In Vitro Effects of Calcium Fructoborate on fMLP-stimulated Human Neutrophil Granulocytes

Discovery of naturally occurring boron complexes with organic compounds containing hydroxyl groups, sugars, and polysaccharides, adenosine-5-phosphate, pyridoxine, riboflavin, dehydroascorbic acid, and pyridine nucleotides led to the reassessment of the biochemical role of boron. Boron’s anti-inflammatory actions were claimed but not yet demonstrated. This study investigated the effects of calcium fructoborate (CF) on the human polymorphonuclear neutrophils (PMN) that play a central role in the inflammatory response. Our results demonstrated that CF exposure induced a dose-dependent decrease in cell viability. Treatment of PMN cells, for 24 h, with 22,500 μM CF led to a decrease in cell viability by 61.1%, an inhibition of respiratory burst by 92.9% in the case of fMLP-stimulated cells, a diminution of intracellular level of superoxide anion with 59.3%, and a stimulation of superoxide dismutase activity by 72% in unstimulated PMN cells. Altogether, these results suggest the antioxidant and anti-inflammatory properties of CF.     

Comparative Effects of Boric Acid and Calcium Fructoborate on Breast Cancer Cells

Recent studies suggested that boron has a chemo-preventive role in prostate cancer. In the present report, we investigated the effects of calcium fructoborate (CF) and boric acid (BA) on activation of the apoptotic pathway in MDA-MB-231 human breast cancer cells. Exposure to BA and CF inhibited the proliferation of breast cancer cells in a dose-dependent manner. Treatment with CF but not BA resulted in a decrease in p53 and bcl-2 protein levels. Furthermore, after the treatment with CF, augmentation of pro-caspase-3 protein expression, cytosolic cytochrome c level, and caspase-3 activity were observed, indicating apoptotic cell death induction. This was also demonstrated by terminal deoxynucleotidyl transferase-mediated 2′-deoxyuridine 5′-triphosphate nick-end-labeling assay. In conclusion, our data provide arguments to the fact that both BA and CF inhibited the growth of breast cancer cells, while only CF induced apoptosis. Additional studies will be needed to identify the underlying mechanism responsible for the observed cellular responses to these compounds and to determine if BA and CF may be further evaluated as chemotherapeutic agents for human cancer.     

Experimental and Theoretical Studies of Calcium Fructoborate

Calcium fructoborate samples of composition Ca(C₆H₁₀O₆BO)₂·3.5H₂O were characterized by chemical analysis, infrared and Raman spectroscopy, and thermoanalytical (thermogravimetric and differential thermal analysis) data. Theoretical studies, using density functional theory, were made for seven different structural models of the fructoborate moiety, and the most stable structure could be derived from these calculations. The results of the theoretical study also allow improving the assignment of the vibrational spectra of the compound.     

In Vitro Effects of Calcium Fructoborate upon Production of Inflammatory Mediators by LPS-stimulated RAW 264.7 Macrophages

The present study is supported by our previous findings suggesting that calcium fructoborate (CF) has anti-inflammatory and antioxidant properties. Thus, we investigated the effects of CF on a model for studying inflammatory disorders in vitro represented by lipopolysaccharide (LPS)-stimulated murine macrophage RAW 264.7 cells. This investigation was performed by analyzing the levels of some mediators released during the inflammatory process: cytokines such as tumor necrosis factor-α (TNF-α), interleukins IL-1β and IL-6 as well as cyclooxygenase-2 (COX-2), the main enzyme responsible for endotoxin/LPS-induced prostaglandin synthesis by macrophages. We also measured production of nitric oxide (NO) that plays an important role in the cytotoxicity activity of macrophages towards microbial pathogens. After CF treatment of LPS-stimulated macrophages we found an up-regulation of TNF-α protein level in culture medium, no significant changes in the level of COX-2 protein expression and a decrease in NO production as well as in IL-1β and IL-6 release. Collectively, this series of experiments indicate that CF affect macrophage production of inflammatory mediators. However, further research is required in order to establish whether CF treatment can be beneficial in suppression of pro-inflammatory cytokine production and against progression of endotoxin-related diseases.     

Calcium Fructoborate Helps Control Inflammation Associated with Diminished Bone Health

Inflammation has been identified as a possible contributory factor to disruption of the normal bone remodeling process, a process essential to healthy bone mineral density. Several large population-based clinical studies have specifically shown that levels of C-reactive protein, an immune recognition protein that is a sensitive marker of inflammation, are inversely and independently associated with total bone mineral density. The evidence suggests that control of C-reactive protein levels may contribute to bone health by protecting against inflammation’s disruption of the equilibrium between bone resorption and bone deposition. Calcium fructoborate, a patented complex of calcium, fructose, and boron found naturally in fresh and dried fruits, vegetables and herbs, and wine, is a sugar-borate ester. A growing body of peer-reviewed, published clinical research indicates that the calcium fructoborate significantly reduces serum levels of the C-reactive protein in humans, suggesting that this unique plant–mineral complex may contribute to bone health by controlling the inflammation associated with loss of bone mineral density.     

A Double-Blind, Placebo-Controlled Pilot Study to Evaluate the Effect of Calcium Fructoborate on Systemic Inflammation and Dyslipidemia Markers for Middle-Aged People with Primary Osteoarthritis

The objective of this pilot study was to determine whether 15?days of dietary supplementation with calcium fructoborate could acutely modulate inflammatory and lipid blood markers in individuals diagnosed with primary osteoarthritis. During 2?weeks, a placebo-controlled, randomized, double-blind study was conducted on 116 subjects that were initially recruited. Seventy-two subjects started the study, being divided into four groups, and only 60 completed the study as designed. The aim was to compare the effects of calcium fructoborate to placebo on subjects diagnosed with knee primary osteoarthritis. The obtained outcomes were inflammation biomarkers (C-reactive protein, fibrinogen, and erythrocyte sedimentation rate) and lipid markers (triglycerides, total cholesterol, LDL cholesterol, and HDL cholesterol). No serious adverse events were reported. The calcium fructoborate showed beneficial effect on the inflammatory markers for all groups subjected to the treatment when compared with the placebo group and slight changes in the lipid metabolism. This study suggests that short-term (2?weeks) calcium fructoborate supplementation in patients with osteoarthritis symptoms has a favorable prognosis on inflammation diseases.     

天然大豆硒蛋白抗肿瘤作用研究

建立S180小鼠肿瘤动物模型,设计用不同的剂量处理,采用胃饲法补充从高硒土栽培大豆中提取的硒蛋白,研究大 豆硒蛋白的抗肿瘤活性。结果表明:在202.50μg(Se)/kg(体重)剂量范围内,大豆硒蛋白剂量增加可显著加强对S180肉瘤生 长的抑制,并能推迟S180肉瘤大鼠的死亡时间,补硒剂量在202.50μg/kg时对肿瘤生长抑制率达78%左右;相关指标显示大豆 硒蛋白可能是通过提高机体的抗氧化能力、调节免疫功能而发挥抗癌作用。     

Bowman-Birk蛋白酶抑制剂的体外和动物模型实验抗癌实验研究进展

Bowman-Birk蛋白酶抑制剂（BBI）是一种植物丝氨酸蛋白酶抑制剂,能同时抑制胰蛋白酶和胰凝乳蛋白酶。体外及动物实验都证实：BBI纯品或BBI浓缩物均有较强的抗癌活性。本文综述了近年来有关BBI的一些抗癌研究进展,包括BBI的结构、临床前研究及可能的分子机制等。     

An Experimental Study of Calcium Acquisition and its Effects on the Calcifuge Moss Pleurozium schreberi

In a field experiment to investigate the sources and effectson growth of Ca in the calcifuge moss Pleurozium schreberi,significant quantities of Ca reached the growing shoot apicesfrom a CaCO₃ layer placed on the mineral soil surface Top applicationsof 0.5 and 5 mol m^–3 CaCl₂ raised the exchangeable andintracellular Ca concentrations and displaced natural exchangeableK and Mg The 5 mol m^–3 CaCl₂ treatment also caused a significantreduction in intracellular Mg indicating that Mg uptake is dependenton an initial exchange step No growth differences were notedbetween treatments, possibly because ionic changes had not reacheda detrimental level within the 28 weeks of the experiment ina second experiment, shoot apices of Pleurozium schreberi, Pseudoscleropodiumpurum and Calliergon cuspidatum were grown on nylon gauze underintermittent distilled-water mist At weekly intervals the shootswere saturated with CaCl₂ solutions providing factorial combinationsof Ca and pH Growth of C cuspidatum and P purum from chalk soilwas reduced at high (0.01) Ca concentration whereas Pleuroziumschreberi and Pseudoscleropodium purum from acidic clay wereunaffected The pH treatments did not significantly affect mossgrowth Initial tissue levels of K and Mg were lower in the mossesfrom chalk and it is suggested that the CaCl₂ treatments causednutrient deficiencies in these plants Mosses from acidic soilcontained less exchangeable Ca than the chalk plants and grewpoorly in the absence of CaCl₂, perhaps due to the developmentof Ca deficiency Bryophyte growth, calcium uptake, pH, mineral nutrition, Pleurozium schreberi, Pseudoscleropodium purum, Calliergon cuspidatum     

Anticarcinogenic Alkaloids in the Cultured Cells of Cephalotaxus fortunei

Calli were induced from the leaves and stems of Cephalotaxus fortunei Hook. f. on MS medium supplemented with 0. 1 mg/L KT and 3 mg/L NAA, and from which the suspension culture cell line of this plant was established for the first time. Factors such as light, pH value of the medium, concentration of plant hormone, carbon resources and addition of substances to the medium, which affect the growth of suspension cells were investigated. The results showed that suspension cells grew appropriately at pH 5.8 with a low concentration of sucrose or glucose, and a low level of NAA. No difference effect on cell growth was seen between sucrose and glucose. Phenylalanine and protein hydrolysate were not suitable for cell growth in suspension cultures, and light inhibited cell growth. A sensitive and rapid high-performance liquid chromatographic method has been developed for detecting the alkaloids in cultured cells. The results revealed the following contents of cephalotaxine and its anticancer esters in cultured cells: harringtonine, isoharringtonine and homoharringtonine. The total alkaloid production in cell suspension cultures was doubled as that in solid cultures. The relative amounts of cephalotaxine, drupacine, harringtonine, homoharringtonine and isoharringtonine in suspension cells was 22%, 6%, 8%, 23% and 41% respectively. In addition, other alkaloid as deoxyharringtonine and some steroids, including ergdst-5-en-3-ol. stigmasta-5, 22-dien-3-ol, β-sitosterin and 2-naphthalenamine have also been detected in cell cultures using GC/MS combined technique.     

Effects of an Anticarcinogenic Bowman-Birk Protease Inhibitor on Purified 20S Proteasome and MCF-7 Breast Cancer Cells

Proteasome inhibitors have been described as an important target for cancer therapy due to their potential to regulate the ubiquitin-proteasome system in the degradation pathway of cellular proteins. Here, we reported the effects of a Bowman-Birk-type protease inhibitor, the Black-eyed pea Trypsin/Chymotrypsin Inhibitor (BTCI), on proteasome 20S in MCF-7 breast cancer cells and on catalytic activity of the purified 20S proteasome from horse erythrocytes, as well as the structural analysis of the BTCI-20S proteasome complex. In vitro experiments and confocal microscopy showed that BTCI readily crosses the membrane of the breast cancer cells and co-localizes with the proteasome in cytoplasm and mainly in nucleus. Indeed, as indicated by dynamic light scattering, BTCI and 20S proteasome form a stable complex at temperatures up to 55°C and at neutral and alkaline pHs. In complexed form, BTCI strongly inhibits the proteolytic chymotrypsin-, trypsin- and caspase-like activities of 20S proteasome, indicated by inhibition constants of 10⁻⁷ M magnitude order. Besides other mechanisms, this feature can be associated with previously reported cytostatic and cytotoxic effects of BTCI in MCF-7 breast cancer cells by means of apoptosis.     

Hormonal Effects on Calcium Metabolism in Crustacea

Cyclic shifts of calcium in the exoskeleton and soft tissues,as they are related to the intermolt cycle in crayfish, arereviewed. Regulatory factors, derived from the eyestalk, influencelevels of exoskeletal calcium; eyestalk extracts prepared fromanimals in premolt decrease shell calcium, while reciprocallyextracts from animals in intermolt increase it when these hormonalsources are injected into animals in the premolt stage (D₀-D₄). In addition, premolt eyestalk extract results in an increasein gastrolith calcium. In the exchange of calcium between theanimal and its environment there is evidence for differentialdepositionof recently available calcium in the exoskeleton. Further, intermoltand early premolt animals maintained in Ca⁴⁵-labelled waterfor 15 days concentrate it 4 and 3—fold in the exoskeletonand stomach, respectively. However, removal of a molt-inhibitingfactor through ablation of eyestalks results in a 20 and 40—foldincrease in incorporation inthese same sites relative to environmentalconcentrations. Treatment with mammalian parathyroid extract mobilizes bothexoskeletal and gastric calciumand leads to a rise in bloodcalcium. However, there is little or no effect on levels ofexoskeletal citric acid. Further, citric acid is higher in thecrayfish carapace during stage C, the period of mineralization,than in stage D, the period of demineralization. There are both similarities and differences between the effectsof crustacean and mammalianregulating factors with respect tothe direction and extent of mineralization. Biochemical studiesshould elucidate the mechanisms regulated by these hormones.     

Effects of Calcium Ions and of Calcium Channel Blockers on Galvanotaxis of Chlamydomonas reinhardtii

The effects of calcium ions and of the calcium channel blockers verapamil, diltiazem and nifedipine on galvanotaxis in Chlamydomonas have been investigated using a fully automated and computerized population system. Galvanotaxis is a function of the voltage applied to the cell population. However, the galvanotactic orientation also depends on the external calcium concentration. In a calcium-deprived nutrient medium which still contains 6 × 10^?7M calcium, galvanotactic orientation is about 20% of orientation at optimal calcium concentration of 10^?4 M at 9 V. The higher the external calcium concentration is, the lower is the voltage necessary for optimal galvanotactic orientation. The calcium channel blockers diltiazem and nifedipine likewise inhibit galvanotaxis of Chlamydomonas very specifically without impairing motility. Verapamil is effective, but also inhibits motility by causing detachment or shortening of the flagella. Nevertheless, inhibition of galvanotaxis by verapamil is not the only result of decreased motility, because the galvanotactic orientation is impaired to a greater extent than motility. The effectiveness of the three blockers tested in inhibiting galvanotaxis depends on the concentration and on the voltage applied. At 10^?5 M, verapamil causes maximal inhibition of galvanotaxis at 9 V. At increasing concentrations up to 10^?4 M, diltiazem inhibits galvanotaxis more strongly than the other blockers. If the voltage is varied at a constant blocker concentration of 2 × 10^?5 M, nifedipine causes maximal inhibition at 3 V–6 V, diltiazem at 9 V and verapamil above 12 V.     

Anticarcinogenic effects of halofuginone on lung‐derived cancer cells

Malignant mesothelioma is a rare but aggressive form of malignancy, which is difficult to diagnose and is resistant to current chemotherapeutic treatment options. Molecular techniques have been used to investigate the mechanisms of action and the beneficial therapeutic effects of halofuginone (HF) in several cancers but not malignant mesotheliomas. In this study, the antiproliferative and apoptotic effects of HF were investigated through its ability to deregulate EGFR downstream signalling cascade proteins in the pathologically aggressive malignant mesothelioma and non‐small‐cell lung cancer cells. We showed that administration of HF at nanomolar concentrations induced a dose‐dependent reduction in the viability of cancer cells, made cell cycle arrest, inhibited proliferation of cancer cells via STAT3 and ERK1/2 pathways and triggered the apoptotic cascade via p38MAPK. We demonstrated that the apoptotic cell death mechanism was mediated by enhanced activation of caspase‐3 and concomitant PARP cleavage, downregulation of Bcl‐2 and upregulation of Bax in both malignant mesothelioma and lung cancer cells. In particular, we demonstrated that cancer cells were more sensitive to HF treatment than normal mesothelial cells. Taken together, this study suggests that HF exerts its anticancer effects in lung‐derived cancers by targeting signal transduction pathways mainly through deregulation of ERK1/2, STAT3 and p38MAPK to reduce cancer cell viability, induce cell cycle arrest and apoptotic cell death. Thus, HF might be considered as a potential agent against malignant mesothelioma and/or lung cancer cells.     

Anticarcinogenic Reactivity of Copper-Dischiffbases with Superoxide Dismutase-Like Activity

CuPu(Py)₂ and CuPu(Im)₂, two novel dischiffbase coordinated low Mr active centre analogues of Cu_{2, Zn2} superoxide dismutase, were shown to effectively catalyze the production of hydroxyl radicals in the presence and absence of TPA activated polymorphonuclear leukocytes. These stable copper chelates exhibited a pronounced anticarcinogenic reactivity in male Sprague Dawley rats implanted with Walker 256 carcinosarcoma cells. When four doses of 5°mol/kg CuPu(Py)₂ and CuPu(Im)₂, respectively, were administered intratumorally, reduction in tumor size, delay of metastasis and a significant increase in survival of the hosts were observed, resulting in 75% of total remissions. 60% of the animals recovered totally from the carcinosarcoma, when CuPu(Py)₂ was applicated intravenously.     

Anticarcinogenic Reactivity of Copper-Dischiffbases with Superoxide Dismutase-Like Activity

CuPu(Py)₂ and CuPu(Im)₂, two novel dischiffbase coordinated low Mr active centre analogues of Cu_{2, Zn2} superoxide dismutase, were shown to effectively catalyze the production of hydroxyl radicals in the presence and absence of TPA activated polymorphonuclear leukocytes. These stable copper chelates exhibited a pronounced anticarcinogenic reactivity in male Sprague Dawley rats implanted with Walker 256 carcinosarcoma cells. When four doses of 5°mol/kg CuPu(Py)₂ and CuPu(Im)₂, respectively, were administered intratumorally, reduction in tumor size, delay of metastasis and a significant increase in survival of the hosts were observed, resulting in 75% of total remissions. 60% of the animals recovered totally from the carcinosarcoma, when CuPu(Py)₂ was applicated intravenously.     

环境钙对鲤鱼血浆钙含量及斯坦尼氏小体结构的影响

将鲤鱼①饲养在富钙淡水(天然淡水中添加0．5％CaCl2)中,②在富钙淡水饲养8d后分别腹腔注射维生素D3(VD3,5000U／100g体重,以提高细胞外钙浓度)和鲤鱼斯坦尼氏小体(CS)匀浆液(0．5mg/100g体重,以降低细胞外钙浓度),③富钙淡水饲养28d后转入天然淡水。单纯的高钙环境能够诱导鲤鱼出现高血钙;注射VD3和CS匀浆液则分别引起血钙水平升高和降低。血钙升高时,CS囊泡萎缩,分泌细胞的核增大、粗面内质网增多、细胞脱颗粒化,提示CS分泌细胞在Ca^2 的作用下合成与分泌活动旺盛,但过度刺激将导致分泌细胞的衰竭与破裂;当血钙降低时,上述症状有所缓解,分泌细胞内的分泌颗粒积累,提示CS分泌细胞的分泌活动减弱,合成活动正常;从高钙环境转入天然淡水后,CS分泌细胞的结构与功能亦可逐渐恢复正常。鲤鱼CS的状态受细胞外和体外钙水平变动的影响,而且这种影响是可逆的。