LGR6 is a prognostic biomarker for less differentiated tumors in lymph nodes of colon cancer patients.

Introduction: The aim was to investigate whether the stem cell marker LGR6 has prognostic value in colon cancer, alone or in combination with the prognostic biomarkers CEA and CXCL16. Methods: LGR6 mRNA levels were determined in 370 half lymph nodes of 121 colon cancer patients. Ability to predict relapse after curative surgery was estimated by Kaplan-Meier survival model and Cox regression analyses. Results: Patients with high LGR6 levels [LGR6(+)] had a decreased mean survival time of 11 months at 5-year follow-up and 47 months at 12-year follow-up, respectively, with hazard ratios of 3.2 and 2.8. LGR6 mRNA analysis added prognostic value to CEA and CXCL16 mRNA analysis. In the poor prognosis groups CEA(+) and CXCL16(+), further division was achieved by LGR6 analysis. LGR6(+) patients had a very poor prognosis. LGR6 also identified a small number of CEA(-), TNM stage I patients who relapsed suggesting stem cell origin of these tumors. LGR6 and LGR5 levels correlated strongly in lymph nodes of stage I and IV patients but not in stage II patients, suggesting that these stem cell markers are differentially regulated. Conclusion: This study highlights LGR6 as a useful prognostic biomarker independently and in combination with CEA, CXCL16 or LGR5 identifying different risk groups.

Toxic effects of excess exposure to boric acid on serum biochemical aspect, hematology and histological alterations and ameliorative potential role of melatonin in rats.

The current work clarifies the negative effects of excess exposure to boric acid (H3BO3) as a boron-containing compound on rats and the possible ameliorative effect of melatonin (MEL). Forty rats were equally divided into 5 groups as follows: group 1 was treated as control while groups 2, 3, 4 and 5 were orally administered corn oil (0.5 ml), H3BO3 (1330 mg/kg BW), MEL (10 mg/kg BW) and H3BO3 + MEL for 28 consecutive days, respectively. At the end of the experiment, blood was sampled for biochemical and hematological analysis and tissues were collected for histopathological examination. The obtained results demonstrated that the exposure to H3BO3 induced hepatorenal dysfunctions, alterations in bone-related minerals and hormones levels, prostaglandin E2 as inflammatory mediator and hematological indices. H3BO3 induced histological alterations in the liver, kidneys, bone and skin. The co-administration of MEL with H3BO3 resulted in a significant improvement in most of the measured parameters and restoration of morpho-functional state of different organs compared to the H3BO3 group. In conclusion, the study clearly demonstrated that H3BO3- induced various adverse effects and that melatonin may be beneficial in a partial mitigating the H3BO3 and may represent a novel approach in the counteracting its toxicity.

Assessment toxic effects of exposure to 3-indoleacetic acid via hemato-biochemical, hormonal, and histopathological screening in rats.

The current study purposed to investigate the 3-indoleacetic acid (IAA) possible adverse impacts on hematological parameters, hepatorenal function, cardiac, and skeletal muscles as well as testes of rats and histopathological alterations of respective organs and to determine the extent of reversing any adverse impacts occurred in animals after IAA withdrawal. Rats were exposed orally to 500 mg/kg BW by gastric intubation once daily for 14 days, after which one-half was sacrificed and the remaining half left for a further 14 days without IAA exposure. The exposure of rats to IAA produced anemia, leukopenia, neutrophilia, lymphopenia, and a significant increase in activities of serum transaminase, gamma-glutamyl transferase, creatine kinase-myocardial band, creatine kinase-muscle type, and levels of serum creatinine, sodium, chloride, and potassium. Furthermore, serum levels of testosterone, gonadotropins, and leptin significantly declined. The changes in most of measured parameters continued after IAA withdrawal. Histopathological alterations in different tissues supported these changes. In conclusion, subacute exposure to IAA at a high concentration could exert hematotoxicity and toxic effects on many soft organs and its withdrawal led to incomplete recovery of animals. Thus, IAA should be used cautiously as extensive use of it at high concentrations can cause harmful effects on the environment, animals and human beings.

Prognostic Significance of GPR55 mRNA Expression in Colon Cancer.

G protein-coupled receptor 55 (GPR55) probably plays a role in innate immunity and tumor immunosurveillance through its effect on immune cells, such as T cells and NK cells. In this study, the prognostic value of GPR55 in colon cancer (CC) was investigated. mRNA expression levels of GPR55 were determined in 382 regional lymph nodes of 121 CC patients with 12 years observation time after curative surgery. The same clinical material had previously been analyzed for expression levels of CEA, CXCL16, CXCL17, GPR35 V2/3 and LGR5 mRNAs. Clinical cutoffs of 0.1365 copies/18S rRNA unit for GPR55 and 0.1481 for the GPR55/CEA ratio were applied to differentiate between the high- and low-GPR55 expression groups. Kaplan-Meier survival analysis and Cox regression risk analysis were used to determine prognostic value. Improved discrimination between the two groups was achieved by combining GPR55 with CEA, CXCL16 or CXCL17 compared with GPR55 alone. The best result was obtained using the GPR55/CEA ratio, with an increased mean survival time of 14 and 33 months at 5 and 12 years observation time, respectively (p = 0.0003 and p = 0.003) for the high-GPR55/CEA group. The explanation for the observed improvement is most likely that GPR55 is a marker for T cells and B cells in lymph nodes, whereas CEA, CXCL16 and CXCL17, are markers for tumor cells of epithelial origin.

The ameliorative efficacy of Thymus vulgaris essential oil against Escherichia coli O157:H7-induced hematological alterations, hepatorenal dysfunction and immune-inflammatory disturbances in experimentally infected rats.

The purpose of the present study was to evaluate the possible ameliorative role of Thymus vulgaris (T. vulgaris) essential oil against Escherichia coli O157:H7 (E. coli O157:H7) deleterious effects in both blood and different tissues of rats by assessing the hematological, biochemical and immune-inflammatory parameters besides the histopathological alterations in the different organs. Forty male rats were randomly divided into four equal groups as follows: group I served as control, group II orally inoculated with E. coli O157:H7 at a dose of 1.0 × 109 cfu/ml, group III orally received 250 mg/kg BW T. vulgaris oil daily for 7 days and group IV orally inoculated with E. coli O157:H7 as the same dose of group II and orally received T. vulgaris oil as the same dose and duration of group III. Bacterial challenge in groups II and IV was once at the beginning of experiment and administration of oil began after 72 h from bacterial inoculation. At the end of the study, blood was sampled and complete blood picture, liver and kidney function alongside immunoglobulins and cytokines concentrations were estimated and tissues of large intestine (colon), liver and kidneys were collected for histopathological examinations. The results revealed that there was an increase of red blood cells count, hematocrit value and hemoglobin concentration besides white blood cells and thrombocytes counts and substantial increment of serum markers of hepatorenal damage such as the activities of transaminases and concentrations of bilirubin (total, direct and indirect), total proteins, albumin, creatinine and urea in E. coli O157:H7-challenged group. Also, there was a considerable increase in serum immunoglobulins M and G, interleukin 6 and 8 and tumor necrosis factor alpha as well as decreased serum alkaline phosphatase activity. Moreover, T. vulgaris oil could partially improve the hematological, biochemical and histopathological alterations induced by E. coli O157:H7 without any significant alterations in all measured parameters when used alone. The study concluded that the T. vulgaris oil relatively diminished the alterations in hematological parameters, hepatic and renal function markers and immune-inflammatory variables alongside the histopathological changes in different organs induced by E. coli O157:H7. The ameliorative effects of T. vulgaris oil are mediated through its anti-inflammatory, antioxidant and immunomodulatory activities.

The Impact of Iodine Exposure in Excess on Hormonal Aspects and Hemato-Biochemical Profile in Rats.

Excessive exposure of iodine over a time is well known to cause thyroid dysfunction, which may be followed by different effects on body organs. The present study aimed to illustrate the impacts of exposure of rats to excess iodine (above the tolerable range) and the reversibility of any negative impacts on hormonal profile related to thyroid besides cortisol and the hematological and biochemical parameters along with the histopathological alterations in the thyroid gland, liver, kidneys, and heart. Seventy-five rats were divided equally into three groups: Group 1 was control animals. Groups 2 and 3 received sodium iodide (NaI) orally at a dose of (35 and 70 mg/kg BW), which corresponded to (500 and 1000) times excess iodine from the physiological dose, respectively for 30 days, then the NaI administration stopped in the treated groups for 15 consecutive days. Blood and tissue samples were collected twice for various experimental tests after 30 and 15 days of exposure to excess iodine and stopping the exposure, respectively. Overall results revealed that excess iodine in both tested groups developed a hyperthyroid condition, hypercortisolism, relative polycythemia, neutropenia, elevation in serum liver and cardiac enzymes activities, hyperprotenemia, hyperglobulinemia, elevation in serum urea, and cardiac troponin I concentrations (p < 0.05). It was concluded that the excess iodine caused hyperthyroidism, which was associated with significant changes in erythrogram and leukogram and alterations in hepatic, renal, and cardiac functions in an iodine dose-dependent damage relationship and the most of negative impacts continued after stopping the administration.

Clinical Significance of Stem Cell Biomarkers EpCAM, LGR5 and LGR4 mRNA Levels in Lymph Nodes of Colon Cancer Patients.

The significance of cancer stem cells (CSCs) in initiation and progression of colon cancer (CC) has been established. In this study, we investigated the utility of measuring mRNA expression levels of CSC markers EpCAM, LGR5 and LGR4 for predicting survival outcome in surgically treated CC patients. Expression levels were determined in 5 CC cell lines, 66 primary CC tumors and 382 regional lymph nodes of 121 CC patients. Prognostic relevance was determined using Kaplan-Meier survival and Cox regression analyses. CC patients with lymph nodes expressing high levels of EpCAM, LGR5 or LGR4 (higher than a clinical cutoff of 0.07, 0.06 and 2.558 mRNA copies/18S rRNA unit, respectively) had a decreased mean survival time of 32 months for EpCAM and 42 months for both LGR5 and LGR4 at a 12-year follow-up (p = 0.022, p = 0.005 and p = 0.011, respectively). Additional patients at risk for recurrence were detected when LGR5 was combined with the biomarkers CXCL17 or CEA plus CXCL16. In conclusion, the study underscores LGR5 as a particularly useful prognostic biomarker and illustrates the strength of combining biomarkers detecting different subpopulations of cancer cells and/or cells in the tumor microenvironment for predicting recurrence.

Hematobiochemical Disturbances and Oxidative Stress After Subacute Manganese Chloride Exposure and Potential Protective Effects of Ebselen in Rats.

The present study aimed to detect the possible disturbances induced by subacute exposure to manganese chloride (MnCl2) on some biomarkers of hematology, clinical chemistry and oxidative stress, serum iron homeostasis, and ferritin status beside the histopathological alterations in hepatic and renal tissues, and the potential protective effects of ebselen on the Mn toxicity were also evaluated. Forty-eight rats were divided into four groups: Group 1 was used as a control. Groups 2, 3, and 4 were administered of ebselen as a single protective dose (15 mg/kg BW) intraperitoneal, daily manganese chloride (50 mg/kg BW) orally, and ebselen plus manganese chloride, respectively. The administrations were conducted for 30 days. Blood and tissue samples were collected at the end of the treatment for various experimental tests. Results revealed that MnCl2 did not significantly change in erythrogram with leukocytosis and neutrophilia but significantly increased serum aminotransferases and alkaline phosphatase activities, bilirubin (total, direct, and indirect), globulins, triglycerides, total cholesterol, creatinine, urea, manganese, iron and ferritin concentrations and hepatic glutathione, renal malondialdehyde and nitric oxide levels and hepatic superoxide dismutase activity, while serum albumin, hepatic malondialdehyde, and nitric oxide concentrations were significantly decreased besides non-statistical change in serum total proteins concentration. Ebselen has reduced the disturbances in these analytes in combined treatment group. Collectively, subacute exposure to MnCl2 causes disturbance in the leukogram, and hepatic and renal functions with marked renal oxidative stress. It also disturbed serum iron homeostasis and ferritin status. Remarkably, ebselen appears to be highly effective in attenuating the various adverse effects of manganese.

Hematological, biochemical, and histopathological impacts of barium chloride and barium carbonate accumulation in soft tissues of male Sprague-Dawley rats.

The present study was designed to investigate the hematotoxicity, sero-biochemical and histological changes due to the accumulation of BaCl2 and BaCO3, the most important barium salts in our daily lives, in different soft tissues including the liver, kidney, heart, and spleen of adult rats after an oral exposure for 30 consecutive days, and to explain the different mechanisms by which this metal can exert these impacts. For this purpose, adult male rats were divided into three main groups of 15 animals each: group I, serving as controls, group II, receiving BaCl2 orally in a dose of 179 mg barium/kg b.wt, and group III, receiving BaCO3 orally in a dose of 418 mg barium/kg b.wt. for 30 consecutive days. Obviously, normocytic normochromic anemia was evident in both barium groups. Serum biochemical analysis revealed significant declines in glutathione peroxidase, catalase, superoxide dismutase, and urea with significant elevations in malondialdehyde, lactate dehydrogenase, and creatine kinase levels. Hyperphosphatemia, hypokalemia, hypocalcemia, and hypochloremia were also evident in both barium groups. Besides, residual analysis of both barium salts in different body organs revealed significantly abundant barium residues in the liver, spleen, heart, and kidney, respectively in both barium salts groups. Moreover, splenic tissue showed hemosiderosis, peritubular congestion, and necrotic glomeruli with intratubular hemorrhage. Sever subepicardial congestion with intramuscular edema was evident in the heart. In conclusion, BaCl2 and BaCO3 were able to deliver mortalities, antioxidant enzymes exhaustion, and a sort of normocytic normochromic anemia, as well as marked disturbances in cardiac, hepatic, and renal functions due to the accumulation of these two salts in the soft tissues. Therefore, these results demonstrate the unrecognized toxicity of those two barium salts due to their accumulation in various soft tissues of the body and so, this needs to reconsider about barium exposure.

Effect of acute exposure to nonylphenol on biochemical, hormonal, and hematological parameters and muscle tissues residues of Nile tilapia; Oreochromis niloticus.

AIM: This study was aimed to evaluate some biochemical, hormonal, hematological, and histopathological changes in Nile tilapia, Oreochromis niloticus, after acute exposure to nonylphenol (NP). In addition to detection of NP residues in the fish, muscle tissues for human health concern. MATERIALS AND METHODS: A total of 90 apparently healthy Nile tilapia, O. niloticus, were randomly divided into three equal groups; each containing 30 fish (three replicates). Groups 1 and 2 kept as a control and solvent control (acetone), respectively, and Group 3 exposed to NP at a dose level of 500 µg/L water for 7 successive days. Blood and tissue samples were collected 2 times randomly from each group after 7 days from fish exposure to NP and 10 days from exposure stopping. RESULTS: Fish exposed to NP Group 3 showed anorexia, sluggish movement, erythema of the skin, areas of scales loss, and hemorrhagic ulcers in some areas of body region leading to exposing the viscera. Biochemical results revealed a significant increase in serum total proteins and globulins levels, a highly significant increase in serum alanine aminotransferase and aspartate aminotransferase activities, triglycerides, cholesterol, and creatinine levels, insignificant increase in serum uric acid level, and a highly significant decrease in serum testosterone and estradiol-ß17 levels in Group 3 in compare with the control group. Histopathological finding confirms these results. While hematological results of the same group revealed a significant increase in red blood cells count and packed cell volume value, insignificant increase in hemoglobin concentration, leukopenia, lymphopenia, and monocytopenia in compared with the control group. All of these changes appeared after 7 days from fish exposure to NP. Most of these alterations returned toward the normal level after 10 days from stopping exposure to NP. NP residues detected in fish muscle tissues of Group 3 during exposure and after stopping exposure to it. CONCLUSION: It is concluded that NP is a toxic pollutant and has an adverse effect on fish health and reproduction as well as accumulates in fish muscle tissues which may cause human health hazard.

Anti-tumour effects of Egyptian propolis on Ehrlich ascites carcinoma.

A total of 150 female Swiss mice were used to study the ability of water soluble propolis derivatives (WSPD) of Egyptian propolis to inhibit the proliferation and growth of Ehrlich ascites carcinoma (EAC) cells in mice. The mice were divided equally into three groups: the first was kept as a negative control group, the second received an intraperitoneal injection of 2.5 × 10(6) EAC and was kept as a positive control group and the third an intraperitoneal injection of 2.5 × 10(6) EAC and treated with propolis (50 mg/kg body weight) administered by gastric intubations 2 h prior to the intraperitoneal injection of EAC. The propolis was administered daily for 11 successive days. An examination of EAC cells revealed a reduction in the volume, total cell count, viable percentage and increase in the percentage of dead cells in the treated group with an increasing mean survival time (MST), increasing life span (ILS) percentage and treated vs positive control (T/C) percentage. Immunological studies revealed a significant increase in the lymphocyte transformation rate (LTR), phagocytic activity and killing power in the group treated with propolis. A haematological study of the parameters revealed leucocytosis in cancer-bearing mice and propolis-treated groups with granulocytosis and monocytosis. The erythrogram revealed a significant reduction in red blood cell (RBC) count in group 2. The result showed that the implantation of EAC in Swiss mice without treatment resulted in a significant decrease in total protein and albumin levels without a change in globulin level and a significant increase in creatinine level, while the third group that received propolis showed an improvement in these biochemical parameters compared to the normal control group.