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This study describes the effect of a human milk protein on the activation of peripheral blood lymphocytes. A folding variant of [alpha]-lactalbumin from human milk has been isolated and characterized. The active complex precipitated with the casein fraction at pH 4.6 and was purified from casein by a combination of anion exchange and gel chromatography. The fraction showed N-terminal and mass spectrometric identity with human milk [alpha]-lactalbumin, but monomeric, native [alpha]-lactalbumin had no effect on blood lymphocytes. The active fraction was previously shown to induce apoptotic cell death in immature undifferentiated cells but spared normal cells. This human milk fraction was identified as multimeric [alpha]-lactalbumin (MAL). Our continued studies with MAL have focused on the hypothesis that milk components may have a modulating effect on the development of the immune system in breast-fed infants very early in life. We compared the effect of short-term exposure to MAL, monomeric [alpha]-lactalbumin and human milk casein on mononuclear cell activation by flow cytometry using CD69 expression as the measure of early activation. In contrast to monomeric [alpha]-lactalbumin and casein, MAL proved to have a strong activating effect on lymphocytes leading to a fivefold increase in expression. Analysis of the lymphocyte activation response brings us one step closer to proving our hypothesis.

Background: On average, people in the United States visit a doctor 4 times a year, and many of them have chronic illnesses. Because of the increased use of technology, people frequently rely on the internet to access health information and statistics. People use health care information to make better-educated decisions for themselves and others. Health care dashboards should provide pertinent and easily understood data, such as information on timely cancer screenings, so the public can make better-informed decisions. In order to enhance health outcomes, effective dashboards should provide precise data in an accessible and easily digestible manner., Objective: This study identifies the top 15 attributes of a health care dashboard. The objective of this research is to enhance health care dashboards to benefit the public by making better health care information available for more informed decisions by the public and to improve population-level health care outcomes., Methods: The authors conducted a survey of health care dashboards with 218 individuals identifying the best practices to consider when creating a public health care dashboard. The data collection was conducted from June 2023 to August 2023. The analyses performed were descriptive statistics, frequencies, and a comparison to a prior study., Results: From May 2023 to June 2023, we collected 3259 responses in multiple different states around the United States from 218 people aged 18 years or older. The features ranking in descending order of importance are as follows: (1) easy navigation, (2) historical data, (3) simplicity of design, (4) high usability, (5) use of clear descriptions, (6) consistency of data, (7) use of diverse chart types, (8) compliance with the Americans with Disabilities Act, (9) incorporated user feedback, (10) mobile compatibility, (11) comparison data with other entities, (12) storytelling, (13) predictive analytics with artificial intelligence, (14) adjustable thresholds, and (15) charts with tabulated data., Conclusions: Future studies can extend the research to other types of dashboards such as bioinformatics, financial, and managerial dashboards as well as confirm these top 15 best practices for medical dashboards with further evidentiary support. The medical informatics community may benefit from standardization to improve efficiency and effectiveness as dashboards can communicate vital information to patients worldwide on critically prominent issues. Furthermore, health care professionals should use these best practices to help increase population health care outcomes by informing health care consumers to make better decisions with better data., (©Melina Malkani, Eesha Madan, Dillon Malkani, Arav Madan, Neel Singh, Tara Bamji, Harman Sabharwal. Originally published in the Online Journal of Public Health Informatics (https://ojphi.jmir.org/), 20.11.2024.)

Purpose: The present randomized-controlled trial was conducted to assess the effect of Kangen water and reverse osmosis (RO) water on dental plaque, salivary pH and salivary Streptococcus mutans count., Materials and Methods: This randomized control trial was conducted for 14 days on 24 randomly selected participants from the pool of undergraduate dental students. Participants were randomly divided into two groups of 12 each: the Kangen water (pH 9) group and the RO water group. Participants in each group were asked to drink allocated water for 7 days. Dental plaque, salivary pH and microbial colony-forming units (CFUs) were assessed after 7 and 14 days., Results: Intragroup comparison showed that all three outcomes showed a significant improvement in the Kangen water group after 14 days, whereas no difference was seen in the RO water group. Intergroup comparison showed a significant difference in plaque score and CFU among the two groups after 7 and 14 days, whereas pH between the two groups did not show a significant difference., Conclusions: Regular drinking of alkaline Kangen water with pH 9 was found to be effective in reducing plaque and salivary Streptococcus mutans count when compared to RO water.

Background: In randomised controlled trials (RCTs), the application of a test of significance to compare the baseline differences between the intervention groups is a common practice, though it has been condemned by many researchers., Objective: This study aimed to assess the proportion of RCTs on human participants comparing the baseline differences between intervention groups using the test of significance in nine dental specialty journals published in India and to estimate the proportion of studies reporting baseline demographic and clinical characteristics in a table., Materials and Methods: RCTs published in nine dental journals published by dental specialty associations of India were screened. A literature search was limited to the time duration of five years from 2017 to 2021., Results: The authors analysed 326 RCTs. Of 326 RCTs published, 237 RCTs (72.7%) did not report the baseline demographic and clinical characteristics table. Tests of significance were applied to compare baseline differences between the intervention arms in 148 (45.4%) RCTs published., Conclusions: Although criticised by the Consolidated Standards of Reporting Trials (CONSORT) statement, the majority of the trials published in dental specialty journals failed to avoid comparison of baseline differences with significance test and failed to report baseline characteristic table., Competing Interests: None

Background: Mature aggressive B-cell lymphomas are heterogenous malignancies that make up more than half of all diagnosed non-Hodgkin lymphoma in children and adolescents. The overall survival rate increased over the last decades to 80%-90% due to fine tuning of polychemotherapy. However, new therapeutic implications are needed to further increase the overall survival. Current clinical trials analyze the therapeutic effect of rituximab in pediatric patients, while the mechanism of action in vivo is still not fully understood., Methods: Effector molecules important for tumor defense were analyzed before and at day 5 after rituximab treatment via flow cytometry. Serum rituximab levels were measured with an ELISA., Results: We evaluated patient parameters that may affect treatment response in relation to rituximab administration and serum rituximab levels. We indeed found a reduction of Fcγ receptor (FcγR) II levels after rituximab treatment in monocyte subtypes, whereas FcγRI expression was significantly increased. Serum levels of proinflammatory marker proteins S100A8/A9 and S100A12 significantly decreased after treatment to normal levels from an overall proinflammatory state before treatment. CD57, perforin, and granzyme B expression decreased after treatment, comprising a less cytolytic natural killer (NK) cell population., Conclusion: The highlighted effects of rituximab treatment on patient's immune response help in understanding the biology behind tumor defense mechanisms and effector function. After subsequent studies, these novel insights might be translated into patient care and could contribute to improve treatment of pediatric patients with mature aggressive B-cell lymphoma., (© 2021 The Authors. Pediatric Blood & Cancer published by Wiley Periodicals LLC.)

Bacterium-host interactions in the gut proceed via directly contacted epithelial cells, the host's immune system, and a plethora of bacterial factors. Here we characterized and compared exemplary cytokine and microRNA (miRNA) responses of human epithelial and THP-1 cells toward the prototype enteropathogenic Escherichia coli (EPEC) strain E2348/69 (O127:H6) and the probiotic strain Escherichia coli Nissle 1917 (EcN) (O6:K5:H1). Human T84 and THP-1 cells were used as cell culture-based model systems for epithelial and monocytic cells. Polarized T84 monolayers were infected apically or basolaterally. Bacterial challenges from the basolateral side resulted in more pronounced cytokine and miRNA responses than those observed for apical side infections. Interestingly, the probiotic EcN also caused a pronounced transcriptional increase of proinflammatory CXCL1 and interleukin-8 (IL-8) levels when human T84 epithelial cells were infected from the basolateral side. miR-146a, which is known to regulate adaptor molecules in Toll-like receptor (TLR)/NF-κB signaling, was found to be differentially regulated in THP-1 cells between probiotic and pathogenic bacteria. To assess the roles of flagella and flagellin, we employed several flagellin mutants of EcN. EcN flagellin mutants induced reduced IL-8 as well as CXCL1 responses in T84 cells, suggesting that flagellin is an inducer of this cytokine response. Following infection with an EPEC type 3 secretion system (T3SS) mutant, we observed increased IL-8 and CXCL1 transcription in T84 and THP-1 cells compared to that in wild-type EPEC. This study emphasizes the differential induction of miR-146a by pathogenic and probiotic E. coli strains in epithelial and immune cells as well as a loss of probiotic properties in EcN interacting with cells from the basolateral side., (Copyright © 2016, American Society for Microbiology. All Rights Reserved.)

Tightly controlled epithelial and endothelial barriers are a prerequisite for life as these barriers separate multicellular organisms from their environment and serve as first lines of defense. Barriers between neighboring epithelial cells are formed by multiple intercellular junctions including the 'apical junctional complex-AJC' with tight junctions (TJ), adherens junctions (AJ), and desmosomes. TJ consist of tetraspan transmembrane proteins like occludin, various claudins that directly control paracellular permeability, and the 'Junctional Adhesion Molecules' (JAMs). For establishing tight barriers TJ are essential but at the same time have to allow also selective permeability. For this, TJ need to be tightly regulated and controlled. This is organized by a variety of adaptor molecules, i.e., protein kinases, phosphatases and GTPases, which in turn are regulated and fine-tuned involving microRNAs (miRNAs). In this review we summarize available data on the role and targeting of miRNAs in the maintenance of epithelial and/or endothelial barriers.

Rev is an essential HIV-1 regulatory protein which binds to the Rev responsive element (RRE) present within the env gene of HIV-1 RNA genome. This binding facilitates the transport of the RNA to the cytoplasm, which in turn triggers the switch between viral latency and active viral replication. Essential components of this complex have been localized to a minimal arginine rich Rev peptide and stem IIB region of RRE. A synthetic peptide known as RSG-1.2 binds with high binding affinity and specificity to the RRE-IIB than the Rev peptide, however the thermodynamic basis of this specificity has not yet been addressed. The present study aims to probe the thermodynamic origin of this specificity of RSG-1.2 over Rev Peptide for RRE-IIB. The temperature dependent melting studies show that RSG-1.2 binding stabilizes the RRE structure significantly (ΔT(m) = 4.3°C), in contrast to Rev binding. Interestingly the thermodynamic signatures of the binding have also been found to be different for both the peptides. At pH 7.5, RSG-1.2 binds RRE-IIB with a K(a) = 16.2±0.6×10(7) M(-1) where enthalpic change ΔH = -13.9±0.1 kcal/mol is the main driving force with limited unfavorable contribution from entropic change TΔS = -2.8±0.1 kcal/mol. A large part of ΔH may be due to specific stacking between U72 and Arg15. In contrast binding of Rev (K(a) = 3.1±0.4×10(7) M(-1)) is driven mainly by entropy (ΔH = 0 kcal/mol and TΔS = 10.2±0.2 kcal/mol) which arises from major conformational changes in the RNA upon binding.

RNA-peptide interactions have been intensively studied at the structural level; however, in the absence of thermodynamic studies, the molecular forces that dictate the binding specificities and affinities remain elusive. Here we evaluate the thermodynamics (DeltaG, DeltaH, DeltaS) of HIV-1 TAR RNA hairpin and Tat peptide interaction as well as the hydration changes that accompany these interactions, through a series of calorimetric, spectroscopic, and osmotic stress studies. Tat peptide binding enhances the thermal stability of the TAR RNA hairpin; however, the thermal enhancement decreases with increasing Na(+) concentration. The equilibrium association constant (K(a)) is determined by fluorescence titrations and examined as a function of Na(+) concentration and temperature. The binding constant (K(a)) decreases with increasing Na(+) concentration. The binding free energy (DeltaG) is found to have a large nonpolyelectrolyte component with release of a single counterion upon binding. The ITC profiles showed two independent sites binding, indicating specific as well as nonspecific interactions. The enthalpy changes associated with both the binding sites are found to be more negative for the binding process at lower salt concentration of 20 mM Na(+). Our binding studies under osmotic stress conditions show that there is a release of 28 (+/-4) and 21 (+/-3) water molecules upon complex formation at 20 and 80 mM Na(+) concentration supporting the observed positive entropy contributions and accounting for discrepancies between DeltaH(cal) and DeltaH(vH). In aggregate, our results suggest that the hydrogen bonding of arginine to TAR RNA dictates the binding interaction.

Previous studies have shown that human serum containing anti-group A streptococcus carbohydrate (GAS CHO) antibodies were opsonic for different M protein-carrying serotypes. To investigate the role that anti-GAS CHO antibodies play in passive and active protection, mice were immunized subcutaneously or intranasally with GAS CHO conjugated to tetanus toxoid, and mortality and oral colonization were monitored after challenge with live GAS. Compared with control mice, immunized mice were significantly protected against systemic or nasal challenge with GAS. Furthermore, studies of serum samples and throat cultures from Mexican children revealed an inverse relationship between high serum titers of anti-GAS CHO antibodies and the presence of GAS in the throat. Anti-GAS CHO antibodies were also tested for cross-reactivity with human tissues and cytoskeletal proteins. No cross-reactivity was observed in either assay. The present study demonstrates that GAS CHO is both immunogenic and protective against GAS infections.

This study describes an alpha-lactalbumin folding variant from human milk with bactericidal activity against antibiotic-resistant and -susceptible strains of Streptococcus pneumoniae. The active complex precipitated with the casein fraction at pH 4.6 and was purified from casein by a combination of anion exchange and gel chromatography. Unlike other casein components, the active complex was retained on the ion-exchange matrix and eluted only with high salt. The eluted fraction showed N-terminal and mass spectrometric identity with human milk alpha-lactalbumin, but native alpha-lactalbumin had no bactericidal effect. Spectroscopic analysis demonstrated that the active form of the molecule was in a different folding state, with secondary structure identical to alpha-lactalbumin from human milk whey, but fluctuating tertiary structure. Native alpha-lactalbumin could be converted to the active bactericidal form by ion-exchange chromatography in the presence of a cofactor from human milk casein, characterized as a C18:1 fatty acid. Analysis of the antibacterial spectrum showed selectivity for streptococci; Gram-negative and other Gram-positive bacteria were resistant. The folding variant of alpha-lactalbumin is a new example of naturally occurring molecules with antimicrobial activity.

Background: The establishment of the nasopharyngeal flora was followed in Costa Rican children from birth to 1 year of age., Methods: Nasopharyngeal cultures were obtained at 1 (n = 413), 3 (n = 393), 6 (n = 376) and 12 months (n = 356) of age from children representative of the population in the Puriscal district. Weekly cultures were obtained from a subcohort of these children (n = 101). Mother-infant diads (n = 95) and preschool children (n = 208) attending day-care centers were also studied., Results: The estimated proportion of colonized children in the population differed markedly depending on the frequency of culture. Quarterly cultures showed a slow increase in carrier rates from 3.9% for Haemophilus influenzae, 3.1% for Streptococcus pneumoniae and 6.5% for Moraxella catarrhalis at 1 month of age to 10.1% carrying H. influenzae and 19.4% carrying S. pneumoniae by the end of the first year. By quarterly culture the proportion of children colonized at least once was 36% for S. pneumoniae, 26% for H. influenzae and 28% for M. catarrhalis. In contrast weekly sampling showed that 95 to 100% of the children were colonized at least once during the first year of life with H. influenzae, S. pneumoniae or M. catarrhalis. Nasopharyngeal carriage of H. influenzae, S. pneumoniae and M. catarrhalis was low in the mothers, and very few mother-infant pairs carried identical bacteria at the same time. In contrast carrier rates were high in the siblings attending day care (H. influenzae 27.9%, S. pneumoniae 39.4%, both organisms 26.6%). Infants with siblings had significantly higher bacterial carriage at all ages than infants without siblings., Conclusions: Quarterly nasopharyngeal cultures showed that Costa Rican infants acquire their nasopharyngeal flora at a rate comparable with that for infants in developed countries and that siblings are an important source of the bacteria. Weekly samplings showed that virtually all children were colonized at least once during the first year of life.

Haemophilus influenzae and Streptococcus pneumoniae are common causes of respiratory tract infections. H. influenzae attach to receptor epitopes in mucins and in epithelial cell membranes. Attachment is followed by an epithelial cell cytokine response. Secreted cytokines then initiate inflammation, upset the integrity of the mucosal barrier, and lead to disease. S. pneumoniae do not bind to mucins but attach to respiratory tract epithelial cells. Attachment is increased by viral infection of the epithelial cells. Unlike H. Influenzae, S. pneumoniae induce apoptosis in epithelial cells, thus disrupting the mucosal barrier. Attachment and persistence is counterbalanced by antiadhesive as well as bactericidal molecules in secretions such as human milk. These examples illustrate the balance between host defenses and microbial virulence as it has coevolved to maintain the health of the respiratory mucosa.

Melkersson-Rosenthal syndrome (MRS) is a rare condition with variable presentation. In some cases there is sequential development of clinical features. This report describes the syndrome in two Arab children who showed partial response to therapy with oral steroids. Since the natural course of the disease is unpredictable and there may be natural remission of symptoms, the efficacy of steroids is difficult to establish.

Eight oligosaccharides have been isolated from faeces of a blood group B, secretor, breast-fed infant and characterized by sugar and methylation analysis, f.a.b. mass spectrometry and 1H-n.m.r. spectroscopy. One of these oligosaccharides has not previously been reported and is a tri-L-fucosyl derivative of lacto-N-hexaose. The other compounds were identical to oligosaccharides found in human milk. Several of the reported compounds require the secretor dependent 2'-fucosyltransferase for their biosynthesis. Since the mother of this child was an O(H) non-secretor, an intestinal biosynthesis of at least some of these compounds is strongly indicated. No blood group B active oligosaccharides were detected which is in sharp contrast to the oligosaccharide excretion in faeces from a blood group A infant [Sabharwal et al., Mol. Immunol., 21 (1984) 1105-1112] in which all the major oligosaccharides isolated were blood group A active.

Nine neutral and five acidic oligosaccharides were isolated from feces of a preterm (30th postmenstrual week) blood group A nonsecretor infant fed on pooled breast milk. Structural analyses were carried out using sugar and methylation analyses, fast atom bombardment mass spectrometry, and 1H NMR. The acidic oligosaccharides are well-known components of human milk. The neutral oligosaccharides are characteristic of nonsecretor milk. Surprisingly, no secretor gene-dependent oligosaccharides were present in the feces. Another preterm (27th postmenstrual week) blood group A, secretor infant fed on pooled breast milk showed the same fecal oligosaccharide pattern as above during the first week after birth, despite being a secretor individual. Also notable was the absence of blood group A-active oligosaccharides in this sample. Another sample of feces collected 8 weeks later from the latter infant contained the expected blood group A-active oligosaccharides. Furthermore, free sialic acid was present at the cost of the sialyl oligosaccharides seen earlier. Thus, infants born prematurely do not show the same degree of development of oligosaccharide metabolism as their more mature counterparts.