When are simple flashes and floaters ocular emergencies? One hundred and seventy patients presented to the Bristol Eye Hospital casualty department with flashes and/or floaters during a six month prospective study. A sight-threatening condition was found in 41 patients (24.1%), the most important of which was a retinal break (16.5%). Although there were no specific symptoms which could be correlated to an increased incidence of retinal breaks, those patients who complained of isolated uniocular floaters had an insignificant incidence of breakage, when compared to asymptomatic fellow eyes. Thus while the majority of patients with flashes and floaters do merit an urgent ophthalmological opinion, those who complain of a single, isolated floater can safely be reviewed as routine outpatients.


A comparison of prophylactic, topical and subconjunctival treatment in cataract surgery. Prophylactic subconjunctival antibiotics and steroids have been used in cataract surgery since the early 1950s. However despite their widespread acceptance, there has, to date, been no direct comparison of subconjunctival and topical routes of administration. We therefore carried out a prospective comparison of the effects of subconjunctival and topical administration of antibiotics and steroids in sixty patients undergoing elective, uncomplicated, extracapsular cataract extraction and posterior chamber lens implantation. The patients were examined by a single observer on the first and third post-operative days and two and six weeks following surgery. The observer was blinded to the patient treatment group. Significantly higher degrees of conjunctival injection and anterior chamber activity were noted in those who received subconjunctival injections compared with those who received topical treatment. There were no significant differences between the two groups in best corrected visual acuity, central corneal oedema, cystoid macular oedema, intraocular pressure and infection. We conclude that prophylactic subconjunctival therapy in uncomplicated cataract surgery is not necessary.


The uses and limitations of intraocular biopsy. Based upon the author's considerable experience of trans-scleral resection of malignant melanoma of the choroid, a technique has been developed for the biopsy of tissues of the posterior segment of the eye. Its use in the management of atypical malignancy posing diagnostic difficulty and in the investigation of selected case of acute retinal necrosis, uveitis and retinal pigment epitheliopathy is described. In 34 trans-scleral biopsies of choroid, RPE and in some cases, retina, an adverse result occurred in only one case, this it was thought being due to not including pars plana vitrectomy as part of the biopsy technique. Pars plana vitrectomy is now regarded as an integral part of this form of biopsy.


The safety and efficacy of topical norfloxacin compared with chloramphenicol for the treatment of external ocular bacterial infections. The Norfloxacin-Chloramphenicol Ophthalmic Study Group. Two hundred and forty-six patients with signs of acute bacterial conjunctivitis and/or blepharitis were randomised to receive either norfloxacin or chloramphenicol for one week in this double-masked parallel group study. Ninety-two per cent of the norfloxacin-treated patients and 93% of the chloramphenicol-treated patients were rated as either clinically improved or cured at the end of the treatment period. Based upon pre-treatment bacteriological cultures, 31.3% of the patients had significant bacterial infection of the lids and/or conjunctiva. All of these culture-positive patients were rated as either clinically improved or cured. Based upon post-treatment cultures, 72 of 82 strains of Gram-positive and Gram-negative bacteria were eradicated or suppressed following treatment with either norfloxacin or chloramphenicol. However six of 41 strains persisted for norfloxacin and four of 41 for chloramphenicol. Two norfloxacin-treated patients and three chloramphenicol-treated patients had adverse experiences, predominantly ocular discomfort, which required cessation of drug therapy. Norfloxacin appears to be an effective and relatively safe agent for the treatment of bacterial infections of the lids and/or conjunctiva. In this study, norfloxacin was clinically and microbiologically similar in activity to chloramphenicol.


Cystoid macular oedema following neodymium:YAG laser capsulotomy a prospective study. Neodymium:YAG laser capsulotomy was performed in 65 eyes of 65 patients because of vision loss due to posterior capsular opacification or wrinkling following uneventful extracapsular cataract extraction with intraocular lens implantation. Eyes with pre-existing macular pathology were excluded from this study. In all of the eyes the posterior capsule was successfully opened. None developed clinical or angiographic cystoid macular oedema. One eye developed retinal detachment nine months after capsulotomy.


Low contrast visual acuity changes in human immuno-deficiency virus (HIV) infection. A pilot study of low contrast visual acuity testing using Regan Charts has been undertaken in 34 patients seropositive for Human Immuno-Deficiency Virus and 20 normal control subjects. Low contrast visual acuities of the HIV (+) patients both with and without HIV retinopathy were found to be significantly lower than the age-matched controls (p < 0.01). This finding is probably attributable to pathology related to HIV in the visual pathways/Central Nervous System. Lowest contrast Chart (Chart C) was found to be a useful diagnostic tool for HIV retinopathy and presumed neuropathy.


Do computerised visual fields and automated optic disc analysis assist in the choice of therapy in glaucoma? The choice of therapy in patients with glaucoma is determined by the presence of damage in the visual field or optic disc or by change in the appearance of the optic disc or visual field over time. Computerised visual fields and automated optic disc analysis can assist in this determination. Automated optic disc analysis permits the measurement of neuroretinal rim area which is more closely related to the presence of pathology than cup/disc ratio. Simple ophthalmoscopic techniques may also be used to estimate the size of the neuroretinal rim area. Automated perimetry allows accurate diagnostic information to be obtained in a reliable manner. This permits easier diagnosis of glaucoma as well as easier long term follow up for change. The availability of automated techniques in ophthalmology has helped the clinician in the choice of therapy. The use of astute clinical judgement remains however of the utmost importance.


Biometry and auto-fluorescence of the anterior ocular segment in diabetics with and without autonomic neuropathy: a case control study. The biometry and auto-fluorescence of the anterior ocular segment were examined in a case control study of two populations of diabetics. Cases, with diabetic autonomic neuropathy, and controls without neuropathy (forty in each group) were selected from a community based diabetic population and matched individually for age, sex, and type of diabetes. Scheimpflug and auto-fluorescence anterior segment photography were performed and analysed by digital image analysis in a masked fashion. There were differences between the groups in axial lens thickness, front radius of curvature of the lens, anterior chamber depth and lenticular auto-fluorescence. These findings are discussed in the context of the known association between autonomic neuropathy and narrow angle glaucoma, and possible common mechanisms of tissue damage in diabetes.


Changes in light scatter and width measurements from the human lens cortex with age. Light scatter and width measurements of the anterior cortical layers of the human lens were made in 50 eyes of 50 subjects using computerised linear scanning densitometry of Scheimpflug images. It was demonstrated that the amount of light scatter increased with age in all of the three major zones and that zone C3 showed the most marked increase. Most lens growth occurs in zone C2 with C3 showing little increase in width once it has become established. Zone C1 showed a tendency to decrease in width with age. In addition it was shown that the C3 zone, which is not present at birth and during early childhood, appears as a distinct layer during the second decade of life. Its scattering properties continue to increase throughout life, exceeding all other zones after approximately 30 years of age, in the absence of cataract. Possible explanations for the lens zone pattern are discussed.


Clinical comparison of automated and manual keratometry in pre-operative ocular biometry. Corneal measurements, using the manual (Topcon OM-4) and automated (Canon RK-1) keratometers was performed on 104 eyes of 104 patients undergoing cataract and implant surgery to assess the role of automated keratometry in pre-operative ocular biometry. Four eyes of four patients were excluded from statistical analyses for various reasons. The time taken to perform automated keratometry was a mean of 61 (SD 21) seconds compared to 205 (SD 37) seconds for manual keratometry; the difference was statistically significant (p < 0.001). In terms of the various keratometry values compared, mean K (corneal refractive power), flattest K, steepest K, astigmatism and the axis of astigmatism, 65% to 75% of the cases on automated keratometry were within 0.26 dioptres or 11 degrees of manually determined values; the difference was statistically significant (p < 0.001). Although automated keratometry was significantly quicker than manual keratometry, we continue to use manual keratometry values for intraocular lens power calculations as the accuracy demonstrated by automated keratometry was considered inadequate for this purpose.


Keratometry and post-operative astigmatism. Total post-operative refractive astigmatism and keratometric corneal astigmatism were determined in patients following cataract surgery. By comparing the results of the two methods, the value of keratometry as a simple and quick procedure for identification of surgically induced astigmatic errors was established. It is proposed that this method should be used before final refraction in an attempt to increase clinic efficiency and reduce further appointments resulting from surgically induced astigmatism.


Diabetes mellitus and retinal vein occlusion in patients of Asian, west Indian and white European origin. The incidence of retinal vein occlusion (RVO) is known to be related to several cardiovascular risk factors including diabetes mellitus, hypertension and hyperlipidaemia. We have assessed the prevalence of these risk factors in Caucasian (N = 536, mean age = 65.21 years), West Indians (N = 24, mean age = 57.3 years) and Asian (N = 28, mean age = 51.4 years) patients presenting with RVO. We found no significant differences between the three groups in the distribution of hyperlipidaemia, but diabetes mellitus was more common amongst both the Asians and W. Indians (10% v 29% and 38%, p < 0.01 and p < 0.001). Hypertension was also more common in both the Asians and the West Indians (59% v 64% and 83% m p < 0.001 and p < 0.0001). The Asians and West Indians presenting with RVO were significantly younger and had significantly higher body mass index than their Caucasian counterparts (Ethnic origin, BMI, mean age: Asian, 28.1 +/- 4.3, 51.5 +/- 12.3 years; West indian: 30.2 +/- 7.7, 57.3 + 10 years; White: 24.8 +/- 4.2, 65.2 +/- 11.4 years). In this study RVO occurred at a younger age in Asians and West Indians, and was associated with a greater prevalence of diabetes mellitus and hypertension. The prevalence of hyperlipidaemia was no different in the three groups.


Mathematical modelling of retinal tear formation: implications for the use of heavy liquids. When force is applied to detached retina it elevates, stretches, and becomes irreversibly deformed. Finally it tears, and the timing and location of tear formation are determined by the distribution of stress within the retina. This stress distribution is dependent upon the retinal contour. We have tested this using retinas of freshly enucleated bovine eyes. A suture was attached to isolated retinal tissue using butylacrylate (Histoacryl) glue and traction was applied to this suture so stretching the retina. We present a mathematical model of the in vitro data obtained, with predictions for local retinal internal tensions. We show that these are altered by the addition of hydrostatic forces due to the presence of heavy liquids. These findings have implications for epiretinal membrane dissection during surgery and for the use of heavy liquids during membrane peeling.


Genetic counselling in X-linked ocular albinism: clinical features of the carrier state. Forty-nine obligate heterozygotes and 64 at risk females were assessed from 17 families affected by X-linked ocular albinism to determine the relative values of clinical examination and skin biopsy in carrier detection. 92% of obligate heterozygotes had a mud-splattered appearance of the fundus with hyperpigmented streaks and in 74% this was associated with marked iris translucency. Skin histology showed macromelanosomes in 84%. 27 of 64 (42%) at risk females had definite features of the carrier state and 19 (30%) were unequivocally normal but a further 18 (28%) had mild RPE abnormalities, often with iris transillumination defects, of uncertain significance. Fundus examination will detect most carrier females but will not allow reliable genetic counselling for all at risk females, even when supplemented by skin biopsy.


Echographic diagnosis of advanced Coats' disease. Standardised echography was utilised to diagnose two cases of advanced Coats' disease. The characteristic acoustic features included extensive, poorly mobile retinal detachment; dense, dispersed subretinal opacities; looping of peripheral retina and absence of solid mass lesion and calcification. As enucleation of two eyes was necessary, it has proved possible to correlate the echographic and histological findings of this disease. The echographic features of other common causes of leukocoria are also discussed.


Ophthalmic consequences of mid-facial trauma. Fractures of the mid-face are commonly accompanied by injury to the visual system. Three hundred and sixty three patients who had sustained mid-facial fractures were assessed prospectively for evidence of such injury. The data have been used to devise a scoring system for the maxillofacial surgeon in order to help identify those patients at risk of eye injury. The scoring system has been evaluated on a further cohort of 100 patients with a similar spectrum of injury and the sensitivity and specificity of the system have been determined. The results of these studies have been reported in the maxillofacial literature. This paper reviews the data and results obtained. In summary, impaired visual acuity with a comminuted or out blow fracture, a motility abnormality, or facial fracture combined with head injury, sufficient to cause both retrograde and post-traumatic amnesia, emerged as major risk factors which are indicative of an adverse ophthalmic outcome deemed to warrant referral. The scoring system which was developed from this data was found to have a sensitivity of 94.4% and a specificity of 89% for the detection of patients thought to merit ophthalmic assessment. Failure to assess central visual function as objectively as practicable in patients who have sustained mid-facial fractures may lead to potentially treatable ophthalmic pathology not being identified.


Choroid plexus papilloma. A new presentation of von Hippel-Lindau (VHL) disease. A definite association between Von Hippel-Lindau [VHL] disease and choroid plexus tumour has not been described previously. A 24-year-old patient was found to have a choroid plexus tumour in the left cerebellopontine angle and involving the temporal bone. Examination of her fundi revealed bilateral retinal angiomatosis, thus making a diagnosis of von Hippel-Lindau disease. Later, an abdominal scan showed renal and pancreatic cysts. An important point is that molecular analysis of the choroid plexus tumour tissue showed chromosome 3 allele loss as described for other tumour types associated with von Hippel-Lindau disease.


Quantification of the ocular reactions to microfilaricides in the chemotherapy of onchocerciasis. Severe adverse systemic and ocular reactions have complicated the chemotherapy of onchocerciasis. A method for the quantification of ocular reactions to chemotherapy has been devised at the Onchocerciasis Chemotherapy Research Centre, at Hohoe, in Ghana. Symptoms, visual function, anterior segment inflammation and disease of the posterior segment are graded. The information is entered into a computerised database which allows reaction scores to be calculated both for individual patients, and for treatment groups. This system enables comparison of adverse ocular reactions to various new chemotherapeutic regimens.


Pyogenic granulomas of the lacrimal sac. Eight cases of pyogenic granulomas of the lacrimal sac are reported. The clinical symptoms were consistent with chronic dacryocystitis which was unresponsive to previous probing and antibiotics. Extirpation of the sacs was done because of dacryocystocoele, pyocoele, or fistula formation through the skin. Trauma and inflammation were thought to have contributed to the growth of the pyogenic granulomas. The histopathological examination revealed nonspecific granulomas consisting of well vascularised, friable tissue of a myxomatous nature containing an infiltration of lymphocytes, plasma cells, and a few eosinophils. Over the granulomas an epithelial layer of cells is stretched showing no signs of dysplasia or neoplastic growth. The term 'lobular capillary hemangioma' has been suggested to describe lesions morphologically similar to those observed in the study. These masses made up 53.3% of a total of 15 lacrimal sac tumour biopsy specimens examined over a period of 10 years (1979-1989). The few reported cases of primary lacrimal sac tumour of non-epithelial origin suggest that they make up a large minority of the lacrimal sac tumours. A review shows that relatively few of these lesions have been described in recent English literature but these growths are probably not being as consistently reported as primary epithelial tumours.


Immunology and immunopathology of reproduction. This review is dealing with the roles of the immune system in the development and functions of the male and female reproductive systems. Further, it describes the topical views on the roles of immunopathologic events and mechanisms involved in the male and female infertility, recurrent abortions, endometriosis, EPH gestosis and disorders of fetal development. Particular attention is paid to the complicated immunological cross-talk and interplay between the mother and its offspring, including the active role played by the placenta and mainly the trophoblast tissue, in the course of gestation. In the light of the "immunotrophic theory", maternal immune responses to foreign fetal components, occurring in normal pregnancies, within the limits of "tolerated" or even beneficial levels, are described. Emphasis is given on the possible deregulation of materno-fetal immunological balance, leading to immunopathological events and putting in danger the overall reproductive capacity of the couple. The contemporary therapeutic--mainly immunological--approaches to the main reproductive failures are also mentioned.


Autoimmunity: from physiology to pathology. Natural antibodies, mucosal immunity and development of B cell repertoire. Presence of spontaneously produced immunoglobulins bearing a broad spectrum of "natural" antibody specificities (including autoantibodies) in sera and other body fluids results mainly from inapparent immunization and polyclonal B cell activation by microflora and food antigens occurring mostly on mucosal surfaces. Early postnatal ontogeny in external environment is characterized by rapid growth and functional maturation of secondary lymphatic tissues as a consequence of this "natural" mucosal immunization. Under normal circumstances a state of "oral" tolerance to intestinal antigens is actively established after this period. Studies performed in germ-free, antigen-free and maternal antibody-deprived animals showed that low amounts of natural antibodies (mainly of IgM isotype) are formed without any known cause of stimulation. These "nonstimulated" antibodies, similarly as hybridomas originating from nonimmunized newborns, correspond to the preimmune repertoire of antibodies characterized by poly-specificity, high connectivity and reactivity against self antigens. Together with other innate humoral and cellular factors, they probably represent the first line of anti-infectious resistance. Moreover, due to their connectivity they are supposed to play an important role in B cell repertoire shaping (forming an idiotypic network), through interaction with a broad spectrum of immunological components they act as regulatory molecules, and through their participation in catabolic events they can promote morphogenetic changes during fetal development. Beneficial therapeutic effects of nonspecific gammaglobulin (IVIG) application observed recently in patients with autoimmune diseases suggest that they can influence autoimmune reactivity by a not yet analyzed mechanism. Other functions of natural autoantibodies can be suggested and expected to be found in the near future.(ABSTRACT TRUNCATED AT 250 WORDS)


IL-1 and pro-inflammatory cytokines produced by primary and transformed fibroblasts abrogate the tumorigenic potential of fibrosarcomas. In the present study we report on novel immunoregulatory functions lately attributed to fibroblasts, namely participation in cellular immune responses in connective tissues, by generation of pro-inflammatory cytokines and by presenting antigens to proliferating T cells. In order to execute immunoregulatory functions, the fibroblast has to be activated by signals abundant at inflammatory sites, i.e., cytokines and bacterial products. It was demonstrated that such immune-activated fibroblasts are able to generate a variety of cytokines such as interleukin-1 (IL-1), IL-6, colony stimulating factors (CSFs) as well as prostaglandins. The array of cytokines generated by immune-activated fibroblasts is determined by the stimulant and is controlled at multiple regulatory levels, such as transcription, translation, post-translational modifications, compartmentalization within the producing cell as well as the timing of expression. Some oncogene-transformed fibroblastoid cells lines were shown to constitutively generate IL-1 (and not IL-1 beta), as evidenced by the continuous expression of specific mRNA and biological activity of the cytokine, associated to the cell membrane or located in the cytosol. When these IL-2 producing cell lines were injected into mice, they failed to generate established tumors or regressed following initial growth, possibly due to mounting the host anti-tumor specific immune responses in which cytotoxic lymphocytes (CTLs) predominate. In contrast, IL-1 non-producing tumor cell lines induced progressive tumors which ultimately killed the animals. However, IL-1 non-producing fibroblastoid cell lines shifted from an in vivo progressive to a regressive phenotype, following immune activation of the malignant cells in vitro with cytokines/LPS. Similarly, primary immune-activated fibroblasts also induced tumor regression, mediated by anti-tumor specific immune responses, when the fibroblasts were injected into the vicinity of the tumor. Thus, the importance of activated stromal cells on tumor development was emphasized. This situation is relevant to the development of malignancies, as tumor growth is often accompanied by a local inflammatory response. Thus, the induction of IL-1 and other pro-inflammatory cytokines expression by the malignant cells or by stromal cells, in the vicinity of the tumor, might be efficient for tumor eradication. These findings should serve as a basis for development of novel immunotherapeutical strategies for the eradication of solid tumors.


H-2 influence on ankylosing enthesopathy of the ankle (ANKENT). Joint disease, ankylosing enthesopathy (ANKENT) of the ankle, occurs naturally in mice. ANKENT depends on the genetical constitution, sex, and age, and its frequency varies among inbred strains of normal mice. We found the highest frequency of ANKENT in B10.BR (H-2k) males. The H-2k haplotype appears to be a relative risk factor, which increases the susceptibility to ANKENT. The aetiology of ANKENT is unknown but an involvement of microbial agents in the environment is supposed.


Elevated IgA salivary antibody levels to mycobacterial 65 kDa stress protein in patients with oral candidosis. Salivary IgA antibody levels to cytoplasmic protein extract of Candida albicans (CSE) and to mycobacterial 65 kDa stress protein (mSP65) were determined by ELISA in 25 patients with chronic atrophic oral candidosis and in 23 controls with clinically healthy oral mucosa. The results showed significantly elevated anti-CSE and anti-mSP65 antibody levels in patients when compared with healthy controls (P < 0.001). A statistically significant relationship was found between salivary antibody titres to CSE but not to mSP65 and C. albicans colony counts in patients with oral candidosis.


Childhood typhoid fever diagnosed in an urban U.S. setting. The majority of cases of childhood typhoid fever diagnosed in the United States occur in persons recently returned from travel abroad. We report two childhood cases of typhoid fever acquired in Mexico and India and diagnosed in an urban United States setting. This report describes these cases and provides insights into the pitfalls in recognizing this condition in a nonendemic setting as well as information concerning differential diagnosis and treatment.


Cesarean section in a severely traumatized patient. Massive trauma in a near-term pregnant patient challenges the emergency physician with very difficult treatment choices. We present the case of a severely traumatized pregnant female, near term, in whom the decision to proceed with a perimortem cesarean section in the Emergency Department produced a viable infant. We discuss the guidelines that are used in making such a decision and their rationale. The physiologic changes of pregnancy that may complicate maternal assessment and the effect of shock on the fetus are also discussed.


Census adjustment and the distribution of federal spending. In 1989, programs that use population counts to determine the distribution of their funds transferred $236 per capita to state and local governments. If the 1990 census were adjusted to reflect undercounting, about 40% of state and local governments would receive increased grants averaging $56 per miscounted person; other jurisdictions would lose an almost equal amount of grant money. The surprisingly small reallocations arise because 1) total funds allocated by population are essentially fixed; 2) allocations depend on other factors in addition to population; and 3) programs vary as to whether they allocate funds in direct or inverse proportion to population.


Age of entry into marriage and the date of the initiation of voluntary birth control. It is widely known that modern economic development has been accompanied by the initiation and spread of effective limitation of fertility, and that generally the populations which experienced development at a late date also had a belated reduction in childbearing. Here a surprising relation is found between (and within) broad regions: the areas in which traditional age of entry into marriage was late were the areas in which marital fertility was reduced first.


A simple measure of fertility control.

A simple measure of fertility control. We describe a simple measure of fertility control: the proportion of all births from the age-specific fertility schedule that occurs among women by age 35. This measure has broad applicability because it does not require information on marital fertility rates. When both the proportion of births by age 35 and the most commonly used measure of fertility control, m, are calculated for a population over time, they are correlated very highly. Because of increasing levels of nonmarital fertility in several developed countries, measures of fertility control that are based on marital fertility are less appropriate now than in the past.


The relationship between cohabitation and divorce: selectivity or causal influence? Recent evidence linking premarital cohabitation to high rates of divorce poses a complex theoretical and empirical puzzle. We develop hypotheses predicting that premarital cohabitation is selective of those who are prone to divorce as well as hypotheses predicting that the experience of premarital cohabitation produces attitudes and values which increase the probability of divorce. Using multiwave panel data from a recent cohort of young men and women in the United States, we specify and test models of these predictions. The results are consistent with hypotheses suggesting that cohabitation is selective of men and women who are less committed to marriage and more approving of divorce. The results also are consistent with the conclusion that cohabiting experiences significantly increase young people's acceptance of divorce.


Feathered nest/gilded cage: parental income and leaving home in the transition to adulthood. The growing study of leaving home in young adulthood in the United States has been hampered by data and measurement problems, which are producing a major theoretical confusion about the role of parental resources in influencing young adults' leaving home. Does high parental income retain young adults in the home or subsidize their leaving (and parental privacy)? This paper uses the 1984 panel of Survey of Income and Program Participation to clarify this issue, and shows that the effects of parental resources differ depending on the route out of the home under consideration (marriage or premarital residential independence). Effects change substantially over the nest-leaving ages, but relatively few differences are found between young men and young women.


The effects of child health on marital status and family structure. The purpose of this paper is to provide evidence on the effect of child health on marital stability and family structure in an economic framework. We use the 1988 National Health Interview Survey's Child Health Supplement, with a sample of about 9,000 families, to test whether having an unhealthy child decreases the mother's chance of being married and whether it increases her chance of living in an extended family. Using two different measures of child health, we find that having an unhealthy child decreases the mother's likelihood of being married. Our results imply that children in poor health are more likely to face obstacles beyond their illness because they also are more likely to suffer the consequences of poverty and the poor schooling outcomes that result from being raised in a female-headed household. The only mitigating factor is that unhealthy white children are more likely than their healthy counterparts to be living in an extended family.


Drug use as a risk factor for premarital teen pregnancy and abortion in a national sample of young white women. The relationship between adolescent drug use and premarital teen pregnancy and abortion as a pregnancy outcome among sexually active women is investigated in a sample of white women from the National Longitudinal Survey of Youth. Event history analysis is used to explore whether prior drug use has a unique effect on premarital teen pregnancy, with controls for personality, lifestyle, and biological factors. Logistic regression is used to estimate whether drug use affects the decision to terminate a premarital teen pregnancy. The results show that the risk of premarital teen pregnancy is nearly four times as high for those who have used illicit drugs other than marijuana as for those with no history of any prior substance involvement. Furthermore, illicit drug use increases the likelihood of an abortion by a factor of 5. Policy implications of the findings are discussed.


Why are teenagers in the United States less likely to breast-feed than older women? Teenage mothers are much less likely than older mothers to breastfeed their infants. The lower breastfeeding rate among teenagers aged 16-19, compared with women aged 20-29, is due almost entirely to the fact that teenage mothers tend to have characteristics associated with a lower likelihood of breastfeeding among all women, such as lower educational level, lower income, and being unmarried. Even so, nearly 40% of the difference between teenage mothers aged 15 or less and mothers aged 20-29 remains unexplained by these factors and may be due to developmental aspects of adolescence, such as greater egocentricity and greater concern about body image.


Residential preferences and residential choices in a multiethnic context. A study of the expressed preferences of four different ethnic groups in the Los Angeles metropolitan area shows strong desires for own-race combinations in the ethnicity of neighborhoods that individuals say they would choose when seeking a new residence. The results also show that Anglos are not the only group to practice "avoidance" of other racial/ethnic neighborhoods, although avoidance behavior by Anglos is the strongest. Because the issues of racial composition are socially sensitive, additional tests examined the relationship of preferences to behavior. Although many behaviors generally follow expressed preferences, members of households who expressed "no preference" also were found to largely choose own race neighborhoods. The results of this study suggest that the expressed preference for own race/own ethnicity, in combination with short-distance local moves, is likely to maintain present patterns of separation in U.S. metropolitan areas.


Immunogenetic and evolutionary influences on the host-parasite relationship. Immunogenetic aspects of the host-parasite relationship between the laboratory mouse and the parasitic nematode Trichinella spiralis are reviewed. Variation in the capacity of hosts to express an effective protective immunity is associated primarily with variation in inflammatory responsiveness. Analysis of the contributions of T lymphocytes, cytokines, and myeloid precursor populations to variation in one particular inflammatory component, eosinophilia, is described. Variation in parasite immunogenicity at the level of host responsiveness to different parasite isolates is described briefly. Variations in host-protective immunity, immunopathology, and parasite immunogenicity are discussed in terms of evolutionary pressures selecting for optimal host and parasite survival in a stable symbiosis.


Correlation of subjective assessment of amniotic fluid with amniotic fluid index. We assessed the correlation between abnormal amniotic fluid volumes as defined by the two techniques of (1) subjective evaluation and (2) the amniotic fluid index. Ultrasound evaluation of amniotic fluid volume was conducted on 420 pregnant women with known gestational age greater than twenty weeks but less than 42 weeks. Amniotic fluid was evaluated subjectively and placed into one of three categories: normal, oligohydramnios or polyhydramnios. After fetal biometry was performed, the amniotic fluid volume was assessed semi-quantitatively by the amniotic fluid index technique and assigned to similar categories. We analyzed the data with 2 x 2 contingency tables, using amniotic fluid index as the 'gold standard test'. Our study demonstrates that there was moderate agreement (kappa.5) between both amniotic fluid techniques in the identification of oligohydramnios. However, agreement between the techniques was poor for the identification of polyhydramnios (kappa.16).


Function associated molecules on nonspecific cytotoxic cells: role in calcium signaling, redirected lysis, and modulation of cytotoxicity. Monoclonal antibodies (mAbs) specific for a putative antigen receptor (function associated molecule, FAM) on fish NCC were tested for their ability to initiate signal transduction responses. Anti-FAM hybridoma target cells were significantly lysed by NCC when tested by redirected lysis (RL). Redirected lysis was enhanced by 1-h pretreatment of the NCC with anti-FAM mAb; however, 18-h pretreatment produced significant inhibition. The effects of the calcium ionophore A23187 on RL were also examined: 10(-4) M but not 10(-5) M A23187 significantly increased RL. Purified NCC were treated with mAb and cellular DNA synthesis was determined. Significant increases in NCC incorporation of tritiated thymidine were measured 72 h after treatment with mAb. Experiments were also conducted to determine levels of free cytosolic calcium in NCC following mAb binding. Anti-FAM mAb binding produced a rapid rise in cytosolic calcium (200-500 nM) determined by Fura-2/AM fluorescence. Calcium ionophore A23187, PHA, and Con-A treatment of NCC also produced significantly increased [Ca2+]i. Target cell binding by NCC caused a three-fold increase in [Ca2+]i. These data suggest that triggering of the FAM may initiate a signal(s) that activates cytotoxicity, increases cytosolic free calcium concentrations, and initiates DNA synthesis.


A species-conserved NK cell antigen receptor is a novel vimentin-like molecule. The role of a novel, evolutionarily conserved function-associated molecule (FAM) in NK cell function has been examined in several species. This molecule has previously been shown to mediate NK and NK-like recognition functions in fish NCC and human NK cells. We now show that this molecule is distributed in those tissues that contain NK cells in mice and rats. Further, we show that this molecule functions as an antigen receptor on NK cells of these species. That is, monoclonal antibodies directed against this FAM inhibit NK cell cytotoxic function and trigger signal transduction pathways in each of the species. Finally, we present evidence that this putative antigen receptor is a vimentin-like molecule which functions to mediate all NK or NK-like recognition functions in a variety of species.


Loss of PAF-like activity from human embryo conditioned media (ECM) following HPLC separation. Recently a platelet activating factor (PAF)-like activity has been found in embryo conditioned media (ECM) and consequentially been termed embryo-derived PAF (EPAF). Yet it remains unclear whether the embryo-released molecule is in fact PAF or a PAF precursor or inductor in vivo. In this study we shall show that ECM did not induce platelet aggregation in vitro; however, it was possible to detect PAF-activity using the sensitive splenectomized mouse bioassay (SMB). Following lipid extraction, PAF activity was diminished, and after additional HPLC separation completely lost. We propose that the active fraction of ECM is lipid in nature but that this molecule is not PAF. We would rather suggest that this molecule induces the production of PAF by other cell types in vivo.


NADPH oxidase-like activity in rainbow trout Oncorhynchus mykiss (Walbaum) macrophages. Evidence for the existence of an NADPH oxidase-like enzyme in rainbow trout macrophages is given. Reduced-minus-oxidised difference spectroscopy revealed the presence of a cytochrome b with three absorbance peaks, at 430, 533, and 558 nm. The low midpoint potential of the latter peak suggests this cytochrome is the same as the terminal component of NADPH oxidase (i.e., cytochrome b-245). Subcellular fractionation of macrophages revealed two peaks of cytochrome b activity, in accord with the concept of a plasma membrane localisation of cytochrome b activity in addition to a mitochondrial localisation. Finally, that the rainbow trout oxidase is a multicomponent enzyme was suggested by inhibitor studies, where specific inhibitors of the flavin and cytochrome b-245 components of NADPH oxidase induced significant reduction in superoxide anion production.


Conservative management of placental abruption complicated by severe clotting disorders. Placental abruption resulting in fetal death may seriously affect maternal health, especially when clotting disorders arise. The prevailing view is that the clotting system will only normalize after the uterus has been evacuated and therefore prompt delivery, often by induction of labor, is advocated. Over a 3-year period, 16 cases (0.35%) of intra-uterine fetal death due to placental abruption occurred at Leiden University Hospital. Five cases were complicated by severe clotting disorders. All women delivered vaginally. No maternal deaths occurred. In contrast to most authors, we present data showing that in four out of five cases the clotting system started to normalize before the uterus was emptied. We suggest that an expectant approach until recovery of the coagulopathy is safer than aiming at a quick delivery. After stabilization of the clotting disorder, one may then consider evacuation of the uterus, if need be by induction of labor.


Hysteroscopic diagnosis of ectopic pregnancy. Although vaginal ultrasonography combined with plasma beta-hCG determination can provide a reliable diagnosis and location of ectopic pregnancy, the results can be difficult to interpret in the early stages when hCG levels are low. Hysteroscopy can be used in such cases to differentiate between ectopic pregnancy and non-viable uterine pregnancy when viable uterine pregnancy has been ruled out. General anaesthesia and laparoscopy are avoided. We performed 60 hysteroscopic procedures between January 1989 and December 1990 in patients with suspected ectopic pregnancies. The pregnancy had been located by means of vaginal ultrasonography in every case in which the hCG was above 1500 IU/ml and in 36% of cases in which the beta-hCG was below this level. Hysteroscopy was hindered by metrorrhagia in three cases and was inconclusive in one, necessitating laparoscopy. Diagnosis was possible in all the remaining cases, as follows: ectopic pregnancy in 41 cases, with an empty uterus and occasional bleeding from an ostium; non-viable uterine pregnancy in 18 cases, with the presence of material within the cavity. Hysteroscopy therefore confirmed the diagnosis in 55% of the cases and was itself diagnostic in a further 43% of cases. Its sensitivity for the diagnosis of ectopic pregnancy was 100% and its specificity 95%. We propose a diagnostic decision tree.


A peculiar immunoglobulin M (IgM) identified in eggs of chum salmon (Oncorhynchus keta). A protein reacting with antibody against chum salmon serum IgM was found in salmon egg yolk. The protein tentatively named IgM-like protein was partially purified from egg yolk extract by removal of euglobulin from the extract by dialysis against a low ionic strength buffer following DEAE ion-exchange chromatography. The IgM-like protein was antigenically identical with serum IgM, showing a complete fusion of precipitin line of the IgM-like protein with that of serum IgM on double immunodiffusion with antiserum IgM. The molecular weight of intact IgM-like protein was assessed by 3% SDS-PAGE to be 495 kDa, smaller than that of serum IgM (750 kDa). Upon 10% SDS-PAGE with mercaptane and subsequent Western blotting with antiserum IgM, the IgM-like protein separated into three components with molecular weights of 68 kDa, 51.5 kDa, and 23 kDa. The 68 kDa, the most minor component, and the 23 kDa, the smallest molecular weight component, were identified, respectively, as H and L chain in view of their molecular weights identical to those of serum IgM. The 51.5 kDa, the main component, was also identified as H chain, since it reacted with antiserum IgM absorbed with purified L chain. From these results, it was concluded that the IgM-like protein in egg yolk, having a lower molecular weight than that of serum IgM, consists of H chain of smaller molecular weight as well as H and L chain of ordinary molecular weight.


Vaginal progesterone as luteal phase support in an IVF/GIFT programme. To determine whether luteal phase support with vaginal progesterone could improve pregnancy rates in our IVF/GIFT programme, we performed a prospective randomised controlled study. After stimulation with clomiphene citrate/human menopausal gonadotrophin, 123 women received no luteal support and 122 received progesterone pessaries 100 mg b.d. from 48 hours prior to embryo transfer and continued throughout the luteal phase. There was no difference in the pregnancy rate following IVF/ET (6/58 and 10/58 for the pessary and control group respectively), but a significantly higher rate was noted for GIFT (13/34 and 5/42 for the pessary and control group respectively; P less than 0.05). Of interest, only one of the 19 pregnancies using luteal support was extra-uterine, compared with 6/15 in the control group.


Platelet-activating factor levels in human follicular and amniotic fluids. Platelet-activating factor (PAF) was estimated in extracts of human follicular and amniotic fluids using a commercially available 125I-radioimmunoassay. Levels obtained before and after purification of the extracts by thin-layer chromatography were similar and PAF could be estimated with high accuracy and reproducibility over a wide dilution range. PAF levels in fluid aspirated from mature (17 mm average diameter) follicles from in vitro fertilization patients were 1005 +/- 129 fmol/ml in successful (clinical pregnancy) cycles and 949 +/- 75 fmol/ml in unsuccessful (failure of implantation) cycles. PAF levels were also similar in follicles containing eggs and those where no egg was found, and in fluid from follicular cysts characterised by low oestradiol and progesterone levels. PAF was detected in midtrimester amniotic fluid (60 +/- 20 fmol/ml). At term, PAF was low or undetectable in amniotic fluid obtained at elective caesarean section before the onset of labour, but increased to 102 +/- 28 fmol/ml in samples obtained following spontaneous labour and vaginal delivery. The data suggest that PAF may be involved in ovulation and parturition but further studies into the mechanism of action of PAF are necessary. The good performance of the radioimmunoassay and the convenience of gamma-counting are strong advantages of this method over the available bioassays, but its expense remains a drawback.


Developmental patterns of bioactive and immunoreactive FSH in the female rabbit: effects of ovariectomy. A longitudinal study was performed to determine the relationship between immunoreactive and biologically active FSH in the serum of sham-operated and ovariectomized female rabbits. Twenty-two-day-old female rabbits, 8 per group, were sham-operated or bilaterally ovariectomized on day 23. Blood was taken every 3-4 days from each rabbit until they achieved a weight of 3 kg or age 100 days. Sera were analysed by radioimmunoassay for LH and FSH or for bioactive FSH. In sham-operated animals, immuno-FSH levels showed a 10-fold increase from 0.36 +/- 0.04 ng/ml to greater than 35 ng/ml between days 45 and 80. By contrast, bio-FSH levels increased more gradually from a baseline of 5.4 +/- 0.4 ng/ml to about 8 ng/ml. Bilateral ovariectomy resulted in a significant increase in both bio-FSH, 5.5 +/- 0.4 ng/ml to 12.6 +/- 1.5 ng/ml and immuno-FSH levels from 0.4 ng/ml to 5.2 +/- 1.4 ng/ml 24 h later. These levels of FSH remained elevated throughout the sampling period in both groups of animals and then decreased after day 100. Peripheral LH levels showed much more variation but were 2-fold higher in ovariectomized rabbits, 0.4-1.4 ng/ml in sham-operated vs. 0.8-2.4 ng/ml in ovariectomized rabbits. These results emphasize the marked variations in FSH levels according to the method of analysis. They also suggest that extra-ovarian factors may play a role in inhibiting gonadotropin release especially LH.


Aspiration of ectopic pregnancy under guidance of vaginal ultrasonography. This case report describes the treatment of tubal pregnancy by aspiration of the gestational sac content and injection of methotrexate by transvaginal sonographic guidance. The confirmation of the embryo aspiration as indicated by disappearance of the embryo with its heart beat and the resulting 46,XY karyotype, raises the question whether methotrexate injection is necessary.


Short-term effects of ritodrine, aminophylline and atropine on umbilical artery blood flow velocity waveform. With the recent introduction of Doppler technology, a non-invasive methodology which enables to evaluate qualitative changes in circulatory vessels, it is possible to investigate the possible effects of various drugs on several parameters of utero-placental-fetal circulation. In the present study we evaluate the flow velocity waveform (FVW) of umbilical artery (UA) during and after administration of aminophylline, atropine and ritodrine to healthy pregnant women. In our study we did not observe any significant short-term variation of PI after the administration of these drugs. Slight variations were detected, and they may be interpreted on the basis of the mode of action of these drugs. Doppler technology may be a useful tool for monitoring some effects on the fetus of the maternal administration of therapeutic agents.


Gemeprost-induced cervical ripening: histological and biophysical effects. The mechanism of prostaglandin-induced cervical ripening is not clear. The aim of this study was to measure the biophysical and histological effects of gemeprost on the cervix. Thirty-four women admitted for surgical termination of pregnancy in the first trimester were randomised in a double-blind manner to receive either gemeprost or placebo prior to surgery. In 20 (10 active, 10 placebo) a needle biopsy was taken from the anterior lip of the cervix prior to cervical dilatation for histology. The forces required to dilate the cervix from 3 mm to 10 mm were measured. A group of seventeen parous women undergoing surgical termination who did not receive gemeprost were also studied as a parous control group. A needle biopsy was obtained in eight of them and also in six non-pregnant parous women undergoing hysterectomy for benign conditions, the latter acting as a non-pregnant control group. Polymerised collagen was stained with Picrosirius red and glycosaminoglycans with alcian blue using a MgCl2 gradient. Optical densitometry was used as an objective measure of staining. Neutrophil concentration was assessed immunohistochemically. Gemeprost treatment increased free passibility (the size of the largest dilator which could be passed without encountering resistance) (P < 0.01), reduced the forces required to dilate the cervix (P < 0.01) reduced blood loss (P < 0.05), reduced the collagen concentration (P < 0.01) and was associated with a modest neutrophil influx (P < 0.02) as compared to placebo. The pregnant parous group had a significantly lower collagen concentration than the non-pregnant parous group (P < 0.02).(ABSTRACT TRUNCATED AT 250 WORDS)


Intra-amniotic pressures following vaginal gemeprost prior to first and second trimester termination of pregnancy. Intra-amniotic pressures were measured following 1 mg gemeprost for cervical preparation before first trimester vacuum aspiration (n = 10) and following 2 mg gemeprost before second trimester dilatation and evacuation (n = 15). Twenty-five women, matched for gestational age and parity, who did not receive gemeprost served as controls. Compared to control values (2-8 mmHg), basal intra-amniotic pressure (IAP) was significantly increased after 1 mg and 2 mg of gemeprost (median 20.0, range 4-45 mmHg, median 20.0, range 8-60 mmHg, respectively). Uterine contractions were recorded in 8 of 10 subjects after 1 mg (median delta IAP 28.0, 95% CI 10.0-42.6 mmHg) and 14 of 15 subjects after 2 mg (median delta IAP 52.5, 95% CI 26.7-60.3 mmHg). Gemeprost produces an increase in uterine contractility which may be additional to cervical softening properties and which may be responsible for the adverse effects of pain and bleeding experienced by some women prior to termination.


Predictive value of pelvic scores for induction of labor by local PGE2. The predictive value of pelvic scores, parity, age and gestational age for induction of labor by local prostaglandin-E2 (PGE2) was examined in 336 women attempting induction of labor by intracervical or vaginal PGE2. The patient characteristics were correlated to: (1) vaginal delivery within 48 h, (2) the period from induction to onset of labor (latency period), and (3) the duration of labor. The Bishop score (P < 0.01) and even more the Lange score (P < 0.0001) were significantly inversely correlated to both latency period and induction-delivery period. This was caused by cervical dilatation (P < 0.001), fetal station (P < 0.05) and cervical length (P < 0.05), whereas position and consistency of the cervix were of no importance. All three periods studied were significantly (P < 0.0001) shorter in parous women. In primiparous women, gestational age was of no importance for the latency period; however, higher gestational age was associated with longer labor (P < 0.001). We conclude that the predictive value of pelvic scores on induction hardly differs using local PGE2 compared to conventional methods; furthermore, the Bishop score should be substituted, disregarding position and consistency of the cervix, but putting more weight to cervical dilatation. A new pelvic score is proposed.


Impaired 1,25-dihydroxyvitamin D production in pregnancy-induced hypertension. The aim of the study was to evaluate the calcium metabolism in pregnancy-induced hypertension. Fifty-three women with pregnancy-induced hypertension were studied and the control groups comprised 20 women with uncomplicated pregnancies in the third trimester and 51 non-pregnant women, respectively. The mean serum concentrations of 1,25-dihydroxyvitamin D in women with pregnancy-induced hypertension was low (38.6 +/- 21.4 pg/ml) compared to women with uncomplicated pregnancies (91.0 +/- 18.2 pg/ml), but comparable to levels in non-pregnant women (32.2 +/- 11.9 pg/ml). Mean serum levels of PTH and ionized calcium were comparable in women with pregnancy-induced hypertension and women with uncomplicated pregnancies. In conclusion, the calcium metabolism in pregnancy-induced hypertension was changed compared to uncomplicated pregnancies with respect to the serum concentration of 1,25-dihydroxyvitamin D.


Blood flow velocity in the fetal abdominal aorta and in the umbilical artery in uncomplicated pregnancies. The blood flow velocity waveform (BFWV) in fetal vessels depends on the stroke volume and the frequency of the fetal heart, the compliance of the vessel and the peripheral resistance of the fetal vascular bed. The objective of the present study was to establish the change of the BFVW throughout gestation and whether the change of the resistance indices are related to the peak flow velocity and/or the end-diastolic flow velocity. The BFVW of the fetal abdominal aorta and of the umbilical artery at 27th-30th week and at 37th-40th week of gestation were analysed in fifteen patients with uncomplicated pregnancies and subsequent normal outcome. The measurement was performed with a pulsed duplex scanner (Kranzbühler). The Doppler beam had a fixed angle of 50 degrees to the fetal vessel in all cases. The peak flow velocity in the fetal aorta remained constant from the 27th-30th week to the 37th-40th week of gestation: 1418 +/- 248 Hz vs. 1448 +/- 269 Hz. The end-diastolic flow, however, showed a significant increase during the respective periods: from 270 +/- 59 Hz to 325 +/- 69 Hz. The peak flow velocity in the umbilical artery was about 25% below that of the fetal aorta: 1028 +/- 149 Hz (27th-30th week) and 1106 +/- 196 Hz (37th-40th week). The end-diastolic flow, however, increased by about 41%: 292 +/- 86 Hz vs. 412 +/- 83 Hz demonstrating a tremendous change of the compliance and the peripheral resistance in the umbilical vascular system. These alterations are also exhibited by the resistance indices. The resistance indices S/D, PI and RI of both vessels were related to the EDF of the abdominal aorta and the umbilical artery at the respective gestational age. It demonstrated that the EDF is of major influence on the calculated resistance indices. The blood flow in the aorta and the umbilical artery was 135(+/- 18) ml/kg/min and 143 (+/- 21) ml/kg/min, respectively. In conclusion, the BFVW for fetal surveillance should be measured under a constant angle in the fetal abdominal aorta. It gives a good information concerning the peripheral resistance and compliance in the respective vascular areas. The EDFV has its lowest value at 200 Hz in the fetal aorta and in the umbilical artery under physiological conditions.


The relationship between prior cervical conization, cervical microbial colonization and preterm premature rupture of the membranes. The occurrence of prior cervical conization and the cervical microbial colonization was investigated in 38 women with idiopathic preterm labor, 35 women with preterm premature rupture of the membranes (PPROM) and 75 normal pregnant women at 26-34 weeks of gestation. Data were analyzed by Fisher's exact test (two-tailed). The frequency of prior cervical conization was significantly higher in PPROM patients compared to normal pregnant women (P < 0.001) and to patients in preterm labor (P < 0.01). Lactobacilli occurred with a lower frequency in patients with PPROM compared to patients in preterm labor (P < 0.05) and control patients (P = 0.0543)-and with a lower frequency in patients with prior cervical conization (P < 0.05). All other microorganisms occurred with the same frequencies in all groups. The absence of lactobacilli may indicate changes in the cervical flora, which could increase the risk of PPROM. Prior cervical conization may impair the antimicrobial defense-mechanisms in the cervix, which could facilitate ascending microbial colonization. This may lead to a release of prostaglandins and proteolytic enzymes and subsequently preterm labor and rupture of the membranes.


Pregnancy after fifty: profile and pregnancy outcome in a series of elderly multigravidae. A series of seven women aged 50 years and over was studied over two years to analyse patient profile and pregnancy outcome. All were first-generation immigrant Asian multigravidae who were Muslims of low socio-economic status and who had never used contraception. The median age at booking was 52 (range 51-59 years) and the median parity was nine. One woman had gestational diabetes and one had an antepartum haemorrhage. There were few intrapartum or puerperal problems but an increased rate of Caesarean section (28.6%) was noted. There were no stillbirths, neonatal deaths or congenital anomalies in this series. The mean birth weight was 3.3 kg and the mean gestation at delivery was 39.2 weeks. There was no significant perinatal morbidity. Five of the seven women declined further family planning advice. A possible relationship between race, parity and age at menopause is discussed.


Prophylactic oophorectomy.

Prophylactic oophorectomy. The risks and benefits of prophylactic bilateral oophorectomy (BO) accompanying hysterectomy are reviewed. The potential reduction in the risk of subsequent ovarian cancer is quantified using literature data and a mathematical model. The risks of subsequent breast cancer from low-dose ERT are estimated using a comprehensive mathematical model. It is hypothesized that BO may have a substantial protective effect on breast cancer risk despite subsequent low-dose or non-low-dose ERT, when BO is performed at an early age. In women with a family history of ovarian cancer or breast cancer, the individual risks must be assessed on the basis of a pedigree analysis. In conclusion, a decision on BO must be based on weighing the potential benefits of reduced ovarian and breast cancer risks against the psychological importance of retaining the ovaries and the risk of osteoporosis and cardiovascular disease when compliance with ERT is less than perfect. The provided quantitative data may help in making the right decision.


Lack of effect of topical iodostearic acid on cervical intraepithelial neoplasia II-III. Stearic and iodostearic acid inhibit growth of a cervical carcinoma cell line in vitro. This study was performed to determine if iodostearic acid would induce regression of cervical intraepithelial neoplasia (CIN). Women with histologically-proven CIN II or III were randomised into two groups. Those in the first group were given pessaries composed of iodostearic acid in polyethylene glycol (PEG) base. Women in the second group were given pessaries containing only the PEG base. One pessary was inserted into the vagina nightly for 30 nights, and each woman then had the CIN lesion removed by CO2 laser cone excision. There was no difference in the histology of the cone biopsies between the groups, demonstrating that this regime of iodostearic acid has no useful role in the treatment of CIN II-III.


Management of eclampsia: cardiopulmonary arrest resulting from magnesium sulfate overdose. An eclamptic seizure occurring at 32+2 weeks of gestation was treated with magnesium sulfate. Accidentally an overdose was given. As a consequence, the patient had a cardiopulmonary arrest. Immediate resuscitation and calcium gluconate administration did restore vital functions. Pregnancy continued for 4 weeks after the accident. Both mother and child left the hospital in good condition.


Successful delivery after hepatic rupture in previous pre-eclamptic pregnancy. Rupture of the liver in the course of pre-eclamptic pregnancy is a rare but life-threatening event. Controversiality exists with regard to the treatment modality. A case is presented of a pre-eclamptic multiparous woman with liver rupture and intra-abdominal bleeding immediately after delivery of a stillborn infant, who was successfully treated by correction of intravascular volume with blood transfusions without surgical intervention. The index pregnancy was succeeded by an uneventful pregnancy and delivery. This is the first case report of a conservatively managed liver rupture with an uneventful course of the next pregnancy. Conservative treatment should consist of correction of hypovolemia and clotting disorders, while surgical approach should be reserved for patients who cannot be stabilized hemodynamically.


Fulminant subarachnoidal hemorrhage and coma subsequent to sudden-presenting hypertension. An unusual case of pre-eclampsia is presented, where a previously normotensive primigravida suddenly developed hypertension which evolved to subarachnoidal hemorrhage and coma within minutes. Delivery by cesarean section was followed by progressive and complete recovery in the next few days.


Possible relationship between coding recognition amino acid sequence motif or residue(s) and post-translational chemical modification of proteins. 1. The "code-sequence" of N-glycosylation site(s), the amino acids located around O-glycosylation site(s), the sequence motifs of several kinases, the sequence motifs of--sulfation, amidation, isoprenylation, myristoylation, palmitoylation and N-acetylation, Aspartic and Asparagine hydroxylation-site, gamma-carboxyglutamate domain, phosphopantetheine attachment site etc. are extensively listed, compared to those reported by "PROSITE" Computer Screen Center and discussed. 2. The structural aspects of protein-DNA recognition are quoted as discussion and conclusion.


Chemoprevention of cancer: protease inhibitors. 1. The defense of the organism against cancer by inhibitors of proteolytic enzymes which are able to block the metastasizing stage of the disease is reviewed. 2. The contemporary views on the possible mechanisms of the process of prevention on both molecular and cellular levels are presented.


An improved method for the purification of lignin peroxidases from Phanerochaete chrysosporium INA-12: properties of two major isoforms. 1. Phanerochaete chrysosporium INA-12 secretes several lignin peroxidase isoenzymes. This paper reports an improved procedure for the purification of the different isoforms compared to those previously described. 2. Lignin peroxidases are first concentrated and prefractionated on fast-flow ion-exchangers which avoid concentration by ultrafiltration and dialysis. 3. Further purification is achieved by hydrophobic interaction chromatography and anion-exchange FPLC. 4. Two major forms were purified to homogeneity. Kinetic measurements and protein characterization (isoelectric points, phosphate content) suggest that they are similar to those produced by P. chrysosporium BKM strain.


Nuclear glycoproteins of hamster liver and Kirkman-Robbins hepatoma recognized by concanavalin A. 1. Glycoproteins recognized by Concanavalin A (ConA) have been identified in nuclei and nuclear fractions differing in sensitivity to micrococcal nuclease digestion from hamster liver and Kirkman-Robbins hepatoma. 2. The major ConA binding proteins from hamster liver and Kirkman-Robbins hepatoma nuclei have molecular weights about 27,000 and 57,000, and 38,000 and 49,000, respectively. 3. A distinct distribution of glycoproteins between fractions differing in sensitivity to nuclease digestion has not been observed.


Function of intracellular phospholipase A2 in vectorial transport of apoproteins from ER to Golgi. 1. The cytosolic fraction required in in vitro reconstituted intracellular transport of mucus glycoprotein apopeptide (apomucin) was isolated and its potential as transport supporting factor assessed by the quantitation of the gastric apomucin transferred to Golgi. 2. The experiments with the fraction promoting transport and delivery of apomucin to Golgi revealed that the active protein has the property of phospholipase A2 (PLA2) which assists ER vesicles fusion with Golgi. 3. The ability of the 76 kDa PLA2 to hydrolyze phospholipids and to support transport and fusion of ER vesicles with Golgi was abolished by phosphorylation and regained following dephosphorylation. 4. The data provide evidence that 76 kDa intracellular PLA2 is responsible for the fusion of ER-transport vesicles with Golgi. The process of fusion is accomplished by generation of lysophospholipids in fusing membranes.


Analysis of the interaction uridin 5'-diphosphoglucuronic acid with intestinal bilirubin UDP-glucuronyltransferase. 1. Bilirubin UDP-glucuronyltransferase Michaelis-Menten kinetic parameters for UDP-glucuronic acid were studied using native and digitonin activated microsomes obtained from rat intestinal mucosa. 2. The intestinal enzyme showed a lower apparent Vmax compared with the hepatic enzyme in both native and activated microsomes; digitonin pretreatment enhanced Vmax 4 times in the former enzyme and 2 times in the latter. 3. The affinity of UDP-glucuronic acid for the intestinal enzyme was about 2 times lower than that for the hepatic enzyme and it was not substantially modified by detergent neither in the intestine nor in the liver. 4. The lipid analysis of intestinal and hepatic microsomes showed that the former present a higher content of cholesterol and a lower phosphatidylcholine/sphingomyelin ratio than the latter, accordingly the estimation of membrane fluidity using the fluorescence anisotropy of 1,6-diphenyl-1,3,5-hexatriene indicated that intestinal microsomes are more "rigid" than the hepatic ones. 5. These characteristics would provoke a restrictive milieu surrounding the enzyme that modifies its kinetic properties thus limiting its participation in the metabolism of bilirubin.


Bilirubin production and conjugation from newly formed heme in isolated rat hepatocytes. 1. Heme synthesis from delta-aminolevulinic acid (delta-ALA) in freshly isolated rat hepatocytes was maximal at 100 microM with a rate of approx. 7 nmol being synthesized per g wet weight cells. 2. Approximately 8% of synthesized heme was converted to bilirubin and 50% of the newly synthesized bilirubin was conjugated. 3. The ratio of di to monoconjugate was approx. 2.5. Incorporation of delta-ALA into bilirubin was increased by additional delta-ALA, heme and was also doubled in cells isolated from animals treated with CoCl2. 4. Bilirubin formation was inhibited approx. 90% by in vitro treatment with heme oxygenase inhibitors zinc and tin protoporphyrin.


Characterization of the proteins from Melanoplus bivittatus that bind juvenile hormone, and a refined EFDA photolabeling technique. 1. Juvenile hormone (JH) is specifically bound by a protein from hemolymph and fat body cytosol of the grasshopper, Melanoplus bivittatus. 2. This protein has a native molecular weight of 331,000 and subunits of 77,000. 3. Proteins that bind JH were covalently photolabeled with a JH analog, epoxyfarnesyl diazoacetate (EFDA). Samples were irradiated in spot plates and hydroxyapatite was used to separate bound from free [3H]EFDA. Differential solubilization was used to extract unlinked [3H]EFDA and solubilize [3H]EFDA linked to protein. 4. Hemolymph proteins of M(r) 479,000, 240,000 and 77,000 also bound [3H]EFDA. 5. Proteins that bound [3H]EFDA were not vitellogenins.


Characterization of Ca(2+)-ATPase of Setaria cervi (Nematoda: Filarioidea): effect of phenothiazines and anthelmintics. 1. A Mg2+ independent, Ca(2+)-ATPase requiring high concentrations of Ca2+ (5 mM) for the activation, equally distributed in cuticle-muscular-hypodermis, genital organs and gastrointestinal tissues and mainly localized in 10,000 g pellet fraction, was identified in Setaria cervi, a bovine filarial parasite. 2. Filarial enzyme showed Km value of 3.33 mM for ATP as computed from the double reciprocal Lineweaver-Burk plot. 3. The enzyme could be completely solubilized by sonication with about 4-fold increase in specific activity of the enzyme. 4. The enzyme showed about 2-fold activation by the calmodulin fractions isolated from S. cervi and rat brain homogenates. 5. The enzyme was highly sensitive to inhibition with some phenothiazine derivatives. Trifluoperazine was observed to be the most potent inhibitor followed by promethazine and chlorpromazine. 6. Some anthelmintics viz. diethycarbamazine and centperazine were found to be highly potent inhibitors of the enzyme, significant inhibition of filarial Ca(2+)-ATPase was also observed with levamisole and suramine. 7. Studies indicate Ca(2+)-ATPase of S. cervi as a potential chemotherapeutic target.


Efflux of polyamines from human lymphocytes and from L 1210 cells. 1. In human lymphocytes alkalinization of the cytoplasm with monensin or NH4Cl promotes release of polyamines. The effect of NH4Cl is abolished by EGTA and diltiazem. 2. Concanavalin A also promotes an increase of the efflux, counteracted again by EGTA and diltiazem. 3. By effect of TPA, polyamine efflux is decreased in the first 90 min, and later increased. The activation is partially prevented by H7 and by sphingosine. 4. In contrast with human lymphocytes, L 1210 cells release actively endogenous polyamines, but slowly radioactive polyamines. 5. Concanavalin A does not activate the latter process; A 23187 and NH4Cl on the contrary promote a much higher increase in the efflux rate than in normal lymphocytes. EGTA and diltiazem partially counteract the effect of NH4Cl on the release of radioactivity.


Purification and characterization of collagenolytic property of renal cathepsin L from arthritic rat. 1. This paper describes the purification and characterization of collagenolytic property of renal cathepsin L isolated from kidney of rats rendered adjuvant arthritis. The enzyme was isolated by acid extraction, ammonium sulfate fractionation, Sephadex gel filtration, CM-Sephadex chromatography and Sephacryl S-300 chromatography. 2. The enzyme preparation was found to be homogeneous by gel filtration and SDS-polyacrylamide gel electrophoresis. The molecular weight of the enzyme was estimated to be 29,000. 3. Incubation of rat tail tendon collagen with purified cathepsin L resulted a conversion of cross-linked beta-chain dimers into uncross-linked alpha-chain monomers. The pH optimum for collagen degradation by purified cathepsin L was found to be 3.5. This optimal pH is shifted to 4.5 when haemoglobin was used as a substrate for the enzyme. 4. Various activators and inhibitors were tested for their influence on the activity of cathepsin L. The purified enzyme showed a maximal activity in the presence of EDTA. Cysteine was also found to increase the activity of cathepsin L. This enzyme was strongly inhibited by iodoacetate, p-chloromercurobenzoate, mercuric chloride but not inhibited by pepstatin or PMSF. E-64 and leupeptin were also found to be strong inhibitors for cathepsin L. The degradation of rat tail tendon collagen by cathepsin L was completely inhibited by E-64. 5. The results presented in this investigation suggest that cathepsin L play a crucial role in the pathogenesis of adjuvant arthritis.


Superinduction of ornithine decarboxylase by halogenated ribofuranosylbenzimidazoles. 1. The effect of dichlororibofuranosylbenzimidazole (DiCl-RB), an inhibitor of hnRNA synthesis and casein kinase-2 activity, on ornithine decarboxylase (ODC) was investigated in a difluoromethylornithine (DFMO) resistant, ODC overproducing cell line. 2. In cells growing in the absence of DFMO, DiCl-RB provoked a marked, but transient increase in ODC activity and immunoreactive ODC content. 3. The ODC response to DiCl-RB was prevented by cycloheximide and was not due to stabilization of the enzyme. 4. The dibromo derivative analogue (DiBr-RB) exerted similar effects on ODC, but was effective at lower concentrations. 5. The halogenated ribofuranosylbenzimidazoles were ineffective in cells growing in the presence of DFMO and containing higher levels of ODC protein.


Metabolism of [14C]arachidonic acid-labeled lipids in quiescent and OAG-stimulated ascites tumor cells. 1. A rapid uptake and esterification of [14C]arachidonic acid during the first 4 hr of cultivation of ascites cells in serum-deprived medium was observed followed by a fast turnover of the fatty acid. 2. Labeling and turnover of esterified arachidonate in individual phospholipid classes was in the order: phosphatidylcholine (PC) greater than phosphatidylinositol (PI) much greater than phosphatidylinositol-4-phosphate (PIP) and -4,5-bisphosphate (PIP2) greater than phosphatidylethanolamine (PE) greater than PE-plasmalogens. 3. In cells stimulated with 1-oleoyl-2-acetyl-sn-glycerol a transient course of arachidonic acid incorporation into PC, PI, PIP and PIP2 was determined peaking 30 min after stimulation, indicating both esterification and release under these conditions. 4. The release of arachidonate was blocked by quinacrine which is a specific inhibitor of phospholipase A2.


Proteolytic processing sites producing the mature form of human cathepsin D. 1. The proteolytic processing sites of human lysosomal aspartic protease cathepsin D at which the intermediate single-chain form was converted into the mature two-chain form were determined. 2. The two chains were isolated by reversed-phase HPLC in order to investigate the cleavage sites of the enzyme. 3. Protein sequencing of the heavy chain, which was presumed to be derived from the C-terminal side in the single-chain enzyme, gave an N-terminal Leu 105. In addition, it revealed that there were also minor sequences, which commenced with Gly 106 and Gly 107. 4. A small C-terminal peptide was isolated from the light chain, which had been digested with two kinds of exogenous proteases. Sequence determination of this peptide, which was characterized as a nonapeptide by mass spectrometry, suggested that the C-terminus of the light chain was Ser 98. 5. These results indicate that a Ser 98-Ala 99 bond and an Ala 104-Leu 105 bond are cleaved to release 6 amino acid residues between the two chains.


Succinic semialdehyde dehydrogenase from mammalian brain: subunit analysis using polyclonal antiserum. 1. NAD(+)-dependent succinic semialdehyde dehydrogenase was purified to apparent homogeneity from rat brain and highly purified from human brain. 2. Molecular exclusion chromatography of the purified enzymes on Sephadex G-150 and G-200 revealed M(r) values of 203,000 and 191,000 for rat and human, respectively. 3. Electrophoresis on sodium dodecylsulfate polyacrylamide gels revealed a single subunit of M(r) 54,000 for rat and 58,000 for human. Isoelectric focusing of the purified rat enzyme yielded a pI of 6.1. 4. For both proteins, Km values for short-chain aldehydes acetaldehyde and propionaldehyde ranged from 0.33 to 2.5 mM; Km values for succinic semialdehyde were in the 2-4 microM range. 5. The subunit structure of both enzymes was investigated in brain extracts and purified preparations by immunoblotting, using a polyclonal rabbit antiserum against the purified rat brain enzyme. 6. For rat and human extracts, single bands were detected at M(r) 54,000 and 58,000, comparable to findings in the purified preparations. Immunoblotting analyses in other species (guinea pig, hamster, mouse and rabbit) revealed single subunits of M(r) 54,000-56,500.


Interaction of Zn2+ and Cu2+ ions with glyceraldehyde-3-phosphate dehydrogenase from bovine heart and rabbit muscle. 1. Binding of Zn2+ and Cu2+ ions to GAPDHs from bovine heart and rabbit muscle resulted in a partial loss of enzymatic activity of both enzymes, in a time and metal ion concentration dependent manner. Cu2+ ions caused a much larger decrease of the activity than Zn2+ ions. 2. Addition of NAD+ or EDTA to either enzyme resulted in a protective effect on GAPDH activity. A similar protective effect was observed following addition of 2-mercaptoethanol to the enzyme solution. 3. The association constant for GAPDH-Zn2+ complex, calculated from equilibrium dialysis data, was 0.9 x 10(4) M-1 for the bovine heart GAPDH and 1.3 x 10(4) M-1 for the rabbit muscle enzyme. The association constant for GAPDH-Cu2+ complex was the same for both enzymes, 11.3 x 10(4) M-1. 4. Equilibrium dialysis data also revealed that in either enzyme the specific sites, binding the metal ions, are identical or very similar, and independent from each other. They are situated in the most conserved part of the enzyme molecule. 5. Some zinc was found in GAPDH preparations from bovine heart. It is discussed if Zn2+ ions could have a kind of modulation effect on GAPDH activity.


The gene for the helix-loop-helix protein, Id, is specifically expressed in neural precursors. While mammalian neurogenesis has been characterized extensively, the molecules involved in regulating neural cell determination and differentiation remain ill-defined. There is accruing evidence that various members of the basic helix-loop-helix (bHLH) protein family critically regulate these biological processes in a number of tissues. Id, a negative regulator of bHLH proteins, was found to exhibit peak gene expression during mouse embryogenesis with a striking pattern in the central nervous system. Id transcripts were specifically localized to undifferentiated neural precursors of the ventricular zone and were not present in their differentiated derivatives. In addition, in the peripheral nervous system, dorsal root ganglia sensory precursors, known to be undifferentiated while dividing, also expressed Id mRNA. However, in the sympathetic nervous system and adrenal medulla, where differentiation and division occur simultaneously in precursors, Id was not expressed. Since Id transcript abundance inversely correlated with differentiation, this protein, similar to its Drosophila homolog, extramacrochaetae, may play a negative regulatory role in neural differentiation.


What are and what are not imaginal discs: reevaluation of some basic concepts (Insecta, Holometabola). Some general aspects of the concept of imaginal discs in the Holometabola are reevaluated. Their monolayer character and continuity with the surrounding epidermis are confirmed. Studies on the imaginal discs of the silkworm (Bombyx mori) and data from the literature show that the discs and their peripodial cells produce cuticle during larval life, as well as at metamorphosis. In B. mori it is demonstrated that adult and larval antennae are produced by the same cells or their progeny. The results also suggest that segments of the typically three-segmented larval antenna of Holometabola are not scape, pedicel, and one-segmented flagellum; at least segments 2 and 3 are of flagellar origin. Based on these and some additional facts it is argued that: (1) No larval organs are "replaced" at metamorphosis, but strict "sequential homology" is always maintained. (2) Imaginal discs are not undifferentiated structures destined to form the adult after larval breakdown, cannot be unambiguously defined, and do not represent qualitatively different epidermal structures. Classical imaginal discs (invaginated and present also in pre-final larval instars) arose several times independently and were not present in the larvae of ancestral Holometabola. (3) Since the disc cells are not undifferentiated and "embryonic" (if these words have a defined meaning at all), it is unreasonable to expect that the processes taking place in discs at metamorphosis would differ fundamentally from those occurring in other diploid metamorphosing epidermal cells.


Developmental sequence of expression of voltage-dependent currents in embryonic Xenopus laevis myocytes. Although the development of several of the voltage-dependent currents in embryonic amphibian myocytes has been described, the overall muscle electrical development, particularly the relative times of expression of different voltage-dependent currents, has not been addressed in a single study under one set of conditions. We have found that, in mesoderm isolated and cultured from neurula stage embryos, myocytes are identifiable before they express voltage-gated currents. These ionic currents are absent from all Xenopus mesodermal cells during the late gastrula/early neurula stages of embryonic development. At about the time of first somite segregation an inward rectifier K+ current is expressed in some myocytes, followed within 2 hr by a delayed rectifier K+ current. The density of both currents increases fourfold over the next 24 hr in culture. A Na+ current is not expressed in large numbers of myocytes until late in this culture period, at about the time that a slow Ca2+ current appears. Under our culture conditions the myocytes have a very low chloride conductance. A fast inactivating component to the outward K+ current is expressed in all myocytes by 24 hr in culture. In some experiments we dissociated embryos at later times and made recordings when all previously isolated myocytes expressed currents. In the late dissociations, most myocytes did not express currents, but developed them after a short period in culture. Because we have evidence that in vivo development is more closely approximated by the early dissociations, these results suggest that dissociation causes some degree of dedifferentiation.


Developmentally regulated expression of three slow isoforms of myosin heavy chain: diversity among the first fibers to form in avian muscle. At least three slow myosin heavy chain (MHC) isoforms were expressed in skeletal muscles of the developing chicken hindlimb, and differential expression of these slow MHC isoforms produced distinct fiber types from the outset of skeletal muscle myogenesis. Immunohistochemistry with isoform-specific monoclonal antibodies demonstrated differences in MHC content among the fibers of the dorsal and ventral premuscle masses and distinctions among fibers before splitting of the premuscle masses into individual muscles (Hamburger and Hamilton Stage 25). Immunoblot analyses by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of myosin extracted from the hindlimb demonstrated the presence throughout development of different mobility classes of MHCs with epitopes associated with slow MHC isoforms. Immunopeptide mapping showed that one of the MHCs expressed in the embryonic limb was the same slow MHC isoform, slow MHC1 (SMHC1), that is expressed in adult slow muscles. SMHC1 was expressed in the dorsal and ventral premuscle masses, embryonic, fetal, and some neonatal and adult hindlimb muscles. In the embryo and fetus SMHC1 was expressed in future fast, as well as future slow muscles, whereas in the adult only the slow muscles retained expression of SMHC1. Those embryonic muscles destined in the adult to contain slow fibers or mixed fast/slow fibers not only expressed SMHC1, but also an additional slow MHC not previously described, designated as slow MHC3 (SMHC3). Slow MHC3 was shown by immunopeptide mapping to contain a slow MHC epitope (reactive with mAb S58) and to be structurally similar to a MHC expressed in the atria of the adult chicken heart. SMHC3 was designated as a slow MHC isoform because (i) it was expressed only in those muscles destined to be of the slow type in the adult, (ii) it was expressed only in primary fibers of muscles that subsequently are of the slow type, and (iii) it had an epitope demonstrated to be present on other slow, but not fast, isoforms of avian MHC. This study demonstrates that a difference in phenotype between fibers is established very early in the chicken embryo and is based on the fiber type-specific expression of three slow MHC isoforms.


Position-specific expression of the annulin protein during grasshopper embryogenesis. Annulin, named for its annular expression in developing limb buds, is a approximately 100 kDa membrane-associated protein that is expressed in a complex and changing pattern during grasshopper embryogenesis. Its expression is dynamic along the developing midline and in the mesoderm, transient in neuroepithelial sheath cells around mitotic neuroblasts, and position-specific in circumferential stripes in each limb bud segment. Annulin expression begins along the midline of the embryo at the onset of gastrulation. Mesoderm cells express the protein as they migrate away from the midline as do new cells that come to lie at the midline. During neurogenesis, annulin expression disappears from many midline cells until only a specific subset of midline glial cells expresses high levels of the protein. Starting at the beginning of neurogenesis, sheath cells express annulin in correlation with the mitotic activity of the neuroblasts they surround.


Differential transcription of Pgk genes during spermatogenesis in the mouse. We have analyzed the occurrence of transcripts produced from the ubiquitously expressed, X-linked Pgk-1 gene and the testis-specific, autosomal Pgk-2 gene during spermatogenesis in the mouse. We found that tissue specificity, developmental specificity, and cell-type specificity of these mRNAs parallel that previously reported for the two protein isozymes of phosphoglycerate kinase (PGK) encoded by these two genes. This indicates that primary regulation of differential expression of the Pgk genes during spermatogenesis is exerted at the transcriptional level. We first detected Pgk-2 mRNA in preleptotene spermatocytes, indicating that transcription of Pgk-2 is initiated coincident with the onset of meiosis in male germ cells, and then continues to increase in later spermatocytes and postmeiotic round spermatids. This expression initiates prior to an initial decline in Pgk-1 transcript levels observed in pachytene spermatocytes, which apparently follows inactivation of the single X chromosome in spermatogenic cells. However, unlike cessation of Pgk-1 transcription from the inactivated X chromosome in female somatic cells, we show that inactivation of the Pgk-1 locus in spermatogenic cells is not followed by methylation of a key CpG dinucleotide in the promoter region. These results support the idea that specific expression of the Pgk-2 gene in meiotic and postmeiotic spermatogenic cells has evolved to compensate for reduced levels of Pgk-1 gene product caused by transient X-chromosome inactivation in these cells. They further suggest that reinitiation of transcription of the paternal Pgk-1 allele shortly after fertilization is facilitated by constitutive hypomethylation in the promoter region of this gene throughout spermatogenesis.


FGF-mediated aspects of skeletal muscle growth and differentiation are controlled by a high affinity receptor, FGFR1. Fibroblast growth factors (FGFs) and FGF receptors (FGFRs) play major roles in vertebrate embryogenesis, including control of skeletal muscle growth and differentiation. Understanding their roles requires delineating the specific FGF and FGFR isoforms involved. This study analyzes the FGFR transcripts found in a model mouse skeletal myoblast cell line (MM14) during growth and terminal differentiation. MM14 cells express transcripts for FGFR1 (flg) but not FGFR2 (bek). The predominate FGFR1 transcript contains three immunoglobulin (Ig)-like domains in the extracellular ligand binding region. Approximately one-fourth of the three Ig-like domain transcripts possess a 6-nt deletion between the first and second Ig-like domains which after translation would result in deletion of an Arg-Arg pair. Cloning of mouse genomic DNA surrounding the region of the FGFR1 6-nt deletion indicates that the deletion is derived by alternative splicing of FGFR1 transcripts. Transcripts containing two Ig-like domains account for less than 5% of total FGFR1 mRNA in MM14 cells. A survey of RNA from mouse tissues indicated that two Ig-like domain FGFR1 transcripts are rare in all tissues except in lung, in which the two Ig-like domain form accounts for roughly 70% of the lung FGFR1 mRNA. PCR RACE cloning studies disclosed 162 nt of additional FGFR1 5'-flanking RNA which was highly GC-rich. FGFR1 transcripts decline 8- to 10-fold during low serum, (-)FGF-mediated differentiation of MM14 cultures. The kinetics of the FGFR1 mRNA decline is similar to the previously described differentiation-dependent decrease in cell surface FGF receptors.


Mutant yolk proteins lead to female sterility in Drosophila. Specific mutations in the yolk protein genes, yp1 and yp2, of Drosophila melanogaster cause the yolk proteins (YPs) they encode to precipitate, ultimately resulting in female sterility. YPs of the yp1 mutant fs(1)1163 are secreted normally but then precipitate as globules and occasionally as crystalline fibers in the subbasement membrane space of the fat body (Butterworth et al., 1991, J. Cell Biol. 112, 727-737). The present ultrastructural and immunological studies of the fat body of the yp2 mutant fs(1)K313 show that YP also precipitates as globules in the same tissue compartment. The globules are also incapable of passing into the hemolymph but they are morphologically distinct from those of fs(1)1163. Similar analyses were performed on developing oocytes in wild type and both mutant strains. YP-containing aggregates, ultrastructurally similar to those in the fat body of each respective mutant, were found in the space between the plasmalemma and the vitelline membrane and embedded within the membrane itself. The evidence suggests that the precipitates interfere with the correct assembly of the eggshell membranes, leading to the sterile phenotype. Immunogold studies demonstrate that newly synthesized YPs in the normal and mutant strains share secretory vesicles with putative, vitelline membrane proteins and that the translocation of follicle cell YP is not through the membrane along the interfollicular spaces but directly through the plasmalemma facing the oocyte. Further the YP precipitates in the mutants permit visualization of the polarity of exocytosis of YP from the follicle cells.


Heterogeneity of Tau proteins during mouse brain development and differentiation of cultured neurons. Tau microtubule-associated proteins constitute a group of developmentally regulated neuronal proteins. Using the high-resolution two-dimensional polyacrylamide gel electrophoresis system, we have resolved more than 60 distinct Tau isoforms in the adult mouse brain. Tau protein heterogeneity increases drastically during the second week of brain development. In neuronal primary cell cultures, some of these developmental changes can be observed. The increase of Tau heterogeneity in culture is more limited and reaches a plateau after a period corresponding to the second week of development. Most, if not all, of the vast Tau heterogeneity can be attributed to intensive post-translational phosphorylation, which may affect the structure of the proteins.


Behavior of structurally divergent alpha-tubulin isotypes during Drosophila embryogenesis: evidence for post-translational regulation of isotype abundance. Two major alpha-tubulin isotypes are present during Drosophila embryogenesis: an evolutionarily divergent maternal isotype that is synthesized only in the ovary and deposited in the oocyte and a highly conserved constitutive isotype that is both maternally supplied and zygotically synthesized. A maternal isotype-specific antibody and a monoclonal antibody that recognizes both the maternal and constitutive isotypes were characterized and used to determine the distribution and abundance of alpha-tubulins during embryogenesis. Both isotypes are abundant and assemble into all classes of microtubules from the syncytial blastoderm stage until completion of germ band retraction. During subsequent development, however, the maternal isotype is retained only in the developing CNS, and later in a subset of connective fibers within the CNS. In contrast, total alpha-tubulin levels remain high in essentially all tissues throughout embryogenesis, indicating that most tissues selectively accumulate the constitutive isotype. To determine if selective accumulation of the constitutive isotype requires zygotic synthesis of this protein, mutant embryos that do not contain functional constitutive alpha-tubulin genes were examined. In these embryos, as in wild type, the maternal isotype decreases to background levels in tissues that retain high levels of the constitutive isotype. The constitutive isotype therefore appears to be more stable than the maternal isotype in most tissues. Differences in isotype stability may play an important role in determining the developmental pattern of isotype accumulation in Drosophila embryos.


Perturbations in morphogen gradients induce budding in hydra. Lateral grafting of small pieces of midregion tissue into different levels of the hydra body column was done to assess the influence of the host hypostome and basal disc (or, of the underlying morphogenetic gradients) in inducing secondary structures in the transplanted tissue; and also to identify the role, if any, of the induced secondary structures (or, perturbed morphogen gradients) on the pattern of the host. The same midpiece tissue differentiated to a basal disc when grafted near the host hypostome, and to a small hypostome with tentacles when grafted near the host basal disc. Chimeras with induced secondary basal discs showed a phenomenal increase in budding compared to the controls and to the chimeras having induced hypostomes. These results indicate a positive cross-reaction between both organizing regions during patterning in hydra.


The rapid transient decrease of sn-1,2-diacylglycerol in progesterone-stimulated Xenopus laevis oocytes is the result of an ethanol artifact. Full-grown Xenopus laevis oocytes resume meiosis (meiotic maturation) in response to progesterone stimulation. Three studies have shown that sn-1,2-diacylglycerol (DAG), the intracellular activator of protein kinase C, may be involved in this process (Wasserman et al., J. Exp. Zool. 255, 63-71, 1990; Varnold and Smith, Development 109, 597-604, 1990; Stith et al., J. Cell Physiol. 149, 252-259, 1991). Two of these studies (Varnold and Smith, 1990; Stith et al., 1991) found a rapid, but transient decrease in the levels of DAG of approximately 25 to 30% within 5 to 30 sec following the addition of progesterone to the oocytes. We have investigated this rapid decline in oocyte DAG. We also found a 20 to 34% decrease in DAG/oocyte within the first 5 to 40 sec following the addition of steroid to the culture medium. However, a similar rapid and transient decrease in oocyte DAG levels was also observed in response to ethanol. Ethanol is used as the vehicle to deliver progesterone to the oocyte culture medium. Therefore, the rapid transient decline in DAG appears to be an artifact of ethanol perturbing the production and/or turnover of DAG within the oocyte and not a physiological response of the oocyte to progesterone.


Ameiotic, a gene that controls meiotic chromosome and cytoskeletal behavior in maize. Microtubule organization during the novel cell division of ameiotic microsporocytes was examined using indirect immunofluorescence microscopy. A recessive mutation of the maize gene Ameiotic causes the replacement of meiosis I with a synchronized mitotic division (Palmer, R. G. (1971). Chromosoma 35, 233-246). All identifiable cytological features of this division, including chromosome behavior and microtubule organization, were typical of somatic cell division. Significantly, a cortical microtubule band was observed during prophase in ameiotic cells. In most somatic plant cells, a preprophase band of microtubules (PPB) predicts the cortical site where the future cell plate will join the sidewall. Similar structures, however, are absent in all meiotic and postmeiotic reproductive cells examined to date. These disruptions are consistent with a model where the wild-type Ameiotic gene encodes a product which acts during or before G2 and is necessary for initiating several independent meiotic processes, including both meiotic chromosome behavior and microtubule organization. The ameiotic mutation provides additional evidence that aspects of cytoskeletal organization unique to meiosis are genetically controlled. Finally, the presence of a PPB during the ameiotic division supports a model whereby multiple mechanisms are used to determine and maintain division plane polarity during normal meiosis.


The east gene of Drosophila melanogaster is expressed in the developing embryonic nervous system and is required for normal olfactory and gustatory responses of the adult. Drosophila melanogaster larvae and adults respond to a wide range of chemosensory stimuli. We describe the genetics and developmental expression of the east gene, mutations which result in adult-specific chemosensory defects. The original isolate of east is semidominant for the behavioral phenotype. Several mutations have been generated, some of which are recessive lethals and others that are viable alleles that show a recessive, adult-specific, chemosensory defect. No larval chemosensory defects were observed. The east gene is expressed in the neurogenic region at the time of neuroblast segregation and in cells in the peripheral and central nervous system. Our results suggest that east+ expression in the nervous system is required for a normal adult chemosensory response and both increases and decreases in levels of the gene product result in a mutant phenotype.