BPI-fold (BPIF) containing/plunc protein expression in human fetal major and minor salivary glands

Abstract The aim of this study was to determine expression, not previously described, of PLUNC (palate, lung, and nasal epithelium clone) (BPI-fold containing) proteins in major and minor salivary glands from very early fetal tissue to the end of the second trimester and thus gain further insight into the function of these proteins. Early fetal heads, and major and minor salivary glands were collected retrospectively and glands were classified according to morphodifferentiation stage. Expression of BPI-fold containing proteins was localized through immunohistochemistry. BPIFA2, the major BPI-fold containing protein in adult salivary glands, was detected only in the laryngeal pharynx; the lack of staining in salivary glands suggested salivary expression is either very late in development or is only in adult tissues. Early expression of BPIFA1 was seen in the trachea and nasal cavity with salivary gland expression only seen in late morphodifferentiation stages. BPIFB1 was seen in early neural tissue and at later stages in submandibular and sublingual glands. BPIFA1 is significantly expressed in early fetal oral tissue but BPIFB1 has extremely limited expression and the major salivary BPIF protein (BPIFA2) is not produced in fetal development. Further studies, with more sensitive techniques, will confirm the expression pattern and enable a better understanding of embryonic BPIF protein function.

MUC-1 expression in pleomorphic adenomas using two human milk fat globule protein membrane antibodies (HMFG-1 and HMFG-2)

Pleomorphic adenoma (PA) is the most common salivary gland tumor and its microscopic features and histogenesis are a matter of debate. Human milk fat globule protein membrane (HMFG) monoclonal antibodies (MoAbs) comprise a set of antibodies against the mucin 1 (MUC-1) protein detected in several salivary gland tumors. Objective The aim of this study was to assess the immunoexpression of the PA neoplastic cells to MUC-1 protein using HMFG-1 and HMFG-2 MoAbs, contrasting these results with those from normal salivary gland tissue. Material and Methods Immunohistochemical detection of MUC-1 protein using HMFG-1 and HMFG-2 MoAbs was made in 5 mm thick, paraffin embedded slides, and the avidin-biotin method was used. Results Positivity to HMFG-1 and HMFG-2 MoAbs was found in ductal, squamous metaplastic and neoplastic myoepithelial cells, keratin pearls and intraductal mucous material. Two kinds of myoepithelial cells were identified: classic myoepithelial cells around ducts were negative to both MoAbs, and modified myoepithelial cells were positive to both MoAbs. This last cellular group of the analyzed tumors showed similar MUC-1 immunoexpression to ductal epithelial cells using both HMFG antibodies. Intraductal mucous secretion was also HMFG-1 and HMFG-2 positive. Conclusions Our results showed there are two kinds of myoepithelial cells in PA. The first cellular group is represented by the different kinds of neoplastic myoepithelial cells and is HMFG-positive. The second one is HMFG-negative and represented by the neoplastic myoepithelial cells located around the ducts. .

Localization of vesicular inhibitory amino acid transporter (VIAAT) in the submandibular salivary gland throughout the postnatal development of mice / Localización del transportador vesicular de aminoácidos inhibidores (VIAAT) en la glándula salival submandibular durante el desarrollo postnatal de ratones

According to recent studies, it is highly possible that the occurrence of vesicular inhibitory amino acid transporter (VIAAT) is a good marker of GABA-signaling not only in the brain, but also in extra-brain tissue cells containing GABA and GAD. In view of this, the present study was attempted to localize VIAAT-immunoreactivity in the submandibular gland of mice. In the present study, the submandibular glands of male mice at various postnatal developmental stages were examined for detailed localization of VIAAT-immunoreactivity in immunohistochemistry at light microscopic level. The immunoreactivity for VIAAT was localized in epithelial cells of proximal and distal excretory ducts with the striated portion more intensely immunopositive at young postnatal stages. No significant immunoreactivity was seen in the acinar cells throughout the postnatal development. In addition, the immunoreactivity for VIAAT was detected in the salivary parasympathetic ganglionic neurons, but not in any nerve fibers surrounding the glandular cells. Furthermore, VIAAT-immunoreactivity was found in smooth muscle cells forming the outermost layer of intralobular arterioles. From the present findings, it is possible that GABA plays roles as paracrine and autocrine regulators in the saliva secretion as well as the gland development.

Según estudios recientes, es altamente posible que la aparición del transportador vesicular de aminoácidos inhibidores (VIAAT) sea un buen marcador de señalización de GABA no sólo en el cerebro, sino también en células de tejido extra-cerebrales que contienen GABA y GAD. En el presente estudio se intentó localizar inmunoreactividad a VIAAT en la glándula submandibular de ratones. En el presente estudio, se examinaron las glándulas submandibulares de ratones machos en las distintas etapas del desarrollo postnatal para la localización detallada de inmunoreactividad a VIAAT inmunohistoquímicamente a nivel de microscopía óptica. La inmunorreactividad para VIAAT se localizó en las células epiteliales de los conductos excretores proximal y distal, con mayor intensidad en la porción estriada en las etapas tempranas. No se observó inmunoreactividad significativa en las células acinares durante el desarrollo postnatal. Además, se detectó la inmunoreactividad para VIAAT en las neuronas ganglionares parasimpáticas salivales, pero no en las fibras nerviosas que rodean las células glandulares. Además, la inmunoreactividad a VIAAT se encuentra en las células del músculo liso que forman la capa más externa de las arterias interlobulillares. En base a estos hallazgos, es posible que GABA tenga una función como regulador autocrino y paraparacrino en la secreción de saliva, así como en el desarrollo de la glándula.

5-Bromo-2’-deoxyuridine induces visible morphological alteration in the DNA puffs of the anterior salivary gland region of Bradysia hygida (Diptera, Sciaridae)

5-Bromo-2’-deoxyuridine (BrdUrd) has long been known to interfere with cell differentiation. We found that treatment ofBradysia hygida larvae with BrdUrd during DNA puff anlage formation in the polytene chromosomes of the salivary gland S1 region noticeably affects anlage morphology. However, it does not affect subsequent metamorphosis to the adult stage. The chromatin of the chromosomal sites that would normally form DNA puffs remains very compact and DNA puff expansion does not occur with administration of 4 to 8 mM BrdUrd. Injection of BrdUrd at different ages provoked a gradient of compaction of the DNA puff chromatin, leading to the formation of very small to almost normal puffs. By immunodetection, we show that the analogue is preferentially incorporated into the DNA puff anlages. When BrdUrd is injected in a mixture with thymidine, it is not incorporated into the DNA, and normal DNA puffs form. Therefore, incorporation of this analogue into the amplified DNA seems to be the cause of this extreme compaction. Autoradiographic experiments and silver grains counting showed that this treatment decreases the efficiency of RNA synthesis at DNA puff anlages.

Anticoagulant activity in salivary gland homogenates of Thyrsopelma guianense (Diptera: Simuliidae), the primary vector of onchocerciasis in the Brazilian Amazon

In this study, anticoagulant activity was detected in salivary gland homogenates (SGHs) of Thyrsopelma guianense (Diptera: Simuliidae). The SGH yielded 1.07 ìg ± 0.03 (n = 15) of total soluble protein per pair of glands. In addition, following SDS-PAGE (12.5 percent gel) and silver nitrate staining, 12 polypeptides with molecular weights ranging from 14-69 kDa were detected in all physiological ages analyzed (12 h, 24 h, 48 h and 72 h following emergence). Coagulation bioassays showed that the SGHs had activities that interacted at all levels of coagulation (the intrinsic, extrinsic and common pathways), by extending the plasma recalcification time, prothrombin time, thrombin time. This is the first report on the activity of salivary gland proteins from the main vector of onchocerciasis in Brazil. We also suggest detailed studies on the morphology and function of the salivary glands in order to understand the role of these proteins in host/vector interactions.

Salivary gland proteins of the human malaria vector, Anopheles dirus B (Diptera: Culicidae)

Salivary gland proteins of the human malaria vector, Anopheles dirus B were determined and analyzed. The amount of salivary gland proteins in mosquitoes aged between 3 - 10 days was approximately 1.08 ± 0.04 æg/female and 0.1 ± 0.05 æg/male. The salivary glands of both sexes displayed the same morphological organization as that of other anopheline mosquitoes. In females, apyrase accumulated in the distal regions, whereas alpha-glucosidase was found in the proximal region of the lateral lobes. This differential distribution of the analyzed enzymes reflects specialization of different regions for sugar and blood feeding. SDS-PAGE analysis revealed that at least seven major proteins were found in the female salivary glands, of which each morphological region contained different major proteins. Similar electrophoretic protein profiles were detected comparing unfed and blood-fed mosquitoes, suggesting that there is no specific protein induced by blood. Two-dimensional polyacrylamide gel analysis showed the most abundant salivary gland protein, with a molecular mass of approximately 35 kilodaltons and an isoelectric point of approximately 4.0. These results provide basic information that would lead to further study on the role of salivary proteins of An. dirus B in disease transmission and hematophagy.

Proteínas das glândulas salivares do Anopheles dirus B (Diptera: Culicidae), vetor da malária humana foram determinadas e analisadas. A quantidade de proteínas das glândulas salivares em mosquitos com três a 10 dias de idade foi de aproximadamente 1,08 ± 0,04 æg/ fêmea e de 0,1 ± 0,05 æg/macho. As glândulas salivares de ambos os sexos mostraram organização morfológica semelhante à de outros mosquitos anofelinos. Em fêmeas, apirase acumula-se nas regiões distais, enquanto alfa-glucosidase foi encontrada na região proximal dos lóbulos laterais. Esta distribuição diferencial das enzimas analisadas reflete a especialização de diferentes regiões para alimentação de açucares e sangue. Análise SDS-PAGE revelou que pelo menos sete proteínas foram encontradas nas glândulas salivares de fêmeas, das quais cada região morfológica continha diferentes proteínas principais. Perfis eletroforéticos de proteínas semelhantes foram detectados comparando-se mosquitos não alimentados e alimentados por sangue, sugerindo que não existe proteína específica induzida pelo mesmo. Análise por gel poliacrilamida bi-dimensional mostrou a mais abundante proteína de glândulas salivares com aproximadamente 35 kilodaltons de massa molecular e ponto isoelétrico de aproximadamente 4,0. Estes resultados dão informações básicas que levariam a estudos adicionais sobre o papel das proteínas salivares do An. dirus B na transmissão da doença e hematofagia.

study on the effects of different temperatures on protein changes in the salivary glands of hyalomma anatolicum anatolicum and boophilus annulatus

It was determined whether ambient temperature influences the proteins produced by salivary glands of Hyalomma anatolicum analolicum and Boophilus annulatus. Unfed adults of H. anatolicum anatolicum and partly fed female adult of B. annulatus were subjected to different temperatures [4, 15, 28 and 42 C] and proteins in soluble denatured salivary gland extracts [SGE] analysed by SDS-PAGE. The authors noticed changes during different temperatures but 29, 55, and 97 KDa protein bands remained unchanged at all temperatures of two species and in B. annulatus 29, 36, 55, 66, 84, 97 and 205 KDa proteins were constant

Biochemical fingerprints of salviary gland for some ticks species in Egypt by SDS-polyacrylamide gel electrophoresis

SDS-PAGE gel electrophoresis as a sensitive method is used for simultaneously comparing the total protein extracted from salivary glands in some ticks species [Argas persicus, Hyalomma antolicum excavatum, H. an antolicum, H. dromedarii, H. impeltatum and Rhipicephalus sanguineus]. The results of A. persicus were characterized by 4 bands with molecular weights ranging from 82.76 to 16.076 KD. On the other hand, H. an. excavatum females were characterized by 6 bands with molecular weights ranging from 103.74 to 6.216 KD. H. an. antolicum males were characterized by 5 bands with molecular weights ranging from 79.183 to 2.092 KD. R. sanguineus was characterized by 4 and 8 bands with molecular weights ranging from 88.864 to 6.216 KD and 126.69 to 7.1 12 KD for male and female, respectively. Female H. impeltatum was characterized by 8 bands with a range of molecular weights from 106.96 to 7.829 KD. H. dromedarii was characterized by 4 and 7 bands with ranges of molecular weights from 88.147 to 4.602 KD and 110.20 to 2.630 KD for male and female, respectively

Clinical and biochemical evaluation of the saliva of patients with xerostomia induced by radiotherapy

Foram avaliados alguns aspectos clínicos e algumas propriedades bioquímicas salivares de 21 pacientes, antes e após o tratamento radioterápico para câncer de cabeça e pescoço (grupo experimental) e de 21 pacientes sem câncer (grupo controle). O fluxo salivar foi avaliado pelo tempo necessário (segundos) para produção estimulada de 2 ml de saliva e a capacidade tamponante determinada frente à utilização de um método colorimétrico simples. A concentração de proteína total salivar foi determinada pelo método de Bradford4. A atividade da amilase foi mensurada através dos açúcares redutores liberados e quantificados pelo método do ácido dinitrossalicílico utilizando a glicose como substrato. O perfil eletroforético foi avaliado em gel de poliacrilamida (SDS-PAGE 12%) para amostras salivares contendo 5 mg de proteína. Foi observada, no grupo experimental, redução estatisticamente significativa (p < 0,01) para o fluxo salivar (162,47 s ± 28,30 antes e 568,71 s ± 79,75 após) e para a capacidade tamponante (pH 5,45 ± 0,14 antes e 4,40 ± 0,15 após). Não foi observada alteração estatisticamente significativa na concentração de proteína. A atividade específica da a-amilase foi significativamente diminuída (p < 0,01) (856,6 ng/mg ± 88,0 antes e 567,0 ng/mg ± 120,6 após). No perfil eletroforético, foram observadas diferenças nas bandas protéicas, principalmente na faixa de peso molecular de 72.000 a 55.000 Da. Clinicamente, os pacientes com xerostomia induzida pela radioterapia apresentaram aumento de lesões na mucosa.

Dissociation between vasodilation and Leishmania infection-enhancing effects of sand fly saliva and maxadilan

In this study, the ability of maxadilan and Lutzomyia longipalpis salivary gland lysate to enhance the infection of CBA mice by Leishmania major and of BALB/c mice by L. braziliensis was tested. No difference was observed between sizes of lesion in CBA mice infected with L. major and treated or not with salivary gland lysate or maxadilan, although they were injected in concentrations that induced cutaneous vasodilation. Although parasites were more frequently observed in foot pads and spleens of animals treated with maxadilan than in the animals treated with salivary gland lysate or saline, the differences were small and not statistically significant. The lesions in BALB/c mice infected with L. braziliensis and treated with maxadilan were slightly larger than in animals that received Leishmania alone. Such differences disappeared 14 weeks after infection, and were statistically significant only in one of two experiments

Avaliação sialométrica e sialoquímica de indivíduos submetidos à radioterapia na região de cabeça e pescoço / Sialometric and sialochimistry evaluation of indivíduos submithed to the x-ray in the region of head and neck

Os danos provocados pela radiações ionizantes ao funcionamento das glândulas salivares são reconhecidos háquase 90 anos. Os mecanismos responsáveis pela destruição do parêquina da glândula ainda permanecem incertos, no entanto, a saliva manifesta alterações de natureza qualitativa e quantitativa que refletem na qualidade de vida do indivíduo irradiado. A proposta deste estudo foi verificar as alterações icorridas no fluxodo pH e na composição orgânica e ionorgânica da saliva humana durante e após um protocolo padronizado de radioterapia na região da cabeça e pescoço. Amostras de saliva foi totalmente estimulada de um grupo de indivíduos foram, então, coletadas ante, durante e por até 6 meses depois do tratamento radioterápico. O fluxo, pH e as concentrações de fosfato, calcio, sódio, magnésio, potássio, ferro, tiocianato, cloretos, amilase, proteínas totais, creatinina, uréia, ácido úrico, lactato, colesterol e glicose da saliva foram analisadas. Esta mesma avaliação sialométrica e sialoquímica foi realizada num grupo de indivíduos saudáveis e noutro representado por pacientes portadores de neoplasia maligna na região da cabeça e do pescoço ainda não submetido a qualquer intervenção terapeutica. Os efeitos da radiação sobre a saliva já puderam ser demonstrados após os indivíduos receberaem a dose de 15 Gy. O fluxo salivar sofreu uma redução significativa de 50 por cento durante o tratamento (15 Gy) e diminuiu em cerca de 79 porcento aos seis meses do pós-tratamento (p<0,05). O pH da saliva também baixou de 7,6 para 6,8 ao final do tratamento radioterápico (p<0,05). Já as concentrações de calcio, sódio, potássio, cloretos e magnésio elevaram-se significativamente, enquanto que a concentração de fosfato diminui (p<0,05). Os componentes orgãnicos da saliva não manifestaram mudanças significativas. Toda esta série de modificações ocorridas na parte orgânica e aquosa da saliva dos indivíduos irradiados demonstram que esta secreção, embora, mais concentrads do que a saliva de indivíduos saudáveis, não desempenham as suas funções normais. Sendo assim, todos os indivíuos submetidos a radioterapia na região da cabeça e pescoço por campos paralelos, opostos e laterais, cujas glândulas salivares estejam incluídas na área tratada, requerem um planejamento sistemático de cuidados odontológicos que devem ter início antes da radioterapia, mantido durante o tratamento e continuado para o resto de sua vida.

Prenatal development of human major salivary glands. Histological and immunohistochemical characteristics with reference to adult and neoplastic salivary glands

No abstract available.

Aspectos citoquímicos dos mucocitos de glândulas linguais de bufo ictericus, spix 1824 (amphibia, anura) / Cytochemical aspectos of the mucocytes of lingual glands of bufo ictericus, spix 1824 (Amphibia, anura)

Aplicando métodos citoquímicos para carboidratos nas glândulas "tubuloalveolares", localizadas na regiao posterior da lingua de Bufo ictericus, foi possível caracterizar como mucina neutra o produto de secreçao dos mucocitos destas glândulas.

Cytochemical variations of human von Ebner's glands

Se analizaron las glándulas de von Ebner humanas durante el desarrollo fetal, infantil y adulto, con el propósito de observar posibles cambios citoquímicos en relación con la edad. Se utilizaron lenguas de fetos de 8 a 38 semanas, muestras de la V lingual de niños recién nacidos, de 8 a 14 años, y adultos de 20 años en adelante, empleándose H/E, y técnicas para muco sustancias (PAS, PAS/amilasa, PAS/sialidasa, metanamina/plata, Alcian blue a pH 2,5 y 1,0 Azul de toluidina a pH 3,8). Entre las 16 y 20 semanas se identificaron acinosa y conductos en formación, estando el parénquima completamente desarrollado a las 24 semanas. Las glándulas de recién nacidos presentaban morfológicamente el aspecto de acinos serosos basófilos y periodatonegativos. En el niño las células secretoras contenían gránulos apicales PAS positivos sensibles a la sialidasa, periodatorreactivos y ligeramente metacromáticos. Estas características se acrecentaron con la edad. En el adulto el citoplasma apical de los adenómeros y el contenido luminal eran además alcianófilos. Las variaciones citoquímicas observadas indican que las glándulas de von Ebner humanas están constituidas por células de tipo seromucoso que contienen sialomucinas y sulfomucinas

Estudo histoquímico das glândulas salivares linguais posteriores do Coendu Villosus (Mammalia, Rodentia) / Histochemical study of the posterior lingual salivary glands of Coendu Villosus (Mammalia, Rodentia)

Os autores estudaram com métodos histoquímicos, a natureza do material elaborado pelas glândulas linguais posteriores de Coendu Villosus. Com base nos resultados obtidos, foi possível concluir que: 1) o produto de secreção das glândulas de Weber contém uma sulfosialomucina, fato que permite classificar essas glândulas como sendo do tipo mucoso; 2) o produto de secreção das glândulas de Von Ebner contém uma glico-proteína, tipo sero-mucoso