MMP-2 and MMP-9 secretion is elevated in several types of human cancers and their elevated expression has been associated with poor prognosis. Expression of MMPs is highly regulated by cytokines and ...signal transducation pathways, including those activated by phorbol 12-myristate 13-acetate (PMA). The aim of this study was to examine the effect of PMA on MMP-2 and MMP-9 secretion in 42 different human cancer cell lines, selected on the basis of their organ malignancies. They were cultured in the recommended media supplemented with 10% FBS and antibiotics in 24-well tissue culture plates. At near confluence, the cells were washed with PBS, 0.5 ml of medium was added, and the cultures were incubated. Parallel sets of cultures were also treated with PMA for induction of enzymes. After 24 h the media were collected and MMP-2 and MMP-9 levels were assayed by gelatinase zymography. Based on MMP-2 and MMP-9 secretion without and with PMA treatment, the various human cancer cell lines fell into one of two major groups. The first group characterized by low basal MMP-9 secretion fell into three different categories of susceptibility to PMA induction of MMP-9 expression: resistant, moderately susceptible and highly susceptible. High basal MMP-9 levels responsive to PMA induction characterized the second group. Most cancer cell lines examined exhibited basal levels of MMP-2, MMP-9 or both. MMP-2 secretion was not induced by PMA in any of the cancer cells examined.
Matrix metalloproteinases (MMPs) secreted by cervical and ovarian cancer, especially MMP-2 and MMP-9, play crucial roles in tumor invasion and metastasis. We examined the effect of cytokines, ...mitogens, inducers and inhibitors on MMP-2 and MMP-9 expression in cervical and ovarian cancer cell lines. Human cervical (HeLa and DoTc2-4510) and ovarian (SK-OV-3) cell lines were cultured in appropriate media. At near confluence, the cells were washed with PBS and incubated in serum-free medium with various concentrations of several cytokines, mitogens and inhibitors. After 24 h the media were removed and analyzed for MMP-2 and MMP-9 by gelatinase zymography and quantitated by densitometry. HeLa and SK-OV-3 cell lines expressed MMP-2 whereas DoTc2-4510 cells expressed MMP-9. Treatment of cervical cancer cell lines (HeLa and DoTc2-4510) with PMA had no effect on MMP-2 expression and a moderate stimulatory effect in ovarian cancer cell line SK-OV-3. MMP-9 was stimulated by phorbol 12-myristate 13-acetate in HeLa cells and enhanced in DoTc2-4510. Tumor necrosis factor-alpha and interleukin-1beta, had slight inhibitory effect on HeLa cell expression of MMP-2 while lipopolysaccharide stimulated MMP-2 in HeLa cells. Doxycycline, epigallocatechin gallate, a nutrient mixture, actinomycin-D, cyclohexamide, retinoic acid and dexamethasone inhibited MMP-2 in HeLa and SK-OV-3 cell lines and inhibited MMP-9 in DoTc2-4510. Our results show that cytokines, mitogens, inducers and inhibitors have an up or down regulatory effect on MMP-2 and MMP-9 expression in ovarian and cervical cancer cell lines, suggesting these agents may be effective strategies to treat these cancers.
Type IV collagenase matrix metalloproteinases (MMPs), especially MMP-2 and MMP-9, have been found to promote invasion and metastasis of malignant tumors. Extracellular matrix (ECM) degradation by ...MMPs and increased expression of MMPs in cancer cells and tumor microvascular endothelial cells make MMPs an attractive target for cancer. Focused on a common pathomechanism of cancer growth and invasion, the disintegration of connective tissue, we used natural approaches to increase the integrity and strength of connective tissues. Utilizing the principle of nutrition synergy, we developed a novel micronutrient mixture (NM) containing lysine, proline, ascorbic acid and green tea extract. This study evaluates the potency of the components EGCG and green tea extract independently compared to that of NM on modulation of patterns of MMP-2 and MMP-9 expression in four cancer cell lines expressing MMP-2, MMP-9 or both. Human fibrosarcoma (HT-1080), hepatocellular carcinoma (SK-Hep-1), glioblastoma (T-98G), uterine leiomyosarcoma (SK-UT-1) cell lines were obtained from ATCC and grown in minimum essential medium (MEM) supplemented with 10% FBS, penicillin (100 U/ml) and streptomycin (100 mg/ml) in 24-well tissue culture plates. At near confluence, the cells were treated with agents dissolved in media and tested at concentrations indicated in triplicate at each dose. Cells were also treated with PMA 100 ng/ml to study enhanced expression of MMP-9. MMP expression was assessed by gelatinase zymography. Fibrosarcoma and hepatocellular carcinoma cells expressed both MMP-2 and MMP-9. Glioblastoma cells expressed MMP-2 and PMA treatment induced MMP-9 expression. Uterine leimyosarcoma cells expressed no MMPs but PMA induced MMP-9. NM was the most potent dose-dependent inhibitor of MMPs, followed by green tea extract and EGCG. In conclusion, these results suggest the enhanced efficacy of nutrients working in synergy to modulate complex pathways such as MMP expression.
This study aimed at evaluating the effect of increasing organic loading rates and of enzyme pretreatment on the stability and efficiency of a hybrid upflow anaerobic sludge blanket reactor (UASBh) ...treating dairy effluent. The UASBh was submitted to the following average organic loading rates (OLR) 0.98
Kg.m
−3.d
−1, 4.58
Kg.m
−3.d
−1, 8.89
Kg.m
−3.d
−1 and 15.73
Kg.m
−3.d
−1, and with the higher value, the reactor was fed with effluent with and without an enzymatic pretreatment to hydrolyze fats. The hydraulic detention time was 24
h, and the temperature was 30
±
2
°C. The reactor was equipped with a superior foam bed and showed good efficiency and stability until an OLR of 8.89
Kg.m
−3.d
−1. The foam bed was efficient for solid retention and residual volatile acid concentration consumption. The enzymatic pretreatment did not contribute to the process stability, propitiating loss in both biomass and system efficiency. Specific methanogenic activity tests indicated the presence of inhibition after the sludge had been submitted to the pretreated effluent. It was concluded that continuous exposure to the hydrolysis products or to the enzyme caused a dramatic drop in the efficiency and stability of the process, and the single exposure of the biomass to this condition did not inhibit methane formation.
► Anaerobic treatment of lids rich effluents is feasible but requires optimization. ► Lipids can cause sludge floatation in granular reactors and clogging in fixed bed. ► The enzymatic pre-treatment was studied in a UASBh treating dairy wastewater. ► Solids retention and volatile acid consumption occurred in the superior foam bed. ► Enzymatic pretreatment caused drop in the efficiency and stability of the process.
Invasion of surrounding tissues by malignant cells is a complex process mediated by the matrix degrading enzymes. In many solid tumors, the expression of MMPs, especially MMP-2 and MMP-9, is higher ...in stromal cells than in the tumor cells, suggesting stromal cells as the major source of these enzymes. Cytokines and signal transduction pathways, including those activated by phorbol 12-myristate 13-acetate (PMA), regulate the expression of MMPs. The aim of this study was to examine the pattern of MMP-2 and MMP-9 expression in human normal cells and in PMA-treated cells to determine if specific patterns of expression were associated with tissues of different origin. Epithelial, connective, and muscle tissues were selected since carcinomas, sarcomas, and adenosarcomas are derived from these tissue types, respectively. The cell lines were cultured in their recommended media and supplemented with 10% FBS and antibiotics in 24-well tissue culture plates. At near confluence, the cells were washed and fresh medium added. A parallel set of cultures was treated with PMA. After 24 h of incubation, media were collected and analyzed for MMP-2 and MMP-9 by gelatinase zymography. The results indicate that the normal cell expression of MMP-2 and MMP-9 depends on their primary tissue subtype. All cell lines, regardless of tissue origin, expressed MMP-2. PMA induced MMP-9 expression in glandular epithelia, supportive connective tissue, and muscle tissue cell lines. However, cell lines of endothelial origin and proper connective tissue were insensitive to PMA. These results suggest that MMP-2 and MMP-9 are differentially regulated and an understanding of this may open up avenues to use these enzymes as targets for therapy.
Matrix metalloproteinase (MMP)-2 and -9 secretion is elevated in various human cancers and their elevated expression has been associated with poor prognosis due to associated increased cancer cell ...invasion and metastasis.
To examine the correlation between in vitro MMP-2 and MMP-9 secretion and Matrigel invasion in 42 different human cancer cell lines (selected on the basis of organ malignancies) treated with a nutrient mixture (NM).
The cells were cultured in their recommended media supplemented with 10% FBS and antibiotics in 24-well tissue culture plates. At near confluence, the cells were treated with NM dissolved in media at 0, 10, 50, 100, 500 and 1000 μg/mL in triplicate. Parallel sets of cultures were also treated with phorbol 12-myristate 13-acetate (PMA) 100 ng/mL for induction of enzymes. After 24 h the media were collected and MMP-2 and MMP-9 levels were assayed by gelatinase zymography. Invasion studies were conducted using Matrigel in 24-well plates.
Correlation of pooled data from different cancer cell line groups demonstrated dose-dependent inhibition of MMP-2 and -9 and Matrigel invasion with NM treatment and significant negative correlation between MMP-2 and MMP-9 levels and Matrigel invasion. Pooled data of cell lines expressing only MMP-2 and resistance to PMA induction of MMP-9 showed significant negative correlation (r = -0.77, p = 0.003) between MMP-2 secretion and inhibition of invasion through Matrigel. Cell lines expressing only MMP-9, showed significant negative correlation (r = -0.726, p = 0.003) between MMP-9 secretion and Matrigel invasion. Pooled data of cell lines expressing MMP-2 and MMP-9 demonstrated significant negative correlation (r = -0.821, p < 0.0001) between MMP-9 secretion and inhibition of invasion through Matrigel. Pooled data of cancer cell lines expressing no basal MMP- 9 secretion demonstrated significant negative correlation (r = -0.686, p < 0.0001) between PMA-induced MMP-9 secretion and inhibition of invasion through Matrigel.
In conclusion, regardless of MMP-2 and MMP-9 patterns of expression, MMP modulation by NM was found to be significantly correlated with NM modulation of Matrigel invasion of these cell lines.
Policy Points
Models for access to care for uninsured immigrant children that mitigate structural and sociopolitical barriers to inclusive health care include funding structures (e.g., ...state‐sponsored coverage) and care delivery systems (e.g., federally qualified health centers,).
Although the quintessential model of access to care necessitates health coverage for all children regardless of immigration status or date of United States entry, incremental policy change may more realistically and efficiently advance equitable access to high‐quality health care.
Intentional advocacy efforts should prioritize achievable goals that are grounded in data, are attentive to the sociopolitical milieu, are inclusive of diverse perspectives, and would meaningfully impact health care access and outcomes.
The Parent Attitudes about Childhood Vaccines (PACV) survey is a validated instrument for identifying vaccine-hesitant parents; however, a Spanish version is not available. Utilizing the WHO ...framework for translating survey instruments, we used an iterative process for developing the Spanish PACV that included forward translation, expert panel review, back translation and pre-testing that utilized cognitive interviewing. We made revisions to the Spanish PACV at each step, focusing on addressing inclusivity, readability, clarity and conceptual equivalence. The expert panel was comprised of 6 Spanish-speaking medical and research professionals who worked alongside 3 study team members. Pre-testing was conducted using convenience sampling of Spanish-speaking parents (N = 35) who had a child receiving care at the residents' continuity clinic at Texas Children's Hospital. Most pre-testing participants were married (80.6%), mothers (97.1%), ≥30 years of age (88.2%) and had a high school education or less (70.6%). While the majority of participants stated the survey was easy to complete, the translation of 5 PACV items was further revised to improve interpretability. We conclude that the final Spanish PACV is conceptually equivalent and culturally appropriate for most Hispanic populations.
Background Multiple Sclerosis (MS) is a common neurological disease among white populations of European origin. Frequencies among Latin Americans continue to be studied, however, epidemiologic, and ...clinical characterization studies lack from Central American and Caribbean countries. Ethnicity in these countries is uniformly similar with a prevalent Mestizo population. Methods and results Data from January 2014 to December 2019 from Guatemala, El Salvador, Honduras, Nicaragua, Costa Rica, Panama, Dominican Republic, and Aruba on demographic, clinical, MRI and phenotypic traits were determined in coordinated studies: ENHANCE, a population-based, retrospective, observational study on incidence and clinical characteristics, and from the subgroup with MS national registries (Aruba, Dominican Republic, Honduras, and Panama), data on prevalence, phenotypes and demographics. Expanded Disability Status Scale (EDSS), and therapeutic schemes were included. ENHANCE data from 758 patients disclosed 79.8% of Mestizo ethnicity; 72.4% female; median age at onset 31.0 years and 33.2 at diagnosis. The highest incidence rate was from Aruba, 2.3–3.5 × 100,000 inhabitants, and the lowest, 0.07–0.15 × 100,000, from Honduras. Crude prevalence rates per 100,000 inhabitants fluctuated from 27.3 (Aruba) to 1.0 (Honduras). Relapsing MS accounted for 87.4% of cases; EDSS <3.0 determined in 66.6% (mean disease duration: 9.1 years, SD ± 5.0); CSF oligoclonal bands 85.7%, and 87% of subjects hydroxyvitamin D deficient. Common initial therapies were interferon and fingolimod. Switching from interferon to fingolimod was the most common escalation step. The COVID-19 pandemic affected follow-up aspects of these studies. Conclusion This is the first study providing data on frequencies and clinical characteristics from 8 countries from the Central American and Caribbean region, addressing MS as an emergent epidemiologic disorder. More studies from these areas are encouraged.
Vitamin A status in Cuban children aged 6–11 years Macías-Matos, Consuelo; Pita-Rodríguez, Gisela; Monterrey-Gutiérrez, Pedro ...
Public health nutrition,
01/2008, Volume:
11, Issue:
1
Journal Article
Peer reviewed
Open access
A nationwide study was performed in Cuba to assess vitamin A status and the intake of vitamin-A-providing foods in children aged 6-11 years.
The sample comprised 1191 schoolchildren from first to ...sixth grade, both sexes, from municipalities randomly selected from the five eastern provinces of Cuba in 2002 (first semester) and from the four western and four central provinces in 2003 (first semester). A food-frequency questionnaire was completed by 2038 mother-and-child pairs.
Mean (+/-standard deviation) plasma retinol concentrations were 1.77 +/- 0.48 micromol l-1 in the western, 2.01 +/- 0.56 micromol l-1 in the central and 1.40 +/- 0.41 micromol l-1 in the eastern region. No child had plasma retinol concentration below 0.35 micromol l-1, indicative of a high risk of clinical deficiency. Subclinical deficiency, plasma retinol concentration of 0.35-0.7 micromol l-1, was seen in 1.05 micromol l-1) was present in >90% of subjects in all western and central provinces, and in one of the eastern provinces (Holguín), whereas in the four remaining eastern provinces, adequate status was present in >75%. Only nine fruits and vegetables were consumed frequently (>3 times per week) by >50% of children. Thirty-seven per cent regularly consumed a supplement containing vitamin A.
Most Cuban children aged 6-11 years had adequate vitamin A status. Consumption of foods rich in vitamin and provitamin A, especially vegetables, was frequent but limited to a small variety of foods.
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