Precision mapping of an in vivo N-glycoproteome reveals rigid topological and sequence constraints.

N-linked glycosylation is a biologically important protein modification, but only a small fraction of modification sites have been mapped. We developed a "filter aided sample preparation" (FASP)-based method in which glycopeptides are enriched by binding to lectins on the top of a filter and mapped 6367 N-glycosylation sites on 2352 proteins in four mouse tissues and blood plasma using high-accuracy mass spectrometry. We found 74% of known mouse N-glycosites and discovered an additional 5753 sites on a diverse range of proteins. Sites almost always have the N-!P-[S|T]-!P (where !P is not proline) and rarely the N-X-C motif or nonconsensus sequences. Combining the FASP approach with analysis of subcellular glycosite localization reveals that the sites always orient toward the extracellular space or toward the lumen of ER, Golgi, lysosome, or peroxisome. The N-glycoproteome contains a plethora of modification sites on factors important in development, organ-specific functions, and disease.

Novel Gluconobacter oxydans strains selected from Kombucha with potential postbiotic activity.

Gastric and colorectal cancer are among the most frequently diagnosed malignancies of the gastrointestinal tract. Searching for methods of therapy that complements treatment or has a preventive effect is desirable. Bacterial metabolites safe for human health, which have postbiotic effect, are of interest recently. The study aimed to preliminary assessment of the safety, antimicrobial, and anti-cancer activity of cell-free metabolites of Gluconobacter oxydans strains isolated from Kombucha beverages as an example of the potential postbiotic activity of acetic acid bacteria (AAB). The study material consisted of five AAB strains of Kombucha origin and three human cell lines (gastric adenoma-AGS, colorectal adenoma-HT-29, and healthy cells derived from the endothelium of the human umbilical vein-HUVEC). Results of the study confirms the health safety and functional properties of selected AAB strains, including their potential postbiotic properties. The best potential anticancer activity of the AAB cell-free supernatants was demonstrated against AGS gastric adenoma cells. The conducted research proves the postbiotic potential of selected acetic acid bacteria, especially the KNS30 strain. KEY POINTS: •The beneficial and application properties of acetic acid bacteria are poorly studied. •Gluconobacter oxydans from Kombucha show a postbiotic activity. •The best anticancer activity of the G. oxydans showed against gastric adenoma.

The Use of Various Types of Waste Paper for the Removal of Anionic and Cationic Dyes from Aqueous Solutions.

This study examined the possibility of using various types of waste paper-used newsprint (NP), used lightweight coated paper (LWC), used office paper (OP), and used corrugated cardboard (CC)-for the removal of anionic dyes, Acid Red 18 (AR18) and Acid Yellow 23 (AY23), and cationic dyes, Basic Violet 10 (BV10) and Basic Red 46 (BR46), from aqueous solutions. The scope of this research included the characterization of sorbents (FTIR, SEM, BET surface area, porosity, pHPZC, effectiveness of water coloration), determination of pH effect on the effectiveness of dye sorption, sorption kinetics (pseudo-first-order model, second-order model, intraparticular diffusion model), and the maximum sorption capacity (Langmuir models and Freundlich model) of the tested sorbents. The use of waste paper materials as sorbents was found to not pose any severe risk of aquatic environment contamination. AR18, AY23, and BV10 sorption intensities were the highest at pH 2, and that of RB46 at pH 6. The waste paper sorbents proved particularly effective in removing cationic dyes, like in the case of, e.g., NP, which had a sorption capacity that reached 38.87 mg/g and 90.82 mg/g towards BV10 and BR46, respectively, and were comparable with that of selected activated carbons (literature data).

Acceptance of illness, quality of life and nutritional status of patients after lower limb amputation due to diabetes mellitus.

INTRODUCTION: Diabetes mellitus is one of the most common chronic diseases with a high number of sufferers worldwide. Diabetic neuropathy and diabetic angiopathy lead to serious infectious complications which are very difficult to combat and may finally lead to the amputation of a lower limb. The aim of the study was to evaluate the quality of life, the level of acceptance of the illness and the nutritional status of patients after lower limb amputation due to diabetes. METHODS: Ninety-nine patients (23 men and 76 women) were enrolled into the study. They had all undergone lower limb amputation due to diabetic foot and were treated in the Department of General and Oncological Surgery. The following questionnaires were used: the WHO Quality of Life-BREF (WHOQOL-BREF), the Mini Nutritional Assessment (MNA), the Acceptance of Illness Scale (AIS) and the anonymous specific socio-demographic characteristics questionnaire. RESULTS: The diabetes-related amputees were revealed to have a higher QoL within the social domain (mean score 64.48), an intermediate QoL - within the environmental domain (mean score 63.04) and the mental domain (mean score 59.61), and a lower QoL - within physical (somatic) domain (mean score 54.69). There was no statistical correlation between genders or between all the domains of QoL (P > 0.05). The mean MNA score was 22.66, which means that patients were at risk of malnutrition. There were statistical differences between women and men as regards nutritional status (P = 0.034). The mean AIS score was 27.65 (27.09 women and 29.48 men), which means that the respondents accept their disease. There was no statistical correlation between gender and the acceptance of the disease (P = 0.288). There was not statistical correlation between age and QoL (P > 0.05). There were statistical differences between age and nutritional status (P < 0.05), and between age and acceptance of the illness (P = 0.044). CONCLUSIONS: The better the quality of life was in all the domains, the better the level of acceptance of illness was. The less malnourished the patient was, the better their quality of life was in all the domains.

The Use of Unique, Environmental Lactic Acid Bacteria Strains in the Traditional Production of Organic Cheeses from Unpasteurized Cow's Milk.

The aim of this study was to use local LAB cultures for the production of organic acid-rennet cheeses from unpasteurized cow's milk. Under industrial conditions, three types of cheese were produced, i.e., traditionally with acid whey (AW), with starter culture L. brevis B1, or with starter culture L. plantarum Os2. Strains were previously isolated from traditional Polish cheeses. Chemical composition, physico-chemical, microbiological, and sensory studies during 2 months of storage were carried out. As a result of this research, it was found that the basic composition was typical for semi-hard, partially skimmed cheeses. Mainly saturated fatty acids were detected. The cheeses were rich in omega-3, -6, and -9 fatty acids and conjugated linoleic acid (CLA), and were characterized by good lipid quality indices (LQI). All of the cheeses were characterized by a high number of lactic acid bacteria, with Enterobacteriaceae, yeast, molds, and staphylococci contaminants, which is typical microbiota for unpasteurized milk products. Water activity, pH, and total acidity were typical. A lower oxidation-reduction potential (ORP) of cheeses with the addition of strains and stability of the products during storage were observed. The B1 and Os2 cheeses were lighter, less yellow, had a more intense milk and creamy aroma, were softer, moister, and more elastic than AW cheese. The research results indicate the possibility of using environmental LAB strains in the production of high-quality acid-rennet cheeses, but special attention should be paid to the production process due to the microbiological quality of the cheeses.

Human microbiome and homeostasis: insights into the key role of prebiotics, probiotics, and symbiotics.

The interest in the study of the gut microbiome has grown exponentially. Indeed, its impact on health and disease has been increasingly reported, and the importance of keeping gut microbiome homeostasis clearly highlighted. However, and despite many advances, there are still some gaps, as well as the real discernment on the contribution of some species falls far short of what is needed. Anyway, it is already more than a solid fact of its importance in maintaining health and preventing disease, as well as in the treatment of some pathologies. In this sense, and given the existence of some ambiguous opinions, the present review aims to discuss the importance of gut microbiome in homeostasis maintenance, and even the role of probiotics, prebiotics, and symbiotics in both health promotion and disease prevention.

Effects of Fructose and Oligofructose Addition on Milk Fermentation Using Novel Lactobacillus Cultures to Obtain High-Quality Yogurt-like Products.

The incorporation of prebiotics in fermented milk products is one of the best ways to promote health benefits while improving their sensory characteristics at the same time. The aim of this study was to evaluate the effects of the addition of fructose and oligofructose (1% and 2%) on the physicochemical, rheological, sensory, and microbiological quality attributes of fermented milk products inoculated with indigenous probiotic starter cultures of Lactobacillus isolated from Polish traditional fermented foods. The samples were evaluated during 35 days of refrigerated storage. The oligofructose and fructose caused increases in the populations of bacteria in comparison to the control fermented milk products without the addition of saccharides. The degrees of acidification in different fermented milk samples, as well as their viscosity, firmness, syneresis, and color attributes, changed during storage. The highest overall sensory quality levels were observed for the samples supplemented with L. brevis B1 and oligofructose. This study is the first attempt to compare the influences of different sugar sources on the physicochemical, rheological, sensory, and microbiological quality attributes of fermented milk products.

Cluster Analysis Classification of Honey from Two Different Climatic Zones Based on Selected Physicochemical and of Microbiological Parameters.

The geographical origin of honey affects its composition, which is of key importance for the health-promoting properties and safety of the product. European regulations clearly define the physicochemical requirements for honey that determine the microbiological quality. On the other hand, legislation abolishes microbiological criteria. In the study 40 honey samples originating from two different climatic zones were analyzed. The water content, pH, water activity analysis and the microbiological quality of honey samples have been tested using the reference plate method (total viable count, yeast and molds, lactic acid bacteria, Bacillus spp.). The cluster classification showed that total viable count of bacteria could be used as a measure alternative to the count of Bacillus spp. and 70% of honeys from the tropical climate zone had different microbiological quality than honeys from the temperate climate zone but still under the level 3.0 log cfu/g. The study has revealed that geographical origin of honey may significantly affect the quality and safety of honey. It was considered that water content can be the most informative and handy marker of the microbiological quality of honeys. Analysis of lactic acid bacteria showed temperate climate zone honeys as a source of beneficial bacteria in the diet.

Effects of Lacticaseibacillus rhamnosus LOCK900 on Development of Volatile Compounds and Sensory Quality of Dry Fermented Sausages.

Traditional dry fermented meat products are highly appreciated by consumers. A probiotic starter culture increases their attractiveness through sensory qualities and a potential health-promoting effect. The ability to scale the laboratory solution to industrial conditions is an additional scientific and practical value of a new way of using probiotics in the meat industry. The aim was to evaluate the influence of the probiotic starter culture Lacticaseibacillus rhamnosus LOCK900 on the development of volatile organic compounds and the sensory quality of dry fermented pork sausages during fermentation and refrigeration storage. The microbiological and sensory characteristic (QDA method) and volatile compound (gas chromatography coupled with mass spectrometry: GC-MS) were evaluated. The number of LOCK900 cells during 12 weeks of storage remained above 6 log CFU g-1, making this product a functional food. The addition of probiotic LOCK900 increased the levels of acidic volatile compounds, aldehydes, and esters, which, combined with the additives and spices used, had a positive effect on the sensory properties of ripening sausages. The sausages with LOCK900 were characterised by positive sensory features, and their overall quality remained high during storage and did not differ from that of the control sausages.

Mapping N-glycosylation sites across seven evolutionarily distant species reveals a divergent substrate proteome despite a common core machinery.

N-linked glycosylation is an important posttranslational modification in all eukaryotes, but little is known about the N-glycoproteomes in nonmammalian systems. Here, we measure N-glycoproteomes of the major model organisms Arabidopsis thaliana, Schizosaccharomyces pombe, Saccharomyces cerevisiae, Caenorhabditis elegans, Drosophila melanogaster, and Danio rerio, representatively spanning the eukaryotic domain of life. The number of detected N-glycosylation sites varied between 425 in fission yeast, 516 in budding yeast, 1,794 in worm, 2,186 in plant, 2,229 in fly, and 2,254 in zebrafish. We find that all eukaryotic N-glycoproteomes have invariant characteristics including sequence recognition patterns, structural constraints, and subcellular localization. However, a surprisingly large percentage of the N-glycoproteome evolved after the phylogenetic divergences between plants, fungi, nematodes, insects, and vertebrates. Many N-glycosylated proteins coevolved with the rise of extracellular processes that are specific within corresponding phylogenetic groups and essential for organismal development, body growth, and organ formation.

Probiotics: Versatile Bioactive Components in Promoting Human Health.

The positive impact of probiotic strains on human health has become more evident than ever before. Often delivered through food, dietary products, supplements, and drugs, different legislations for safety and efficacy issues have been prepared. Furthermore, regulatory agencies have addressed various approaches toward these products, whether they authorize claims mentioning a disease's diagnosis, prevention, or treatment. Due to the diversity of bacteria and yeast strains, strict approaches have been designed to assess for side effects and post-market surveillance. One of the most essential delivery systems of probiotics is within food, due to the great beneficial health effects of this system compared to pharmaceutical products and also due to the increasing importance of food and nutrition. Modern lifestyle or various diseases lead to an imbalance of the intestinal flora. Nonetheless, as the amount of probiotic use needs accurate calculations, different factors should also be taken into consideration. One of the novelties of this review is the presentation of the beneficial effects of the administration of probiotics as a potential adjuvant therapy in COVID-19. Thus, this paper provides an integrative overview of different aspects of probiotics, from human health care applications to safety, quality, and control.

The Effect of Molecular Weight on the Antibacterial Activity of N,N,N-Trimethyl Chitosan (TMC).

N,N,N-trimethyl chitosan (TMC) with 93% degree of trimethylation was synthesized. TMC and the chitosan starting material were subjected to acidic hydrolysis to produce 49 different samples with a reduced average molecular weight (Mw) ranging from 2 to 144 kDa. This was done to allow the investigation of the relationship between antibacterial activity and Mw over a wide Mw range. NMR investigation showed that hydrolysis did not affect the degree of trimethylation (DSTRI) or the structure of the polymer backbone. The activity of TMC against Staphylococcus aureus (S. aureus) increased sharply with Mw until a certain Mw value (critical Mw for high activity, CMW) was reached. After the CMW, the activity was not affected by a further increase in the Mw. A similar pattern of activity was observed for chitosan. The CMW was determined to be 20 kDa for TMC and 50 kDa for chitosan.

The use of different dialysis membranes in therapy of patients with multiple myeloma.

Free light chains accumulation is the reason of kidney injury in patients with multiple myeloma. The removal of free light chains can improve patients prognosis and survival, and in some cases allows for dialysotherapy discontinuation. Unfortunately, conventional dialysis is not effective enough in terms of free light chains removal. New high cut-off (HCO) techniques remove free light chains more effectively than conventional dialysis. In some cases, this technique may turn out better than hemodiafiltration. However, there are some differences between specific techniques in the removal of kappa and lambda light chains. Lambda light chains are better removed by polymethyl methacrylate membranes with a change of filter during dialysis. Kappa light chains are thoroughly removed by polymethyl methacrylate membranes and HCO (35,000 Da) polysulfone membranes. Unfortunately, it is very difficult to differentiate between the effect of HCO dialysis therapy and concomitant chemotherapy because some of the data is not fully conclusive. Using the proper technique for an individual patient may give optimally effective treatment results.

Design of New-Generation Usable Forms of Topical Haemostatic Agents Containing Chitosan.

Designing usable forms of topical haemostatic agents is the most important activity during the design process, resulting in strengthened functional properties of the final medical devices. This study aimed to propose indications for a research programme based on risk management supporting the development of two usable forms of a topical haemostatic agent: chitosan/alginate lyophilized foam and chitosan/alginate impregnated gauze. Both of the usable forms of the topical haemostatic agent, being the main part of the modified combat gauze, were fabricated using the chitosan/alginate complex. Risk analysis is helpful in developing an appropriate research programme, significantly reducing the risk to an acceptable level.

Bacteriocins from lactic acid bacteria as an alternative to antibiotics.

Bacteriocins are ribosomally synthesized, proteinaceous substances that inhibit the growth of closely related species through numerous mechanisms. The classification system used in this review divided bacteriocins into four sub-groups based on their size. Currently, there is extensive research focused on bacteriocins and their usage as a food preservative. The increasing incidence of multidrug resistant bacterial pathogens is one of the most pressing medical problems in recent years. Recently, the potential clinical application of LAB (Lactic Acid Bacteria) bacteriocin has been the subject of investigations by many scientists. Bacteriocins can be considered in a sense as antibiotic, although they differ from conventional antibiotics in numerous aspects. The gene-encoded nature of bacteriocins makes them easily amenable through bioengineering to either increase their activity or specify target microorganism. Owing to this feature of bacteriocins, antibiotic therapy would become less damaging to the natural gut microflora, which is a common drawback of conventional antibiotic use. Bacteriocins from lactic acid bacteria represent one of the most studied microbial defense systems and the idea of subjecting them to bioengineering to either increase antimicrobial activity or further specify their target microorganism is now a rapidly expanding field. This review aimed to present bacteriocins as a possible alternative to conventional antibiotics basic on latest scientific data.

Cognitive disorders in children's hydrocephalus.

BACKGROUND AND OBJECTIVE: Hydrocephalus is defined as an increase of volume of cerebrospinal fluid in the ventricular system of the brain. It develops as a result of cerebrospinal fluid flow disorder due to dysfunctions of absorption or, less frequently, as a result of the increase of its production. Hydrocephalus may lead to various cognitive dysfunctions in children. MATERIALS AND METHODS: In order to determine cognitive functioning in children with hydrocephalus, the authors reviewed available literature while investigating this subject. RESULTS: The profile of cognitive disorders in children with hydrocephalus may include a wide spectrum of dysfunctions and the process of neuropsychological assessment may be very demanding. The most frequently described cognitive disorders within children's hydrocephalus include attention, executive, memory, visual, spatial or linguistic dysfunctions, as well as behavioral problems.

In vitro screening of selected probiotic properties of Lactobacillus strains isolated from traditional fermented cabbage and cucumber.

Most important during probiotic selection are gastric acid and bile tolerance, the adhesion to the luminal epithelium to colonize the lower gastrointestinal tract of a human and safety for human consumption. The aim of this study was to evaluate the selected probiotic in vitro properties of Lactobacillus spp. Strains isolated from traditional fermented food. A total 38 strains were isolated from the pickled samples and 14 were identified as Lactobacillus spp. The survival of almost all strains after incubation at pH 2.5 did not change markedly, and remained at above 90 % (10(9) CFU/mL). The strains also exhibited a high survival rate at pH 3.5 (>90 %), whereas pH 1.5 all were died. Just four strains could survive 90 min. at pH 1.5 (<39 %). The incubation with 0.2 % bile salt solution resulted in a survival rates of 81-94 % after 24 h, whereas after incubation in 2 and 4 % bile salt solution it was 59-94 %. All tested strains showed very good and good resistance to 0.4 % phenol addition, however only Lb. johnsonii K4 was able to multiply. The hydrophobic nature of the cell surface of the tested strains was moderated recording hydrophobicity of Lb. johnsonii K4 and Lb. rhamnosus K3 above 60 %. Safety evaluation excluded four of tested strains as candidate probiotics, according to antibiotic resistance patterns and certain metabolic activities. On the basis on the results 10 of the selected Lactobacillus strains are safe and can survive under gastrointestinal conditions, which requires them to future in vitro and in vivo probiotic studies.

The Interplay between Gut Microbiota and Cognitive Functioning in the Healthy Aging Population: A Systematic Review.

BACKGROUND: The gut microbiota in healthy older individuals typically show a decrease in beneficial bacteria like Bifidobacterium and Lactobacillus, alongside an increase in pro-inflammatory microbes such as Enterobacteriaceae and Clostridia. These changes contrast with younger and middle-aged individuals and appear to correlate with cognitive status. Although there is extensive research on gut microbiota and cognitive functions in cognitively impaired elderly individuals, its impact on cognitively healthy elderly populations has not been extensively studied. METHOD: A comprehensive literature search was conducted across PubMed, EBSCO, Web of Science, and Scopus databases to identify studies exploring the relationship between gut microbiota composition and cognitive functioning in healthy older adults. During the literature screening process, each record was initially assessed by its title, abstract, and keywords to exclude articles that did not align with the scope of this review. Three authors independently screened and retrieved the records. The inclusion criteria included: (1) publication in peer-reviewed journals; (2) studies involving neurologically, cognitively, and medically healthy populations; (3) participants identified as older adults, defined for this review as individuals aged 45 years and older due to the limited number of records; (4) analysis of gut microbiota; and (5) assessment of cognitive function. Subsequently, full texts were analyzed to determine eligibility. The exclusion criteria encompassed: (1) incorrect publication type; (2) inappropriate sample population; (3) unsuitable study design; (4) absence of one or more inclusion criteria; and (5) studies based on animal research. A risk of bias assessment was performed for each included study using the Joanna Briggs Institute (JBI) checklist, ensuring all selected studies met established quality standards. RESULTS: A total of 6 eligible research articles from a possible 1752 published until March 2024 were identified and included. We categorized the included studies into two groups based on their focus: the taxonomic composition of gut microbiota and the alpha diversity, which is the variety of organisms within a sample. Additionally, two methods were identified for assessing cognition: neuropsychological tests and physiological measurements, notably electroencephalography (EEG). The studies show varying results regarding the abundance of specific bacterial taxa and their cognitive associations. Notably, the relationship between certain bacteria and cognition may vary when analyzed at different taxonomic levels, such as phylum versus family. CONCLUSIONS: Changes in gut microbiota composition in the elderly, even without a cognitive impairment diagnosis, could potentially serve as early biological markers for Alzheimer's disease or other dementias before mild cognitive impairment appears.

The Role of Autochthonous Levilactobacillus brevis B1 Starter Culture in Improving the Technological and Nutritional Quality of Cow's Milk Acid-Rennet Cheeses-Industrial Model Study.

In the study, an attempt was made to develop an innovative technology for cheese manufacturing. It was hypothesized that selected autochthonous lactic acid bacteria as a starter culture are more suitable for the production of acid-rennet cheeses of good technological and sensory quality. The study aimed to assess the possibility of using the strain Levilactobacillus brevis B1 (L. brevis B1) as a starter culture to produce acid-rennet cheeses using raw cow's milk. Two variants of cheese were manufactured. The control variant (R) was coagulated with microbial rennet and buttermilk, and the other variant (B1) was inoculated with rennet and L. brevis B1 starter culture. The effect of the addition of these autochthonous lactic acid bacteria on selected physicochemical characteristics, durability, the composition of fatty acids, cholesterol, Iipid Quality Indices, and microbiological and sensory quality of acid-rennet cheeses was determined during a 3-month period of storage. The dominant fatty acids observed in the tested cheeses were saturated fatty acids (SFA) (68.43-69.70%) and monounsaturated fatty acids (MUFA) (25.85-26.55%). Significantly higher polyunsaturated fatty acid (PUFA) content during storage was observed for B1 cheeses. The B1 cheeses were characterized by lower cholesterol content compared to cheese R and showed better indexes, including the Index of atherogenicity, Index of thrombogenicity, DFA, OFA, H/H, and HPI indexes, than the R cheese. No effect of the tested L. brevis B1 on sensory quality was observed in relation to the control cheeses during 3 months of storage. The results of the research indicate the possibility of using the L. brevis B1 strain for the production of high-quality, potentially probiotic acid-rennet cheeses.

Extensive quantitative remodeling of the proteome between normal colon tissue and adenocarcinoma.

We report a proteomic analysis of microdissected material from formalin-fixed and paraffin-embedded colorectal cancer, quantifying > 7500 proteins between patient matched normal mucosa, primary carcinoma, and nodal metastases. Expression levels of 1808 proteins changed significantly between normal and cancer tissues, a much larger fraction than that reported in transcript-based studies. Tumor cells exhibit extensive alterations in the cell-surface and nuclear proteomes. Functionally similar changes in the proteome were observed comparing rapidly growing and differentiated CaCo-2 cells. In contrast, there was minimal proteomic remodeling between primary cancer and metastases, suggesting that no drastic proteome changes are necessary for the tumor to propagate in a different tissue context. Additionally, we introduce a new way to determine protein copy numbers per cell without protein standards. Copy numbers estimated in enterocytes and cancer cells are in good agreement with CaCo-2 and HeLa cells and with the literature data. Our proteomic data set furthermore allows mapping quantitative changes of functional protein classes, enabling novel insights into the biology of colon cancer.