Aortic stiffness in normal and hypertensive pregnancy.

The objective of this study was to examine whether aortic stiffness, as assessed by pulse wave analysis, could reliably discriminate between normal and hypertensive pregnancies. One hundred pregnant women were studied: five with severe pre-eclampsia, 27 with gestational hypertension, 14 with chronic hypertension and 54 with normal pregnancy. Central hemodynamic parameters were obtained by an applanation tonometry and included central aortic systolic blood pressure (CSBP), central aortic diastolic blood pressure (CDBP), augmentation pressure (AP), augmentation index (AIx), AIx corrected to a heart rate of 75 (AIx@75) and time to reflection (Tr). All measures of aortic stiffness, including AP, AIx and AIx@75 were significantly higher in women with gestational hypertension and pre-eclampsia compared with normal pregnancies and women with chronic hypertension (p < 0.05 for all comparisons). There were no significant differences between normal pregnancies and women with chronic hypertension (p > 0.05 for all comparisons). Tr was significantly shorter in women with pre-eclampsia and gestational hypertension compared with normal pregnancies (p < 0.05). Aortic stiffness, as assessed by pulse wave analysis, is significantly increased in women with pre-eclampsia and gestational hypertension but not in treated women with chronic hypertension. Pulse wave analysis has a potential as a screening tool in women at high risk for pre-eclampsia. The final role of this method should be determined in prospective studies.

Non- Hodgkin lymphoma of the lips: A rare entity.

AIM: To investigate clinico-pathological features of lymphoma of the lips, and review the literature. MATERIALS AND METHODS: Retrospective analysis and review of English literature, 1996-2016. RESULTS: Analysis included 23 cases, 7 new cases and 16 from literature, 12 M: 11 F, age 7-82 years. Four occurred in children, mean age 10.1; 19 in adults, mean 61.1 years. The lower lip was involved in the majority of cases (16, 69.56%). 14 (60.87%) were isolated to the lips, 8 (34.78%) were multifocal. Nine (39.13%) occurred in association with Sjogren's syndrome, of which one also had Hashimoto thyroiditis. IgG4-related disease and HIV were reported in one case each. The lip salivary glands were involved in most cases (19, 82.6%); 3 (13.6%) showed only cutaneous involvement. The typical presentation was single or multiple nodules (15, 65.21%), with surface ulceration in only two (8.69%). Constituent symptoms were absent in all cases, paresthesia was reported in one (4.34%). The majority (18, 78.26%) was extranodal marginal zone B-cell lymphoma - mucosa-associated lymphoid tissue lymphoma (EMZB-MALT), and one case each was mantle cell, NK-T cell, CD30 positive and plasmablastic lymphoma. CONCLUSION: The lips seem to have a unique pattern of non-Hodgkin lymphoma dominated by EMZB-MALT lymphoma, rarely other types. In more than half, neither Sjogren's syndrome nor other chronic inflammation was identified. Lesions tend to present as asymptomatic slowly progressing, non-ulcerated submucosal masses. Lymphoma should be considered even in the absence of constituent symptoms, as most cases showed none. Although the number of reported cases is rather small, disease course is usually prolonged and prognosis seems to be good.

Quality of life after mandibulectomy: the impact of the resected subsite.

The purpose of this study was to identify the factors that impact the quality of life (QOL) scores of patients undergoing mandibulectomy. All patients with a diagnosis of an oral cavity neoplasm involving the mandible who underwent a mandibulectomy between January 1, 2000 and December 31, 2015 and completed a University of Washington QOL questionnaire (UW-QOL) were included in the study. Fifty-eight patients fulfilled all inclusion criteria and completed the UW-QOL questionnaire. Forty patients (69%) underwent a segmental mandibulectomy and 18 patients underwent a marginal mandibulectomy. Forty-eight patients (82.7%) had a free flap reconstruction. There was no significant difference in the QOL scores between patients who underwent a marginal or a segmental mandibulectomy. In contrast, patients who underwent symphysial resection reported significantly worse scores in various domains compared to patients with body or ramus segmental mandibulectomy. Patients who underwent a segmental mandibulectomy that included the symphysis had worse outcomes in chewing, recreation, health-related and social QOL domains compared to those whose mandibulectomy did not include the symphysis.

Selenite cytotoxicity in drug resistant and nonresistant human ovarian tumor cells.

Our previous studies on selenite cytotoxicity led us to hypothesize that drug resistant tumor cells with high intracellular glutathione will exhibit a high degree of sensitivity to selenite. To examine this we studied the effects of selenite on drug resistant human ovarian tumor (NIH:OVCAR-3) cells in three assays of cytotoxicity: proliferation; cell viability (trypan blue exclusion); and attachment to a solid matrix. The cells were sensitive to low levels of selenite: concentrations as low as 5 microM inhibited cell proliferation and attachment; and viability was decreased by concentrations as low as 20 microM. In each of these assays the NIH:OVCAR-3 cells were more sensitive to selenite than were drug sensitive human ovarian tumor (A2780) cells. These results suggest the potential for the utilization of selenite in the treatment of some drug resistant tumors.

Inhibition of cell attachment by selenite.

Brief pre-exposure of HeLa cells to micromolar concentrations of selenite resulted in a dose-dependent decrease in the rate of their subsequent attachment to a solid matrix (tissue culture dish). Similar low concentrations of selenite also inhibited colony formation, but only when the cells were exposed prior to their attaching to the dish, not when they were exposed after attachment. This indicates that inhibition of cell proliferation by selenite requires exposure to higher concentrations for longer periods of time. In contrast, selenate, selenomethionine, selenocystine, and sulfite did not affect cell attachment, even at significantly higher concentrations. Thus, the inhibition of cell attachment is a specific effect of selenite. Selenite also inhibited the attachment of cells to bacteriological dishes coated with fibronectin, laminin, or collagen, proteins that are components of the extracellular matrix. There was no inhibition when the tissue culture dishes or the protein-coated dishes were pre-exposed to selenite. There was also no inhibition when the cells were exposed to selenite during the attachment process. Thus, pre-exposure of the cells to selenite was necessary for inhibition of attachment. Since cell attachment has been shown to be an important early step in tumor cell invasion and metastasis, these results suggest a novel mechanism of the anticarcinogenic effect of selenite: inhibition of the attachment of tumor cells to the extracellular matrix.

A prevention strategy for circumventing drug resistance in cancer chemotherapy.

The development of drug resistance is considered to be a major cause for the failure of chemotherapy in a number of types of cancer, including ovarian, breast and lung. Most previous research has focused on approaches to reverse drug resistance once it has arisen, that is, on the use of agents which can make drug-resistant tumors more sensitive to chemotherapy. Unfortunately, this approach has thus far met with only limited clinical success. Because of the prevalence of drug resistance in cases of advanced cancer, there exists an urgent need to develop new approaches to dealing with this problem. We have hypothesized the feasibility of an alternative approach: the use of specific agents to prevent the development of resistance before it arises. Our initial studies to examine this hypothesis have focused on ovarian cancer. We have designed both in vitro and in vivo systems in which resistance develops rapidly after exposure of tumor cells or xenografts to melphalan or cisplatin. Using these systems we have shown that two selenium compounds, selenite and selenomethionine are able to prevent the induction of resistance. Furthermore, inclusion of selenite in a chemotherapeutic protocol can result in a significant enhancement of the efficacy of cisplatin in suppressing the growth of human ovarian tumor xenografts. These results have supported the idea that prevention may be a useful new approach to the problem of drug resistance in cancer chemotherapy.

The development of drug resistance by tumor cells in vitro is accompanied by the development of sensitivity to selenite.

The effects of selenite on cell viability and proliferation in a line of drug-sensitive human ovarian tumor (A2780) cells were compared with its effects on a melphalan-resistant derivative of these cells (A2780-ME) which had been developed in vitro (Hamilton et al. (1985) Biochemical Pharmacol., 34, 2583-2586). With the A2780-ME cells there was a 50% decrease in the number of viable cells (i.e. which exclude Trypan Blue dye) after exposure to less than 100 microM selenite for 6 h. In contrast, exposure to more than 300 microM selenite was required to achieve the same effect in the parent line. Similarly, exposure to 10 microM selenite resulted in a 50% decrease in A2780-ME cell proliferation, whereas this treatment had only a small inhibitory effect on proliferation of the parent cells. Thus, the development of melphalan resistance in vitro was accompanied by the development of selenite sensitivity. Pre-exposure of the two cell types to buthionine sulfoximine eliminated the difference in their intracellular glutathione levels, as well as most of their differential sensitivity to selenite. Furthermore, the two cell types did not exhibit a difference in sensitivity to selenodiglutathione, the product of the reaction of selenite with glutathione. Thus, the increase in intracellular glutathione, which has been shown to be responsible for the development of drug resistance in these cells is also responsible for the development of selenite sensitivity.

Sensitivity of melphalan-resistant tumors to selenite in vivo.

Previous studies have demonstrated that melphalan-resistant human ovarian tumor cells exhibit a higher degree of sensitivity to the cytotoxic effects of selenite in vitro than comparable drug-sensitive cells (P.B. Caffrey, G.D. Frenkel, Selenite cytotoxicity in drug resistant and non-resistant human ovarian tumor cells, Cancer Res. 52 (1992) 4812-4816; P.B. Caffrey, G.D. Frenkel, The development of drug resistance by tumor cells in vitro is accompanied by the development of sensitivity to selenite, Cancer Lett. 81 (1994) 59-65). We have now examined the sensitivity of drug-resistant tumors to selenite in vivo. A2780 human ovarian tumor cells, or their melphalan-resistant derivative (A2780ME) cells were injected subcutaneously into nude mice and the resulting tumors were found to be melphalan-sensitive and -resistant, respectively, in vivo. Treatment with selenite (2 mg/kg Se s.c.), which had no overt toxic effect on the animals, resulted in a significant decrease in the rate of growth of the melphalan-resistant tumors, but not on the rate of growth of the drug-sensitive tumors. Thus, melphalan-resistant ovarian tumors are also more sensitive to selenite treatment in vivo.

Effect of selenite on tumor cell invasiveness.

Pre-exposure of HeLa or NIH:OVCAR-3 cells to selenite resulted in a dose-dependent decrease in the ability of the cells to invade a layer of Matrigel, a reconstituted basement membrane preparation. In contrast, selenate, selenomethionine and sulfite had no significant effect on cell invasiveness. Exposure of HeLa cells to selenite also resulted in a decrease in two of the necessary steps of the invasion process, attachment and mobility; in contrast, exposure of OVCAR cells decreased attachment but not mobility. There was an apparent correlation between the processes that are affected by selenite and those that involve the cellular fibronectin receptor (alpha 5 beta 1 integrin).

Inhibition by selenium of DNA and RNA synthesis in normal and malignant human cells in vitro.

Several studies have demonstrated differences between normal and malignant cells in their sensitivity to various effects of selenite. We have compared the effect of selenite on DNA and RNA synthesis in two pairs of normal and malignant human cell lines. One pair of cells, CCL-210 (normal lung fibroblasts) and A549 (lung adenocarcinoma cells), exhibited a large difference in their sensitivity to selenite but no significant difference in their sensitivity to selenodiglutathione. They also had a large difference in the level of intracellular sulfhydryl (SH) compounds. In contrast the other pair of cells, WI-38 (normal fetal lung fibroblasts) and WI-38VA (SV-40 transformed WI-38 cells) both had low levels of intracellular SH compounds and exhibited similar (low) sensitivity to selenite. Our results indicate that differences between normal and malignant cells in their sensitivity to selenite could be due to a difference in the reaction of selenite with intracellular sulfhydryl compounds to form selenotrisulfides.

Rapid development of glutathione-S-transferase-dependent drug resistance in vitro and its prevention by ethacrynic acid.

Exposure of A2780 human ovarian tumor cells to a low concentration of melphalan in vitro for 7 days resulted in the development of melphalan resistance. This resistance was not a stable characteristic of the cells since it was lost after 2 weeks in culture in the absence of drug. The melphalan-resistant cells exhibited significant cross-resistance to cisplatin but only minor cross-resistance to doxorubicin. The resistant cells had elevated levels of glutathione-S-transferase activity and mRNA. Exposure of the cells to the ethacrynic acid resulted in a decrease in enzyme activity as well as a reversal of their drug-resistant phenotype, indicating that the enzyme is involved in the resistance. When ethacrynic acid was present during the 7-day exposure of the cells to melphalan, the development of drug resistance was prevented. This system may serve as a useful preliminary step in screening for agents which can prevent the development of chemotherapy-induced drug resistance in human cancer.

Effects of lead acetate on DNA and RNA synthesis by intact HeLa cells, isolated nuclei and purified polymerases.

The effects of lead acetate on DNA and RNA synthesis have been investigated with intact HeLa cells, isolated nuclei, and purified DNA and RNA polymerases. No inhibition of DNA or RNA synthesis in intact cells was found even after exposure to 0.5 mM lead acetate for 18 hr. In contrast, both DNA and RNA synthesis in isolated nuclei were inhibited by lead (with 50% inhibition at approximately 150 and 80 microM respectively). Similarly, both HeLa DNA polymerase alpha and RNA polymerase II were inhibited, with 50% inhibition obtained at approximately 150 and 20 microM lead acetate respectively. The inhibition of nucleic acid synthesis in isolated nuclei can thus be accounted for by inhibition of the polymerases. The sensitivity of Escherichia coli DNA polymerase I to lead acetate was found to be significantly greater than the HeLa DNA polymerase alpha (50% inhibition at only 10 microM), but the sensitivity of the E. coli RNA polymerase was the same as that of the HeLa enzyme.

Involvement of cellular sulfhydryl compounds in the inhibition of RNA synthesis by selenite.

Selenite has been shown previously to inhibit cellular RNA synthesis. Based upon our previous observation that selenite inhibits purified RNA polymerase only in the presence of a sulfhydryl compound (Frenkel et al., Mol Pharmacol 31: 112-116, 1987), we hypothesized that the inhibition of cellular RNA synthesis by selenite involves endogenous sulfhydryl compounds. We found that depletion of cells of endogenous sulfhydryl compounds, by exposure to diethylmaleate (DEM), virtually eliminated the inhibitory effect of a 1-hr exposure of cells to selenite. This inhibition was restored to normal or higher levels when the selenite was reacted with glutathione or cysteamine prior to addition to the DEM-treated cells. RNA synthesis in DEM-treated cells was inhibited after a 4-hr exposure to higher concentrations of selenite. In contrast to the effect of DEM, specific depletion of the cells of glutathione, by exposure to buthionine sulfoximine, had no effect on the inhibition of RNA synthesis by selenite. These results demonstrate the involvement of endogenous cellular sulfhydryl compounds in the inhibition of RNA synthesis by selenite, but indicate that glutathione, in particular, is not involved in this inhibition.

A comparative study of the effects of mercury compounds on cell viability and nucleic acid synthesis in HeLa cells.

The effects of various mercury compounds on HeLa cell viability and DNA and RNA syntheses in intact cells and in isolated nuclei have been studied. The compounds examined were: methylmercuric chloride, ethylmercuric chloride, dimethylmercury, phenylmercuric acetate, p-hydroxymercuribenzoate, p-hydroxymercuribenzenesulfonate, HgCl2, HgSO4 , Hg(ClO4)2 and Hg2(ClO4)2. All of the compounds except dimethylmercury inhibited colony formation as well as DNA synthesis in intact cells and in isolated nuclei. RNA synthesis in intact cells was inhibited by all the compounds except dimethylmercury, p-hydroxymercuribenzoate and Hg(ClO4)2. In isolated nuclei, alpha-amanitin-resistant RNA synthesis was inhibited by all the compounds except dimethylmercury, alpha-Amanitin-sensitive RNA synthesis was stimulated by some compounds, inhibited by some, and unaffected by others. The effects of two non-mercurial sulfhydryl reagents, N-ethylmaleimide and iodoacetic acid, were also examined. These compounds showed a pattern of effects on nucleic acid synthesis which differed considerably from that of the mercury compounds. Neither compound significantly inhibited alpha-amanitin-resistant RNA synthesis in isolated nuclei, although both inhibited RNA synthesis in intact cells. Iodoacetic acid had no inhibitory effect on DNA synthesis in isolated nuclei but strongly inhibited DNA synthesis in intact cells.

Morbid results of prolonged intubation after coronary artery bypass surgery.

OBJECTIVES: This study evaluated the morbid results of prolonged intubation after coronary artery bypass grafting (CABG). METHODS: Over 30 months, 66 of 1,112 patients undergoing CABG required prolonged intubation. They were matched with 66 patients who did not require prolonged intubation. Preoperative and operative variables were evaluated to determine which would predict prolonged intubation. The postoperative courses were then compared to evaluate the effect of prolonged intubation. The study population was divided into three groups: those who underwent early extubation, but required reintubation (n = 24); those who required initial prolonged intubation, but no reintubation (n = 22); and those who required initial prolonged intubation and reintubation (n = 20). RESULTS: Univariate analysis revealed unstable angina (p = 0.037), elevated creatinine (p = 0.001), reduced FEV(1) (p = 0.019), longer cardiopulmonary bypass time (p = 0.009), and a greater positive fluid balance at 24 h (p = 0.0001) as predictors of postoperative prolonged intubation. Multivariate regression analysis revealed elevated creatinine (p = 0.011), FEV(1) (p = 0.022), and fluid balance (p = 0.001) as predictors of prolonged intubation. The study population had longer ICU and hospital stays (p = 0.0001), with more infectious complications (p = 0.0001) and higher mortality (p = 0. 001). In the subgroups of the study population, patients not requiring reintubation had shorter ICU (p = 0.001) and hospital stays (p = 0.0001), fewer infectious complications (p = 0.0001), and reduced mortality (p = 0.0001). CONCLUSIONS: Patients undergoing CABG with reduced FEV(1), renal failure, and positive fluid balance 24 h postoperatively are at risk for prolonged intubation. Prolonged intubation results in significant acute and midterm morbidity and mortality. Early extubation followed by reintubation further increases morbidity and mortality rates in these patients.

A placebo-controlled comparative evaluation of diclofenac dispersible versus ibuprofen in postoperative pain after third molar surgery.

The analgesic efficacy of single oral doses of drinkable diclofenac dispersible 50 mg was compared with that of ibuprofen 400 mg and placebo in a randomized, double-blind, parallel-group trial in 257 adult patients (245 valid for efficacy) with severe postoperative pain after extraction of an impacted lower third molar. In this study, pain intensity (on a 100-mm visual analog scale) and pain relief from baseline (using a five-point verbal rating scale) were assessed serially during an observation period of 6 hours. Intake of rescue analgesic was permitted in case of insufficient therapeutic effect; however at least 1 hour should have elapsed after test drug consumption. On the main efficacy variable, namely, reduction in the pain intensity score, both diclofenac dispersible (n = 83) and ibuprofen (n = 80) were statistically significantly (P < .01) superior to placebo (n = 82) starting at 20 and 40 minutes, respectively, after drug intake. The active medications were also significantly (P < .01) better than placebo for the secondary efficacy parameters viz. summed pain relief scores over 6 hours (TOTPAR-6); frequency of remedication with a rescue analgesic in the three treatment groups (diclofenac, 24%; ibuprofen, 28%; placebo, 65%); mean time to remedication; and global evaluation. All the treatments were well tolerated. Thus assay sensitivity of this trial (ibuprofen significantly better than placebo) has been demonstrated; in addition, diclofenac as a dispersible formulation has been shown to be an effective analgesic for the treatment of post-surgical dental pain.

Selenium compounds prevent the induction of drug resistance by cisplatin in human ovarian tumor xenografts in vivo.

PURPOSE: The development of drug resistance is a major cause for the failure of chemotherapy, particularly in ovarian cancer. Most previous research has focused on approaches to reverse drug resistance once it has arisen, that is, on the use of agents which can make drug-resistant tumors more sensitive to chemotherapy. We have suggested the feasibility of an alternative approach: the use of specific agents to prevent the development of resistance. METHODS: We designed an in vivo system to assay for the ability of compounds to prevent the induction of resistance by cisplatin. In this system, mice bearing tumors (which originated from A2780 human ovarian tumor cells) were treated with a low dose (2.6 mg/kg) of cisplatin and the tumors rapidly developed resistance to subsequent cisplatin treatment. Cell lines initiated from these tumors retained the resistant phenotype even after several months in culture. RESULTS: When either selenite or selenomethionine were administered (i.p., 1.5 mg/kg) close to the time of the initial cisplatin treatment, the induction of resistance was prevented. Similar treatments with sulfite or methionine had no effect on the induction of resistance by cisplatin. Studies in cells from treated tumors have indicated that the selenium compounds may prevent the induction of resistance by preventing a cisplatin-induced increase in glutathione level. CONCLUSIONS: Selenium compounds specifically prevent the induction by cisplatin of drug resistance in human ovarian tumors in vivo.

Oxidative and glycolytic metabolism of semen components by washed guinea pig spermatozoa.

The concentration of several potentially metabolizable substances in guinea pig semen and the ability of these substances to support ATP synthesis and the motility of guinea pig sperm have been determined. Both glucose and fructose were present in high concentration in semen and were equipotent at the concentration tested in maintaining high levels of ATP and a high rate of motility. Lactic and pyruvic acids also supported a high rate of sperm motility but maintained lower levels of ATP. These constituents of guinea pig semen, as well as the metabolites alpha-glycerophosphate, succinic acid, and beta-hydroxybutyric acid, are oxidized at unusually high rates. The active oxidative metabolism of guinea pig sperm is compared with that of human sperm which is primarily glycolytic.

Treatment of human ovarian tumor xenografts with selenite prevents the melphalan-induced development of drug resistance.

Human ovarian tumors (derived from A2780 cells), growing as subcutaneous xenografts in immunodeficient mice, develop melphalan-resistant cells after a single treatment of the tumor-bearing animal with the drug [Caffrey, Zhang and Frenkel, submitted for publication]. Treatment of the animals with selenite by i.p. injection prevented the development of primary resistance to melphalan as well as cross-resistance to cisplatin. Selenite treatment also prevented the melphalan-induced increase in the cellular level of glutathione. In contrast, selenite administered s.c. or in drinking water had relatively little effect on the development of resistance. The results in this animal model suggest that selenite may be clinically useful in preventing the development of drug resistance during chemotherapy of cancer.

Rapid development of drug resistance in human ovarian tumor xenografts after a single treatment with melphalan in Vivo.

Human ovarian tumors from A2780 cells were grown as xenografts in immunodeficient mice, treated with a single i.p. dose of melphalan and tumor cells were removed and placed into tissue culture. The cells from the treated tumors exhibited an approximately 2-fold resistance to melphalan in vitro compared to cells taken from untreated tumors. This degree of resistance was similar to that of cells from tumors formed from melphalan-resistant A2780-ME cells. The cells from the treated tumors were also resistant to cisplatin but not to doxorubicin. They contained approximately 2-fold higher levels of glutathione than cells from the untreated tumors. Exposure of the cells to buthionine sulfoximine (a specific inhibitor of glutathione biosynthesis) eliminated the difference in glutathione levels as well as the difference in sensitivity to melphalan. When tumor-bearing animals were treated with buthionine sulfoximine in addition to melphalan the resulting tumor cells were not resistant to the drug. Resistance could also be demonstrated in the tumors themselves in vivo: the growth of previously untreated tumors was severely inhibited by a high dose of melphalan (11.7 mg/kg) administered i.p. to the animals, whereas the growth of tumors which had received prior treatment with melphalan was unaffected by the subsequent high dose. The rapid development of drug-resistant tumor cells after a single drug treatment in vivo makes this an excellent system for the investigation of the mechanisms by which resistance develops as well as for use in the screening for agents which can prevent it.