Research priorities to reduce the impact of COVID-19 in low- and middle-income countries.

Background: The COVID-19 pandemic has caused disruptions to the functioning of societies and their health systems. Prior to the pandemic, health systems in low- and middle-income countries (LMIC) were particularly stretched and vulnerable. The International Society of Global Health (ISoGH) sought to systematically identify priorities for health research that would have the potential to reduce the impact of the COVID-19 pandemic in LMICs. Methods: The Child Health and Nutrition Research Initiative (CHNRI) method was used to identify COVID-19-related research priorities. All ISoGH members were invited to participate. Seventy-nine experts in clinical, translational, and population research contributed 192 research questions for consideration. Fifty-two experts then scored those questions based on five pre-defined criteria that were selected for this exercise: 1) feasibility and answerability; 2) potential for burden reduction; 3) potential for a paradigm shift; 4) potential for translation and implementation; and 5) impact on equity. Results: Among the top 10 research priorities, research questions related to vaccination were prominent: health care system access barriers to equitable uptake of COVID-19 vaccination (ranked 1st), determinants of vaccine hesitancy (4th), development and evaluation of effective interventions to decrease vaccine hesitancy (5th), and vaccination impacts on vulnerable population/s (6th). Health care delivery questions also ranked highly, including: effective strategies to manage COVID-19 globally and in LMICs (2nd) and integrating health care for COVID-19 with other essential health services in LMICs (3rd). Additionally, the assessment of COVID-19 patients' needs in rural areas of LMICs was ranked 7th, and studying the leading socioeconomic determinants and consequences of the COVID-19 pandemic in LMICs using multi-faceted approaches was ranked 8th. The remaining questions in the top 10 were: clarifying paediatric case-fatality rates (CFR) in LMICs and identifying effective strategies for community engagement against COVID-19 in different LMIC contexts. Interpretation: Health policy and systems research to inform COVID-19 vaccine uptake and equitable access to care are urgently needed, especially for rural, vulnerable, and/or marginalised populations. This research should occur in parallel with studies that will identify approaches to minimise vaccine hesitancy and effectively integrate care for COVID-19 with other essential health services in LMICs. ISoGH calls on the funders of health research in LMICs to consider the urgency and priority of this research during the COVID-19 pandemic and support studies that could make a positive difference for the populations of LMICs.

Synthesis and Evaluation of Imidazo[1,2-a]pyridine Analogues of the ZSTK474 Class of Phosphatidylinositol 3-Kinase Inhibitors.

Using a scaffold-hopping approach, imidazo[1,2-a]pyridine analogues of the ZSTK474 (benzimidazole) class of phosphatidylinositol 3-kinase (PI3K) inhibitors have been synthesized for biological evaluation. Compounds were prepared using a heteroaryl Heck reaction procedure, involving the palladium-catalysed coupling of 2-(difluoromethyl)imidazo[1,2-a]pyridines with chloro, iodo or trifluoromethanesulfonyloxy (trifloxy) substituted 1,3,5-triazines or pyrimidines, with the iodo intermediates being preferred in terms of higher yields and milder reaction conditions. The new compounds maintain the PI3K isoform selectivity of their benzimidazole analogues, but in general show less potency.

Long-Term In Vivo Oxygen Sensors for Peripheral Artery Disease Monitoring.

Tracking of tissue oxygenation around chronic foot wounds may help direct therapy decisions in patients with peripheral artery disease (PAD). Novel sensing technology to enable such monitoring was tested over 9 months in a Sinclair mini-pig model. No adverse events were observed over the entire study period. Systemic and acute hypoxia challenges were detected during each measurement period by the microsensors. The median time to locate the sensor signal was 13 s. Lumee Oxygen microsensors appear safe for long-term repeated oxygen measurements over 9 months.

Synthesis and biological evaluation of sulfonamide analogues of the phosphatidylinositol 3-kinase inhibitor ZSTK474.

Replacement of one of the morpholine groups of the phosphatidylinositol 3-kinase (PI3K) inhibitor ZSTK474 (1) with sulfonamide containing substituents produced a new class of active and potent PI3Kα inhibitors. Solubility issues prevented all but the 6-amino derivative 17 from being evaluated in vivo, but the clear activity of this compound demonstrated that this class of PI3K inhibitor shows great promise.

Tissue-Integrating Oxygen Sensors: Continuous Tracking of Tissue Hypoxia.

We describe a simple method of tracking oxygen in real-time with injectable, tissue-integrating microsensors. The sensors are small (500 µm × 500 µm × 5 mm), soft, flexible, tissue-like, biocompatible hydrogel s that have been shown to overcome the foreign body response for long-term sensing. The sensors are engineered to change luminescence in the presence of oxygen or other analytes and function for months to years in the body. A single injection followed by non-invasive monitoring with a hand-held or wearable Bluetooth optical reader enables intermittent or continuous measurements. Proof of concept for applications in high altitude, exercise physiology, vascular disease, stroke, tumors, and other disease states have been shown in mouse, rat and porcine models. Over 90 sensors have been studied to date in humans. These novel tissue-integrating sensors yield real-time insights in tissue oxygen fluctuations for research and clinical applications.

Biological characterization of SN32976, a selective inhibitor of PI3K and mTOR with preferential activity to PI3Kα, in comparison to established pan PI3K inhibitors.

Multiple therapeutic agents have been developed to target the phosphatidylinositol 3-kinase (PI3K) signaling pathway, which is frequently dysregulated in cancer promoting tumor growth and survival. These include pan PI3K inhibitors, which target class Ia PI3K isoforms and have largely shown limited single agent activity with narrow therapeutic windows in clinical trials. Here, we characterize SN32976, a novel pan PI3K inhibitor, for its biochemical potency against PI3K isoforms and mTOR, kinase selectivity, cellular activity, pharmacokinetics, pharmacodynamics and antitumor efficacy relative to five clinically-evaluated pan PI3K inhibitors: buparlisib, dactolisib, pictilisib, omipalisib and ZSTK474. SN32976 potently inhibited PI3K isoforms and mTOR, displaying preferential activity for PI3Kα and sparing of PI3Kδ relative to the other inhibitors, while showing less off-target activity than the clinical inhibitors in a panel of 442 kinases. The major metabolites of SN32976 were also potent PI3K inhibitors with similar selectivity for PI3Kα as the parent compound. SN32976 compared favorably with the clinically-evaluated PI3K inhibitors in cellular assays, inhibiting pAKT expression and cell proliferation at nM concentrations, and in animal models, inducing a greater extent and duration of pAKT inhibition in tumors than pictilisib, dactolisib and omipalisib at similarly tolerated dose levels and inhibiting tumor growth to a greater extent than dactolisib and ZSTK474 and with similar efficacy to pictilisib and omipalisib. These results suggest that SN32976 is a promising clinical candidate for cancer therapy with enhanced kinase selectivity and preferential inhibition of PI3Kα compared to first generation pan PI3K inhibitors, while retaining comparable anticancer activity.

Anti-histone H1 IgGs possess proliferative activity towards human T-leukaemia CEM cells.

The aim of this study was to characterize the proliferative activity of the anti-histone H1 IgGs towards human T-leukaemia CEM cells. MATERIALS AND METHODS: Anti-histone H1 IgGs were purified from blood serum of systemic lupus erythematosus patients by precipitation of serum proteins with 50% ammonium sulfate followed by a sequential affinity chromatography on Protein G-Sepharose and histone H1-Sepharose columns. To avoid contamination with other proteins, anti-histone H1 IgGs were subjected to strongly acidic pH 2.0 during gel filtration through HPLC column. The effects of the anti-histone H1 IgGs on cell viability and cell cycle were tested by MTS-assay and flow cytometry, correspondingly. The cross-reactivity of the anti-histone H1 antibodies towards heterogenetic and cellular antigens was evaluated by Western-blot analysis. RESULTS: It was found that incubation of CEM cells with the HPLC-purified anti-histone H1 IgGs resulted in significant stimulation of cell growth by 46% after 48 h of incubation. These IgGs possess an antigenic poly-specificity to positively charged heterogenetic antigens and different cellular antigens. FITC-labeled and biotinylated anti-histone H1 IgGs are internalized by CEM cells and preferentially accumulated in the cytoplasm. CONCLUSION: The anti-histone H1 IgGs are shown to internalize human T-leukemia CEM and stimulate their proliferation. These IgGs are polyspecific toward cellular antigens.

Desialylation of dying cells with catalytically active antibodies possessing sialidase activity facilitate their clearance by human macrophages.

Recently we reported the first known incidence of antibodies possessing catalytic sialidase activity (sialidase abzymes) in the serum of patients with multiple myeloma and systemic lupus erythematosus (SLE). These antibodies desialylate biomolecules, such as glycoproteins, gangliosides and red blood cells. Desialylation of dying cells was demonstrated to facilitate apoptotic cell clearance. In this study we assessed the possibility to facilitate dying cell clearance with the use of F(ab)2 fragments of sialidase abzymes. Two sources of sialidase abzymes were used: (i) those isolated from sera of patients with SLE after preliminary screening of a cohort of patients for sialidase activity; and (ii) by creating an induced sialidase abzyme through immunization of a rabbit with synthetic hapten consisting of a non-hydrolysable analogue of sialidase reaction conjugated with bovine serum albumin (BSA) or keyhole limpet haemocyanin (KLH). Antibodies were purified by ammonium sulphate precipitation, protein-G affinity chromatography and size exclusion-high performance liquid chromatography (HPLC-SEC). Effect of desialylation on efferocytosis was studied using human polymorphonuclear leucocytes (PMN), both viable and aged, as prey, and human monocyte-derived macrophages (MoMa). Treatment of apoptotic and viable prey with both disease-associated (purified from blood serum of SLE patients) and immunization-induced (obtained by immunization of rabbits) sialidase abzymes, its F(ab)2 fragment and bacterial neuraminidase (as positive control) have significantly enhanced the clearance of prey by macrophages. We conclude that sialidase abzyme can serve as a protective agent in autoimmune patients and that artificial abzymes may be of potential therapeutic value.

Oral zinc supplementation for the treatment of acute diarrhea in children: a systematic review and meta-analysis.

Evidence supporting the impact of therapeutic zinc supplementation on the duration and severity of diarrhea among children under five is largely derived from studies conducted in South Asia. China experiences a substantial portion of the global burden of diarrhea, but the impact of zinc treatment among children under five has not been well documented by previously published systematic reviews on the topic. We therefore conducted a systematic literature review, which included an exhaustive search of the Chinese literature, in an effort to update previously published estimates of the effect of therapeutic zinc. We conducted systematic literature searches in various databases, including the China National Knowledge Infrastructure (CNKI), and abstracted relevant data from studies meeting our inclusion and exclusion criteria. We used STATA 12.0 to pool select outcomes and to generate estimates of percentage difference and relative risk comparing outcomes between zinc and control groups. We identified 89 Chinese and 15 non-Chinese studies for the review, including studies in 10 countries from all WHO geographic regions, and analyzed a total of 18,822 diarrhea cases (9469 zinc and 9353 control). None of the included Chinese studies had previously been included in published pooled effect estimates. Chinese and non-Chinese studies reported the effect of therapeutic zinc supplementation on decreased episode duration, stool output, stool frequency, hospitalization duration and proportion of episodes lasting beyond three and seven days. Pooling Chinese and non-Chinese studies yielded an overall 26% (95% CI: 20%-32%) reduction in the estimated relative risk of diarrhea lasting beyond three days among zinc-treated children. Studies conducted in and outside China report reductions in morbidity as a result of oral therapeutic zinc supplementation for acute diarrhea among children under five years of age. The WHO recommendation for zinc treatment of diarrhea episodes should be supported in all low- and middle-income countries.

Analogues of Artificial Human Box C/D Small Nucleolar RNA As Regulators of Alternative Splicing of a pre-mRNA Target.

Small nucleolar RNAs (snoRNAs) play a key role in ribosomal RNA (rRNA) biogenesis. Box C/D snoRNAs guide the site-specific 2'-O-ribose methylation of nucleotides in rRNAs and small nuclear RNAs (snRNAs). A number of box C/D snoRNAs and their fragments have recently been reported to regulate post-transcriptional modifications and the alternative splicing of pre-mRNA. Artificial analogues of U24 snoRNAs directed to nucleotides in 28S and 18S rRNAs, as well as pre-mRNAs and mature mRNAs of human heat shock cognate protein (hsc70), were designed and synthesized in this study. It was found that after the transfection of MCF-7 human cells with artificial box C/D RNAs in complex with lipofectamine, snoRNA analogues penetrated into cells and accumulated in the cytoplasm and nucleus. It was demonstrated that the transfection of cultured human cells with artificial box C/D snoRNA targeted to pre-mRNAs induce partial splicing impairments. It was found that transfection with artificial snoRNAs directed to 18S and 28S rRNA nucleotides, significant for ribosome functioning, induce a decrease in MCF-7 cell viability.

Transcatheter closure of atrial septal defects--is balloon sizing still necessary?

INTRODUCTION: The device closure of atrial septal defects has evolved over the years. In the early days of transcatheter occlusion, balloon sizing was used to choose an appropriate sized device. We postulate that balloon sizing does not value-add to the procedure and is unnecessary. MATERIALS AND METHODS: Patients who had balloon sizing, with (Group 1, n = 38) or without (Group 2, n = 21) atrial septal defect closure, were compared to another group (Group 3, n = 64) who had atrial septal defect closure without balloon sizing. Although the atrial septal defect size (mm) in those without balloon sizing (Group 3) compared to patients who had balloon sizing (Group 1) (18.3 +/- 5.4 vs 14.8 +/- 5.8; P = 0.021) was larger, the Amplatzer septal occluder size chosen (mm) (21.6 +/- 6.3 vs 21.2 +/- 8.1; P = 0.693) was similar. RESULTS: We analysed the degree of absolute sizing, defined as [(Balloon or Amplatzer occluder size) - (transoesophageal echocardiography size)], versus relative sizing, which is defined as [(Balloon or Amplatzer occluder size)--(transoesophageal echocardiography size) / (Balloon or Amplatzer occluder size)]. It was evident that there was greater absolute and relative over-sizing (6.3 +/- 4.4 mm vs 4.2 +/- 2.1 mm; P = 0.009 and 28.3 +/- 15.4% vs 20.0 +/- 7.0%; P = 0.001, respectively) in patients with balloon sizing (Group 1) compared to those who did not (Group 3). Even a greater degree of absolute (5.1 +/- 3.9 mm vs 9.5 +/- 4.7 mm; P <0.001) and relative over-sizing (24.8 +/- 15.6% vs 33.0 +/- 13.6%; P = 0.001) was observed in patients who had balloon sizing but there was no closure (Group 2) compared to those who had balloon sizing and closure of their defects (Group 1). CONCLUSION: Our results showed that balloon sizing tended to over-size the atrial septal defect. This may have an important bearing in selecting a larger device than necessary, or even precluding transcatheter closure of the larger atrial septal defects. It is also associated with increased procedural, fluoroscopy time and cost. We suggest that balloon sizing may no longer be necessary in the protocol of device closure of an atrial septal defect.

Transcatheter closure of atrial septal defects--is balloon sizing still necessary?

<p><b>INTRODUCTION</b>The device closure of atrial septal defects has evolved over the years. In the early days of transcatheter occlusion, balloon sizing was used to choose an appropriate sized device. We postulate that balloon sizing does not value-add to the procedure and is unnecessary.</p><p><b>MATERIALS AND METHODS</b>Patients who had balloon sizing, with (Group 1, n = 38) or without (Group 2, n = 21) atrial septal defect closure, were compared to another group (Group 3, n = 64) who had atrial septal defect closure without balloon sizing. Although the atrial septal defect size (mm) in those without balloon sizing (Group 3) compared to patients who had balloon sizing (Group 1) (18.3 +/- 5.4 vs 14.8 +/- 5.8; P = 0.021) was larger, the Amplatzer septal occluder size chosen (mm) (21.6 +/- 6.3 vs 21.2 +/- 8.1; P = 0.693) was similar.</p><p><b>RESULTS</b>We analysed the degree of absolute sizing, defined as [(Balloon or Amplatzer occluder size) - (transoesophageal echocardiography size)], versus relative sizing, which is defined as [(Balloon or Amplatzer occluder size)--(transoesophageal echocardiography size) / (Balloon or Amplatzer occluder size)]. It was evident that there was greater absolute and relative over-sizing (6.3 +/- 4.4 mm vs 4.2 +/- 2.1 mm; P = 0.009 and 28.3 +/- 15.4% vs 20.0 +/- 7.0%; P = 0.001, respectively) in patients with balloon sizing (Group 1) compared to those who did not (Group 3). Even a greater degree of absolute (5.1 +/- 3.9 mm vs 9.5 +/- 4.7 mm; P <0.001) and relative over-sizing (24.8 +/- 15.6% vs 33.0 +/- 13.6%; P = 0.001) was observed in patients who had balloon sizing but there was no closure (Group 2) compared to those who had balloon sizing and closure of their defects (Group 1).</p><p><b>CONCLUSION</b>Our results showed that balloon sizing tended to over-size the atrial septal defect. This may have an important bearing in selecting a larger device than necessary, or even precluding transcatheter closure of the larger atrial septal defects. It is also associated with increased procedural, fluoroscopy time and cost. We suggest that balloon sizing may no longer be necessary in the protocol of device closure of an atrial septal defect.</p>

Blood serum immunoglobulins of patients with multiple myeloma are capable of hydrolysing histone H1.

BACKGROUND: Recently we have shown that the imunoglobulins G from blood serum of some multiple sclerosis patients are capable of cleaving histone H1. AIM: To check whether histone H1-hydrolyzing abzymes could be detected not only in blood plasma of autoimmune patients, but also during cancer development, particularly during the onset of multiple myeloma. METHODS: Immunoglobulines were isolated from blood serum of multiple myeloma patients (n = 11) by precipitation with 50% ammonium sulfate and tested for proteolytic activity toward linker and core calf thymus histones. Antibody preparations able to cleaved histone H1 were subjected to affinity chromatography on histone H1-Sepharose with following analysis of chromatographic fractions' protease activity. To prove that antibody molecules are responsible for hydrolysis of histone H1, gel filtration at acidic pH with subsequent examination of protease activity of chromatographic fractions (pH-shock analysis) was used. RESULTS: It was found that 3 of 11 antibody preparations are capable of hydrolyzing calf thymus histone H1 but not core histones. It was shown that histone H1-hydrolysing activity of 2 proteolytically active antibody preparations is associated with IgGs that possess affinity towards histone H1. pH-shock analysis proved that protease activity towards histone H1 is intrinsic property of IgG molecules. CONCLUSIONS: We demonstrated the existence of previously unknown histone H1 hydrolyzing IgG abzymes in the serum of multiple myeloma patients. Possible biological role of hisH1-hydrolyzing antibodies in patients with multiple myeloma was discussed.

3-Allyl-2-hydr-oxy-5,6,8-trimethoxy-naphthalene-1,4-dione.

In the crystal structure of the title compound, C(16)H(16)O(6), a pair of naphthoquinone rings are linked via O-H⋯O-C hydrogen bonds in a nearly orthogonal arrangement. This dimeric unit is linked to a neighbouring dimer by π-π stacking inter-actions between the naphthoquinone rings, where the distance between the mean plane of the naphtoquinone backbones is 3.468 Å, and O-H⋯O-C hydrogen bonds.

Secretory IgAs from human milk with affinity to mammalian DNA are capable of hydrolyzing ribosomal RNA.

It was found that milk of clinical healthy women contains sIgA possessing high affinity for the mammalian thymus DNA and DNA-hydrolyzing activity (sIgA-abzymes). Here we present data that such sIgA-abzymes, purified by sequential chromatography on DEAE-fractogel, heparin-sepharose, DNA-cellulose and followed by gel-filtration, are also able to hydrolyse total RNA from E. coli better than plasmid DNA. Besides, such sIgA-abzymes effectively cleaved 18S and 28S ribosomal RNA isolated from human A549 cells. It is noteworthy that the nuclease activity of sIgA-abzymes was significantly inhibited by ATP, while dATP had no effect on it. A potential role of the ribonuclease activity of sIgA-abzymes present in human milk is discussed.

Postnatal growth failure in preterm infants: ascertainment and relation to long-term outcome.

OBJECTIVE: Traditional measure of postnatal growth failure assessment has poor discriminatory power for long-term outcomes. Our objective was to identify measure of postnatal growth failure associated with long-term outcome in preterm infants born at < 28 weeks' gestation. PATIENTS AND METHODS: Four measures of defining postnatal growth failure at 36 weeks corrected gestational age: (1) weight < 10(th) centile, (2) weight < 3(rd) centile, (3) z score difference from birth > 1 and, (4) z score difference from birth > 2; were compared for their predictive values and strength of association with adverse neurodevelopmental outcomes at 18-24 months. RESULTS: Postnatal growth failure defined as a decrease in z score of > 2 between birth and 36 weeks corrected gestational age had the best predictive values compared to other postnatal growth failure measures, however, it was significantly associated with psychomotor developmental (P=0.006) but not with mental developmental indices (P=0.379). CONCLUSION: Postnatal growth failure defined by z score change influenced psychomotor but not mental tasks in this cohort. This method of ascertainment could be useful to identify infants who might benefit from nutritional interventions.

'He hath the French pox': stigma, social value and social exclusion.

Goods and resources are finite, and social forces heavily pattern their distribution. One of the principal mechanisms for shaping the distribution of resources is by regulating entitlement to community membership itself. By restricting groups' membership of community, so access to social goods and resources diminishes, which in turn has a negative impact on the health and wellbeing of the excluded groups. It is argued here that community membership is determined on the basis of the perceived social value of groups and individuals and stigmatisation is the marking of individuals and groups who are 'unworthy' of social investment. Using the notion of reciprocity we show how groups may be stigmatised and socially excluded as a mechanism for protecting limited social resources from exploitation. This perspective provides an empirically testable framework for the understanding of stigma and social exclusion that goes beyond the largely descriptive work that currently populates the field. We illustrate the process of stigmatisation and social exclusion and discuss how this suggests new styles of intervention, as well as new directions for research.