Bioconjugation of carbohydrates to gelatin sponges promoting 3D cell cultures

Gelatin sponges are widely employed as hemostatic agents, and are gaining increasing interest as 3D scaffolds for tissue engineering. To broaden their possible application in the field of tissue engineering, a straightforward synthetic protocol able to anchor the disaccharides, maltose and lactose, for specific cell interactions was developed. A high conjugation yield was confirmed by 1H-NMR and FT-IR spectroscopy, and the morphology of the resulting decorated sponges was characterized by SEM. After the crosslinking reaction, the sponges preserve their porous structure as ascertained by SEM. Finally, HepG2 cells cultured on the decorated gelatin sponges show high viability and significant differences in the cellular morphology as a function of the conjugated disaccharide. More spherical morphologies are observed when cultured on maltose-conjugated gelatin sponges, while a more flattened aspect is discerned when cultured onto lactose-conjugated gelatin sponges. Considering the increasing interest in small-sized carbohydrates as signaling cues on biomaterial surfaces, systematic studies on how small carbohydrates might influence cell adhesion and differentiation processes could take advantage of the described protocol

Interaction of Cisplatin with a CCHC Zinc Finger Motif

The interaction between cisplatin and an 18-residue CCHC zinc finger motif derived from a retroviral nucleocapsid protein (PyrZf18) has been studied using UV–visible, CD and 1H NMR spectroscopies and ESI-MS spectrometry. Cisplatin irreversibly blocks the cysteine zinc binding groups in the free peptide and is able to slowly eject zinc from the zinc–peptide complex. The observed end product of the reaction with cisplatin is a complex in which only one ammonia molecule is coordinated to platinum. After an initial binding with two cysteine residues and the formation of the (PyrZf18)– platinum–(NH3)2 complex, a release of one ammonia molecule occurs because of trans-labilization, and the third cysteine is coordinated, leading to a mixture of isomers and/or conformers of the (PyrZf18)–platinum–NH3 complex. The results are discussed with respect to the potential antiretroviral activity of platinum(II) compounds and to the possible interaction of cisplatin with the cellular nucleic acid binding proteins

Characterization of a Functional NTPDase in the Endoplasmic Reticulum of Rat Submandibular Salivary Gland

Nucleotidase activity and Ca-uptake were characterized in endoplasmic reticulum (ER) enriched rat submandibular gland (SMG) microsomal preparations. (i) Ca-uptake had characteristics of an ER Ca-ATPase. (ii) Nucleotidase activity was equally stimulated by calcium, magnesium and manganese, but with different Km values. (iii) Specific inhibitors of P-type Ca-ATPases were ineffective on nucleotidase activity, demonstrating that this activity was not related to calcium uptake and did not correspond to classical Ca2+ pumps. (iv) ATP and UTP were more efficient substrates, whereas ADP and UDP were hydrolyzed at significantly slower rate. (v) Nucleotidase activity was sensitive to mild detergent solubilization and insensitive to ionophore addition. (vi) Nucleotidase activity was strongly inhibited by suramin, a nucleoside triphosphate diphosphohydrolase (NTPDase) inhibitor. (vii) Nucleotidase activity exponentially diminished as function of time. All these observations are consistent with a NTPDase identity. The presence of a NTPDase was demonstrated by immunohistochemistry in rat SMG. Immunoreactivity was stronger in ductal cells than in mucous and serous acini. Although this enzyme was observed in the plasma membrane, colocalization with the ER marker calnexin revealed a specific subcellular localization in this organelle of all three types of cell. The putative function of this NTPDase activity in salivary glands is discussed.Fil: Ostuni, M. A.. Inserm; Francia. Université Paris; Francia. Universidad de Buenos Aires. Facultad de Odontología; ArgentinaFil: Egido, P.. Universidad de Buenos Aires. Facultad de Odontología; ArgentinaFil: Peranzi, G.. Inserm; Francia. Université Paris; FranciaFil: Alonso, Guillermo Luis. Universidad de Buenos Aires. Facultad de Odontología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Lacapere, J. J.. Inserm; Francia. Université Paris; FranciaFil: Gonzalez, D. A.. Inserm; Francia. Université Paris; Franci

A Pilot Study on Biochemical Profile of Follicular Fluid in Breast Cancer Patients

Breast cancer (BC) is the most common type of cancer among women in almost all countries worldwide and is one of the oncological pathologies for which is indicated fertility preservation, a type of procedure used to help keep a person's ability to have children. Follicular fluid (FF) is a major component of oocyte microenvironment, which is involved in oocyte growth, follicular maturation, and in communication between germ and somatic cells; furthermore, it accumulates all metabolites during oocytes growth. To obtain information about changes on fertility due to cancer, we aimed at investigating potential biomarkers to discriminate between FF samples obtained from 16 BC patients and 10 healthy women undergoing in vitro fertilization treatments. An NMR-based metabolomics approach was performed to investigate the FF metabolic profiles; ELISA and western blotting assays were used to investigate protein markers of oxidative and inflammatory stress, which are processes closely related to cancer. Our results seem to suggest that FFs of BC women display some significant metabolic alterations in comparison to healthy controls, and these variations are also related with tumor staging

Phytochemical profile of capsicum annuum l. Cv senise, incorporation into liposomes, and evaluation of cellular antioxidant activity

Overproduction of oxidants in the human body is responsible for oxidative stress, which is associated with several diseases. High intake of vegetables and fruits can reduce the risk of chronic diseases, as they are sources of bioactive compounds capable of contrasting the free radical effects involved in cancer, obesity, diabetes, and neurodegenerative and cardiovascular diseases. Capsicum annuum L. cv Senise is a sweet pepper that is grown in the Basilicata region (Italy). It is an important source of polyphenols, carotenoids, and capsinoids and can play a key role in human health. In this study, an ethanol extract was obtained from C. annuum dried peppers and the analysis of the phytochemical composition was performed by LC-ESI/LTQ Orbitrap/MS. The extract was incorporated into liposomes, which showed small size (~80 nm), good homogeneity, negative surface charge, and good stability in storage. The biological activity of the extract was evaluated in the human hepatoma (HepG2) cell line, used as model cells. The extract showed no cytotoxic activity and reduced the intracellular reactive oxygen species (ROS) level in stressed cells. The antioxidant activity was further improved when the extract was loaded into liposomes. Moreover, the extract promoted the expression of endogenous antioxidants, such as catalase, superoxide dismutase, and glutathione peroxidase through the Nrf-2 pathway evaluated by RT-PCR

Imaging in major salivary gland diseases

Most of the salivary glands diseases are characterized only by a few distinct clinical patterns. Medical history and clinical examination are still considered of great relevance. However, in order to obtaine a definite diagnosis, imaging techniques are required in most of the cases. Salivary glands ultrasonography (US) is the technique to be used as the first because US can easily differentiate calculosis, inflammatory diseases and tumors. Sonography is also frequently needed to perform needle aspiration or biopsy (FNAC). Sialography should be used essentially for assessing chronic sialoadenitis as well as Sjogren's syndrome. At present, Magnetic Resonance sialography should be preferred because of the greater sensibility in diagnosing inflammatory diseases of the salivary glands. It allows to evaluate both intraglandular oedema and nodules, so that incannulation of the salivary duct is not required. Computer Tomography (CT) and Magnetic Resonance imaging (MR) are useful when neoplasm are suspected, particularly if deep areas of the gland, which cannot be visualized by US, are involved. Sequential scintigraphy is currently employed for assessing the functional status of all the 4 major salivary glands and evaluating the chronic evolution of glandular damage

Electrokinetic remediation of metal-polluted marine sediments: experimental investigation for plant design

This paper presents the results of an extensive set of laboratory experiments performed to design a demonstrative electrokinetic plant for extracting heavy metals from marine sediments dredged from the Livorno marine harbour. The investigated sediments displayed a high salinity, a high acid neutralization capacity, a low electrical resistivity (0.5 Ωm), a high alkalinity (pH ≈ 8) and a large fraction of fine particles. The target metals were Cd, Cr, Cu, Ni, Pb and Zn at relatively weak and inhomogeneous concentrations with high non-mobile fractions. After an accurate characterization, several screening and full electrokinetic tests were performed using cells of two different sizes, several conditioning agents (HNO3, HCl, H2SO4, citric acid, oxalic acid, ascorbic acid, EDTA), different applied current intensities and durations. The tests highlighted the need for long treatment times in order to obtain a significant pH reduction, with some appreciable metal removal being attained only after several weeks. The best results were obtained with strong acids used as the conditioning agents, with significant specific effects of each acid, including pronounced resistivity increase (from 0.5 up to 10 Ωm) and a high electroosmotic flow (EOF) with H2SO4, or a reversed EOF (electroendosmosis), and minor resistivity changes with HNO3. The use of the obtained data to design a demonstrative plant is also presented in the paper, with considerations on operating parameters such as energy and reagent consumption, characteristics of plant components and required safety measures. buffer capacity, conditioning agent

Phytochemical profile, antioxidant and antidiabetic activities of Adansonia digitata L. (baobab) from Mali, as source of health-promoting compounds

Background: Adansonia digitata L. (Malvaceae), also known as baobab, is a tree attracting recent interest especially due to the high nutritional value of the fruit pulp. However, few studies are reported on the secondary metabolite content, showing high variability due to geographic region. Methods: In this study, the chemical profiles of Malian commercial baobab fruits and leaves, focused on phenolic content, were investigated by HPLC coupled with a photodiode array (PDA)/UV and an electrospray ionization (ESI) mass spectrometer (MS) and gas chromatography (GC)/MS. In addition, the extracts of fruit pulps obtained from three different markets (Fruits 1, 2, and 3) were evaluated for their total phenolic content (TPC), antioxidant activity and α-glucosidase inhibition. Results: Baobab fruit pulps were found to be rich in procyanidins and flavonol glycosides, with tiliroside as the major constituent. The baobab leaves showed a similar profile respect to the fruits, but with more detected phenolics. All fruit pulp extracts exerted antioxidant activity (highest for Fruit 3) and higher α-glucosidase inhibition than acarbose used as standard. Conclusions: This study confirmed the variability of baobab with different origins and indicated Malian species baobab as promising source of health-promoting substances

Are the Follicular Fluid Characteristics of Recovered Coronavirus Disease 2019 Patients Different From Those of Vaccinated Women Approaching in vitro Fertilization?

The aim of this pilot study is to evaluate if SARS-CoV-2 infection or vaccination against SARS-CoV-2 infection induce observable metabolic effects in follicular fluid of women who are following in vitro fertilization (IVF) treatments. The possible impact of coronavirus disease 2019 (COVID-19) on fertility and IVF outcome is considered. We have selected for this study: six women vaccinated against SARS-CoV-2 infection, five recovered COVID-19 patients, and we used nine healthy women as the control group. At the time of oocytes retrieval from participants in the study, follicular fluids were collected and metabolomic analysis was performed by 1H NMR spectroscopy in combination with multivariate analysis to interpret the spectral data. The search for antibody positivity in the follicular fluid aspirates was also carried out, together with the western blotting analysis of some inflammatory proteins, interleukin-6, tumor necrosis factor α (TNFα), and the free radical scavenger superoxide dismutase 2. Higher levels of Ala and Pro together with lower levels of lipids and trimethylamine N-oxide (TMAO) were found in follicular fluids (FFs) of vaccinated women while lower levels of many metabolites were detected in FFs of recovered COVID patients. Expression level of TNF-α was significantly lower both in recovered COVID-19 patients and vaccinated women in comparison to healthy controls

Standardization of a simple method to study whole saliva: clinical use in different pathologies.

En este trabajo se describe la normatización de un método para determinar flujo salival en humanos utilizando saliva total obtenida del piso de la boca mediante un eyector dental descartable y una bomba de vacío (equipo dental). En este estudio se evaluaron 40 sujetos sanos de ambos sexos y 51 pacientes con diversas patologías (Síndrome de Sjögren, Disfunción Tiroidea, Diabetes Mellitus). Se demostró que el flujo salival basal era estable a partir de los primeros 5 minutos de colocado el eyector en la cavidad bucal. No se encontraron diferencias significativas en el flujo salival basal comparando los sexos, siendo independiente de la intensidad del vacío efectuado por la bomba. El flujo de saliva total estimulada fue determinado durante 3 minutos, luego de los primeros 5 minutos de colocado el eyector en la boca. El estímulo se efectuó adosando en la cara dorsal de la lengua discos de papel absorbente, embebidos en ácido cítrico al 2 %. El uso de este método en pacientes con Síndrome de Sjögren confirmó la reducción del flujo salival respecto a los sujetos sanos. Los pacientes hipotiroideos y con neuropatía diabética demostraron disminución del flujo salival.The present study describes a methodology to assess the salivary flow rate in humans. Whole saliva was obtained from the floor of the mouth with a plastic dental ejector and a vacuum pump. Forty healthy subjects of both sexes and 51 patients with different pathologies (Sjögren Syndrome, Thyroid Dysfunction, Diabetes Mellitus) were included in the study. It was demonstrated that basal salivary flow rate was stable five minutes after the insertion of the oral ejector. Salivary flow rate did not show significant differences between sexes and was independent of the negative pressure level of the vacuum pump. Stimulated salivary flow rate was quantified over a period of 3 minutes, starting 5 minutes after the introduction of the oral device. The stimulus was paper filter disks soaked in citric acid (2%) placed on the tongue dorsum. The use of this method confirmed the reduction of salivary flow rate in patients with Sjögren Syndrome. In addition, a significant reduction in salivary flow rate was observed in patients with primary thyroid insufficiency and peripheral neuropathy secondary to Diabetes Mellitus.Fil: Tumilasci, Omar Rene. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Fisiología; ArgentinaFil: Cardoso, Estela M. del Luján. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Contreras, Liliana Noemí. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Belforte, Juan Emilio. Universidad de Buenos Aires. Facultad de Odontología. Cátedra de Fisiología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Arregger, Alejandro Luis. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Investigaciones Médicas. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Investigaciones Médicas; ArgentinaFil: Ostuni, Mariano A.. Inserm; Franci