Cytoreductive surgery (CRS) plus hyperthermic intraperitoneal chemotherapy (HIPEC) vs CRS alone for treatment of endometrial cancer with peritoneal metastases: a multi-institutional study from PSOGI and BIG RENAPE groups.

OBJECTIVE: To investigate the benefit of cytoreductive surgery (CRS) combined with hyperthermic intraperitoneal chemotherapy (HIPEC) for the treatment of endometrial peritoneal carcinomatosis compared to CRS alone. METHODS: We conducted a retrospective multicentre study of patients from experienced centres in treating peritoneal malignancies from 2002 to 2015. Patients who underwent surgery for peritoneal evolution of endometrial cancer (EC) were included. Two groups of 30 women were matched and compared: "CRS + HIPEC" which used HIPEC after CRS, and "CRS only" which did not use HIPEC. We analysed clinical, pathologic and treatment data for patients with peritoneal metastases from EC. The outcome measures were morbidity, overall survival (OS), and progression-free survival (PFS). RESULTS: In "CRS plus HIPEC" group, 96.7% of women were treated for recurrence, while in "CRS only" 83.3 were treated for primary disease. There was no significant difference between Peritoneal Carcinomatosis Index at laparotomy or Completeness of Cytoreduction score. Grade III and IV complications rates did not significantly differ between "CRS plus HIPEC" group and "CRS only" group (20.7% vs 20.7%, p = 0.739). Survival analysis showed no statistical difference between both groups. Median OS time was 19.2 months in "CRS plus HIPEC" group and 29.7 months in "CRS only" group (p = 0.606). Median PFS survival time was 10.7 months in "CRS plus HIPEC" group and 13.1 months in "CRS only" group (p = 0.511). CONCLUSION: The use of HIPEC combined to CRS did not have any significance as regard the DFS and OS over CRS alone in patients with primary or recurrent peritoneal metastasis of endometrial cancer.

Mechanosensitive regulation of stanniocalcin-1 by zyxin and actin-myosin in human mesenchymal stromal cells.

Stanniocalcin-1 (STC1) secreted by mesenchymal stromal cells (MSCs) has anti-inflammatory functions, reduces apoptosis, and aids in angiogenesis, both in vitro and in vivo. However, little is known about the molecular mechanisms of its regulation. Here, we show that STC1 secretion is increased only under specific cell-stress conditions. We find that this is due to a change in actin stress fibers and actin-myosin tension. Abolishment of stress fibers by blebbistatin and knockdown of the focal adhesion protein zyxin leads to an increase in STC1 secretion. To also study this connection in 3D, where few focal adhesions and actin stress fibers are present, STC1 expression was analyzed in 3D alginate hydrogels and 3D electrospun scaffolds. Indeed, STC1 secretion was increased in these low cellular tension 3D environments. Together, our data show that STC1 does not directly respond to cell stress, but that it is regulated through mechanotransduction. This research takes a step forward in the fundamental understanding of STC1 regulation and can have implications for cell-based regenerative medicine, where cell survival, anti-inflammatory factors, and angiogenesis are critical.

Effect of liquid whey protein concentrate-based edible coating enriched with cinnamon carbon dioxide extract on the quality and shelf life of Eastern European curd cheese.

Fresh unripened curd cheese has long been a well-known Eastern European artisanal dairy product; however, due to possible cross-contamination from manual production steps, high moisture content (50-60%), and metabolic activity of present lactic acid bacteria, the shelf life of curd cheese is short (10-20 d). Therefore, the aim of this study was to improve the shelf life of Eastern European acid-curd cheese by applying an antimicrobial protein-based (5%, wt/wt) edible coating. The bioactive edible coating was produced from liquid whey protein concentrate (a cheese production byproduct) and fortified with 0.3% (wt/wt, solution basis) Chinese cinnamon bark (Cinnamomum cassia) CO2 extract. The effect of coating on the cheese was evaluated within package-free (group 1) and additionally vacuum packaged (group 2) conditions to represent types of cheeses sold by small and big scale manufacturers. The cheese samples were examined over 31 d of storage for changes of microbiological (total bacterial count, lactic acid bacteria, yeasts and molds, coliforms, enterobacteria, Staphylococcus spp.), physicochemical (pH, lactic acid, protein, fat, moisture, color change, rheological, and sensory properties). The controlled experiment revealed that in group 1, applied coating affected appearance and color by preserving moisture and decreasing growth of yeasts and molds during prolonged package-free cheese storage. In group 2, coating did not affect moisture, color, or texture, but had a strong antimicrobial effect, decreasing the counts of yeasts and molds by 0.79 to 1.55 log cfu/g during 31 d of storage. In both groups, coating had no effect on pH, lactic acid, protein, and fat contents. Evaluated sensory properties (appearance, odor, taste, texture, and overall acceptability) of all samples were similar, indicating no effect of the coating on the flavor of curd cheese. The edible coating based on liquid whey protein concentrate with the incorporation of cinnamon extract was demonstrated to efficiently extend the shelf life of perishable fresh curd cheese, enhance its functional value, and contribute to a more sustainable production process.

Use of ultrafiltrated cow's whey for the production of whey cheese with Kefir or probiotics.

BACKGROUND: In Southern European countries, whey cheeses are normally produced with ovine or caprine whey. Cow's cheese whey can also be used, although the whey cheese yield is low (2-3%, w/v) which discourages its use. In the present study, bovine cheese whey was concentrated by ultrafiltration for the production of four types of whey cheeses (Requeijão): conventional, without any addition (WC); with 10% (w/w) addition of cream (WCC); with cream fermented with Kefir culture (WCCK); and with cream fermented with Bifidobacterium sp. culture (WCCBB12). RESULTS: Whey cheeses with cream presented lower protein content (330-360 g kg-1 , dry basis) and higher levels of total solids (220-250 g kg-1 ) and fat (300-330 g kg-1 , dry basis) than WC. C16:0 and C18:1 were the most abundant fatty acids present, with 31% and 38%, respectively. The small differences found concerning instrumental determination of colour and texture were not perceived by panelists. However, the presence of Kefir and probiotics decreased the elastic modulus (G') of the samples, as well as their viscosity. Fermentation with Kefir presented the highest counts of lactic acid bacteria (7 logUFC g-1 ). However, after 14 days of refrigerated storage, the counts of yeasts and moulds reached 6 logUFC g-1 in all products, indicating the need for appropriate packaging solutions. CONCLUSION: Ultrafiltration of bovine whey allows for the efficient production of bovine whey cheeses. The addition of cream fermented with Kefir or BB12 appears to be an efficient methodology to incorporate Kefir or probiotic bacteria in Requeijão, improving its nutritional and sensory characteristics, alongside the potential for the extension of its shelf-life. © 2020 Society of Chemical Industry.

Characteristics of lactose-free frozen yogurt with κ-carrageenan and corn starch as stabilizers.

Frozen yogurt is a type of dairy product that is considered to be a more healthful alternative to conventional ice cream due to its lower fat content and the presence of viable lactic acid bacteria. Lactose-free products are a growing trend in the dairy industry, and lactose-free yogurts and ice creams can both be found on the market. However, lactose-free frozen yogurt has not yet reached the market. In this study, we aimed to investigate the effect of adding κ-carrageenan (0.05, 0.1, and 0.15%) and corn starch (1, 2, and 3%) on acidity, texture, viscosity, overrun, melting properties, color attributes, and sensory characteristics of lactose-free frozen yogurts. Lactose was reduced by enzymatic hydrolysis during the fermentation process. The effectiveness of the hydrolysis was measured by HPLC, and lactose was reduced to 0.05% after 80 min of incubation with the enzyme. The addition of stabilizers did not change overrun and melting properties of frozen yogurt, but it did affect pH, titratable acidity, and color parameters. The product with 0.15% κ-carrageenan had the highest hardness and stickiness values. Moreover, κ-carrageenan had a positive effect on sensory attractiveness of lactose-free frozen yogurt, and it reduced the coarse texture in comparison with the control without stabilizers. A lactose-free frozen yogurt with good quality and nutritional characteristics was produced, particularly with the use of κ-carrageenan as stabilizer.

Liquid Whey Protein Concentrates Produced by Ultrafiltration as Primary Raw Materials for ThermalDairy Gels.

The aim of this work is to study the gelation properties of liquid whey protein concentrates (LWPC) produced by ultrafiltration (UF) as raw material for thermally induced gels intended for food applications. LWPC thermal gelation was performed using different types of LWPC (non--defatted, defatted and diafiltered) of different protein mass fractions and pH. Most of the produced gels showed viscoelastic behaviour. Non-defatted LWPC gave stronger heat-induced gels with a more cohesive microstructure, a higher water holding capacity and also higher elastic modulus (G') and viscous modulus (G''). Gel properties were not improved in products with lower content of non-protein compounds. As expected, the increase in protein mass fraction positively influences protein interactions. However, the pH is responsible for the equilibrium between attraction and repulsion forces in the gel components that influence gel hardness and water holding capacity.

Engineering a cardosin B-derived rennet for sheep and goat cheese manufacture.

Different sheep and goat cheeses with world-renowned excellence are produced using aqueous extracts of Cynara cardunculus flowers as coagulants. However, the use of this vegetable rennet is mostly limited to artisanal scale production, and no effective solutions to large-scale industrial applications have been reported so far. In this sense, the development of a synthetic rennet based on the most abundant cardoon milk-clotting enzymes (cardosins) would emerge as a solution for scalability of production and for application of these proteases as alternative rennets in dairy industry. In this work, we report the development of a new cardosin B-derived rennet produced in the generally regarded as safe (GRAS) yeast Kluyveromyces lactis. Using a stepwise optimization strategy-consisting of culture media screening, complemented with a protein engineering approach with removal of the plant-specific domain, and a codon optimization step-we successfully improved cardosin B production yield (35×) in K. lactis. We demonstrated that the secreted enzyme displays similar proteolytic properties, such as casein digestion profiles as well as optimum pH (pH 4.5) and temperature (40 °C), with those of native cardosin B. From this optimization process resulted the rennet preparation Vegetable Rennet (VRen), requiring no downstream protein purification steps. The effectiveness of VRen in cheese production was demonstrated by manufacturing sheep, goat, and cow cheeses. Interestingly, the use of VRen resulted in a higher cheese yield for all three types of cheese when compared with synthetic chymosin. Altogether, these results clearly position VRen as an alternative/innovative coagulant for the cheese-making industry.

Novel Functional Whey-Based Drinks with Great Potential in the Dairy Industry.

This work focuses on the production of liquid whey protein concentrates by ultrafiltration followed by thermal denaturation and homogenization of the ultrafiltrated concentrate, as well as on the production of ultrafiltrated permeates concentrated by reverse osmosis. Kefir grains (fresh and thawed) and/or commercial probiotic bacteria were inoculated in both liquid whey protein concentrates and concentrated ultrafiltrated permeates and grown at 25 °C for 24 h for the manufacture of fermented drinks. The physicochemical characterization (pH, titratable acidity, viscosity, and content of total solids, ash, fat and proteins) of the obtained drinks was then assessed and compared. Enumeration of viable microorganisms was carried out immediately after inoculation (at 0 h), during the fermentation period (at 12 and 24 h) and during refrigerated storage (at 48, 168 and 336 h). The fermented drinks showed acceptable physicochemical and sensorial properties, and contained above 7 log CFU/mL of lactococci and lactobacilli and 6 log CFU/mL of yeasts after 14 days of refrigerated storage, which is in agreement with the standards required by international organizations like European Food Safety Authority (EFSA) and Food and Drug Administration (FDA) for products containing probiotics. In summary, the strategy developed in this work contributes to the expansion of the applications of products derived from whey fractionation for the design of novel functional foods.

Injectable MMP-sensitive alginate hydrogels as hMSC delivery systems.

Hydrogels with the potential to provide minimally invasive cell delivery represent a powerful tool for tissue-regeneration therapies. In this context, entrapped cells should be able to escape the matrix becoming more available to actively participate in the healing process. Here, we analyzed the performance of proteolytically degradable alginate hydrogels as vehicles for human mesenchymal stem cells (hMSC) transplantation. Alginate was modified with the matrix metalloproteinase (MMP)-sensitive peptide Pro-Val-Gly-Leu-Iso-Gly (PVGLIG), which did not promote dendritic cell maturation in vitro, neither free nor conjugated to alginate chains, indicating low immunogenicity. hMSC were entrapped within MMP-sensitive and MMP-insensitive alginate hydrogels, both containing cell-adhesion RGD peptides. Softer (2 wt % alginate) and stiffer (4 wt % alginate) matrices were tested. When embedded in a Matrigel layer, hMSC-laden MMP-sensitive alginate hydrogels promoted more extensive outward cell migration and invasion into the tissue mimic. In vivo, after 4 weeks of subcutaneous implantation in a xenograft mouse model, hMSC-laden MMP-sensitive alginate hydrogels showed higher degradation and host tissue invasion than their MMP-insensitive equivalents. In both cases, softer matrices degraded faster than stiffer ones. The transplanted hMSC were able to produce their own collagenous extracellular matrix, and were located not only inside the hydrogels, but also outside, integrated in the host tissue. In summary, injectable MMP-sensitive alginate hydrogels can act as localized depots of cells and confer protection to transplanted cells while facilitating tissue regeneration.

Performance of the Alice PDx Device With the Somnolyzer Automated Scoring Algorithm for the Diagnosis of Obstructive Sleep Apnea.

Objective Automated scoring of respiratory events could allow a swifter obstructive sleep apnea (OSA) identification. We assessed the accuracy of the Alice PDx device with the Somnolyzer automated scoring algorithm, compared to the manually reviewed scoring by a trained sleep technician, for the diagnosis of OSA. Methods A prospective study was conducted between March 2021 and March 2022 in Centro Hospitalar do Baixo Vouga, a level 2 hospital in Aveiro, Portugal. Patients with high pre-test probability for OSA performed a type III home sleep apnea testing with the Alice PDx device. Data were scored automatedly by the Sleepware G3 with the Somnolyzer digital system and manually by a trained sleep technician. Correlation and dependent t-tests were used. Sensitivity, specificity, positive predictive values (PPVs), negative predictive values (NPVs), and area under the receiver operating characteristic curve (AUROC) of automated scoring were calculated. Data were analyzed using the Stata Statistical Software (Release 17, StataCorp., 2023, College Station, TX: StataCorp LLC). Results In 150 participants (mean age 57.8 ± 13.9 years), the mean apnea-hypopnea index (AHI) was 21.9 ± 21.8 events/hour by manual scoring and 25.4 ± 21.6 events/hour by automated scoring. The mean difference was 3.4 ± 4.4 events/hour, and a strong, positive, linear correlation was found between the two scores (r = 0.98). At the altered AHI (AHI ≥ 5 events/hour), mild, moderate, and severe OSA, the automated scoring sensitivity/specificity values were 91.2%/100.0%, 80.0%/68.6%, 91.6%/41.9%, and 98.1%/80.9%, respectively. The PPVs/NPVs for the same categories were 100.0%/69.4%, 89.3%/51.1%, 79.7%/66.7%, and 91.8%/95.0%, respectively. Finally, the AUROC was 0.85, 0.70, 0.73, and 0.93, respectively. Conclusion The automated scoring obtained from the Alice PDx portable device, using Sleepware G3 with the Somnolyzer digital system, seems accurate enough to diagnose OSA and validate the initiation of PAP therapy in the correct clinical setting. Nevertheless, it does not replace manual reviewing by a trained sleep technician in the case of mild and moderate OSA, to obtain a correct severity classification. With this valuable time-saving tool, we expect to hasten OSA diagnosis and treatment and thus tackle the underdiagnosis problem.

Sheep's Second Cheese Whey Edible Coatings with Oregano and Clary Sage Essential Oils Used as Sustainable Packaging Material in Cheese.

Sheep's second cheese whey (SCW), the by-product resulting from whey cheese production, was used as a component of cheese coatings containing oregano (Origanum compactum) and clary sage (Salvia sclarea) essential oils (EOs). SCW powder was obtained by the ultrafiltration/diafiltration of SCW followed by reverse osmosis and freeze drying. The coatings were produced with a mixture of SCW and whey protein isolate (WPI) using glycerol as plasticizer. Model cheeses were produced with cow´s milk and those containing SCW:WPI coatings; those with and without EOs were compared to controls without coating and with a commercial coating containing natamycin. At the end of ripening (28 days), the cheeses containing EOs presented higher water activity (ca. 0.930) and moisture content, as well as lower titratable acidity. Concerning color parameters, significant differences were also observed between products and as a result of ripening time. However, the use of SCW:WPI coatings did not significantly influence the color parameters at the end of ripening. Regarding texture parameters, the cheeses containing SCW:WPI coatings presented significantly lower values for hardness, chewiness, and gumminess. Significant differences were also observed for all microbial groups evaluated either between products and as a result of ripening time. In all cases, lactobacilli and lactococci counts surpassed log 7-8 CFU/g, while the counts of yeasts and molds increased steadily from ca. log 3 to log 6 CFU/g. The lowest counts of yeasts and molds were observed in the samples containing natamycin, but nonsignificant differences between products were observed. In conclusion, SCW:WPI cheese coatings can successfully substitute commercial coatings with the advantage of being edible packaging materials manufactured with by-products.

Low Fat Yoghurts Produced with Different Protein Levels and Alternative Natural Sweeteners.

The food industry is looking for substitutes for sucrose in food items due to the excessive consumption of products with added sugar and the demand for healthier products. Alternative natural sweeteners can help achieve this goal. Different types of low-fat yoghurts (1% fat), with low-protein and high-protein levels (3% and 4.5-6.5% protein, respectively), were produced using alternative natural sweeteners. The low-protein yoghurts were made with stevia (0.03% w/w) or agave syrup (4.5% w/w). The high-protein yoghurts were made with stevia (0.04% w/w), xylitol (6% w/w) or honey (6% w/w). Sucrose (6% w/w) was used as a control in both trials. pH and titratable acidity, CIEL*a*b* color parameters, syneresis index, rheology and the texture profile of the low-fat yoghurts were evaluated over refrigerated storage. All products underwent sensory evaluation by an untrained panel. The high-protein yoghurts were found to be more acidic (>1% as lactic acid), had a lower syneresis index (between 2.1 and 16.2%) and a better consistency (stronger gel structure) than the low-protein yoghurts. In terms of rheological parameters, stevia-sweetened yoghurts scored higher than the other sweetened yoghurts, showing a better gel structure. The different sweeteners tested did not significantly affect the sensory properties of the yoghurts, although the high-protein yoghurts scored higher for most of the attributes evaluated. Overall, consumers preferred stevia-sweetened yoghurts to yoghurts sweetened with sucrose or agave for the low-protein yoghurts. Of the tested formulations, those containing high protein with the alternative natural sweetener xylitol received higher scores in all attributes. These results reveal the potential of the tested natural sweeteners as sucrose substitutes, while contributing to improving the nutritional value of yoghurts.

Whey Cheeses Containing Probiotic and Bioprotective Cultures Produced with Ultrafiltrated Cow's Whey.

Bovine whey cheese (WC) is a product from southern European countries that presents some challenges: its production process involves high energy inputs; the yield is low; and WC has a short shelf life. The application of ultrafiltration (UF) to bovine whey before manufacture of WC and the employment of protective cultures can reduce these disadvantages. The objective of this research was the production of whey cheeses using ultrafiltrated bovine cheese whey with added probiotics or probiotics plus protective cultures. Three types of WC were produced: control CW without any addition (C); CW with the addition of the probiotic Lactobacillus acidophilus (LA5); and CW with the addition of Lactobacillus acidophilus plus a protective culture containing Lacticaseibacillus paracasei and Lacticaseibacillus rhamnosus (LA5FQ4). The WCs were stored under refrigerated conditions for 28 days. The products with added cultures presented lower pH values and higher titratable acidities when compared to the control. Sample LA5 presented the lowest pH and the highest titratable acidity, while LA5FQ4 presented intermediate values. Slight differences were observed between products regarding color parameters, chiefly resulting from storage time. The samples with added cultures were firmer when compared to the control, with LA5 cheeses showing the highest values at the end of the storage. Lactic acid bacteria (LAB) counts were on the order of log 8-9 CFU/g for the products with added cultures. Lower levels of yeasts and molds were detected on the sample with the protective culture (LA5FQ4), so that by the end of storage they presented counts one log cycle lower than C and LA5. Hence, the beneficial impact of the protective culture on the shelf life of the product is evident. Regarding sensory evaluation, LA5FQ4 cheeses obtained the highest scores for all parameters evaluated. It can be concluded that the use of UF associated with the addition of protective cultures can be very useful to reduce the energy consumption of the manufacturing process, to prolong the shelf life of WC and to improve its sensory properties.

3D Niche-Inspired Scaffolds as a Stem Cell Delivery System for the Regeneration of the Osteochondral Interface.

The regeneration of the osteochondral unit represents a challenge due to the distinct cartilage and bone phases. Current strategies focus on the development of multiphasic scaffolds that recapitulate features of this complex unit and promote the differentiation of implanted bone-marrow derived stem cells (BMSCs). In doing so, challenges remain from the loss of stemness during in vitro expansion of the cells and the low control over stem cell activity at the interface with scaffolds in vitro and in vivo. Here, this work scaffolds inspired by the bone marrow niche that can recapitulate the natural healing process after injury. The construct comprises an internal depot of quiescent BMSCs, mimicking the bone marrow cavity, and an electrospun (ESP) capsule that "activates" the cells to migrate into an outer "differentiation-inducing" 3D printed unit functionalized with TGF-ß and BMP-2 peptides. In vitro, niche-inspired scaffolds retained a depot of nonproliferative cells capable of migrating and proliferating through the ESP capsule. Invasion of the 3D printed cavity results in location-specific cell differentiation, mineralization, secretion of alkaline phosphatase (ALP) and glycosaminoglycans (GAGs), and genetic upregulation of collagen II and collagen I. In vivo, niche-inspired scaffolds are biocompatible, promoted tissue formation in rat subcutaneous models, and regeneration of the osteochondral unit in rabbit models.

Application of Ultrafiltration to Produce Sheep's and Goat's Whey-Based Synbiotic Kefir Products.

Membrane filtration technologies are the best available tools to manage dairy byproducts such as cheese whey, allowing for the selective concentration of its specific components, namely proteins. Their acceptable costs and ease of operation make them suitable for application by small/medium-scale dairy plants. The aim of this work is the development of new synbiotic kefir products based on sheep and goat liquid whey concentrates (LWC) obtained by ultrafiltration. Four formulations for each LWC based on a commercial kefir starter or traditional kefir, without or with the addition of a probiotic culture, were produced. The physicochemical, microbiological, and sensory properties of the samples were determined. Membrane process parameters indicated that ultrafiltration can be applied for obtaining LWCs in small/medium scale dairy plants with high protein concentration (16.4% for sheep and 7.8% for goats). Sheep kefirs showed a solid-like texture while goat kefirs were liquid. All samples presented counts of lactic acid bacteria higher than log 7 CFU/mL, indicating the good adaptation of microorganisms to the matrixes. Further work must be undertaken in order to improve the acceptability of the products. It could be concluded that small/medium-scale dairy plants can use ultrafiltration equipment to valorize sheep's and goat's cheese whey-producing synbiotic kefirs.

Sheep's Butter and Correspondent Buttermilk Produced with Sweet Cream and Cream Fermented by Aromatic Starter, Kefir and Probiotic Culture.

Small ruminant dairy products are common in some Mediterranean countries, in the Middle East and Africa, and can play a particular role in the development of rural areas. Butter has been the object of few research studies aimed at evaluating its potential as a vehicle for probiotic microorganisms. Moreover, the recovery of fermented buttermilk with functional properties can be considered an excellent opportunity to value this dairy byproduct. Therefore, the purpose of the present work was to develop different sheep butters and respective buttermilks after cream fermentation by: (1) a mesophilic aromatic starter (A); (2) a kefir culture (K); and (3) a mixture of probiotic bacteria (P). The butters and buttermilk produced with fermented cream were compared with non-fermented sweet cream (S) butter or buttermilk, respectively, regarding their physicochemical, microbiological and sensory characteristics. The adjusted production (%, w/v) obtained for butter were: S (44.48%), A (36.82%), K (41.23%) and P (43.36%). S, A and K butters had higher solids, fat and ashes contents than P butter. The probiotic butter had a total fat of ca. 75% (w/w), below the legal limits, while all others had fat levels above 81.5%. In all samples, the pH decreased and the acidity increased over 90 days of refrigerated storage. These variations were more evident in the P butter, which agrees with the highest lactic acid bacteria counts found in this sample. Differences in color between samples and due to storage time were also observed. In general, the butter samples tended to become darker and yellower after the 60th day of storage. Texture analysis showed comparable results between samples and greater hardness was observed for the P butter, most probably due to its higher relative saturated fatty acids content (66.46% compared to 62−64% in S, A and K butters). Regarding rheological properties, all butters showed pseudoplastic behavior, but butter P had the lowest consistency index (249 kPa.sn−1). The probiotic butter and the corresponding buttermilk had viable cell counts greater than 7 Log CFU/g, indicating their suitability as probiotic carriers. All products were well accepted by consumers and small, but non-significant, differences (p > 0.05) were observed in relation to the sensory parameters evaluated. In general, it can be concluded that the use of adequate starter cultures can allow the production of innovative and potentially healthier products, alongside the valorization of dairy byproducts, improving the income of small-scale producers.

Evaluation of the Characteristics of Sheep's and Goat's Ice Cream, Produced with UF Concentrated Second Cheese Whey and Different Starter Cultures.

Second cheese whey (SCW) is the by-product resulting from the manufacture of whey cheeses. In the present work, sheep (S) and goat (G) SCW concentrated by ultrafiltration (UF) were used in the production of ice creams. Concentrated liquid SCW samples with inulin added as a prebiotic were fermented with yoghurt, kefir and probiotic commercial cultures before being frozen in a horizontal frozen yoghurt freezer. The physicochemical, microbiological and sensory properties of the products were evaluated over 120 days of frozen storage. The products presented significant differences regarding these properties, specifically the higher total solids and protein contents of sheep's ice creams, which were higher compared to their goat ice cream counterparts. Sheep's ice creams also presented higher hardness and complex viscosity, which increased with storage. These ice creams also presented higher overrun and lower meltdown rates. The color parameters of the ice creams showed significant differences between formulations resulting from storage time. In all cases, Lactobacilli sp. cell counts were higher than log 6 CFU/g at the first week of storage. In the case of sheep's ice creams these values were maintained or increased until the 30th day, but decreased until the 60th day. Lactococci sp. counts surpassed log 7 CFU/g in all products, and these values were maintained until the end of storage, except in the case of G-Yoghurt and G-Kefir. Concerning the products containing probiotics, the sum of Lactococci sp. and Lactobacilli sp. counts was of the order log 8-9 CFU/g until the 60th day of storage, indicating that the probiotic characteristics of ice creams were maintained for at least 2 months. All products were well accepted by the consumer panel. Sheep's SCW ice creams were better rated regarding aroma, taste and texture. However, only the ranking test was able to differentiate preferences among formulations.

A comparative study of mesenchymal stem cells cultured as cell-only aggregates and in encapsulated hydrogels.

There is increasing evidence that cells cultured in three-dimensional (3D) settings have superior performance compared to their traditional counterparts in monolayers. This has been attributed to cell-cell and cell-extracellular matrix interactions that more closely resemble the in vivo tissue architecture. The rapid adoption of 3D cell culture systems as experimental tools for diverse applications has not always been matched by an improved understanding of cell behavior in different 3D environments. Here, we studied human mesenchymal stem/stromal cells (hMSCs) as scaffold-free self-assembled aggregates of low and high cell number and compared them to cell-laden alginate hydrogels with and without arginine-glycine-aspartic acid peptides. We observed a significant decrease in the size of cell-only aggregates over 14 days in culture compared to the cells encapsulated in alginate hydrogels. Alginate hydrogels had persistently more living cells for a longer period (14 days) in culture as measured by total DNA content. Proliferation studies revealed that a weeklong culture of hMSCs in 3D culture, whether as aggregates or cell-laden alginate hydrogels, reduced their proliferation over time. Cell cycle analysis found no significant differences between days 1 and 7 for the different culture systems. The findings of this study improve our understanding of how aggregate cultures differ with or without a hydrogel carrier, and whether aggregation itself is important when it comes to the 3D culture of hMSCs.

Installation of click-type functional groups enable the creation of an additive manufactured construct for the osteochondral interface.

Melt extrusion-based additive manufacturing (AM) is often used to fabricate scaffolds for osteochondral (OC) regeneration. However, there are two shortcomings associated with this scaffold manufacturing technique for engineering of tissue interfaces: (a) most polymers used in the processing are bioinert, and (b) AM scaffolds often contain discrete (material) gradients accompanied with mechanically weak interfaces. The inability to mimic the gradual transition from cartilage to bone in OC tissue leads to poor scaffold performance and even failure. We hypothesized that introducing peptide gradients on the surface could gradually guide human mesenchymal stromal cell (hMSC) differentiation, from a chondrogenic towards on osteogenic phenotype. To work towards this goal, we initially manufactured poly(ϵ-caprolactone)-azide (PCLA) and PCL-maleimide (PCLM) scaffolds. The surface exposed click-type functional groups, with a surface concentration in the 102pmol cm-2regime, were used to introduce bone morphogenic protein-2 or transforming growth factor-beta binding peptide sequences to drive hMSC differentiation towards osteogenic or chondrogenic phenotypes, respectively. After 3 weeks of culture in chondrogenic medium, we observed differentiation towards hypertrophic chondrogenic phenotypes with expression of characteristic markers such as collagen X. In osteogenic medium, we observed the upregulation of mineralization markers. In basic media, the chondro-peptide displayed a minor effect on chondrogenesis, whereas the osteo-peptide did not affect osteogenesis. In a subcutaneous rat model, we observed a minimal foreign body response to the constructs, indicating biocompatibility. As proof-of-concept, we finally used a novel AM technology to showcase its potential to create continuous polymer gradients (PCLA and PCLM) across scaffolds. These scaffolds did not display delamination and were mechanically stronger compared to discrete gradient scaffolds. Due to the versatility of the orthogonal chemistry applied, this approach provides a general strategy for the field; we could anchor other tissue specific cues on the clickable groups, making these gradient scaffolds interesting for multiple interfacial tissue applications.

[Pre-Exposure Prophylaxis for Human Immunodeficiency Virus in the Medical Curricula in Portugal: A Cross-Sectional Analysis].

INTRODUCTION: Pre-exposure prophylaxis (PrEP) has gained relevance as a method of prevention for HIV in certain people and settings. Following the publication of the guideline on PrEP prescribing in Portugal, we aimed to assess the knowledge of Portuguese Medical Students about PrEP. MATERIAL AND METHODS: An online survey was sent to Medical students of Portuguese Medical Schools. We conducted a descriptive analysis of the results and an analytic cross-sectional study to identify factors associated with "knowing about PrEP", "having had one class about PrEP" and "identifying eligible groups correctly". RESULTS: Of the 796 students that responded to the survey, 64.6% were aware of what PrEP is. Of these, 34.44% acquired this knowledge during their training. Out of the total amount of respondents, 4.77% could identify correctly and completely the eligible groups for PrEP. As the training years went by, the probability of being aware of PrEP, having had one class about PrEP, and identifying the eligible groups correctly, increased. Of the sixth-year students, 43.48% had had one class about PrEP and among the students that were aware of PrEP, 28% identified what the eligible groups were. After adjusting for the school year, we found differences between Medical Schools regarding the outcomes. The association between the different ways of learning about PrEP and the ability to correctly identify eligible groups for PrEP was not statistically significant. CONCLUSION: The differences between Medical Schools could be harmonized through changes in the medical curricula that would allow this topic to be addressed more often.

Introdução: A profilaxia pré-exposição (PrEP) ganhou relevância como método de prevenção do VIH em determinados indivíduos e contextos. Após a entrada em vigor das normas para prescrição em Portugal, pretendemos aferir o conhecimento em relação à PrEP entre os estudantes de Medicina em Portugal.Material e Métodos: Foi enviado um questionário online aos estudantes de Medicina das escolas médicas portuguesas. Foi feita uma análise descritiva dos resultados e um estudo transversal analítico para identificar fatores associados a "conhecer a PrEP", "ter tido uma aula de PrEP", e "identificar grupos elegíveis corretamente".Resultados: Dos 796 estudantes que responderam, 64,6% sabiam o que era a PrEP. Destes, 34,44% obteve conhecimento sobre a mesma durante a sua formação. Entre os respondentes, 4,77% identificaram correta e completamente os grupos elegíveis. Com o avançar do ano letivo, a probabilidade de conhecer a PrEP, ter tido uma aula de PrEP e identificar os grupos corretamente aumentava. No sexto ano, 43,48% tinham tido uma aula sobre PrEP e entre os que conheciam a PrEP, 28% identificaram os grupos elegíveis. Existem diferenças entre as escolas médicas após ajustamento para o ano letivo em relação aos resultados obtidos. A forma como se tomou conhecimento da PrEP não alterou de forma estatisticamente significativa a capacidade de identificar corretamente grupos elegíveisConclusão: As diferenças entre as escolas médicas poderão ser harmonizadas. Esta temática poderá ser reforçada nos respetivos currículos.