Case report: Neuronal intranuclear inclusion disease presenting with acute encephalopathy.

Neuronal intranuclear inclusion disease (NIID), a neurodegenerative disease previously thought to be rare, is increasingly recognized despite heterogeneous clinical presentations. NIID is pathologically characterized by ubiquitin and p-62 positive intranuclear eosinophilic inclusions that affect multiple organ systems, including the brain, skin, and other tissues. Although the diagnosis of NIID is challenging due to phenotypic heterogeneity, a greater understanding of the clinical and imaging presentations can improve accurate and early diagnosis. Here, we present three cases of pathologically proven adult-onset NIID, all presenting with episodes of acute encephalopathy with protracted workups and lengthy time between symptom onset and diagnosis. Case 1 highlights challenges in the diagnosis of NIID when MRI does not reveal classic abnormalities and provides a striking example of hyperperfusion in the setting of acute encephalopathy, as well as unique pathology with neuronal central chromatolysis, which has not been previously described. Case 2 highlights the progression of MRI findings associated with multiple NIID-related encephalopathic episodes over an extended time period, as well as the utility of skin biopsy for antemortem diagnosis.

Morphological adaptations in response to chronic exercise across musculoskeletal tissues: a systematic review / Adaptaciones morfológicas en respuesta al ejercicio crónico en los tejidos osteomusculares: una revisión sistemática

León, F., Mestre. A., Priego, L., & Vera, J.C. (2023). Morphological adaptations in response to chronic exercise across musculoskeletal tissues: a systematic review. PENSAR EN MOVIMIENTO: Revista de Ciencias del Ejercicio y la Salud, 21(1), 1-28. To date, there is no systematic review that summarizes the morphological adaptations of the musculoskeletal system in response to chronic exercise. This systematic review selected original articles published in English between 2000 and 2020, with a clear exercise intervention and presenting a morphological change in the tissue under study, and covering human participants irrespective of age, gender or health condition. In total, 2819 records were identified. After removal of duplicates, title and abstract screening and full-text review, 67 records were included in the final analysis (6 for inter-vertebral disc, 6 for cartilage, 36 for bone, 2 for ligament, 9 for tendon and 7 for muscle). The most used interventions were aerobic, resistance, and plyometric exercise. Population ranged from children and healthy active people to individuals with a health condition. In conclusion, as a response to chronic exercise there are morphological adaptations in the tissues of the musculoskeletal system which vary from increased stiffness to an increase in cross-sectional area. Although tissues can adapt, several questions still linger, such as optimal dose and type of exercise, whether adaptations can occur in an injured tissue, and functional implications of these adaptations. Future research should address these questions.

León, F., Mestre. A., Priego, L. y Vera, J.C. (2023). Adaptaciones morfológicas en respuesta al ejercicio crónico en los tejidos osteomusculares: una revisión sistemática. PENSAR EN MOVIMIENTO: Revista de Ciencias del Ejercicio y la Salud, 21(1), 1-28. Hasta la fecha, no existe una revisión sistemática que resuma las adaptaciones morfológicas del sistema osteomuscular en respuesta al ejercicio crónico. Esta revisión sistemática seleccionó artículos originales, con fecha de publicación de 2000 a 2020, idioma de publicación en inglés, con una clara intervención de ejercicio y que presentaron un cambio morfológico en el tejido estudiado. Participantes humanos independientemente de la edad, el género o condición de salud. Se identificaron 2819 registros. Después de eliminar los duplicados, la selección de títulos y resúmenes y la revisión de texto completo, se incluyeron 67 registros en el análisis final (6 para disco intervertebral, 6 para cartílago, 36 para hueso, 2 para ligamento, 9 para tendón y 7 para músculo). Los resultados destacan que las intervenciones más utilizadas fueron ejercicio aeróbico, contra resistencia y pliométrico. La población abarcó desde niños y personas sanas activas hasta personas con alguna condición de salud. Se concluye que como respuesta al ejercicio crónico existen adaptaciones morfológicas en los tejidos del sistema musculoesquelético, que pueden variar desde un aumento de rigidez hasta un aumento de área. Aunque los tejidos pueden adaptarse, aún quedan varias preguntas, como la dosis y tipo de ejercicio óptimo, si pueden ocurrir adaptaciones en un tejido lesionado y las implicaciones funcionales de estas adaptaciones. La investigación futura debe abordar estas preguntas.

León, F., Mestre. A., Priego, L. e Vera, J.C. (2023). Adaptações morfológicas em resposta ao exercício crônico nos tecidos osteomusculares: uma revisão sistemática. PENSAR EN MOVIMIENTO: Revista de Ciencias del Ejercicio y la Salud, 21(1), 1-28. Até o momento, não há uma revisão sistemática que resuma as adaptações morfológicas do sistema osteomuscular em resposta ao exercício crônico. Esta revisão sistemática selecionou artigos originais, com data de publicação de 2000 a 2020, idioma de publicação em inglês, com clara intervenção de exercícios e que apresentaram alteração morfológica no tecido estudado. Participantes humanos, independentemente da idade, sexo ou condição de saúde. Foram identificados 2.819 registros. Após eliminar os artigos duplicados, triagem de título e resumo e revisão do texto completo, 67 registros foram incluídos na análise final (6 para disco intervertebral, 6 para cartilagem, 36 para osso, 2 para ligamento, 9 para tendão e 7 para músculo). Os resultados destacam que as intervenções mais utilizadas foram exercícios aeróbicos, resistidos e pliométricos. A população variou de crianças e pessoas saudáveis ativas a pessoas com alguma condição de saúde. Conclui-se que, em resposta ao exercício crônico, ocorrem adaptações morfológicas nos tecidos do sistema musculoesquelético, que podem variar desde um aumento de rigidez até um aumento de área. Embora os tecidos possam se adaptar, várias questões permanecem, como a dose ideal e o tipo de exercício, se adaptações podem ocorrer no tecido lesado e as implicações funcionais dessas adaptações. Pesquisas futuras devem abordar essas questões.

Pain Relief After Allogenic Stem Cell Disc Therapy.

BACKGROUND: Treatment of intermediate-stage painful degenerative disc disease is controversial, with few reliable options. Allogenic mesenchymal stem cells (MSCs)are an alternative to autologous stem cell transplantation. Allogeneic MSCs in the treatment of discogenic low back pain have some practical advantages, ranging from availability to ease of treatment in a procedure-room setting. OBJECTIVES: To assess the efficacy and safety of allogenic MSC injection into painful lumbar intervertebral discs and associated clinical outcomes. STUDY DESIGN: Retrospective observational cohort study. SETTING: Private practice. METHODS: There were 33 patients: 15 women and 18 men with an average age of 47.6 years. The patients' average follow-up was 26.88 months Patients were treated with intradiscal injection of approximately 5 million allogeneic polyclonal MSCs in 1% hyaluronic acid derived from immunoselected umbilical cord stem cells. Patients were monitored for adverse event reactions. Clinical outcomes were assessed with reductions in the reported Visual Analog Scale (VAS) for back pain, the Oswestry Disability Index (ODI) scores, and the use of the modified Macnab criteria. RESULTS: No patient required any additional treatments for low back pain stemming from the level treated with MSC injections. At a 2-year follow-up, the average VAS low back score reduction was 6.565 ± 1.619 and 38.333 ± 14.865 for the ODI (P < 0.001). Reported Macnab outcomes were excellent in 11 patients (33.3%), good in 19 (57.6%), and fair in 3 (9.1%). LIMITATIONS: Our observational study is limited by patient selection, hindsight bias, and low patient numbers. CONCLUSION: The results of our feasibility study suggest that the injection of allogeneic MSCs to treat patients with painful intermediate-stage degenerative disc disease has merit. No adverse reactions were observed. The authors recommend further study in a randomized prospective study setting with a placebo control group or a natural history study group of patients to solidify this research.

Effect of Chitosan and Naringin on Enteric Methane Emissions in Crossbred Heifers Fed Tropical Grass.

In order to meet consumer needs, the livestock industry is increasingly seeking natural feed additives with the ability to improve the efficiency of nutrient utilization, alternatives to antibiotics, and mitigate methane emissions in ruminants. Chitosan (CHI) is a polysaccharide with antimicrobial capability against protozoa and Gram-positive and -negative bacteria, fungi, and yeasts while naringin (NA) is a flavonoid with antimicrobial and antioxidant properties. First, an in vitro gas production experiment was performed adding 0, 1.5, 3.0 g/kg of CHI and NA under a completely randomized design. The substrate containing forage and concentrate in a 70:30 ratio on a dry matter (DM) basis. Compounds increased the concentration of propionic acid, and a significant reduction in methane production was observed with the inclusion of CHI at 1.5 g/kg in in vitro experiments (p < 0.001). In a dry matter rumen degradability study for 96 h, there were no differences in potential and effective degradability. In the in vivo study, six crossbred heifers fitted with rumen cannulas were assigned to a 6 × 6 Latin square design according to the following treatments: control (CTL), no additive; chitosan (CHI1, 1.5 g/kg DMI); (CHI2, 3.0 g/kg DMI); naringin (NA1, 1.5 g/kg DMI); (NA2, 3.0 g/kg DMI) and a mixture of CHI and NA (1.5 + 1.5 g/kg DMI) given directly through the rumen cannula. Additives did not affect rumen fermentation (p > 0.05), DM intake and digestibility of (p > 0.05), and enteric methane emissions (p > 0.05). CHI at a concentration of 1.5 g/kg DM in in vitro experiments had a positive effect on fermentation pattern increasing propionate and reduced methane production. In contrast, in the in vivo studies, there was not a positive effect on rumen fermentation, nor in enteric methane production in crossbred heifers fed a basal ration of tropical grass.

Caracterización clínica y metabólica de pacientes con diagnóstico de urolitiasis atendidos en una clínica de cuarto nivel de Barranquilla, Colombia / Clinical and metabolic characterization of patients with a diagnosis of urolithiasis in a four-level clinic in the city of Barranquilla

Resumen Introducción: la urolitiasis es una enfermedad frecuente de la cual en Colombia se han publicado estudios previos; sin embargo, estos no comparan las características sociodemográficas y clínicas de los pacientes con las comorbilidades y los factores predisponentes de litiasis como hiperuricemia, hipertensión arterial (HTA), obesidad y enfermedad renal crónica (ERC). Objetivos: caracterizar clínica y metabólicamente los pacientes con diagnóstico de urolitiasis atendidos en una clínica de cuarto nivel de Barranquilla, Colombia, en el año 2019. Materiales y métodos: se realizó un estudio observacional, descriptivo y transversal en 49 pacientes con base en el estudio de fichas clínicas Resultados: el 53,1 % de los participantes eran hombres y las medianas de edad y de índice de masa corporal (IMC) fueron 58 años y 26,4 kg/m2, respectivamente. Algunas de las comorbilidades identificadas fueron, en orden de frecuencia, HTA (69,4 %), ERC (36,7 %), infección de vías urinarias recurrente (24,5 %), hiperuricemia (44,9 %), hipercalcemia (16,3 %) e hiperfosfatemia (12,2 %). Los tipos de cristal encontrados fueron oxalato (20,4 %), urato (12,2 %), mezcla de oxalato y urato (4,1 %), fosfato (4,1 %), hipercalciuria e hiperoxaluria (38,8 %), hiperuricosuria e hipocalciuria (18,4 %) y hipofosfaturia o hipofosfaturia (4,1 %). Asimismo, la hiperuricemia se asoció a edad (p=0,028), ERC (p=0,026), medicamentos antihipertensivos (p=0,022), posición del cálculo en cáliz renal (p=0,012), hiperparatiroidismo (p=0,007), depuración de creatinina (p=0,046) e hipercalciuria (p=0,049). El IMC ≥30 se asoció con ERC estadio 5 (p=0,025), diálisis (p=0,025) e hiperoxaluria (p=0,021). Conclusiones: en la población analizada se evidenció una frecuencia significativa de ERC, hiperuricemia, obesidad e HTA.

Abstract Introduction: Urolithiasis is a disease with high frequency and our environment is no exception. Previous studies have been published in Colombia, however, these do not compare the sociodemographic and clinical characteristics of patients with comorbidities and predisposing factors for lithiasis such as they are hyperuricemia, high blood pressure, obesity, and chronic kidney disease (CKD). Objectives: To characterize clinically and metabolically the patients diagnosed with urolithiasis in a fourth-level clinic in the city of Barranquilla in 2019. Methods: Observational, descriptive, cross-sectional study. In 49 patients, based on the study of clinical records Results: The median age was 58 years, the male sex in 53.1 %. The median body mass index was 26.4 Kg / m2. High blood pressure was identified in 69.4 %, chronic kidney disease (CKD) in 36.7 %, recurrent urinary tract infection in 24.5 %. Hyperuricemia in 44.9 %, hypercalcemia in 16.3 % and hyperphosphatemia in 12.2 %. The crystal types were oxalate in 20.4 %, urate in 12.2 %, mixture of the previous ones in 4.1 % and in the same proportion phosphate. Hypercalciuria and hyperoxaluria in 38.8 %, hyperuricosuria and hypocalciuria in 18.4 %, while hyperphosphaturia or hypophosphaturia in 4.1 %. Hyperuricemia was associated with age (p = 0.028), CKD (p = 0.026), antihypertensive drugs (p = 0.022), the position of the stone in the renal calyx (p = 0.012), hyperparathyroidism (p = 0.007), creatinine clearance (p = 0.046) and hypercalciuria (p = 0.049). BMI ≥30 was associated with stage 5 CKD (p = 0.025), dialysis (p = 0.025), and hyperoxaluria (p = 0.021). Conclusions: A significant frequency of CKD, hyperuricemia, obesity and hypertension was evidenced in patients with urolithiasis.

Role of Secondary Plant Metabolites on Enteric Methane Mitigation in Ruminants.

The rumen microbiome plays a fundamental role in all ruminant species, it is involved in health, nutrient utilization, detoxification, and methane emissions. Methane is a greenhouse gas which is eructated in large volumes by ruminants grazing extensive grasslands in the tropical regions of the world. Enteric methane is the largest contributor to the emissions of greenhouse gases originating from animal agriculture. A large variety of plants containing secondary metabolites [essential oils (terpenoids), tannins, saponins, and flavonoids] have been evaluated as cattle feedstuffs and changes in volatile fatty acid proportions and methane synthesis in the rumen have been assessed. Alterations to the rumen microbiome may lead to changes in diversity, composition, and structure of the methanogen community. Legumes containing condensed tannins such as Leucaena leucocephala have shown a good methane mitigating effect when fed at levels of up to 30-35% of ration dry matter in cattle as a result of the effect of condensed tannins on rumen bacteria and methanogens. It has been shown that saponins disrupt the membrane of rumen protozoa, thus decreasing the numbers of both protozoa and methanogenic archaea. Trials carried out with cattle housed in respiration chambers have demonstrated the enteric methane mitigation effect in cattle and sheep of tropical legumes such as Enterolobium cyclocarpum and Samanea saman which contain saponins. Essential oils are volatile constituents of terpenoid or non-terpenoid origin which impair energy metabolism of archaea and have shown reductions of up to 26% in enteric methane emissions in ruminants. There is emerging evidence showing the potential of flavonoids as methane mitigating compounds, but more work is required in vivo to confirm preliminary findings. From the information hereby presented, it is clear that plant secondary metabolites can be a rational approach to modulate the rumen microbiome and modify its function, some species of rumen microbes improve protein and fiber degradation and reduce feed energy loss as methane in ruminants fed tropical plant species.

Influence of energy supplementation on dietary nitrogen utilization and milk production in cows fed foliage of Leucaena leucocephala.

The aim of the study was to assess the effect of four energy supplements (two highly fermentable; two starch-based carbohydrates) on blood urea nitrogen (BUN), urinary urea excretion, and milk yield, in dual-purpose cows fed foliage of Leucaena leucocephala (Leucaena). Five Holstein-Zebu cows with 450 kg body weight in their second third of lactation were used in a 5 × 5 Latin square design. Cows were fed (dry basis) a mixture of 45% Leucaena and 55% Pennisetum purpureum grass. Treatments were supplementation with (i) sugarcane molasses (Mo), (ii) sorghum grain (So), (iii) fresh citrus pulp (CitP) or (iv) rice polishing (RP), all of them incorporated into the diet at 25 MJ of ME/cow/day. There was a control group (Cont) without energy supplementation. The study comprised five periods of 20 days (15 days adaptation, 5 days measurements). Dry matter intake (kg/day) was lower (P < 0.05) for Cont (9.4) compared with Mo (12.1), So (12.0), CitP (11.9) and RP (11.9) but no difference was observed among energy supplements (P > 0.05). Milk yield (kg/day) was higher (P < 0.05) in cows supplemented with starch supplements (4.7 for So; 4.9 for RP) compared with Cont (3.3). Milk yield from highly fermentable supplements (Mo and CitP) did not differ (P > 0.05) from Cont or other treatments. Milk protein, fat and lactose were not different among treatments (P > 0.05). Blood urea nitrogen and urinary urea excretion were both reduced (P < 0.05) by energy supplementation. Urinary urea excretion was not different (P > 0.05) among cows fed different sources of energy. It is concluded that in dual-purpose cows fed Leucaena foliage, supplementation with sugarcane molasses, citrus pulp or rice polishing reduced blood urea nitrogen and urinary urea excretion. Milk yield was increased by sorghum and rice polishing whereas energy supplementation did not affect milk composition.

Effect of Dried Leaves of Leucaena leucocephala on Rumen Fermentation, Rumen Microbial Population, and Enteric Methane Production in Crossbred Heifers.

The effects of dietary inclusion of dried Leucaena leucocephala leaves (DLL) on nutrient digestibility, fermentation parameters, microbial rumen population, and production of enteric methane (CH4) in crossbred heifers were evaluated. Four heifers were used in a 4 × 4 Latin square design consisting of four periods and four levels of inclusion of DLL: 0%, 12%, 24%, and 36% of dry matter (DM) intake. Results showed that DM intake (DMI), organic matter intake, and gross energy intake (GEI) were similar (p > 0.05) among treatments. Apparent digestibility of organic matter, neutral detergent fiber, and energy decreased with increasing levels of DLL in the ration (p < 0.05). In contrast, digestible crude protein (CP) was higher (p < 0.05) in treatments with 12% and 24% DM of DLL. The inclusion of DLL did not affect (p > 0.05) rumen pH and total volatile fatty acids. Rumen microbial community was not affected (p > 0.05) by treatment. There was a linear reduction (p < 0.05) in CH4 emissions as the levels of DLL in the ration were increased. Results of this study suggest that an inclusion of 12% DM of ration as DLL enhances digestible CP and reduces daily production of enteric CH4 without adversely affecting DMI, rumen microbial population, and fermentation parameters.

Construction and Operation of a Respiration Chamber of the Head-Box Type for Methane Measurement from Cattle.

This paper aims to describe the construction and operation of a respiration chamber of the head-box type for methane (CH4) measurements in bovines. The system consists of (1) a head box with a stainless steel frame and acrylic walls, floor, and ceiling; (2) a stainless steel feeder; (3) an automatic drinking water bowl; (4) a hood made from reinforced canvas; (5) an infrared (IR) CH4 gas analyzer, a mass flow generator, a data-acquisition system; and (6) a steel metabolic box. Six assays were conducted to determine the pure CH4 recovery rate of the whole system in order to validate it and comply with standards of chamber operation. The gravimetrical method was used for the recovery test and the recovery rate obtained was 1.04 ± 0.05. Once the system was calibrated, measurements of CH4 were conducted using eight animals consisting of four Holstein cows with a live weight of 593.8 ± 51 kg and an average milk yield of 23.3 ± 1.8 kg d-1 and four heifers with a live weight of 339 ± 28 kg. The CH4 production values were 687 ± 123 and 248 ± 40 L CH4 d-1 for cows and heifers, respectively. The CH4 yield was 19.7 ± 3.4 g and 17.1 ± 3.4 g CH4 kg-1 of dry matter consumed for cows and heifers, respectively. These results are consistent with those reported in the literature.

The Role of Chitosan as a Possible Agent for Enteric Methane Mitigation in Ruminants.

Livestock production is a main source of anthropogenic greenhouse gases (GHG). The main gases are CH4 with a global warming potential (GWP) 25 times and nitrous oxide (N2O) with a GWP 298 times, that of carbon dioxide (CO2) arising from enteric fermentation or from manure management, respectively. In fact, CH4 is the second most important GHG emitted globally. This current scenario has increased the concerns about global warming and encouraged the development of intensive research on different natural compounds to be used as feed additives in ruminant rations and modify the rumen ecosystem, fermentation pattern, and mitigate enteric CH4. The compounds most studied are the secondary metabolites of plants, which include a vast array of chemical substances like polyphenols and saponins that are present in plant tissues of different species, but the results are not consistent, and the extraction cost has constrained their utilization in practical animal feeding. Other new compounds of interest include polysaccharide biopolymers such as chitosan, mainly obtained as a marine co-product. As with other compounds, the effect of chitosan on the rumen microbial population depends on the source, purity, dose, process of extraction, and storage. In addition, it is important to identify compounds without adverse effects on rumen fermentation. The present review is aimed at providing information about chitosan for dietary manipulation to be considered for future studies to mitigate enteric methane and reduce the environmental impact of GHGs arising from livestock production systems. Chitosan is a promising agent with methane mitigating effects, but further research is required with in vivo models to establish effective daily doses without any detrimental effect to the animal and consider its addition in practical rations as well as the economic cost of methane mitigation.

Data on SVCT2 transporter expression and localization in cancer cell lines and tissues.

The data presented in this article are related to the research paper entitled "Increased expression of mitochondrial sodium-coupled ascorbic acid transporter-2 (mitSVCT2) as a central feature in breast cancer", available in Free Radical Biology and Medicine Journal [1]. In this article, we examined the SVCT2 transporter expression in various breast cancer cell lines using RT-PCR and Western blot assays. In addition, we analyzed the subcellular localization of SVCT2 by immunofluorescence colocalization assays and cellular fractionation experiments. Finally, an analysis of different cancer tissue microarrays immunostained for SVCT2 and imaged by The Human Protein Atlas (https://www.proteinatlas.org) is presented.

GLUT1 and GLUT8 support lactose synthesis in Golgi of murine mammary epithelial cells.

The mammary gland increases energy requirements during pregnancy and lactation to support epithelial proliferation and milk nutrients synthesis. Lactose, the principal carbohydrate of the milk, is synthetized in the Golgi of mammary epithelial cells by lactose synthase from glucose and UPD galactose. We studied the temporal changes in the expression of GLUT1 and GLUT8 in mammary gland and their association with lactose synthesis and proliferation in BALB/c mice. Six groups were used: virgin, pregnant at 2 and 17 days, lactating at 2 and 10 days, and weaning at 2 days. Temporal expression of GLUT1 and GLUT8 transporters by qPCR, western blot and immunohistochemistry, and its association with lactalbumin, Ki67, and cytokeratin 18 within mammary tissue was studied, along with subcellular localization. GLUT1 and GLUT8 transporters increased their expression during mammary gland progression, reaching 20-fold increasing in GLUT1 mRNA at lactation (p < 0.05) and 2-fold at protein level for GLUT1 and GLUT8 (p < 0.05 and 0.01, respectively). The temporal expression pattern was shared with cytokeratin 18 and Ki67 (p < 0.01). Endogenous GLUT8 partially co-localized with 58 K protein and α-lactalbumin in mammary tissue and with Golgi membrane-associated protein 130 in isolated epithelial cells. The spatial-temporal synchrony between expression of GLUT8/GLUT1 and alveolar cell proliferation, and its localization in cis-Golgi associated to lactose synthase complex, suggest that both transporters are involved in glucose uptake into this organelle, supporting lactose synthesis.

Increased expression of mitochondrial sodium-coupled ascorbic acid transporter-2 (mitSVCT2) as a central feature in breast cancer.

The potential role of vitamin C in cancer prevention and treatment remains controversial. While normal human cells obtain vitamin C as ascorbic acid, the prevalent form of vitamin C in vivo, the uptake mechanisms by which cancer cells acquire vitamin C has remained unclear. The aim of this study is to characterize how breast cancer cells acquire vitamin C. For this, we determined the expression of vitamin C transporters in normal and breast cancer tissue samples, and in ZR-75, MCF-7, MDA-231 and MDA-468 breast cancer cell lines. At the same time, reduced (AA) and oxidized (DHA) forms of vitamin C uptake experiments were performed in all cell lines. We show here that human breast cancer tissues differentially express a form of SVCT2 transporter, that is systematically absent in normal breast tissues and it is increased in breast tumors. In fact, estrogen receptor negative breast cancer tissue, exhibit the most elevated SVCT2 expression levels. Despite this, our analysis in breast cancer cell lines showed that these cells are not able to uptake ascorbic acid and depend on glucose transporter for the acquisition of vitamin C by a bystander effect. This is consistent with our observations that this form of SVCT2 is completely absent from the plasma membrane and is overexpressed in mitochondria of breast cancer cells, where it mediates ascorbic acid transport. This work shows that breast cancer cells acquire vitamin C in its oxidized form and are capable of accumulated high concentrations of the reduced form. Augmented expression of an SVCT2 mitochondrial form appears to be a common hallmark across all human cancers and might have implications in cancer cells survival capacity against pro-oxidant environments.

Effects of long-term diet supplementation with Gliricidia sepium foliage mixed with Enterolobium cyclocarpum pods on enteric methane, apparent digestibility, and rumen microbial population in crossbred heifers1.

In the last decades, strategies have been evaluated to reduce rumen methane (CH4) production by supplementing tropical forages rich in secondary compounds; however, most of these beneficial effects need to be validated in terms of their persistence over time. The aim of this study was to assess CH4 emissions over time in heifers fed with and without Gliricidia sepium foliage (G) mixed with ground pods of Enterolobium cyclocarpum(E). Two groups of 4 crossbred (Bos taurus x Bos indicus) heifers (284 ±17 kg initial weight) were fed with 2 diets (0% and 15% of a mixture of the pods and foliage [E + G:0 and E + G:15, respectively]) over 80 d, plus 2 wk before the experiment, in which every animal was fed a legume and pod-free diet. Every 14 d, CH4 production, apparent digestibility, volatile fatty acids (VFA), and microbial population were quantified for each animal. The experiment was conducted with a repeated measurements design over time. Diets fed differed in terms of their crude protein (CP), condensed tannins, and saponins content supplied by E. cyclocarpum and G. sepium. For most of the experiment, dry matter intake (DMI) and digestible dry-matter intake (DDMI) were 6.3 kg DMI/d and 512 g DDMI/kg, respectively, for both diets (diet: P > 0.05). Apparent digestible crude protein (DCP) was reduced by 21 g DCP/kg DM when the diet was supplemented with E + G:15 (P = 0.040). Molar proportions of VFA's in the rumen did not differ between diets or in time (P > 0.05). Daily methane production, expressed in relation to DMI, was 23.95 vs. 23.32 g CH4/kg DMI for the diet E + G:0 and E + G:15, respectively (diet: P = 0.016; Time: P > 0.05). Percent gross energy loss as CH4 (Ym) with grass-only diets was above 8.1%, whereas when feeding heifers with the alternate supplementation, Ym values of 7.59% (P = 0.016) were observed. The relative abundance of total bacterial, protozoa, and methanogenic archaeal replicates was not affected by time nor by the incorporation of legume and pods into the diet (P > 0.05). Results suggest that addition of G. sepium mixed with E. cyclocarpum pods can reduce CH4 production in heifers and this response remains over time, without effect on microbial population and VFA concentration and a slight reduction in CPD digestibility.

Sustained blockade of ascorbic acid transport associated with marked SVCT1 loss in rat hepatocytes containing increased ascorbic acid levels after partial hepatectomy.

The liver has an extraordinary regenerative capacity in response to partial hepatectomy (PHx), which develops with neither tissue inflammation response nor alterations in the whole organism. This process is highly coordinated and it has been associated with changes in glutathione (GSH) metabolism. However, there are no reports indicating ascorbic acid (AA) levels after partial hepatectomy. AA and GSH act integrally as an antioxidant system that protects cells and tissues from oxidative damage and imbalance observed in a variety of diseases that affect the liver. Although rat hepatocytes are able to synthesize AA and GSH, which are the providers of AA for the whole organism, they also acquire AA from extracellular sources through the sodium-coupled ascorbic acid transporter-1 (SVCT1). Here, we show that hepatocytes from rat livers subjected to PHx increase their GSH and AA levels from 1 to 7 days post hepatectomy, whose peaks precede the peak in cell proliferation observed at 3 days post-hepatectomy. The increase in both antioxidants was associated with higher expression of the enzymes involved in their synthesis, such as the modifier subunit of enzyme glutamine cysteine ligase (GCLM), glutathione synthetase (GS), gulonolactonase (GLN) and gulonolactone oxidase (GULO). Importantly, rat hepatocytes, that normally exhibit kinetic evidence indicating only SVCT1-mediated transport of AA, lost more than 90% of their capacity to transport it at day 1 after PHx without evidence of recovery at day 7. This observation was in agreement with loss of SVCT1 protein expression, which was undetectable in hepatocytes as early as 2h after PHx, with partial recovery at day 7, when the regenerated liver weight returns to normal. We conclude that after PHx, rat hepatocytes enhance their antioxidant capacity by increasing GSH and AA levels prior to the proliferative peak. GSH and AA are increased by de novo synthesis, however paradoxically hepatocytes from rat subjected to PHx also suppress their capacity to acquire AA from extracellular sources through SVCT1.

Insulin Induces Relaxation and Decreases Hydrogen Peroxide-Induced Vasoconstriction in Human Placental Vascular Bed in a Mechanism Mediated by Calcium-Activated Potassium Channels and L-Arginine/Nitric Oxide Pathways.

HIGHLIGHTS Short-term incubation with insulin increases the L-arginine transport in HUVECs.Short-term incubation with insulin increases the NO synthesis in HUVECs.Insulin induces relaxation in human placental vascular bed.Insulin attenuates the constriction induced by hydrogen peroxide in human placenta.The relaxation induced by insulin is dependent on BKCa channels activity in human placenta. Insulin induces relaxation in umbilical veins, increasing the expression of human amino acid transporter 1 (hCAT-1) and nitric oxide synthesis (NO) in human umbilical vein endothelial cells (HUVECs). Short-term effects of insulin on vasculature have been reported in healthy subjects and cell cultures; however, its mechanisms remain unknown. The aim of this study was to characterize the effect of acute incubation with insulin on the regulation of vascular tone of placental vasculature. HUVECs and chorionic vein rings were isolated from normal pregnancies. The effect of insulin on NO synthesis, L-arginine transport, and hCAT-1 abundance was measured in HUVECs. Isometric tension induced by U46619 (thromboxane A2 analog) or hydrogen peroxide (H2O2) were measured in vessels previously incubated 30 min with insulin and/or the following pharmacological inhibitors: tetraethylammonium (KCa channels), iberiotoxin (BKCa channels), genistein (tyrosine kinases), and wortmannin (phosphatidylinositol 3-kinase). Insulin increases L-arginine transport and NO synthesis in HUVECs. In the placenta, this hormone caused relaxation of the chorionic vein, and reduced perfusion pressure in placental cotyledons. In vessels pre-incubated with insulin, the constriction evoked by H2O2 and U46619 was attenuated and the effect on H2O2-induced constriction was blocked with tetraethylammonium and iberiotoxin, but not with genistein, or wortmannin. Insulin rapidly dilates the placental vasculature through a mechanism involving activity of BKCa channels and L-arginine/NO pathway in endothelial cells. This phenomenon is related to quick increases of hCAT-1 abundance and higher capacity of endothelial cells to take up L-arginine and generate NO.

B7-H4(B7x)-Mediated Cross-talk between Glioma-Initiating Cells and Macrophages via the IL6/JAK/STAT3 Pathway Lead to Poor Prognosis in Glioma Patients.

PURPOSE: The objective of this study was to evaluate clinical significance and immunosuppressive mechanisms of B7-H4 (B7x/B7S1), a B7 family member, in glioma. EXPERIMENTAL DESIGN: B7-H4 levels in glioma tissue/cerebral spinal fluid (CSF) were compared between different grades of glioma patients. Survival data were analyzed with Kaplan-Meier to determine the prognostic value of B7-H4. Cytokines from CD133(+) cells to stimulate the expression of B7-H4 on human macrophages (Mφs) were investigated by FACS, neutralizing antibodies, and Transwell chemotaxis assay. shRNA, reporter vector, and chromatin immunoprecipitation were used to determine the binding of STAT3 to the B7-H4 promoter. The function of B7-H4(+) Mφs in vitro was evaluated through phagocytosis, T-cell proliferation/apoptosis, and cytokine production as well as in the xenografted model for in vivo analysis. RESULTS: We found that B7-H4 expression in tumors was associated with prognosis of human glioblastoma and correlated directly with malignant grades. Mechanistically, glioma initiating CD133(+) cells and Mφs/microglia cointeraction activated expression of B7-H4 via IL6 and IL10 in both tumor cells and microenvironment supporting cells. IL6-activated STAT3 bound to the promoter of B7-H4 gene and enhanced B7-H4 expression. Furthermore, CD133(+) cells mediated immunosuppression through B7-H4 expression on Mφs/microglia by silencing of B7-H4 expression on these cells, which led to increased microenvironment T-cell function and tumor regression in the xenograft glioma mouse model. CONCLUSIONS: We have identified B7-H4 activation on Mφs/microglia in the microenvironment of gliomas as an important immunosuppressive event blocking effective T-cell immune responses. Clin Cancer Res; 22(11); 2778-90. ©2016 AACR.

PP2A inhibition with LB100 enhances cisplatin cytotoxicity and overcomes cisplatin resistance in medulloblastoma cells.

The protein phosphatase 2A (PP2A) inhibitor, LB100, has been shown in pre-clinical studies to be an effective chemo- and radio-sensitizer for treatment of various cancers. We investigated effects associated with LB100 treatment alone and in combination with cisplatin for medulloblastoma (MB) in vitro and in vivo in an intracranial xenograft model. We demonstrated that LB100 had a potent effect on MB cells. By itself, LB100 inhibited proliferation and induced significant apoptosis in a range of pediatric MB cell lines. It also attenuated MB cell migration, a pre-requirement for invasion. When used in combination, LB100 enhanced cisplatin-mediated cytotoxic effects. Cell viability in the presence of 1 uM cisplatin alone was 61% (DAOY), 100% (D341), and 58% (D283), but decreased with the addition of 2 µM of LB100 to 26% (DAOY), 67% (D341), and 27% (D283), (p < 0.005). LB100 suppressed phosphorylation of the STAT3 protein and several STAT3 downstream targets. Also, LB100 directly increased cisplatin uptake and overcame cisplatin-resistance in vitro. Finally, LB100 exhibited potent in vivo anti-neoplastic activity in combination with cisplatin in an intracranial xenograft model.