Development of bilayer tissue-engineered scaffolds: combination of 3D printing and electrospinning methodologies.

Although different fabrication methods and biomaterials are used in scaffold development, hydrogels and electrospun materials that provide the closest environment to the extracellular matrix have recently attracted considerable interest in tissue engineering applications. However, some of the limitations encountered in the application of these methods alone in scaffold fabrication have increased the tendency to use these methods together. In this study, a bilayer scaffold was developed using 3D-printed gelatin methacryloyl (GelMA) hydrogel containing ciprofloxacin (CIP) and electrospun polycaprolactone (PCL)-collagen (COL) patches. The bilayer scaffolds were characterized in terms of chemical, morphological, mechanical, swelling, and degradation properties; drug release, antibacterial properties, and cytocompatibility of the scaffolds were also studied. In conclusion, bilayer GelMA-CIP/PCL-COL scaffolds, which exhibit sufficient porosity, mechanical strength, and antibacterial properties and also support cell growth, are promising potential substitutes in tissue engineering applications.

Lower Urinary Tract Symptoms, Toileting Behaviors, and Pelvic Floor Health Knowledge in Younger and Older Women.

INTRODUCTION AND HYPOTHESIS: The primary aim was to examine the relationship between lower urinary tract symptoms (LUTS), toileting behaviors, and pelvic floor health knowledge in younger and older women. The secondary aim was to compare LUTS, toileting behaviors, and pelvic floor health knowledge in younger and older women. METHODS: The study included 85 younger women and 81 older women. LUTS were evaluated using the International Consultation on Incontinence Questionnaire-Female LUTS (ICIQ-FLUTS), toileting behaviors using Toileting Behavior-Women's Elimination Behaviors (TB-WEB), and pelvic floor health knowledge using the Pelvic Floor Health Knowledge Quiz (PFHKQ). RESULTS: In terms of LUTS, the most common symptoms in younger women are urgency (31.8%), bladder pain (28.2%), and frequency of incontinence episodes (35.3%), whereas in older women, the most common symptoms observed are urgency (64.2%), frequency of incontinence episodes (61.7%), intermittency (46.9%), urgency urinary incontinence (58%), and frequency of incontinence episodes (49.4%). Most unhealthy toileting behaviors correlated with LUTS in both groups (p < 0.05). Although a weak negative correlation was detected between ICIQ-FLUTS total score and PFHKQ function in younger women (p < 0.05), no correlation was found in older women (p < 0.05). Whereas premature voiding, straining during voiding, TB-WEB total score were higher in older women than in younger women (p < 0.05), delayed voiding, position preference for voiding, and PFHKQ function/dysfunction scores were higher in younger women than in older women (p < 0.05). CONCLUSIONS: Within the scope of preventive health services, it may be important to provide informative programs on healthy toileting behaviors and pelvic floor health to women of all ages in order to prevent LUTS.

Microbial membrane transport proteins and their biotechnological applications.

Because of the hydrophobic nature of the membrane lipid bilayer, the majority of the hydrophilic solutes require special transportation mechanisms for passing through the cell membrane. Integral membrane transport proteins (MTPs), which belong to the Major Intrinsic Protein Family, facilitate the transport of these solutes across cell membranes. MTPs including aquaporins and carrier proteins are transmembrane proteins spanning across the cell membrane. The easy handling of microorganisms enabled the discovery of a remarkable number of transport proteins specific to different substances. It has been realized that these transporters have very important roles in the survival of microorganisms, their pathogenesis, and antimicrobial resistance. Astonishing features related to the solute specificity of these proteins have led to the acceleration of the research on the discovery of their properties and the development of innovative products in which these unique properties are used or imitated. Studies on microbial MTPs range from the discovery and characterization of a novel transporter protein to the mining and screening of them in a large transporter library for particular functions, from simulations and modeling of specific transporters to the preparation of biomimetic synthetic materials for different purposes such as biosensors or filtration membranes. This review presents recent discoveries on microbial membrane transport proteins and focuses especially on formate nitrite transport proteins and aquaporins, and advances in their biotechnological applications.

Distinctive Processing Effects on Recovered Protein Isolates from Laurel (Bay) and Olive Leaves: A Comparative Study.

Although there is a well-known awareness of the nutritional potential of plant proteins, their utilization within food formulations is currently limited due to insufficient investigation of the functional properties or processing conditions. In this study, the protein contents of the remaining pulps of laurel (bay) (LL) and olive leaves (OL) after alcoholic washing (representing phenolic compound extraction), heat treatment (representing the usage of the leaves for tea brewing or as cooking aid), and deoiling process (representing oil extraction) were investigated. Bicinchoninic acid assay (BCA) indicated that the best protein yield was achieved with a direct isolation process after hexane oil removal. Both LL and OL isolates contained around 80% protein, but high temperature and alcohol content broke down the protein structure as well as decreased the final protein content (∼40%). Alcohol treatment appears to remove protein-bound phenols and increase fluorescence intensity in OL protein isolates while potentially causing structural alterations in LL proteins. In addition to a dramatic decrease in fluorescence intensity, the absolute zeta potentials of protein extracts of boiling OL and LL increased by 53 and 24%, respectively. The increased zeta potentials along with the decreased fluorescence intensity indicate the changes in the protein conformation and enhanced hydrophilicity of the protein structure, which can influence the functional properties of proteins. Protein extracts of deoiled LL had the highest ΔH value (180 mJ/mg), which is higher than other laurel and all olive protein samples. Laurel protein isolates became more thermally stable after hexane treatment. Moreover, the protein extracts after hexane treatment showed better emulsion capacity from both laurel (71.57%) and olive (61.87%). Water-binding capacity and thermal stability of the protein extracts from deoiled samples were higher than those of the other pretreatments, but the boiled samples showed higher oil-binding capacity due to protein denaturation. These findings indicate the importance of processing conditions in modulating protein properties for various applications.

Genome-Wide Identification and Expression Analysis of MTP (Metal Ion Transport Proteins) Genes in the Common Bean.

MTP/CDF carriers, called metal ion transport proteins, act as substrates for the transmission of micronutrients such as iron (Fe), zinc (Zn), and manganese (Mn) to membrane carriers in plants. In this study, genome-wide analysis of the MTP gene family in the common bean genome, expression analysis of the PvMTP4, PvMTP5, and PvMTP12 genes after Fe and Zn treatments, and the effects of Fe and Zn applications on iron and zinc content were investigated. This study used common bean genotypes assumed to have high or low Fe and Zn accumulation ability. PvMTP genes were defined as containing conserved catalytic domains with molecular weights and protein lengths ranging from 41.35 to 91.05 kDa and from 369 to 813 amino acids (aa), respectively. As a result of the phylogenetic analysis, three main clusters containing seven subgroups were formed. In this study, the first characterization of the MTP gene family of beans was performed, and the responses of three different PvMTP genes in the Zn-CDF group to Fe and Zn applications were revealed. The obtained findings are thought to constitute pioneering resources for future research on common bean biofortification studies, plant breeding related to Fe and Zn, and the functional characterization of the MTP gene family.

Development of plant-based biopolymer coatings for 3D cell culture: boron-silica-enriched quince seed mucilage nanocomposites.

Spheroid formation with spontaneous aggregation has captured interest in most cell culture studies due to its easy set-up and more reliable results. However, the economic and technical costs of the advanced systems and commercial ultra-low adhesive platforms have pushed researchers into pursuing alternatives. Nowadays, polymeric coatings, including poly-hydroxyethyl methacrylate and agar/agarose, are the commonly used polymers for non-adhesive plate fabrication, yet the costs and working solvent or heat-dependent preparation procedures maintain the need for the development of novel biomaterials. Here, we propose a greener and more economical approach for producing non-adherent surfaces and spheroid formation. For this, a plant waste-based biopolymer from quince fruit (Cydonia oblonga Miller, from Rosaceae family) seeds and boron-silica precursors were introduced. The unique water-holding capacity of quince seed mucilage (Q) was enriched with silanol and borate groups to form bioactive and hydrophilic nanocomposite overlays for spheroid studies. Moreover, 3D gel plates from the nanocomposite material were fabricated and tested in vitro as a proof-of-concept. The surface properties of coatings and the biochemical and mechanical properties of the nanocomposite materials were evaluated in-depth with techniques, and extra hydrophilic coatings were obtained. Three different cell lines were cultured on these nanocomposite surfaces, and spheroid formation with increased cellular viability was recorded on day 3 with a >200 µm spheroid size. Overall, Q-based nanocomposites are believed to be a fantastic alternative for non-adherent surface fabrication due to their low-cost, easy operation, and intrinsic hydration layer forming capacity with biocompatible nature in vitro.

Comparison of ion selectivities of nitrite channel NirC and water channel aquaporin.

nirC gene coding for the nitrite channel of E. coli K12 was cloned into the pET28a vector and expressed in E. coli BL21 cells. 28.5 kDa NirC monomer was purified from membrane components of E. coli. Selectivity of NirC for different ions including nitrite, nitrate, sulfate, formate, and acetate anions, and a divalent cation, magnesium, was compared with that of bacterial aquaporin from Halomonas elongata. Water and ion permeability values were determined by measuring the light scattering rates of proteoliposomes containing NirC and aquaporins during their water loss and gain. NirC shows a selective permeability to nitrite and is more resistant to the entry of other anions as compared to aquaporin. The single channel permeability of NirC for nitrite is about 10-fold that of a single aquaporin channel. Both aquaporin and NirC channel proteins were impermeable to MgCl2 and (NH4)2SO4 and their permeability to other tested ions was remarkably lower as compared to nitrite ions. The study also presents the 3D model and channel characteristics of NirC. The translocation channel of E. coli NirC is determined to be larger, and its length is shorter than aquaporin channels. Although the NirC channel throat is more hydrophobic than aquaporin, its water permeability is almost equal to that of aquaporin. The hydrophobic nature of the NirC channel might play an important role in the selective permeability of the channel for nitrite ions.

Effect of de-phenolization on protein-phenolic interactions of sunflower protein isolate.

Proteins and phenolic compounds are significant components of foods that can interact, and this interaction can impact the functional properties of proteins and the bioactivity of phenolic compounds. Sunflower meal, which has a high potential to be an important alternative protein source, contains phenolic compounds mostly bonded with proteins. In this study, the interaction between proteins and phenolic compounds which naturally exist in sunflower and prone to oxidation during alkaline treatment (for protein isolation) was investigated. There was a significant decrease up to 96.21% in the content of total phenolics by methanol washing. Chlorogenic acid, cryptochlorogenic acid and caffeic acid were detected in the phenolic extract obtained from sunflower protein isolate, and they exhibited different levels of reduction after methanol washing. For the total antioxidant capacity analysis, a decrease by 50% was observed after 4hwashing with methanol solution, and there was no significant decrease afterwards. In addition, the fluorescence intensity of sunflower protein was diminished with reduced washing time, which was mostly attributed to the protein-phenolic interaction. According to hydrodynamic parameters, the main force of the sunflower protein-phenolic complex formation was assumed to be hydrophobic attraction. The Stern-Volmer plot indicated that the main quenching mechanism was only static at all temperature conditions.

Nitrite is reduced by nitrite reductase NirB without small subunit NirD in Escherichia coli.

The assimilatory nitrite reductase enzyme NirB and small subunit NirD genes encoded in nir operon in Escherichia coli were cloned into the pET28a vector, and the recombinant enzyme was characterized for the first time. Docking of NirB with NirD, NADH, NO2-, NO3-, and CHO2- was performed using docking modeling programs. Methyl viologen and sodium dithionite were used as electron couples, and the amount of reduced nitrite was measured to calculate enzyme activity. NirB is the main enzyme and shows high activity with or without NirD. However, the inclusion of NirD into the enzyme solution at a ratio of 1NirD:2NirB resulted in 10% higher nitrite reductase activity. The enzyme tends to aggregate in the absence of ß-mercaptoethanol, which causes the conversion of tetrameric NirB to monomeric form, and the NirB enzyme shows its highest activity in monomeric form. The optimum temperature for enzyme activity was 37 °C and the optimum pH was found to be 7.0. Km and Vmax values of NirB were calculated as 9833 µM and 416.67 µmol NO2- reduced min-1 mg-1. Enzyme activity decreased by 55% and 50% in the presence of 100 mM nitrate and formate, respectively. The presence of 25 mM Cd2+ protected the enzyme at room temperature and the enzyme showed 10% higher activity in the presence of cadmium.

Chemistry of Protein-Phenolic Interactions Toward the Microbiota and Microbial Infections.

Along with health concerns, interest in plants as food and bioactive phytochemical sources has been increased in the last few decades. Phytochemicals as secondary plant metabolites have been the subject of many studies in different fields. Breakthrough for research interest on this topic is re-juvenilized with rising relevance in this global pandemics' era. The recent COVID-19 pandemic attracted the attention of people to viral infections and molecular mechanisms behind these infections. Thus, the core of the present review is the interaction of plant phytochemicals with proteins as these interactions can affect the functions of co-existing proteins, especially focusing on microbial proteins. To the best of our knowledge, there is no work covering the protein-phenolic interactions based on their effects on microbiota and microbial infections. The present review collects and defines the recent data, representing the interactions of phenolic compounds -primarily flavonoids and phenolic acids- with various proteins and explores how these molecular-level interactions account for the human health directly and/or indirectly, such as increased antioxidant properties and antimicrobial capabilities. Furthermore, it provides an insight about the further biological activities of interacted protein-phenolic structure from an antiviral activity perspective. The research on the protein-phenolic interaction mechanisms is of great value for guiding how to take advantage of synergistic effects of proteins and polyphenolics for future medical and nutritive approaches and related technologies.

Micropollutant removal capacity and stability of aquaporin incorporated biomimetic thin-film composite membranes.

Aquaporin incorporated nanofiltration membranes have high potential for future applications on separation processes. In this study, performance of biomimetic thin-film composite membranes containing Halomonas elongata and Escherichia coli aquaporins with different affinity tags for the removal of micropollutants was investigated.% rejection of the membranes for atrazine, terbutryn, triclosan, and diuron varied between 66.7% and 90.3% depending on the type of aquaporin and micropollutant. The highest removal rate was achieved with a membrane containing H. elongata aquaporin for atrazine and terbutryn which have methyl branching in their structure. Electrostatic interactions between micropollutants, thin-film layer of the membrane, and tags of aquaporins may also play important role in rejection of micropollutants. Stability experiments showed that biomimetic membranes can be used for six months period without a remarkable decrease in% rejection. Membrane used 24 times for atrazine removal for a year period lost most of its ability to repel atrazine.

Nano-hydroxyapatite incorporated quince seed mucilage bioscaffolds for osteogenic differentiation of human adipose-derived mesenchymal stem cells.

In this study, the therapeutic hydrocolloid quince seed mucilage (QSM) from Cydonia oblonga Miller fruit is enriched with needle-like nano-hydroxyapatite (nHAp) crystals to fabricate a novel biomimetic osteogenic bioscaffold. The molecular weight (Mw) of water-based extracted QSM was measured with GPC (8.67 × 105 g/mol), and the composite blend was prepared at a ratio of 1:1 (w/w) QSMaq and nHAp. The porous bioscaffolds were manufactured by the freeze-drying method, and evaluated in-depth with advanced analyses. The XRD, ATR-FTIR, SEM-EDX, and elemental mapping analyses revealed a uniform coated semi-crystalline structure with no covalent bindings between QSM and nHAp. Moreover, due to the hydrocolloid backbone, a supreme swelling ratio (w/w, 6523 ± 190%) with suitable pore size (208.12 ± 99.22 µm) for osteogenic development was obtained. Further, the cytocompatible bioscaffolds were evaluated for osteogenic differentiation in vitro using human adipose-derived mesenchymal stem cells (hAMSCs). The immuno/histochemical (I/HC) staining revealed that the cells with the spherical morphology invaded the pores of the prepared bioscaffolds. Also, relatively early up-regulated osteogenic markers were observed by the qRT-PCR analyses. Overall, it is believed that the QSM-nHAp bioscaffolds might be favorable in non-load bearing applications, especially in the cranio-maxillofacial region, due to their regenerative, bendable, and durable features.

Interactions of ß-Lactoglobulin with Bovine Submaxillary Mucin vs. Porcine Gastric Mucin: The Role of Hydrophobic and Hydrophilic Residues as Studied by Fluorescence Spectroscopy.

The aim of this study was to investigate binding interactions between ß-lactoglobulin (BLG) and two different mucins, bovine submaxillary mucins (BSM) and porcine gastric mucin (PGM), using intrinsic and extrinsic fluorescence spectroscopies. Intrinsic fluorescence spectra showed an enhanced decrease of fluorescence intensity of BLG at all pH conditions when BLG was mixed with PGM rather than with BSM. We propose that, unlike BSM, the tertiary structure of PGM changes and the hydrophobic regions are exposed at pH 3 due to protonation of negatively charged residues. Results suggest that PGM also facilitated the structural unfolding of BLG and its binding with PGM by a hydrophobic interaction, especially at acidic pH, which was further supported by extrinsic fluorescence spectroscopy. Hydrophobic interaction is suggested as the dominant interaction mechanism between BLG and PGM at pH 3, whereas electrostatic interaction is the dominant one between BLG and BSM.

From a plant secretion to the promising bone grafts: Cryogels of silicon-integrated quince seed mucilage by microwave-assisted sol-gel reaction.

Design and fabrication of biologically active cryogels using novel biopolymer(s) are still of great importance at regenerating bone defects such as traumatic bone injuries, maxillofacial surgery, osteomyelitis, and osteoporosis. Nowadays, plant mucilage, an herbal biomaterial, has been drawn attention by scientists due to their marvelous potential to fabricate 3-dimensional (3D) physical constructs for the field of regenerative medicine. Herein, a 3D cryogel from silicon-integrated quince seed mucilage (QSM) is constructed using microwave-assisted sol-gel reaction, characterized in-depth by attenuated total reflectance Fourier transform-infrared spectroscopy (ATR-FTIR), solid-state silicon cross-polarization magic-angle nuclear magnetic resonance (29Si-CP-MAS NMR), X-ray diffraction (XRD), thermogravimetric analysis (TGA), differential scanning calorimeter (DSC), micro-mechanical testing, porosity, and swelling tests, contact angle measurements, Brunauer-Emmet-Teller and Barret-Joyner-Halenda (BET-BJH) analysis, enzymatic biodegradation test, and field emission-scanning electron microscopy-energy dispersive X-ray spectroscopy (FE-SEM-EDX) mapping. The osteobiologic capacity of the cryogels is determined using human adipose-derived mesenchymal stem cells (hAMSCs) under in vitro conditions. Osteogenic differentiation of hAMSCs on both QSM and silica-modified QSM (Si-QSM) cryogels is analyzed by histochemistry, immunohistochemistry, and quantitative-real time (q-RT) PCR techniques. The results obtained from in vitro experiments demonstrate that the upregulation of osteogenesis-related genes in Si-QSM cryogels presents a stronger and earlier development over QSM cryogels throughout the culture period, which in turn reveals the great potential of this novel Si-incorporated QSM cryogels for bone tissue engineering applications.

Affinity tag effect on the salt rejection potential of Halomonas elongata aquaporin incorporated in thin film nanocomposite membrane.

In this study, effect of affinity tags, Histidine (His) and Glutathione-S-Transferase (GST), on the activity of halophilic aquaporin was analyzed. The gene coding for H. elongata aquaporin was cloned into pET28a vector and expressed in E. coli BL21 successfully. Stopped flow light scattering measurements showed that His-tagged aquaporin is functional. The difference in the filtration parameters caused by affinity tags were determined by using thin film composite nano-filtration (NFC) membranes prepared with the aquaporins. At 100 mM salt concentration, water permeability (L/m2.h) and the % salt rejection of NFC membranes produced with the His-tagged aquaporin was found to be higher than that of the membrane with GST-tagged aquaporin. Salt rejection of His-tagged aquaporin-membrane was found to be 53% with a lower solute permeability value (B). Use of short affinity tag (His tag) for cloning resulted in higher solute rejection ability of TFC membranes prepared with H. elongata aquaporins.

Validity and reliability of the Clinical Learning Environment, Supervision and Nurse Teacher (CLES+T), Turkish version1.

OBJECTIVE: A methodological type of study was conducted for the purpose of investigating the validity and reliability of the Turkish version of the Clinical Learning Environment, Supervision and Nurse Teacher (CLES+T) evaluation scale of the clinical learning environment of students, clinical nurses, and educators. METHODS: Sample was comprised of 602 Turkish nursing students with clinical practice experience at the hospital. The CLES+T, developed by Saarikoski, was used for data collection. Language equivalency, internal consistency, item-total correlation, and structure validity were conducted within the scope of the validity and reliability study on the CLES +T scale. RESULTS: It was determined that item-total correlations of four items were lower than 0.30, and those items were removed from the scale as a result of item analysis. The Cronbach's alpha value of the scale was 0.93-0.99; item total point correlations of the scale varied between 0.45 and 0.66; six factors were identified in the CLES+T factor analysis study, with a total variance explained by these six factors of 64%. CONCLUSION: According to the findings of the research, the CLES+T Turkish version was found to be a valid and reliable scale, which can be used to evaluate satisfaction of nursing students with their clinical education in Turkey.

Late-onset congenital adrenal hyperplasia with Cushing syndrome.

Although hirsutism is classically part of the clinical presentation of polycystic ovarian syndrome (PCOS), congenital adrenal hyperplasia and Cushing's syndrome (CS), CS associated with underlying late-onset congenital adrenal hyperplasia (LCAH) in an adult has not been previously reported. We herein present the case of a 25-year-old woman who was followed for PCOS for seven years. After undergoing detailed tests described within the text, she received the diagnosis of LCAH and was found to have point mutations. Interestingly, she later had diagnosis of endogenous CS that regressed folowing excision of an adrenal adenoma found on MRI. The present patient thus exhibited the coexistence of two paradoxical endocrine pathologies.