In vitro evaluation of the effect of yogurt acid whey fractions on iron bioavailability.

A side effect of the raised consumption of Greek yogurt is the generation of massive amounts of yogurt acid whey (YAW). The dairy industry has tried several methods for handling these quantities, which constitute an environmental problem. Although the protein content of YAW is relatively low, given the huge amounts of produced YAW, the final protein amount in the produced YAW should not be underestimated. Taking into consideration the increased interest for bioactive peptides and the increased demand for dietary proteins, combined with protein and peptides content of YAW, efforts should be made toward reintroducing the latter in the food supply chain. In this context and in view of the prevalent dietary iron deficiency problem, the objective of the present study was the investigation of YAW fractions' effect on Fe bioavailability. With this purpose, an in vitro digest approach, following the INFOGEST protocol, was coupled with the Caco2 cell model. To evaluate whether YAW digest fractions exert positive, negative or neutral effect on Fe bioavailability, they were compared with the ones derived from milk, a well-studied food in this context. Milk and YAW showed the same effectiveness on both Fe bioavailability and the expression of relative genes (DCYTB, DMT1, FPN1, and HEPH). Focusing further on YAW fractions, by comparison with their blank digest control counterparts, it resulted that YAW 3- to 10-kDa digests fraction had a superior effect over the 0- to 3-kDa fraction on Fe-uptake, which was accompanied by a similar effect on the expression of Fe metabolism-related genes (DCYTB, FPN1, and HEPH). Finally, although the 3- to 10-kDa fraction of bovine YAW digests resulted in a nonsignificant increased Fe uptake, compared with the ovine and caprine YAW, the expression of DCYTB and FPN1 genes underlined this difference by showing a similar pattern with statistically significant higher expression of bovine compared with ovine and bovine compared with both ovine and caprine, respectively. The present study deals with the novel concept that YAW may contain factors affecting Fe bioavailability. The results show that it does not exert any negative effect and support the extensive investigation for specific peptides with positive effect as well as that YAW proteins should be further assessed on the prospect that they can be used in human nutrition.

Genomic distribution and polymorphism of G-quadruplex motifs occupying ovine promoters and enhancers.

G-quadruplexes are non-canonical DNA structures that are formed in regions with short runs of guanines. During the last decade they have gained considerable attention due to their involvement in basic cellular processes, linking them to several physiological processes and pathological conditions. Regulation of gene transcription is among the crucial roles that G-quadruplexes play in the cells. Several ways in which these structures affect transcription have been described, both negatively and positively. Recently, G-quadruplexes have been shown to be implicated in the three-dimensional rearrangement of the chromosomes that enables the interaction of enhancers and gene promoters during regulation of transcription. Sheep is a species for which almost no G-quadruplex-related studies have been conducted and thus research on this species is kept out from the progress that has been made in the G-quadruplex field. In this context, we investigated the DNA sequences with potential to form G-quadruplexes (G4-motifs) in the ovine enhancers and promoters. We describe the distribution of G4-motifs within the regulatory regions which is shown to be enriched in G4-motifs in a way similar to other mammals. Furthermore, our data suggest that G4-motifs promote promoter-enhancer interactions in sheep. The single nucleotide polymorphisms colocalizing with promoter- and enhancer-associated ovine G4-motifs constitute a considerable pool of polymorphism and given the crucial role of these specific G4-motifs on regulation of transcription, we suggest this polymorphism as an interesting target for ovine genetic studies.

Genomic landscape, polymorphism and possible LINE-associated delivery of G-quadruplex motifs in the bovine genes.

G-Quadruplex structures are non-B DNA structures that occur in regions carrying short runs of guanines. They are implicated in several biological processes including transcription, translation, replication and telomere maintenance as well as in several pathological conditions like cancer and thus have gained the attention of the scientific community. The rise of the -omics era significantly affected the G-quadruplex research and the genome-wide characterization of G-Quadruplexes has been rendered a necessary first step towards applying genomics approaches for their study. While in human and several model organisms there is a considerable number of works studying genome-wide the DNA motifs with potential to form G-quadruplexes (G4-motifs), there is a total absence of any similar studies regarding livestock animals. The objectives of the present study were to provide a detailed characterization of the bovine genic G4-motifs' distribution and properties and to suggest a possible mechanism for the delivery of G4-motifs in the genes. Our data indicate that the distribution of G4-motifs within bovine genes and the annotation of said genes to Gene Ontology terms are similar to what is already shown for other organisms. By investigating their structural characteristics and polymorphism, it is obvious that the overall stability of the putative quadruplex structures is in line with the current notion in the G4 field. Similarly to human, the bovine G4-motifs are overrepresented in specific LINE repeat elements, the L1_BTs in the case of cattle. We highlight the potential role of these elements as vehicles for delivery of G4-motifs in the introns of the bovine genes. Lastly, it seems that a basis exists for connecting traits of agricultural importance to the genetic variation of G4-motifs, thus, the value of cattle as an interesting new model organism for G4-related genetic studies might be worth investigating.

Schizochytrium sp. Dietary supplementation modify Toll-like receptor 4 (TLR4) transcriptional regulation in monocytes and neutrophils of dairy goats.

Animals protect themselves against pathogens or abiotic factors by innate or adaptive mechanisms. Long-chain polyunsaturated fatty acids (ω3) of microalgae modify both human and mice' immune systems resulting in a beneficial balance between pro-inflammatory and anti-inflammatory pathways. However, scarce information exists on their impact on lactating animals' immunity. The objective of this study was to investigate the impact of dietary inclusion of Schizochytrium sp. (rich in docosapentaenoic and docosahexaenoic acid), on the expression of several genes involved in the innate immunity of goats. Twenty-four dairy goats were divided into four homogeneous sub-groups (n = 6). All goats were fed individually with alfalfa hay and concentrate. The concentrate of the control group (CON) had no microalgae while those of the treated groups were supplemented daily with 20 (ALG20), 40 (ALG40), and 60 (ALG60) g Schizochytrium sp. Monocytes and neutrophils were isolated from goats' blood in the 20th, 40th, and 60th days from the beginning of the experimental period. The relative transcript levels of TLR4, MYD88, MAPK, IRF3, IFNG, and pro-inflammatory cytokines (IL1B, IL2, IL8, TNF), and chemokines (CCL5 and CXCL16) were decreased in monocytes of microalgae treated goats compared to the CON. In contrast, MAPK and IL1B relative transcript levels were increased in neutrophils of ALG40 and ALG60 groups. In conclusion, the supplementation of goats' diet with 20 g Schizochytrium sp. resulted in a downregulation of the pro-inflammatory transcriptions, and following further research could be considered as a sustainable alternative strategy to improve immune function.

DNA G-quadruplexes: functional significance in plant and farm animal science.

G-quadruplexes (G4s) are non-canonical structures that can be formed in DNA and RNA sequences which carry four short runs of guanines. They are distributed in the whole genome but are enriched in gene promoter regions, gene UTRs and chromosome telomeres. The whole array of their functional roles is not fully explored yet but there is solid evidence supporting their implication in a number of processes like regulation of transcription, replication and telomere organization, among others. During the last decade, there is an increased research interest for G4s that has resulted in a better understanding of their role in several physiological and pathological conditions. On the other hand, these structures are poorly studied in plant species and animals of agricultural interest. Here, we summarize the current methods that are used for studying G4s, we review the studies concerning plants and farm animals and we discuss the advantages of a more thorough inclusion of G4s research in the agricultural sciences.

Effect of ethanol/TEOS ratios and amount of ammonia on the properties of copper-doped calcium silicate nanoceramics.

Calcium magnesium silicate glasses could be suggested for the synthesis of scaffolds for hard tissue regeneration, as they present a high residual glassy phase, high hardness values and hydroxyapatite-forming ability. The use of trace elements in the human body, such as Cu, could improve the biological performance of such glasses, as Cu is known to play a significant role in angiogenesis. Nano-bioceramics are preferable compared to their micro-scale counterparts, because of their increased surface area, which improves both mechanical properties and apatite-forming ability due to the increased nucleation sites provided, their high diffusion rates, reduced sintering time or temperature, and high mechanical properties. The aim of the present work was the evaluation of the effect of different ratios of Ethanol/TEOS and total amount of the inserted ammonia to the particle size, morphology and bioactive, hemolytic and antibacterial behavior of nanoparticles in the quaternary system SiO2-CaO-MgO-CuO. Different ratios of Ethanol/TEOS and ammonia amount affected the size and morphology of bioactive nanopowders. The optimum materials were synthesized with the highest ethanol/TEOS ratio and ammonia amount as verified by the enhanced apatite-forming ability and antibacterial and non-hemolytic properties.

Set-style yoghurts made from goat milk bases fortified with whey protein concentrates.

This research paper addresses the hypothesis that the fortification of goat milk base with whey protein concentrate (WPC) could affect both the textural and the biofunctional properties of set-style yoghurt. The effect of fortification of goat milk base with two different WPCs on thermophilic bacteria counts, proteolysis, physical and biofunctional properties of set-style yoghurts was studied at specific sampling points throughout a 4-week storage period. Fortification and storage did not influence thermophilic counts. Physical properties were affected significantly (P < 0.05) by the composition of the protein and the mineral fraction of the WPC but not by the storage. ACE-inhibitory activity was moderate in accordance to low lactobacilli counts and lack of proteolysis. DPPH-radical scavenging activity, Fe2+-chelating activity and superoxide scavenging activity were high. At 28 d an anti-inflammatory effect was observed, which was not affected by WPC addition.

In vitro evaluation of the shear bond strength and bioactivity of a bioceramic cement for bonding monolithic zirconia.

STATEMENT OF PROBLEM: Adhesive cementation is the most common bonding strategy for zirconia restorations. Although cementation with a bioactive luting agent has been proposed as an alternative, how the bond strength compares is unclear. PURPOSE: The purpose of this in vitro study was to evaluate shear bond strength after cementing a monolithic zirconia ceramic to human dentin with a bioceramic cement, compare it with a traditional cement, and evaluate its bioactive properties. MATERIAL AND METHODS: A total of 120 dentin specimens and 120 yttria-stabilized tetragonal zirconia polycrystal (Y-TZP) (BruxZir) cylindrical specimens were used. Zirconia and dentin specimens were randomly divided into 8 study groups (n=15) based on 2 luting cement types (a bioceramic cement or glass ionomer cement as control), 2 airborne-particle abrasion protocols (50 µm or 110 µm), and 2 water storage durations (24 hours or 30 days). After the shear bond strength test using a universal machine at a crosshead speed of 1 mm/min, fracture patterns were evaluated under a stereomicroscope and a scanning electron microscope. Strength values were statistically analyzed with a 3-factor ANOVA model (α=.05). Bioactivity was evaluated in simulated body fluid (SBF). RESULTS: The control glass ionomer cement achieved significantly greater shear bond strength compared with the tested bioceramic cement. Mean bond strength values ranged from 2.52 MPa to 5.23 MPa for the bioceramic cement tested and from 4.20 MPa to 6.61 MPa for the control cement. The duration of water storage played a significant role in the bond strength, with groups stored for 30 days reaching higher bond strength values, whereas the particle size of airborne-particle abrasion did not have a significant effect. Failure types were primarily mixed. No apatite formation was recorded on the surface of the specimens even after 30 days of immersion in SBF. CONCLUSIONS: The evaluated cement did not develop apatite in SBF, and its bond strength values were below the control glass ionomer cement.

Leucocyte expression of genes implicated in the plasminogen activation cascade is modulated by yoghurt peptides.

The urokinase-plasminogen activator (u-PA), its receptor (u-PAR) and the inhibitors of u-PA (PAI-1 and PAI-2) provide a multi-molecular system in leucocytes that exerts pleiotropic functions influencing the development of inflammatory and immune responses. The objective of the present study was to examine the ability of water soluble extracts (WSE) obtained from traditional Greek yoghurt made from bovine or ovine milk to modulate the expression of u-PA, u-PAR, PAI-1 and PAI-2 in ovine monocytes and neutrophils. WSE were obtained from 8 commercial traditional type Greek yoghurts made from ovine or bovine milk. WSE upregulated the expression of all 4 u-PA related genes in monocytes but the upregulation was much higher in the PAI-1 (10-fold) than in u-PA and u-PAR (3-4 fold) thus, shifting the system towards inhibition. In line with this observation, WSE reduced total and membrane-bound u-PA activity in monocytes. In neutrophils, WSE caused small (50-60%) but significant (P < 0·05) reductions in expression of u-PAR and PAI-2 but had no effect on expression of u-PA, PAI-1 and on total cell-associated and membrane-bound u-PA activity. WSE from yoghurts made from bovine or ovine milk were essentially equally effective in affecting the u-PA system except for the u-PAR gene in ovine neutrophils that was affected (reduced) by the ovine and not the bovine WSE. In conclusion, peptides present in WSE modulated the expression of u-PA related genes but the effect was much more prominent in monocytes than in neutrophils.

Antioxidant Activity of Sweet Whey Derived from Bovine, Ovine and Caprine Milk Obtained from Various Small-Scale Cheese Plants in Greece before and after In Vitro Simulated Gastrointestinal Digestion.

Whey-derived peptides have been associated with different biological properties, but most peptides are usually further hydrolyzed during the digestive process. In the present study, the antioxidant capacity of 48 samples of sweet whey (SW) derived from cheeses obtained from small-scale cheese plants made with bovine, ovine, caprine or a mixture of ovine/caprine milk was assessed using both cell-free and cell-based assays. SW digestates (SW-Ds) and a fraction (<3 kDa; SW-D-P3) thereof were obtained after in vitro digestion and subsequent ultrafiltration. Antioxidant properties using four different assays were evaluated before and after digestion. Our data showed higher values (p < 0.05) for ORAC, ABTS, FRAP and P-FRAP after in vitro digestion (SW-Ds and SW-D-P3) when compared with the corresponding values before digestion. In the non-digested SW, ORAC values were higher (p < 0.05) for the bovine SW compared with all the other samples. In contrast, the ABTS assay indicated a higher antioxidant activity for the ovine SW both before digestion and for SW-D-P3 compared with the bovine SW. The fraction SW-D-P3 of the ovine SW, using HT29 cells and H2O2 as an oxidizing agent, increased (p < 0.05) the cellular antioxidant activity. Furthermore, the same fraction of the ovine/caprine mixed SW increased, through the NF-κB pathway, the expression of SOD1 and CAT, genes implicated in the oxidative response in macrophage-like THP-1 cells. These findings indicate that SW, and particularly bovine and ovine SW, could be a candidate source for physical antioxidants in human and animal nutrition.

Effect of Milk Origin and Seasonality of Yogurt Acid Whey on Antioxidant Activity before and after In Vitro Gastrointestinal Digestion.

Yogurt acid whey (YAW) is a by-product of Greek strained yogurt production. The disposal of YAW constitutes an environmental problem, and given the increasing demand of Greek yogurt worldwide, its handling is a challenge. However, whey-derived peptides, resulting from microbial fermentation as well as those resulting from further hydrolysis during the digestion process, have been linked to enhanced biological activities. In this study, the antioxidant capacity of 33 samples of YAW obtained from Greek dairy companies of bovine, ovine or caprine origin was investigated using both cell-free and cell-based assays. The YAW samples, their in vitro digestion products (YAW-Ds) and a fraction of the digests (less than 3 kDa; YAW-D-P3) were assessed using four biochemical assays, namely ORAC, ABTS, FRAP and P-FRAP. Our data revealed a higher antioxidant capacity for digested samples compared with undigested samples, with all four methods. ORAC values after in vitro digestion were higher for the ovine samples compared to their bovine (YAW-D and YAW-D-P3) and caprine (YAW-D-P3) counterparts. Furthermore, the YAW-D-P3 fraction derived from samples collected in the summer months exhibited higher ORAC values when compared to the respective fraction from the winter months' samples. The cellular antioxidant activity of ovine YAW-D-P3 was improved in H2O2-treated HT29 cells compared to the control H2O2-treated cells. However, YAW-D-P3 could not trigger either the pathways involving the transcription factors NF-κB or NFE2L2 or the gene expression of SOD1, CAT and HMOX1 in LPS-challenged THP-1-derived macrophages. These results suggest that YAW, and particularly YAW from ovine origin, could be used as a natural source for its antioxidant potential in human and animal nutrition.

The Effects of Yoghurt Acid Whey Marination on Quality Parameters of Pork and Chicken Meat.

Large amounts of yoghurt acid whey (YAW) are annually generated as a result of Greek-style yoghurt production, which poses a great hazard to the environment. In terms of sustainability, YAW application in the meat industry appears as a great alternative since meat marination with natural solutions is a practice that continuously gains ground due to its positive effects on meat sensory attributes. The aim of the present study was to determine the quality characteristics and oxidative status of pork and chicken meat after their marination in yoghurt acid whey. Forty samples per meat type were randomly assigned into five groups: CON, without YAW marination; YAW1 and YAW3, in which meat was marinated at 4 °C and a pH of 4.5 for 15 and 10 h, respectively; or YAW2 and YAW4, in which meat was handled as in the YAW1 and YAW3 group, respectively, while hesperidin at 2 g/L was also incorporated into the marinade. As shown, meat shear force values were decreased in pork but not in chicken meat samples. Meat pH values were also generally decreased, while lightness was increased in raw but not in cooked meat samples as an effect of marination. Moreover, meat oxidative stability was improved to a greater extent in chicken than in pork meat. In order to find the ideal marination period for pork meat, we further immersed it into YAW for 5 h. However, this treatment affected neither meat tenderness and the other quality properties nor meat oxidation rates. In general, hesperidin addition did not have an additional or side effect on the quality traits of pork and chicken meat. As it can be concluded, pork meat marination in YAW for 10-15 h improves tenderness, but 5 h of marination does not. On the other hand, the tenderness of chicken meat was not affected, but its oxidative stability was greatly improved after remaining in the YAW marinade for 10-15 h.

Neuroimaging and Electroencephalographic Correlation in Patients with Transient Global Amnesia: Clinical Case Series.

Objective. To determine if there is any correlation between the electroencephalographic and neuroimaging findings in patients with Transient Global Amnesia (TGA). Methods: We retrospectively reviewed files of the First Department of Neurology of AHEPA University Hospital, including patients with a clinical diagnosis of TGA. Only patients who had the characteristic high signal in the temporal lobes in the DWI MRI and those who underwent electroencephalographic recording (EEG) were selected. Results: Out of 28 patients, 8 were selected. We found that 6 out of 8 patients (75%) who had imaging findings in DWI, in at least one medial temporal lobe, also had had intermittent slow theta waves on the electroencephalographic recording. Of these 6 patients, 3 (50%) had bilateral EEG findings, 2 patients (33,3%) only had findings on the left hemisphere and 1 (17%) had on the right hemisphere. 3 out of 6 patients (50%) had electroencephalographic dominance on the left, while 2 out of the 6 (33%) had on the right. In 2 patients with imaging findings in DWI no anomalies were demonstrated on EEG. In 3 out of 8 patients, both MRI and EEG findings correlated on the same side, while 1 patient had opposite findings, depending on which hemisphere the EEG anomalies dominated. Conclusions: There is no absolute matching between the DWI MRI and EEG findings in patients with the clinical diagnosis of TGA. However, there is some degree of correlation, when we focus on the focal dominance of the EEG anomalies, although not statistically significant.

Dietary Inclusion Level Effects of Yoghurt Acid Whey Powder on Performance, Digestibility of Nutrients and Meat Quality of Broilers.

In recent years, the increasing demand for Greek strained yoghurt produced massive amounts of acid whey, which constitutes a major environmental pollutant. Whether yoghurt acid whey can be included in poultry diets is not known. The purpose of this study was to evaluate the effects of four dietary levels of yoghurt acid whey powder (YAWP) on the growth performance, nutrient digestibility, meat quality traits and oxidative stability. A total of 300 male 1-day-old Ross 308 broilers were assigned into four groups with five replicates of 15 broilers each: control-fed basal diet with no YAWP addition (WO) or basal diet supplemented with YAWP at 25 g/kg of diet (WA), 50 g/kg of diet (WB), or 100 g/kg of diet (WC). At the starter period, body weight and body weight gain were reduced after WB and WC treatments compared to the WO treatment. Breast meat oxidative stability was improved during refrigerated storage for 1 and 3 d in all YAWP treatments compared to control, while the WA treatment showed an improved oxidative stability after 6 and 9 d. The results suggest that YAWP inclusion at 25 g/kg of diet did not impair performance and extended the meat shelf life by reducing lipid oxidation rates.

Soya protein hydrolysates modify the expression of various pro-inflammatory genes induced by fatty acids in ovine phagocytes.

The objective of the present study was to test the hypothesis that fatty acids are the circulating mediators acting in a pro-inflammatory manner towards activated circulating ovine monocyte/macrophages and neutrophils. Furthermore, whether soya protein hydrolysates (SPH) inhibit the fatty acid-induced increase in the production of pro-inflammatory responses by ovine phagocytes was tested in vitro. All the fatty acids tested (myristic, palmitic, palmitoleic, stearic and oleic) increased (P<0·01; C18>C16>C14) membrane-bound urokinase plasminogen activator (u-PA) and u-PA free binding sites in cell membranes of activated ovine blood monocytes/macrophages, but only the C18 fatty acids (stearic, oleic) were effective towards blood neutrophils. The C18 fatty acids up-regulated (P<0·05) the gene expression of u-PA, u-PA receptor, intercellular adhesion molecule 1 and inducible NO synthase (in monocytes) but not that of cyclo-oxygenase-2, integrin α X and plasminogen activator inhibitor types 1 and 2 by ovine phagocytes. SPH blocked completely or partially all C18 fatty acid-induced changes in the expression of various pro-inflammatory genes. In conclusion, fatty acids selectively 'activate' ovine phagocytes, suggesting that these cells 'sense' metabolic signals derived from adipocytes. Soya protein peptides inhibit all changes in gene expression induced by fatty acids in ovine phagocytes in vitro. This constitutes a novel mechanism of action.

COVID-19 mRNA Vaccination Mimicking Heart Attack in a Healthy 56-Year-Old Physician.

We report our experience regarding a 56-year-old physician who developed severe symptoms mimicking a heart attack a few days after receiving the second dose of the new mRNA vaccine of Pfizer-BioNTech for COVID-19 protection. The patient is a healthy individual with no comorbidities and a normal clinical and laboratory profile. Five days after receiving the second dose on his left shoulder, he manifested sudden, severe pain on the whole left arm which lasted for about one hour, gradually intensifying and migrating to the chest and presenting as severe angina or heart attack. All work-up, however, was negative, with no evidence of ischemic heart attack or myocarditis. Severe acute symptoms lasted for about 20 h and completely resolved after 36 h. Although myocarditis as a potential side effect of the vaccine with associated heart pain has been identified, chest pain mimicking heart attack with otherwise normal workup has not been reported. Physicians must consider this likely rare and self-resolving symptom in order to increase awareness and prevent themselves and their patients from increased anxiety and unnecessary laboratory investigations.

Morphofunctional examination of the carpal bones of pygmy hippopotamus from Ayia Napa, Cyprus.

This study provides a complete morphological description and functional analysis of the carpal bones of the endemic pygmy hippopotamus Phanourios minor, derived from the Upper Pleistocene site of Ayia Napa. From this deposit, numerous skeletal remains of this fossil hippo have been collected, making the locality one of the richest in Cyprus. The carpal bones were compared with those of extant hippopotamuses, to determine the changes that occurred in the fossil hippo. Examination of the elements showed that Phanourios presented some important features that were common among the endemic fossil ungulates of the Mediterranean islands. The carpal bones display a proximal-distal compression due to shortening of the distal part of the leg, due to the new ecological island conditions. However, they are robust with rough areas for strong muscular and ligament insertions, providing stability to the carpal joints, and low speed movement to the animal. The great flexor capabilities, and the limitation in ulnar deviation of the carpus, indicate that P. minor had increased agility in the sagittal plane and restricted transverse movements, while the suggestion of a more unguligrade stance for the species is ambiguous. Thus, the endemic Cypriot hippos developed specialized locomotion, suitable for walking on the rugged terrain of Cyprus, which seems to be different from that of its extant counterparts.

Functional morphology of the hindlimb of fossilized pygmy hippopotamus from Ayia Napa (Cyprus).

We studied the functional morphology of the postcranial skeleton of the endemic hippopotamus Phanourios minor, derived from the Upper Pleistocene site of Ayia Napa. The deposit, which consists of a hard limestone substrate on which the species moved, has yielded a great abundance of hippopotamus material, making the Ayia Napa locality one of the most important paleontological sites in Cyprus. The immigration of the large-sized mainland Hippopotamus to Cyprus led to the emergence of a new insular species with its main characteristic being the extremely reduced body size. In this study, all the hindlimb elements of the Cypriot hippo are described in detail and compared with those of the modern species, with the extant Hippopotamus amphibius being considered similar to the possible ancestor of P. minor. In some cases, the morphological comparison is reinforced using bones of other extinct insular and mainland hippos. Additionally, we provided a functional analysis of the hindlimb joints, suggesting specific locomotor habits for the species. The anatomical examination reveals that the elements in P. minor are robust with marked muscular insertion areas resembling those found in Hippopotamus. However, there are also similarities with Choeropsis liberiensis in certain morphofunctional traits. P. minor adapted to slow but powerful locomotion with remarkable stabilization, particularly in the zeugopodium and the autopodium. The knee was less mobile in the craniocaudal direction compared with that in recent hippos, while the abduction-adduction movements of the thigh were advanced. The pes presented good mobility in the sagittal plane and limitation in transversal movements. Thus, P. minor displayed modifications to its limbs, influenced by the mountainous island environment and the body size reduction, resulting in specialized locomotion, which was different from that of extant hippopotamuses.

Effects of Spaghetti Differing in Soluble Fiber and Protein Content on Glycemic Responses in Humans: A Randomized Clinical Trial in Healthy Subjects.

This randomized, single blind, cross-over study investigated the glycemic responses to three spaghetti No 7 types differing in dietary protein and soluble fiber content. Fourteen clinically and metabolically healthy, fasting individuals (25 ± 1 years; ten women; BMI 23 ± 1 kg/m2) received isoglucidic test meals (50 g available carbohydrate) and 50 g glucose reference, in random order. GI was calculated using the FAO/WHO method. Capillary blood glucose and salivary insulin samples were collected at 0, 15, 30, 45, 60, and 120 min. Subjective appetite ratings (hunger, fullness, and desire to eat) were assessed by visual analogue scales (VAS, 100 mm) at baseline and 120 min. All three spaghetti types (regular, whole wheat, and high soluble fiber-low carbohydrates) provided low GI values (33, 38, and 41, respectively, on glucose scale) and lower peak glucose values compared to glucose or white bread. No differences were observed between spaghetti No 7 types for fasting glucose, fasting and post-test-meal insulin concentrations, blood pressure (systolic and diastolic), and subjective appetite. Conclusions: all spaghetti No 7 types, regardless of soluble fiber and/or protein content, attenuated postprandial glycemic response, which may offer advantages to glycemic control.

The Effects of Replacing Soybean Meal with Rapeseed Meal, Cottonseed Cake, and Fava Beans on the Milk Yield and Quality Traits in Milking Ewes.

The replacement of soybean meal (SBM) from intensively reared dairy sheep diets has emerged as a significant challenge for sustainable production. However, the effects of this replacement on milk production have not been sufficiently elucidated. The objective of this study was to prospectively assess the effects of replacing SBM with a mixture of alternative protein sources on the milk yield (MY) and the milk quality traits (MQT) in intensively reared dairy sheep. A total of 112 multiparous, purebred milking ewes of the Chios and Frizarta breeds, from two intensive dairy sheep farms, were involved in the study, postweaning, and were assigned to either the control (CR) or the experimental ration (ER) group. In the ER, 3/4 of the SBM was replaced by a mixture of rapeseed meal, cottonseed cake, and fava beans, producing a ration of a similar nutritional value. MY, MQT, and body condition scores were recorded for each individual ewe monthly for a period of 4 months during lactation. The experimental ration was associated with beneficial effects on daily and 100-day fat yields and on the electrical conductivity of milk as an improved udder health status indicator, with no adverse effects on any of the rest of the studied milk production traits.