PI3K/Akt Signaling Involved with Osteoinductive Effects of Achatina fulica Mucus

Abstract Mesenchymal stem cells and osteoblasts play important roles in bone formation. Achatina fulica mucus presented the property of osteoinduction. This study aimed to examine the effects of A. fulica mucus on human mesenchymal stem cell (hMSC) and human fetal osteoblastic cell line (HFOB) differentiation. The integrated effects of A. fulica mucus and polycaprolactone (PCL) on the differentiation of hMSCs were tested. The cell viability of hMSCs treated with A. fulica mucus was investigated by the MTT assay. The cell mineralization was observed by Alizarin Red S staining, the gene expression was investigated using RT-PCR, and the PI3K activation was studied using flow cytometry. The results indicated that A. fulica mucus induced osteogenic differentiation in hMSCs and HFOBs by upregulation of the osteogenic markers; osteopontin (OPN) and osteocalcin (OCN). The results of the Alizarin Red S staining indicated that A. fulica mucus supported mineralization in both hMSCs and HFOBs. The hMSCs cultured on PCL supplemented with A. fulica mucus showed significantly increased RUNX2 and OPN expressions. A. fulica mucus was observed to increase PI3K activation in hMSCs. The findings of this study suggested that A. fulica mucus and biomaterials could be applied together for use in bone regeneration in the future.

The effects of smoking and smoking cessation on nasal mucociliary clearance, mucus properties and inflammation

OBJECTIVE: The aim of the present study was to assess nasal mucociliary clearance, mucus properties and inflammation in smokers and subjects enrolled in a Smoking Cessation Program (referred to as quitters). METHOD: A total of 33 subjects with a median (IQR) smoking history of 34 (20-58) pack years were examined for nasal mucociliary clearance using a saccharine transit test, mucus properties using contact angle and sneeze clearability tests, and quantification of inflammatory and epithelial cells, IL-6 and IL-8 concentrations in nasal lavage fluid. Twenty quitters (mean age: 51 years, 9 male) were assessed at baseline, 1 month, 3 months and 12 months after smoking cessation, and 13 smokers (mean age: 52 years, 6 male) were assessed at baseline and after 12 months. Clinicaltrials.gov: NCT02136550. RESULTS: Smokers and quitters showed similar demographic characteristics and morbidities. At baseline, all subjects showed impaired nasal mucociliary clearance (mean 17.6 min), although 63% and 85% of the quitters demonstrated significant nasal mucociliary clearance improvement at 1 month and 12 months, respectively. At 12 months, quitters also showed mucus sneeze clearability improvement (∼26%), an increased number of macrophages (2-fold) and no changes in mucus contact angle or cytokine concentrations. CONCLUSION: This study showed that smoking cessation induced early improvements in nasal mucociliary clearance independent of mucus properties and inflammation. Changes in mucus properties were observed after only 12 months of smoking cessation.

Chemical composition modulates the adverse effects of particles on the mucociliary epithelium

OBJECTIVE:We compared the adverse effects of two types of real ambient particles; i.e., total suspended particles from an electrostatic precipitator of a steel mill and fine air particles from an urban ambient particulate matter of 2.5 µm, on mucociliary clearance.METHOD:Mucociliary function was quantified by mucociliary transport, ciliary beating frequency and the amount of acid and neutral mucous in epithelial cells through morphometry of frog palate preparations. The palates were immersed in one of the following solutions: total suspended particles (0.1 mg/mL), particulate matter 2.5 µm 0.1 mg/mL (PM0.1) or 3.0 mg/mL (PM3.0) and amphibian Ringer’s solution (control). Particle chemical compositions were determined by X-ray fluorescence and gas chromatography/mass spectrometry.RESULTS:Exposure to total suspended particles and PM3.0 decreased mucociliary transport. Ciliary beating frequency was diminished by total suspended particles at all times during exposure, while particulate matter of 2.5 µm did not elicit changes. Particulate matter of 2.5 µm reduced epithelial mucous and epithelium thickness, while total suspended particles behaved similarly to the control group. Total suspended particles exhibited a predominance of Fe and no organic compounds, while the particulate matter 2.5 µm contained predominant amounts of S, Fe, Si and, to a lesser extent, Cu, Ni, V, Zn and organic compounds.CONCLUSION:Our results showed that different compositions of particles induced different airway epithelial responses, emphasizing that knowledge of their individual characteristics may help to establish policies aimed at controlling air pollution.

Assessment of antimicrobial activity and healing potential of mucous secretion of Achatina fulica / Evaluación de la actividad antimicrobiana y la cicatrización potencial de la secreción mucosa de Achatina fulica

Achatina fulica's secretion has been recently related to antibacterial, antifungal, and cicatricial properties, and it is influenced by the kind of food offered. Therefore, this study investigated the healing potential of dressing films based on mucous secretion of Achatina fulica. Thus, surgical wounds performed in black wistar rats were dressed with films based on collagen, and on mucous secretion of A. fulica fed with Lactuca sativa; undresses worked as wounds control. After 3, 7, 14 and 21 days the animals were euthanized, and the wounds were microscopically evaluated. On the 3rd and 7th days dressing films based on mucus provided acceleration of the formation maturation of granulation tissue, better epithelization rates, and more rapid replacement of type III for type I collagen fibers. On the 14th and 21st days, these dressings induced more intense deposition and better architectural disposition of type I collagen fibers, and hastened the regeneration of cutaneous phaneros. Dressings obtained from A. fulica fed with Lactuca sativa provided the most expressive results. This study suggests that films produced with mucous secretion of A. fulica can be successfully employed as wound dressing, particularly if snails are fed with Lactuca sativa.

A la secreción mucoproteica del Achatina fulica se le han asignado propiedades antibacterianas, antifúngicas y cicatriciales que pueden estar influidas por el tipo de alimento que se ofrece a este especimen. Este estudio investigó el potencial curativo de películas basadas en la secreción mucosa de Achatina fulica alimentaba con diferentes tipos de plantas. Fueron tratadas heridas provocadas en el dorso de ratas Wistar con películas de colágeno y películas realizadas a partir de la secreción mucosa de A. fulica, alimentados con plantas de lechuga (L. sativa), utilizando como parámetro de comparación la curación del grupo de control. Después de 3, 7, 14 y 21 días los animales fueron sacrificados y las heridas fueron evaluadas microscópicamente. En los días 3 y 7, las heridas tratadas con moco mostraron mejores tasas de formación y maduración del tejido de granulación, epitelización, y más rápido recambio de colágeno tipos III y I. A los días 14 y 21, hubo una intensa deposición del colágeno tipo I y aceleración en la regeneración de la piel. Los resultados obtenidos a partir de A. fulica alimentados con plantas de lechuga (L. sativa) mostraron mejores resultados. Este estudio sugiere que las películas producidas con secreción mucosa de A. fulica pueden ser utilizadas con éxito como apósito, especialmente si los caracoles se alimentan con plantas de lechuga L. sativa.

Histochemistry of the mucus gland of Bombus morio Swedurus,1787 (Hymenoptera, Apidae)

The male mucus gland is present in all species of bees, exception for meliponines. In bees, the function of themucus glands is not certain so far and its contribution to form the female espermatecal fluid is not assured.With the aim to contribute to the knowledge of the mucus gland in bees we carried out a histochemical studyof the gland in mature, adult males of Bombus morio, focusing on the histochemical nature of the secretion.The males were collected around the forest fragments into UFSCar, Campus Sorocaba. The glands werefixed and analysed under routine microscopy and for histochemistry techniques: Periodic Acid Schiff (PAS),Bromophenol Blue (BB), Sudan Black (SB) and Critical Electrolyte Concentration (CEC) variant. The resultsshowed that the mucus gland is constituted by a pair of large, thick tubular structures, which presented theirdistal portion more dilated and corn shaped. The glandular cells are columnar and in the apical portion presentseveral apocrine vesicles being released into the gland lumen. The histochemistry showed that gland secretionis very complex and contains protein (BB), neutral polysaccharide (PAS), as well as lipid (SB). The luminalgland secretion presents a background of homogenous content, presenting little dark dots stained by alltechniques used for this work. The investigation of the nucleolar activity (CEC) showed that the mucus glandepithelium is evolved in protein synthesis, presenting nucleoli developed and much RNA in the cytoplasm.

O muco traqueobrônquico humano mantido em temperatura ambiente e suas propriedades físico-químicas / Physicochemical properties of human tracheobronchial sputum maintained at room temperature

OBJETIVO: Verificar a influência da permanência em temperatura ambiente na análise da transportabilidade por ação ciliar e por tosse e do ângulo de contato do muco traqueobrônquico. MÉTODOS: Foi coletado muco hialino de 30 indivíduos sem doença pulmonar, e purulento de vinte pacientes com bronquiectasia. As amostras foram analisadas logo após a coleta e novamente após 24 h. RESULTADOS: Para o muco purulento, após 24 h em temperatura ambiente, houve aumento no deslocamento por tosse (96 ± 50 vs. 118 ± 61 mm) e diminuição do ângulo de contato (32 ± 6 vs. 27 ± 6 graus) (p < 0,05). Para o muco hialino não houve alterações nas medidas analisadas. CONCLUSÃO: O muco traqueobrônquico hialino pode ser armazenado em temperatura ambiente por 24 h sem que haja alterações em sua transportabilidade por ação ciliar ou em seu ângulo de contato. Por outro lado, o muco purulento não deve permanecer em temperatura ambiente por muitas h para que não se altere seu ângulo de contato e sua transportabilidade por tosse.

OBJECTIVE: To evaluate the effect that maintaining tracheobronchial sputum at room temperature has on the analysis of ciliary transport and cough, as well as on the contact angle. METHODS: Hyaline sputum was collected from 30 individuals without pulmonary diseases, and purulent sputum was collected from patients with bronchiectasis. The samples were analyzed immediately after collection and again after 24 h. RESULTS: After 24 h at room temperature, the purulent sputum presented an increase in cough-induced dislodgment (96 ± 50 vs. 118 ± 61 mm) and a decrease in the contact angle (32 ± 6 vs. 27 ± 6 degrees) (p < 0.05). For the hyaline sputum, there were no alterations in the parameters analyzed. CONCLUSION: Hyaline tracheobronchial sputum can be stored in room temperature for 24 h without presenting alterations in ciliary transport or contact angle. However, purulent sputum should not be stored at room temperature for many hours, since ciliary transport and contact angle might be altered as a result.

Morphology and protein patterns of honey bee drone accessory glands

We used light and transmission electron microscopy to examine the morphology of the accessory glands of immature and mature adult males of Apis mellifera L. We also made an electrophoretic analysis of the protein content of the mature gland. The glands of the immature male actively secrete a mucous substance that can be seen in the lumen of the gland of the mature male. This secretion stains with mercury bromophenol blue and with periodic acid-Schiff reaction, which stain glyconjugates. The protein content was higher in the lumen secretion than in the gland wall extracts. The electrophoresis patterns of the wall extracts were different from those of the secretion found in the gland lumen.

Morfologia e histoquímica de glicogênio e mucossubstâncias na próstata e glândulas bulbo-uretrais de Marmosa cinerea Temminck, 1824 (Marsupialia - Didelphidae) / Morphology, glycogen and mucosubstances histochemistry of the prostate and bulbo-urethral glands of the Marmosa cinerea Temminck, 1824 (Marsupialia - Didelphidae)

Sao descritos os aspectos histológicos e a distribuiçao de glicogênio e mucossubstâncias na próstata e nos três pares de glândulas bulbo-uretrais (Cowper) de Marmosa cinerea, marsupial Didelphidae com ampla distribuiçao geográfica no Brasil. Os três segmentos prostáticos, cranial, médio e caudal, apresentam túbulos secretores disseminados na mucosa uretral e sao divididos em zonas externa, média e interna, de acordo com as características morfológicas e tintoriais do epitélio secretor. No segmento cranial, o epitélio secreta glicogênio e mucossubstâncias neutras. No segmento médio sao produzidas mucossubstâncias neutras e sialomucinas, sendo sugerido um mecanismo apócrino de secreçao neste segmento. O epitélio do segmento caudal produz glicogênio, mucossubstâncias neutras e proteínas. O par lateral de bulbo-uretrais é formado por túbulos secretores longos e ramificados, que produzem mucossubstâncias neutras. O par intermédio constitui-se de túbulo-ácinos revestidos por células com porçao apical expandida e que secretam mucossubstâncias neutras, sialomucinas e sulfomucinas. Os túbulo-ácinos do par medial sao tortuosos e suas células produzem mucossubstâncias neutras.

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.