Plant stem cell extract from Coffea canephora shows antioxidant, anti-inflammatory, and skin regenerative properties mediated by suppression of nuclear factor-κB.

Plant cell cultures have become a promising production platform of bioactive compounds for biomedical and cosmetic uses in the last decades. However, the success so far has been limited. The study aimed to evaluate the effectiveness of this unique biotechnology process to obtain a bioactive stem cell extract of Coffea canephora (SCECC) with antioxidant, anti-inflammatory, and regenerative properties. Total phenolic and flavonoid contents were determined in the SCECC by spectrophotometry. The chemical composition of the extracts was characterized by mass spectrometry. Antioxidant activity was evaluated using the colorimetric methods of free radical scavenging 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and the ferric reducing ability of plasma (FRAP). The anti-inflammatory activity was determined in lipopolysaccharide-stimulated RAW 264.7 macrophages through the production of superoxide anion (O2•-), nitric oxide (NO), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and the activity of nuclear factor kappa B (NF-κB). Moreover, the ability of SCECC to stimulate the proliferation and migration of fibroblasts was assessed. Five compounds were tentatively identified, two flavonoids, two phenolic acids, and one sugar. High phenolic content and antioxidant activity were observed in the SCECC. SCECC promoted the proliferation and migration of fibroblasts and suppressed the pro-inflammatory mediators O2•-, NO, TNF-α, and IL-6 in a dose-dependent manner. Moreover, SCECC inhibited the NF-κB transcription factor. Therefore, we obtained evidence that the extract from C. canephora stem cells can be used as a natural agent against skin damage. Hence, it could be of interest in cosmetics for preventing skin aging.

Plant stem cell extract from Coffea canephora shows antioxidant, anti-inflammatory, and skin regenerative properties mediated by suppression of nuclear factor-κB

Plant cell cultures have become a promising production platform of bioactive compounds for biomedical and cosmetic uses in the last decades. However, the success so far has been limited. The study aimed to evaluate the effectiveness of this unique biotechnology process to obtain a bioactive stem cell extract of Coffea canephora (SCECC) with antioxidant, anti-inflammatory, and regenerative properties. Total phenolic and flavonoid contents were determined in the SCECC by spectrophotometry. The chemical composition of the extracts was characterized by mass spectrometry. Antioxidant activity was evaluated using the colorimetric methods of free radical scavenging 2,2'-azinobis-3-ethylbenzothiazoline-6-sulfonic acid (ABTS) and the ferric reducing ability of plasma (FRAP). The anti-inflammatory activity was determined in lipopolysaccharide-stimulated RAW 264.7 macrophages through the production of superoxide anion (O2•-), nitric oxide (NO), tumor necrosis factor-alpha (TNF-α), interleukin-6 (IL-6), and the activity of nuclear factor kappa B (NF-κB). Moreover, the ability of SCECC to stimulate the proliferation and migration of fibroblasts was assessed. Five compounds were tentatively identified, two flavonoids, two phenolic acids, and one sugar. High phenolic content and antioxidant activity were observed in the SCECC. SCECC promoted the proliferation and migration of fibroblasts and suppressed the pro-inflammatory mediators O2•-, NO, TNF-α, and IL-6 in a dose-dependent manner. Moreover, SCECC inhibited the NF-κB transcription factor. Therefore, we obtained evidence that the extract from C. canephora stem cells can be used as a natural agent against skin damage. Hence, it could be of interest in cosmetics for preventing skin aging.

Fatty acid composition of vegetable oil blend and in vitro effects of pharmacotherapeutical skin care applications.

Vegetable oils have been used for a plethora of health benefits by their incorporation in foods, cosmetics, and pharmaceutical products, especially those intended for skin care. This study aimed to investigate the cutaneous benefits of a vegetable oil blend (VOB) formulation and its fatty acid composition. The anti-inflammatory activity was studied in macrophages of RAW 264.7 cells by investigating the release of nitric oxide (NO), superoxide anion generation (O2-), tumor necrosis factor-alpha (TNF-α), and interleukin 6 (IL-6). ABTS cation radical scavenging capacity assay, ferric reducing antioxidant potential (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and NO free radical scavenging assays were used to evaluate the antioxidant activity. VOB was tested for its ability to stimulate fibroblast proliferation and migration using the scratch assay, and antibacterial activity by the microdilution test. The fatty acid profile of a freshly prepared VOB formulation was determined by gas chromatography before and after accelerated stability testing. Chemical composition of VOB revealed the presence of oleic acid (C18:1n-9; 63.3%), linoleic acid (C18:2n-6; 4.7%), and linolenic acid (C18:3n-6; 5.1%) as major mono- and polyunsaturated fatty acids. No changes in the organoleptic characteristics and fatty acid composition were observed after the accelerated stability test. VOB 100 µg/mL reduced the healing time by increasing the total number of cells in the wounded area by 43.0±5.1% compared to the negative control group. VOB also suppressed the pro-inflammatory TNF-α and IL-6 cytokines, and NO and O2- production in lipopolysaccharide-stimulated macrophage cells. In conclusion, the VOB formulation contributed to the improvement of current therapeutic strategies for cutaneous applications in skin care.

Fatty acid composition of vegetable oil blend and in vitro effects of pharmacotherapeutical skin care applications

Vegetable oils have been used for a plethora of health benefits by their incorporation in foods, cosmetics, and pharmaceutical products, especially those intended for skin care. This study aimed to investigate the cutaneous benefits of a vegetable oil blend (VOB) formulation and its fatty acid composition. The anti-inflammatory activity was studied in macrophages of RAW 264.7 cells by investigating the release of nitric oxide (NO), superoxide anion generation (O2-), tumor necrosis factor-alpha (TNF-α), and interleukin 6 (IL-6). ABTS cation radical scavenging capacity assay, ferric reducing antioxidant potential (FRAP), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and NO free radical scavenging assays were used to evaluate the antioxidant activity. VOB was tested for its ability to stimulate fibroblast proliferation and migration using the scratch assay, and antibacterial activity by the microdilution test. The fatty acid profile of a freshly prepared VOB formulation was determined by gas chromatography before and after accelerated stability testing. Chemical composition of VOB revealed the presence of oleic acid (C18:1n-9; 63.3%), linoleic acid (C18:2n-6; 4.7%), and linolenic acid (C18:3n-6; 5.1%) as major mono- and polyunsaturated fatty acids. No changes in the organoleptic characteristics and fatty acid composition were observed after the accelerated stability test. VOB 100 µg/mL reduced the healing time by increasing the total number of cells in the wounded area by 43.0±5.1% compared to the negative control group. VOB also suppressed the pro-inflammatory TNF-α and IL-6 cytokines, and NO and O2- production in lipopolysaccharide-stimulated macrophage cells. In conclusion, the VOB formulation contributed to the improvement of current therapeutic strategies for cutaneous applications in skin care.

[Influence of zinc administered by total parenteral nutrition on plasmatic zinc levels, on reactive C protein, on serum interleukin-6 and on serum interleukin-6 soluble receptor, in critical patients]. / Influencia del zinc administrado a pacientes críticos con nutrición parenteral sobre los niveles de zinc plasmático, proteína C reactiva, interleuquina-6 y receptor soluble de interleuquina-6.

OBJECTIVES: To study the interrelationship between serum Interleukin-6 (IL-6), serum Interleukin-6 soluble Receptor (IL-6 sR), C-Reactive Protein (C-RP), plasmatic Zinc levels (PlZn) and their response in relation to Zn administered by TPN, in critical patients. METHODS: 17 patients, receiving TPN as a consequence of acute pancreatitis (n = 4) or after a major abdominal surgery due to intestinal cancer (n = 7), intestinal fístula (n = 3), intestinal obstruction (n = 2) or intestinal íleus (n = 1) were studied. At the beginning (To) and at the end of the TPN administration (6-21 days) serum IL-6 and IL-6 sR were determined by ELISA; C-RP ultrasensitive (C-RP us) by inmunoturbidimetric method; Zn was determined in TPN and in plasma by Atomic Absorption Spectrometry. Characteristics of the patients were (mean +/- SD and ranges): age: 60.6 +/- 11.7 (37-77) years; BMI (kg/m(2)): 26.0 +/- 3.4 (19.9-34.0). RESULTS: The results (mean +/- standard deviation and ranges) were: Zn provided by TPN (mg/d): 6.1 +/- 2.0 (range 2.8 to 10.8). Biochemical levels were, at To and Tf, respectively: (mean+/-SD and ranges) were at To y Tf, respectively: Zn Pl (microg/dl): 104 +/- 46 (35-177); 120 +/- 55 (52-229); IL-6 (pg/mL) 93 +/- 74 (10-262); 117 +/- 180 (7-761); IL6sR (pg/mL): 1,012 +/- 322 (589-1855); 1,269 +/- 451 (631-2195); C-RP us (mg/L): 71 +/- 63 (2-196); 65 +/- 43 (0-137). There was no correlation between variations of IL6, IL6sR, C-RP, PlZn levels and the daily amount of Zn administered in the TPN mixtures. Two patients presented a bad evolution; they received 4.2 and 5.2 md/d of Zn and showed an increase of IL6 levels, maintained high levels of IL6sR but C-RP levels decreased. CONCLUSIONS: the range of 2.8 to 10.8 mg/d of Zn administered in TPN mixtures did not exacerbate the inflammatory response.

[Relationship between the amount of copper and zinc given to critically ill patients on total parenteral nutrition and plasma and erythrocyte copper and zinc levels]. / Relación entre las cantidades de cobre y zinc administradas a pacientes graves con nutrición parenteral total y los niveles de cobre y zinc en plasma y eritrocitos.

OBJECTIVES: Complications resulting from zinc and copper deficiency, or adverse effects from excessive zinc and copper intake should be avoided during total parenteral nutrition (TPN). This study was conducted on critically ill patients requiring TPN to determine the relationship between the zinc and copper levels of the TPN mixtures, patients' clinical progression, and changes in plasma levels of zinc, serum levels of copper, and erythrocyte levels of zinc and copper. PATIENTS AND METHODS: 29 adult critically ill patients following pancreatitis or after a major abdominal surgery were studied. Zinc and copper levels in TPN, plasmatic zinc levels, copper serum levels and erythrocyte levels of zinc and copper were determined at the onset and at the end of the treatment (5-21 days) (using Atomic Absorption Spectrometry). RESULTS: The mean+/-standard deviation (and ranges in parenthesis) of zinc and copper levels in TPN were (microg/mL): zinc: 4.2+/-1.7 (1.8 a 9.3); copper: 0.94+/-0.66 (0.1 a 3.1). Biochemical parameters at the onset and at the end of the treatment were, respectively: (microg/mL): plasmatic zinc: 80+/-45 (29-205) and 122+/-56 (37-229); erythrocyte zinc: 2,300+/-1,070 (790-5,280) and 2,160+/-920 (790-4,440); serum copper (microg/dL): 124+/-35 (62-211) and 128+/-41 (60- 238); erythrocyte copper (microg/dl): 72+/-39 (4-183) and 70+/-41 (9-156). Plasmatic and erythrocyte zinc levels did not correlated neither at the onset nor at the end of the treatment. Changes in erythrocyte zinc levels correlated with daily administered zinc (mg/d) in the parenteral nutrition (r=0.38). Serum copper and erythrocyte copper showed significant correlation at the onset (p=0.0005) and at the end of treatment (p=0.008). Changes of serum or erythrocyte copper levels showed a significant correlation with daily administered copper (r=0.31 and 0.26, respectively). CONCLUSIONS: These results show that: 1) determination of erythrocyte zinc and copper levels in these critically ill patients would help to control zinc and copper administration levels in TPN and avoid adverse effects; 2) TPN Zn levels should be less than 8.1 mg/d, and 3) TPN Cu levels higher than 1.2 mg/d would not be recommended.

Active circulation of bluetongue vaccine virus serotype-2 among unvaccinated cattle in central Italy.

Several seroconversions occurring in 2002 among sentinel cattle during the bluetongue-vaccination campaign in Lazio and Tuscany (central Italy) led to the suspicion of vaccine-virus circulation. Therefore in 2003, 17 seroconverting sentinel herds were investigated for the characteristics of the virus involved. From these farms, 91 unvaccinated animals and 57 Culicoides pools were tested for the presence of the bluetongue vaccine virus (serotype-2) or other strains. The presence of vaccine virus serotype-2 was confirmed by PCR followed by restriction analysis in the whole blood of 17 unvaccinated sentinel cattle and 12 pools of Culicoides imicola or C. obsoletus. Of the 17 herds, five were positive only for vaccine virus serotype-2, four were positive for other strains and two for both the vaccine and other strains; the remaining premises were virologicaly negative. The vaccine virus serotype-2 also was detected in areas not included in the vaccination campaign.

Association between the 2001-2003 bluetongue epidemic in Lazio and Tuscany (central Italy) and distribution and abundance of Culicoides imicola and C. obsoletus vectors.

During the epidemic of bluetongue (BT) in Lazio and Tuscany between 2001 and 2003, the distribution pattern of Culicoides imicola did not always correspond either geographically or seasonally, with virus circulation. Culicoides obsoletus was observed to be abundant, ubiquitous and active throughout the year. The geographical and seasonal distribution of BT virus (BTV), C. imicola and C. obsoletus was compared. The territory of the two regions was divided into 30 cells each measuring 1 600 km(2). The presence of C. obsoletus was recorded in every cell, while C. imicola was detected in 18 of the 30 cells, but was absent in 6 of the 21 cells that indicated the presence of BTV. The occurrence of seroconversions appeared to be positively correlated with maximum C. obsoletus catches. Seroconversions were recorded throughout the year, even when C. imicola was not active, whereas C. obsoletus was detected during the entire period. The occurrence of BTV circulation in areas and periods where C. imicola was absent, and the abundant and constant presence of adult C. obsoletus in all the cells, suggest the active role of the latter species in BTV circulation in central Italy.