In vitro growth inhibition of human cancer cells by novel honokiol analogs

Honokiol possesses many pharmacological activities including anti-cancer properties. Here in, we designed and synthesized honokiol analogs that block major honokiol metabolic pathway which may enhance their effectiveness. We studied their cytotoxicity in human cancer cells and evaluated possible mechanism of cell cycle arrest. Two analogs, namely 2 and 4, showed much higher growth inhibitory activity in A549 human lung cancer cells and significant increase of cell population in the G0-G1 phase. Further elucidation of the inhibition mechanism on cell cycle showed that analogs 2 and 4 inhibit both CDK1 and cyclin B1 protien levels in A549 cells.     

Prognosis prediction of uterine torsion mechanical treatment (rolling) after estimation of calcium and creatinine level in the serum of buffaloes (bubalus bubalis)

The present study was carried out to investigate the relationship between creatinine and calcium concentration in buffalo serum in cases of uterine torsion before rolling, and 1 h and 24 h after calving. The degree, duration and site of uterine torsion, as well as fetus viability, time needed for cervical dilation, and the occurrence of uterine rupture were recorded. A total of 150 pregnant buffaloes suffering from colic and anorexia were brought to our clinic and clinically examined. Of these, 35 buffaloes were diagnosed with uterine torsion (different durations, directions and degrees) and the viability of the foeti and any evidence of uterine rupture were determined using ultrasonography and the serum levels of creatinine and calcium were estimated using calorimetric method. The animals were also examined for incidence of uterine rupture after rolling and calving. The calcium level significantly (P < 0.05) decreased with increasing duration and severity of uterine torsion, however, it was higher in cases where a live fetus was delivered compared with a dead one. Conversely, the creatinine level significantly (P < 0.05) increased with increasing duration and severity of uterine torsion but was lower in cases that delivered a live fetus compared with a dead one. The calcium and creatinine levels returned to approximately normal concentration within 24 h after calving. In conclusion, calcium and creatinine serum concentration have a correlation with duration and severity of uterine torsion. Animals with low levels of calcium (below 8.44 mg/dL) and high levels of creatinine (above 2.25 mg/dL) did not usually respond to rolling or suffer from uterine rupture during calving. The calcium and creatinine levels can be used as indicators for the prognosis of mechanical treatment of uterine torsion in buffaloes.     

Evaluation of Anti-Toxoplasma IgG, IgM, and IgA in Mothers with Spontaneous Abortion in Zanjan, Northwest Iran

Toxoplasma gondii is one of the major agents of infectious abortions and due to its worldwide distribution can threat healthy pregnant women who had no previous exposure to this parasite. The present study was designed to investigate the contribution of T. gondii to spontaneous abortions in Zanjan, Northwest of Iran, using ELISA method. Blood Samples were collected from 264 mothers referred to the provincial hospitals of Zanjan due to spontaneous abortion. The sera were isolated and subjected to evaluate the anti-Toxoplasma IgG, IgM and IgA antibodies. The results showed IgG positive (IgG⁺) in 99 cases (37.5%). A total of 68 women (25.8%) showed seroconversion with IgM or IgA or both IgM and IgA. They included: IgM⁺ in 21 (8.0%), IgA⁺ in 23 (8.7%) and both IgM⁺ and IgA⁺ in 24 (9.1%) subjects. In 23 cases, positive titers of IgM and IgG were accompanied. In general, the analysis of anti-Toxoplasma antibody patterns, showed that about 17% of the spontaneous abortions were associated with serological patterns of acute infection. According to these findings, a considerable proportion of spontaneous abortions can be attributed to T. gondii in the study area.     

Transport of Cryptosporidium parvum Oocysts in Soil Columns following Applications of Raw and Separated Liquid Slurries

The potential for the transport of viable Cryptosporidium parvum oocysts through soil to land drains and groundwater was studied using simulated rainfall and intact soil columns which were applied raw slurry or separated liquid slurry. Following irrigation and weekly samplings over a 4-week period, C. parvum oocysts were detected from all soil columns regardless of slurry type and application method, although recovery rates were low (<1%). Soil columns with injected liquid slurry leached 73 and 90% more oocysts compared to columns with injected and surface-applied raw slurries, respectively. Among leachate samples containing oocysts, 44/72 samples yielded viable oocysts as determined by a dye permeability assay (DAPI [4',6'-diamidino-2-phenylindole]/propidium iodide) with the majority (41%) of viable oocysts found in leachate from soil columns with added liquid slurry. The number of viable oocysts was positively correlated (r = 0.63) with the total number of oocysts found. Destructively sampling of the soil columns showed that type of slurry and irrigation played a role in the vertical distribution of oocysts, with more oocysts recovered from soil columns added liquid slurry irrespective of the irrigation status. Further studies are needed to determine the effectiveness of different slurry separation technologies to remove oocysts and other pathogens, as well as whether the application of separated liquid slurry to agricultural land may represent higher risks for groundwater contamination compared to application of raw slurry.     

Effects of starvation on oxygen consumption, ammonia excretion and biochemical composition of the hepatopancreas on adult males of the False Southern King crab Paralomis granulosa (Crustacea, Decapoda)

Adults of the False Southern King crab, Paralomis granulosa, were starved between 0 and 12 days to evaluate the impact of fasting on the oxygen consumption, nitrogen excretion, O/N ratio and changes on biochemical composition of the hepatopancreas. During the experiment, no mortalities were recorded; physiological changes were detected after 6 days of fasting with an increase of nitrogen excretion (p<0.05). After 9 days of starvation, the crabs showed a maximum decrease in the lipid content (4.3+/-1.2%, p<0.05), accompanied by an increase in oxygen consumption (53.1+/-10.9 microg O2 h(-1) g(-1)). The lowest O/N ratio was detected after 6 days (6.4+/-4.8) and the highest after 12 days of fasting (38.1+/-20.4), indicating that initially crabs utilized proteins as source of energy , followed by lipids. Moreover, after 12 days, there was a significant increase in the hepatosomatic index (HI) and total lipid content (9.7+/-1.0%, p<0.05), which could be associated with the re-absorption of other tissues to the hepatopancreas. Our results provide new information on this species that shows a different pattern of adaptation for each period of starvation and a good correlation between physiological and biochemical parameters. The ability to withstand and recover from periods of nutritional stress is an important adaptation for survival of any organism that must sporadically endure periods of limited food supply.     

Drug resistance in Vibrio cholerae strains isolated from clinical specimens

Cholera is a serious epidemic and endemic disease caused by the Gram-negative bacterium Vibrio cholerae. SXT is an integrative conjugation element (ICE) that was isolated from a V. cholerae; it encodes resistance to the antibiotics chloramphenicol, streptomycin and sulfamethoxazole/trimethoprim. One hundred seven V. cholerae O1 strains were collected from cholera patients in Iran from 2005 to 2007 in order to study the presence of SXT constin and antibiotic resistance.The study examined 107 Vibrio cholerae strains isolated from cholera prevalent in some Iranian provinces. Bacterial isolation and identification were carried out according to standard bacteriological methods. Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) to four antibiotics (chloramphenicol, streptomycin, sulfamethoxazole, and trimethoprim) were determined by broth microdilution method. PCR was employed to evaluate the presence of established antibiotic resistance genes and SXT constin using specific primer sets.The resistance of the clinical isolates to sulfamethoxazole, trimethoprime, chloramphenicol, and streptomycin was 97%, 99%, 99%, and 90%, respectively. The data obtained by PCR assay showed that the genes sulII, dfrA1, floR, strB, and sxt element were present in 95.3%, 95.3%, 81.3%, 95.3%, and 95.3% of the V. cholerae isolates.The Vibrio strains showed the typical multidrug-resistance phenotype of an SXT constin. They were resistant to sulfamethoxazole, trimethoprime, chloramphenicol, and streptomycin. The detected antibiotic resistance genes included dfrA for trimethoprim and floR, strB, sulII and int, respectively, for chloramphenicol, streptomycin, sulfamethoxazole, as well as the SXT element.     

Rational incorporation of selenium into temozolomide elicits superior antitumor activity associated with both apoptotic and autophagic cell death

Background

The DNA alkylating agent temozolomide (TMZ) is widely used in the treatment of human malignancies such as glioma and melanoma. On the basis of previous structure-activity studies, we recently synthesized a new TMZ selenium analog by rationally introducing an N-ethylselenocyanate extension to the amide functionality in TMZ structure.

Principal Findings

This TMZ-Se analog showed a superior cytotoxicity to TMZ in human glioma and melanoma cells and a more potent tumor-inhibiting activity than TMZ in mouse glioma and melanoma xenograft model. TMZ-Se was also effective against a TMZ-resistant glioma cell line. To explore the mechanism underlying the superior antitumor activity of TMZ-Se, we compared the effects of TMZ and TMZ-Se on apoptosis and autophagy. Apoptosis was significantly increased in tumor cells treated with TMZ-Se in comparison to those treated with TMZ. TMZ-Se also triggered greater autophagic response, as compared with TMZ, and suppressing autophagy partly rescued cell death induced by TMZ-Se, indicating that TMZ-Se-triggered autophagy contributed to cell death. Although mRNA level of the key autophagy gene, Beclin 1, was increased, Beclin 1 protein was down-regulated in the cells treated with TMZ-Se. The decrease in Beclin 1 following TMZ-Se treatment were rescued by the calpain inhibitors and the calpain-mediated degradation of Beclin1 had no effect on autophagy but promoted apoptosis in cells treated with TMZ-Se.

Conclusions

Our study indicates that incorporation of Se into TMZ can render greater potency to this chemotherapeutic drug.