Tissue Calcium Research Articles

Iron and Folic Acid Supplementation Affects Mineral Status in Female Rats with a Deficiency of These Micronutrients

Supplementation with iron and folic acid is widely recommended in women of childbearing age and during pregnancy; however, the effect of such supplementation on mineral status is not well-known. The aim of this study was to determine the effects of oral iron and folic acid, administered together and separately, on copper, zinc, calcium, and magnesium concentrations in the tissues of rats with a deficiency of both these micronutrients. The experiment was performed on 8-week-old female Wistar rats. In the first stage of the experiment, the animals were randomly assigned to a control group of rats fed the standard diet (AIN-93 M), and to a study group of rats fed a diet deficient in iron and folate. The study group was then randomly divided to four groups: group D was fed a deficit diet, group FE was fed a deficit diet with iron gluconate, the FOL group was fed a deficit diet with folate acid, and the FEFOL group was fed a deficit diet with iron gluconate and folate acid. After 2, 10, and 21 days of the intervention, ten animals from each group were killed. Mineral concentrations were assayed in the liver, spleen, pancreas, heart, and kidneys using atomic absorption spectrometry. Statistical analysis was performed using Statistica 12.0 with the ANOVA test (p < 0.05). It was found that separate supplementation with iron and folic acid significantly decreased copper concentrations in tissues. The deficit in iron and folic acid decreased, and their simultaneous supplementation increased calcium content in the organs. Separate and simultaneous supplementation decreased magnesium status in deficient rats. In conclusion, iron and folic acid, supplemented separately or simultaneously, affect the copper, calcium, and magnesium level in tissues.

Conventional and Transgenic Soybeans: Physiological and Nutritional Differences in Productivity under Sulfur Fertilization

ABSTRACT Increasingly intensive cropping systems and the introduction of new genetically modified cultivars can lead to a distinct demand for nutrients in soybean crops. This study aims to evaluate the response of two soybean cultivars [with and without tolerance to glyphosate (RR)] to sulfur (S) rates. The experimental design was completely randomized in a 6 × 2 factorial arrangement with four replicates. The treatments consisted of six S rates (0, 10, 20, 40, 80, and 120 mg kg−1) and two soybean cultivars from the same breeding program (BRS 317 and BRS 360RR). The increase in S rates resulted in the herbicide sensitive cultivar (BRS 317) having the best physiological indexes (water use efficiency and chlorophyll content) when compared with those of herbicide tolerant cultivar (BRS 360RR), and an increased grain yield (GY), shoot dry weight yield (SDWY), calcium (Ca), boron (B), NO2-N, and iron (Fe) concentration in the leaf tissues. This indicates that the RR gene can cause significant differences in S uptake responses.

A fluorogenic cyclic peptide for imaging and quantification of drug-induced apoptosis

Programmed cell death or apoptosis is a central biological process that is dysregulated in many diseases, including inflammatory conditions and cancer. The detection and quantification of apoptotic cells in vivo is hampered by the need for fixatives or washing steps for non-fluorogenic reagents, and by the low levels of free calcium in diseased tissues that restrict the use of annexins. In this manuscript, we report the rational design of a highly stable fluorogenic peptide (termed Apo-15) that selectively stains apoptotic cells in vitro and in vivo in a calcium-independent manner and under wash-free conditions. Furthermore, using a combination of chemical and biophysical methods, we identify phosphatidylserine as a molecular target of Apo-15. We demonstrate that Apo-15 can be used for the quantification and imaging of drug-induced apoptosis in preclinical mouse models, thus creating opportunities for assessing the in vivo efficacy of anti-inflammatory and anti-cancer therapeutics.

Usefulness of Thrombin Generation.

Thrombin generation (TG) is a global coagulation procedure meant to continuously monitor thrombin formation and decay upon exposure of platelet-poor plasma to exogenous triggers such as tissue factor, phospholipids, and calcium chloride. The procedure can also be performed in platelet-rich plasma by omitting exogenous phospholipids. TG is thought to mimic more than other coagulation procedures the process that occurs in vivo. Over the years, TG has been used to investigate coagulation mechanisms, which were not completely understood, or to investigate hyper- or hypocoagulability in clinical conditions known to be at increased risk of thrombosis or hemorrhage, respectively. More recently, TG has been employed as a laboratory tool to assess the risk of recurrent venous thromboembolism or to assess the risk of arterial thromboembolism in specific clinical settings or in the general population. The article reviews the value and limitations of TG.

Can tissue elemental analysis be used to differentiate sarcoidosis and tuberculous lymphadenitis?

Sarcoidosis and tuberculous lymphadenitis are granulomatous inflammatory diseases. Differentiating lymph node involvement in these two diseases can be challenging. This study evaluated whether elemental analysis of tissue samples could facilitate the differentiation of these histopathologically and clinically similar diseases. A total of 152 tissue samples were included: 57 caseating granulomatous inflammation, 58 non-caseating granulomatous inflammation, and 37 reactive lymph node specimens. The tissue samples were analyzed for calcium, magnesium, iron, copper, zinc, chrome, molybdenum, nickel and selenium with inductively coupled plasma-optical emission spectroscopy (ICP-OES). Comparison of element levels in the three groups revealed that caseating granulomatous inflammation had higher calcium content (662.6 ± 4.6 ppm, p< 0.001) and lower iron content (48.7 ± 83 ppm, p< 0.001) compared to non-caseating granulomatous inflammation. Compared to reactive lymph tissue, caseating granulomatous inflammation had higher calcium and lower iron and magnesium content while non-caseating granulomatous inflammation had higher levels of iron and lower magnesium; however, these differences were not statistically significant. In caseating granulomatous inflammation, a calcium cut-off value of 207 ppm yielded 85% specificity and 63% sensitivity. For iron, a cut-off value of 51 ppm had 74% specificity and 58% sensitivity. High calcium and low iron levels in lymph tissue may be suggestive of caseating granulomatous inflammation and tuberculosis. In cases where differentiating between sarcoidosis and tuberculous lymphadenitis is difficult, performing tissue elemental analysis may provide additional supportive evidence for differential diagnosis.

Paediatric osteoporosis: features of skeletal mineralization in children, prevention, and treatment

The prevention and treatment of osteoporosis in children is an urgent problem of pediatrics and pediatric endocrinology. This is due to the need for preventive measures for the formation of adequate bone mass and bone strength, the duration of rehabilitation during the development of the disease and the occurrence of fractures, as well as the requirements for optimizing the treatment of this pathology in childhood.Bone tissue is a dynamic and constantly updated system. The main regulators of osteogenesis include trace elements (calcium, phosphorus, zinc, fluorine, etc.), vitamin D, hormones (parathyroid hormone, calcitonin, growth hormone, corticosteroids, etc.), growth factors (IRF-1, IRF-2, ERF, FRF, etc.), local bone-resorbing factors (IL-1, IL-6, TNF, PGE, etc.) and mechanical stress. Remodeling processes continue throughout life, but are most intense until 18 years of age.The most important risk factors for developing osteoporosis are a history of prematurity and / or IUGR, rickets, low intake of protein, calcium and vitamin D, weight deficiency, periods of active growth, low physical activity, endocrine diseases (diabetes mellitus, growth hormone deficiency, hypogonadism, etc.), somatic pathology (chronic diseases of the kidneys, liver, malabsorption), taking certain medications (GCS, anticonvulsants, cytostatics, etc.). Clinical manifestations of osteoporosis in children include signs of tissue calcium deficiency (trophic changes in the nails, hair and tooth enamel, paresthesia, pain in the calf muscles) and a decrease in bone mineralization (impaired posture, slowed growth, pain in the spine and large joints, pathological low-energy fractures) Measures to prevent osteoporosis in childhood include regular physical activity, a balanced diet, adequate insolation, and the identification and treatment of somatic pathology. Additional pharmacological correction is carried out in risk groups – the recommended course use of preparations containing vitamin D, calcium and other osteotropic minerals.The treatment of osteoporosis in children and adolescents is actively debated. A mandatory component of therapy are calcium salts in combination with vitamin D. The possibility of using bisphosphonates is being examined, the effect of monoclonal antibodies (denosumab), somatropin and anabolic steroids on bone metabolism is being studied. In addition, non-pharmacological methods are prescribed - orthopedic correction for fractures, exercise therapy, gymnastics, therapeutic swimming and massage.

Salinity Tolerance of Select Wildflower Species in a Hydroponic Setting

Water quality and quantity are critical issues in the Southwest United States and many other locations in the world. Use of reclaimed water for landscape irrigation can conserve potable water significantly and possibly reduce fertilizer application. A potential concern of using alternative water sources is elevated salt levels, which can have adverse effects on plant growth and aesthetic appearance. Most Texas native wildflowers are known to be hardy and easy to maintain, and are drought tolerant after establishment. In addition, native wildflowers provide wildlife habitat and support native pollinators. However, little information is available on salinity tolerance of many Texas native wildflower species. In this study, two separate hydroponic experiments were conducted to determine salt tolerance of three Texas native wildflower species: Gaura villosa Torr. (wooly gaura), Xanthisma texanum DC. (Texas sleepy daisy), and Ipomopsis rubra (L.) Wherry (standing cypress). Species were suspended in a hydroponic setting using a randomized complete block design with a control [municipal reverse-osmosis (RO) water with a nutrition solution at an electrical conductivity (EC) of 3.0 dS·m–1] and three salinity treatments: 5.0, 7.0, and 11.0 dS·m–1 EC. Sixty days after salinity treatments were initiated, percent survival, visual rating, fresh weight, and length measurements were recorded on root and shoot tissue. To determine tissue percentage sodium (Na+), calcium (Ca2+), and chloride (Cl–), shoot and root tissues were dried and ground for tissue analysis. At the end of each experiment, total percent survival for X. texanum, G. villosa, and I. rubra were 100%, 94%, and 76%, respectively, with the greatest mortality rate at the highest salinity treatment. Shoot dry weight and plant growth index (PGI) decreased in all three species as salinity of irrigation water increased. Visual qualities of all species were mainly compromised at the highest salinity level. Ion concentrations in root and shoot tissues were affected by salinity levels and varied among species. Different mechanisms of salt tolerance (ion exclusion, salt excretion, and tissue tolerance to high concentrations of Na+ or Cl–) have been observed among wildflower species, and results indicate different salt tolerance mechanisms were exhibited by each trial species. In addition, results indicate I. rubra can be identified as moderately salt tolerant (EC up to 7.0 dS·m–1), whereas, X. texanum and G. villosa can be classified as salt tolerant (EC up to 11.0 dS·m–1). Results from this study suggest great potential of these native Texas wildflowers in landscapes using limited-quality irrigation water or landscapes with soil salinity concerns.

Protective Effects of Chrysin Against Oxidative Stress and Inflammation Induced by Lead Acetate in Rat Kidneys: a Biochemical and Histopathological Approach

In this study, the protective effects of chrysin (CR) on lead acetate (PbAc)-induced renal toxicity in Sprague-Dawley rats were investigated with biochemical, histopathological, and immunohistochemical methods. In the study, rats were given orally at 30 mg/kg/body weight (BW) PbAc after CR of 25 and 50 mg/kg/BW was administered to them orally (a total of 7 administrations for 7 days). The results showed that CR reduced urea and creatinine levels by alleviating PbAc-induced kidney damage. It was determined that CR decreases PbAc-induced lipid peroxidation due to its antioxidant properties and increases catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx) activities, and glutathione (GSH) levels. It was also detected that CR protects DNA from the toxic effects of PbAc and reduces 8-hydroxy-2'-deoxyguanosine (8-OHdG) levels. Biochemical and immunohistochemical findings demonstrated that CR had anti-inflammatory and antiapoptotic effects and reduced nuclear factor kappa-B (NF-κB), interleukin-33 (IL-33), prostaglandin-E2 (PGE-2), tumor necrosis factor-α (TNF-α), p53 levels, and the activities of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS), which were increased with PbAc administration. Moreover, CR was found to increase the levels of aquaporin-1 (AQP-1) and nephrine in PbAc-induced kidney tissue. CR decreased the contents of lead (Pb), zinc (Zn), iron (Fe), sodium (Na), and copper (Cu) and increased those of potassium (K) calcium (Ca) in renal tissue. These results indicated that CR considerably alleviates kidney toxicity caused by PbAc.

The Inhibitory Ability of Orobanche Extract on Calcium Oxalate Lithiasis in Male Albino Rats

The present study aimed to evaluate the inhibitory effect of Orobanche extract in ethylene glycol induced nephrolithiasis. Thirty male albino rats were divided into five groups each group contains 6 animals, group (1) control group, group (2) animals were supplied with 0.75% ethylene glycol in drinking water, group (3) animals were administrated Orobanche extract 3g/kg orally, group (4) animals were administrated Cystone 500 mg/kg in addition to 0.75% ethylene glycol, group (5) animals were administrated Orobanche extract 3g/kg orally in addition to 0.75% ethylene glycol the experiment continued for 28 days. Serum and the kidney homogenates were analyzed for various biochemical parameters and urine was examined microscopically for crystals. Orobanche treatment group and Cystone treatment group significantly decreased phosphorus, Calcium and Oxalate in kidney tissue of nephrolithiasis rats and significantly decreased kidney and liver marker in serum of nephrolithiasis rats. Conclusion this result revealed that Orobanche extract could be a potential candidate for phytotherapy against nephrolithiasis.

Morphology and serum and bone tissue calcium and magnesium concentrations in the bones of male rats chronically treated with letrozole, a nonsteroidal cytochrome P450 aromatase inhibitor

ABSTRACT Purpose/Aim of the study: The role of estrogen (E) in the regulation of bone turnover in women is well established, though the contributions of E versus testosterone (T) in the control of bone turnover in men are poorly understood. The aim of this study was to examine the association between chronic treatment with letrozole, a nonsteroidal inhibitor blocking the aromatase activity and thus the conversion of androgens into estrogens, and cortical bone morphology in the femur and humerus of male adult rats. Materials and Methods: Adult male rats were treated with letrozole for 6 months and the body and femur weight, morphology, collagen structure, blood serum, and bone tissue concentrations of calcium and magnesium were examined. Results: Long-term aromatase inhibition resulted in a decrease in femur mass, a wavelike arrangement of bone and lamellae with an altered organization of collagen in compact bone, a increased concentration of calcium in blood serum, and no change in calcium bone tissue concentration, magnesium serum, or bone tissue concentration. MicroCT study of the humerus revealed significant decreases of whole bone tissue volume, cortical bone thickness, cortical bone volume, and external cortical bone thickness with letrozole treatment. Conclusion: Chronic treatment with letrozole affected cortical bone structure and produced histomorphological changes in male rat bone similar to that observed in the aging processes.

The impact of humic acid additives on mineral metabolism of rabbits in the postnatal period of ontogenesis

Humic acids are characterized by chelating properties due to which they are able to influence the mineral metabolism in animals. Rabbits have species-specific characteristics of mineral metabolism, which distinguishes them from most species of mammals. For the experiment, 16 rabbits of 45-day-old of Hyplus breed were divided into the experimental and control group of 8 rabbits in each. The animals were 15 days in the preparatory period. The rabbits of the experimental group received humic acid additive individually with water in the amount of 5 mg/kg, the rabbits of the control group received pure water in the same amount. The ability of humic acid additive to increase the content of calcium, ionized calcium, iron, to correct the content of copper and zinc, and also to increase the activity of alkaline phosphatase in the blood serum of rabbits of the experimental group has been proved. We observed the early effect of the humic acid additive on increasing the content of iron and calcium ionized and a later effect on the activity of alkaline phosphatase and an increase in the content of copper in the blood serum. The influence of humic acid additive on the distribution of calcium, phosphorus, manganese, copper and zinc in bone tissue is determined. According to a histological study, there was a significant increase in the number of layers of osteons and osteoblasts in the bone tissue of the femur, an increase in the number of osteons and osteoblasts in the bone tissue of the sternum and an increase in the number of columns of chondrocytes and an increase in the number of chondrocytes in the column in the cartilaginous tissue of the sternum. The results of a histological study of bone tissue, together with an increase in structural macroelements in it and redistribution of osteotropic trace elements in the blood and bone tissue suggest the possibility of using a humic acid additive to intensify the growth and mineralization of bone tissue, which can improve the state of bone tissue of rabbits in the postnatal period ontogenesis.

The impact of humic acid additives on mineral metabolism of rabbits in the postnatal period of ontogenesis

Humic acids are characterized by chelating properties due to which they are able to influence the mineral metabolism in animals. Rabbits have species-specific characteristics of mineral metabolism, which distinguishes them from most species of mammals. For the experiment, 16 rabbits of 45-day-old of Hyplus breed were divided into the experimental and control group of 8 rabbits in each. The animals were 15 days in the preparatory period. The rabbits of the experimental group received humic acid additive individually with water in the amount of 5 mg/kg, the rabbits of the control group received pure water in the same amount. The ability of humic acid additive to increase the content of calcium, ionized calcium, iron, to correct the content of copper and zinc, and also to increase the activity of alkaline phosphatase in the blood serum of rabbits of the experimental group has been proved. We observed the early effect of the humic acid additive on increasing the content of iron and calcium ionized and a later effect on the activity of alkaline phosphatase and an increase in the content of copper in the blood serum. The influence of humic acid additive on the distribution of calcium, phosphorus, manganese, copper and zinc in bone tissue is determined. According to a histological study, there was a significant increase in the number of layers of osteons and osteoblasts in the bone tissue of the femur, an increase in the number of osteons and osteoblasts in the bone tissue of the sternum and an increase in the number of columns of chondrocytes and an increase in the number of chondrocytes in the column in the cartilaginous tissue of the sternum. The results of a histological study of bone tissue, together with an increase in structural macroelements in it and redistribution of osteotropic trace elements in the blood and bone tissue suggest the possibility of using a humic acid additive to intensify the growth and mineralization of bone tissue, which can improve the state of bone tissue of rabbits in the postnatal period ontogenesis.

Evocalcet prevents ectopic calcification and parathyroid hyperplasia in rats with secondary hyperparathyroidism.

Elevated parathyroid hormone (PTH) levels in secondary hyperparathyroidism (SHPT) lead to vascular calcification, which is associated with cardiovascular events and mortality. Increased PTH production is caused by the excessive proliferation of parathyroid gland cells, which is accelerated by abnormal mineral homeostasis. Evocalcet, an oral calcimimetic agent, inhibits the secretion of PTH from parathyroid gland cells and has been used for the management of SHPT in dialysis patients. We observed the effects of evocalcet on ectopic calcification and parathyroid hyperplasia using chronic kidney disease (CKD) rats with SHPT. CKD rats with SHPT induced by adenine received evocalcet orally for 5 weeks. The calcium and inorganic phosphorus content in the aorta, heart and kidney was measured. Ectopic calcified tissues were also assessed histologically. To observe the effects on the proliferation of parathyroid gland cells, parathyroid glands were histologically assessed in CKD rats with SHPT induced by 5/6 nephrectomy (Nx) after receiving evocalcet orally for 4 weeks. Evocalcet prevented the increase in calcium and inorganic phosphorus content in the ectopic tissues and suppressed calcification of the aorta, heart and kidney in CKD rats with SHPT by reducing the serum PTH and calcium levels. Evocalcet suppressed the parathyroid gland cell proliferation and reduced the sizes of parathyroid cells in CKD rats with SHPT. These findings suggest that evocalcet would prevent ectopic calcification and suppress parathyroid hyperplasia in patients with SHPT.

Regulation of calcium dynamics and propagation velocity by tissue microstructure in engineered strands of cardiac tissue.

Disruptions to cardiac tissue microstructure are common in diseased or injured myocardium and are known substrates for arrhythmias. However, we have a relatively coarse understanding of the relationships between myocardial tissue microstructure, propagation velocity and calcium cycling, due largely to the limitations of conventional experimental tools. To address this, we used microcontact printing to engineer strands of cardiac tissue with eight different widths, quantified several structural and functional parameters and established correlation coefficients. As strand width increased, actin alignment, nuclei density, sarcomere index and cell aspect ratio decreased with unique trends. The propagation velocity of calcium waves decreased and the rise time of calcium transients increased with increasing strand width. The decay time constant of calcium transients decreased and then slightly increased with increasing strand width. Based on correlation coefficients, actin alignment was the strongest predictor of propagation velocity and calcium transient rise time. Sarcomere index and cell aspect ratio were also strongly correlated with propagation velocity. Actin alignment, sarcomere index and cell aspect ratio were all weak predictors of the calcium transient decay time constant. We also measured the expression of several genes relevant to propagation and calcium cycling and found higher expression of the genes that encode for connexin 43 (Cx43) and a subunit of L-type calcium channels in thin strands compared to isotropic tissues. Together, these results suggest that thinner strands have higher values of propagation velocity and calcium transient rise time due to a combination of favorable tissue microstructure and enhanced expression of genes for Cx43 and L-type calcium channels. These data are important for defining how microstructural features regulate intercellular and intracellular calcium handling, which is needed to understand mechanisms of propagation in physiological situations and arrhythmogenesis in pathological situations.

Microvasculopathy and soft tissue calcification in mice are governed by fetuin-A, magnesium and pyrophosphate.

Calcifications can disrupt organ function in the cardiovascular system and the kidney, and are particularly common in patients with chronic kidney disease (CKD). Fetuin-A deficient mice maintained against the genetic background DBA/2 exhibit particularly severe soft tissue calcifications, while fetuin-A deficient C57BL/6 mice remain healthy. We employed molecular genetic analysis to identify risk factors of calcification in fetuin-A deficient mice. We sought to identify pharmaceutical therapeutic targets that could be influenced by dietary of parenteral supplementation. We studied the progeny of an intercross of fetuin-A deficient DBA/2 and C57BL/6 mice to identify candidate risk genes involved in calcification. We determined that a hypomorphic mutation of the Abcc6 gene, a liver ATP transporter supplying systemic pyrophosphate, and failure to regulate the Trpm6 magnesium transporter in kidney were associated with severity of calcification. Calcification prone fetuin-A deficient mice were alternatively treated with parenteral administration of fetuin-A dietary magnesium supplementation, phosphate restriction, or by or parenteral pyrophosphate. All treatments markedly reduced soft tissue calcification, demonstrated by computed tomography, histology and tissue calcium measurement. We show that pathological ectopic calcification in fetuin-A deficient DBA/2 mice is caused by a compound deficiency of three major extracellular and systemic inhibitors of calcification, namely fetuin-A, magnesium, and pyrophosphate. All three of these are individually known to contribute to stabilize protein-mineral complexes and thus inhibit mineral precipitation from extracellular fluid. We show for the first time a compound triple deficiency that can be treated by simple dietary or parenteral supplementation. This is of special importance in patients with advanced CKD, who commonly exhibit reduced serum fetuin-A, magnesium and pyrophosphate levels.

Fractionating of Calcium in Tuber and Leaf Tissues Explains the Calcium Deficiency Symptoms in Potato Plant Overexpressing CAX1.

Consistent with reports on other plants we recently reported that a potato transgenic line (AT010901) overexpressing sCAX1 show classic symptoms of calcium deficiency shoot tip injury, leaf curling, leaf margin necrosis and tuber internal defects such as hollow heart and brown spots. The present study was undertaken to quantify calcium in various fraction of leaf and tuber tissues of this transgenic and wild type potato clones to understand the development of these deficiency symptoms at normal calcium nutrition (1mM) and its mitigation at higher calcium nutrition (10mM). Plants were grown in controlled environment growth chamber and watered with balanced nutrient solution containing either 1 or 10 mM calcium. The plants overexpressing sCAX1 showed calcium deficiency symptoms while sequestering calcium in the vacuole as calcium oxalate crystals. Various fractions of calcium were qualified in the young and mature leaves as well as tuber tissue. A reduced concentration of water soluble fraction of calcium was most important factor related to the development of calcium deficiency symptoms in the line overexpressing sCAX1. Furthermore, an increase in this fraction appear to explain the alleviation of the deficiency symptoms in these transgenic plants.Ours is the first study to document the significance of water-soluble calcium in the development of calcium-deficiency symptoms in the potato transgenic lines overexpressing sCAX1. Furthermore, our result demonstrates that an increase in this fraction plays a significant role in the alleviation of calcium deficiency symptoms when calcium concentration in the nutrient media is increased. These results provide important insight on the role of sCAX1 in the calcium homeostasis.

X-ray fluorescence elemental mapping of roots, stems and leaves of the nickel hyperaccumulators Rinorea cf. bengalensis and Rinorea cf. javanica (Violaceae) from Sabah (Malaysia), Borneo

There are major knowledge gaps in understanding the translocation leading from nickel uptake in the root to accumulation in other tissues in tropical nickel hyperaccumulator plant species. This study focuses on two species, Rinorea cf. bengalensis and Rinorea cf. javanica and aims to elucidate the similarities and differences in the distribution of nickel and physiologically relevant elements (potassium, calcium, manganese and zinc) in various organs and tissues. High-resolution X-ray fluorescence microscopy (XFM) of frozen-hydrated and fresh-hydrated tissue samples and nuclear microprobe (micro-PIXE) analysis of freeze-dried samples were used to provide insights into the in situ elemental distribution in these plant species. This study has shown that the distribution pattern of nickel hyperaccumulation is typified by very high levels of accumulation in the phloem bundles of roots and stems. In the leaves, nickel is preferentially located in epidermal cell region, whereas manganese is located mainly in the lower epidermis and zinc in the upper epidermis and palisade mesophyll. The abundant formation of calcium-oxalate crystals, lining the collenchyma, is a prominent feature of both Rinorea cf. bengalensis and Rinorea cf. javanica. Future investigations on Rinorea cf. bengalensis and Rinorea cf. javanica should focus on unravelling the mechanisms of nickel uptake in the root, specifically targeting the identification of nickelspecific membrane transporters.

Стан кісткової тканини курчат-бройлерів за використання у раціоні кормових добавок, виготовлених із морських гідробіонтів

The purpose of the work was to determine the content of Calcium and Phosphorus in bone tissue, as well as the tibia strength index of broiler chickens under using two feed additives made from the waste of primary processing of marine hydrobionts in the diet. The experiment was carried out on “Ross 308” cross broilers, which were divided into 5 groups: 1 control and 4 experimental. Each additive was fed in two ways: by replacing 7% of the diet weight and adding 7% to it. Chickens of the control group were fed only the basic diet (BD), balanced by basic nutrients. Chickens of experimental groups from 20 to 42 days old were additionally fed the feed additives. Chickens D-1 and D-2 groups consumed a mineral feed additive (MFA), D-3 and D-4 groups received a protein-mineral feed additive (PMFA). Moreover, broilers of D-1 and D-3 groups by feed weight received 93% of BD and 7% of MFA and PMFA respectively, D-2 and D-4 groups in addition to BD (100%) received 7% respectively of MFA and PMFA. MFA is milled leaves of the Black Sea Mussel (Mytilus galloprovincialis) and seawater. PMFA contained milled leaves of large and the body of small mussels, waste from the primary processing of red algae Phyllophora nervosa and seawater. As a material for research was considered the chickens’ tibia of of 42 days old. The concentration of total Calcium and inorganic Phosphorus was determined in the middle part of the tibia using a UV mini-1240 SHIMADZU spectrophotometer (Japan) and a set of reagents NPP Filisit-Diagnostika (Ukraine). The bone cross-sectional area was determined by scanning prior to testing with processing in the Compass-3D graphics program. The mechanical parameters of the bones were determined on an FM-1000 testing machine under the action of a double-support bend. The use of feed additives contributed to an increase in the content of Calcium and Phosphorus in the bone tissue of broiler chickens and, accordingly, the strength of the tibia. The Phosphorus content increase took place to a greater extent than Calcium, which caused a decrease in the Calcium-Phosphorus ratio. The content of Calcium and Phosphorus in the bone tissue, as well as the strength of the tibia of the chickens increased due to diet enrichment with feed additives, and not its partial replacement.

Spring triticale forage responses to seeding rate and nitrogen application

AbstractGrowing spring forages could reduce soil erosion and increase cropping system profitability compared with summer fallow in a wheat (Triticum aestivum L.)–sorghum (Sorghum bicolor L. Moench ssp. Bicolor)–fallow system. Field experiments were conducted over four site‐years to evaluate seeding rate and nitrogen (N) application effects on spring triticale forage production. Seeding rates (60, 80, and 100 kg ha−1) were assigned to main plot treatments, and N rates (0, 11, 34, 56, and 78 kg N ha−1) were the subplots. Seeding rate had no effect on forage dry matter (DM), crude protein (CP), or mineral nutrients. However, increasing seeding rate increased lignin, acid detergent fiber (ADF), and neutral detergent fiber (NDF) concentrations while decreasing in vitro dry matter digestibility (IVDMD) and total digestible nutrients. Applying N increased forage DM, with optimum yield occurring at 56 kg N ha−1. The application of N fertilizer decreased ADF and NDF concentrations but increased CP, IVDMD, and total digestible nutrients. Similarly, tissue calcium, phosphorus, and potassium concentrations increased with N fertilizer application. Based on our findings, 56 kg N ha−1 was adequate for spring triticale and seeding triticale forage at a lower rate of 60 kg ha−1 had no detrimental effect on forage yield when harvested as hay.

The effect of amoxicillin on dental enamel development in vivo

The exposure to amoxicillin has been associated with molar incisor hypomineralization. This study aimed to determine if amoxicillin disturbs the enamel mineralization in in vivo experiments. Fifteen pregnant rats were randomly assigned into three groups to received daily phosphatase-buffered saline or amoxicillin as either 100 or 500 mg/kg. Mice received treatment from day 13 of pregnancy to day 40 postnatal. After birth, the offsprings from each litter continued to receive the same treatment according to their respective group. Calcium (Ca) and phosphorus (P) content in the dental hard tissues were analyzed from 60 upper first molars and 60 upper incisors by the complexometric titration method and colorimetric analysis using a spectrophotometer at 680 nm, respectively. Lower incisors were analyzed by X-ray microtomography, it was measured the electron density of lingual and buccal enamel, and the enamel and dentin thickness. Differences in Ca and P content and electron density among the groups were analyzed by one-way ANOVA. There was no significant difference on enamel electron density and thickness among the groups (p > 0.05). However, in incisors, the higher dose of amoxicillin decreased markedly the electron density in some rats. There were no statistically significant differences in Ca (p = 0.180) or P content (p = 0.054), although the higher dose of amoxicillin could affect the enamel in some animals. The amoxicillin did not significantly alter the enamel mineralization and thickness in rats.