The secretome from human-derived mesenchymal stem cells augments the activity of antitumor plant extracts in vitro.

Cancer is understood as a multifactorial disease that involve multiple cell types and phenotypes in the tumor microenvironment (TME). The components of the TME can interact directly or via soluble factors (cytokines, chemokines, growth factors, extracellular vesicles, etc.). Among the cells composing the TME, mesenchymal stem cells (MSCs) appear as a population with debated properties since it has been seen that they can both promote or attenuate tumor progression. For various authors, the main mechanism of interaction of MSCs is through their secretome, the set of molecules secreted into the extracellular milieu, recruiting, and influencing the behavior of other cells in inflammatory environments where they normally reside, such as wounds and tumors. Natural products have been studied as possible cancer treatments, appealing to synergisms between the molecules in their composition; thus, extracts obtained from Petiveria alliacea (Anamu-SC) and Caesalpinia spinosa (P2Et) have been produced and studied previously on different models, showing promising results. The effect of plant extracts on the MSC secretome has been poorly studied, especially in the context of the TME. Here, we studied the effect of Anamu-SC and P2Et extracts in the human adipose-derived MSC (hAMSC)-tumor cell interaction as a TME model. We also investigated the influence of the hAMSC secretome, in combination with these natural products, on tumor cell hallmarks such as viability, clonogenicity, and migration. In addition, hAMSC gene expression and protein synthesis were evaluated for some key factors in tumor progression in the presence of the extracts by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) and Multiplex, respectively. It was found that the presence of the hAMSC secretome did not affect the cytotoxic or clonogenicity-reducing activities of the natural extracts on cancer cells, and even this secretome can inhibit the migration of these tumor cells, in addition to the fact that the profile of molecules can be modified by natural products. Overall, our findings demonstrate that hAMSC secretome participation in TME interactions can favor the antitumor activities of natural products.

The Solent Strait: Water quality trends within a heavily trafficked marine environment, 2000 to 2020.

This study presents an important long-term historical analysis of water quality in an internationally crucial waterway (the Solent, Hampshire, UK), in the context of increasing adoption of open-loop Exhaust Gas Cleaning Systems by shipping. The pollutants studied were acidification (pH), zinc, and benzo [a] pyrene, alongside temperature. We compared baseline sites to locations likely to be impacted by pollution. The Solent's average water temperature is slightly increasing, with temperatures at wastewater sites significantly higher. Acidification suggests a complex story, with a highly significant small overall increase in pH during the study period but significantly different values at wastewater and port sites. Zn concentrations have significantly reduced but increased in enclosed waters such as marinas. BaP showed no long-term trend with values at marinas significantly and consistently higher. The findings provide valuable long-term background data and insights that can feed into the upcoming review of the European Union's Marine Strategy Framework Directive and ongoing discussions about the regulation of, and future monitoring and management strategies for coastal/marine waterways.

The impact of an annual major recreational boating event on water quality in the Solent Strait.

A long-term historical analysis of the impacts of recreational boating on marine surface water quality during a regatta (Cowes Week) in an internationally crucial waterway, the Solent Strait (Hampshire, UK) is presented. Water quality indicators studied included nitrogen concentration, bacterial indicators, and oxygen saturation, at three sampling sites at/near Cowes during 2001-2019. Findings include that sewage discharge from recreational boats is the key contributor to localised faecal contamination of marine surface waters, putting bathers and shellfisheries at risk. Bathing water quality monitoring and pollution warning systems should be strengthened prior to and during this type of regatta and access to bathing water areas may need to be restricted. These findings have implications for the regulation, future monitoring and management strategies for discharges from recreational boats during extended regattas. Adequate and affordable local facilities for recovering sewage wastewater from recreational boats should be provided alongside appropriate mechanisms for communication to sailors.

Dermo-epidermal organotypic cultures for in vitro evaluation of skin irritation and corrosion.

In recent years, in-vitro skin models for chemical hazard identification have been developed. Most of them consist only of human keratinocytes, neglecting the contribution of other skin constituents. Cultures containing the dermal and epidermal component provide an attractive system to investigate, in a more realistic model, toxicological responses, which represents a distinct advantage over keratinocytes-based models that do not mimic faithfully the in vivo environment. This study aimed to validate dermo-epidermal organotypic cultures (ORGs) as a platform to perform irritation and corrosion tests. Skin models were constructed by seeding keratinocytes on fibroblast-containing fibrin gels. After 21 days, the ORGs were evaluated histologically, and the irritant and corrosion potential was determined by means of viability measurements (MTT assay) and cytokine release, according to 431 and 439 OECD tests guidelines. Skin models showed similar histological characteristics to native skin and were able to classify different substances with high accuracy, showing their applicability to skin irritation and corrosion tests. Although cytokines release seems to be chemical-dependent, a tendency was observed, leading to the improvement of the prediction capacity. Nevertheless, further studies should be done to reduce variability in order to increase prediction capacity.

[Pelvic organ prolapse during pregnancy: treatment with vaginal pessary. A report of two cases]. / Prolapso de órganos pélvicos durante el embarazo tratado con pesario. Reporte de dos casos.

Background: Pelvic organ prolapse during pregnancy is a rare condition that may be associated with maternal and fetal complications. Objetive: To describe the experience of two cases of pelvic organ prolapse during pregnancy managed with vaginal pessary. Case report: The cases of two patients with grade 4 vaginal prolapse during pregnancy managed with vaginal pessary until the time of delivery are presented. The patients had improvement of their symptoms without complications related to prolapse or pessary use. Conclusion: Early recognition, monitoring and management of this condition are essential. The vaginal pessary is a good option for transient treatment and is associated with few complications.

Carácterísticas histológicas de piel cultivada in vitro / Histological Characteristics Of Skin Culture In Vitro

Los injertos de piel cultivados in vitro han sido utilizados tanto en la regeneración de tejidos de áreas cruentas de la piel (úlceras crónicas y quemaduras de diversos grados), como para el tratamiento de genodermatosis. En nuestro medio existe un alto índice de pacientes con úlceras crónicas y un total de 2319 pacientes quemados, en un período de 10 años. El tratamiento convencional de estos pacientes genera estadías de hospitalización prolongadas y costos hospitalarios muy elevados. En este trabajo se establecieron las condiciones para el cultivo y expansión de queratinocitos y fibroblastos humanos, con el propósito de generar un equivalente cutáneo. A su vez, se evaluaron sus características histológicas con el objeto de ofrecer otras opciones de tratamiento. Las células se obtuvieron a partir de piel proveniente de donantes de órganos y de sobrantes de procedimientos quirúrgicos. Se logró un mayor éxito en la obtención de cultivos primarios, con muestras provenientes de donantes menores de 40 años (65%), comparado con los obtenidos de mayores (33%). En el equivalente cutáneo producido con estas células se demostró que los queratinocitos y los fibroblastos, presentan características funcionales, estructurales y morfológicas semejantes a la piel intacta. El equivalente cutáneo además de conservar las características funcionales y estructurales de la piel intacta, presenta otras ventajas en términos de costos, manipulación y estabilidad frente a otros productos similares importados.

In vitro skin culture have been used in the regeneration of skin wound (chronic ulcers and burns), and for genodermatosis treatment. In our country there is a high patient number with chronic ulcers and 2319 burned in a period of 10 years. Conventional treatment generates long hospitalization stays and high costs. We established culture conditions of keratinocytes and fibroblasts expansion, to generate a cutaneous substitute in order to offer other treatment options. Skin cells were obtained from organs donors and surgical surpluses procedures. Major success was achieved in primary cultures obtained from 40-year-old younger donors samples (65%), compared with older donors (33%). In one cutaneous substitute produced with these cells, was demonstrated that keratinocytes and fibroblasts, presented functional, structural and morphologic characteristics similar to normal skin. Cutaneous substitute besides preserve normal skin functional and structural characteristic, compared with other similar imported products, our cutaneous substitute, showed many advantages in terms of costs, manipulation and stability.

Modelación de funciones de crecimientoaplicadas a la producción animal / Modeling of growth functions applied to animal production

El crecimiento animal es uno de los aspectos más importantes al momento de evaluar la productividaden las explotaciones dedicadas a la producción de carne y en algunos casos es usado como criterio deselección, sin embargo, debe tenerse en cuenta que el crecimiento no se debe exclusivamente a factoresgenéticos sino también, a efectos ambientales. Para medir el crecimiento animal se han usado diferentesmodelos matemáticos lineales, no lineales y logarítmicos, entre otros, eligiéndolos por su bondad de ajustey la facilidad de interpretación biológica de sus parámetros. Recientemente se han usado modelos mixtosen los que sus parámetros están compuestos de efectos fijos y efectos aleatorios, representando los valoresesperados y la varianza de los primeros, respectivamente, lo que permite evaluar la variabilidad de lasdiferentes curvas entre los individuos de una población, así como la covarianza entre los parámetros. Loscriterios más usados para elegir la curva que mejor ajusta a los datos son: el coeficiente de determinación,el porcentaje de curvas significativas y atípicas encontradas para cada función; además se pueden aplicarcriterios como el criterio de información Akaike y el criterio de información Bayesiano. El objetivo delpresente trabajo es indicarle al lector una aplicación de los modelos no lineales y no lineales mixtos en elanálisis del crecimiento animal.

Animal growth is one of the most important aspect for evaluating animal productivity in beef cattleenterprises and in some cases it is used as a criterion of selection, nevertheless, the fact that animalgrowth is not exclusively due to genetic factors but also, to environmental effects should be keep inmind. Measurement of animal growth have been performed by several logarithmic, not lineal, lineal, andmathematical models, having as selection criteria its fitness of adjustment and the feasibility for biologicalinterpretation of their parameters. Recently the mixed models have been used in which their parametersare composed of fixed and random effects representing the expected values and variance of the fixedones, respectively, which permits to evaluate the variability of different curves between individuals of apopulation, as well as the covariance between parameters. The most used criteria for selection of the curvethat best fit data are: determination coefficient and the percent of significant and atypical curves foundfor each function. In addition, other models as the Akaike information criteria and Bayesian informationcriteria can also be applied. The objective of the present review is to provide the criteria for application oflinear and non linear models when analyzing animal growth.

Cholesterol level and symptomatic hemorrhagic transformation after ischemic stroke thrombolysis.

BACKGROUND: Prestroke statin use may improve ischemic stroke outcomes, yet there is also evidence that statins and extremely low cholesterol levels may increase the risk of intracranial hemorrhage. We evaluated the independent effect of statin use and admission cholesterol level on risk of symptomatic hemorrhagic transformation (sHT) after recanalization therapy for acute ischemic stroke. METHODS: We analyzed ischemic stroke patients recorded in a prospectively maintained registry that received recanalization therapies (IV or intra-arterial fibrinolysis or endovascular embolectomy) at a university medical center from September 2002 to May 2006. The independent effect of premorbid statin use on sHT post intervention was evaluated by logistic regression, adjusting for prognostic and treatment variables known to predict increased HT risk after ischemic stroke. RESULTS: Among 104 patients, mean age was 70 years, and 49% were men. Male sex, hypertension, statin use, low total cholesterol and low-density lipoprotein (LDL) cholesterol, current smoking, elevated glucose levels, and higher admission NIH Stroke Scale (NIHSS) score were all associated with a greater risk of sHT in univariate analysis. After adjusting for covariates, low LDL cholesterol (odds ratio [OR], 0.968 per 1-mg/dL increase; 95% CI, 0.941 to 0.995), current smoking (OR, 14.568; 95% CI, 1.590 to 133.493), and higher NIHSS score (OR, 1.265 per 1-point increase; 95% CI, 1.047 to 1.529) were independently associated with sHT risk. CONCLUSIONS: Lower admission low-density lipoprotein cholesterol level with or without statin use, current smoking, and greater stroke severity are associated with greater risk for symptomatic hemorrhagic transformation after recanalization therapy for ischemic stroke.

Efecto de la pluviosidad y el brillo solar sobre la producción y características del semen en el pez Brycon henni (Pisces: Characidae) / Effect of pluviosity and sun shine on sperm production and seminal characteristics of the fish Brycon henni (Pisces: Characidae)

In Colombia the fish Brycon henni is a protected endemic species. It inhabits water bodies in coffee producing areas (700-1900 ma.s.l.; 4º35’56’’ N -74º04’51’’ W; 18-28°C). Insufficient knowledge of its basic biology and behavior prevent the commercial culture of this promising fish. We studied the production and sperm physiology of captive males. Along a year 20 samples were taken from each of 10 males. The sample was obtained by abdominal cefalo-caudal massage and transported to the laboratory at 4°C. Except for September and October (maximum rainfall),sperm was always obtained in at least 50% of the males. Color, osmolality and pH were similar in all the samples. Volume, concentration, viability, motility and activation time were variable: sunshine had a positive effect on volume (Spearman p<0.05) and on sperm concentration (Spearman p<0.05) while pluviosity had a negative effect on volume and viability. The proportion of ions (Na,K,Mg,Ca)was constant along the year; Na being 10-fold higher than K and 100-fold higher than Mg and Ca; however the absolute concentration of all ions was slightly higher in April and in July (with no apparent relation with the other variables analyzed).

El pez Brycon henni es una especie endémica protegida por la legislación colombiana, que habita cuerpos de agua de zonas cafeteras (700-1900 m. s. n. m),comprendidas entre los 4º35’56’’ N y 74º04’51’’ W, con temperaturas que oscilan entre los 18 y los 28ºC. A pesar de las características promisorias de esta especie, su reproducción en cautiverio a nivel comercial no ha sido posible por falta de conocimientos básicos de su biología y comportamiento. El objetivo del presente trabajo fue caracterizar la producción y la fisiología espermática de machos en cautiverio. A lo largo de un año se tomaron 20 muestras de cada uno de 10 ejemplares. El semen se obtuvo mediante masaje abdominal cráneo-caudal y se transportó a 4ºC para su análisis en el laboratorio. Con excepción de septiembre y octubre que fueron los meses más lluviosos, siempre se obtuvo semen de al menos el 50% de los animales. El color, la osmolalidad y el pH fueron similares en todas las muestras a lo largo del año. El volumen, la concentración, la viabilidad, la movilidad y el tiempo de activación fueron variables: El efecto del brillo solar fue positivo sobre el volumen (Spearman p<0.05) y sobre la concentración espermática (Spearman p<0.05), mientras que la pluviosidad tuvo un efecto negativo sobre el volumen y la viabilidad. La proporción entre los iones (Na, K, Mg, Ca) se mantuvo constante durante todo el año: el sodio fue el mayor, seguido por el potasio, y por el magnesio y el calcio, 10 y 100 veces menos, respectivamente; sin embargo la concentración absoluta de estos iones aumentó durante los meses de abril y julio sin una relación aparente con las otras variables analizadas.

Mitochondrial activity, distribution and segregation in bovine oocytes and in embryos produced in vitro.

Contents Bovine oocytes and embryos produced in vitro were studied to determine the mitochondrial pattern of distribution, segregation and activity using DIOC 6 and Jc-1 fluorescence. The highest fluorescence level observed in mature oocytes was taken as 100% activity and six activity levels were estimated as follows: (1) 0%, (2) 1-15%, (3) 16-30%, (4) 31-50%, (5) 51-75% and (6) 76-100%. Three patterns of mitochondrial distribution were found: (1) diffused throughout the cytoplasm in oocytes and embryos, (2) pericytoplasmic in oocytes and embryos, and (3) perinuclear only in embryos. The segregation of mitochondria in blastomeres showed two distinct patterns: (1) symmetrical with an even mitochondrial population, and (2) asymmetrical with different numbers of mitochondria in each blastomere. In immature oocytes, mitochondrial activity was very low and the distribution was diffuse or negligible, while in mature oocytes the activity was high and the distribution was diffuse or pericytoplasmic. Competent embryos up to the 16-cell stage showed intermediate levels of activity (16-50%) but activity decreased thereafter up to the blastocyst stage. Non-competent embryos showed low levels of activity (1-15%) at all stages. These results suggest that mitochondria might play an important role during early development and that a minimum threshold of activity regulates the potential competence for reaching the blastocyst stage.

[Effect of pluviosity and sun shine on sperm production and seminal characteristics of the fish Brycon henni (Pisces: Characidae)]. / Efecto de la pluviosidad y el brillo solar sobre la producción y características del semen en el pez Brycon henni (Pisces: Characidae).

Effect of pluviosity and sun shine on sperm production and seminal characteristics of the fish Brycon henni (Pisces: Characidae). In Colombia the fish Brycon henni is a protected endemic species. It inhabits water bodies in coffee producing areas (700-1900 ma.s.l.; 4 degrees 35'56" N - 74 degrees 04'51" W; 18-28 degrees C). Insufficient knowledge of its basic biology and behavior prevent the commercial culture of this promising fish. We studied the production and sperm physiology of captive males. Along a year 20 samples were taken from each of 10 males. The sample was obtained by abdominal cefalo-caudal massage and transported to the laboratory at 4 degrees C. Except for September and October (maximum rainfall), sperm was always obtained in at least 50% of the males. Color, osmolality and pH were similar in all the samples. Volume, concentration, viability, motility and activation time were variable: sunshine had a positive effect on volume (Spearman p < 0.05) and on sperm concentration (Spearman p < 0.05) while pluviosity had a negative effect on volume and viability. The proportion of ions (Na, K, Mg, Ca) was constant along the year; Na being 10-fold higher than K and 100-fold higher than Mg and Ca; however the absolute concentration of all ions was slightly higher in April and in July (with no apparent relation with the other variables analyzed).

Human skin keratinocytes modified by a Friend-derived retroviral vector: a functional approach.

The goal of this study was to test the efficiency and possible functional effects of a Friend Leukemia derived retrovirus vector (FOCH29-NeoR) on cultured human keratinocytes, obtained from skin biopsy samples. The keratinocytes were grown and infected with filtered Friend vector supernatant. After one or two doses of infection, one duplicate of the culture was submitted to selection with G418; the other one was utilized for DNA extraction and PCR modification detection. Transduction efficiency was 46.66 percent and 47.22 percent for one and two doses of infection respectively (range 100 to 15 %). Colony Forming Efficiency (CFE) assays were done with Rodhamine-B staining in nonselected modified cultures and negative controls. There was no difference in CFE (% CFE= 10.74+/-6.53 negative control vs % CFE= 9.22+/-5.45 with one dose, and % CFE= 10.03+/-5.74 with two doses of infection). Nevertheless, the cell-cycle analysis done by Propidium Iodade (PI) incorporation and colchicine-arrest assays in nonselected transduced and nontransduced cells show that transduced keratinocytes have a longer time to enter G2. As far as we know, this is the first report of retroviral transduction-induced changes in the cell cycle done on human keratinocytes. This observation is very important because retroviral vectors of genes, such as platelet derived growth factor (PDGF) or vascular endothelial growth factor (VEGF), are expected to facilitate the implementation of these modified cultures for tissue grafting and skin substitute development and potentiate the effectiveness of the grafts.

Aggregation of sphalerite: role of zinc ions.

Sphalerite from six sources is shown to aggregate at ca. pH 7-9, confirmed by different techniques (settling velocity, suspension analysis, and optical microscopy). This does not correlate with the isoelectric point, which is consistently < pH 6. A similar observation was made by Healy and Jellet [J. Colloid Interface Sci. 24 (1967) 41-46] for ZnO (and reconfirmed here), who suggested Zn hydrolysis products were responsible. The hypothesis is tested using chalcopyrite and silica suspensions in the presence of Zn ions. It is found that aggregation occurs over the same pH range, 7-9. Solution and surface analysis (extraction by EDTA) shows sphalerite released sufficient Zn ions to promote aggregation. The pH range 7-9 corresponds to hydroxide formation, suggesting this is the species responsible. Aggregation of sphalerite due to the polymerization/flocculation action of zinc hydroxide is proposed.

Intragraft cytokine expression in heart transplants with mild or no histological rejection.

UNLABELLED: The study of pro-inflammatory cytokines produced in situ in heart allografts may help to understand the mechanisms of rejection and open new possibilities to control graft rejection. METHODS: A total of 23 endomyocardial biopsies obtained from 16 transplanted patients treated with triple-drug therapy (azathioprine, prednisone, and cyclosporine) were studied. mRNA expression for tumor necrosis factor (TNF)-alpha, interleukin (IL)-1 beta, IL-6, IL-10, IL-12, IL-15, transforming growth factor (TGF)-beta, and beta-actin was determined by reverse transcription polymerase chain reaction (RT-PCR) and Southern blotting. Semiquantitative analysis was done by establishing the ratio between densitometric integrated value of each cytokine with the beta-actin and correlated with the histopathologic findings. RESULTS: Three groups of biopsies were determined according to the International Society for Heart and Lung Transplantation criteria: grade 0 (control group, n=12), grade 1A (sub-clinical rejection, n=6) and 'quilty effect' (n=5). An increased expression of mRNA for TNF-alpha and IL-6 (p=0.0091 and 0.0075, respectively) was found associated with rejection grade 1A episodes, mRNA for IL-1 beta was nonspecifically expressed in all the study groups, while IL-10 mRNA was not detected in any of the biopsies studied. mRNA for IL-12 and IL-15 was not associated with rejection. Interestingly, TGF-beta was not detected in any of the biopsies with the 'quilty pattern'. CONCLUSION: The association of TNF-alpha and IL-6 mRNA in situ expression with mild histologically probed rejection episodes may be used in the monitoring of heart transplants.

Reversible tacrolimus-induced neurotoxicity isolated to the brain stem.

Diplopia, nystagmus, visual hallucinations, and internuclear ophthalmoplegia developed in a 30-year-old woman 84 days after she received a matched, unrelated bone marrow transplant for chronic myeloid leukemia. A regimen of tacrolimus had been administered since the transplantation was performed. MR imaging revealed bilaterally symmetric regions of signal abnormality with abnormal contrast enhancement in the brain stem. No supratentorial abnormality was present. Tacrolimus therapy was discontinued, and the symptoms resolved. MR imaging that was performed 10 days after tacrolimus was discontinued showed resolution of the abnormalities.

Efficient transduction of hemopoietic CD34+ progenitors of human origin using an original retroviral vector derived from Fr-MuLV-FB29: in vitro assessment.

A novel retroviral vector has been designed based on a Friend-murine leukemia virus (Fr-MuLV) FB29 strain. The latter has been selected according to characteristics of pathogenicity in mice where it induces a disease of the haemopoietic system affecting all lineages. Higher infectivity has also been demonstrated as compared to other strains. In accordance with these findings, the amphotropic producer clone used in this study carrying along the neomycine resistance gene (FOCH-Neo), harbors viral titers over 10(7) cfu/ml. To investigate the potential of genetically engineering hematopoietic precursors, CD34+ progenitors were selected from cord blood, bone marrow, and peripheral blood mobilized stem cells (patients + solid tumors) and transduced with FOCH-Neo. High transduction rates were achieved using virus supernatant and minimal doses of hematopoietic growth factors during pretransduction and transduction steps. A polymerase chain reaction (PCR) assay investigating the presence of both neomycin-encoding and viral vector sequences tested positive in 45-90% of granulocyte-macrophage colony-forming units (CFU-GM) generating cells (bone marrow and peripheral blood derived cells) following transduction. An average of 35% colonies showed resistance to G418. Such levels of transduction proved reproducible using only supernatants harboring over 10(7) cfu/ml. In those experiments where long-term in vitro cultures could be maintained over 5 weeks (all cord blood and 5 among 23 PBSC), efficient transduction of long-term culture initiating cell (LTC-IC) hematopoietic progenitors was demonstrated on the basis of both resistance to G418 and virus integration. In the latter case, the PCR assay tested positive in as much as 35-60% of late unselected CFU-colonies. This novel retroviral vector harbors interesting features toward genetic modification of hematopoietic progenitors.