Ceruloplasmin is a major ferroxidase in sera, and people with hypoceruloplasminemia might have iron load complications. We used histochemical and X-ray microanalysis techniques on pretreatment liver specimens from five patients with Wilson's disease. Serum levels of ceruloplasmin and ferritin were determined before chelation therapy. All the patients showed hypoceruloplasminemia ranging from 15 to 160 mg/L. Histo-chemical copper was positively stained in four of the five patients. Copper deposits were clearly observed in the hepatocyte lysosomes of one patient. High serum ferritin levels ranging from 188 to 400 ng/mL were found. Histochemical iron was positive in the livers of two patients. Iron-specific X-rays were positive in the hepatocyte lysosomes of all the patients. These findings suggest that compound load of copper and iron is a fundamental finding in the liver of Wilson's disease with hypoceruloplasminemia. Copper cytotoxicity is a primary target for chelation therapy, and iron load, if hepatotoxic, should be treated by phlebotomy. J. Trace Elem. Exp. 17:65–73, 2004. © 2004 Wiley-Liss, Inc.
In this pilot study, 13 dogs of various breeds and ages with a variety of advanced tumors were treated with the antiangiogenic copper complexing drug tetrathiomolybdate. Dose escalations were performed to determine the safe and effective dose in dogs. The study was designed to last for a 6-month period for each dog entered. Dogs were examined weekly for the first 3 months and then every other week for the next 3 months. Complete blood count, serum biochemistry panel, and measurement of serum ceruloplasmin (Cp) content were conducted at each visit to monitor response to drug administration. Serum Cp was used as a surrogate marker of copper status. The owner reported toxicity at each visit. The dog was examined for physical abnormalities. Tumor measurements were completed every 2 weeks using direct measurements of the tumor with calipers in three planes or by direct measurements of the tumor using radiographic or ultrasonographic images in two planes. Tumor response was evaluated only after dogs achieved a 4-week reduction in their serum Cp-content. Tumor responses were defined as either disease stabilization or reduction in tumor volume. Nine dogs achieved Cp reduction. Of those dogs, five had no response to treatment, whereas four dogs had tumor responses after Cp reduction, characterized as either disease stabilization or reduction in tumor volume, for the remainder of the study period. There was only mild self-limiting toxicity with use of the drug.
This article summarizes recent studies and is a review of the literature on the fluoride compounds and their influence on bone mineral mass. It is well known that fluoride is an important element for mineralization of body tissues. The use of topical and systemic fluoride for oral health has resulted in major reduction in dental caries, but fluoride also plays a role in bone health. Dieticians and other health professionals are advised to recommend adequate use of systemic and topical fluorides, especially in children and adolescents. Population studies as well as earlier studies of individuals exposed to fluorides indicate that doses above 30–40 mg daily can result in fluorosis, characterized by increased fracture risk. Numerous clinical studies have demonstrated increased bone mineral density in subjects treated with appropriate doses of fluoride. However, the clinical interpretation of these studies has been a matter of debate. Therapeutically, fluoride seems to be useful when the agent is started in the early stages of osteoporosis, especially in patients with intact trabecular bone. It is well established that fluoride can act as an effective stimulator of bone formation by the osteoblasts. Bisphosphonates, which can also increase bone density, act by inhibiting bone resorption by osteoclasts. As a result of this, owing to the tight paracrine association characterizing skeletal metabolism, bone formation is slowed down by the antiresorptive agents. It is of particular interest that great benefit in the treatment of osteoporoses is seen when adequate osteoblast-stimulating doses of fluorides are given in combination with antiresorptive medication, but further research is needed to substantiate this promising concept. J. Trace Elem. Exp. Med. 17:83–92, 2004. © 2004 Wiley-Liss, Inc.
A quantitative elemental analysis of scalp hair of female breast cancer patients (n = 26) and female controls (n = 27) was used to Study to find out correlation and possible changes. between breast cancer and healthy controls. Atomic absorption spectrophotometer analysis of the quantitative method was used to determine copper, magnesium, and zinc levels. There were no differences in magnesium concentrations (P > 0.05) in breast cancer patients and healthy subjects. However, comparison of mean elemental contents of the breast cancer patients with controls showed a significant enhancement of copper (P < 0.05) but declining trends for zinc (P < 0.05) in breast cancer patients. The usefulness and significance of element status in cancer patients should be discussed more detailed in the light of the most these recent data. (C) 2004 Wiley-Liss, Inc.
Altered plasma levels of zinc, copper, and iron during pregnancy are known to have profound effects on pregnant women and their neonates. The status of these elements is not known in pregnant women in Jordan. During the three trimesters of pregnancy, blood specimens were collected from 186 healthy pregnant women aged 17-45 years and from cord blood of 92 of their neonates. The mean neonatal birth weight was 3.34 +/- 0.44 kg. Maternal and cord blood serum levels of zinc, copper, and iron were determined by atomic absorption spectrophotometry, and hemoglobin concentration was deter-mined by hematology cell counter. The results indicate significantly lower serum zinc levels and higher copper and iron levels in cord blood than in maternal blood. During the three trimesters of pregnancy, the serum levels of zinc and copper significantly decreased and increased, respectively, whereas the levels of serum iron were unchanged. Significant positive correlation was observed only between zinc levels of cord blood and birth weight. During third trimester, the mean serum levels of zinc and iron were significantly lower in anemic pregnant women (group I: Hb less than 11.0 g/ dL, n = 36) than that in nonanemic pregnant women (group-II: Hb > or = 11.0 g/dL, n = 56). There was no noticeable difference between group I and group 11 regarding cord blood parameters on one hand and neonatal birth weight on the other hand. Similar significant positive correlation was observed between serum zinc levels of cord blood and birth weight in both groups. These results indicate that Jordanian women during pregnancy follow a well-balanced and adequate diet regime.
The present experiment was designed to study the neurotoxic consequences of lead exposure on antioxidant enzymes like glutathione, glutathione peroxidase, super oxide dismustase, and catalase along with structural changes both in cerebrum as well as cerebellum. Lead was administrated orally in the doses of 50 mg/kg for a period of 8 weeks and study was performed at the end of exposure. Decrease in the concentration of all the antioxidant enzymes was observed, and after lead treatment, transverse sections of cerebrum showed degeneration of neurons. Disruption of normal arrangement of cell layers was seen. Cells were bigger in size with large vascular spaces around them. Lead treatment for 8 weeks also was enough to disrupt the normal arrangement of cellular layer of cerebellum. Large spaces in between purkinje cell layer and granular layer were seen. J. Trace Elem. Exp. Med. 17:45–53, 2004. © 2004 Wiley-Liss, Inc.
Iodine and selenium are essential components of normal thyroid hormone metabolism and are involved in the modulation of antioxidant defense system. This study was designed to evaluate the extent of peroxidation of lipids and activities of antioxidant enzymes (AOEs) in various tissues of iodine-and/or selenium-deficient rats in relation to thyroid hormone metabolism. Iodine deficiency caused marked enhancements in glutathione peroxidase (GSHPx), superoxide dismutase (SOD), and catalase (CAT) activities in thyroid but did not cause lipid Peroxidation (LP), indicating the Occurrence of an adaptive response that protected the gland against oxidative stress induced by high levels of thyroid stimulating hormone (TSH). Except significant reduction in CAT activity in liver and kidney and an enhancement of SOD in kidney, iodine deficiency did not cause any other alterations in other tissues. Selenium deficiency and combined iodine and selenium deficiency Caused significant alterations in AOE activities in all tissues and caused significantly high levels of LP in thyroid, liver, brain, and plasma. but not in kidney. Alterations in selenium-involved deficiencies appeared to be mainly caused by substantial losses of GSHPx activity; however, compensatory changes in SOD and CAT activities were also observed. (C) 2004 Wiley-Liss. Inc.
Central administration of neuropeptide Y (NPY) produces anxiolytic-like behavioral responses in the conflict test, elevated plus maze, fear-potentiated startle paradigm, and in the chronic immobilization stress. Exogenously administrated NPY also protects against the anxiogenic effects of corticotropin-releasing factor. In the present study, we aimed to determine the effects of centrally administered NPY on the trace element disturbances in brain tissues (frontal and temporal lobes and brain stem) and the other major organs including liver, spleen (zinc [Zn]-, copper [Cu]-, and iron-rich tissues), kidney, and stomach in chronically immobilized rats. The immobilization stress was performed in special cages in which the animals were not able to move. The rats in chronic stress and chronic stress + NPY groups were kept in the cages daily for 7 min for 15 consecutive days. Intracerebroventricular (ICV) cannulas were placed to the right lateral ventricles of the rats by using stereotaxic method. In the control and chronic stress groups, 5 μL of saline (NaCl 0.9%), and in the chronic stress + NPY group, 8 μg NPY/5 μL saline solutions, were administered into the brain via ICV cannula, respectively. Controls and immobilized rats were decapitated 30 min after the injections were over and samples of tissue were taken. Zn, Cu, and iron levels of the frontal lobe, temporal lobe, brain stem, liver, spleen, kidney, and stomach were determined by flame atomic absorption spectrophotometer. Zn and Cu levels were significantly increased in the frontal lobe, temporal lobe, and brain stem in response to chronic immobilization stress daily for 7 min for 15 consecutive days. The administration of NPY inhibited the elevation of Zn in these three parts of brain but did not affect the elevation of Cu in the frontal lobe and brain stem. Increases in Zn and Cu levels of frontal, temporal lobes, and brain stem may be related to induction of MT-I mRNA expression by chronic immobilization stress, and NPY may affect this induction of MT-I, altering corticotropin-releasing factor release in the stress conditions. J. Trace Elem. Exp. Med. 17:283–290, 2004. © 2004 Wiley-Liss, Inc.
In humans, zinc deficiency is characterized by a broad spectrum of neurological clinical syndromes. It is known that vesicular zinc-enriched areas of the brain, such as the hippocampus, are responsive to zinc deprivation, which may result in learning impairment. Recent findings show that zinc deficiency may cause alterations in neurochemical activity. In this study we used contrast-enhanced magnetic resonance imaging (MRI) to monitor disruptions to the blood-brain barrier (BBB) and image-guided MR spectroscopy to follow alterations in brain metabolites as a result of zinc-deficiency and/or hyperoxia-induced oxidative stress. Gadolinium-diethylaminetriaminopentaacetic acid, an extracellular T, relaxation contrast agent, increases tissue water signal in the brain if the BBB is damaged. A significant increase in postcontrast T-1-weighted MR image intensity was observed in the brain of zinc-deficient or hyperoxia-exposed rats, as well as zinc-deficient rats exposed only to hyperoxia when compared with zinc-adequate rats. From single-voxel image-guided MR spectroscopy results, significant decreases in the ratio of N-acetyl aspartate, a neuronal-specific compound, to total choline levels were found When comparing controls (zinc-adequate or zinc pair-fed) with zinc-deficiency or hyperoxia groups alone, and when zinc-deficiency was combined with hyperoxia. This study demonstrates the sensitivity of MR techniques in the ability to monitor the effect of zinc deficiency combined with oxidative stress on BBB permeability as well as detect alterations in brain metabolites. This will further aid in our understanding of the possible cellular and molecular mechanisms involved in zinc deficiency pathology associated with the brain. (C) 2004 Wiley-Liss, Inc.
Melatonin (MEL) is the main neurohormone of the pineal gland. Zinc (Zn) is an essential trace element that is required as a catalytic component for more than 200 enzymes. Both MEL and Zn are considered beneficial for anti-immunosenescence. Recent Findings have shown that MEL can modulate Zn turnover. The aim of this study was to investigate the effect of MEL treatment on the tissue Zn levels in young (4 months) and middle-aged (14 months) rats. Male wistar rats received during 3 weeks subcutaneous injection of MEL (10 mg/kg). After 3 weeks, rats were decapitated and tissue samples were collected. Zn levels were measured by spectrophotometric assay. In conclusion, MEL decreased liver Zn levels both in young and middle-aged rats. In addition, Zn levels in young control group were significantly higher than middle-aged control group. However, MEL treatment increased thymus Zn levels in middle-aged group compared with the control. These findings indicate that tissue Zn levels are significantly affected by MEL treatment.
The purpose of this work was to evaluate insulin sensitivity in chromium mine workers exposed to chromium chronically. Body mass index, waist circumference, fat mass and percent, lipid levels, serum glucose, insulin, leptin and chromium levels, HOMA (%S), and HOMA (%B) values were measured in 93 male workers and 94 age- and body mass index-matched healthy controls. Fat mass and fat percent of workers were higher than control subjects (P < 0;01 and P < 0.0001, respectively). Mean insulin level of the workers was lower than control subjects (6.2 +/- 4.9 muU/mL and 9.38 +/- 5, respectively, P < 0.0001). Mean serum leptin levels was also lower than the control group (8.47 +/- 6.5 ng/mL and 19.0 +/- 10.8, respectively, P < 0.0001). Serum chromium was higher than the control subjects (407.7 +/- 224.2 nmol/L and 4.45 +/- 3.9, respectively, P < 0.00001). Mean HOMA (%S) index of workers was higher than control subjects (114.4 +/- 55.6 and 55.7 +/- 40.6, P < 0.001). Serum high-density lipoprotein cholesterol levels of the workers were significantly higher than the control group (P < 0.02). In conclusion, long-term exposure to chromium may improve insulin sensitivity and beta-cell function. This improvement appears to be a result of low body fat mass and fat percent in body composition. In addition chromium usage may have useful effects in the insulin sensitivity and therefore in development of type 2 diabetes mellitus.
Tannic acid (TA) is naturally occurring polyphenol present in fruits and vegetables. In this study we report that TA inhibits the carcinogenic effect of 7,12-dimethylbenz(a)anthracene (DMBA) in normal and iron-overloaded mice skin. Albino Swiss mice were given iron–dextran and pretreated with a single topical application of tannic acid; after 1 h, tumors were initiated by multiple topically application of DMBA. Appearance, number, and percent tumor incidence were recorded. When compared with the control group, the pretreated groups showed a significantly higher inhibition of tumor incidence. The induction of [3H]thymidine incorporation in cutaneous DNA and lipid peroxidation was inhibited in TA-pertreated animals as compared with the normal control group. Based on this study, we propose that TA significantly inhibits the augmentation potential of iron–dextran. A depletion in lipid peroxidation levels in TA-pretreated groups indicates that excessive generated oxidants in the mice skin tissues may be quenched by TA and because of the chelation of redox active iron and its faster elimination from the body. In conclusion, our data suggest that TA may be an effective chemopreventive agent and may offer protection against iron mediated skin cancer. J. Trace Elem. Exp. Med. 17:21–29, 2004. © 2004 Wiley-Liss, Inc.
The transition metal zinc is an essential element for normal development and function of biological systems including the central nervous system (CNS). Zinc may be an important trace element for the etiology and symptoms of subgroups of autistic children because it is an essential catalytic Or Structural element of many proteins and important for neural activity and the control of neuronal death. The aim of this study was to evaluate zinc status in children with autistic disorder. plasma, erythrocyte, and hair zinc levels were measured by atomic absorption spectrophotometry in the 45 and 39 autistic children, respectively. Forty-one healthy children in the same age group were selected as controls. plasma and erythrocyte zinc levels in the autistic children were found to be significantly lower than normal (P < 0.05). The mean hair zinc level of the patients was also low when compared with that obtained from normal Turkish children. These results may indicate chronic zinc deficiency or metabolic abnormality of zinc in autistic children. Further Studies are required to clarify underlying mechanisms. (C) 2004 Wiley-Liss, Inc.
Green coconut water (GCW; liquid endosperm) is sterile and has been used in the past in remote locations and/or during armed conflicts as a short-term intravenous hydration fluid. It is also conceivable to use GCW for total parenteral nutrition (TPN) under similar circumstances. Patients on TPN need elemental supplementation. Although data on the major elements (calcium, magnesium, potassium, sodium, and phosphorus) found in GCW are abundant, very limited information concerning trace and minor element content in GCW is available. The purpose of this study was to determine trace and minor element content in GCW. Major elements were arbitrarily defined as those with a GCW concentration in the mM range, as opposed to minor ones in the muM range and trace elements in the nM range. The values determined show the great variability typical for samples of plant origin. Under TPN conditions, GCW (1,000 mL/day) would be able to provide adequate supplementation of manganese (12 +/- 5 mumol/L) and possibly zinc (6 +/- 1 mumol/L) but not chromium (not detected, i.e., less than 9 nM/L) and copper (105 +/- 100 nM/L). With respect to elements usually considered nonessential or even toxic, GCW compares favorably with commonly used TPN solutions, most notably for aluminium content (740 +/- 360 nM/L) and cadmium (7 +/- 4 nM/L). The levels of lead (150 +/- 67 nmol/L) and barium (600 +/- 300 nmol/L) were similar to those typically found in commercial TPN solutions. Those of strontium (8 +/- 4 mumol/L) are higher but unlikely to be harmful. In conclusion, although GCW is not completely equivalent to high-tech (and high-cost) TPN solutions with trace element supplements, it still may provide sufficient support of at least some of the trace elements if used as a substitute. The GCW investigated by us is, at least, not toxic. However, our results for several trace elements are much lower than values previously reported in the literature, especially in coconuts from "polluted" areas. GCW from polluted soil might be easily toxic if used intravenously.
Lithium is widely used in medicine as an antidepressive drug. Despite abundant literature on the subject, questions on the effects of lithium on collagen, which is one of the major components of tissues and organs, are far from being answered. We have examined the effects of lithium chloride (1.5 mEq Li/kg of body weight) on rat thoracic descending aorta: 1) on collagen fibrils by electron microscopy and image analysis and 2) on breaking pressure. Animals were sacrificed 1 day, 2, and 12 months after the end of a 30 consecutive days experimental period. In all cases investigated, structural abnormalities in collagen fibrils, at the ultrastructural level, were found. These abnormalities consist of decreased mean diameter and high variability in width as well as marked disorganization in the packing of the fibrils. The observed alterations seem to be permanent. Rupture of the rat thoracic aorta following pressure is significantly influenced by lithium. (C) 2004 Wiley-Liss. Inc.
The present study is designed to investigate the possible role of zinc, copper, manganese, and selenium on the hypoglycemic action of a plants mixture and alfalfa using a diabetic rat model. Diabetes was induced in male Wister rats using streptozotocin and confirmed by estimation of glucose levels in blood and urine and further by histologic examination of the pancreas. The animals were treated with the plants mixture and alfalfa extracts orally by gastric intubation for 7 days. Untreated normal and diabetic rats were included as controls. After evaluating glucose tolerance, animals were sacrificed and organs harvested for trace element analysis and histopathological study. The concentration of zinc, copper, manganese, and selenium in plasma, liver, and pancreas of diabetic rats was significantly lower than those of nondiabetic controls. Treatment of diabetic rats with plants mixture and alfalfa significantly decreased the blood glucose concentration but increased the levels of zinc, copper, manganese, and selenium in plasma, liver, and pancreas. Similar changes occurred in the nondiabetic normal control animals treated with the same products. Histological and immunohistochemical examination also confirmed the presence of more beta-cells in the islets of treated diabetic animals compared with untreated diabetic controls. There was no histological evidence of damage to the liver of either diabetic or normal control animals treated with the plants extract. The present study indicated that treatment of diabetes with naturally available plants mixture appears to be effective and the hypoglycemic effect could be through a multimechanism action including that exerted by the trace element content in the different tissues including the pancreas.
Serum selenium (Se) is a constitutional part of both major plasma selenoprotein P and glutathione peroxidase (GSH-Px), a cytoprotective enzyme against the oxidative damage. It is an accepted fact that any case related to oxidative stress increased by physical exercise changes serum Se levels. In this Study serum Se levels were examined in high physical activity (group I). moderate physical activity (group II), and low physical activity (group III) mates undertaking a soccer-training regimen. In addition, Se changes before and after the acute intensive maximal exercise in an anaerobicloading coordination tests among groups I, II, and III soccer players were investigated. For a minimum of 3 months, the players in group I (n = 12), exercising more than 5 It a week. group II (n = 9), exercising less than 5 h but more than 2 h a week, and group III (n = 11), exercising regularly or irregularly less than 2h were examined. Heart rate monitor 220 (heart rate)-age formula wits used to evaluate the maximal exercising test during the procedure. Serum Se was analyzed by atomic absorption spectrophotometry. The difference between groups before and after the exercise was tested by Wilcoxon test, and the difference varying in the groups was tested by Kruskal-Wallis variance analyse. The relation between heart rate and serum Se was tested by Sperman's rank correlation analyses. After maximal physical exercise, the serum selenium level decreased significantly compared with pre-exercise values (P < 0.05) in group I only. III group 1, maximal and pre-exercise heart rates as opposed to pre- and post-exercise serum Se level were negative correlated (P < 0.05). In conclusion, a nutrition regime rich in selenium may be beneficial for both athletes who exercise regularly and in patients with increasing oxidative stress. (C) 2004 Wiley-Liss. Inc.
Oral treatment of zinc modulates cytokine secretion during the gestation, lactation, and weaning periods. We used a experimental mice model of zinc supplementation during the perinatal stages to Study the effects of this ion over the production of interleukin IL-1alpha,IL-12, and tumor necrosis factor-alpha by peritoneal macrophages. In addition. we determined the gene expression of these cytokines. Zinc (500 mg/L) was orally administered to mice from gestation to lactation (6-week treatment, Zn +) and weaning (9-week treatment). The serum cytokines IL-1alpha and IL-12 were assayed in the offspring at 21 and 42 days after birth. Our results showed a significant (P < 0.01) increase in cytokine production in the Zn + animals at the end of the lactation stage. There was a tendency for the IL-1 concentration to decrease at postweaning nevertheless. IL-12 concentrations were increased in mice at 42 days of age (P < 0.001). The production of IL-1alpha, IL-12, and tunior necrosis factor-alpha in the macrophages supernatants in vitro followed the same tendencies (P < 0.001). Molecular analysis showed an increase of mRNA synthesis in all cases, from 4-fold to 6-fold, in the cytokines analyzed. Our results Suggest that the increase in the proinflammatory cytokines as a result of zinc administration may potentiate Th I cells response, which could lead to the increase of cytokine production in deficient newborns. (C) 2004 Wilev-Liss. hic.
This review of the copper–iron interaction in Wilson's disease was mainly based on ten patients (three females and seven males) studied in our institutes because the genetic tests of ATP7B for Wilson's disease of primary copper toxicosis and HFE for hemochromatosis, the biochemical parameters of copper and iron, and morphological studies on biopsied liver specimens were complete. All patients had hypoceruloplasminemia and hepatic lesions compatible with Wilson's disease. One patient was homozygous and nine patients were compound heterozygous for the mutations in ATP7B, and all patients were free from the major mutation, C282Y, of HFE. The biochemical parameters of iron metabolism were not specific, except for serum ferritin concentration. Judging from the traditional criteria, seven patients had hyperferritinemia. Histochemical iron was stained in the livers of seven patients and histochemical copper was found in nine patients. Microanalysis was more sensitive than histochemistry, detecting copper and iron accumulation in the hepatocellular lipofuscin particles of all patients. Using an improved fixative, intralipofuscin distribution was found to be different between cuprothionein and iron complexes. Iron overload in Wilson's disease might be worsened after treatment because of the close relation to hypoceruloplasminemia, in which the iron efflux from the liver to the circulation is disturbed.
An experiment was performed to test the hypothesis that low dietary silicon affects blood, bone, and urine indices associated with bone formation and breakdown, bone strength and physical characteristics, and the circulating amount of an extracellular matrix protein (osteopontin) involved in bone cell adhesion and activation. A second objective was to ascertain whether ovariectomy (estrogen deficiency) alters the effects of low dietary silicon on bone formation. Female rats weighing about 56 g were assigned to groups of 10 in a factorially arranged experiment. The variables were supplemental dietary silicon at 0 or 35 mg/kg and ovariectomy (estrogen-deficient) or sham operation at the start of the experiment. The basal silicon-low diet contained about 2 mg Si/kg. Low dietary silicon compared with adequate silicon decreased plasma osteopontin concentration, increased plasma sialic acid concentration, and increased urinary helical peptide excretion. Low dietary silicon also affected the response to estrogen deficiency. Ovariectomy increased plasma alkaline phosphatase in the silicon-supplemented, but not in the silicon-low rats. In contrast, ovariectomy decreased liver ornithine aminotransferase in silicon-low but not in silicon-supplemented rats. Ovariectomy increased the urinary excretion of deoxypyridinoline and decreased the femur concentration of sialic acid more markedly in silicon-supplemented than silicon-low rats. Silicon and an interaction between silicon and ovariectomy only mildly changed bone strength and physical measurements and did not affect femur calcium concentration. The findings suggest that silicon has a biochemical function that affects bone growth processes before bone crystal formation by affecting bone collagen turnover and sialic acid-containing extracellular matrix proteins such as osteopontin. J. Trace Elem. Exp. Med. 17:137–149, 2004. Published 2004 Wiley-Liss, Inc.
