da dimensão vertical de oclusão na postura da coluna cervical e da cabeça por meio de aferições de medidas angulares craniocervicais. Foram selecionados 17 voluntários desdentados totais, com sinais clínicos de diminuição de dimensão vertical de oclusão, portadores de sinais e sintomas de disfunção temporomandibular e usuários de próteses totais, inscritos no cadastro de pacientes do CETASE (Centro de Estudos e Tratamento das Alterações Funcionais do Sistema Estomatognático) da Faculdade de Odontologia de Piracicaba. Os voluntários utilizaram o aparelho de cobertura oclusal plana para o restabelecimento da dimensão vertical de oclusão e do tônus muscular, por um período de 120 dias. Telerradiografias em norma lateral foram realizadas antes (com as próteses totais antigas) e após o período de 120 dias. Foram traçadas as linhas Sela-Násio (SN), a Linha Odontoídea (OD), a Linha CVT (Tangente da Vértebra Cervical), e o Plano mandibular (LM). Cinco medidas angulares craniocervicais foram realizadas: SN/OD; SN/LM; SN/CVT; LM/OD e OD/CVT e duas medidas lineares foram feitas: S-OD e S-CVT. Os dados obtidos foram submetidos à análise estatística pelos testes t de Student e teste de Wilcoxon pareado com nível de significância de p<0,05. As médias iniciais e finais dos ângulos SN/LM e LM/OD apresentaram diferenças estatisticamente significantes, alterando a postura da mandíbula em relação à coluna cervical e à base do crânio e as médias iniciais e finais dos ângulos SN/OD, SN/CVT e OD/CVT não apresentaram diferenças estatisticamente significantes, mostrando o movimento de extensão da cabeça após o tratamento instituído. Os resultados demonstraram que existe relação entre DVO, postura da cabeça e da coluna cervical e que a terapêutica com aparelhos oclusais interferiu melhorando o posicionamento da cabeça e da coluna cervical / Abstract: The aim of this study was to analyze since the influence of the vertical dimension of occlusion in the posture of the cervical spine and head through measurements of craniocervical angles. Seventeen volunteers edentulous were selected, with clinical signs of reduced vertical dimension of occlusion, with signs and symptoms of temporomandibular disorders and denture wearers. The volunteers used occlusal plane appliance coverage for the reestablishment of vertical dimension of occlusion and muscle tone, for a period of 120 days. Lateral cephalometric radiographs were taken before (with the old dentures), and after 120 days. In the radiographs were traced sella-nasion line (SN), Odontoid Line (OD), the CVT line (Cervical Vertebra Tangent), and mandibular plane (ML). Five craniocervical angle measurements were performed: SN/OD, SN/ML, SN/CVT, ML/OD and OD/CVT and two linear measurements were made: S-OD and S-CVT. The data were statistically analyzed by Student's t test and Wilcoxon test with a significance level of p <0.05. The average initial and final angles of the SN/LM and LM/OD showed statistically significant differences by changing the posture of the mandible in relation to the cervical spine and skull base and the mean initial and final angles of the SN/OD, SN/CVT and OD/CVT did not show statistically significant differences, showing the extension movement of the head after the treatment. Our studies demonstrated that a relationship exists between VDO, head posture and cervical spine and that therapy with occlusal devices interfered improving the positioning of the head and cervical spine / Mestrado / Protese Dental / Mestre em Clínica Odontológica

M.Tech. / Purpose: This study aims to compare the effects of chiropractic manipulation versus mobilisation on Pressure Pain Threshold in chronic posterior mechanical cervical spine pain sufferers with regards to pain, disability and cervical spine range of motion. These effects were evaluated using a questionnaire consisting of a McGill Pain Questionnaire, and a Vernon – Mior Neck Pain and Disability Questionnaire, and by measuring cervical spine range of motion using a Goniometer as well as Algometer readings over the restricted facet joint/s in the cervical spine. The questionnaires were completed and the range of motion readings and algometer readings were taken prior to treatment on the first, fourth and seventh consultations. Method: Thirty participants who met the inclusion criteria were divided into two groups of equal size (15 participants each). Group one received spinal manipulation to restricted cervical spine joint/s. The second group received spinal mobilisation to restricted cervical spine joint/s. Participants were treated six times out of a total of seven sessions, over a maximum three week period. Procedure: Subjective data was collected at the beginning of the first and fourth consultations, as well as on the seventh consultation by means of a McGill Pain Questionnaire, and a Vernon – Mior Neck Pain and Disability Questionnaire in order to assess pain and disability levels. Objective data was collected at the beginning of the first and fourth session, as well as on the seventh consultation by means of a Goniometer and Algometer in order to assess cervical spine range of motion and to measure the Pressure Pain Threshold at the restricted facet joint/s in the cervical spine. Analysis of collected data was performed by a statistician. Results: Clinically significant improvements in group 1 and group 2 were noted over the duration of the study with reference to pain, disability, and cervical spine range of motion. Statistically significant changes were noted in group 1 and group 2 with reference to pain and disability, and in group 1 and group 2 with v reference to certain cervical spine range of motions as well as algometer readings to measure the Pressure Pain Threshold at the restricted facet joint/s. Conclusion: The results show that both spinal manipulation and mobilization are effective treatment protocols (as demonstrated clinically, and to a lesser extent, statistically) in decreasing pain and disability, and increasing cervical spine range of motion and most importantly Pressure Pain Threshold at the restricted facet joint/s in patients with chronic posterior mechanical cervical spine pain. Although the study did not allow for a definite conclusion to be drawn, the results suggest that Chiropractic manipulative therapy is an effective treatment protocol to increase the Pressure Pain Threshold in chronic posterior mechanical neck pain sufferers. The advantage of this is that the treatment modality is used to its full potential, thereby providing the patient with the best results in terms of lasting benefits. It also shows that in cases where manipulation is contra-indicated mobilization will have a similar effect, but the long term benefits are questionable.

M.Tech. / Purpose: This study was undertaken in order to demonstrate the effects of static magnetic field therapy on acupuncture points in the treatment of those suffering from mechanical neck pain. Isolated spinal manipulative therapy of the cervical spine was compared to spinal manipulative therapy of the cervical spine in conjunction with magnetic field therapy on acupuncture points using both objective and subjective measurements. Before the execution of this study, it was hypothesised that both treatment protocols would be effective in the treatment of mechanical neck pain, although the combined therapy would be more effective. Method: Patients were recruited by way of advertisements placed in and around the University of Johannesburg, Doornfontein campus and their health clinic. Thirty patients with mechanical neck pain were recruited and randomly divided into two groups. Group one received manipulation to the affected joints of the cervical spine and group two received manipulation to the cervical spine combined with magnetic field therapy on acupuncture points. Procedure: Each patient in each group attended six treatment sessions; three in the first week and three in the second week. The Vernon Mior Neck Pain and Disability Index and the Numerical Pain Rating Scale (subjective measurements) were completed by each patient and the Cervical Range of Motion instrument (objective measurements) was used to collect readings from each patient in both the control and experimental groups, subjective and objective measurements were taken before treatment one, three and six. Specific treatment protocols were then adhered to. Results: The results indicated that both treatment protocols were effective in reducing mechanical neck pain although not one group was more effective than VI the other. Both groups improved subjectively and objectively as they had cervical spinal manipulation directed at joint dysfunction. Conclusion: The experimental group who received spinal manipulative therapy to correct joint dysfunction in conjunction with magnetic field therapy on acupuncture points was not more effective than the control group who received spinal manipulation only, in the treatment of mechanical neck pain.

Abstract Traditional clinicopathological factors are not accurate enough to predict the behavior of head and neck squamous cell carcinoma (HNSCC). The most powerful indicator of prognosis is the stage of the disease. New prognostic markers have, however, been searched for in order to better identify patient groups in need of different treatments or follow-up. Gelatinases (MMP-2, -9) are endopeptidases associated with tumor invasion and angiogenesis, and their tissue inhibitors (TIMP-1, -2) are also linked to cancer cell invasion and metastasis formation. In some cancer types they are even prognostic and relate with a more aggressive clinical course of the disease. In the present work the expression and the clinical significance of tumor tissue and circulating immunoreactive proteins for MMP-2, -9, TIMP-1 and -2 were assessed in HNSCC. The study group included 74 patients with HNSCC and 44 healthy controls. The expression of immunoreactive proteins was examined in paraffin-embedded tumor sections by immunohistochemical staining using specific antibodies, and the pretreatment serum levels of those proteins were quantitatively measured by ELISA assay. Immunohistochemical overexpression of MMP-9 in tumor was for the first time found to predict the prognosis for shortened survival in HNSCC, the cause-specific survival rates being 45% and 92% and relapse-free survival being 42% and 79% in MMP-9 positive or negative cases, respectively. Additionally, tissue TIMP-1, MMP-2 and TIMP-2 positivity were all also linked with poorer survival of patients with HNSCC. However, these differences remained less distinct than with MMP-9. The expression of gelatinases and their inhibitors in tumor tissue was also an indicator for later lymph node or hematogenic relapses in HNSCC patients. Circulating MMP-9 and TIMP-1 levels were significantly higher in HNSCC patients than in healthy controls. Further, the cause-specific and relapse-free survival rates were lower among HNSCC patients with high MMP-9 and TIMP-1 serum levels compared to patients with low levels of circulating MMP-9 and TIMP-1. A significant correlation was shown between circulating MMP-9 and MMP-9 immunohistochemical staining in the corresponding tumors. No correlation was found between tissue or circulating levels of gelatinases or their inhibitors and the traditional clinical or histopathological factors, except for the association between tissue and circulating TIMP-1 and the size of the primary tumor. Taken together, these results suggest that tissue expression of gelatinases and their inhibitors as well as pretreatment circulating MMP-9 and TIMP-1 levels could be prognostic in estimation of the clinical course of HNSCC. The results indicate further studies are needed with larger patient materials.

Botrytis species affect over 300 host plants in all climate areas of the world, at both pre and post-harvest stages, leading to significant losses in agricultural produce. Therefore, the development of a rapid, sensitive and reliable method to assess the pathogen load of infected crops can help to prescribe an effective curing regime. Growers would then have the ability to predict and manage the full storage potential of their crops and thus provide an effective disease control and reduce post-harvest losses. A highly sensitive electrochemical immunosensor based on a screen-printed gold electrode (SPGE) with onboard carbon counter and silver / silver chloride (Ag/AgCl) pseudo-reference electrode was developed in this work for the detection and quantification of Botrytis species. The sensor utilised a direct sandwich enzyme-linked immunosorbent assay (ELISA) format with a monoclonal antibody against Botrytis immobilised on the gold working electrode. Two immobilisation strategies were investigated for the capture antibody, and these included adsorption and covalent immobilisation after self-assembled monolayer formation with 3-dithiodipropionic acid (DTDPA). A polyclonal antibody conjugated to the electroactive enzyme horseradish peroxidase (HRP) was then applied for signal generation. Electrochemical measurements were conducted using 3,3’, 5,5’-tetramethylbenzidine dihydrochloride / hydrogen peroxide (TMB/H2O2) as the enzyme substrate system at a potential of -200 mV. The developed biosensor was capable of detecting latent Botrytis infections 24 h post inoculation with a linear range from 150 to 0.05 μg fungal mycelium ml-1 and a limit of detection (LOD) as low as 16 ng ml-1 for covalent immobilisation and 58 ng ml-1 for adsorption, respectively. Benchmarked against the commercially available Botrytis ELISA kits, the optimised immuno-electrochemical biosensor showed strong correlation of the quantified samples (R2=0.998) ... [cont.].