New forms of induction of apoptosis in aggressive lymphoma using peptides that interrupt the RAS / RAF interaction

Interpretation: RAS-RAF-MEK-ERK is a key pathway for apoptosis regulation in cancer cells. B-Raf-inhibitors such as PLX4032 peptide was developed by Institute Curie-Université Pierre et Marie Curie in order to induce apoptosis in cancer cells. Objectives: To demonstrate pro-apoptotic properties and survival outcome of EP2014/064243 peptide in murine aggressive lymphoma. Material and Methodology: BALBc mice with T-lymphoma were randomized assigned either in Group A (peptide+cyclophosphamide-CFM); Group B (peptides), Group C (CFM-control) or Control D (Cl-Na 0.9%-SF control group). Survival probability was calculated by Kaplan-Meier analysis. Apoptosis was detected using TUNEL technique. The protocol was approved by the Institutional Committee for Animal Care (CICUAL: T04-01-2015) Results: The median survival was 24 days (21.6-26.4) for placebo, 33 days (28.0-35.4) for the CFM monotherapy group, 33 (27.1-35.8) for the peptide group and 34 days (24,4-40) for CFM-peptide combined treatment (p<0.05). In lymph node tissue the mean TUNEL positive cells per field for each treatment group was 2, 12 and 13 and 35 for SF, CFM, peptide and combined therapy (p<0.05). Conclusion: These findings suggest that in murine aggressive lymphoma treated by an experimental peptide in addition with CFM, had an exponentially pro-apoptotic effect than CFM alone, suggesting that the peptide potentiated the anti-tumoural effect of CFM.

Correction to: T cell leukemia control via Ras-Raf pathway inhibition with peptides.

[This corrects the article on p. 172 in vol. 10, PMID: 29075346.].

T cell leukemia control via Ras-Raf pathway inhibition with peptides.

RATIONALE: RAS-RAF-MEK-ERK pathway has been considered a promising target for anticancer therapy. However, tumor cells may develop resistance against such drugs via hyperactivation of N-Ras, which explains why novel therapeut-ic approaches. In this sense, the Institute Curie- Université Pierre et Marie Curie (Paris 6) designed peptides in order to disturb Ras/Raf interaction which showed pro-apoptotic properties. These peptides were patented as WO2015001045 A2 (PCT/EP2014/064243)5. OBJECTIVE: In order to check the anti-tumoral action of WO2015001045 A2 peptides in a very aggressive BALB/c mice spontaneous leukemia called LB, we performed the present study. METHOD & RESULTS: 50 BALB/c mice inoculated with 106 LB tumor cells were randomly assigned either to control (placebo) or treatment group (that daily received 3 mg of peptide per kg of mice) during 30 days. By day 15 only 24% of the control group was alive vs. 100% of the treatment group. The average survival in treated group was 20,27 days while in control group the mean survival was 15,48 days. Either bone marrow, spleen or axillary nodes demonstrated a higher level of malignant T cell presence compare with treated group (89,78% ; 95,64% & 77,68% versus 72,45%, 80,23% & 63.44% respectively for each organ inspected. DISCUSSION: Our study demonstrated an improvement in survival curves in mice model affected by spontaneous T lymphoid leukemia when peptides WO2015001045 A2 were used. These peptides might be a valid option to become part of the therapeutic armory for malignant lymphoproliferative diseases control.

Personalised iron supply for prophylaxis and treatment of pregnant women as a way to ensure normal iron levels in their breast milk.

RATIONALE: Because the characteristics of all body fluids depends on patient's health status, is it possible that disadvantaged and socially vulnerable mothers may have lower amounts of iron in their breast milk, and that their babies receive lower content of the mineral for their normal growth and development. Assuring a preventive treatment of the mother might solve this problem. OBJECTIVE: To demonstrate breast milk iron content from disadvantaged mothers and impact of personalized iron supplementation program. MATERIALS AND METHODS: cross-sectional study. Breast milk samples were obtained for ferritin analysis. Health's services usually provides free folic acid and iron treatment however, treatment compliance is low. Patients were random in two groups: "A: Controls" that had free iron tablets available from Health Centre; and "B: Intervention" group where patients accepted to be periodically contacted at home by health's team for personalized iron dispensation. RESULTS: 360 patients were included. Profilaxis and treatment compliance were 100% and 97,6% for B group while for "Control" one was 63% and 34%(p0.0001). Higher breast milk iron levels were detected in Intervention's mothers compared with control's patients (p0.007). CONCLUSION: Personalized iron prophylaxis and treatment increased breast milk iron levels. Public health policy must ensure iron dispensation for each underserved mother in order to reduce children problems associate to iron deficiency during the first year of their life.

Outstanding survival and regeneration process by the use of intelligent acellular dermal matrices and mesenchymal stem cells in a burn pig model.

A pig model with a deep large burn was used to study the regeneration process induced by mesenchymal stem cells (MSCs) and acellular pig dermal matrices, made intelligent by the combination with biodegradable nanofibers loaded with growth factors (granulocyte-macrophage colony-stimulating factor and epidermal growth factor) and coated with the anti-CD44 monoclonal antibody (intelligent acellular dermal matrices, IADMs). These IADMs are specially designed to integrate in the wound bed as new biological scaffolds as well as to specifically recruit and attach circulating and/or externally applied MSCs through the anti-CD44 antibody while delivering precise amounts of growth factors. In this way, the reparative process as well as the aesthetic and functional results were enhanced in our burn model. The animal survived, the wound was completely closed, and total regeneration of the skin was obtained without much scarring. Surprisingly, hair follicles and other skin appendages developed despite the severity and deepness of the burn. Even burned muscles and ribs seemed to have undergone a regenerative process by the end of the study. Based on these findings, we have proposed the use of IADMs and autologous, allogeneic or xenogeneic MSCs, as a new paradigm for the future treatment of large burns and probably other dermatological and cosmetic human conditions.

The next generation of burns treatment: intelligent films and matrix, controlled enzymatic debridement, and adult stem cells.

We describe a novel technology based on nanoengineered multifunctional acellular biologic scaffolds combined with wound dressings and films of the same kind. This method allows selective delivery and release of shielded biomaterials and bioactive substances to a desired wound or damaged tissue while stimulating the selective anchoring and adhesion of endogenous circulating repairing cells, such as mesenchymal stem cells, to obtain a faster and more physiologic healing process. We also present a new controlled enzymatic debridement process for more effective burned tissue scarolysis. In light of our preliminary in vitro and in vivo data, we are convinced that these approaches can include the use of other kinds of adult stem cells, such as endometrial regenerative cells, to improve the vascularization of the constructs, with great potential in the entire tissue and organ regeneration field but especially for the treatment of severely burned patients, changing the way these lesions may be treated in the future.

Matrix superhighways configurations: new concepts for complex organ regeneration.

New ideas and experimental models for tissue and organ regeneration are urgently needed. There are several exciting challenges in the field of organogenesis that need to be defined. The integrated signals and molecular repertoires that shape the particular architecture of specific organs like the kidney or the liver are not completely understood yet. To develop a new scientific platform to be able to build up complex organs we have established a research program using basically Acellular Xenogeneic Isomorphic Matrices (AXIMs) and mesenchymal stem cells (MSCs) generating the necessary concepts for the definition, production, and application of the specific configurations of these matrices for organ regeneration. New and interesting pathways for MSC differentiation were identified. We believe that all extracellular matrices were created fundamentally equal or at least very similar in nature. We also believe that there are true "matrix superhighway configurations" with different three-dimensional geometrical architectures as well as biochemical, electrical, and molecular properties that are tissue and organ specific that influence cell differentiation and organogenesis and will be fundamental for the in vitro regeneration of complex organs for transplantation.

Bloodstream cells phenotypically identical to human mesenchymal bone marrow stem cells circulate in large amounts under the influence of acute large skin damage: new evidence for their use in regenerative medicine.

OBJECTIVES: Recent work has shown that human bone marrow contains mesenchymal stem cells (MSCs). However, little is known about their presence in peripheral blood. Since these cells are potentially responsible for tissue repair after injury, their number should be increased during these situations. To demonstrate their number during these situations, we measured MSCs in the peripheral blood of healthy donors and burn patients. MATERIALS AND METHODS: Blood samples were obtained from 15 acute burn patients and 15 healthy donors. We performed flow cytometric analysis, using a large monoclonal antibody panel: CD44, CD45, CD14, DR, CD34, CD19, CD13, CD29, CD105, CD1a, CD90, CD38, CD25. MSC phenotype was considered positive for CD44, CD13, CD29, CD90, and CD105, and negative for the other monoclonals. The testing was performed on day 3 after injury. We correlated the results with the age, sex, and size and type of burns. RESULTS: Cells expressing the MSC phenotype were detected in the peripheral blood of both groups. Noteworthy, compared with samples from healthy donors (0.0078 +/- 0.0044), blood obtained from burn patients showed a higher MSC percentage (0.1643 +/- 0.115; P < .001). The percentage of MSCs correlated with the size and severity of the burn. Increased values were also observed among younger patients. CONCLUSIONS: MSCs have an important role in regenerative processes of human tissues. We found cells phenotypically identical to MSCs circulating in physiological number in normal subjects, but in significantly higher amounts during acute large burns. Therefore, they may represent a previously unrecognized circulatory component to the process of skin regeneration.

[Interdisciplinary health project: a model based in home care makes the difference]. / Proyecto de acción interdisciplinaria en salud (PAIS): un modelo de atención centrado en el hogar hace la diferencia.

OBJECTIVE: To propose an interdisciplinary approach in Community Health through a model based on Home Health Care. DESIGN: A prospective study programmed in two periods: a cross sectional study for data collection followed by home assistance for the health problems detected. METHODS: 100 families (570 inhabitants) were randomly submitted to a clinical, nutritional and laboratory examination in order to detect major health problems. Afterwards the same professional team took care of 10 families in health promotion counseling and illness treatment. RESULTS: This study shows that 84% of the patients in whom a health abnormality was detected were not aware of their illness. The population was mainly affected by hypertension, hypercholesterolemia, hypertrigliceridemia, and anemia. None of these problems were listed as the main causes of consultation in a local center belonging to the traditional health services. Almost 70% of the inhabitants consumed a daily diet with less nutrients and calories than recommended. The project included health care for all family members and free drugs provision for all illnesses detected. Ninety-eight percent of patients had a proper and continuous follow up of their illness for a 3 year period, compared with a 31% follow up provided by the traditional health care organization. CONCLUSIONS: This work shows an integrated effort by the University, health centers, and all health care disciplines to propose an alternative program that personalizes attention and moves health care from Institutions to people's own homes. This model improved community health conditions, had a high impact on illness follow up and increased health care accessibility.

Proyecto de acción interdisciplinaria en salud (PAIS): un modelo de atención centrado en el hogar hace la diferencia / Interdisciplinary health project: a model based in home care makes the difference

Objetivo. Proponer un abordaje interdisciplinario de la atención de la salud centrado en los hogares y comparar la morbilidad detectada mediante dicho enfoque, con aquella objetivada en el sistema sanitario tradicional. Materiales y métodos. Estudio prospectivo con diagnóstico de situación (mediante encuestas socioeconómicas, nutricionales y examen físico); y posterior acción comunitaria sobre 100 hogares de la zona (570 personas), elegidos en forma aleatoria. Resultados. Los datos mostraron una población afectada por enfermedades como hipertensión arterial, hipercolesterolemia, anemia y malnutrición; mientras que el modelo sanitario tradicional registró como prevalentes otras enfermedades totalmente diferentes a las anteriores. Además, el 84% de los individuos con alguna patología, no percibió la enfermedad.Después del diagnóstico inicial, se realizó una acción continua, en los planos de promoción de la salud, educación, atención y monitoreo de las patologías detectadas y fabricación y dispensación gratuita de medicamentos para tratamientos completos durante 36 meses. El seguimiento de los pacientes se cumplió en un 98% durante dicho período, en comparación con el 31% logrado por el sistema sanitario tradicional.Conclusiones. En este trabajo se muestra a un grupo interdisciplinario en una propuesta alternativa para la atención de la salud, la cual traslada el eje desde los servicios instalados, que esperan pasivamente ser utilizados, a una más activa que pone en el centro los propios hogares, basando las acciones en las necesidades, logrando tener un mapa real de las patologías prevalentes, incrementando la accesibilidad a sus controles periódicos

Objective. To propose an interdisciplinary approach in Community Health through a model based on Home Health Care. Design. A prospective study programmed in two periods: a cross sectional study for data collection followed by home assistance for the health problems detected. Methods. 100 families (570 inhabitants) were randomly submitted to a clinical, nutritional and laboratory examination in order to detect major health problems. Afterwards the same professional team took care of 10 families in health promotion counseling and illness treatment. Results. This study shows that 84% of the patients in whom a health abnormality was detected were not aware of their illness. The population was mainly affected by hypertension, hypercholesterolemia, hypertrigliceridemia, and anemia. None of these problems were listed as the main causes of consultation in a local center belonging to the traditional health services. Almost 70% of the inhabitants consumed a daily diet with less nutrients and calories than recommended. The project included health care for all family members and free drugs provision for all illnesses detected. Ninety-eight percent of patients had a proper and continuous follow up of their illness for a 3 year period, compared with a 31% follow up provided by the traditional health care organization. Conclusions. This work shows an integrated effort by the University, health centers, and all health care disciplines to propose an alternative program that personalises attention and moves health care from Institutions to people’s own homes. This model improved community health conditions, had a high impact on illness follow up and increased health care accessibility

Human mesenchymal stem cells are tolerized by mice and improve skin and spinal cord injuries.

INTRODUCTION: We sought to use human mesenchymal stem cells (HMSC) for skin and spinal cord repair in mice. MATERIALS AND METHODS: Human bone marrow obtained from a young healthy donor was used to separate and culture human mesenchymal stem cells (HMSC). Ten mice were included in each of four groups. A full-thickness skin defect was surgically performed on all mice in groups 1 and 2. A transverse complete medullar section was performed in groups 3 and 4. Groups 1 and 3 received HMSC IV infusion and local HMSC polymer implant. Groups 2 and 4 received only the IV HMSC infusion. Five control animals from each group went through the same lesions but they didn't receive treatment. RESULTS: After local administration of HMSC into the fibrin polymer combined with the IV infusion of HMSC, there was no immune rejection; all skin defects healed without scar or retraction at a median time of 14 days. Sixty percent of the animals treated with IV infusion and polymer with HMSC simultaneously had improved neurological activities, while all control mice with spinal cord injury experiments died or perpetuated their paralysis with worsening muscular atrophy and increasing propensity to skin damage. CONCLUSIONS: HMSC are not immunologically reactive and can trespass species defense barriers. Animals treated with these cells repaired injuries better than controls. In this way we propose that universal HMSC from donors can be cultured, expanded, and cryopreserved to be used in human organ or tissue regeneration.

[Prevalence of anaemia in pregnancy and analysis of the underlying factors]. / Prevalencia de anemia del embarazo y análisis de sus factores condicionantes.

OBJECTIVE: To determine the prevalence of anaemia and to evaluate the factors that condition its occurrence.Setting. Out-patient clinics in the La Plata area, Buenos Aires, Argentina. DESIGN: Observational and prospective study. PARTICIPANTS: All the pregnant women consulting for the first time, excluding those with prior pathology or regular use of medical drugs. MEASUREMENTS: Anaemia was defined at values of Hb < 11 g/dl. Questionnaires were administered for general data and the type of nutrition, and a complete haematological report was compiled. RESULTS: 1218 pregnant women started the study. Anaemia was detected in 196 of them (16%), with average Hb 9.88 g/dl. Between normal and anaemic pregnant women, the following differences were found between the first and second consultations: weight (64.44 vs 59.50, p < 0.00001), family income (US$744.36 vs 568.28, p < 0.0001), kilocalories ingested (2,488.44 vs 2,204.28, p = 0.01), percentage of proteins in diet (15.73 vs 13.69, p = 0.002), and weekly iron consumption (15.24 mg vs 13.04, p < 0.0001). CONCLUSIONS: Pregnant women run a greater risk of suffering anaemia if they have diets of < 1800 kcal, < 13% proteins, less than 7 mg of iron per week, and haemic iron < 10%; and family income below US$400. Ensuring a proper diet and improving the social and economic conditions of this population group will reduce the risk of anaemia during pregnancy and its perinatal consequences.

Prevalencia de anemia del embarazo y análisis de sus factores condicionantes / Prevalence of anaemia in pregnancy and analysis of the underlying factors

Objetivo. Determinar la prevalencia de anemia y evaluar sus factores condicionantes. Emplazamiento. Consultorios externos en el área de La Plata, Buenos Aires, Argentina. Diseño. Estudio observacional, prospectivo. Participantes. Todas las gestantes que consultaron por primera vez, excluyéndose aquellas con patología previa o ingesta regular de fármacos. Mediciones . Se consideró anemia valores de Hb < 11 g/dl. Se realizaron encuestas para evaluar datos generales y tipo de alimentación y un estudio hematológico completo. Resultados . Participaron en el estudio 1.218 gestantes. Se detectó anemia en 196 de ellas (16 por ciento) siendo la Hb promedio de este grupo 9,88 g/dl. Entre aquellas embarazadas normales y anémicas se detectaron diferencias respecto al peso en primera consulta (64,44 frente a 59,50; p < 0,00001), ingreso familiar (744,36 frente a 568.28 dólares; p < 0,0001), kilocalorías ingeridas (2.488,44 frente a 2.204,28; p = 0,01), porcentaje de proteínas de la dieta (15,73 frente a 13,69; p = 0,002) y hierro semanal consumido (15,24 mg frente a 13,04; p < 0,0001).Conclusiones. En su primera consulta, un 16 por ciento de gestantes estaban anémicas. Existe mayor riesgo de presentar anemia en aquellas gestantes con dietas que contienen < 1.800 kcal, < de 13 por ciento de proteínas, valores < 7 mg de hierro semanal, hierro hémico < al 10 por ciento e ingreso mensual familiar < 400 dólares. Asegurar una dieta adecuada y mejorar las condiciones socioeconómicas de esta población disminuirá el riesgo de anemia durante la gestación y sus consecuencias perinatales (AU)

Graft versus host disease in autologous stem cell transplantation.

Relapse remains the major cause of mortality in haematological malignancies treated with autologous stem cell transplantation (ASCT). Graft versus tumour reaction (GVT) associated to autologous graft versus host disease (GVDH) may contribute to eliminate minimal residual disease (MRD) after ASCT. Eighty patients with several diagnostics were submitted to ASCT. After stem cell infusion, patients randomised in 4 groups. Groups were treated as follows: Group A received either a IFN (alpha Interferon--1,000,000 U/d), Cyclosporine A (CSA--1 mg/-kg/d intravencus) for 28 days, and granulocyte-macrophage colony stimulating factor (GM-CSF-250/m2/d) until engraftment; B: CSA (same dose and way) and GM-CSF; C: CSA (1 mg/kg/d orally) and GM-CSF and D: only GM-CSF. Patients were inspected daily and if skin rash was detected, a skin biopsy was obtained at that moment, otherwise biopsies were obtained at day 21 after ASCT. GVHD was positive in 23 patients (13 from group A and 10 from group B). All cases were grades I and II. A majority of CD4+ T lymphocytes was seen in skin infiltrates. No significant differences were seen in WBC and platelets engraftment times, antibiotic administration or hospitalisation days required among the four groups. With a median follow up of 18 months, there were no differences in disease free survival (DFS) or overall survival (OS) between the patients who developed GVHD and the others. However, considering that myeloma cells do not express antigen MCH II, which is necessary for GVT effect, we excluded patients with multiple myeloma (MM) from survival analysis, thus obtaining a significant difference in OS results between patients who developed GVHD and those in whom this reaction was not observed (81% vs 58% p:0.05). We conclude that pharmacological induction of GVHD in ASCT is possible with CSA administration (1 mg/kg/d i.v.). Development of GVHD showed a better outcome for patients in our study except for those patients with MM. This results must be confirmed by a longer follow up of our patients and further studies.