Correction: Gastric coagulation and postprandial amino acid absorption of milk is affected by mineral composition: a randomized crossover trial.

Correction for 'Gastric coagulation and postprandial amino acid absorption of milk is affected by mineral composition: a randomized crossover trial' by Elise J. M. van Eijnatten et al., Food Funct., 2024, 15, 3098-3107, https://doi.org/10.1039/D3FO04063A.

Gastric coagulation and postprandial amino acid absorption of milk is affected by mineral composition: a randomized crossover trial.

Background: In vitro studies suggest that casein coagulation of milk is influenced by its mineral composition, and may therefore affect the dynamics of protein digestion, gastric emptying and appearance of amino acids (AA) in the blood, but this remains to be confirmed in vivo. Objective: This study aimed to compare gastrointestinal digestion between two milks with the same total calcium content but different casein mineralization (CM). Design: Fifteen males (age 30.9 ± 13.8 years, BMI 22.5 ± 2.2 kg m-2) participated in this randomized cross-over study with two treatments. Participants underwent gastric magnetic resonance imaging (MRI) scans at the baseline and every 10 min up to 90 min after consumption of 600 ml milk with low or high CM. Blood samples were taken at the baseline and up to 5 hours postprandially. Primary outcomes were postprandial plasma AA concentrations and gastric emptying rate. Secondary outcomes were postprandial glucose and insulin levels, gastric coagulation as estimated by image texture metrics, and appetite ratings. Results: Gastric content volume over time was similar for both treatments. However, gastric content image analysis suggested that the liquid fraction emptied quicker in the high CM milk, while the coagulum emptied slower. Relative to high CM, low CM showed earlier appearance of AAs that are more dominant in casein, such as proline (MD 4.18 µmol L-1, 95% CI [2.38-5.98], p < 0.001), while there was no difference in appearance of AAs that are more dominant in whey protein, such as leucine. The image texture metrics homogeneity and busyness differed significantly between treatments (MD 0.007, 95% CI [0.001, 0.012], p = 0.022; MD 0.005, 95% CI [0.001, 0.010], p = 0.012) likely because of a reduced coagulation in the low CM milk. Conclusions: Mineral composition of milk can influence postprandial serum AA kinetics, likely due to differences in coagulation dynamics. The clinical trial registry number is NL8959 (https://clinicaltrials.gov).

[Closing the vaccine gap: pregnant women in the Netherlands will be offered a flu jab]. / Meer grip op griep bij zwangere vrouwen.

Influenza immunisation during pregnancy is recommended by the WHO since 2010 and implemented in the majority of the Western countries. Finally, the Dutch government follows this recommendation and will be offering vaccination to pregnant women starting upcoming influenza season. Remarkably, this new guideline mainly focuses on the benefits for infants. In this article we want to make healthcare workers aware of the fact that the maternal influenza vaccination has substantial benefits for both mothers and their infants. Vaccination will reduce the risk of infection and influenza-associated hospitalizations. Potentially, the vaccination has positive effects on pregnancy outcomes like birth weight and premature delivery, but further research is needed. Offering the vaccination requires a solid campaign, in which all involved healthcare workers should advocate for vaccination. In addition, the vaccination should not only be available for pregnant women above 22 weeks of gestational age, but throughout the entire pregnancy if necessary.

Platelets express adaptor proteins of the extrinsic apoptosis pathway and can activate caspase-8.

BACKGROUND: Apoptotic pathways in platelets are important for their survival and function. Platelet apoptosis may be involved in the pathogenesis of immune thrombocytopenia (ITP), an autoimmune-mediated disease. In contrast to the intrinsic apoptosis pathway, not much is known about the extrinsic pathway mechanisms in platelets. OBJECTIVES: To investigate the expression of proteins involved in the extrinsic apoptosis pathway, including the death receptors, adaptor and regulator proteins in human platelets. To determine a possible trigger of the extrinsic apoptosis pathway in platelets. METHODS: To investigate the expression of key markers of the extrinsic pathway we used targeted immunofluorescence and flow cytometry assays. To study their expression and interaction we performed Western blotting and co-immunoprecipitation. Treated platelets with different apoptosis triggers were subjected to flow cytometry. RESULTS: We could identify the protein expression of the pro-apoptotic proteins TRADD (Tumor Necrosis Factor Receptor type 1- Associated DEATH Domain protein), TRAF2/5, (TNF Associated Factor) and DEDAF (Death Effector Domain- Associated Factor), FADD (Fas-Associated protein with death domain) as well as the anti-apoptotic proteins DJ-1 (Deglycase 1) and c-FLIP in human platelets. ABT-737 treatment induced a disruption in the co-localization of DJ-1 with FADD. Platelets treated with ABT-737 showed an activation in caspase-3 and -8. The exposure to TNF (Tumor Necrosis Factor), FasL (Fas ligand), and TWEAK or to plasma derived from ITP patients, did not lead to changes in caspase-3 and -8 activation in platelets. CONCLUSIONS: Human platelets express some proteins of the extrinsic apoptosis pathway which can be modulated only by ABT-737 treatment. However so far, no other apoptosis trigger or interaction with an external receptor have been yet identified.

Increased levels of IL-10 and IL-1Ra counterbalance the proinflammatory cytokine pattern in acute pediatric immune thrombocytopenia.

Immune thrombocytopenia (ITP) is an autoimmune disease which leads to accelerated platelet clearance. We investigated the plasma cytokine, chemokine and growth factor signatures and their clinical significance in pediatric ITP patients during acute, chronic and follow-up stages as well as the effects of intravenous immunoglobulin (IVIg) treatment, by using the Multiplex technology. In acute ITP before and/or after IVIg treatment we found significantly increased plasma levels of the pro- (tumour necrosis factor-α (TNF-α), interleukin IL-15) and anti- (IL-1 receptor antagonist (Ra), IL-10 and the growth factor interferon γ-induced protein (IP-10)) inflammatory cytokines, compared to healthy controls. Except for IL1-Ra, these cytokines decreased to normal levels in chronic patients. In contrast, growth-regulated α protein (GRO) and soluble CD40 ligand (sCD40L), known as platelet-derived molecules, were found to be significantly decreased in acute and increased in chronic ITP patients compared to healthy controls. GRO levels positively correlated with the platelet counts in the follow-up and chronic cohort. Monocyte counts showed a significant positive correlation only with IP-10 levels in acute ITP after IVIg treatment and follow-up patients. Expression levels of mRNAs for macrophage inflammatory protein MIP1-ß, IL-1Ra and GRO determined in peripheral blood mononuclear cells (PBMCs) were significantly reduced in both acute and chronic ITP compared to controls. Our findings suggest that the different clinical presentation of acute and chronic pediatric ITP and to a lesser extent the IVIg treatment effects are characterized overall by a counterbalanced cytokine, chemokine and growth factor pattern response that might exert a pathogenic role in this disease.

Simplifying the diagnosis of inherited platelet disorders? The new tools do not make it any easier.

The molecular causes of many inherited platelet disorders are being unraveled. Next-generation sequencing facilitates diagnosis in 30% to 50% of patients. However, interpretation of genetic variants is challenging and requires careful evaluation in the context of a patient's phenotype. Before detailed testing is initiated, the treating physician and patient should establish an understanding of why testing is being performed and discuss potential consequences, especially before testing for variants in genes associated with an increased risk for hematologic malignancies.

An international external quality assessment for laboratory diagnosis of heparin-induced thrombocytopenia.

Essentials A pilot study for External Quality Assessment for testing of HIT is described. The qualitative accordance for the PF4/heparin IgG test was 97.6%. The qualitative accordance for functional HIT tests was considerably lower. External Quality Assessment for functional HIT tests is required. SUMMARY: Objective Heparin-induced thrombocytopenia (HIT) is a potentially life-threatening complication of heparin exposure. Diagnosis is most reliable using a combination of an enzyme immunoassay (EIA) that detects antibodies against platelet factor 4 (PF4)/heparin complexes ("antigen" assay) and a "functional" assay that detects platelet-activating properties of the pathogenic HIT antibodies. No External Quality Assessment (EQA) is available for a combination of the tests. Here we report on the results of the first international EQA. Methods The pilot EQA was organized by the Department of Transfusion Medicine, Universitätsmedizin Greifswald, Germany. Six serum samples of patients, which were referred to Greifswald for HIT diagnosis, and one negative control sample were distributed to seven participants in Germany, Canada, and Singapore. Participants were asked to report the optical density (OD) values of their local EIA test for IgG-specific antibodies against the PF4/heparin complexes and the results for a functional assay (HIPA or SRA). Consensus was defined as a minimum 70% agreement, i.e., agreement among at least five of the seven participating laboratories. Results and conclusion Six out of seven participants reported results for EIA, with a high quantitative accordance (97.6%). For the functional assay, consensus was reached for all samples except the negative control, for which some participants reported nonspecific reactivity. All HIT-negative samples were correctly diagnosed by all participants; for HIT-positive samples, consensus of 70% was reached. Although the limited availability of sample material is an obstacle to overcome, an EQA combining both EIA and functional testing is feasible.

Diagnosis of hereditary platelet disorders in the era of next-generation sequencing: "primum non nocere".

Inherited platelet disorders can affect "only platelets", occur as a "syndromic phenotype" or be associated with "increased risk of hematological malignancies". Genetic testing is attractive for diagnosis of inherited platelet disorders. However, many physicians who refer patient blood for genetic testing are unaware of the association of certain inherited platelet disorders with other risks. Inherited platelet disorders associated with minor-moderate bleeding rarely cause patient distress. In contrast, identification of a mutation associated with an increased risk of leukemia may cause a major psychological disease burden, without offsetting the beneficial impact on management. Guidelines recommend postponing genetic testing "until the patient reaches adulthood or at least until the child is mature enough to participate in decision making". In our opinion, outside research, (genetic) testing in children with inherited platelet disorders should only be performed if it influences management. In adults, genes causing inherited platelet disorders associated with an increased risk of hematological malignancies should only be tested after obtaining explicit informed consent.

Hb G-Waimanalo [A1] [α64(E13)Asp→Asn; HBA1: c.193 G > A] with Decreased Oxygen Affinity.

A clinically asymptomatic 12-year-old girl showed microcytosis in routine examination. Cation exchange high performance liquid chromatography (HPLC), revealed two additional peaks eluting after Hb A and DNA sequencing uncovered a novel heterozygous mutation at codon 64 of the α1-globin gene. The hemoglobin (Hb) variant was annotated as Hb G-Waimanalo [A1]. Further analyses demonstrated a decreased oxygen affinity Hb compared to the normal Hb configuration.

Deep sequencing and proteomic analysis of the microRNA-induced silencing complex in human red blood cells.

During maturation, erythropoietic cells extrude their nuclei but retain their ability to respond to oxidant stress by tightly regulating protein translation. Several studies have reported microRNA-mediated regulation of translation during terminal stages of erythropoiesis, even after enucleation. In the present study, we performed a detailed examination of the endogenous microRNA machinery in human red blood cells using a combination of deep sequencing analysis of microRNAs and proteomic analysis of the microRNA-induced silencing complex. Among the 197 different microRNAs detected, miR-451a was the most abundant, representing more than 60% of all read sequences. In addition, miR-451a and its known target, 14-3-3ζ mRNA, were bound to the microRNA-induced silencing complex, implying their direct interaction in red blood cells. The proteomic characterization of endogenous Argonaute 2-associated microRNA-induced silencing complex revealed 26 cofactor candidates. Among these cofactors, we identified several RNA-binding proteins, as well as motor proteins and vesicular trafficking proteins. Our results demonstrate that red blood cells contain complex microRNA machinery, which might enable immature red blood cells to control protein translation independent of de novo nuclei information.

Antiviral strategies combining antiretroviral drugs with RNAi-mediated attack on HIV-1 and cellular co-factors.

To improve the care of HIV-1/AIDS patients there is a critical need to develop tools capable of blocking viral evolution and circumventing therapy-associated problems. An emerging solution is gene therapy either as a stand-alone approach or as an adjuvant to pharmacological drug regimens. Combinatorial RNAi by multiplexing antiviral RNAi inhibitors through vector-mediated delivery has recently shown significant superiority over conventional mono-therapies. Viral as well as cellular co-factor targets have been identified, but they are generally attacked separately. Here, we hypothesized that a mixture of shRNAs directed against highly conserved viral RNA sequences and the mRNAs of cellular components that are involved in HIV replication could restrict mutational escape by enhanced synergistic inhibition. We screened for potent silencer cocktails blending inhibitors acting scattered along the viral replication cycle. The results show enhanced and extended suppression of viral replication for some combinations. To further explore the power of combinatorial approaches, we tested the influence of RNAi-mediated knockdown on the activity of conventional antiretroviral drugs (fusion, RT, integrase and protease inhibitors). We compared the fold-change in IC50 (FCIC50) of these drugs in cell lines stably expressing anti-HIV and anti-host shRNAs and measured increased values that are up by several logs for some combinations. We show that high levels of additivity and synergy can be obtained by combining gene therapy with conventional drugs. These results support the idea to validate the therapeutic potential of this anti-HIV approach in appropriate in vivo models.

Toward a durable treatment of HIV-1 infection using RNA interference.

RNA interference (RNAi) is a cellular mechanism that mediates sequence-specific gene silencing at the posttranscriptional level. RNAi can be used as an antiviral approach against human pathogens. An attractive target for RNAi therapeutics is the human immunodeficiency virus type 1 (HIV-1), and the first clinical trial using a lentiviral gene therapy was initiated in early 2008. In this chapter, we focus on some basic principles of such an RNAi-based gene therapy against HIV-1. This includes the subjects of target site selection within the viral RNA genome, the phenomenon of viral escape, and therapeutic strategies to prevent viral escape. The latter antiescape strategies include diverse combinatorial RNAi approaches that are all directed against the HIV-1 RNA genome. As an alternative strategy, we also discuss the possibilities and restrictions of targeting cellular cofactors that are essential for virus replication, but less important for cell physiology.

Directed adenovirus evolution using engineered mutator viral polymerases.

Adenoviruses (Ads) are the most frequently used viruses for oncolytic and gene therapy purposes. Most Ad-based vectors have been generated through rational design. Although this led to significant vector improvements, it is often hampered by an insufficient understanding of Ad's intricate functions and interactions. Here, to evade this issue, we adopted a novel, mutator Ad polymerase-based, 'accelerated-evolution' approach that can serve as general method to generate or optimize adenoviral vectors. First, we site specifically substituted Ad polymerase residues located in either the nucleotide binding pocket or the exonuclease domain. This yielded several polymerase mutants that, while fully supportive of viral replication, increased Ad's intrinsic mutation rate. Mutator activities of these mutants were revealed by performing deep sequencing on pools of replicated viruses. The strongest identified mutators carried replacements of residues implicated in ssDNA binding at the exonuclease active site. Next, we exploited these mutators to generate the genetic diversity required for directed Ad evolution. Using this new forward genetics approach, we isolated viral mutants with improved cytolytic activity. These mutants revealed a common mutation in a splice acceptor site preceding the gene for the adenovirus death protein (ADP). Accordingly, the isolated viruses showed high and untimely expression of ADP, correlating with a severe deregulation of E3 transcript splicing.

Long-term inhibition of HIV-1 replication with RNA interference against cellular co-factors.

In this study we tested whether HIV-1 replication could be inhibited by stable RNAi-mediated knockdown of cellular co-factors. Cell lines capable of expressing shRNAs against 30 candidate co-factors implicated at different steps of the viral replication cycle were generated and analyzed for effects on cell viability and inhibition of HIV-1 replication. For half of these candidate co-factors we obtained knockdown cell lines that are less susceptible to virus replication. For three co-factors (ALIX, ATG16 and TRBP) the cell lines were resistant to HIV-1 replication for up to 2 months. With these cells we could test the hypothesis that HIV-1 is not able to escape from RNAi-mediated suppression of cellular co-factors, which was indeed not detected.

[Study of calcification of the mitral annulus. Importance of echocardiographic analysis]. / Estudio de la calcificación del anillo mitral. Importancia del análisis ecocardiográfico.

Mitral annulus calcification is a common finding in old people. In order to know the association of mitral annulus calcification with other pathologic conditions, 25 patients were studied by echocardiography, 20 females and 5 males. No significant differences _ere observed with respect to sex. The average age was 67 +/- 15 years. Mitral annulus calcification was associated with aortic sclerosis in 100% of the cases, to arterial hypertension in 19 (76%), to obstructive pulmonary disease in 8 (32%), to diabetes mellitus in 5 (20%), and to peripheral arterial insufficiency in 5 (20%). Mitral regurgitation was found in 14 cases, atrial fibrillation in 2 (8%). Complete A-V block was not observed, but there was enlargement of the left ventricle in 23 (92%) and of the left atrium in 17 (68%). The chest X-ray showed mitral annulus calcification in 5 (20%) with a sensitivity of 25% and a specificity of 100%. The M-mode echocardiogram showed LA-Ao ratio of 1.4 +/- 0.3, the mitral valve D-E excursion was reduced 11.9 +/- 3.1 mm. and also the E-F slope 28.6 mm/seg +/- 16.7 with appearance of mitral stenosis, but the two-dimensional study demonstrated that this was false. In all patients the left ventricle was dilated and fractional shortening was diminished. Echocardiography not only is a good diagnostic method for mitral annulus calcification, it also allow us to evaluate the hemodynamic consequences of this pathology which occurs in older patients and is often associated with other common illnesses of advanced age. In our study 56% of mitral annulus calcification cases were associated with mitral regurgitation.

[Natural history of rheumatic carditis. A follow-up of more than 20 years]. / Historia natural de la carditis reumática. Seguimiento por más de veinte años.

In this retrospective study of 43 patients of the National Institute of Cardiology of Mexico, 20 to 35 years after the first attack of rheumatic carditis shows that the prognosis of the heart valve disease is directly influenced by the number of attacks of rheumatic fever. In fact, when patients had only one rheumatic attack the secuelae was mild mitral regurgitation (19%), without hemodynamic significance, with less proportion of mitral stenosis (15%), or aortic regurgitation (7.6%), less plurivalvular lesions (16%), or required heart surgery (15%). In the other hand, when the patients suffered three rheumatic attacks had more proportion of mitral stenosis (33%), aortic regurgitation (41%), pluryvalvular lesions (38%) and required more heart (50%). We conclude that prophylactic treatment is important in patients with chronic rheumatic heart disease.

[Hypertensive retinopathy in patients with aortic coarctation]. / Retinopatía hipertensiva en pacientes con coartación aórtica.

In order to assess the incidence and the determinants of hypertensive vascular changes in the retina of patients with coarctation of the aorta, fifty (37 male and 13 female) were analyzed. The mean age of the group was 18.7 +/- 10.3 years. No one had evidence of nephropathy. Different degrees of hypertensive retinopathy (Puig-Solanes classification) were observed in 54%. Of the patients, no one had papilledema. Retinal vascular damage was not related to either age or sex. The group of patients with retinal vascular lesions had a mean level of systolic arterial pressure higher than the group with normal retinas. Multivariant statistical analysis of the results permitted the identification of systolic arterial pressure higher than 150 mm Hg, age greater than 15 years and cardiomegaly as the three variables more frequently associated with retinal vascular lesions. Surgical correction of the aortic malformation resulted in normalization of both systolic and diastolic arterial pressures in 98% of the total number of patients as determined one year postoperatively. It is concluded that, in coarctation of the aorta, vascular damage of the retina appears to have an incidence that is similar to that observed in patients with other forms of systemic arterial hypertension. The development of these retinal lesions in patients with coarctation of the aorta would seem to be determined by the severity and duration of the hypertensive process.