[Exchange mechanism of chromosome loss].

Frequencies of induced losses of autosome 2, free arm of autosome 2 and simultaneous losses of two free arms of autosome 2 were determined as 3.25 x 10(-3), 2.78 x 10(-3), 0.84 x 10(-3), respectively. High frequency of simultaneous losses of two chromosomes may have pointed to the exchange nature of the chromosome losses. The similarity of frequencies of half-translocations in heterochromatin of autosome 2 and frequencies of the losses observed show that the chromosome losses are the consequence of exchanges in chromosomal heterochromatin.


[Electron microscopic analysis of synaptonemal complexes of male laboratory mice exposed during the period of embryogenesis in the vicinity of the Chernobyl Nuclear Power Station].

Four male mice were shown to have decreased fertility among 74 laboratory male mice exposed in the Chernobyl APP area during embryogenesis. Electron microscopic analysis of synaptonemal complexes (SC) of at zygotene-diplotene and light-optic analysis of chromosome aberrations at diakinesis-metaphase 1 demonstrated the presence of interchromosome translocations in autosomal chromosomes, and also intrachromosome translocations in two of four animals. The frequency of chromosome translocations in SC preparations was on the average 1.8 times higher than their frequency at the diakinesis-metaphase 1 stage. High percentage of cells in which associations of sex bivalent axes (XY) with the axes of autosomal bivalents were observed in the SC preparations and reduced fertility in the animals studied confirmed the Forejt's hypothesis that such associations caused the arrest of cells at pachytene and, as a consequence, disturbed the process of embryogenesis.


[DNA repair in cells of the radiosensitive mutant of Drosophila rad(2)201GI after gamma-irradiation].

A radiosensitive mutant of Drosophila melanogaster rad(2)201GI was analysed for the capacity to repair DNA single- and double-strand breaks induced by gamma-rays. Analysis was performed on cell cultures derived from embryos of homozygous mutant stock and wild type strain Oregon R. The viability of irradiated cells was studied. It was shown that the mutant strain cells had increased lethality, just like a whole organism. Single-strand breaks were analysed by alkaline sucrose gradient centrifugation; double-strand breaks were monitored by neutral elution. The similarity of repair kinetics of single- and double-strand breaks in cells of rad(2)201GI and Oregon R was shown. Probable molecular mechanisms of rad(2)201GI mutant radiosensitivity are under discussion.


[Effect of mutations in the genes su(Hw) and mod(mdg4) on transvection between alleles of the Yellow locus in Drosophila melanogaster].

A typical example of transvection is a complementation between alleles in the yellow locus: y2 (mdg4 insertion inactivating certain y-enhancers) and y1 (deletion of the y-promoter but not of the enhancer). Transvection was explained by trans-activation of promoter in y2-allele by enhancer of y1-allele. Here we found that the mutation mod(mdg4)1u1 in the modifier of mdg4 locus (a regulatory gene controlling, together with suppressor of Hairy wing) expression of (mdg4) completely suppress the complementation. Removal of an acidic domain from su(Hw) protein product in su(Hw)j mutation partially suppress the complementation. We also have found that mod(mdg4)1u1 mutation trans-inactivates the yellow allele with a wild type phenotype (y+2MC) in heterozygote with the y2 allele, i.e. the negative transvection takes place. In this case, deletion removing an acidic domain even in one copy of su(Hw) suppresses the effect of mod(mdg4)1u1 mutation.


[Selection in the context of provocation in artificial populations of Drosophila melanogaster].

The experimental potentials of the method for selection against provocative background (autoselection) in artificial populations of Drosophila melanogaster were analysed. The dynamics of adaptation of Drosophila population under increasing NaCl concentration in food was studied. It appeared that selection was successful in case of feedback between the selection pressure, mediated by alteration of salt concentration, and the rate of flies adaptation determined by the intensity of their reproduction. Within a short period of time, as the result of autoselection, the flies' ability for development with NaCl of up to 8% concentration in food was registered. It was shown that this resistance of flies to the salt was inherited within twelve successive generations.


Characterization of a major tandemly repeated DNA sequence (RBMII) prevalent among many species of waterfowl (Anatidae).

We have investigated the evolution of a 190 base pair tandemly repeated DNA sequence (RBMII) in 27 different species of waterfowl. In this paper we show that the RBMII sequence is present in many species belonging to 7 of the 11 Anatid tribes. Inter- and intra-tribal differences in repeat presence indicate that, although the RBMII sequence has been maintained among widely divergent species, it is rapidly evolving. Restriction enzyme analyses suggest very different hierarchical repeat organizations among different species. DNA sequence comparisons of 32 cloned monomer units from five different species revealed what appears to be a nonrandom distribution of sequence divergence, as well as large differences (up to 25-fold) in intraspecific sequence variation between relatively closely related species.


The phylogeny of nine species of the Drosophila obscura group inferred by the banding homologies of chromosomal regions. III. Element D.

The phylogenetic relationships among nine species belonging to the obscura group of the genus Drosophila were deduced, based on similarities of the banding pattern of their polytene chromosomal element D. These similarities were inferred by the comparison of chromosomal photomaps. The phylogenetic reconstruction was the most parsimonious based on seriation by overlapping inversions and on the principle of conservation/disassociation of nearby located segments. The gene sequences of element D for all species studied were relatively easy to recognize in terms of the map of D. obscura, already found to occupy a relative central position in this group. Thus, three clusters of closely related species could be identified: obscura (D. obscura, D. ambigua, and D. tristis), African (D. kitumensis and D. microlabis), and subobscura (D. subobscura, D. madeirensis and D. guanche), with D. subsilvestris standing apart. The results are in agreement with those from the previously studied elements B and E, but element D was found to be much more conclusive concerning the links among the different clusters. Thus, it is inferred that D. guanche occupies an intermediate position between the other two species of its own cluster and all the others. The gene arrangement of D. obscura, directly related to those of the other species, has been identified. In the phylogenetic tree proposed, both the African cluster and D. subsilvestris derive from a hypothetical gene arrangement, intermediate in the pathway between the subobscura and obscura clusters.


The mitochondrial NADH dehydrogenase subunit 6 (ND6) gene in Murres: relevance to phylogenetic and population studies among birds.

The nucleotide sequences of the mitochondrial ND6 and tRNA(Glu) genes and part of the displacement loop region in two closely related seabird species are presented. A chicken type gene organization in which the tRNA(Glu), ND6, and displacement loop are localized next to each other was found in these species and suggests that this is a conserved feature of avian mitochondrial DNA. The nucleotide and amino acid divergences of ND6 at different taxonomic levels are assessed, and its relevance to phylogenetic studies in birds is discussed.


Silver staining two types of meiotic nodules.

We have developed a reliable method for silver staining nodules on synaptonemal complexes (SCs) of tomato (Lycopersicon esculentum). This technique involves hypotonically bursting primary microsporocytes, fixing SC spreads with paraformaldehyde, and incubating the spreads at 40 degrees C in a 33% aqueous silver nitrate solution covered with nylon mesh. When tomato SCs were stained by this method, nodules were observed with the same distribution and frequency as nodules stained with uranyl acetate and lead citrate. Incubation in silver nitrate at higher temperatures caused the loss of some or all nodules. The pattern of loss suggests that two types of nodules coexist during late zygonema and early pachynema and that one type becomes the late nodules of mid-pachynema through early diplonema.


A distribution model for heritability.

A regression model to predict quantiles of narrow sense individual and family mean heritabilities is developed and used to predict confidence intervals either directly or via a generalized beta distribution model. Extensive simulations of balanced sib analysis trials in randomized complete block designs and normal distributed environmental and additive genetic effects confirmed that heritabilities follow a beta distribution even in cases with up to 10% of the data missing at random. The new model is both more accurate and more precise than commonly used alternatives based on "exact" chi 2 distributions and Satterthwaites approximations to the degrees of freedom. Estimates of the expected heritability and a Taylor approximation of the standard error of the heritability are needed as input to the quantile model. Applications of the presented models for estimating confidence intervals and as an aid in the design of experiments are provided.


Mitochondrial and ribosomal DNA variation among members of the Anopheles quadrimaculatus (Diptera: Culicidae) species complex.

The extent of intra- and inter-specific variation in mitochondrial DNA and nuclear ribosomal RNA gene restriction sites was determined for the four sibling species of the Anopheles quadrimaculatus complex. Individual mosquitoes were identified by allozyme analysis according to previously published keys, and the total genomic DNA of these same individuals was then cleaved with restriction enzymes. Restriction maps of mitochondrial DNA, including the positions of variable sites, were constructed for each species. No evidence for interspecific hybridization was found in the populations surveyed. There was little variation in restriction patterns within any given species, but differences occurred among the four. Three restriction enzymes (AvaI, HindIII, and PvuII) yielded species-specific DNA restriction patterns for the mitochondrial DNA, while AvaI and HindIII produced diagnostic patterns for the ribosomal DNA. Thus, restriction patterns were very useful for detecting cryptic species but less appropriate than isozymes for studying genetic structure of populations within species.


The evolutionary history of Drosophila buzzatii. XXIII. High content of nonsatellite repetitive DNA in D. buzzatii and in its sibling D. koepferae.

The frequency and types of repetitive nonsatellite DNA of two sibling species of the repleta group of Drosophila, D. buzzatii, and D. koepferae have been determined. For each species, the analysis is based on a sample of more than 100 clones (400 kb) obtained from genomic DNA. A theoretical model has been developed to correct for the presence of a mixture of repetitive and unique DNA in these clones. After correction, a high content of repetitive DNA has been demonstrated for both species (D. buzzatii, 19-26%; D. koepferae, 27-32%). The repetitive sequences have been classified according to their hybridization pattern when used as probes against genomic DNA and by their in situ hybridization signals on polytene chromosomes. Data suggest that the main nonsatellite component of these species is simpler and more repetitive than that of D. melanogaster, pointing to a wide variability in content and class size distribution of repetitive DNA among Drosophila species.


Immune reactivity in aging-autologous mixed lymphocyte responses.

The level of activity in the autologous mixed lymphocyte reaction (AMLR) was studied in populations of young and old subjects. AMLR activity was reduced in the older age group. A subpopulation of the older age group who had Alzheimer's disease was shown to have the lowest AMLR values. Within this group, those with a history indicating a more rapid development of CNS-related disability from senescence showed the weakest AMLR responses. The AMLR values correlated neither with sex, nutritional status nor history of infections. This impairment of a central regulating immune reaction may be a significant variable in manifestations of the aging process.


Age-associated decrease in vasopressin-induced renal water transport: a role for adenylate cyclase and G protein malfunction.

Aging is associated with a decline in renal concentrating ability in response to dehydration-induced arginine vasopressin (AVP) release. To examine target tissue sensitivity, cortical collecting tubules (CCT) were individually microdissected from young (3 months), middle-aged (2-3 years) and old (4-5.5 years) rabbits and subjected to in vitro perfusion analysis and tissue culture of defined epithelial monolayers. Osmotic fluid transport was compared with adenylate cyclase activity and an age-associated decline in both parameters detected in response to increasing doses of AVP. [3H]-AVP binding was unchanged in CCT epithelia of different ages, excluding receptor alterations as the underlying mechanism. Also, hydraulic conductivity measurements of isolated young and old CCT were not significantly different, excluding post-cAMP events as a major mechanism of reduced sensitivity. Effects of cholera toxin and forskolin on hydraulic conductivity and adenylate cyclase activity showed a dramatic decrease in the ability of these compounds to elicit a response in CCT epithelia from old rabbits. It was concluded that alterations in Gs proteins and the catalytic unit of adenylate cyclase-were responsible for the age-associated decline in CCT response to AVP.


Event-related potentials in patients with multiple lacunar infarcts.

We studied event-related potentials (ERPs) occurring in response to attended and unattended stimuli in 31 patients with multiple lacunar infarcts. ERPs were recorded during the performance of visual discrimination tasks using three kinds of stimuli. Each component of P300 response to infrequent nontarget stimuli and P300 response to infrequent target stimuli was defined as non-target P3 or target P3. The non-target P3 latency in patients with multiple lacunar infarcts was significantly longer than in 15 age-equivalent normal subjects, while no significant differences could be found in target P3 latency between patients and normal subjects. From the controlled/automatic standpoint these results suggest that multiple lacunar infarcts are related to impairment of automatic processing reflected by non-target P3, although controlled processing reflected by target P3, is less impaired.


Prevalence of peripheral arterial disease and related risk factors in elderly institutionalized subjects.

Blood pressure was measured at the posterior tibial artery by Doppler ultrasonography in 124 elderly subjects (37 men and 87 women) living in two retirement homes in Naples (Italy). Ankle-to-arm systolic pressure ratios below 0.97 and 0.90 was considered as a probable and definite pathological sign of peripheral arterial disease, respectively. Half of the subjects (48% of men and 51% of women) gave a value below 0.97, while a third (35% of men and 33% of women) had a value below 0.90. By multiple regression analysis, mean blood pressure, plasma cholesterol levels and cigarette smoking were all negatively correlated with ankle systolic pressure values with a progressively lower significance. Most of the patients with pathological Doppler examination were asymptomatic at a questionnaire for intermittent claudication.


Physiological factors associated with injurious falls in older people living in the community.

Performance in six tests of sensorimotor function was measured in 50 subjects who were admitted to an acute hospital because of a fall (ICD codes E880-888), but who did not suffer a fracture of the lower limbs as a result. Performances in these tests were compared with 50 subjects of the same age and sex who had not fallen in the previous 12 months. It was found that those admitted to hospital because of a fall had decreased tactile sensation, reduced quadriceps strength and increased body sway on firm and compliant surfaces. The fallers also performed poorly in clinical tests of static and dynamic balance. Psychoactive-drug use was associated with falling and a number of test measures, including body sway, static balance, dynamic balance and quadriceps strength. Twenty-seven percent of fallers had poor outcomes, in that 1 year after testing, they had either suffered three or more additional falls, been readmitted to hospital, been transferred to nursing homes or died.


Nuclear factor kappa B: an oxidative stress-responsive transcription factor of eukaryotic cells (a review).

NF-kappa B is a multiprotein complex that can activate a great variety of genes involved in early defence reactions of higher organisms. In nonstimulated cells, NF-kappa B resides in the cytoplasm in an inactive complex with the inhibitor I kappa B. Pathogenic stimuli cause release of I kappa B and allow NF-kappa B to enter the nucleus, bind to DNA control elements and, thereby, induce the synthesis of mRNA. A puzzling feature of NF-kappa B is that its activation is triggered by a great variety of agents. These include the cytokines interleukin-1 and tumor necrosis factor, viruses, double-stranded RNA, endotoxins, phorbol esters, UV light and ionizing radiation. We recently found that also low concentrations of H2O2 activate NF-kappa B and that various antioxidants prevent the induction by H2O2. Subsequent analysis revealed that antioxidants not only suppress the activation of NF-kappa B by H2O2 but by all other inducers tested so far. In this review, we will discuss the evidences that NF-kappa B is an oxidative stress-responsive transcription factor of higher eukaryotic cells.


Antioxidant vitamin supplementation of smoke-exposed rats partially protects against myocardial ischaemic/reperfusion injury.

Our previous studies showed that exposure of rats to limited periods of cigarette smoke resulted in more severe myocardial damage when their hearts were subjected to myocardial ischaemia/reperfusion. The aim of this study was to determine whether supplementation of rats with antioxidant vitamins alpha-tocopherol and beta-carotene was able to protect their hearts against the increase in ischaemia/reperfusion injury caused by smoke-exposure. The parameters measured were mitochondrial oxidative function, cellular levels of alpha-tocopherol and low molecular weight iron (LMWI). Supplementation with antioxidant vitamins resulted in significantly less mitochondrial functional oxidative damage compared to that observed in the controls. Supplementation did not affect the cellular LMWI content, suggesting that the generation rate of hydroxyl radicals was similar in both groups. The protective effect of alpha-tocopherol and beta-carotene supplementation on the mitochondrial function against ischaemia/reperfusion could be due to their free radical scavenging action. Supplementation with antioxidant vitamins, therefore, had a beneficial effect on the excessive myocardial ischaemia/reperfusion injury of smoke exposed rats.


Glomerular injury induced by hydrogen peroxide: modifying influence of ACE inhibitors.

The sensitivity of isolated glomeruli from normotensive (Wistar-Kyoto, WKY) and spontaneously hypertensive (SHR) strains to oxidant stress was studied by determining the incidence of pyknosis, karyohexis and karyolysis after incubation with different concentrations of hydrogen peroxide (H2O2) (4.7 x 10(-9) - 10(-3) M). Even though the proportion of glomeruli containing nuclei that demonstrated these features increased progressively with increasing concentrations of H2O2, the number of severely damaged glomeruli was relatively small even at concentrations of 4.7 x 10(-3) M. Examination of the surface epithelial cells of glomeruli using scanning electron microscopy revealed no evidence of disturbance of the macroscopic or podocyte structure or, of increased blebbing after H2O2-treatment. These data suggest damage to nuclei is an early result of ROS stress on glomeruli. Preincubation of WKY glomeruli with captopril or lisinopril resulted in a significant drop in the proportion of WKY glomeruli demonstrating structural damage after oxidant stress. In contrast, preincubation of SHR glomeruli with lisinopril had no effect on oxidant-induced changes in the morphology of SHR glomeruli, whereas captopril effected a significant increase in the proportion of glomeruli demonstrating damage at all concentration of H2O2.


Hexamethylmelamine as second-line therapy in platin-resistant ovarian cancer.

A total of 61 patients with recurrent or persistent clinically measurable platin-resistant epithelial ovarian carcinoma were treated with 260 mg/m2 oral hexamethylmelamine daily for 14 days, repeated at 4-week intervals. Platin resistance was defined as progression or stable disease during cis- or carboplatin treatment (used alone or in combination with other drugs), or relapse within 6 months after the end of that therapy. Fifty patients were evaluable for response and 57 for toxicity. The objective response rate was 14% (3 complete and 4 partial responses). The response rate was higher in patients with relapse within 6 months than in patients with progression or stable disease on platin-based therapy. This observation underscores the importance of defining response and time to progression after first-line chemotherapy. The median duration of response was 8 months and the median survival in responding patients was 9+ months versus 5 months for patients with progression on hexamethylmelamine. Nausea and vomiting requiring antiemetic treatment occurred in 8 (14%) patients and reversible peripheral neuropathy in 3 patients. Two patients developed agitation, insomnia, and depression during hexamethylmelamine therapy. In conclusion, the 14% objective response rate and the occurrence of complete responses with oral hexamethylmelamine treatment in a group of ovarian cancer patients with true platin resistance are noteworthy.


Diagnostic value of conization of the uterine cervix in the management of cervical neoplasia: a review of 756 consecutive patients.

A review of 584 consecutive cases of conizations performed over a 15-year period showed that 320 patients (54.8%) had conizations because of the presence of neoplastic cells in the endocervical curettings. Of these patients, 268 (83.4%) had cervical intraepithelial neoplasia of various grades on conization, and 12 (3.6%) had microinvasive cancer. The remaining 40 patients had normal cervical epithelium. One hundred seven (18.3%) patients had conization because of cytological smear and colposcopy discrepancy. Seventy-six (71.0%) patients in this category had cervical intraepithelial neoplasia. Two (2.0%) had superficially invasive cancer; the remaining 29 patients had normal cervical histology. Sixty-seven (11.5%) had conizations because of unsatisfactory colposcopic evaluations. This review indicates that although conization is less frequently performed, it is still a useful tool in the management of patients with abnormal cytology in whom colposcopy evaluation cannot rule out invasive cancer.


Long-term survival with advanced ovarian cancer: an analysis of 5-year survivors in the Australian trial comparing combination versus sequential chlorambucil and cisplatin therapy.

Of 369 patients with apparent advanced ovarian cancer entered on a randomized trial between September 1979 and June 1983, 56 survived more than 5 years. Central pathology review confirmed the diagnosis in 318 cases (15 cases not ovarian primary, 36 slides did not reach central review). Two hundred ninety-eight were invasive cancer and 34 of these (11.4%) patients were alive at 5 years. The remaining 20 cases were tumors of low malignant potential and 17 (85%) were alive at 5 years. The clinical, pathological, and treatment characteristics of these long-term survivors has been ascertained. No significant difference in survival has emerged between the two treatment arms being compared--combination chlorambucil and cisplatin versus sequential chlorambucil followed by cisplatin on treatment failure. Of 298 cases of invasive cancer there were 133 with residuum less than 2 cm after initial surgery and 20 of these (15%) survived 5 years compared with 14/165 (8.5%) of those with more tumor residuum. Clinical response was a poor indicator of survival. Only 7 of 46 (15%) patients with complete clinical response survived 5 years; however, of 44 patients with complete surgical response, 29 (65.9%) were alive at 5 years. These results highlight the improved survival prospects of women with low-potential malignancy, even in advanced stage, compared to those with invasive tumors and make a strong case for central pathology review of all cases of apparent ovarian cancer entered on randomized trials.


Uterine papillary serous carcinoma: a study on 108 cases with emphasis on the prognostic significance of associated endometrioid carcinoma, absence of invasion, and concomitant ovarian carcinoma.

One hundred eight cases of uterine papillary serous carcinoma (UPSC) were analyzed to characterize its histologic features and biologic behavior. Special situations that could conceivably modify the behavior and therapeutic approaches were considered: (1) the occurrence of areas of endometrioid carcinoma in otherwise typical UPSC; (2) the confinement of UPSC to an otherwise benign endometrial polyp or the endometrial mucosa or absence of residual tumor at the time of hysterectomy; and (3) the coexistence of a superficial UPSC and a serous ovarian carcinoma. There was coexistence of endometrioid and UPSC in 22 cases, and tumor was confined to an endometrial polyp or endometrium in 19 cases. There was simultaneous pathologic stage I UPSC and papillary serous ovarian carcinoma in 10 cases. In patients with pathologic stages I and II UPSC the presence of areas of endometrioid carcinoma intermixed with the UPSC did not improve survival. Patients with stage I disease and no residual tumor or tumor confined to an endometrial polyp/endometrial mucosa and without vascular invasion had a survival not statistically different from those with stage I disease but with myometrial and/or vascular invasion. Patients with stage I UPSC with concomitant ovarian serous surface papillary carcinoma had survival not statistically different from patients with stage IV UPSC.


Estrogen and progesterone receptor assay in carcinoma of the cervix with monoclonal antibodies.

The normal cervix has been shown to contain estrogen and progesterone receptors (ER and PR), but there have been controversial reports on the presence of these receptors in cervical carcinoma. Thus to explore the relationships between the steroid receptor status and cervical carcinoma, tissues of 34 patients with invasive cervical carcinoma prior to treatment and 30 cases of control who underwent hysterectomy due to benign gynecologic diseases at Yonsei Medical Center were analyzed for ER and PR using immunocytochemical assay with monoclonal antibodies and the results were compared with those of conventional steroid binding assay. ER and PR were positive in 65 and 71% of all tumors, respectively, and 59% of the cases were positive for both receptors simultaneously. No significant difference in receptor levels was noted when stratified according to menopausal status or clinical stage. With regard to tumor size, lesions greater than 3 cm had significantly lower receptor positivity when compared with the control. For the histological cell types, a statistically significant higher mean ER level was noted in adenocarcinoma compared to squamous cells. Even though the potential therapeutic significance of these findings is not yet known, they are consistent with the concept that steroid receptors can be used as a guide to endocrine therapy with respect to other prognostic parameters. However, the small number of advanced-stage carcinomas and the absence of data regarding survival rate in this study preclude any definite conclusions.


Molar villous fluid suppresses mononuclear cell cytotoxicity.

Complete molar pregnancy tissue is an allograft to the mother because all molar chromosomes are of paternal origin. Interactions between molar tissue and the maternal immune system may be important in the natural history of complete molar pregnancy. Molar villous fluid (MVF) has previously been demonstrated to suppress both mitogen and interleukin-2-induced T lymphocyte proliferation. The current study was undertaken to evaluate the potential effect of MVF on the cytotoxic activity of mononuclear cells (MNC) and lymphokine-activated mononuclear cells (LA-MNC). Sera and molar villous fluid were obtained from four women at the time of molar evacuation. K-562 erythroblastoid cells were used as target cells for MNC-mediated lysis, and JEG-3 choriocarcinoma cells were used as targets for LA-MNC-mediated lysis in a 51Cr release assay. Relative to patient sera, all MVF tested significantly inhibited both MNC and LA-MNC lysis of target cells (48.3 and 91% mean inhibition, respectively; P < 0.05). This study provides additional evidence that molar gestational tissue produces factor(s) that suppress maternal immunologic responses. Potential therapies may become available to reduce or eliminate the immunosuppressive effects of molar gestations resulting in a more favorable clinical outcome in patients with complete molar pregnancy and postmolar gestational trophoblastic tumors.


Steroid receptors in ovarian carcinoma: immunohistochemical determination may lead to new aspects.

87 nonpretreated stage III/IV ovarian common epithelial carcinomas were studied for estrogen receptor (ER) and progesterone receptor (PR) content by both immunohistochemistry (IHC) and biochemical (DCC) analysis. While the DCC assay showed tumors to be receptor-positive in 62% (ER) and 66% (PR), receptor-positive malignant epithelial cells were only detected in 38% (ER) and 31% (PR) by IHC. There was only a low correlation between the semiquantitative results of ER and PR IHC and the corresponding values of DCC receptor determination. The finding of steroid receptor-positive stromal cells without any evidence of hormone receptor-positive epithelial tumor cells offers a possible explanation for discrepant results in numerous cases with obviously "false positive" results of DCC analysis. Since the considerable heterogeneity of steroid receptor expression present in many ovarian neoplasms can only be detected by IHC, it seems to be the appropriate method of ER and PR determination. Most patients were treated by both radical cytoreductive surgery (n = 76) and a platinum-based chemotherapy (n = 79). ER was not shown to be of significant prognostic value. However, survival was significantly better in patients with PR positive tumors (IHC and DCC) on univariate analysis. Residual tumor after primary surgery was the only remaining significant prognostic factor after multivariate analysis. Further studies are needed to clarify the biological function of steroid receptor-positive stromal cells in ovarian carcinomas.


Detection and patterns of treatment failure in 300 consecutive cases of "early" endometrial cancer after primary surgery.

From November 1977 to July 1987, 300 consecutive patients with endometrial carcinoma clinically confined to the uterine corpus underwent primary surgery consisting of at least abdominal hysterectomy and adnexectomy. Patients with aggressive disease characteristics received postoperative radiotherapy. Forty-seven patients (16%) demonstrated recurrent disease from 2 to 125 (median of 12.8) months after surgery. Forty-seven percent of the recurrences were detected within the first year following surgery and 70% by 2 years after hysterectomy. Of the 47 recurrences, 29 were at distant sites, 16 were within the pelvis, and 2 consisted of both local and distant recurrences. Patients treated with pelvic radiotherapy after hysterectomy were more likely to experience distant, rather than local recurrences. Only 7 of the 148 patients (5%) treated with postoperative radiotherapy recurred in the pelvis. Approximately half of the recurrences were detected in asymptomatic individuals; physical examination and chest X-ray were the most useful means to detect disease in patients without symptoms. The combination of history, physical examination, pap smear, and chest X ray detected all of the recurrences. Actuarial survivals at 12, 24, and 36 months after recurrence were 42, 24, and 17%, respectively. The site of recurrence, time interval of surgery to recurrence, and use of postoperative pelvic radiotherapy were statistically related to patient prognosis. The identification of patients at risk of recurrence and more effective adjuvant therapy need to be developed in order to decrease the frequency of recurrence. In order to substantially improve the survival of patients with recurrent disease, more sensitive methods of detection, as well as more effective salvage therapy, will be required.


Characterization and clinical evaluation of tumor-associated antigen CA54/61 identified by monoclonal antibodies MA54 and MA61 in epithelial ovarian cancer.

Monoclonal antibodies (moABs) MA54 and MA61, directed toward the O-linked mucin-type glycoprotein, have been established and showed highly specific reactivity with human ovarian cancer. Fetal intestinal and colonic mucosal cells expressed this antigen and meconium staining was also frequently positive. To investigate the characteristic of an epitopic carbohydrate recognized by these moABs, the reactivity of each moAB with meconium extract was monitored by solid-phase enzyme-linked immunosorbent assay with mono-, di-, and oligosaccharides. MA54 and MA61 react with meconium extract and the reactivities of these moABs are neuraminidase sensitive. Ovine submaxillary mucin had a strong inhibitory activity toward the reaction between meconium extract and MA54 as well as MA61, suggesting that these moABs recognize NeuAc 2-6GalNAc epitope in meconium. The second aim of this study is to investigate the possible application of moABs to diagnose ovarian cancer and to compare these levels with those of the CA125 antigen. While serum CA54/61 antigen levels were elevated in 44.4% of ovarian cancer cases and serum CA125 antigen levels were elevated in 86.7% of the same population, the use of both assays indicated a sensitivity of detection of 97.8% (44 of 45 patients) in the population studied.


Office laparoscopy and biopsy for evaluation of patients with intraperitoneal carcinomatosis using a new optical catheter.

We report seven patients who underwent diagnostic laparoscopy and biopsy under local anesthesia using a new optical catheter. Six of these procedures were performed in the office. Four patients had previous malignancies (lung, breast, and fallopian tube), and intraperitoneal recurrences or new primaries were suspected. In two of these patients, laparoscopically directed biopsies confirmed adenocarcinoma similar to their prior malignancies, and in one a new primary was diagnosed. The fourth patient had no evidence of intraperitoneal disease. In the three patients with new intraperitoneal malignancies, biopsies obtained laparoscopically confirmed adenocarcinomas of the ovary in two patients and of the gastrointestinal tract in one patient. We feel that this procedure is a safe, simple, effective, and economical way to evaluate the intraperitoneal cavity and to obtain histologic or cytologic specimens for evaluation in patients with intraperitoneal malignancies. This is an ideal, minimally invasive method for detecting small-volume intraperitoneal disease. In addition, it allows some patients to be spared major operative procedures.


The effects of cisplatin on murine metaphase II oocytes.

This study was undertaken to determine if cisplatin can induce aneuploidy in murine oocytes treated prior to ovulation. Control animals were injected with intraperitoneal (ip) saline and the treatment animals were divided into three different dose schedules for ip cisplatin. After ovulation induction, the oocytes were processed for cytogenetic analysis and the chromosomes were counted. There were significantly less oocytes ovulated in the treatment groups when compared to control animals. There was a significant increase in hypohaploid cells counted in the 7.5 mg/kg treatment group, but the percentages of hyperhaploid and polyploid cells were not increased in the treatment groups. The cytogenetic effects of cisplatin in vivo and in vitro are reviewed.


Phase 2 trial of intraperitoneal carboplatin and etoposide as salvage treatment of advanced epithelial ovarian cancer.

To evaluate the efficacy of intraperitoneal (IP) carboplatin-based therapy as salvage treatment of ovarian cancer, 46 patients with persistent or recurrent ovarian cancer following initial systemic chemotherapy were treated with a regimen of carboplatin (200-300 mg/m2) and etoposide (100 mg/m2) administered on a monthly schedule. A maximum of six courses of therapy was delivered, followed by a response laparotomy. The treatment program was well tolerated, except for bone marrow suppression, with one-quarter of patients developing platelet count depressions to < or = 50,000/mm3, and one-third experiencing hemoglobin levels of < or = 8 g/dl during treatment. Twelve (38%) of 32 patients evaluable for efficacy of the treatment program achieved a surgically documented response, including 8 (25%) complete responses. Of 25 patients whose largest tumor mass at the initiation of therapy measured < or = 0.5 cm, 11 (44%) responded, including 8 (32%) complete responses. We conclude that the IP administration of carboplatin can result in surgically documented responses when used in the salvage setting in patients with advanced ovarian cancer. The relative efficacy of carboplatin versus cisplatin when administered by the IP route to patients with ovarian cancer previously treated with platinum-based systemic therapy remains to be defined.


Distribution of intraperitoneally injected microspheres labeled with the alpha-emitter astatine (211At) compared with phosphorus (32P) and yttrium (90Y) colloids in mice.

The alpha-emitter 211At was bound to polymer microspheres with a diameter of 1.8 microns. The distributions in mice of intraperitoneally injected 211At microspheres, 90Y silicate colloid, and 32P chromic phosphate colloid were compared. The microspheres with 211At spread rapidly in the peritoneal cavity and remained mainly on the intraperitoneal surfaces. Intraperitoneal injection of 90Y colloid resulted in high levels in intraperitoneal fat and the diaphragm, but 1 day after injection 8.5% of the injected dose per gram was found in blood and after 6 days 2.5% was observed in bone. The highest accumulation of 32P was found in liver and spleen. The injection of additional nonradioactive chromic phosphate colloid resulted in an even higher accumulation of 32P in spleen and liver. The same phenomenon was not observed with 211At microspheres. It is suggested that it is not only the particle size which is important in the distribution of intraperitoneally injected colloid, but the amount of colloid, the type of colloid, the addition or presence of other substances such as ascites, and the animal species might also influence the distribution. In conclusion, the intraperitoneal distribution of 211At-labeled microspheres in mice was favorable compared with 90Y and 32P colloid. These data must be viewed cautiously since the distribution might be different in other animal species or humans.


Therapeutic efficacy of the alpha-emitter 211At bound on microspheres compared with 90Y and 32P colloids in a murine intraperitoneal tumor model.

alpha-Emitting radionuclides such as 211At have a number of physical characteristics which make them attractive for the treatment of micrometastases. 211At was bound to polymer microspheres and its efficacy was compared with the beta-emitting 32P and 90Y colloids for the treatment of intraperitoneally growing K13 hybridoma tumors in mice. Single graded doses of 0.1-2.5 MBq 211At microspheres injected intraperitoneally 24 hr after inoculation of the hybridoma cells improved survival and produced higher cure rates than 32P colloid, 90Y colloid, or no treatment. One of the most striking contrasts between 211At microspheres and 90Y or 32P colloids was the ability of relatively low doses 211At to affect cures. When comparing the groups with the highest survival rate for each radionuclide (0.1-1 MBq 211At, 2.5 MBq 90Y, and 2.5 MBq 32P), 211At treatment resulted in an improved survival over that with 32P therapy, but the difference was not significant between 211At and 90Y. Toxicity studies with 211At microspheres showed that dosages up to 17 MBq per mouse were not lethal. In conclusion, the present study suggests that the high-energy transfer and the short-range cytotoxicity of the alpha-emitter 211At might be of benefit for intracavitary radiotherapy.


A prospective trial of progesterone therapy for malignant peritoneal cytology in patients with endometrial carcinoma.

From February 1982 to February 1991, 45 patients with endometrial carcinoma confined to the uterus except for malignant peritoneal cytology were treated with 1 year of progesterone therapy. Thirty-six patients have undergone planned second-look laparoscopy with repeat peritoneal washings and the remaining 9 patients either refused second-look laparoscopy or the procedure was medically contraindicated. Of the 36 who underwent second-look laparoscopy, 34 (94.5%) were NED (no evidence of disease) and had negative repeat peritoneal cytology and 2 (5.5%) had persistent malignant cytology. The latter two patients, after an additional year of progesterone therapy, were found to be NED and had negative peritoneal cytology at third-look laparoscopy. Of the 45 women enrolled in this protocol, no patient has developed recurrent endometrial cancer, and the expected 5-year disease-free survival was 88.6%.


Hypogastric artery aneurysm masquerading as an ovarian neoplasm.

Hypogastric artery aneurysm in women is rare. The case presented demonstrates how this lesion can easily mimic an ovarian neoplasm. The missed diagnosis can be catastrophic if the surgeon is unfamiliar with the retroperitoneal anatomy and is confronted with arterial hemorrhage. Hypogastric artery aneurysm should be included in the differential diagnosis of a pelvic mass in elderly women with atherosclerotic disease. The report reviews the literature on the presentation, diagnosis, and recommended treatments.


Synchronous mature teratomas of the ovary and liver: a case presenting 11 years following chemotherapy for immature teratoma.

Immature teratoma is a rare ovarian germ cell tumor with an aggressive clinical behavior. Treatment involves surgical resection, usually followed by chemotherapy. Since the introduction of postoperative chemotherapy several cases of "transformation" of this malignant tumor into mature teratoma (retroconversion) have been described. Usually retroconversion presents during the first year of diagnosis. We report a case of synchronous mature teratomas of the ovary and liver presenting 11 years following chemotherapy for stage III immature ovarian teratoma. The CT scan appearance, incidence, and biological significance of this phenomena are discussed.


Neoadjuvant intraarterial infusion chemotherapy with a combination of mitomycin-C, vincristine, and cisplatin for locally advanced cervical cancer: a preliminary report.

Combination chemotherapy including cisplatin was administered intraarterially from the internal iliac artery as neoadjuvant chemotherapy to six patients with locally advanced uterine cervical cancer (stage higher than IIIB of FIGO). The drugs and doses were mitomycin-C 10 mg/m2, vincristine 1 mg/m2, and cisplatin 50 mg/m2. Two or three courses were repeated at intervals of 3 weeks. In three patients, dose reductions were undertaken for decreased renal function and thrombocytopenia. Partial response was, however, observed in all patients (response rate 100%), and five of six patients were able to undergo a radical hysterectomy. The major toxic effects were leukocytopenia, nausea, and vomiting. Our preliminary experience suggests that pelvic intraarterial infusion of combination chemotherapy is effective against primary and advanced uterine cervical cancer, and this preoperative treatment can lead to easier radical hysterectomy. However, further studies are warranted.


Papillary peritoneal carcinomatosis after prophylactic oophorectomy.

Prophylactic oophorectomy has been recommended in patients with a strongly positive family history for ovarian carcinoma. A patient with a strongly positive family history underwent a prophylactic oophorectomy and, 5 years later, developed a primary peritoneal papillary serous adenocarcinoma. A prophylactic oophorectomy does not afford complete protection in some patients with familial ovarian cancer syndrome. Any tissue derived from the coelomic epithelium may potentially undergo multifocal malignant transformation.


Primary malignant melanoma of the cervix and review of the literature.

Primary malignant melanoma of the cervix was diagnosed in a patient presenting with postmenopausal bleeding. A cervical lesion was biopsied, and diagnosis was established by electron microscopy and staining procedures. After failing radiotherapy, the patient was treated with simple hysterectomy and bilateral salpingo-oophorectomy. The cytology, histology, and electron microscopy of this lesion are presented in addition to a clinical discussion.


[The Ilizarov apparatus in pseudoarthrosis of the long bones].

Distraction osteogenesis with the Ilizarov technique is effective in achieving bone healing in both atrophic and infected pseudoarthrosis. In 10 patients with atrophic and 14 with infected pseudoarthrosis treated by this method, solid bone union with reconstruction of anatomical limb length was achieved in all except 1. They ranged in age from 16-80, with a mean of 44. The mean treatment time was 9.4 months for the femur, 4.5 months for the tibia and 8 months for the radius. Patients could be actively rehabilitated all during the course of treatment and they achieved independent ambulation by the time the apparatus was removed. There were no complications.


[HIV infection in pregnancy].

The epidemic of human immunodeficiency virus (HIV) is now affecting populations that were less susceptible in the early 1980s. It is estimated that 8% of patients with AIDS in the US are women in the reproductive age, and 1% are children. In Israel up to 1992, 134 women and 14 children of infected parents were reported as carrying antibodies against the virus. A pregnant women who was a carrier prompted us to formulate various questions as to how the infection affects pregnancy and vice versa, proper management at delivery, rate and modes of perinatal transmission, risk to health care workers, and legal aspects of HIV infection in Israel.


[Nonsurgical treatment of gallstones].

Lately, nonoperative treatment is being used in high risk patients with gallbladder or bile duct stones. Procedures include oral dissolution, extracorporeal shock wave lithotripsy, mechanical lithotripsy, contact dissolution or extraction after papillotomy or via a T-tube tract. We review nonsurgical treatment of gallstones and present our experience in the treatment of 9 patients using combinations of the above-mentioned modalities.


[Intestinal tuberculosis presenting as Crohn's disease].

A 24-year-old woman who had immigrated from India 3 years before was referred because of diarrhea, abdominal pain and weight loss. Crohn's disease was suspected, but investigation revealed active pulmonary tuberculosis and tuberculosis of the small and large intestine. She was treated with rifampicin, 600 mg/day, INH 300 mg/day, and ethambutol, 400 mg/day, and recovered fully within 6 months.


[Incarcerated obturator hernia causing bowel obstruction].

Obturator hernia is rare and is usually discovered by chance at operation or autopsy. In certain cases obturator hernia may cause sliding and strangulation of abdominal viscera inside the hernial sac, presenting clinically as bowel obstruction. An 83-year-old woman with signs of bowel obstruction was operated after 48 hours of conservative treatment and a small bowel loop was found strangulated inside the sac of an obturator hernia.


[Brain abscess following skull traction].

A case of brain abscess developing after skull traction for fracture of the dens is presented. The abscess was most likely caused by retrograde septic thrombophlebitis via the superficial cortical veins. The abscess was drained and treatment with antibiotics was according to sensitivity on culture. 3 months after surgery there was mild residual paresis of the left hand. The need for close follow-up of patients prone to this complication is emphasized.


Environmental and social determinants of sexual function in the male lesser mouse lemur (Microcebus murinus).

Environmental factors that regulate the sexual activity of male lesser mouse lemurs have been studied experimentally with more than 60 captive animals over an 8-year period. In this nocturnal Malagasy prosimian, variation in day length is the primary factor controlling seasonal sexual activity. Plasma testosterone concentrations were low (= 9 ng/ml) during short days and reached 60 ng/ml during long days (> 12-hour day). This cyclic pattern persists unchanged when artificial photoperiodic rhythms are applied and is not altered by ageing. The timing of puberty is also regulated by photoperiodic changes. Nevertheless, the sexual activity of the male lesser mouse lemur can be dramatically modified by the social environment. In heterosexual groups, behavioural and physiological components of sexual activity are depressed in all males except the dominant one, whose aggressive interactions are always successful. Intermale sexual inhibition was shown to be mediated by chemical cues present in the urine of dominant or isolated males but not in urine of subordinate individuals. The inhibitory signals possess lipophilic properties and are not contingent on the gonadal activity of the urine donor but are linked to adrenocortical activity. By contrast, chemical signals stimulating the reproductive function of all males are found in the urine of females, the presence of which is required for the establishment of clear dominance among grouped males. Endocrine mechanisms underlying intermale sexual inhibition by chemical cues were analysed. Variations in prolactin strongly suggest that olfaction interacts with the photoperiodic regulation of reproductive function, leading to changes in the sensitivity of the negative feedback effect of testosterone on gonadotrophin secretion. Inhibitory or stimulatory effects of chemical signals are discussed in the context of their functional significance for wild populations.


Control of nipple and body contact by mothers and infants in rhesus macaques.

While 3-month-old infant rhesus macaques (Macaca mulatta) were awake and active in social interactions away from their mothers, body and nipple contacts with their mothers were nevertheless made from time to time. In each dyad the proportions of contacts made by the mother nearly equalled those broken by her, suggesting a meshed interaction in which each partner accepted most of the other's contact initiations and terminations. Passive prevention of nipple contact by a mother reduced the frequency of nipple contact by her infant in the first 5 s after the infant had made body contact. Passive prevention occurred after fewer than 1 in 6 body contacts initiated by infants, and--even without its occurrence--most infants were less ready to take the nipple after their own initiatives than after maternal initiatives. Once nipple contact had been made, the probability of breaking body contact was reduced. The role of maternal rejection both in the control of nipple contact in the short term and in determining (through its effect on the sucking pattern) whether the mother gives birth in the next birth season or later is discussed. We suggest that, by the age of 3 months, the infants had already learned when and how often nipple contact with their mothers would be acceptable during their awake and active periods, and we suggest that subsequent decreases in the frequency of nipple contact were partly the results of maternal rejections which were accepted by the infants.


Absorption, distribution and elimination of selenium as L-selenomethionine in non-human primates.

20 adult female macaques (Macaca fascicularis) were given oral doses of L-selenomethionine (L-SeMet) equivalent to 0, 25, 150, 300 and 600 micrograms selenium (Se)/kg body weight, and plasma, erythrocyte, hair, faecal and urine Se concentrations were determined. The macaques were scheduled for 30 daily oral doses of L-SeMet, but systemic toxicity necessitated dose reduction in several animals; two macaques given 600 micrograms Se/kg body weight/day for 10-15 days died, and the concentration of Se in their tissues was determined and compared with Se concentrations in tissues collected from one untreated animal. Circulating and urinary Se concentrations in control macaques were within the normal human ranges. Plasma, erythrocyte, hair and urinary Se concentrations were generally dependent on the dose of L-SeMet administered. Plasma Se reflected more immediately exposure to L-SeMet, whereas erythrocyte Se concentrations increased and decreased more slowly. In some cases, erythrocyte Se was still increasing or showed a plateau after L-SeMet treatment was discontinued. Plasma Se concentrations of 6.7-7.3 ppm were observed in the two animals that died due to acute toxicity to L-SeMet. Neither plasma nor erythrocyte GPx activity was influenced by a single L-SeMet dose, but an increase in erythrocyte GPx activity occurred with continuous exposure. Total tissue Se increased 13-28-fold in macaques given 600 micrograms Se/kg body weight/day for 10-15 days, with the liver and kidneys containing the the highest Se concentrations.


Di-(2-ethylhexyl) phthalate: a short-term repeated inhalation toxicity study including fertility assessment.

In a study of the 28-day inhalation toxicity of di-(2-ethylhexyl) phthalate (DEHP) aerosols, 9-wk-old Wistar rats, 27 males (mean weight 226 g) and 17 females (mean weight 155 g) per group, were exposed in head-nose inhalation systems to DEHP aerosols of respirable particle size (mass median aerodynamic diameter < or = 1.2 microns) or air (controls). Exposure for 6 hr per day, 5 days per wk for 4 wk to target concentrations of 0, 0.01, 0.05 and 1.0 mg/litre gave estimated doses of 230, 11 and 2.3 mg/kg/day for the males, and 360, 18 and 3.6 mg/kg/day for females, on the assumption of 100% deposition and absorption. Clinical investigation and blood chemistry parameters did not reveal any treatment-related effects. At the end of exposure a statistically significant (16%) increase in relative lung weights, accompanied by increased foam-cell proliferation and thickening of the alveolar septi, was found in the males of the highest dose group. Absolute liver weights were significantly (8.75%) increased in females and relative liver weights were increased in both sexes in the highest dose group, but there were no corresponding histological effects. All these effects were reversed during the 8-wk post-exposure period. No testicular toxicity was observed histologically and no impact on mating performance and male fertility was detected after two matings of treated males with untreated females, 2 and 6 wk after the end of exposure. Electron microscopic examination of liver samples from two male and two female rats per group at the end of exposure and after the 8-wk post-exposure period did not reveal clear substructural changes that could be attributed to exposure or to peroxisome proliferation. The no-observed-effect level for all exposure-related findings was 0.05 mg/litre under the conditions used.


Influence of intact and myrosinase-treated indolyl glucosinolates on the metabolism in vivo of metronidazole and antipyrine in the rat.

Induction of the cytochrome P-450 enzymes is a mechanism whereby cruciferous vegetables and their glucosinolates could influence the risk of cancer. The cytochrome P-450-inducing capacity of isolated intact broccoli glucosinolates and their degradation products, resulting from myrosinase-catalysed hydrolysis, has been assessed in studies of the metabolism of antipyrine (AP) and metronidazole (MZ) in the rat. The intact glucosinolates had no effect on the metabolism of MZ and AP as measured by the clearance and metabolite formation rates; however, the myrosinase-treated glucosinolates significantly increased the clearance of AP by two-thirds and the formation rates of the three major AP metabolites by 87-100%, and doubled the rate of oxidative metabolism of MZ to its hydroxy and acetic acid metabolites. Active myrosinase was thus essential for the capacity of glucosinolates from broccoli (mainly indolyl glucosinolates) to induce the activity of several cytochrome P-450 isoenzymes involved in the metabolism of AP and MZ. The data indicated that hydrolysis products of indolyl glucosinolates had an inducing effect on the activity, but not the total amount, of hepatic cytochrome P-450 isoenzymes. The effect of these products on the oxidative metabolism of AP and MZ was similar to that of phenobarbital. The significance of this induction pattern in relation to cancer risk depends primarily on the activation/inactivation mechanism of the relevant carcinogen.


Comparison of the effects of nafenopin on hepatic peroxisome proliferation and replicative DNA synthesis in the rat and Syrian hamster.

Male Sprague-Dawley rats were fed control or 0.1% nafenopin diet and male Syrian hamsters were fed control or 0.25% nafenopin diet for periods of 7 and 54 days. Nafenopin treatment produced a sustained increase in liver weight and induction of hepatic peroxisomal and microsomal fatty acid-oxidizing enzyme activities, with a greater effect being observed in the rat. Replicative DNA synthesis was studied by implanting osmotic pumps containing [3H]thymidine during study days 0-7 and 47-54. Cell replication, determined either as the hepatocyte labelling index or by incorporation of radioactivity into liver whole homogenate DNA, was increased in rats given nafenopin for 7 and 54 days. In contrast to the rat, no significant effect on replicative DNA synthesis was observed in the Syrian hamster. These results provide further evidence for species differences in hepatic peroxisome proliferation, with the Syrian hamster being less responsive than the rat. Furthermore, while peroxisome proliferators produce hyperplasia in rat and mouse liver, these data suggest that they may not have any marked effect on hepatic replicative DNA synthesis in the Syrian hamster.


Teratism induced in the developing chick by RPR-V, an organophosphate.

Fertile White Leghorn chicken eggs were injected on day 4 of incubation with 0, 0.25, 0.50 or 1.00 mg RPR-V/egg and opened on day 20 or allowed to hatch. The following parameters were examined: hatchability, growth and development, external malformations and skeletal deformities. As the dose was increased, the hatchability decreased, and the incidence of deformities increased. Staining of the skeleton with Alizarin red showed clear deformities in the embryos. The results suggest that RPR-V has teratogenic effects on chick embryos when injected on day 4 of incubation.


Anticarcinogenic effects of some Indian plant products.

The anticarcinogenic properties of some commonly consumed spices and leafy vegetables were investigated. The effects of feeding the plant products on the induction of squamous cell carcinomas in the stomachs of Swiss mice by feeding benzo[a]pyrene(B[a]P) and on the induction of hepatomas in Wistar rats by feeding 3'-methyl-4-dimethylaminoazobenzene (3'MeDAB) were investigated. Among the nine plant products tested, cumin seeds (Cuminum cyminum Linn) and basil leaves (Ocimum sanctum Linn) significantly decreased the incidence of both B[a]P-induced neoplasia and 3'MeDAB-induced hepatomas. Poppy seeds (Papaver somniferum Linn) significantly inhibited B[a]P-induced neoplasia alone, while the other plant products, asafoetida, kandathipili, turmeric, drumstick leaves, solanum leaves and alternanthera leaves were ineffective. These results suggest that cumin seeds, basil leaves and to a lesser extent poppy seeds, which are all widely used in Indian cooking, may prove to be valuable anticarcinogenic agents.


Chronic oral toxicity and carcinogenicity study of stevioside in rats.

Groups of 45 male and 45 female inbred Wistar rats were given diets containing stevioside (85% pure) at 0, 0.2, 0.6 or 1.2% for 2 yr. After 6, 12 and 24 months, five rats from each group were killed for haematological and clinical biochemical tests. Growth, food utilization and consumption, general appearance and mortality were similar in treated and control groups. The mean lifespan of rats given stevioside was not significantly different from that of the controls. No treatment-related changes were observed in haematological, urinary or clinical biochemical values at any stage of the study. The incidence and severity of non-neoplastic and neoplastic changes were unrelated to the level of stevioside in the diet. The maximum no-observed-effect level of stevioside was 1.2%, and an acceptable daily intake of stevioside for humans of 7.938 mg/kg body weight/day is suggested.


Effect of extraneous supplementation of ascorbic acid on the bio-disposition of benzanthrone in guinea pigs.

The bio-elimination and organ retention of orally administered [14C]benzanthrone, an anthraquinone dye intermediate, were determined in control and ascorbic acid-supplemented guinea pigs. Urinary excretion of benzanthrone in control and ascorbic acid-treated animals during 96 hr was 27.9 and 30.5%, respectively, with peak elimination at 48 hr. Faecal elimination in control and supplemented animals during 96 hr was 24.5 and 38.8%, respectively, with a peak at 48 hr. The organ retention of radiolabelled benzanthrone at the end of 96 hr was of the order of 39% in control animals (gastro-intestinal tract 16%; liver 22%; testis 1.2%); ascorbic acid supplementation reduced benzanthrone retention to 19.5% (gastro-intestinal tract 12.7%; liver 6.8%). Overall, pretreatment of guinea pigs with ascorbic acid caused a 32% enhancement in the clearance of radiolabelled benzanthrone through the urine and faeces, while organ retention was reduced by about 50%. A prophylactic dose of ascorbic acid may prevent benzanthrone-induced toxic symptoms in exposed workers.


Liver medium-term bioassay in rats for screening of carcinogens and modifying factors in hepatocarcinogenesis.

The present report describes a study of the hepatocarcinogenic potential of a second large assay series of 94 compounds carried out using the rapid bioassay system (DEN-PH model) developed in this laboratory and based on the two-step concept of hepatocarcinogenesis. Male F344 rats were initially given a single dose of diethylnitrosamine (DEN, 200 mg/kg body weight ip) and, starting 2 wk later, were treated with test compounds for 6 wk and then killed, all rats being subjected to a two-thirds partial hepatectomy at wk 3. Carcinogenic potential was scored by comparing the numbers (no./cm2) and areas (mm2/cm2) of induced glutathione S-transferase placental form (GST-P) positive foci in the livers of groups of about 15 rats with those of corresponding control groups given DEN alone. Positive was scored for a significant increase (P < 0.05) in quantitative values of GST-P positive foci, negative for no change or a decrease. Results for the 94 compounds were also compared with previously published data from Salmonella/microsome (Ames) tests and long-term carcinogenicity studies in rats and mice. Of the known liver carcinogens, 14 out of 14 (100%) mutagenic (Ames test) compounds and 10 out of 12 (83%) non-mutagenic compounds gave positive results in our DEN-PH system (mean 92%). Two hepatocarcinogenic peroxisome proliferators did not enhance the development of GST-P positive foci. Carcinogens other than hepatocarcinogens gave fewer positive results (five out of 17, 29%). One of the 13 compounds reported as non-carcinogenic, malathion, gave positive results in the DEN-PH assay, suggesting that this compound is a weak hepatocarcinogen or tumour promoter for hepatocarcinogenesis based on the two-stage hypothesis for carcinogenesis. The present study also provided information regarding the inhibitory potential of nine compounds. The practical usefulness and benefits of the DEN-PH protocol for the rapid screening of carcinogenic agents are discussed.


Aflatoxins in cosmetics containing substrates for aflatoxin-producing fungi.

29 random samples of eight different products of skin cleansing creams or powders and skin peeling products (including practically all products of this type available on the German market), containing substrates for aflatoxin-producing fungi or plant parts directly connected with such substrates, such as almond bran, peach bran, apricot seeds, wheat bran, horse chestnut and cob were analysed for aflatoxin B1, B2, G1 and G2. In 11 (38%) of the samples aflatoxins were not detectable ( < 0.1 micrograms/kg); 18 (62%) of the samples contained from 0.4 to 78.5 micrograms aflatoxin/kg. The majority of the positive samples contained all four types of aflatoxin. Aflatoxins can be absorbed through the skin. To what extent this happens with these products and the resulting toxicological significance is difficult to estimate. Nevertheless, potent carcinogens such as aflatoxins should be completely absent from cosmetics; this can be achieved by careful quality control of the raw materials.


Role of gastric glutathione in smoke flavouring-induced gastric injury in rats.

Some commercial liquid smoke flavourings have been shown to induce acute gastric mucosal injury in rats when given orally as a large single dose. The present study was carried out to examine the mechanism of action in rats of two selected smoke flavourings containing about 10% total acids as acetic acid. These flavourings and 10% acetic acid decreased the concentration of glutathione (GSH) in the glandular stomach. The decrease in gastric GSH was coupled with smoke flavouring-induced gastric injury. Pretreatment with N-ethylmaleimide, a GSH depletor, enhanced acetic acid-induced gastric injury. Pretreatment with cysteine, a sulphhydryl compound, protected rats against smoke flavouring-induced gastric injury. Aqueous fractions of the smoke flavourings, after removal of non-polar compounds and acidic organic compounds (including acetic acid) by diethyl ether extraction, decreased the gastric GSH concentration considerably and had a marked reactivity in vitro with GSH, but these fractions by themselves showed no ability to induce gastric injury. Addition of 10% acetic acid to these aqueous fractions caused greater gastric injury than 10% acetic acid alone, which suggests that these aqueous fractions contain the (unidentified) compound(s) that facilitate acetic acid-induced gastric injury. These findings indicate that gastric endogenous and exogenous sulphhydryls play an important part in gastric cytoprotection.


The influence of ascorbic acid on lipid peroxidation in guinea pigs intoxicated with cadmium.

Cadmium (as CdCl2) administered at a dose of 1 mg Cd/animal/day in drinking water to guinea pigs with a low intake of ascorbic acid (2 mg/animal/day) increased lipid peroxidation (LPO) in the kidney, liver and serum after 5 wk of treatment. A high intake of ascorbic acid (100 mg/animal/day) decreased LPO (determined from malondialdehyde levels) in the kidney, liver and serum of Cd-treated guinea pigs. Administration of Cd to guinea pigs with a low intake of ascorbic acid significantly increased LPO in the kidney at 5 wk, in comparison with controls given low levels of ascorbic acid but no Cd. However, at 12 wk, LPO in the kidney of Cd-treated guinea pigs was significantly lower than in the controls. These results indicate that the level of lipid peroxides in the kidney of guinea pigs with a low intake of ascorbic acid depends on the amount of accumulated cadmium.


Reproductive and neurobehavioural effects in three-generation toxicity study of piperonyl butoxide administered to mice.

Piperonyl butoxide (PB) was administered continuously to mice from 5 weeks of age in the F0 generation to weaning of the F2 generation. PB was administered in the diet at levels of 0 (control), 0.1, 0.2, 0.4 and 0.8%. Selected reproductive, developmental and behavioural parameters were measured. Litter size and litter weight were reduced in higher-dosed groups, and the body weight of the pups in the lactation period was reduced in dosed pups in each generation. The survival index at postnatal day 21 of the group receiving 0.8% PB was reduced in each generation. The developmental and behavioural parameters in the lactation period were little different from those of the controls, apart from olfactory orientation in the F1 generation. However, in the F2 generation mice, surface righting, cliff avoidance and olfactory orientation were adversely affected in treatment groups. The results suggest that PB had adverse effects on reproductive, developmental and behavioural parameters of mice, with increasing effects in subsequent generations of offspring.


Acute renal toxicity of thiabendazole (TBZ) in ICR mice.

The acute toxic effects of thiabendazole [2-(4'-thiazolyl)benzimidazole; TBZ] on the kidneys of ICR mice were investigated. The mice were given 0, 250, 500 or 1000 mg TBZ/kg body weight by gavage (using olive oil as a vehicle), and the kidneys were subjected to pathological examination at 1, 3, 5 or 24 hr after dosing. Gross findings were slight enlargement and the presence of whitish areas (white maculae) in kidneys of treated mice at 3, 5 or 24 hr after dosing. Histological findings were desquamation of degenerated cells in proximal tubules of treated mice at 1 hr. Dilation of proximal, distal and collecting tubules was apparent in treated mice at 3, 5 and 24 hr. TBZ-induced renal injury was reduced by pretreatment with inducers of the microsomal monooxygenase system (sodium phenobarbital, beta-naphthoflavone and 3-methylcholanthrene) and were enhanced by pretreatment with inhibitors of that system (2-diethylaminoethyl-2,2-diphenylvalerate hydrochloride and piperonyl butoxide). The concentration of TBZ in blood at 1 or 5 hr after dosing was lower in mice pretreated with microsomal monooxygenase system inducers and was higher in those pretreated with the inhibitors, than in those given TBZ alone. These results suggest that TBZ-induced renal injury may be attributable to the parent compound rather than its metabolites. Measurement of organic ion uptake into renal slices revealed significant depression of [1-14C]tetraethylammonium bromide (TEA) uptake in treated mice at 1 or 5 hr, whereas uptake of p-[glycyl-2-3H]aminohippurate (PAH) was not depressed at 1 or 5 hr after dosing. The reduction in uptake of TEA is interpreted as the result of competitive suppression of the tubular transport of TEA by TBZ. TBZ-induced renal injury was reduced by organic cation transport inhibitors [N'-methylnicotinamide (NMN) or thiamine] but not by organic anion transport inhibitor [p-(dipropylsulphamyl)benzoic acid probenecid], suggesting that the reduction of TBZ-induced renal injury is the result of competitive suppression of the tubular transport of TBZ by NMN or thiamine.


Toxicology and hazard potential of trifluralin.

This paper reviews the results of toxicity studies conducted in laboratory animals to evaluate the safety of the herbicide trifluralin (TFL). The data show that TFL is slightly toxic following single oral exposure. Testing for embryotoxicity in rats and rabbits indicated no teratogenic potential, and many different mutagenicity tests showed that TFL was non-genotoxic. Subchronic and chronic toxicity testing in rats, mice and dogs indicated that TFL was haematotoxic (anaemia and methaemoglobinaemia), particularly in the dog, and slightly hepatotoxic. No-observed-effect levels of 4.8 and 41 mg/kg body weight/day, respectively, were determined in dogs and rats exposed chronically to TFL. Oncogenicity studies in rats and mice revealed no carcinogenic potential. Since the data for TFL indicated no mutagenic or other special toxicological risks, it is suggested that a safety factor of 100 could be used for the determination of the acceptable daily intake of TFL, which would be 0.05 mg/kg body weight/day.


Genotoxicity of 'gudakhu', a tobacco preparation. II. In habitual users.

The genotoxic potential of 'gudakhu', a paste-like tobacco preparation that is used widely in Orissa, India, was evaluated using the micronucleus test in exfoliated cells of the buccal mucosa. Cells from 120 habitual users and from 102 non-users were examined. The incidence of micronuclei (MN) was increased in the mucosa cells of users, and the increase was significant in those who had used gudakhu for more than 5 yr. The increased incidence of MN was significantly correlated with the period of use of gudakhu, as well as with the frequency of daily use. There were no significant differences between the results for men and women.


Evaluation of the cumulative (repeated application) eye irritation and corneal staining potential of FD&C yellow no. 5, FD&C blue no. 1 and FD&C blue no. 1 aluminium lake.

A protocol to evaluate the ocular irritation, staining, and embedding potential of FD&C colours (Yellow No. 5, Blue No. 1, Blue No. 1 Aluminium Lake) produced by repeated topical application to rabbit eyes is described. Test materials (3%, w/v in aqueous vehicle) were administered once daily, for a total of 21 days, to the conjunctival sac of the right eye of New Zealand White Rabbits (6 of each sex per group) at a dose volume of 30 microliters. Control animals (6 of each sex) received 30 microliters of the vehicle daily. All animals survived and were free of significant clinical signs of toxicity throughout the study. Ophthalmoscopic examinations revealed that all animals were free of abnormalities considered to be of clinical importance; all animals were free of significant signs of ocular irritation, staining and particle embedment. The results of this study support the safe use of these materials in consumer products intended for use in the eye area.


Analysis of alternative methods for determining ocular irritation.

According to classification and labelling requirements, chemicals, dyes, agrochemicals and pharmaceutical formulations have to be evaluated for their potential to induce eye irritancy or corrosion. An attempt was made to analyse the predictive power of the bovine eye-chicken egg chorioallantoic membrane (BE-CAM) assay in comparison with results obtained using the conventional Draize method. In summary, results showed limited correlation between reactions in vitro and responses of eyes in vivo. In a pilot study, ultrasonic pachymetry showed high sensitivity and fairly good correlation between corneal thickness and clinical observations in eyes.


Toxicokinetics of cinnamaldehyde in F344 rats.

The toxicokinetic profile of cinnamaldehyde (CNMA) was investigated in Fischer 344 rats. CNMA was found to be unstable in blood. After iv administration, a large fraction of CNMA was immediately oxidized to cinnamic acid. The biological half-life of CNMA after iv administration was found to be 1.7 hr. After administration by gavage of CNMA at 250 or 500 mg/kg body weight using corn oil as vehicle, the maximum blood concentrations of CNMA were in the order of 1 microgram/ml. These low blood concentrations were maintained over a 24-hr period after a dose of 500 mg/kg, which is relatively long considering the short (1.7 hr) biological half-life of CNMA. The estimated oral bioavailability of CNMA was less than 20% for both the 250 and 500 mg/kg doses. No CNMA was present in blood at any time in rats dosed with 50 mg CNMA/kg body weight. Only a small amount of the administered CNMA was excreted in rat urine as free cinnamic acid or beta-glucuronide-conjugated cinnamic acid. The majority of CNMA administered orally was excreted in urine as hippuric acid within 24 hr. The maximum excretion rate occurred at 8 hr after gavage. Hippuric acid recovered in 50-hr urine samples was found to be directly proportional to the oral dose of CNMA.


Sex determination by discriminant function analysis of lateral cranial form.

To sex the cranium, morphological features of cranial specimens were quantified with a personal computer that automatically measures distance and gradient for 39 craniometric points in the lateral contour line of the skull, which were digitized by a tablet digitizer connected to the computer. Specimens used for discriminant analysis were 50 male and 50 female adult Japanese skulls. The lateral contour showed sex differences in the nasal bone, supraorbital ridge, forehead and vertex. The nasal bone and supraorbital ridge were more developed in male contour line, and the forehead was more rounded in female contour line. But compared with the supraorbital ridge and forehead, the vertex had a wide variety of contour lines in both sexes. The vertex seemed to be less reliable as the indicator of sex. The sex differences were better reflected by gradient than distance. From variables of the gradient and distance showing significant sex differences, the discriminant function was derived and tested in 21 other specimens (13 male and 8 female skulls). The mean ratio of correct sexing of the human skull by the discriminant function was 86%.


Chemiluminescent DNA probes: evaluation and usefulness in forensic cases.

Five phosphatase-labelled oligonucleotide probes were evaluated in respect to their sensitivity, with the help of an optimized chemiluminescent protocol, for DNA-VNTR polymorphism determination. Their usefulness for the identification of biological traces is illustrated with casework examples.


A pathological study of sudden coronary death in China: report of 89 autopsy cases.

This paper reports the results of a forensic pathological study of 89 autopsy cases of Sudden Coronary Death (SCD). Of 89 cases, 63 (52 male, 11 female) were narrowed by 76-100% in cross-sectional area (XSA) of the coronary artery (CA) and 26 (22 male, 4 female), by 51-75%. Atherosclerotic plaques in the CA were serious and extensive, especially in the left anterior descending and often involved several branches of CA at the same time. Recent thrombosis was found in 18 cases, haemorrhage in plaques in 17 cases. Only 2 cases had visible acute myocardial infarction. Inflammatory cell infiltration was found in coronary plaques in 36 cases. Myocardial fibrosis or small scar formation was detected in 51 cases. It is suggested that although the incidence of coronary heart disease (CHD) is low in China, SCD is the commonest mode of Sudden Unexpected Death. The majority of SCD (52%) were middle aged males (30-49 years old). Most of the cases died suddenly during sleep without any clear precipitating factors. The characteristics of occurrence and pathological changes in the CA and myocardium and the pathological diagnosis of SCD are also analysed and discussed.


Detection of human proteins in buried blood using ELISA and monoclonal antibodies: towards the reliable species indentification of blood stains on buried material.

The survival of human proteins in blood stains on fragments of cloth buried in exposed soil was examined in a 15-month investigation carried out from September 1990 to December 1991. During this period there was a wide variety of weather conditions. Samples were exhumed at 4-weekly intervals for 16 weeks and finally at 65 weeks; extracts of the stains were tested for albumin and IgG using a highly specific and sensitive enzyme-linked immunosorbent assay (ELISA) performed with monoclonal antibodies. Human albumin survived well throughout the 15 months of study, but IgG could be detected only in the 4- and 8-week samples. The reactions for IgG were weaker than those for albumin, although the method's sensitivity (10 ng) was the same for each protein. Appropriate buried and non-buried control experiments were carried out using cloth, either unstained or stained with human blood or animal sera; there was no cross-reactivity between human and the other species investigated and soil did not affect the assay; under laboratory conditions, IgG and albumin survived equally well. The system's versatility was illustrated by using monoclonal anti-bovine-albumin to detect specific albumin in the extracts of buried cloth which has been stained with bovine serum. It was concluded that ELISA performed with monoclonal antibodies could be of great value in identifying blood stains for forensic purposes.(ABSTRACT TRUNCATED AT 250 WORDS)


Dissecting intramural haematoma of the coronary artery with intimal tears.

A dissecting intramural haematoma of the coronary artery is an infrequent cause of sudden and unexpected death. Most cases are women and in a significant number of these women, the haematoma occurred at term to 80 days postpartum. Because the intimal rupture and communication with the lumen were not found in most of the cases, the site of rupture has been considered to be a vasa vasorum. The authors report on a case of one woman who was at a later postpartum period (106 days). The microscopic finding of the dissected coronary artery revealed two intimal tears in a section. The case showed that the dissecting intramural haematoma of the coronary artery is caused not only by the rupture of the vasa vasorum but also by the rupture of the intima.


Evaluating suspected co-proxamol overdose.

In three instances of suicidal poisoning by co-proxamol (paracetamol and dextropropoxyphene) blood samples were obtained from 11 sites together with eight tissue samples, bile, urine, gastric contents and duodenal contents. Site-dependent differences in blood propoxyphene concentration varied between the three cases but concentrations were consistently lowest in peripheral blood and highest in central sites: 3.9-5.5 (pulmonary vein) mg/l; 4.6-25 (pulmonary vein) mg/l; 3.2-40 (aorta) mg/l. There was a less than twofold variation in corresponding blood paracetamol concentrations. Reference data on fatal propoxyphene blood concentrations do not specify the blood sampling site and can be misleading. The intra-individual variability of propoxyphene concentrations in blood in these three cases underscores this problem. Tissue concentrations of propoxyphene showed considerable inter-individual variability in degree and pattern. Tissue concentrations of paracetamol showed a less than twofold intra-individual variation. Body drug loads were calculated by two methods: from organ weights and tissue concentrations; from published volume of distribution data (Vd). For paracetamol the body drug load is underestimated by the organ weight calculation but the Vd calculation approximates the suspected dose based on anamnestic information. For propoxyphene the body drug load is seriously underestimated by the organ weight calculation and overestimated up to 2.5 times by the Vd calculation. Since the two drugs have a fixed ratio in co-proxamol then the dose of propoxyphene (the effective lethal agent) can be inferred from the paracetamol dose calculated by Vd. This approach may be applicable to cases of overdose with other compounded drug preparations.


Sudden death associated with solitary intracavitary right atrial metastatic tumour deposit.

We report a case of cardiac metastasis of primary adrenal cortical carcinoma to the endocardium of the right atrium resulting in an intracavitary mass which we believe caused sudden death by blocking the tricuspid valve orifice. We have been unable to find a similar case previously reported although primary atrial myxoma is well known to be a rare cause of sudden cardiac death by the same mechanism. Modern non-invasive imaging techniques have facilitated recognition of intracavitary cardiac metastases, and with surgical intervention, when feasible, the risk of sudden cardiac death can be reduced.


Firearm deaths in Yorkshire and Humberside.

Firearm fatalities occurring in Yorkshire and Humberside between 1975-1991 were examined; 130 cases were found, 64 were homicides, 55 suicides and 11 were accidental or unclassifiable. All but two of the suicides were men. Shotguns were the most frequent weapon used in both suicides and homicides. Of the 55 suicides, 17 were by men who had murdered before committing suicide. In 15 of these cases the assailants knew their victims. The other two cases involved murder of police officers. These cases were mostly with shotguns. In both homicide and suicide, rifled weapons and handguns were infrequently used. Most suicides and homicides occurred at home. Alcohol was again shown to be a significant factor in suicides. Overall firearms account for less than 10% of homicides in England and Wales and less than 5% of suicides in men and is most unusual in women.


Comparing roadside with subsequent breath alcohol analyses and their relevance to the issue of retrograde extrapolation.

Driving while intoxicated (DWI) legislation requires proving the critical breath alcohol concentration (BrAC) at the time of driving. With time delayed analysis, retrograde extrapolation is occasionally employed but has several uncertainties associated with it. The present study attempts to address whether subjects actually arrested for DWI are likely to have BrAC values near the time of driving differing largely from those performed at a subsequent time. Selected officers arrested n = 161 subjects where roadside BrAC was determined with Pre-Arrest Breath Test (PBT) devices along with subsequent duplicate evidential analyses followed by an additional PBT analysis. These two sets of duplicates, one with large time interval (mean = 63.5 min.) and one with a 2-3 min difference, were then compared by several statistical methods. The results showing duplicate variability did not differ when the long time interval existed (F = 1.0, P > 0.05). A small but significant decrease in BrAC with respect to time appeared for the duplicate PBT data. Retrograde extrapolation applied to the data employing an assumed 0.015 g/210 l/h yielded a small but significant overestimate of the actual roadside PBT result. Finally, evidentiary analyses performed within 2 h of driving will provide good estimates and certainly not overestimates, of the BrAC existing at the time of driving and it appears that extrapolation may be unwarranted in these cases.


Gas chromatographic detection of cocaine and cocaethylene in hair of mice chronically injected with cocaine or cocaethylene and fed ethanol.

GC and GC/MS analysis was used to detect cocaine and cocaethylene in hair extracts of mice injected with 20 mg/kg cocaine hydrochloride or an equivalent dose of cocaethylene fumarate twice daily for 3 weeks. Some mice were fed liquid Lieber-DeCarli diets containing ethanol (26% of total calories) and injected twice daily with the same doses of cocaine or cocaethylene or combination of cocaine and morphine (5 mg/kg). The average concentrations of cocaine in different experimental groups were in the range of 0.9-2.4 ng/mg of hair and for cocaethylene, 2.4-2.8 ng/mg of hair. There were no significant differences in hair concentrations of cocaine among groups receiving cocaine treatment, nor were there significant difference in cocaethylene concentration in hair in the two groups administered cocaethylene. In hair extracts of mice treated with cocaine and ethanol, levels of cocaethylene were below the limit of detection.


Lipid oxidation and atherosclerosis.

Hypercholesterolemia, and in particular high levels of low density lipoprotein (LDL), is a well established risk factor for development of coronary heart disease (CHD), but the biological mechanisms by which LDL promote formation of atherosclerotic plaques are still poorly understood. During the last decade several lines of evidence have suggested that oxidative modification of LDL is a key process in this respect. Oxidation of LDL has been found to increase its uptake in macrophages and lead to formation of macrophage foam cells. Other studies have indicated that oxidized LDL may induce vascular inflammation and even give rise to autoimmune reactions in the vascular wall. These findings have it made important to investigate the possible role of LDL oxidation in CHD also in clinical studies and the initial results of such studies support the notion that oxidation of LDL also may be of clinical relevance. In a group of young post-myocardial infarction patients the in vitro susceptibility of LDL to oxidative modification was found to be significantly related to the severity of coronary atherosclerosis as assessed by angiography. In another study presence of antibodies against oxidized LDL was found to be associated with increased progression of carotid disease. Should these findings be confirmed in larger patient groups and LDL oxidation established as a key factor in the development of atherosclerosis this would have a considerable impact on future strategies for prevention and treatment of coronary heart disease.


Gene expression in atherosclerotic lesions.

Atherosclerotic lesions contain macrophages, smooth muscle cells and endothelial cells, all of which participate in lesion development. Upon stimulation these cells express a variety of different factors and receptors which are involved in lipid metabolism, cellular proliferation, tissue repair, immune response and inflammatory reactions. During the last few years, active expression of several genes has been reported in developing atherosclerotic lesions. These genes include scavenger receptor, 15-lipoxygenase, monocyte chemoattractant protein-1, macrophage colony stimulating factor-1, lipoprotein lipase, platelet-derived growth factor, tissue factor, apolipoprotein E, stromelysin, different adhesion molecules and various cytokines. Evidence continues to grow that the pattern of gene expression and the functional status of cells in different regions of atherosclerotic lesions may differ considerably and could be regulated by local factors present in atherosclerotic lesions. One such a powerful factor which may contribute to the regulation of gene expression in developing lesions is oxidized LDL which has been shown to affect the expression of cytokines, growth factors and chemotactic factors by arterial cells. Recent developments in the analysis of gene expression in the artery wall at the cellular level will undoubtedly increase our understanding about the sequence of events leading to the formation of atherosclerotic lesions.


[Role of smooth muscle cell proliferation after balloon angioplasty].

The long-term success of percutaneous transluminal coronary angioplasty (PTCA) is severely limited by the high incidence of restenosis, which occurs in up to 30 to 40% of primary successfully treated patients. Several postmortem and experimental studies have demonstrated that restenosis after balloon angioplasty is caused by intimal proliferation of smooth muscle cells (SMCs). As a result of the vessel wall injury due to the mechanical intervention, endothelial injury and platelet aggregation at the site of dilatation can be observed. SMCs are activated for migration and proliferation in the early phase after angioplasty by subsequent expression of several growth factors, activation of macrophages, and expression of a variety of mitogens (Figure 1). Since several other alternative interventional devices like atherectomy, excimer laser angioplasty and stenting are also limited by the occurrence of SMC proliferation following treatment, there is an obvious need for a pharmacological approach to inhibit SMC proliferation after PTCA. It has been shown in experimental studies that the proliferative response of the vessel wall in the intima occurs within the first seven days after dilatation. A significant increased mitosis rate of SMCs in the media, however, can be observed within three weeks after intervention (Figure 2). As suggested by several authors, macrophages are thought to play an important role in the restenosis process after balloon angioplasty. However, our experimental results demonstrate that no significant accumulation of macrophages occurred prior to seven days following balloon angioplasty (Figure 3).(ABSTRACT TRUNCATED AT 250 WORDS)


Atherosclerosis--where are we heading?

Aim of this review is, to analyse the current knowledge of the pathogenesis of atherosclerosis from the viewpoint of a clinical cardiologists. All the current ideas about the pathogenesis of atherosclerosis have been derived from the theories of three pathologists. The first was Rokitanski 1852, his theory was somewhat modified by Virchow 1856 and revitalised and modified again by Ross in 1986. All current theories however are based on the term response to injury leading to accelerated atherosclerosis. Following the classification of Stary et al. and the suggestions of the report of the Committee on Vascular Lesions of the Council of Arteriosclerosis the early lesions (Stary class I and II) should be renamed as normal vascular adaptation to physical or chemical causes. These causes include not only shears stress or turbulences due to increased blood pressure but also hypercholesterolemia and certain changes of plasma protein content. The classification of Fuster et al. is mainly dependent on a mechanical injury to the arterial wall induced by the interventional cardiologist. Thus far, it seems questionable, if such a response to injury is indeed the leading mechanism ultimately resulting in advanced arteriosclerosis. In view of the multifactorial processes involved in the pathogenesis of either adaptation or then accelerated arteriosclerosis we have to admit, that our insight is still limited. Thus far, the only lesson to be learned for practising clinicians is that we have to accept that the process of normal adaptation of the vascular arterial wall to the abnormal development of a clinical significant lesion is dynamic and is not well characterized by the response-to-injury theory. We do know that even in the stage of acute coronary syndromes lesion changes are unpredictable. This coincides with the finding from the experimental laboratory that lesion growth is episodic and a result of different stages of activity of the involved cell types and growth factors. Thus, we should avoid looking only for certain stages but rather accept that this is 1. a normal adaptative alteration of the arterial wall which might 2. result in accelerated atherosclerosis due to genetic determination of smooth muscle cells and accumulation of a variety of growth factors.


Self-heated thermistor measurements of perfusion.

A microcomputer-based control system applies a combination of steady state and sinusoidal power to a thermistor probe which is inserted into the tissue of interest. The steady-state temperature response is an indication of the effective thermal conductivity (keff), which includes a component due to intrinsic conduction plus a convective component due to the tissue blood flow near the probe. By careful choice of the excitation frequency, the sinusoidal temperature response can be used to measure intrinsic thermal conductivity (km) in the presence of blood flow. Optimal sinusoidal heating frequency depends on the thermistor size. Experimental results in the alcohol-fixed canine kidney cortex show that perfusion is linearly related to the difference keff minus km. The instrument can measure tissue thermal conductivity with an accuracy of 2%. The instrument can resolve changes in perfusion of 10 mL/100g-min with a Thermometrics P60DA102M thermistor. The maximum error in measured perfusion is about 30%. When tissue trauma due to probe insertion is minimized, the self-heated thermistor method gives a reliable indication of local tissue blood flow.


Multifiber, multiwavelength, fiber optic fluorescence spectrophotometer.

A fiberoptic spectrofluorimeter was constructed capable of remote operation of up to 18 biosensors under fully automated, multiplexed control. Excitation and emission wavelengths were separately adjustable, as were optical bandwidths, signal gain, phase, integration time, and illumination duty cycle. Synchronous demodulation was used to enhance signal-to-noise ratio and allow operation under ambient lighting conditions. Detection limit for aqueous sodium-fluorescein was below 1 ng/mL at greater than unity signal-to-noise ratio, with effective sampling volumes less than 0.02 microL. The fluorimeter showed characteristics fiberoptic sampling behavior with asymptotic nonzero signal magnitudes under increasing pathlength conditions.


Regeneration microelectrode array for peripheral nerve recording and stimulation.

A microelectrode array capable of recording from and stimulating peripheral nerves at prolonged intervals after surgical implantation has been demonstrated. The microelectrode array, fabricated on a silicon substrate perforated by multiple holes (referred to as via holes), is implanted between the ends of a surgically severed nerve. Regenerating tissue fixes the device in place to provide a stable mapping between the microelectrodes and the axons in the nerve. Processes were developed for the fabrication of thin-film iridium microelectrodes, micromachined via holes, and silicon nitride passivation layers. All fabrication methods were designed to be compatible with standard CMOS/BiCMOS processes to allow for on-chip signal processing circuits in future designs. Such arrays, implanted in the peroneal nerves of rats, were used to record from and stimulate the nerves at up to 13 months postoperatively.