Anakinra in Sanfilippo syndrome: a phase 1/2 trial.

Sanfilippo syndrome is a fatal childhood neurodegenerative disorder involving neuroinflammation among multiple pathologies. We hypothesized that anakinra, a recombinant interleukin-1 receptor antagonist, could improve neurobehavioral and functional symptoms owing to its capacity to treat neuroinflammation. This phase 1/2 trial aimed to test the safety, tolerability and effects of anakinra on neurobehavioral, functional and quality-of-life outcomes in patients and their caregivers. The primary outcome was the percent of participants requiring a dose increase at week 8 or week 16. Secondary efficacy outcomes included a multi-domain responder index (MDRI). Twenty-three participants (6-26 years of age) were enrolled. Twenty continued treatment to week 8, and 15 (75%) required an increased dose at week 8 or week 16. There was an improvement in at least one domain in the MDRI in 18 of 21 (86%) at week 8 and in 15 of 16 (94%) at week 36. Seven participants withdrew (intolerability of daily injections and lost to follow-up) before week 36. Adverse events occurred in 22 of 23 (96%) participants, most commonly mild injection site reactions. No serious adverse events were related to anakinra. In conclusion, anakinra was safe and associated with improved neurobehavioral and functional outcomes, supporting continued investigation of anakinra in Sanfilippo syndrome and other mucopolysaccharidoses. ClinicalTrials.gov identifier: NCT04018755 .

The immune landscape of uterine fibroids as determined by mass cytometry.

OBJECTIVE: To study the differences in immune cell profiles in uterine fibroids (Fibs) and matched myometrium (Myo). DESIGN: Observational study. SETTING: Laboratory study. PATIENT(S): The study included tissue that was collected from 10 pairs of Fib and matched Myo from women, not on hormonal medications, undergoing hysterectomy and myomectomy. INTERVENTION(S): None. MAIN OUTCOME MEASURE(S): Differences in immune cell and cytokine composition between Fib and matched Myo. RESULT(S): The mass cytometry analysis indicated that Fibs had a significantly higher number of natural killer (NK) cells, total macrophages, M2 macrophages, and conventional dendritic cells when compared with matched Myo from the same patient. In contrast, Fibs had significantly fewer CD3 and CD4 T cells when compared with Myo. The mass cytometry analysis results did not show any significant difference in the number of resting mast cells. Immunoflurorescent and immunohistochemical imaging confirmed the cytometry by time of flight results, showing a significantly higher number of NK cells, tryptase-positive mast cells indicative of mast cell activation, total macrophages, and M2 cells in Fibs and a significantly lower number of CD3 and CD4 T cells. The cytokine assay revealed significantly increased levels of human interferon α2, interleukin (IL)-1α, and platelet-derived growth factor AA and significantly lower levels of macrophage colony-stimulating factor and IL-1 receptor antagonist in Fib. CONCLUSION(S): Our results show significant differences in immune cell populations and cytokine levels between Fib and Myo. These differences could account for the increased inflammation in fib and a potential mechanism by which these tumors evade the immune system. These findings provide a foundation for further studies exploring the role of immune cells in Fib development.

Skeletal muscle cell protein dysregulation highlights the pathogenesis mechanism of myopathy-associated p97/VCP R155H mutations.

p97/VCP, a hexametric member of the AAA-ATPase superfamily, has been associated with a wide range of cellular protein pathways, such as proteasomal degradation, the unfolding of polyubiquitinated proteins, and autophagosome maturation. Autosomal dominant p97/VCP mutations cause a rare hereditary multisystem disorder called IBMPFD/ALS (Inclusion Body Myopathy with Paget's Disease and Frontotemporal Dementia/Amyotrophic Lateral Sclerosis), characterized by progressive weakness and subsequent atrophy of skeletal muscles, and impacting bones and brains, such as Parkinson's disease, Lewy body disease, Huntington's disease, and amyotrophic lateral ALS. Among all disease-causing mutations, Arginine 155 to Histidine (R155H/+) was reported to be the most common one, affecting over 50% of IBMPFD patients, resulting in disabling muscle weakness, which might eventually be life-threatening due to cardiac and respiratory muscle involvement. Induced pluripotent stem cells (iPSCs) offer an unlimited resource of cells to study pathology's underlying molecular mechanism, perform drug screening, and investigate regeneration. Using R155H/+ patients' fibroblasts, we generated IPS cells and corrected the mutation (Histidine to Arginine, H155R) to generate isogenic control cells before differentiating them into myotubes. The further proteomic analysis allowed us to identify differentially expressed proteins associated with the R155H mutation. Our results showed that R155H/+ cells were associated with dysregulated expression of several proteins involved in skeletal muscle function, cytoskeleton organization, cell signaling, intracellular organelles organization and function, cell junction, and cell adhesion. Our findings provide molecular evidence of dysfunctional protein expression in R155H/+ myotubes and offer new therapeutic targets for treating IBMPFD/ALS.

Brain transplantation of genetically corrected Sanfilippo type B neural stem cells induces partial cross-correction of the disease.

Sanfilippo syndrome type B (mucopolysaccharidosis type IIIB) is a recessive genetic disorder that severely affects the brain due to a deficiency in the enzyme α-N-acetylglucosaminidase (NAGLU), leading to intra-lysosomal accumulation of partially degraded heparan sulfate. There are no effective treatments for this disorder. In this project, we carried out an ex vivo correction of neural stem cells derived from Naglu -/- mice (iNSCs) induced pluripotent stem cells (iPSC) using a modified enzyme in which human NAGLU is fused to an insulin-like growth factor II receptor binding peptide in order to improve enzyme uptake. After brain transplantation of corrected iNSCs into Naglu -/- mice and long-term evaluation of their impact, we successfully detected NAGLU-IGFII activity in all transplanted animals. We found decreased lysosomal accumulation and reduced astrocytosis and microglial activation throughout transplanted brains. We also identified a novel neuropathological phenotype in untreated Naglu -/- brains with decreased levels of the neuronal marker Map2 and accumulation of synaptophysin-positive aggregates. Upon transplantation, we restored levels of Map2 expression and significantly reduced formation of synaptophysin-positive aggregates. Our findings suggest that genetically engineered iNSCs can be used to effectively deliver the missing enzyme to the brain and treat Sanfilippo type B-associated neuropathology.

Notch activation is required for downregulation of HoxA3-dependent endothelial cell phenotype during blood formation.

Hemogenic endothelium (HE) undergoes endothelial-to-hematopoietic transition (EHT) to generate blood, a process that requires progressive down-regulation of endothelial genes and induction of hematopoietic ones. Previously, we have shown that the transcription factor HoxA3 prevents blood formation by inhibiting Runx1 expression, maintaining endothelial gene expression and thus blocking EHT. In the present study, we show that HoxA3 also prevents blood formation by inhibiting Notch pathway. HoxA3 induced upregulation of Jag1 ligand in endothelial cells, which led to cis-inhibition of the Notch pathway, rendering the HE nonresponsive to Notch signals. While Notch activation alone was insufficient to promote blood formation in the presence of HoxA3, activation of Notch or downregulation of Jag1 resulted in a loss of the endothelial phenotype which is a prerequisite for EHT. Taken together, these results demonstrate that Notch pathway activation is necessary to downregulate endothelial markers during EHT.

HIV-1 Tat induces DNMT over-expression through microRNA dysregulation in HIV-related non Hodgkin lymphomas.

BACKGROUND: A close association between HIV infection and the development of cancer exists. Although the advent of highly active antiretroviral therapy has changed the epidemiology of AIDS-associated malignancies, a better understanding on how HIV can induce malignant transformation will help the development of novel therapeutic agents. METHODS: HIV has been reported to induce the expression of DNMT1 in vitro, but still no information is available about the mechanisms regulating DNMT expression in HIV-related B-cell lymphomas. In this paper, we investigated the expression of DNMT family members (DNMT1, DNMT3a/b) in primary cases of aggressive B-cell lymphomas of HIV-positive subjects. RESULTS: Our results confirmed the activation of DNMT1 by HIV in vivo, and reported for the first time a marked up-regulation of DNMT3a and DNMT3b in HIV-positive aggressive B-cell lymphomas. DNMT up-regulation in HIV-positive tumors correlated with down-regulation of specific microRNAs, as the miR29 family, the miR148-152 cluster, known to regulate their expression. Literature reports the activation of DNMTs by the human polyomavirus BKV large T-antigen and adenovirus E1a, through the pRb/E2F pathway. We have previously demonstrated that the HIV Tat protein is able to bind to the pocket proteins and to inactivate their oncosuppressive properties, resulting in uncontrolled cell proliferation. Therefore, we focused on the role of Tat, due to its capability to be released from infected cells and to dysregulate uninfected ones, using an in vitro model in which Tat was ectopically expressed in B-cells. CONCLUSIONS: Our findings demonstrated that the ectopic expression of Tat was per se sufficient to determine DNMT up-regulation, based on microRNA down-regulation, and that this results in aberrant hypermethylation of target genes and microRNAs. These results point at a direct role for Tat in participating in uninfected B-cell lymphomagenesis, through dysregulation of the epigenetical control of gene expression.

EBV Reactivation and Chromosomal Polysomies: Euphorbia tirucalli as a Possible Cofactor in Endemic Burkitt Lymphoma.

Burkitt lymphoma is endemic in the Equatorial Belt of Africa, its molecular hallmark is an activated, MYC gene mostly due to a chromosomal translocation. Especially in its endemic clinical variant, Burkitt lymphoma is associated with the oncogenic Epstein-Barr virus (EBV), and holoendemic malaria acts as an amplifier. Environmental factors may also cooperate in Burkitt lymphomagenesis in the endemic regions, such as plants used as traditional herbal remedies. Euphorbia tirucalli, a plant known to possess EBV-activating substances, has a similar geographical distribution to endemic Burkitt's Lymphoma and is used as a hedge, herbal remedy and toy in the Lymphoma BeltI. In this study we aimed at determining if exposure to Euphorbia tirucalli could contribute to lymphomagenesis, and at which extent. Lymphoblastoid and cord blood-derived cell lines were treated with plant extracts, and the expression of EBV-coded proteins was checked, to assess EBV reactivation. The occurrence of chromosomal translocations was then investigated by FISH. Our preliminary results suggest that E. tirucalli is able to reactivate EBV and determine chromosomal alterations, which leads to c-MYC altered expression. The existence of genomic alterations might determine the accumulation of further genetic alteration, which could eventually lead to a transformed phenotype.

Downregulation and aberrant promoter methylation of p16INK4A: a possible novel heritable susceptibility marker to retinoblastoma.

RB loss has long been recognized as the causative genetic alteration underlying retinoblastoma but it is increasingly evident that other alterations are required for the tumor to develop. Therefore, we set out to identify additional inheritable susceptibility markers and new potential preventive and therapeutic targets for retinoblastoma. We focused on the p16INK4A tumor suppressor gene because of its possible role in retinoblastoma pathogenesis and its involvement in predisposition to familial cancer. p16INK4A expression was analyzed in tumor samples from retinoblastoma patients by immunohistochemistry and in peripheral blood cells from both patients and their parents by real-time quantitative reverse transcription-PCR (qRT-PCR). Since promoter methylation is a common mechanism regulating p16INK4A expression, the methylation status of its promoter was also analyzed in blood samples from patients and their parents by methylation-specific PCR. A downregulation of p16INK4A was observed in 55% of retinoblastoma patients. Interestingly, in 56% of the cases showing p16INK4A downregulation at least one of the patients' parents bore the same alteration in blood cells. Analysis of p16INK4A promoter methylation showed hypermethylation in most patients with p16INK4A downregulation and in the parents with the same alteration in p16INK4A expression. The finding that p16INK4A was downregulated both in patients and their parents suggests that this alteration could be a novel inheritable susceptibility marker to retinoblastoma. The observation that p16INK4A downregulation seems to be due to its promoter hypermethylation opens the way for the development of new preventive and therapeutic strategies using demethylating agents.

Geographic variation and environmental conditions as cofactors in Chlamydia psittaci association with ocular adnexal lymphomas: a comparison between Italian and African samples.

A particular extra-nodal lymphoma type arises from B cells of the marginal zone (MZ) of mucosa-associated lymphoid tissue (MALT). The aetiology of MZ lymphomas suggests that they are associated with chronic antigenic stimulation by microbial pathogens, among which Helicobacter pylori-associated gastric MALT lymphoma is the best studied. Recently, MALT lymphomas have been described in the context of chronic conjunctivitis, which can be associated with Chlamydia spp. infection. Studies from Italy showed the presence of Chlamydia psittaci in 87% of ocular adnexal lymphomas (OAL), and C. psittaci has been described in a large part of samples from Austria and Korea as well. However, this finding was not always confirmed by other studies, suggesting that the association with C. psittaci may depend on geographic heterogeneity. Interestingly, none of the studies up to now has been carried out in the African population, where a strong association between infectious agents and the occurrence of human neoplasms has been reported. This study was designed to investigate the possible association of Chlamydia psittaci in cases retrieved from Kenya, compared to cases from Italy. Our results showed that there was a marked variation between the two geographical areas in terms of association with C. psittaci, as 17% (5/30) of the samples from Italy were positive for C. psittaci, whereas no association with this pathogen was observed in any of the African samples (0/9), suggesting that other cofactors may determine the OAL occurrence in those areas. OAL cases are often characterized by down-regulation of p16/INK4a expression and promoter hypermethylation of the p16/INK4a gene. Our results showed a partial methylation of p16/INK4a promoter in C. psittaci-negative cases, whereas no hypermethylation of this gene was found in C. psittaci-positive cases, suggesting that mechanisms other than promoter hypermethylation lead to p16/INK4a silencing in C. psittaci-positive cases. We may conclude that the role of epidemiologic, environmental and genetic factors, must be considered in the aetiology of this disease.

The role of the Cdk9/Cyclin T1 complex in T cell differentiation.

The Cdk9/Cyclin T1 complex is very important in controlling specific differentiative pathways of several cell types, including muscle cells and neurons. We recently demonstrated the involvement of this complex in B cell activation/differentiation. To check whether the Cdk9/Cyclin T1 complex is also involved in the T cell activation/differentiation process, we isolated different T cell populations by magnetic separation, based on their surface antigens. We observed that the expression level of Cdk9/Cyclin T1 increases in effector T cells (CD27(+)), as well as in activated T cells (CD25(+)) and memory T cells (CD45RA(-)), thus suggesting a specific upregulation of the Cdk9/Cyclin T1 complex following antigen encounter. We have previously demonstrated that in B cells, Cdk9 interacts in vivo with the E2A gene products E12/E47 (members of the basic helix-loop-helix family) which are involved in differentiation. In this article, we show that this interaction also occurs in T cells. This suggests an active role for the Cdk9/Cyclin T1 complex during lymphoid differentiation, through physical binding with E12 and E47. These preliminary results suggest that the Cdk9/Cyclin T1 complex may be important in the activation and differentiation program of lymphoid cells and that its upregulation, which is due to still unknown mechanisms, may contribute to malignant transformation.

Defining episodes of diarrhoea: results from a three-country study in Sub-Saharan Africa.

The study was conducted to assess the effect of definition of episode on diarrhoeal morbidity and to develop a means of adjusting estimates of morbidity for the definition of episode used. This paper reports on a cohort study of 374 children, aged 9-32 months, in three African countries, which recorded frequency and consistency of stool over a seven-month period. Different definitions of episode were applied to these data to assess their effect on annualized diarrhoeal morbidity. Adjustment factors were then derived that corrected morbidity for non-standard definitions of episode. Applying non-standard definitions of episode gave estimates of an annualized number of episodes between 38% and 137% of the internationally-accepted definition. Researchers should be encouraged to use the standard definition of episode of diarrhoea and to use appropriate field protocols. Where this is not possible, correction factors should be applied, particularly where estimates of diarrhoeal morbidity are pooled in systematic reviews.

The conceptual framework for estimating food energy requirement.

OBJECTIVE: In anticipation of the revision of the 1985 Food and Agricultural Organization/World Health Organization/United Nations University (FAO/ WHO/UNU) Expert Consultation Report on 'Energy and Protein Requirements', recent scientific knowledge on the principles underlying the estimation of energy requirement is reviewed. DESIGN: This paper carries out a historical review of the scientific rationale adopted by previous FAO/WHO technical reports on energy requirement, discusses the concepts used in assessing basal metabolic rate (BMR), energy expenditure, physical activity level (PAL), and examines current controversial areas. Recommendations and areas of future research are presented. CONCLUSIONS: The database of the BMR predictive equations developed by the 1985 FAO/WHO/UNU Expert Consultation Report on Energy and Protein Requirements needs updating and expansion, applying strict and transparent selection criteria. The existence of an ethnic/tropical factor capable of affecting BMR is not supported by the available evidence. The factorial approach for the calculation of energy requirement, as set out in the 1985 report, should be retained. The estimate should have a normative rather than a prescriptive nature, except for the allowance provided for extra physical activity for sedentary populations, and for the prevention of non-communicable chronic diseases. The estimate of energy requirement of children below the age of 10 years should be made on the basis of energy expenditure rather than energy intake. The evidence of the existence of an ethnic/tropical factor is conflicting and no plausible mechanism has as yet been put forward.

Питание и здоровье в Европе: новая основа для действий

В данной публикации, предназначенной для работников здравоохранения, всесторонне рассматриваются различные аспекты политики в области пищевых продуктов и питания и приводятся соответствующие фактические данные. В ней затрагиваются вопросы ухудшения состояния здоровья, вызванного пищевыми продуктами и питанием, и связанных с этим затрат, обосновывается необходимость принятия мер и предлагаются конкретные действия, которые могут предприниматься лицами, определяющими политику. Особое внимание в публикации уделяется насущной необходимости выработки комплексной политики в области пищевых продуктов и питания с участием различных секторов в целях стабильного и непрерывного производства продуктов питания, их безопасности и обеспечения всех людей пищевыми продуктами высокого качества и питательной ценности. Краткое содержание данной публикации для лиц, разрабатывающих и определяющих политику, опубликовано в 2002 г. на англ. яз. и рус. яз. Плохое питание, болезни, передаваемые через пищевые продукты, и отсутствие безопасного доступа к доброкачественным пищевым продуктам являются весьма важной составляющей бремени болезней и смертности в Европейском регионе ВОЗ. Улучшение рациона и режима питания, безопасность пищевых продуктов и обеспеченность продуктами питания не только уменьшат или предотвратят страдания отдельных людей и населения в целом, но и помогут сократить расходы и издержки систем здравоохранения и обеспечить социально-экономические выгоды и преимущества для стран.

Food and health in Europe: a new basis for action.

Poor nutrition, foodborne disease and lack of secure access to good food make an important contribution to the burden of disease and death in the WHO European Region. Better diets, food safety and food security will not only reduce or prevent suffering to individuals and societies but also help cut costs to health care systems and bring social and economic benefits to countries. People's chances for a healthy diet depend less on individual choices than on what food is available and whether it is affordable. Policies to benefit health through good food and nutrition must extend beyond the health sector to include sectors ranging from agriculture and food processing, manufacturing and trade to transport, retailing, catering and advertising. Food and nutrition policies should be coordinated so that public health is given due priority in the making of food policies by non-health sectors. This publication discusses in depth the components of food and nutrition policies and the evidence supporting them. It describes food- and nutrition-related ill health and its costs, shows the need for action and describes the steps for decision-makers to take. This book highlights the urgent need for integrated, multisectoral food and nutrition policies to encourage the sustainable production of food, its safety and the provision of food of high nutritional quality for all.

Food and health in Europe: a new basis for action

Aimed at health professionals, this publication discusses in depth the components of food and nutrition policies and the evidence supporting them. It describes food- and nutrition-related ill health and its costs, shows the need for action and describes the steps for decision-makers to take. This book highlights the urgent need for integrated, multisectoral food and nutrition policies to encourage the sustainable production of food, its safety and the provision of food of high nutritional quality for all. A summary of the book’s content, aimed at policy-makers, was published in English and Russian in 2002. Poor nutrition, foodborne disease and lack of secure access to good food make an important contribution to the burden of disease and mortality in the WHO European Region. Better diets, food safety and food security will not only reduce or prevent suffering to individuals and societies but also help cut costs to health care systems and bring social and economic benefits to countries.

Capacidad antioxidante y potencial de sinergismo entre los principales constituyentes antioxidantes de algunos alimentos / Antioxidant capacity and synergistic potential among the main antioxidant components of some food

Los objetivos de este estudio fueron: evaluar la capacidad antioxidante de alimentos seleccionados (tomate, cebolla, lechuga y col), su efecto protector sobre la peroxidación de ácido linoleico, e investigar el potencial de interacción entre diferentes antioxidantes como flavonoides (rutina y quercetina), ácidos fenólicos (ácido cafeico) y vitaminas (vitamina E y C). Se examinó la eficiencia antioxidante por 2 sistemas: la capacidad antioxidante total y el efecto protector sobre la peroxidación del ácido linoleico. La mayor capacidad antioxidante total la presentó la col, seguida por la lechuga, cebolla y tomate y el mayor efecto protector sobre la peroxidación del ácido linoleico también lo presentó la col, seguida por el tomate, la lechuga y la cebolla. En el estudio de interacción se obtuvo una variedad de respuestas, desde un antagonismo hasta un sinergismo. En conclusión, los extractos de vegetales frescos muestran un efecto antioxidante diferente y su actividad depende de la naturaleza y concentración de los antioxidantes naturales presentes en el alimento. El uso de 2 sistemas (hidrofílico y lipofílico) es fundamental para estudiar la capacidad antioxidante de un compuesto o de un extracto de alimento (AU)

Capacidad antioxidante y potencial de sinergismo entre los principales constituyentes antioxidantes de algunos alimentos / Antioxidant capacity and synergistic potential among the main antioxidant components of some food

Los objetivos de este estudio fueron: evaluar la capacidad antioxidante de alimentos seleccionados (tomate, cebolla, lechuga y col), su efecto protector sobre la peroxidación de ácido linoleico, e investigar el potencial de interacción entre diferentes antioxidantes como flavonoides (rutina y quercetina), ácidos fenólicos (ácido cafeico) y vitaminas (vitamina E y C). Se examinó la eficiencia antioxidante por 2 sistemas: la capacidad antioxidante total y el efecto protector sobre la peroxidación del ácido linoleico. La mayor capacidad antioxidante total la presentó la col, seguida por la lechuga, cebolla y tomate y el mayor efecto protector sobre la peroxidación del ácido linoleico también lo presentó la col, seguida por el tomate, la lechuga y la cebolla. En el estudio de interacción se obtuvo una variedad de respuestas, desde un antagonismo hasta un sinergismo. En conclusión, los extractos de vegetales frescos muestran un efecto antioxidante diferente y su actividad depende de la naturaleza y concentración de los antioxidantes naturales presentes en el alimento. El uso de 2 sistemas (hidrofílico y lipofílico) es fundamental para estudiar la capacidad antioxidante de un compuesto o de un extracto de alimento