Analgesic effect of salmon-calcitonin administered by two routes. Effect on morphine analgesia. We compared the analgesia induced by intraperitoneally (i.p.) and intracerebroventricularly (i.c.v.) administered salmon-calcitonin (S-CT), using the hot-plate test and the writhing test. The influence of the route of administration on the analgesia induced by morphine was also studied. After i.p. administration the analgesic effect was observed only in the writhing test. When S-CT was administered i.c.v., analgesia was observed in both tests, although it was greater in the writhing test than in the hot-plate test. I.c.v. injected S-CT increased the analgesia of i.p. injected morphine. Our results provide new information about the analgesic effect of S-CT and suggest that central mechanisms are involved.


Action of externally applied ATP on rat reticulospinal vasomotor neurons. In anesthetized rats, iontophoretic application of ATP excited the spinal cord-projection neurons in the rostral ventrolateral reticular nucleus of the medulla oblongata. The neuronal response to ATP was mimicked and then blocked by alpha,beta-methylene-ATP, a metabolically stable ATP analogue, and the response was abolished by suramin. Microinjections of ATP (3-100 pmol) into the rostral ventrolateral medulla produced a powerful pressor response. The results suggest that ATP may function as a neurotransmitter or neuromodulator involved in medullary regulation of cardiovascular functions.


D-cycloserine reverses the working memory impairment of hippocampal-lesioned rats in a spatial learning task. It is shown that D-cycloserine has cognition-enhancing properties in quinolinic acid hippocampal-lesioned rats. Lesioned rats had a severe impairment of working memory in an allocentric spatial reversal paradigm in the 8-arm maze, and performance could be restored with 12 mg/kg i.p. D-cycloserine, given 30 min before testing. The present findings favour the testing of D-cycloserine for clinical efficacy in patients with Alzheimer's disease with loss of pyramidal neurons and disconnected glycine-NMDA receptor activation.


Ethanol can inhibit nitric oxide production. Endogenous nitric oxide (NO) in exhaled air from anaesthetized rabbits was monitored by chemiluminescence. Ethanol (3-30 mmol kg-1) infused i.v. dose dependently reduced the levels of exhaled NO, with an IC50 of 23 +/- 3 mmol kg-1. L-Arginine (1 g kg-1) did not reverse the effect of ethanol. These results demonstrate the inhibition of NO formation by ethanol in vivo.


Postnatal behavioral development in methylazoxymethanol-induced microcephalic rats--a behavioral teratology study. Behavioral testings in methylazoxymethanol (MAM)-induced microcephalic rats were conducted. Pregnant Sprague-Dawley rats were treated intraperitoneally with 0, 20 or 40 mg/kg of MAM once a day on day 14 of gestation and were allowed to delivery. Male pups from each litter were examined for open field test at 6 weeks of age and shuttle-box avoidance test at 7 weeks or more of age. In the open field activity of pups, the counts of ambulation and locomoting distance in 40 mg/kg group have increased significantly as compared with those in control group. In the shuttle-box avoidance test, the avoidance response rate was dose-dependently high in the session of the 1st day. As to the interaction between the avoidance response rate and sequence of sessions, however, the avoidance response rate in 40 mg/kg group was significantly low. Rate of the rats with errors and number of response during the intertrial interval was significantly high in 40 mg/kg group. Thus, we could demonstrate functional disturbance in the memory retaining ability in utero MAM-exposed rats.


[Biological effects of exposure to high frequency electromagnetics on rabbits and guinea pigs]. To investigate the biological effects of exposure to feeble high frequency electromagnetism, skin surface temperature, blood vessel (arterioles and venules) diameter were examined, using infrared thermography, a laser doppler flowmeter, and a video microscope, respectively, in the ear of rabbits. After exposing the ear of rabbits to high frequency electromagnetism value of 9 MHz for 15 minutes, continued rising of local temperature was demonstrated. Though dilatation of arterioles was not seen. In addition, venules tended to dilate and blood flow also to increase, and microcirculation was accelerated at the site where electromagnetism was exposed. Hazardous effects of long term exposures of high frequency electromagnetism (9 MHz for 30 days, 8 hours/day) on guinea pigs were not observed in their behavior, food consumption, body and organ weights, hematological and biochemical values, macroscopic and microscopic findings on autopsy.


[Morphometric and immunohistochemical studies of the suprachiasmatic nucleus in the hereditary microphthalmic rat]. Morphometric and immunohistochemical analyses of the suprachiasmatic nucleus (SCN) were performed on hereditary microphthalmic rats. In normal rats, the number of cells and the volume of the SCN were 11, 631 and 6.7 x 10(-2) mm3 (an average taken from 12 SCNs). However, the neuronal population and volume of the SCN in hereditary microphthalmic rats were 7,450 and 4.5 x 10(-2) mm3 (an average taken from 14 SCNs), respectively. There were no significant differences in the size of neurons between normal and microphthalmic SCN neurons. Immunohistochemical studies showed that a considerable number of antivasopressin positive neurons were present in microphthalmic rats, despite their lack of the optic nerve. However, further detailed studies revealed that the number of antivasopressin positive neurons present in microphthalmic rats was only 68% of those found in normal rats. These findings suggest that the complete development of the SCN and vasopressin neurons depends on the visual input.


[Cryopreservation of unfertilized rat oocytes by ultrarapid freezing]. Unfertilized rat oocytes were placed in a highly concentrated solution of cryoprotectant (DAP 224:2M dimethylsulphoxide, 2M acetamide, 4M propylene glycol in PB1) in 0.5 ml sampling tubes and then immediately immersed into liquid nitrogen; thawing was conducted in a 37 degrees C waterbath. After thawing, 630 out of 968 oocytes (65.1%) were morphologically normal. After insemination in vitro of cryopreserved oocytes, the proportion of pronuclear oocytes with spermatail (s), male (s) and female pronuclei (8-10 h post insemination), and 2-cell embryos with two identical blastomeres (28-30 h post insemination) was 60.8% (152/250) and 29.8% (39/131), respectively. One hundred and fifty oocytes that were judged as pronuclear oocytes under the inverted microscope 8-10 h after insemination were transferred to the oviducts of pseudopregnant recipients; 18.7% (28/150) of the oocytes developed to normal young.


The domestication of Crocidura dsinezumi as a new laboratory animal. The dsinezumi shrew (Crocidura dsinezumi), a small insectivore, has been bred for the first time as a laboratory animal. The original animals were captured using Sherman's live traps and transferred into wooden cages. After several generations they were housed in plastic cages. Their diet consisted of trout pellets, cat food, and water provided ad libitum. Monogamous pairs were housed together for 2-3 weeks for mating, and the male was separated from the female during delivery and nursing. In captivity, the reproductive activity was observed throughout the year and the gestation period was estimated at 28-30 days with a litter size of between 1 and 4 pups. Pups grew very rapidly, and reached adult body size (mean: male, 9.7 g; female, 8.3 g) and sexual maturation at 6-8 weeks of age. The reproductive life was estimated at one and a half years, while the longevity was approximately 2 years.


Spontaneous lesions in cynomolgus monkeys used in toxicity studies. Spontaneous lesions in wild-caught, laboratory-maintained cynomolgus monkeys used in drug-toxicity studies were examined histopathologically in an effort to better distinguish toxic changes from spontaneous lesions and assess the toxicity of drugs more exactly. In the liver and kidney, where toxic changes are observed frequently, many spontaneous lesions were observed. Infiltration of mononuclear cells, vacuolization of the hepatocytes, dilatation of the renal tubules, and vacuolization of the renal epithelia were observed at a relatively high frequency. It is considered important to examine these changes carefully, because they closely resemble the changes recognized as toxic. Deposition of brownish pigment was observed in various organs such as the liver, kidney, spleen, intestinal tract, lung and brain, however the type of pigment differed among the organs, and histochemical examination revealed anthracosis or accumulation of hemosiderin, or melanin. Since the monkeys were caught in the wild, many parasitic lesions were observed especially in the large intestine and liver. Helminthous worms were frequently observed in the granulomas in the large intestine, however, no parasites were observed in the granulomas in the liver. Such lesions in the liver may be misinterpreted as toxic changes, when only scars or inflammatory lesions are observed.


[Immunocytochemical and morphometric studies on the development of renin-immunoreactive cells in the rat kidney]. The development of renin-immunoreactive cells was immunocytochemically studied in Wistar-Imamichi rats. The renin-immunoreactivity was localized in the walls of the abdominal aorta, renal artery and arterioles in the kidney. The renin-immunoreactive cells converged with the progress of development from the renal artery via several arterioles to the afferent arteriole in the kidney. In the afferent arteriole, renin-immunoreactive cells appeared only in those of matured glomeruli. The matured glomerulus first appeared at 17 days of gestation followed by the initial appearance of renin-immunoreactive cells at 18 days. The juxtaglomerular index showing the relation between matured glomeruli and renin-immunoreactive cells increased rapidly until 20 days of gestation, shifted thereafter at a level similar to that at 20 days until 7 days after birth, and increased again until 20 days after birth. Although the area of the transverse section at the hilus of the kidney increased all through the period examined, the number of matured glomeruli increased rapidly from 17 days of gestation to 7 days after birth and maintained a fixed level thereafter. This led to the decrease in the number of matured glomeruli per unit area after 7 days after birth. The fine structure of renin-immunoreactive cells showed no difference according to their sites of localization.


[Effect of sex hormones on the onset of diabetic syndrome in WBN/Kob rats]. The effects of sex hormone on diabetic conditions were investigated in WBN/Kob strain rats, i.e., castrated or spayed, hormone-treated, and non-treated rats. The effects of sex hormone on glycosuria, body-weight change, glucose tolerance and histopathology of the pancreas were compared among these animals. There were no abnormal changes in these parameters in the non-treated females and estrogen-treated males. The glycosuria began to be observed from the age of about 30 weeks in the non-treated group and from the age of 52 weeks in the castrated group. In the female animals, this symptom began to appear from the age of 55 weeks in the testosterone-treated group and from the age of 72 weeks in the spayed group. Before the onset of the diabetic symptoms, glucose tolerance was impaired in these animals. Body weights of the castrated and estrogen-treated males were lower than that of the non-treated males, especially in the estrogen-treated males. Those of the spayed and testosterone-treated females were much heavier than that of the non-treated females. Testosterone treatment accelerated body-weight gain in the spayed female animals. Histopathological examination of the pancreas revealed atrophy of the aciner tissue and atrophy and disappearance of the islet cells similar to those of the non-treated WBN/Kob male rats in the castrated males, spayed females and testosterone-treated females. However, these changes were not observed in the non-treated females or estrogen-treated males. These findings suggest that female hormone suppressed the onset of hyperglycemia along with glycosuria and male hormone accelerates the onset of hyperglycemia in the WBN/Kob rats.


Fine structure of the parotid gland in the crest-tailed marsupial-rat (Dasyuroides byrnei). The parotid gland of Dasyuroides byrnei was examined by light microscopy, and transmission and scanning electron microscopy. The acini were composed predominantly of seromucous cells with a few mucous cells. The seromucous cells were light or dark cells containing acidophilic spherical granules of moderate to high electron density and had well-developed cytoplasmic organelles-ordinary mitochondria and large mitochondria with tubular cristae, RER with vesicular or tubular elements, and Golgi apparatus with lamellae, vesicles and vacuoles. The mucous cells had basophilic amorphous granules of low electron density, like those of ordinary mucous cells. The intercalated ducts were composed of simple cuboidal light cells having a few electron-dense granules. The striated ducts consisted of tall columnar light cells containing numerous vesicles and mitochondria with tubular cristae, the same as found in acinar seromucous cells.


[Hematological and serum biochemical effects of nursing on the mother in cynomolgus monkeys (Macaca fascicularis)]. The effects of nursing on maternal hematological and serum biochemical values were analyzed in cynomolgus monkeys reared in indoor cages at Tsukuba Primate Center. In our breeding system, infants are usually separated from their mothers at the age of 121 to 180 days. Mother monkeys of such infants were studied hematologically and biochemically (Group B), as were mother monkeys who happened to have nursed their infants past 181 days after parturition (Group A). During the period with their infants, mother monkeys in the latter group showed lower white blood cell counts (WBC) and higher red blood cell counts (RBC), hematocrit values (Ht) and blood urea nitrogen concentrations (BUN) than the mother monkeys who had been separated from their infants. Also, serum calcium concentrations (Ca) were decreased with prolonged nursing periods, indicating that lactation by the mother monkey probably continues even for a period from 181 days to about one year after parturition if she nurses her infant. Lactation during this period may accelerate hematogenesis and protein metabolism in the mother monkey.


[Diurnal rhythm of drinking activity in the house musk shrew]. A diurnal rhythm of drinking activity in 7 male and 6 female house musk shrews (Jic: SUN) aged about one year was observed over a period of 10 days under a schedule of 12 hr light and 12 hr darkness (light on at 07:00). In general, the pattern of drinking activity was similar among both sexes, with around 24-hr diurnal rhythm. A few typical drinking patterns of these animals were represented as follows: 1) Drinking interval was very close in the dark phase, while it was a little too sparse in the light phase (n = 4). 2) Its interval remains stationary through a whole day (n = 5). 3) Drinking was performed between the latter half of light and the first half of dark phases (n = 4).


Production of normal young following insemination of frozen-thawed mouse spermatozoa into fallopian tubes of pseudopregnant females. Mouse epididymal spermatozoa in the cryopreservation solution (18% raffinose and 3% skim milk in distilled water) were frozen and stored at -196 degrees C, and later thawed at room temperature. The thawed sperm suspension was inseminated into the Fallopian tubes containing ovulated oocytes in pseudopregnant females on the day of finding the vaginal plug. Five out of 12 females gave birth to 28 Young (5.6 per liter).


Behavioral study on the gracile axonal dystrophy (GAD) mutant mouse. We investigated motor function and pain sensation in the gracile axonal dystrophy (GAD) mutant mouse, using the tail-flick test and the rotarod test. GAD (gad/gad) and normal sib mice (gad/+ or +/+) were used between 5 and 11 weeks of age, during which time the behavioral signs of GAD mice shifted from sensory ataxia (about 4 to 8 weeks of age) to paresis (after about 9 weeks of age). In the tail-flick test, significant shortening of latency was observed at 6 and 8 weeks of age in female GAD mice, in comparison with normal female mice. This may be related to dysfunction or degeneration of axons in the fasiculus gracilis, whose collaterals are thought to control the transmission of nociceptive information. In the rotarod test, a cumulative chi 2 test showed significant reduction in the performance times of GAD mice beginning at 5 and 6 weeks of age in males and females, respectively, indicating that the rotarod test can detect the development of motor incoordination as early as these ages. The performance times of GAD mice dropped sharply from 9 weeks of age onwards, and this is believed to reflect the progression of paresis. The rotarod test therefore appears to be a good method of quantifying behavioral changes in GAD mice and to be applicable both to objective selection of GAD mice before 8 weeks of age and to evaluation of drugs to treat ataxia or paresis.


Lymphocyte blastogenesis in hairless dogs following stimulation by various mitogens. Lymphocyte blastogenesis in hairless descendants of Mexican hairless dogs was examined using the following mitogens: phytohemagglutinin (PHA-M), Concanavalin A (Con A), lipopolysaccharide (LPS) and pokeweed mitogen (PWM). Blastogenetic responses to these mitogens were measured by glucose consumption test (GCT) and compared with those of haired beagle dogs. The response to PHA-M was significantly less in the hairless dogs than in the beagles. No significant differences in the responses to Con A, PWM and LPS were recognized. These results indicated T-cell dysfunction in the hairless dogs, coinciding with our previous work showing reduction of the delayed-type hypersensitivity reaction and early degeneration of the thymus.


[The influence of serum separators on biochemical values in experimental animals]. The influence of three serum separators, A, B and C, on biochemical values was examined. With A or B, changes in the values of ChE, LAP, UN, K, P, Cl and Ca in rat serum; ChE, UN, Cl and Ca in dog serum; and K and P in monkey serum took place over a period of 20 min after blood collection. Therefore, the biochemical values of whole blood were considered to be stable after 20 min. Thus, biochemical tests were conducted on serum from the three serum separators after allowing the blood to stand 20 min. The values obtained for each separator were not markedly different from those of the control. These results suggest that a serum separator is useful for separation of serum of experimental animals under the proper conditions.


[Cryopreservation of spermatozoa of a transgenic mouse]. Spermatozoa of a homozygous transgenic mouse, in which the firefly luciferase gene was expressed under the control of beta-actin promoter, were frozen at -196 degrees C. One fourth of the frozen sperm was later thawed and used for in vitro fertilization. Thirty-six of 65 oocytes (55.4%) developed to the 2-cell stage. All the 2-cell embryos were transferred to the oviducts of pseudopregnant recipients and 23 young (63.9%, 23/36) were born. All of young analyzed carried the transgene and showed the luciferase gene expression.


[Genetic profile of LIBP/1 inbred strain derived from the Kunming outbred stock of the mouse]. To make the genetic profile of the LIBP/1 inbred strain obtained from Kunming mice, the most widely used outbred stock in China, 26 loci were examined. The genotypes of four kinds of coat color genes were a/a, B/B, c/c and D/D. The results of testing 21 biochemical marker genes showed Akp-1b, Amy-1a, Car-2a, Ce-2a, Es-1b, Es-3a, Es-10a, Es-11a, Gpd-1a, Gpi-1a, Gus-1b, Hbbs, Idh-1a, Ldr-1a, Mod-1a, Mup-1b, Pep-3b, Pgm-a, Sep-1b, Tam-1c, and Trfb. The H-2 gene loci were Kb and Db.


[Developmental study of the ventral scent gland of the Mongolian gerbil]. Male and female in bred Mongolian gerbils aged 4, 5, 10, and 20 weeks were examined for the presence of a ventral scent gland macroscopically and histologically. It was found in about half of the gerbils aged 4 weeks and in all of the gerbils aged over 5 weeks. In adult male gerbils it weighed three times as much as in females. The ventral scent gland exhibited a sebaceous-like structure which consists of giant glandular cells with small vacuoles in the cytoplasm and the glandular cells displayed eosinophilic bodies contained within a duct, which are extruded through the lumen as holocrine-type secretion.


Navigating treatment impasses at the disclosure of incest: combining ideas from feminism and social constructionism. This article describes an approach to the social and emotional schisms that characterize the disclosure of intrafamilial sexual abuse (incest). It argues that ideas from social constructionism and feminism can be combined in such a way that what appear as either/or choices become both--and possibilities. These include: social control versus therapy, shame versus pride, attachment to one's abusive partner versus attachment to one's injured child, and "justice" versus "care."


Transformations: a blueprint for narrative changes in therapy. When problematic-symptomatic behaviors are conceived as embedded, retained, and maintained in collective stories, therapy can be described as the transformative process by which patients, families, and therapists co-generate qualitative changes in those stories. An emphasis on narratives allows one to specify further how those transformations unfold at the more "micro" level of the exchanges that take place throughout the consultation. To that specification is devoted the core of this essay, which closes with a discussion of the clinical, training, and, especially, research potentials of this systematization.


The California Family Health Project: I. Introduction and a description of adult health. Little research has addressed patterns of family and health relationships that reflect both the scope and complexity of family life and the breadth and diversity of health. In the first of a series of articles, we describe the California Family Health Project, a study in which four large "domains" of family life (Structure/Organization, World View, Problem Solving, and Emotional Management) were mapped, described, and compared with a large battery of adult health measures. We first present a brief critical overview of the literature on family and health research, then explain our rationale, define our approach to the multivariate analysis of family and health data, and describe our sample of 225 community-based families. To prepare for analyses with the family variables, we next present descriptive data based on separate principal components analysis (PCA) and multidimensional scaling analysis (MDS) of 14 self-reported health scores for husbands and for wives. No grouping or clustering of health variables emerged for either husbands or wives in the PCAs. A two-dimensional MDS analysis for husbands and for wives displayed the health variables in a circular pattern in which no predominant descriptive dimension or group of discrete dimensions emerged. Consequently, we decided that all 14 health scores will be used in the analyses, with the family variables to follow.


The California Family Health Project: II. Family world view and adult health. This article explores the pattern of relationships between family World View and adult Health in a community-based sample of 225 families. Family World View refers to the beliefs, appraisals, and values that define a family's orientation to the world. The interrelationships among eight self-reported family World View variables are described, using principal components analyses (PCA) and multidimensional scaling analyses (MDS). Derived, joint-spouse World Views also are examined using inter-battery factor analysis. The World View variables then are analyzed as a set with 14 self-reported health variables for husbands and wives separately, using canonical correlation. The PCAs for family World View yielded poor solutions for both husbands and wives. The MDS displayed the eight variables in a circular pattern for husbands and for wives, indicating the absence of a single broad dimension, or subgroupings of separate dimensions, that could be used to "describe" the domain. In the canonical analyses, family World View was a strong correlate of Health, with approximately 50% of the variance accounted for by the respective canonical variates. For husbands, what we called Family Coherence, Family Religiousness, Family Life Engagement, and Family Optimism, were correlated with Health. For wives, Family Coherence, Family Religiousness, and Family Optimism, were correlated with Health. Different patterns of health scores emerged by gender, with behavioral indicators, such as Smoking and Drinking, more salient for husbands, and mood indicators, such as Anxiety and Depression, more salient for wives.


The California Family Health Project: III. Family emotion management and adult health. This article explores the broad patterning of interrelationships between family Emotion Management and adult Health in a community-based sample of 225 families. Emotion Management refers to how emotion is expressed, acknowledged, and managed by the marital partners. Fifteen reliable-observer ratings of husband-wife behavior were made during each of three 10-minute Emotion Management Interaction Tasks (EMITs). Each of the three tasks "pulled" for the expression of a different emotional theme: loss, intimacy, or conflict. A principle components analysis of the 15 ratings yielded a poor solution. A nonmetric, multidimensional scaling analysis described a two-dimensional, bipolar display with ordering among the variables in each of two wings. One dimension reflected positive versus negative Emotion Management characteristics, and the second reflected active and overt versus passive or covert emotional expression. The pattern of relationships among the couple ratings was similar for each of the three tasks. Using canonical correlation, the couple ratings demonstrated significant associations with 14 adult Health scores for both husbands and wives for the intimacy and conflict tasks, but not for the loss task. Specific gender patterns also emerged. In general, couple overt emotional aversiveness was negatively associated with husbands' health, and couple emotional avoidance/distance was negatively associated with wives' health.


From front line to home front: a study of secondary traumatization. While the long-term effects of combat trauma on veterans have been studied extensively, its impact on veterans' wives has yet to be investigated. This study examined the implications of combat-induced psychopathology--wartime combat stress reaction (CSR) and current posttraumatic stress disorder (PTSD)--in a sample of 205 wives of Israeli combat veterans of the 1982 Lebanon war. Results show that both CSR and PTSD were associated with increased psychiatric symptoms in the wives. In addition, current PTSD was particularly found to contribute to impaired social relations among veterans' wives in a broad range of contexts, from inner feelings of loneliness, through impaired marital and family relations, and extending to the wider social network. Implications of these findings for treatment and further research are discussed.


Behavioral problems in sons of incarcerated or otherwise absent fathers: the issue of separation. The decade of the 1980s witnessed more than a doubling in the number of incarcerated individuals. Little is known about the psychological reactions of children whose parents are incarcerated, although a variety of behavioral disorders apparently related to separation, stigma, and deception of the child has been reported. The possibility of aggressive or antisocial behavior emerging in sons of incarcerated fathers has been mentioned as of particular concern in some reports. This article discusses salient themes in the literature on the reactions of children to parental incarceration, with an emphasis on boys' reactions to incarceration of their fathers. It critiques this literature and compares the findings with literature on the effects of separation in father absence related to other causes (for example, divorce, death, military service). Behavioral or emotional disorder associated with paternal incarceration probably is related mainly to associated factors such as the meaning of the incarceration to the child, the remaining caretaker's psychological characteristics and psychopathology, the parenting relationship between the caretaker and the child, and the coping capacities and resources of the family, rather than to the separation itself. Recommendations for further research in this increasingly important field are provided.


Purification, inhibitory properties and amino acid sequence of a new serine proteinase inhibitor from white mustard (Sinapis alba L.) seed. A new serine proteinase inhibitor, mustard trypsin inhibitor 2 (MTI-2), has been isolated from white mustard (Sinapis alba L.) seed by affinity chromatography and reverse phase HPLC. The protein inhibits the catalytic activity of bovine beta-trypsin and bovine alpha-chymotrypsin, with dissociation constants (Kd) of 1.6 x 10(-10) M and 5.0 x 10(-7) M, respectively, at pH 8.0 and 21 degrees C, the stoichiometry of both proteinase-inhibitor complexes being 1:1. The amino acid sequence of MTI-2, which was determined following S-pyridylethylation, is comprised of 63 residues, corresponding to a molecular weight of about 7 kDa, and shows only extremely limited homology to other serine proteinase inhibitors.


Effects of okadaic acid on the activities of two distinct phosphatidate phosphohydrolases in rat hepatocytes. Incubation of hepatocytes with okadaic acid displaced the N-ethylmaleimide-sensitive phosphatidate phosphohydrolase from the membrane fraction into the cytosol and partially prevented the oleate-induced movement of phosphohydrolase from cytosol to membranes. However, higher concentrations of oleate still caused translocation and activation of the phosphohydrolase. This enzyme is stimulated by Mg2+, and is probably involved in glycerolipid synthesis. Okadaic acid also decreased the concentration of diacylglycerol within the hepatocytes. Okadiac acid had no observable effect on the activity of an N-ethylmaleimide-insensitive phosphatidate phosphohydrolase which remained firmly attached to membranes. This activity is not stimulated by Mg2+ and is probably involved in signal transduction by the phospholipase D pathway.


Phorbol ester-induced secretion of human hepatocyte growth factor by human skin fibroblasts and its inhibition by dexamethasone. Human skin fibroblasts secreted a certain amount of human hepatocyte growth factor (hHGF), as determined by an enzyme-linked immunosorbent assay for hHGF. This hHGF secretion was remarkably stimulated by protein kinase C (PKC)-activating phorbol esters, which was inhibited by the simultaneous addition of dexamethasone. Pretreatment with phorbol 12-myristate 13-acetate (PMA) caused a down-regulation in hHGF secretion. hHGF secreted by the PMA-treated cells showed a potent hepatocyte growth-promoting activity which was neutralized by an anti-hHGF antiserum. These results indicate both that PMA-treated human skin fibroblasts produce biologically active hHGF and the possible involvement of PKC activation in this process.


The consensus sequences for cdc2 kinase and for casein kinase-2 are mutually incompatible. A study with peptides derived from the beta-subunit of casein kinase-2. Two series of synthetic peptides that reproduce the amino- and carboxyl-terminal segments of the beta-subunit of casein kinase-2, including the sites phosphorylated by CK2 and cdc2 kinase, respectively, have been used as model substrates for these enzymes. The N-terminal peptide beta(1-9), MSSSEEVSW, is readily phosphorylated by CK2 but not all by cdc2. The opposite is true of the C-terminal peptide beta(206-215), NFKSPVKTIR, whose Ser-4 is a good target for cdc2 while being unaffected by CK2. The individual substitutions of Pro-5 and Lys-7 in the latter peptide with Gly and Ala (or Glu), respectively, prevent its phosphorylation by cdc2, whereas the substitution of Lys-3 with Ala is well tolerated and the substitution of the target Ser with Thr actually improves phosphorylation. Thus the consensus sequence for cdc2 is shown to be X-S-P-X-K. Such a requirement for a basic residue at position +3 is opposite to that of CK2 whose consensus sequence (S-X-X-E/D/Yp/Sp) includes an acidic residue at the same position. Moreover the motif Ser-Pro is detrimental for CK2, preventing the phosphorylation of otherwise suitable peptides. These observations would rule out the possibility that the site specificity of CK2 might overlap with that of cdc2 and possibly of other Pro-directed protein kinases.


Unimpaired effect of insulin on glucokinase gene expression in hepatocytes challenged with amylin. Amylin appears to interfere with the action of insulin in muscle and possibly in liver. We have attempted to detect a direct antagonism between amylin and insulin in cultured rat hepatocytes. The stimulation of glucokinase gene expression was used as a marker of insulin action. Amylin proved ineffective in suppressing subsequent accumulation of glucokinase mRNA in response to maximal or submaximal doses of insulin. When applied to cells already induced by prior incubation with insulin alone, amylin failed to reverse induction, in contrast to the effectiveness of glucagon under the same conditions. Thus, amylin is not a physiological antagonist of insulin in the control of hepatic glucokinase gene expression.


Rapid kinetics of the interaction between daunomycin and drug-sensitive or drug-resistant P388 leukemia cells. The initial stages of the interaction of daunomycin (DNM) with drug-sensitive (P388/S) and drug-resistant (P388/100) cells have been defined by a rapid kinetics stopped-flow procedure. The process can be described by two kinetic components. The faster component accounts for rapid occupation of cell surface sites by DNM, as supported by experiments with liposomes with different surface charge. On the other hand, the effect of verapamil in the assays, suggests that the slower component is involved in the transport of the drug into the cells. Our observations are consistent with a loss in the control of the passive permeability to the drugs in the drug-resistant tumor cells.


A gene family homologous to the S-phase specific gene in higher plants is essential for cell proliferation in Saccharomyces cerevisiae. Previously we reported the isolation and characterization of the gene, cyc07, which was specifically expressed in the S phase during the cell cycle in synchronous cell division cultures of the higher plant, Catharanthus roseus. We found that the yeast Saccharomyces cerevisiae contains two closely related genes which show a high degree of similarity (about 64% at the amino acid level) to cyc07 of C. roseus. Site-directed disruption mutations demonstrated that the two yeast genes, homologous to cyc07, constitute an essential gene family for cell proliferation in yeast cells. Furthermore, the rate of cell proliferation varied with the gene copy number.


A dysfunctional C1 inhibitor protein with a new reactive center mutation (Arg-444-->Leu). A P1 mutation (Arg-444-->Leu) was identified in a dysfunctional C1 inhibitor from a patient with type 2 hereditary angioneurotic edema. The mutation was defined at the level of the protein (by sequence analysis of the Pseudomonas aeruginosa elastase-derived reactive center peptide), and the mRNA (CGC-->CTC) (by sequence analysis of PCR-amplified DNA).


Pine stilbene synthase cDNA, a tool for probing environmental stress. Stilbene synthase cDNAs were isolated from a pine (Pinus sylvestris) cDNA library. Poly(A)+RNA required for the preparation was obtained from young seedlings challenged with Botrytis cinerea. A full-length cDNA encoding pinosylvin-forming stilbene synthase was sequenced, and the deduced amino acid sequence was compared with sequences of resveratrol-forming stilbene synthases. The cDNA coding for the key enzyme of pinosylvin formation is a valuable tool for detecting early effects of environmental stress in pine needles.


Cloning and characterization of the rat glutathione peroxidase gene. The increased activity of glutathione peroxidase (GSHPx) in rat lungs is associated with the development of tolerance of the animals to hyperoxia. To understand further the regulation of expression of this enzyme, the molecular structure of the corresponding rat gene was characterized. The rat GSHPx gene consists of two exons interrupted by a single intron of 217 base pairs. The same initiation sites for transcription were found to be utilized in both lung and liver. The promoter of the GSHPx gene contains neither a 'TATA' box nor a 'CAAT' box. Instead, it comprises two copies of Sp1 binding motif and one copy of AP-2 binding motif. These features of the promoter may offer a clue to the mechanisms by which the expression of this gene is controlled.


Rainbow trout cytochrome P-450c17 (17 alpha-hydroxylase/17,20-lyase). cDNA cloning, enzymatic properties and temporal pattern of ovarian P-450c17 mRNA expression during oogenesis. A cDNA clone encoding cytochrome P-450c17 (17 alpha-hydroxylase/17,20-lyase) was isolated from a rainbow trout ovarian follicle cDNA library. The cDNA contained an open reading frame of 1,542 nucleotides encoding a protein of 514 amino acid residues. The amino acid sequence of trout P-450c17 shows a much greater homology with chicken P-450c17 than with that of human, bovine and rat. The trout P-450c17 expressed in non-steroidogenic mammalian COS-1 cells showed both 17 alpha-hydroxylase and 17,20-lyase activities. The cDNA only hybridized to a single species of mRNA (2.4 kb) isolated from rainbow trout ovaries; the 2.4 kb transcripts were abundant in trout ovaries during the later stages of oogenesis.


cDNA cloning of rat proteasome subunit RC1, a homologue of RING10 located in the human MHC class II region. The nucleotide sequence of a cDNA that encodes a new subunit, named RCl, of rat proteasomes (multicatalytic proteinase complexes) has been determined. The polypeptide predicted from the open reading frame consisted of 208 amino acid residues with a calculated molecular mass of 23, 130, which is consistent with the size obtained by electrophoretic analysis of purified RCl. The partial amino acid sequences of several fragments of RCl, obtained by protein chemical analyses, were found to be in excellent accordance with those deduced from the cDNA sequence. Surprisingly, the overall structure of RCl was found to be almost identical to that of recently isolated RING10, whose gene is located in the class II region of the human MHC gene cluster. This finding suggests that RCl is a homologue of human RING10, supporting the proposal that proteasomes are involved in the antigen processing pathway.


Development of insulin sensitivity in rat skeletal muscle. Studies of glucose transporter and insulin receptor mRNA levels. Expression of GLUT-4 and insulin receptor mRNAs was investigated in rat skeletal muscle by Northern hybridization. GLUT-4 mRNA was barely detectable in foetal muscle, was expressed at low levels by 1-8 days and at 2-3-fold higher levels during and after weaning (18-40 days). In contrast there was little change in insulin receptor mRNA levels prior to weaning and a reduction in mRNA abundance between 18 and 40 days. Weaning rats on to a diet rich in fat prevented the increase in GLUT-4 abundance seen between 15 and 29 days in animals weaned on a high-carbohydrate diet.


Expression of 9 Salmonella typhimurium enzymes for cobinamide synthesis. Identification of the 11-methyl and 20-methyl transferases of corrin biosynthesis. Nine of the cbi genes from the 17.5 kb cob operon of Salmonella typhimurium previously shown by genetic studies to be involved in the biosynthesis of cobinamide from precorrin-2, have been subcloned and expressed in Escherichia coli. Seven of the gene products were found in the soluble fraction of cell lysates and have been purified. The gene products corresponding to cbi E, F, H and L were shown by SAM binding and by homology with other SAM-binding proteins to be candidates for the methyltransferases of vitamin B12 biosynthesis. The enzymatic functions of the gene products of cbiL and cbiF are associated with C-methylation at C-20 of precorrin-2 and C-11 of precorrin-3.


A consensus repeat sequence from the human insulin gene linked polymorphic region adopts multiple quadriplex DNA structures in vitro. A hypervariable region consisting of repeats of a 14 base pair (bp) consensus sequence ACAGGGGT(G/C)(T/C)GGGG is located 363 bp upstream of the human insulin gene. Different repeat numbers of this oligonucleotide give rise to a polymorphism, and so this region is commonly known as the insulin gene linked polymorphic region (ILPR). Here we present evidence, based on the mobility in non-denaturing polyacrylamide gels of two dissimilarly sized oligonucleotides containing the ILPR consensus sequence, that this sequence can adopt a number of quadriplex DNA structures in vitro.


Cloning and sequence analysis of the phenylalanyl-tRNA synthetase genes (pheST) from Thermus thermophilus. While crystals suitable for X-ray diffraction analyses are available of phenylalanyl-tRNA synthetase (PheRS) from the thermophilic bacterium Thermus thermophilus, neither the primary structure of its constituent alpha and beta subunits nor the nucleotide sequence of the corresponding pheS and pheT genes were known. Using specific oligonucleotides of conserved pheS regions that were adapted to the T. thermophilus codon usage, we identified, cloned and subsequently sequenced the pheST genes of this bacterium. The sequences reported here will greatly aid in the three-dimensional structure determination of T. thermophilus PheRS, a heterotetrameric (alpha 2 beta 2), class II aminoacyl-tRNA synthetase.


Expansion of the mammalian 3 beta-hydroxysteroid dehydrogenase/plant dihydroflavonol reductase superfamily to include a bacterial cholesterol dehydrogenase, a bacterial UDP-galactose-4-epimerase, and open reading frames in vaccinia virus and fish lymphocystis disease virus. Mammalian 3 beta-hydroxysteroid dehydrogenase and plant dihydroflavonol reductases are descended from a common ancestor. Here we present evidence that Nocardia cholesterol dehydrogenase, E. coli UDP-galactose-4 epimerase, and open reading frames in vaccinia virus and fish lymphocystis disease virus are homologous to 3 beta-hydroxysteroid dehydrogenase and dihydroflavonol reductase. Analysis of a multiple alignment of these sequences indicates that viral ORFs are most closely related to the mammalian 3 beta-hydroxysteroid dehydrogenases. The ancestral protein of this superfamily is likely to be one that metabolized sugar nucleotides. The sequence similarity between 3 beta-hydroxysteroid dehydrogenase and the viral ORFs is sufficient to suggest that these ORFs have an activity that is similar to 3 beta-hydroxysteroid dehydrogenase or cholesterol dehydrogenase, although the putative substrates are not yet known.


Stimulation of glucose transport in L6 muscle cells by long-term intermittent stretch-relaxation. Skeletal muscle stretch increases resting metabolism and causes hypertrophy. We have examined the effect of mechanical stretch in vitro on glucose transport activity and transporter contents in L6 muscle cells. Long-term (24-48 h) stretch-relaxation (25% maximal elongation at 30 cycles per min) of cell monolayers significantly increased glucose uptake by 1.6- to 2-fold in myotubes but not in myoblasts. The presence of serum was required for the stretch-relaxation induced increase in glucose uptake. Cycloheximide inhibited the mechanical stimulation of glucose uptake, and the latter response was not additive to the stimulatory effect of long-term exposure to insulin. GLUT1 and GLUT4 glucose transporter contents were not changed in total cell membranes from mechanically stimulated cells relative to controls. These results indicate that mechanical stimulation through passive stretch may be an important regulation of nutrient uptake in fetal myotubes independent of innervation.


Glycation of crystallins in lenses from aging and diabetic individuals. Water-soluble crystallins were obtained from clear human lenses of different age (4-81-year-olds) and lenses of individuals showing senile or diabetic cataracts. Levels of early glycation products were high in the high molecular weight material (HM) and the alpha-crystallin fractions, compared with beta- and gamma-crystallins. This difference becomes more prominent upon aging. The content of total early glycation products in HM and alpha-crystallin increases clearly with age, whereas levels remain relatively constant in the beta- and gamma-crystallins. There is an elevation of early products in cataractous lenses from diabetic individuals compared with those suffering from senile cataract. Specific non-tryptophan fluorescence (excitation/emission wavelengths 370/440 nm), used as an indicator for late glycation products, increased dramatically with age and was 2-fold higher in the diabetic subjects. Levels of fluorescence decreased in the order HM > alpha- > beta- > gamma-crystallins. The results suggest an increase in glycation rate in alpha-crystallin as a result of aging and diabetes, while the rate of glycation of beta- and gamma-crystallins remains almost constant.


Structure of UvrABC excinuclease-UV-damaged DNA complexes studied by flow linear dichroism. DNA curved by UvrB and UvrC. The interaction between UvrABC excinuclease from Escherichia coli and ultraviolet light-(UV) damaged DNA was studied by flow linear dichroism. The dichroism signal from DNA was drastically decreased in intensity upon incubation with UvrA and UvrB or whole enzyme in the presence of effector ATP. The change was specific for UV-damaged DNA, and a concluded suppressed DNA orientation suggests the wrapping of DNA around the protein. The incubation with the UvrC subunit alone also somewhat reduces the signal, however, in this case the change was smaller and not specific for UV-damaged DNA. The structural modification of DNA, promoted by the (UvrA2-UvrB) complex, probably facilitates or stabilizes the interaction of the UvrC subunit with DNA for the excision.


Correlation of cofactor binding and the quaternary structure of pyruvate decarboxylase as revealed by 31P NMR spectroscopy. The pH dependence of the quaternary structure of pyruvate decarboxylase (EC 4.1.1.1) has recently been discovered [(1990) FEBS Lett. 266, 17-20; (1992) Biochemistry (in press)]. In the present study we have investigated the change in quaternary structure by observing the binding of the cofactor, thiamine pyrophosphate, using 31P NMR spectroscopy. The dissociation of the native tetramers into dimers when increasing the pH coincides with a weaker binding of the cofactor and loss of enzyme activity. The results provide further evidence that thiamine pyrophosphate is bound primarily via the beta-phosphate moiety. In addition, a phosphoserine has been discovered in two of the four subunits.


Initiator-promoter coupling of phospholipases D and A2 in platelets upon cholesterol incorporation. Since phospholipases exist within a membrane lipid environment, it is not unreasonable to assume that cholesterol capable of changing the lipid environment can effect the coupling relationship among the signal transducing components. Our previous study showed that the 'molecular switch' through which membrane cholesterol modulates cyclic nucleotide levels and Na+/H+ exchange within human platelets is phospholipase A2. We demonstrate here that membrane cholesterol initiates the activation of phosphatidyl choline phospholipase D and phosphatidic acid thus generated promotes the activation of its phospholipase A2 in the presence of extraplatelet calcium. More important, inhibition of phospholipase D by zinc blocks the activation of phosphatidic acid phospholipase A2 in platelets upon cholesterol incorporation. Our result led us to postulate that membrane cholesterol induced initiation promotion coupling of phospholipases D and A2 in human platelets may be responsible for the hypersensitized state of platelets in hypercholesterolemic patients.


Na(+)-dependent AIB transport by neuroblastoma cells. Na(+)-dependent amino isobutyric acid transport by two neuroblastoma cell lines with and without amplification of the oncogene N-myc is studied. Surprisingly, the contribution of system A is greater in the cell line showing no N-myc amplification. Preliminary data support a role for essential tyrosine and cysteine residues in the active center of the carriers, mainly in system A.


Human peripheral blood lymphocytes express D5 dopamine receptor gene and transcribe the two pseudogenes. Sequential reverse transcription and polymerase chain reaction (RT-PCR) of the mRNA were used to investigate the expression of dopamine receptors in human peripheral blood lymphocytes. The RT-PCR products contained three types of sequences, each corresponding to those of the D5 dopamine receptor gene and the two related pseudogenes. The lymphocyte cDNA library also contained the clones encoding parts of the three genes. Binding profiles of dopaminergic ligands to the lymphocytes were similar to those for the native neuronal membranes.


Interaction of alpha subunit of GTP-binding protein Go with a 20-kDa Triton-insoluble membrane protein in bovine brain. Heterotrimeric Go bound to the membranes of bovine brain, but Go alpha remained bound to the membranes even after activation with GTP gamma S. Furthermore, Go alpha bound to a Triton X-100-insoluble fraction of the membranes in a saturable manner. However, the 37-kDa Go alpha eliminated by trypsin at the amino-terminus could not bind to the fraction. Using a blot overlay approach of the Triton-insoluble fraction, only a 20-kDa protein was identified that interacts with Go alpha. These results indicate that Go alpha binds to a 20-kDa Triton-insoluble protein in the bovine brain membranes.


The influence of oligodeoxyribonucleotide phosphorothioate pyrimidine strands on triplex formation. Analogues of the homopyrimidine oligonucleotide dT15 that contained phosphorothioate bonds of a mixture of diastereoisomers or one of the two stereoisomers (either Rp or Sp) were synthesized. The analogues were mixed under conditions conducive to the formation of triple-stranded assemblies. The mixtures were characterized by their thermal stabilities (Tm values), CD spectra, and gel electrophoresis pattern. The 34-mer duplexes containing 15 central purines on one strand and 15 complementary pyrimidines on the other strand gave no detectable triple helix upon combination with dT15S14. On the other hand, 34-bp duplexes with dT15S1, having Rp or Sp, formed triple helixes. This suggests that a steric factor plays an important role in triple helix formation.


A GC box in the bidirectional promoter is essential for expression of the human dihydrofolate reductase and mismatch repair protein 1 genes. The human dihydrofolate reductase and mismatch repair protein 1 genes are organized in a head-to-head configuration separated by an 88 base-pair segment and directed by a bidirectional promoter. In vivo transient assays of the site directed mutant promoters using firefly luciferase as a reporter showed that an AT-rich sequence, ACAAATA, in the GC-rich promoter sequence is not required for transcription. However, two out of four GC boxes were shown to function as bidirectional positive regulatory elements. Among them, a GC box at the midpoint of the region between the two initiation sites is essential for supporting minimal bidirectional activity.


Comparison of the Ca2+ binding properties of the gamma-carboxyglutamic acid-containing module of protein Z in the intact protein and in N-terminal fragments. Protein Z is a vitamin K-dependent plasma protein of unknown function. Its modular structure is identical with those of factors VII, IX, X, and protein C. These proteins have an N-terminal gamma-carboxyglutamic acid (Gla)-containing module which binds six to ten Ca2+. In factors IX, X, and protein C, the adjacent epidermal growth factor (EGF)-like module binds one Ca2+ whereas the EGF-like module in protein Z does not. We have compared the Ca2+ binding properties of a fragment of protein Z comprising the Gla and N-terminal EGF-like modules (pZ-GlaEGFN) with those of intact protein Z and the isolated Gla module by measuring the Ca(2+)-induced quenching of the intrinsic protein fluorescence. The similar Ca2+ affinities of pZ-GlaEGFN and protein Z indicate that pZ-GlaEGFN has a native conformation and normal Ca2+ binding properties. A comparison of the Ca2+ binding to pZ-GlaEGFN with those to the corresponding fragments of factors IX, X, and protein C indicate that Ca2+ binding to the N-terminal EGF-like modules in the latter proteins does not influence the folding and Ca2+ binding properties of their Gla modules. Furthermore, the Ca(2+)-induced fluorescence enhancements of GlaEGF fragments from factors IX, X, and protein C appear to be caused by Ca2+ binding to the site in the EGF-like modules since it is not observed for pZ-GlaEGFN.


Characterization of a spinach psbS cDNA encoding the 22 kDa protein of photosystem II. An intrinsic 22 kDa polypeptide is found associated with the oxygen-evolving photosystem II (PSII) core complex in all green plants and cyanobacteria so far examined, although it does not appear to be required for oxygen evolution. Amino acid sequence information obtained from the purified 22 kDa protein was used to construct a probe that was employed to isolate a full-length cDNA clone encoding the 274-residue precursor of the 22 kDa protein. Hydropathy plot analysis predicts the existence of four membrane-spanning helices in the mature protein. The two halves of the approximately 200-residue mature protein show high sequence similarity to each other, suggesting that the psbS gene arose from an internal gene duplication. The 22 kDa protein has some sequence similarity to chlorophyll a/b-binding proteins.


Complex formation between glutamyl-tRNA synthetase and glutamyl-tRNA reductase during the tRNA-dependent synthesis of 5-aminolevulinic acid in Chlamydomonas reinhardtii. The formation of a stable complex between glutamyl-tRNA synthetase and the first enzyme of chlorophyll biosynthesis glutamyl-tRNA reductase was investigated in the green alga Chlamydomonas reinhardtii. Apparently homogenous enzymes, purified after previously established purification protocols were incubated in various combinations with ATP, glutamate, tRNA(Glu) and NADPH and formed complexes were isolated via glycerol gradient centrifugation. Stable complexes were detected only after the preincubation of glutamyl-tRNA synthetase, glutamyl-tRNA reductase with either glutamyl-tRNA or free tRNA(Glu), ATP and glutamate, indicating the obligatory requirement of aminoacylated tRNA(Glu) for complex formation. The further addition of NADPH resulting in the reduction of the tRNA-bound glutamate to glutamate 1-semialdehyde led to the dissociation of the complex. Once complexed to the two enzymes tRNA(Glu) was found to be partially protected from ribonuclease digestion. Escherichia coli, Bacillus subtilis and Synechocystis 6803 tRNA(Glu) were efficiently incorporated into the protein-RNA complex. The detected complexes provide the chloroplast with a potential channeling mechanism for Glu-tRNA(Glu) into chlorophyll synthesis in order to compete with the chloroplastic protein synthesis machinery.


Natural human tumor necrosis factor beta (lymphotoxin). Variable O-glycosylation at Thr7, proteolytic processing, and allelic variation. Natural human tumor necrosis factor beta (TNF-beta) purified from supernatants of a human B-lymphoblastoid cell line was found to be heterogeneous in molecular mass, with seven components resolved by gel electrophoresis. All components are N-glycosylated at Asn62; N-glycosylation does not contribute to heterogeneity. In addition, part of the molecules are O-glycosylated at Thr7; O-glycosylation is heterogeneous due to variable decoration with neuraminic acid. The four lower molecular mass forms are derived from the full-length protein by trypsin-like proteolytic cleavage in the N-proximal region; these clipped molecules lack O-linked carbohydrates. Two allelic variants differing in amino acid position 26 (threonine/asparagine) were identified.


'All-or-none' mechanism of the molten globule unfolding. The Gdm-HCl-induced unfolding of bovine carbonic anhydrase B and S. aureus beta-lactamase was studied at 4 degrees C by a variety of methods. With the use of FPLC it has been shown that within the transition from the molten globule to the unfolded state the distribution function of molecular dimensions is bimodal. This means that equilibrium intermediates between the molten globule and the unfolded states are absent, i.e. the molten globule unfolding follows the 'all-or-none' mechanism.


Vessel repair after balloon angioplasty: morphological appearance and prostacyclin synthesising capacity. Immediately after balloon dilatation of the rabbit aorta the release of prostacyclin is diminished. In this study the morphological appearance and time course for recovery of prostacyclin production after balloon dilatation have been investigated. Healthy rabbit aortas were analysed 1 h (n = 12), 1 week (n = 13) and 1 month (n = 13) after angioplasty. The production of prostacyclin, from dilated and non-dilated aortic segments, was recorded in a perfusion system. Prostacyclin was measured as its stable degradation product 6-keto-PGF1 alpha. Scanning electron microscopy and light microscopy were used to analyse the type of cells present at the luminal surfaces of the segments. When endothelial cells were found their degree of coverage was also estimated. One hour after balloon dilatation there was a lower production of prostacyclin from the angioplasty segments than from controls. Also, the response to added arachidonic acid (AA) was lower in the angioplasty segments. No endothelial cells were present in the angioplasty segments. After 1 week there was no difference in the basic production of prostacyclin but there was still a lower response from angioplasty segments to the addition of AA. The inner surfaces of the angioplasty segments were covered by three to five layers of smooth muscle cells (SMC). After 1 month, there was no difference in either the basic production or after the addition of AA between control and angioplasty segments. The angioplasty segments were covered with a multilayer of SMC. The control segments had an almost complete cover of endothelial cells at every time interval after angioplasty.(ABSTRACT TRUNCATED AT 250 WORDS)


Management of acute arterial trauma of the upper extremity. During a 3.5 year period ending in September 1991, 50 patients with 51 arterial injuries of the upper extremity were operated on and followed up for more than 6 months. The majority of the cases suffered penetrating injuries, but in 18% blunt trauma led to thrombosis. The most frequently observed signs were haemorrhage, neurological disorders and a peripheral pulse deficit. The brachial artery was most frequently injured. Preoperative arteriography was performed in 20% and peroperative angiography in 16% of cases. Reconstructive surgery was performed in 88% and the brachial artery was ligated in a single patient who was in shock. An end to end anastomosis was possible in 55% and in 45% of cases an autogenous vein graft was used. A microsurgical technique was used in cases of forearm arterial injuries. In five patients with prolonged ischaemia, forearm fasciotomies were needed. Additional procedures such as venous, bone or nerve repair was also necessary. In four cases a temporary shunt was used. All patients were alive with late patency of 91% in the axillobrachial arterial segment. A single failure after repeated brachial artery reconstruction was observed following severe traumatic amputation with large tissue defects. Post-operative arteriography revealed a good anastomosis with patent grafts. Arterial reconstruction is clearly necessary in these injuries and we believe that an experienced vascular surgeon should be involved.


Amputation level is not adversely affected by previous femorodistal bypass surgery. Data was reviewed on patients presenting with lower limb-threatening ischaemia to a single vascular unit between September 1984 and December 1990. Three hundred and thirty patients underwent either femoro-popliteal or femoro-infrapopliteal reconstructive surgery and 316 primary amputations were performed on 281 patients. Sixty-three secondary amputations were performed following a femoro-distal bypass failure. The below-knee amputation to above-knee amputation ratio (BKA/AKA) was 1.0 in the primary amputation group to 0.91 in the secondary amputation group. A direct comparison cannot be made as the patients represent two separate clinical groups. The ratio of reconstructive operations to primary amputations more than doubled between the first and second halves of the study. The BKA:AKA ratio in the secondary amputation group and the total number of secondary amputations remained relatively constant. This study supports the view that previous femoro-distal surgery does not adversely affect the overall outcome of amputation.


The potential risk for subclavian vein occlusion in patients on haemodialysis. Subclavian vein (SCV) stenosis or occlusion can be a late complication of temporary haemodialysis or following catheterisation for intravenous hyperalimentation. In five patients with prior catheterisation or trauma of the SCV, incapacitating oedema of the upper extremity developed only after the creation of ipsilateral arteriovenous (AV) fistulas for haemodialysis. The duration of the previous catheterisation was 2 to 4 weeks. Massive upper extremity oedema developed at 10 days to 22 months with an average of 11 months after the establishment of AV fistulas. Subclavian-axillary vein bypass using a ringed polytetrafluoroethylene graft was successful for one patient and ligation of the AV fistulae led to good results for the other three. In planning vascular access procedures in the upper extremity, venography should be mandatory to try to prevent such complications if a previous history of subclavian catheterisation exits.


Femoropopliteal arterial reconstruction with intraoperative iliac transluminal angioplasty for disabling claudication: results of a combined approach. Seventeen patients with disabling claudication resulting from multilevel arteriosclerotic disease were treated by combined intraoperative iliac transluminal angioplasty and femoropopliteal arterial reconstruction. Clinical improvement or total relief of ischaemic symptoms was observed in 15 out of 17 patients. Iliac pressure gradients were reduced with balloon dilatation to < 2 mmHg in all cases. The mean (+/- S.D.) resting ankle-brachial systolic pressure index increased from 0.42 +/- 0.14 to 0.87 +/- 0.21. Complications from intraoperative angioplasty were not encountered and no early graft failures were seen. The primary actuarial graft patency at 1, 2 and 5 years was 100, 88 and 67%, respectively. Combined intraoperative iliac transluminal angioplasty and femoropopliteal arterial reconstruction is a useful alternative to conventional surgical revascularisation in the treatment of selected patients with disabling claudication in the presence of multilevel arteriosclerotic disease.


Infrarenal abdominal aortic aneurysms less than five centimetres in diameter: the surgeon's dilemma. A number of recent studies have brought new data and a better understanding of the risk versus benefit of abdominal aortic aneurysm repair. The present evidence indicates that the risk of rupture for non tender aneurysms less than 5 cm is so low that patients can generally be followed by 6 monthly ultrasound and not undergo immediate operation. Surgery should be the method of choice for most patients where the aneurysm is greater than 5 cm.


Peroperative anticoagulation with antithrombin or heparin in infrainguinal bypass surgery. Patients suffering from atherosclerosis may have a hypercoagulable state which is further aggravated by surgery. Thrombin, a central enzyme in the coagulation process, cleaves fibrinogen to fibrin. Therefore, inhibition of thrombin is an important anticoagulant mechanism. This is accomplished by heparin in concert with antithrombin III (AT), but vessel wall glycosaminoglycans may act as substitutes for heparin and catalyse thrombin inhibition. The present study examines whether administration of AT or heparin is effective as an anticoagulant during infrainguinal bypass surgery. Preoperatively and during surgery the patients had elevated levels of fibrinogen, fibrinopeptide A (FPA) and thrombin-antithrombin (T-AT) complexes. There were higher levels of FPA in the venous outflow from the ischemic leg than in the arterial inflow. Taken together these measurements indicate ongoing coagulation in the operated leg. Administration of heparin decreased FPA levels and prevented intraoperative graft thrombosis, whereas in patients receiving AT, T-AT levels increased but FPA levels were unchanged. In the latter group, intraoperative graft thrombosis occurred in a high proportion. Based on additional case histories in these patients with hypercoagulability, it is suggested that fibrinogen is a risk factor for thromboembolic complications and that a combination of low dose of heparin and AT might be an effective regimen to prevent intraoperative thrombosis with a low risk of haemorrhage.


Popliteal vein entrapment in the normal population. The aim of this study was to determine the incidence and severity of popliteal vein compression by full knee extension in the normal population. The popliteal veins in 100 healthy volunteers (200 limbs) with no history of previous deep vein thrombosis (DVT) or venous obstruction were examined using duplex scanning with the knee slightly flexed and then fully extended. Knee extension produced complete obstruction in 17 subjects and severe obstruction (< 50% decrease in diameter) in a further 10 subjects. Thirteen subjects had unilateral compression and 14 bilateral. The 27 subjects were tested for functional venous outflow obstruction with air plethysmography. In flexion, the outflow fraction was normal (> 40%) in all subjects. With the knee fully extended, severe or complete venous obstruction (outflow fraction < 10%) was found in eight subjects. Moderate obstruction (outflow fraction 10-40%) was found in all the remaining 19 subjects. When digital compression of the long saphenous vein was performed, these subjects also demonstrated severe outflow obstruction. Although the incidence of symptoms of functional venous obstruction is rare in the general population, these findings have important implications for venous stasis for patients on the operating table and in those having prolonged bed rest. Studies investigating the association between popliteal vein compression and postoperative deep venous thrombosis are needed.


Adverse outcome in surgery for chronic leg ischaemia--risk factors and risk prediction when using different statistical methods. to compare two different statistical methods in predicting the outcome of surgery for chronic leg ischaemia. the present study from the Swedvasc registry is based on an inception cohort of 1635 patients with chronic leg ischaemia (intermittent claudication in 609 and critical ischaemia in 1026 patients), who have been followed until 1 year after surgery. Outcome was classified as improved vs. not improved, amputated or dead in claudication and as the intact leg vs. amputation or death in critical ischaemia. logistic regression analysis was compared to the inductive expert system program, Assistant Professional, in the prediction of outcome. Seventy per cent of cases in the data base were used to create a risk factor model including 17 variables registered in Swedvasc. These variables included an assessment of patients overall health status, severity of disease, the surgeon's experience and surgical procedures. This model was then evaluated using the remaining 30% of the patients in the data base. a risk score indicating a probability of an adverse outcome exceeding 0.5 was, in patients with intermittent claudication, associated with a sensitivity of 38% using logistic regression and 26% using Assistant Professional. The percentages of correctly predicted adverse outcomes were 29 and 50%, respectively. In patients with critical ischaemia, the sensitivities with the two methods were 68 and 38% and the predictive values 42 and 57%, respectively. the risk scores created with the two methods gave low sensitivities. It is concluded that risk functions could not be used to predict an adverse outcome in patients operated on for chronic leg ischaemia with the data set used.


Growth factor release by smooth muscle cells is dependent on haemodynamic factors. Abnormal proliferation of smooth muscle cells (SMCs) is a key event in the development of the atherosclerotic plaque. The early atherosclerotic plaque localises preferentially in areas of flow separation. The basic mechanisms which correlate haemodynamic forces and atherosclerotic plaque formation are not known. Bovine arterial SMCs were subjected to increasing levels of shear stress for 24 h in an in vitro system. The control group was subjected to similar incubation conditions without flow. SMCs subjected to shear stress released a higher quantity of mitogens, including a platelet derived growth factor (PDGF)-like substance. This mitogenic activity was partially reduced (30%, p < 0.01) by an excess of monospecific anti-PDGF antibody. The release of mitogens was proportional to the level of shear stress and was still evident 24 h after flow cessation. In conclusion, shear stress influences the release of mitogens; this might represent a mechanism which links haemodynamic forces and atherosclerotic plaque formation.


Simultaneous resection of abdominal aortic aneurysms and early gastric cancer by retroperitoneal and transperitoneal approach. The surgical approach to patients with abdominal aortic aneurysm and gastrointestinal malignancy remains controversial. We experienced two cases with abdominal aortic aneurysm and gastric cancer, which were treated by a one-stage operation using a different approach. At first, the operation for the aneurysm was done through a retroperitoneal approach and then, a partial gastrectomy for gastric cancer was done through a transperitoneal approach. The postoperative course of both cases was uneventful. The patients were discharge on the 19th and 21st postoperative days, respectively. This one-stage operation using different isolated approaches, such as the retroperitoneal approach for abdominal aortic aneurysm and transperitoneal approach for gastric cancer, was useful for the patients with abdominal aortic aneurysm and particularly early gastric cancer in terms of preventing an infection of the prosthetic graft.


Co-variation between walking ability and circulatory alterations in patients with intermittent claudication. Unselected patients (n = 183) with subjective symptoms of intermittent claudication were examined clinically and by various circulatory tests (calf blood-flow, ankle, toe pressures). The aims of the present study were to evaluate to what extent the central or peripheral circulation is limiting in unselected patients with subjective symptoms of intermittent claudication, to determine the co-variation between the maximum walking capacity and traditional haemodynamical measures mentioned above and to evaluate to what extent a traditional bicycle ergometer exercise test and treadmill walking test give similar information regarding maximum performance. Eighty-five per cent of all patients were or had been smokers and 16% were diabetics. The mean ankle/brachial blood pressure index was 0.58 +/- 0.02 and the average post-ischemic maximum calf bloodflow was 13.3 +/- 0.6 ml/min/100 ml tissue. Leg arterial insufficiency was the limiting factor of walking capacity in 90% of all patients at 87 +/- 2 W corresponding to a walking distance of 282 +/- 13 m, while leg exhaustion was the limiting factor in 80% of the patients during test on the bicycle ergometer at maximum 84 +/- 2W. The mean maximum walking capacity for all patients was 86 +/- 3W and the mean maximum capacity on the bicycle ergometer was 87 +/- 2W. The ankle/brachial index showed only a weak correlation (r = 0.30, p < 0.002) to walking capacity. Our results demonstrate that the maximum walking capacity on a treadmill agrees with mean values of maximum exercise capacity on a bicycle ergometer.(ABSTRACT TRUNCATED AT 250 WORDS)


Epidemiology of abdominal aortic aneurysm: the effect of differing definitions. There have been several published definitions for abdominal aortic aneurysm (AAA), however, it is not known how different definitions affect what is reported. This paper presents an evaluation of three definitions taken from the literature on (a) aneurysm frequency, (b) prevalence rate and (c) prevalence ratio from an ongoing case-control study evaluating whether male siblings of patients with known AAA have a higher prevalence of aneurysms compared to a control group. Depending on the definition used, the frequency of AAA could vary from a low of five to a high of 23; the prevalence of AAA in siblings could range from 2.1 to 18.8. Changing definitions alters the prevalence ratio (risk of disease) in both magnitude and direction. On the one hand, male sibling of affected patients are at approximately one and a half times (1.6) greater risk of having an AAA compared to controls, whereas using a different definition they are at less than half the risk (0.3).


Endarterectomy of the superficial femoral artery: a procedure worth reconsidering. The preferred management of superficial femoral artery occlusive disease remains unresolved. The oldest technique for restoring vascular continuity, endarterectomy, has been largely replaced by bypass operations and percutaneous transluminal angioplasty. We have continued to perform semi-closed endarterectomy in selected cases and review here a series of 231 consecutive cases in 197 patients treated during the last 10 years. The indication for the endarterectomy was disabling claudication in 186 operations (80%), rest pain in 21 (9%) and gangrene in 24 (11%). The superficial femoral artery abnormality consisted of 1-10 cm occlusion in 52 cases (23%), > 10 cm occlusion in 96 (41%), single stenosis in 21 (9%), multiple stenoses in 28 (12%) and it was unknown in 34 cases (15%). Postoperative mortality was 0.8% with a complication rate of 10%. Five year cumulative primary patency was 71% overall, 75% in patients with disabling claudication, 61% in those with rest pain and 46% in those with gangrene. Eight year patency was 55% (S.E. 5.4%). No difference in 5-year cumulative patency was seen between treatment for stenosis or occlusion (74 and 70%, respectively). The results of earlier studies and the current study raise the question of whether endarterectomy should be the first treatment of choice in obstructive lesions of the superficial femoral artery. The results of endarterectomy are better than can be achieved with angioplasty and compare well with the results of femoropopliteal bypass. Endarterectomy has specific advantages above femoro-popliteal bypass: the autologous saphenous vein is spared for future use and in case of failure of the endarterectomy, femoro-popliteal bypass remains possible.(ABSTRACT TRUNCATED AT 250 WORDS)


A method of single-incision, four compartment fasciotomy of the leg. A method of achieving four compartment fasciotomy of the leg using a single skin incision is described. This procedure may be used to decompress the leg in acute compartment syndromes.


Verrucous carcinoma (epithelioma cuniculatum) plantare. A case of plantar verrucous carcinoma is described. The lesion was treated as an ischaemic ulcer before the true nature was recognised. Prior to the diagnosis the patient had a femoro-popliteal arterial bypass graft performed. An amputation was undertaken once the diagnosis was established.


Ruptured mycotic aneurysm of the abdominal aorta in childhood. Aortic aneurysms are exceedingly rare in childhood and, when mycotic, the risk of rupture is high. We report a case of ruptured mycotic abdominal aortic aneurysm presenting in a 9 year old girl, that was successfully repaired after initial misdiagnosis.


Fibromuscular dysplasia of visceral arteries. The case histories of two patients with combined renal and extra-renal fibromuscular dysplasia are presented. Readers are reminded of this rare but important condition as a cause of obscure abdominal symptoms and abdominal bleeding in the absence of atherosclerotic aortic aneurysm. The aetiology, histology, and management of the condition are discussed.


Delayed presentation of bilateral popliteal artery injury. We describe a patient who developed serious vascular complications following gunshot wounds to both popliteal fossae. There was minimal evidence of vascular injury on presentation to hospital, in particular ankle systolic pressures were normal. Five days following the initial injuries he was found to have a false aneurysm of the popliteal artery in his right leg and an arteriovenous fistula affecting the popliteal vessels of his left leg. The roles of arteriography and Doppler pressure studies in assessment of possible peripheral vascular injury following penetrating trauma are discussed. It is emphasised that a high index of suspicion and careful clinical review is essential if vascular injuries and their complications are not to be missed.


Chromosome preparation from human bone marrow--technique modified for immediate clinical investigation. A modified method for chromosome preparation from bone marrow aspirates of normal individuals and leukaemic patients is described for immediate clinical investigation. Hanks' balanced salt solution at different concentrations was used for in vitro incubation and hypotonic treatment. The method yields quite a high mitotic index with good metaphase spreads.


Quantification of neutral red pinocytosis by small numbers of adherent cells: comparative studies. A colorimetric assay has been developed for studies on neutral red (NR) pinocytosis by small numbers (below 2 x 10(5)) of adherent cells cultured in 96 well plates. The NR uptake per cell mass was much higher in the sea urchin perivisceral adherent cells and human HL-60 cell line monolayers than in the murine and Atlantic salmon macrophages. The apparent difference points to the usefulness of this novel assay in comparative studies.


Properties of acid beta-N-acetyl-D-glucosaminidase from cock semen. Two forms (I and II) of beta-N-acetyl-D-glucosaminidase from cock seminal plasma and one form (III) from spermatozoa were separated by chromatofocusing. The active enzyme forms I and II had pI values of 6.6 and 6.3, respectively, while form III had two subforms with pI values of 6.3 and 6.1, as determined by polyacrylamide gel electrofocusing. The molecular weights were 76,000 for forms I and III and 32,000 for form II. The optimum pH of enzyme forms I and III ranged from 3.6 to 4.0. In contrast, form II showed one distinct maximum at pH 3.7. The Km values obtained with p-nitrophenyl-beta-N-acetyl-D-glucosaminide as substrate were 0.35, 0.28, and 0.39 mM for forms I, II, and III, respectively. It is assumed that both cock spermatozoa and cock seminal plasma contain a common, enzymatically active beta-N-acetyl-D-glucosaminidase subunit with M(r) about 32,000 and pI 6.3.


Toxic effect of lead on fertility of inbred strain mice. Males of inbred strains were orally given lead acetate water solution during 150 days. Changes were found in the structure of the seminiferous tubules and in spermatozoa. The pathological changes in Leydig cells brought about a decline in levels of androgens.


Down-regulating effect on cellular immunity of the NSF17 factor produced by murine neonatal spleen cells fused with BW 5147 thymoma cells. The non-specific factor NSF17 produced by mouse newborn splenocytes fused with BW 5147 thymoma cells was tested in vivo for its immunosuppressive activity. NSF17 administered i.p. in a single dose into adult mice significantly decreased the PHA-induced proliferative response of lymphocytes, regional graft-versus-host reaction, and delayed-type hypersensitivity.


The influence of peat extract on the seminiferous epithelium of mouse testes. Because of the interest in the peat extract as a potential therapeutic agent, its effect on the seminiferous epithelium cells was studied. Adult male mice were intraperitoneally injected with peat extract during 34 days. At intervals equal to the duration of the cycle of the seminiferous epithelium (every 8.5 days), gametogenic cells were quantitatively analysed. It was revealed that the peat extract causes a decrease in the production of the A1 spermatogonia, and as a result a decrease in the intensity of spermatogenesis. Besides, in some individuals disturbances of meiosis took place, leading to an increased degeneration of pachytene spermatocytes and formation of diploid spermatids.


Effect of prolonged administration of insecticide (Cyhalothrin/Karate) on the blood and liver of rabbits. A sublethal dose of Karate administered to rabbits produced a significant increase in the total erythrocyte count and packed cell volume after 15 days of administration, though no significant change was observed after 30 days. The transaminases (glutamate oxaloacetate transaminase, GOT; glutamate pyruvate transaminase, GPT) also increased after 15 days of treatment. The GPT activity increased 119% and 60% after 15 and 30 days, respectively. From amongst metabolites, glucose content increased 17% and 185%, while cholesterol decreased 40% and 66%, and bilirubin 84% and 61%, after 15 and 30 days, respectively. The hepatic AkP activity decreased 30%, while the GPT activity increased 44%. Other enzymes such as AcP, GOT and LDH remained unaffected. The concentration of other metabolites, except for FAA which increased 35%, remained unaffected. Histological changes were marked by atrophied hepatic cells and hypertrophied nuclei and nucleoli. A trend towards necrosis of hepatic cells was also observed. All these results indicate that Karate is moderately toxic to mammals.


The prehospital use of nebulized albuterol on patients with wheezing whose chief complaint is shortness of breath. The use of prehospital nebulized beta-agonists has become widespread, and their safety and efficacy has been documented. Our purpose was to study their broadened use and determine their effectiveness in specific sub-sets of wheezing patients. We conducted a six and one-half month prospective study to determine the benefit of nebulized albuterol treatments on a variety of wheezing patients whose chief complaint to paramedics was shortness of breath. Sixty-two patients were enrolled in the study and were subdivided into four groups based on patient history; asthma, COPD, asthma & COPD (A/C), and non-Asthma/non-COPD (NANC). The effectiveness of the treatment was evaluated objectively by peak expiratory flow rates (PEFR) obtained before and immediately after treatment and subjectively by the patients' evaluation of their own dyspnea. Changes in PEFR were subjected to analysis by a paired T-test. Albuterol was effective in increasing the PEFR in patients with asthma, COPD and NANC. Patients with both asthma and COPD did not demonstrate increased PEFR after treatment. The majority of all patients were subjectively improved after nebulized albuterol treatments. We conclude that aerosolized albuterol is safe and effective in the prehospital treatment of patients complaining of dyspnea who are wheezing.


Is better access to medical care needed in Delaware? Improved access will require more primary care providers, a reduction in administrative red tape, and a clientele with healthy lifestyles who know how to use the system appropriately. Prevention is a necessary component of access even though it often takes years to produce results and is usually not dramatic or glamorous (the heart attack which does not occur is hard to measure). Over time, the health of Delawareans should show improvement through appropriate access to medical care, which should not only be cost effective but add to the quality of life.


Rheumatic fever: a case report.

Rheumatic fever: a case report. The case of a previously healthy nine-year-old female who presented with a two-week history of fever and five days of joint pain is discussed. She was admitted to the hospital, where the diagnosis of acute rheumatic fever was made. Although this diagnosis is made infrequently, it should be part of the differential diagnosis of persistent fever in patients presenting to the Emergency Department.


Outcome and long-term effects of pregnancy in women with hyperprolactinaemia. Twenty-four women with high circulating prolactin became pregnant on 39 occasions, of which 32 ended in delivery. Sixteen patients showed radiological evidence of pituitary tumour, 6 exhibited a normal CT and 2 had an empty sella. The pregnancies were induced in 4 patients after successful pituitary surgery, in 3 after surgery and medical treatment, and in the rest by bromocriptine (16) long-acting repeatable bromocriptine (1) and methergoline (1). No major complications related to hyperprolactinaemia or its treatment were observed during pregnancy in the patients or offspring. Prolactin after pregnancy was lower than before (basal 95 micrograms/l, after 1st pregnancy 38 micrograms/l P < 0.002, after 2nd pregnancy 24 micrograms/l P < 0.005 compared to basal prolactin); this prolactin reduction tended to be greater in the 9 multiparous patients, but did not attain statistical significance, probably because the number of multiparous patients was too small. A new empty sella developed after delivery in 4 women and persisted in another 2, all of which were medically treated; prolactin fell in all 6 cases normalizing in 3; 4 of these patients had undergone two or more pregnancies. The mean period of follow-up from the last pregnancy was 41.6 months (8-101). These data suggest that pregnancy may hasten a tendency to spontaneous improvement of hyperprolactinaemia, and multiparity may be beneficial in this way.


Paradoxical ovarian stimulations in the use of LHRH analogs. In the case of in vitro fertilization, LHRH analogs are used to induce a hypophysial blockage before the phase of stimulation, via administration of exogenous gonadotropin. During in vitro fertilization attempts using LHRH analogs, the blockage is controlled after 14 days of treatment by measuring plasmatic estradiol and by pelvic ultrasonography. In this retrospective study, which concerned 1075 in vitro fertilization cycles, a paradoxical ovarian stimulation with LHRH analogs was observed in 93 cases (8.7%) with high estradiol levels and follicular growth (detected by ultrasonography), in spite of low FSH and LH levels. In 4 cases, a follicular puncture was performed, which made it possible to collect oocytes from which embryos were obtained, thus confirming the observed follicular growth and maturation. The most probable hypothesis explaining this phenomenon seems to be direct ovarian stimulation effectuated in vivo by LHRH analogs. This stimulation is only observed in certain patients, and, more frequently it seems, with certain LHRH analogs, which is probably due to a variation in the expression of ovarian LHRH receptors.


A comparative analysis of two preparations of human chorionic gonadotrophin. A biophysical analysis of two pharmaceutical preparations of human chorionic gonadotrophin (Endo and Profasi) demonstrated that both contained high hCG specific activity. HCG protein analysis by occlusion HPLC and SDS gel electrophoresis showed similar profiles for both products. Although the purified protein components of both preparations were similar their were differences in the contents of the unpurified preparations. The Endo preparation exhibited greater inter-vial variability and contained an unidentified aromatic substance which interacted with the desalting column.


Cytogenetic analysis of 1508 spontaneous abortions originating from south Slovakia. Knowledge with regard to the spontaneous abortion cytogenetics has been derived exclusively from populations of the highly developed countries; its full applicability to east-european populations living in the peculiar conditions of the former 'socialist countries' may be questioned. The consecutive series of 1912 spontaneous abortion products with identifiable embryonal tissue was collected in six maternal clinics in south Slovakia. 1508 specimens were set in culture. Among 926 (61.4%) successfully karyotyped abortions, 46.0% were chromosomally abnormal specimens. The proportion of the groups of abnormalities is similar to that in other large series, with the exception of a higher proportion of mosaics (14.3% of the abnormalities). The abnormality rate in the whole sample also lies within the range of values published. However, significant inter-regional variability has been established: the chromosome abnormality rate in the urban subsample is 47.8%, whereas the value was only 35.3% for rural southern districts. The anomaly rates are lower for all maternal ages and gestation phases in the country subsample. The possibility of the environmental diversity effect was discussed. No other major peculiarity of cytogenetic abortion characteristics can be determined in the population studied.


The prognostic significance of maternal serum CA125 measurement in threatened abortion. The prognostic predictive value of maternal serum CA125 measurement was investigated in 25 cases of threatened abortion. The women were non-smoker, had a ultrasonographically verified viable single fetus, and the gestational ages ranged from 7 to 12 weeks. Twenty-five healty pregnant women, with the same characteristics were used as the control group. The overall abortion rate was found to be 20% (5/25) in the study group. In serial measurements the mean serum CA125 level of the patients with an unfavorable pregnancy outcome was significantly higher than that of the patients with a favorable outcome. When the cut-off level of maternal serum CA125 was taken as > 65 U/ml in the first and > 60 U/ml in the second measurements of the study group, the risk of termination of the pregnancy by spontaneous abortion was 83.3% in the patients with elevated serum CA125 levels. No statistically significant difference was observed with respect to the duration of vaginal bleeding between the aborters and the patients with a favorable outcome. Nevertheless, when vaginal bleeding had been present for 3 days or more and there was high maternal serum CA125 activity, the abortion risk was found to be 100% (3/3). These findings suggest that the maternal serum CA125 measurement in threatened abortion can be useful to determine the extent of decidual destruction which is directly related to the outcome of pregnancy.


Carbon dioxide laser treatment of cervical dysplasia in teenagers. Over a 6-year period, 40 young women under 20 years of age with cervical intraepithelial neoplasia (CIN) grade I (58%) to grade III (10%) were treated by laser vaporisation or excisional conisation. This prevalence of CIN in young women (nearly 4% of laser-treated cases) underscores the need for detection of cervical abnormalities in all teenagers engaged in sexual relations. The colposcopically-guided carbon dioxide laser technique is the treatment of choice for vaporisation and conisation of intraepithelial lesions (extending to the periphery of the cervix in 20% of cases) and for vaporisation of associated condylomatous lesions of the cervix (75%), vagina (23%), vulva (48%) and/or anus (8%).


An unusual type of endometrial cancer, related to tamoxifen? Tamoxifen, which is increasingly being used in breast cancer patients, has been associated with an elevated frequency of endometrial carcinoma. To our knowledge not a single case of uterine serous papillary carcinoma (USPC) has been documented during tamoxifen treatment. No conclusions as to a causal relationship are yet being made, but if it is due to tamoxifen, we should advise a strategy for prevention, because this subtype is not as curable as endometrioid carcinoma.


Unsuccessful methotrexate treatment of a tubal pregnancy with a live embryo. Two cases with unsuccessful local and systemic methotrexate (MTX) therapy of tubal pregnancy with fetal heart rate activity are reported. The three modes of therapy, the first with local potassium chloride solution injection, the second with local MTX injection, and the third with systemic MTX injection, failed in the resolution of viable ectopic pregnancy. Therapy of MTX in cases of tubal pregnancy with demonstrable fetal heart rate beats, should be reconsidered.


Congenital absence of fallopian tube and ovary. Absence of one or both uterine tubes and ovaries is a finding that has rarely been described. Two such cases are presented. In one the anatomical abnormalities were discovered during investigations for primary infertility, in the other the absence was discovered at the time of laparoscopic sterilisation. There are two possible aetiologies. The first involves an asymptomatic torsion of one or both adnexes during adult life or childhood, or even before birth. Alternatively, the absence may be congenital, due either to a defect in the development of the entire Müllerian and Mesonephric systems on one side, or to a defect localised to the region of the genital ridge and the caudal part of the Müllerian duct.


Primary amenorrhea-galactorrhea with hyperprolactinemia and huge pituitary enlargement in juvenile primary hypothyroidism. We report a girl with juvenile primary hypothyroidism revealed by growth retardation and a syndrome of primary amenorrhea-galactorrhea with hyperprolactinemia and suprasellar pituitary enlargement. Resolution of the pituitary enlargement and the amenorrhea-galactorrhea syndrome occurred after thyroid hormone replacement. No similar observation has been reported earlier in juvenile hypothyroidism.


Antenatal uterine activity monitoring of women at increased risk of preterm labour. The aim of the study was to investigate the usefulness of antenatal uterine activity monitoring in the management of women at increased risk of preterm labour on the basis of a past history of preterm birth or mid-trimester abortion. Uterine activity was recorded every 2 weeks between 20 and 28 weeks gestation. Activity was considered to be increased if pressure changes > 15 mmHg were detected. Fifty-eight women had uterine activity monitoring. Of them, 39 had normal uterine activity. Nineteen women had increased activity and they were randomized to either a study group (9) where the findings were revealed to the clinicians caring for them or a control group (10) where the findings were not revealed. There was no standard regimen of management for the study group except that additional uterine activity monitoring was performed to provide feed back to the clinicians about their interventions. The sensitivity, specificity, positive predictive value and negative predictive value of uterine activity monitoring for preterm births and for preterm births before 32 weeks were disappointing. Uterine activity monitoring was not useful for predicting births prior to 32 weeks; most of these were preceded by prelabour rupture of the membranes. The pregnancy outcome of women with increased uterine activity was not better if clinicians were aware of that increased activity than if they were not.


Survival and long-term morbidity in preterm infants with and without a clinical diagnosis of periventricular, intraventricular hemorrhage. The prognosis of clinically diagnosed periventricular, intraventricular hemorrhage on the survival and long-term outcome of 169 infants delivered at 24 to 30 weeks gestation who survived more than 48 hours was evaluated. Periventricular, intraventricular hemorrhage was confirmed by ultrasound in 37.9% of the survivors. In this group the survival rate was 64% and the major handicap rate was 14.6%. In contrast, the survival of infants who did not have a clinical diagnosis of periventricular, intraventricular hemorrhage was greater than 90% with a major handicap rate of only 3.2% (P < 0.0001 and P < 0.03, respectively). Infants with clinical diagnosis of periventricular, intraventricular hemorrhage who were found to have grades 1-2 by ultrasound had the same survival rate as those without a clinical diagnosis of periventricular, intraventricular hemorrhage (90.5%), while infants with grades 3-4 had a survival rate of only 51.2% (P < 0.01). We conclude, that preterm infants who survive longer than 48 hours and do not have a clinical suspicion of periventricular, intraventricular hemorrhage, have an excellent prognosis. In these circumstances brain sonography can be deferred without jeopardizing the infants' health.