In vitro potentiation by lonidamine of the cytotoxic effect of adriamycin on primary and established breast cancer cell lines.

Lonidamine is a new potential chemotherapeutic agent, relatively non-toxic, that can positively modulate the efficacy of several antineoplastic drugs. We evaluated the response of two established human breast cancer cell lines (MCF-7 and BRC-230) and of 20 primary breast cancer cell lines to lonidamine, either alone or in combination with adriamycin, the drug most widely used in the management of breast cancer. Different schedules were tested by varying either concentration of the drugs (LND: 10-150 micrograms/ml; ADM: 0.10-0.15 micrograms/ml), or time of exposure (1-96 hours), or sequence of administration (ADM-->LND; LND-->ADM; ADM+LND). Our results indicate slight sensitivity of the cell lines to lonidamine when used alone, whereas an increase of efficacy was noted when lonidamine was added for at least 24 hours after a 4 hour exposure to adriamycin. Such efficacy was significantly greater than that expected from an additive effect between the two drugs. We conclude that lonidamine, when given according to an appropriate schedule, enhances, in vitro, the efficacy of adriamycin. A correct employment of lonidamine in the management of breast cancer might therefore potentiate the therapeutic effect of adriamycin.


A bioassay for antiestrogenic activity--potential utility in drug development and monitoring effective in vivo dosing.

Monitoring effective antiestrogenic activity of the triphenylethylenes in patients with breast cancer is usually determined by the duration of response. The pharmacokinetics of toremifene and tamoxifen have been shown to be highly variable but patient specific. In the present study, we developed a method to accurately assess the antiestrogenic activity of these agents using plasma specimens, cell culture, and cell cycle measurements. Plasma specimens (4-5mls) obtained from patients receiving toremifene (360mg/day for 5 days in a phase I trial) or tamoxifen (20mg/day) were extracted and reconstituted in tissue culture media (4-5mls), and growth inhibition was determined in estrogen responsive MCF-7 cells. Additionally, plasma specimens were quantified for toremifene or tamoxifen concentrations using HPLC. Growth inhibition of plasma specimens containing either toremifene or tamoxifen and their metabolites was also examined. Cell cycle measurements were determined following in vitro exposure with flow cytometric techniques. Our results show that a dose-response relationship exists between cell growth inhibition and cell cycle measurements for human plasma with added toremifene or tamoxifen, and also for human plasma specimens containing drug and its metabolites after treatment. Our antiestrogenic bioassay can address clinical research problems such as patient-specific pharmacokinetics, dosing compliance, and acquired antiestrogen resistance.


Monitoring the chemosensitizing effects of toremifene with flow cytometry in estrogen receptor negative multidrug resistant human breast cancer cells.

The clinical study of compounds that modulate multidrug resistance in cancer cells has been hindered by both the toxicities of these agents and the inability to monitor their effectiveness at a cellular level. The non-steroidal triphenylethylene toremifene is well tolerated clinically and can sensitize multidrug resistant cells to the effects of doxorubicin in vitro. The chemosensitizing properties of toremifene in estrogen receptor negative, multidrug resistant MDA-A1 human breast cancer cells were studied using flow cytometric analysis. Cell cycle kinetics of MDA-A1 cells were not significantly affected by treatment with either toremifene or doxorubicin alone, as the majority of cells remained in G0/G1. However, preincubation with toremifene for 70 hours followed by treatment with doxorubicin caused a marked shift of cells to G2, as cells appeared to be blocked in that phase of the cell cycle. This result was nearly identical to the effect of doxorubicin alone on doxorubicin-sensitive MDA-MB-231 breast cancer cells and can be interpreted as a "resensitization" by toremifene of MDA-A1 cells to doxorubicin. This chemosensitizing effect of toremifene was accompanied by an enhanced accumulation of doxorubicin in MDA-A1 cells (+110% after 70 hours pre-incubation with toremifene), and by a depression in protein kinase C activity in MDA-A1 cells that was maximal following 70 hours incubation with toremifene. Flow cytometry is a widely available technique that might be applied clinically to monitor at the cellular level the chemosensitizing effects of toremifene and other modulators of multidrug resistance.


A case-control study of breast cancer among Japanese women: with special reference to family history and reproductive and dietary factors.

To study the effects of family history and reproductive, anthropometric, and dietary factors on the risk of breast cancer among low risk populations, we conducted a hospital-based case-control study involving 908 patients with breast cancer and their matched controls, in Japan. A positive family history of breast cancer significantly increased the risk of breast cancer (odds ratio = 1.52, 95% confidence interval: 1.14-2.03). The risk further increased with increasing number of family members affected. Obesity, single marital status, fewer births, a late childbirth, and less consumption of green-yellow vegetables and dairy products were also associated with an increased risk of breast cancer. These associations were independent in multivariate analyses. There was no increase in risk associated with consumption of high fat foods. When analyzed by menopausal status, the association with family history of breast cancer, especially in the first degree of relatives, was more evident for premenopausal breast cancer. The associations with obesity and lower consumption of dairy products were more pronounced for postmenopausal breast cancer, while those with lower parity and single marital status were stronger for premenopausal breast cancer.


Expression of the pS2 gene in breast tissues assessed by pS2-mRNA analysis and pS2-protein radioimmunoassay.

The expression of the pS2 gene in breast tissues was assessed by measuring pS2-protein using a radioimmunoassay, and by determining pS2-mRNA using Northern blotting. There was a good correlation between the two measurements, indicating that expression of the pS2 gene in breast tissues may be assessed by either method. Since radioimmunoassay is technically easier and more efficient than Northern blotting, radioimmunoassay will be the method of choice in routine applications.


Type B lactic acidosis and metastatic breast cancer.

A case is presented of a patient with longstanding metastatic breast cancer whose condition suddenly deteriorated due to hypercalcemia and severe lactic acidosis which rapidly proved fatal. Postmortem examination showed no explanation for the lactic acidosis other than extensive metastatic disease. A review of the rare syndrome of malignancy-induced lactic acidosis is presented with particular emphasis on the 5 other cases reported in association with metastatic breast cancer. Theories of pathogenesis and management controversies are discussed.


Oral susceptibility of Aedes albopictus to dengue type 2 virus: a study of infection kinetics, using the polymerase chain reaction for viral detection.

Female Aedes albopictus mosquitoes, aged 1 week, were infected with DEN-2 dengue virus. The kinetics of infection in mosquito brain and mesenteron were monitored using DNA probes with polymerase chain reaction (PCR) amplification of target DNA sequences coding for DEN-2 virus envelope protein, compared with the standard immunofluorescence assay technique (IFA). Rates of virus detection in the mesenteron of orally infected mosquitoes rose to 38% by day 4 post-inoculation, then declined until day 8, followed by irregular peaks around days 11-14 and subsequently. In mosquito head squashes, virus was detected from day 4 onwards, reaching 38% positive by day 18. Salivary glands of all the same females were found to be positive for virus by day 8 onwards. Parenterally infected Ae.albopictus females were all positive for DEN-2 in the brain and salivary glands 8 days post-inoculation. In every case, results obtained with the PCR matched those from the IFA. Our DNA probe with PCR procedure can therefore be utilized as a sensitive and reliable method for studies of DEN-2 vectors.


Evaluation of neem, Azadirachta indica, with and without water management, for the control of culicine mosquito larvae in rice-fields.

Applications of neem, Azadirachta indica (Meliaceae), to rice-fields were evaluated with the dual objective of controlling the culicine mosquito vectors of Japanese encephalitis virus (JEV) and enhancing the grain yield. Since crude neem products deteriorate under improper storage conditions, a laboratory bioassay was developed to screen neem cake powder against mosquito larvae, Culex quinquefasciatus. Only samples of neem giving over 90% bioassay mortality were used in field trials. When good quality neem cake powder was applied at the dose of 500 kg/ha, either alone or coated over urea, there was a striking reduction in the abundance of late instar culicine larvae and pupae. Only fourteen pupae were obtained over a period of 13 weeks in neem cake powder treated plots, and four in those treated with neem coated urea, compared with 101 in control plots. Both treatments were significantly less than the control, but on par with one another. In another field trial, neem cake coated urea was tested at 500 and 250 kg neem/ha in combination with water management practices. No reduction in efficacy was noted at the lower dose. Larval abundance in plots under water management alone did not differ significantly from the controls, but was significantly reduced when water management was combined with neem products. Two stable formulations, 'Neemrich-I' (lipid rich) and 'Neemrich-II' (azadirachtin rich), also gave good suppression of immature culicines. All the treatments with neem also gave higher grain yield than the control.(ABSTRACT TRUNCATED AT 250 WORDS)


Biting and resting behaviour of anophelines in western Venezuela and implications for control of malaria transmission.

A study was conducted in three villages in western Venezuela between February 1988 and October 1989 to determine the biting and resting activity of anophelines in relation to human habits, rainfall and fenitrothion spraying of houses. Mosquitoes were collected landing on humans inside and outside experimental huts throughout the night. Only three mosquitoes were found resting in the huts in the morning, but 2470 were collected resting on vegetation in the early morning. The collections yielded eleven species of anopheline, the most abundant being Anopheles nuneztovari, comprising over 75% of the total anophelines collected, followed by An.triannulatus, An.albitarsis s.l. and An.oswaldoi. The four most abundant species showed different diel patterns of biting. For An.nuneztovari the peak of activity was close to midnight indoors and outdoors, for An.triannulatus between 19.00 and 20.00 hours outdoors, for An.albitarsis mainly before midnight indoors and outdoors and for An.oswaldoi outdoors at 19.00 hours, with an additional smaller peak indoors at midnight. Most of the human population use bednets, go to bed before 22.00 hours and are therefore most exposed to mosquitoes that bite outdoors early in the night. Fenitrothion house-spraying failed to prevent large mosquito populations developing in the wet season, presumably because of their exophilic resting habits. The possible advantages of impregnation of existing bednets with pyrethroids, and provision of nets for people who do not have them, are discussed.


Population genetic confirmation of species status of the malaria vectors Anopheles willmori and An. pseudowillmori in Thailand and chromosome phylogeny of the Maculatus group of mosquitoes.

Among the Anopheles (Cellia) maculatus group of Oriental mosquitoes, positive assortative mating occurs within sympatric and synchronous populations of An.willmori and An.pseudowillmori in the presence of populations of An.maculatus and An.sawadwongporni, judged from the occurrence of inversion homozygotes and the absence of any heterozygotes in their distinctive, polytene chromosomes. Genotypic frequencies for enzyme electromorphs give additional evidence for the species status of An.pseudowillmori and a practical means of identification in field studies of malaria vectors. Autosomal rearrangements are referred to those of An.stephensi which is unique for 4y. An.willmori differs by a single inversion, 4x; An.pseudowillmori by three Arm 2 inversions; and An.dispar by one Arm 2 inversion and 4x. Fixed autosomal rearrangements in the Maculatus group are summarized and their phylogenetic distribution suggests some unknown, intrinsic mechanism by which genome structure is disrupted in association with speciation events. This could be relevant to the potential genetic manipulation of malaria vectors.


Insecticide resistance gene frequencies in Anopheles sacharovi populations of the Cukurova plain, Adana Province, Turkey.

In Turkey, the mosquito Anopheles sacharovi has been under field selection pressure sequentially with DDT, dieldrin, malathion and pirimiphosmethyl over a period of 30 years for the purpose of malaria control. In 1984, the field population of An.sacharovi in the malarious Cukurova plain of Adana Province contained an altered acetylcholinesterase-based resistance gene giving broad spectrum resistance against organophosphorus and carbamate insecticides. The cross-resistance spectrum from this mechanism conferred resistance to malathion but not to the organophosphorus insecticide pirimiphos-methyl. Over the 6 years that pirimiphos-methyl has been applied for malaria vector control in this area, the frequency of the altered acetylcholinesterase resistance gene has declined, although in 1989 and 1990 it was still present at measurable frequencies in An.sacharovi from Cukurova. In addition to the acetylcholinesterase resistance mechanism there is evidence of an increased level of glutathione S-transferase in some of the An.sacharovi populations tested. This is known to be correlated with DDT resistance in other anophelines. In Turkish An.sacharovi, DDT resistance and elevated glutathione S-transferase occur in the same populations at similar frequencies. The continued prevalence of resistance to DDT and dieldrin, long after the 1971 cessation of DDT spraying for malaria control in Turkey, suggests that the DDT resistance gene has insufficient reduced fitness associated with it to have been lost from the field population during the past two decades. The implications of the slow decline in resistance gene frequencies in this field population are discussed in relation to mathematical models for managing resistance.


Activation of three species of tsetse (Glossina spp.) in response to host derived stimuli.

Recordings were made of the activation of hungry Glossina morsitans morsitans Westwood, G. pallidipes Austen, and G. austeni Newstead in response to odours from ox breath and ox urine, and a moving visual stimulus, in a wind tunnel. The spontaneous activity of G.m.morsitans was very low (less than 4% of males and 2% of females active per min during control periods). That of G.austeni and G.pallidipes was in the region of 20% except for G.pallidipes females when in excess of 40% were active during control periods. Addition of ox urine odours to the airstream had no effect on activity in any of the species investigated but addition of ox breath odours to the airstream significantly increased activity of G.pallidipes and of G.m.morsitans, although for the latter only approximately 12% of flies were active. For G.austeni the addition of ox breath odours resulted in a significant increase in activity of females but not of males. The moving visual stimulus resulted in a significant increase in the activity of both sexes of G.austeni and G.m.morsitans but no change in the activity of G.pallidipes. The low level of spontaneous activity and the low response to ox breath odours in a strain of G.m.morsitans maintained in the laboratory since 1969 was compared with a new colony of this species which originated from puparia collected in Zimbabwe in 1991. No differences in either spontaneous activity or the response to ox breath odour was recorded, but females from the new colony were significantly more responsive to a moving visual stimulus.(ABSTRACT TRUNCATED AT 250 WORDS)


Abiotic factors influencing embryonic development, egg hatching, and larval orientation in the reindeer warble fly, Hypoderma tarandi.

Wild-caught, tethered females of the reindeer warble fly, Hypoderma tarandi (L.) (= Oedemagena tarandi (L.)), (Diptera, Oestridae) were stimulated to oviposit on hairs of a reindeer hide. Newly laid eggs incubated at constant temperatures and relative humidities hatched within 3 days to 2 weeks, depending on the experimental conditions. Over a range of 7-40 degrees C, hatching only occurred between 20 and 37 degrees C. Eggs held at 100% relative humidity had lower hatchability and longer time to hatch relative to eggs held at 77% relative humidity. The average number of degree-days for hatching was 50.35. Between 20 and 33 degrees C there was a temperature-dependent linear trend in developmental rate, and the proportion of eggs hatching was highest, and least variable, at the mid-temperature ranges. The temperature range found in the natural host micro-habitat where H. tarandi commonly affix their eggs (close to the skin at the base of a host hair) was consistent with the experimental temperature treatments that produced the highest hatching rate. Newly emerged larvae displayed positive thermotaxis, while showing no phototaxic or geotaxic behaviour. Results indicate that constraints of the host environment, coupled with temperature-dependent hatching success, may impose a selective pressure on oviposition behaviour.


Tick infection rates with Borrelia: Ixodes ricinus versus Haemaphysalis concinna and Dermacentor reticulatus in two locations in eastern Germany.

Unfed nymphal Ixodes ricinus, Haemaphysalis concinna, and adult Dermacentor reticulatus were collected in two locations of Saxony in July and September 1991 by flagging. In July, the abundance of nymphal I. ricinus was about 2-3 times higher than that of nymphal H. concinna, a time of the year when nymphs of both species are reported to have a seasonal peak of activity. No D. reticulatus were flagged concurrently. In September, host-seeking activity of nymphal I. ricinus was again quite high as was that of adult D. reticulatus but only low numbers of nymphal H. concinna were collected. The flagged ticks were individually examined for Borrelia by an indirect immunofluorescence assay (I. ricinus: n = 414; H. concinna: n = 96; D. reticulatus: n = 116). The prevalence of Borrelia (probably B. burgdorferi) in I. ricinus varied from 12.1% to 21.0%. No borreliae were found in H. concinna. Of the examined D. reticulatus from one site (n = 97) 11.3% contained either B. burgdorferi or a related Borrelia. This may be the first finding of Borrelia in an Eurasian Dermacentor species.


Distribution of breeding and control of the filariasis vector Aedes samoanus in leaf axils of Pandanus in Samoa.

Water in leaf axils of the screwpine Pandanus was sampled for mosquito immature stages at seventy villages in Upolu, fifty-five in Savai'i and three in Manono, the main islands of Samoa. Ten plants in every patch of Pandanus plantation were sampled at each village. Among 23,049 mosquito larvae collected from Upolu, 77% were the filariasis vector Aedes (Finlaya) samoanus, 17.7% were Ae. (Fin.) oceanicus and 5.3% were Ae. (Fin.) tutuilae. Out of 6981 larvae taken in Savai'i, 23.2% were Ae. samoanus, 67.6% Ae. oceanicus and 9.2% Ae.tutuilae. When larval counts per plant were analysed for each district, Ae. samoanus was found to predominate in Pandanus in Upolu and Ae. oceanicus in Savai'i. However, the adult density of Ae.samoanus was higher in Savai'i and this was attributed to the large areas of forests with Freycinetia for Ae.samoanus breeding. In Pandanus in Savai'i the number of Ae.samoanus was negligible. In Upolu, with more urbanization and larger plantations, there was greater breeding of Ae.samoanus in Pandanus. Two control trials were conducted against Ae.samoanus larvae in Pandanus, one using a sand culture of the parasitic nematode Romanomermis culicivorax and the other with temephos, an organophosphate insecticide. While R. culicivorax did not adapt to the leaf axil habitat, all plants were without larvae for 5 weeks after treatment with temephos.


Seasonal abundance and biting behaviour of Anopheles punctulatus and An. koliensis in Malaita Province, Solomon Islands, and a trial of permethrin impregnated bednets against malaria transmission.

Seasonal abundance of the malaria vectors Anopheles punctulatus Dönitz and An.koliensis Owen in Bilimanu, an isolated inland village with forty-two houses in Malaita Province of the Solomon Islands, was monitored over 28 months by means of all-night landing/biting catches at one site during June 1985 to September 1987. Totals of 1250 An.punctulatus and 141 An.koliensis were collected, the latter being the largest number of this species ever caught at any locality in the Solomons. Bednets impregnated with permethrin 0.5 g/m2 were introduced in December 1986 to be used at night by all 190 villagers for protection against malaria vectors. Bioassay tests with An.punctulatus blood-fed females exposed under nets for 10 min resulted in 100% mortality up to 50 weeks post-impregnation. For An.punctulatus, the main vector species, the mean catch (indoors + outdoors) per man hour was 2.9 (range 0.7-13.2) before a cyclone on 19 May 1986, and, 0.66 (0.2-2.7) after the cyclone. The vector survival rates were usually high before the cyclone, but erratically lower thereafter for An.punctulatus. An.koliensis disappeared after the cyclone. Both An.punctulatus and An.koliensis consistently showed higher rates of biting man indoors than outdoors and their diel biting cycle showed a peak around midnight. Outdoors, the parous proportion of An.punctulatus was twice the nulliparous, and nearly so indoors. Following intervention with permethrin-treated bednets, the mean catch of An.punctulatus fell to 0.35 per man-hour (monthly range 0-1.5). The Plasmodium falciparum malaria infection rate reduced from 10% pre-intervention to zero in September 1987, 9 months after intervention, and then rose again.(ABSTRACT TRUNCATED AT 250 WORDS)


DNA probes for the identification of members of the Simulium damnosum complex (Diptera: Simuliidae).

Genomic libraries in plasmid have been constructed from various sibling species of blackflies of the Simulium damnosum complex from West Africa. Three cloned repetitive sequences, which show variation in copy number between sibling species, have been isolated. These clones can be used as probes for the dot-blot identification of larvae, pupae or adults into the three main West African subcomplexes, i.e. damnosum, squamosum and sanctipauli subcomplexes. The sequences also show some intraspecific variation in copy number.


Identification of phlebotomine sandfly bloodmeals from Baringo District, Kenya, by direct enzyme-linked immunosorbent assay (ELISA).

Direct enzyme-linked immunosorbent assay (ELISA) was used to identify the sources of bloodmeals in phlebotomine sandflies from Baringo District, Rift Valley Province, Kenya. Some bloodmeals had been stored for over 4 years before being analysed. Among 356 sandflies identified, 62.9% were Phlebotomus martini, 14.8% Sergentomyia antennatus, 10% S.schwetzi, 6% S.clydei, 1.9% S.adleri, 1.6% P.duboscqi, 1.4% S.africanus and 0.8% S.bedfordi. Out of 224 P.martini bloodmeals, host source was identified for 69. The order of host preference for P.martini was: goat 28.5%, rabbit 22.7%, human 8.9% and others 8.9%. Evidence of mixed feeding was shown by four species comprising sixteen specimens, twelve of which were P.martini. The most effective methods for trapping bloodfed P. martini were sticky paper traps in termite hills, followed by light-traps. Of the 224 P.martini trapped, 58.9% were collected with traps in termite hills, and 22.7% with light traps. Roles of the three most popular hosts for P.martini should be investigated to ascertain whether they act as reservoirs in the transmission of Leishmania donovani causing visceral leishmaniasis in Kenya.


Chigger mites of the genus Leptotrombidium: key to species and their distribution in China.

Chigger mites of the genus Leptotrombidium (Acari: Trombiculidae) transmit scrub typhus, caused by Rickettsia tsutsugamushi (= R.orientalis) in South-East Asia. In China, eighty-two species of Leptotrombidium have been recorded; these are listed with the names of Provinces where they were found. Five species, L.deliense, L.insularae, L.kaohuense, L.rubellum and L.scutellare, have been implicated as Chinese vectors of scrub typhus. A brief key is given to the larvae of all but three of the Leptotrombidium mites known in China.


Posterior vitreous detachment.

Posterior vitreous detachment is an expected consequence of aging, but it can also be the initiating cause of a retinal detachment. To understand the mechanism of posterior vitreous detachment and its sequelae, it is necessary to appreciate the anatomy of the vitreous, its development, and the pathogenesis of vitreous degeneration. This paper is a discussion of these considerations, the types of complications that may result from vitreous detachment, the proper examination of patients who present with the symptoms of vitreous detachment, and appropriate patient management.


Clinicolegal aspects of vitreous and retinal detachment.

Failure to diagnose retinal detachment is an important cause of professional liability claims involving optometrists. To comply with medical standards, dilation of the pupil and examination of the retinal periphery are required, both for patients who are at risk for retinal detachment and for patients who are symptomatic. Important risk factors include a history of previous detachment, myopia, lattice retinal degeneration, aphakia and pseudophakia, YAG capsulotomy, treatment of primary open-angle glaucoma with miotic drugs, blunt ocular trauma, and the proliferative retinopathies. Patients with the symptoms of acute posterior vitreous detachment must receive a dilated fundus examination, evaluation of the retinal periphery, and assessment of the vitreous. If examination results are negative, the patient should be warned of the symptoms of retinal detachment and scheduled for reevaluation in 4 to 8 weeks. Examination findings, warnings, recall appointments, and referral for treatment should be documented in the patient's record of care.


Scleral depression.

Indirect ophthalmoscopic examination of the peripheral retina is greatly enhanced through the use of scleral indentation. This procedure enables the examiner to better diagnose and manage peripheral retinal anomalies. In this study, the anatomical landmarks and structures relevant to examination of the peripheral fundus are reviewed, along with the various types of instrumentation used in performing scleral indentation. The mechanical techniques used in scleral indentation are discussed as are many of the common errors committed by those learning to perform this procedure. Scleral indentation can be especially valuable in determining the exact nature of lesions noted in the retinal periphery. Case studies in which this procedure was vital to reaching an accurate diagnosis have been included.


Retinal detachment.

Retinal detachment is a separation of the neurosensory retina from the retinal pigment epithelium. Most retinal detachments are rhegmatogenous, and identification of risk factors and predisposing lesions are important aspects of patient management. Retinal detachment is relatively rare, but can pose a significant threat to vision if there is macular involvement. Prompt diagnosis combined with patient education and appropriate intervention often can avert irrevocable visual impairment. This paper presents an overview of the categories of retinal detachment, discusses the pathogenesis of the various types of detachment, and provides recommendations for primary care of patients with predisposing factors and high-risk characteristics.


Lattice degeneration of the retina and retinal detachment.

Lattice retinal degeneration is considered the most significant peripheral retinal disorder potentially predisposing to retinal breaks and retinal detachment. Lattice degeneration affects the vitreous and inner retinal layers with secondary changes as deep as the retinal pigment epithelium and perhaps the choriocapillaris. Variations in clinical appearance are the rule; geographically, lattice lesions favor the vertical meridians between the equator and the ora serrata. Lattice degeneration begins early in life and has been reported in sequential generations of the same family. Along with its customary bilateral occurrence, lattice shares other characteristics of a dystrophy. The association between the vitreous and retina in lattice lesions may be responsible for the majority of lattice-induced retinal detachments. The tumultuous event of posterior vitreous separation in the presence of abnormally strong vitreoretinal adherence is the trigger for a retinal tear that, in turn, may lead to retinal detachment. Although retinal holes in young patients with lattice degeneration may play a role in the evolution of retinal detachment, the clinical course of lattice degeneration seems to be one of dormancy rather than of progressive change. This discussion outlines the pathophysiology of lattice retinal degeneration and the relationship of pathophysiology to clinical presentation. The epidemiology of lattice degeneration is summarized, as are the possible precursors to retinal detachment. A clinical characterization of the natural history of lattice degeneration is offered, and interventions for complications are described. To conclude, management strategies from a primary-care standpoint are reviewed.


Retinal dialysis.

Whether idiopathic or the result of ocular trauma, retinal dialysis is one of the least understood causes of retinal detachment. Dialyses are a common cause of retinal detachment and are the leading cause of traumatic retinal detachment in children and young adults. A dialysis is a disinsertion of the retina found at the ora serrata, generally involving less than 3 clock hours. Because a patient may be asymptomatic, a dialysis may be missed during ophthalmoscopic examination of the ocular fundus unless scleral indentation is used. Anatomical reattachment of detachments from retinal dialysis is highly successful for dialyses of both traumatic and idiopathic origin, with good visual results obtained postoperatively.


Methods of monitoring menstrual function in field studies: efficacy of methods.

Efficacy of methods for monitoring female reproductive potential under field study conditions was evaluated. Women (n = 10) were recruited to participate for two menstrual cycles on the bases, in part, of not seeking fertility assistance, working full-time but not in the medical field, and having less than one year of college education. Luteinizing hormone (LH), estrone-3-glucuronide, and pregnanediol-3-glucuronide were measured in daily morning urine and normalized to creatinine concentrations. These urinary measures were parallel to serum LH, estradiol, and progesterone profiles. Based on these urinary measures, 6 of 19 cycles were judged to be atypical. Transvaginal ultrasonography provided insights into ovarian activity during the atypical cycles. Of 13 LH surges detected by radioimmunoassay, 7 were not detected by a semiquantitative dipstick (OvuSTICK), perhaps due to that method's sensitivity to loss of LH immunoactivity caused by sample freezing. While intervals from salivary and vaginal mucous electrical resistance signals to the LH surge during typical cycles were similar to those reported previously, they were not predictive of ovulatory status during atypical cycles. Fifty-three percent of the cycles were misclassified on the basis of the basal body temperature rise. Cervical mucous color, amount, and consistency were not predictive of ovulation under these study conditions. The results from these 19 menstrual cycles provide information about the efficacy of various methods for characterizing menstrual function under field study conditions. In this regard, urinary endocrine measures are the most informative or practical.


Methods of monitoring menstrual function in field studies: attitudes of working women.

This study was designed to determine the attitudes and compliance of working women toward methods being evaluated for use in the assessment of the effects of toxicants on reproductive potential. Women such as the highly motivated fertility patients and nurses, who are typically familiar with the methods and procedures of fertility assessment and the value of medical research, have been used to validate such methods in a clinical setting. However, the attitudes of a general working female population toward these methods are unknown. Nine participants were selected on the bases, in part, of not seeking fertility assistance, working full-time but not in the medical field, and having less than one year of college education. Attitudes were also evaluated for 193 non-participating women to whom the procedures had been verbally described. Participants measured basal body temperature and salivary and vaginal mucous electrical resistance, evaluated cervical mucus manually (CME), and collected the first morning urine for two menstrual cycles. Blood, saliva, and transvaginal ultrasonograms (US) were obtained at a fertility clinic 6 to 9 days per cycle. Participants brought urine to the laboratory every 3 days. All participants performed all methods. Participants were paid $400; nonparticipants were not compensated. Only 3% of the respondents objected to the proposed methods: principally to CME, US, and giving blood samples. No respondent perceived the study as unimportant.(ABSTRACT TRUNCATED AT 250 WORDS)


Reproductive toxicity of cyclophosphamide in the C57BL/6N mouse: 1. Effects on ovarian structure and function.

The antineoplastic alkylating agent, cyclophosphamide (CPA) is known to impair normal female reproductive function. We have examined the time- and dose-dependent effects of CPA on the ovary, specifically, its impact on follicle numbers, ovarian morphometrics, and estradiol (E2) production. Female C57BL/6N mice were treated ip with CPA in normal saline at doses of 0, 75, 200, or 500 mg/kg. Ovaries were removed 1 to 14 days following treatment and serial sections were prepared. Differential follicle counts revealed that primordial follicles were most sensitive to CPA (ED50 = 122 mg/kg), followed by antral and growing follicles. Primordial follicles were affected by all doses of CPA and were completely destroyed by 3 days in the 500 mg/kg dose group. The greatest reduction in antral follicles was to 49% and 7% of controls by CPA doses of 200 and 500 mg/kg, respectively. Plasma E2 concentrations correlated best with antral follicle numbers (r2 = 0.94) and antral follicle volume (r2 = 0.88). Growing follicles were least sensitive to CPA and only decreased at 7 and 14 days. Although atretic changes were observed in growing follicles after treatment with CPA, these follicles recovered and progressed into apparently functional antral follicles (that is, they produced E2). Total ovarian volume was significantly reduced (30 to 40%) in the high-dose group on day 1, and remained depressed throughout the experiment. Examination of ovarian morphometrics indicated that this volume loss represented specific temporal changes in corpora lutea (CL), interstitial tissue, growing follicles, and antral follicles. At 1 and 3 days after treatment, the major loss in ovarian volume was due to a reduction in antral follicle and interstitial tissue volumes, while at 7 days the majority of volume loss was accounted for by the absence of CL. It is not known if CL are directly affected by CPA at the early time points, but their absence at 7 and 14 days is probably due to earlier destruction of antral follicles. These results demonstrate that CPA-induced ovarian toxicity is exhibited as temporal changes in both structural and functional features of the ovary, particularly in destruction of primordial and antral follicles and depressed E2 production. Information of this type also gives insight into ovarian response to chemical disruption of folliculogenesis and its recovery process.


Reproductive toxicity of cyclophosphamide in the C57BL/6N mouse: 2. Effects on uterine structure and function.

Cyclophosphamide-induced uterine weight loss was evaluated to determine whether it was a function of primary toxicity to the uterus or a secondary response to ovarian toxicity, that is, antral follicle destruction. C57BL/6N mice treated with cyclophosphamide exhibited a reduction in uterine weight concurrent with a decrease in plasma estradiol (E2) concentrations, thereby indicating toxicity to the ovary. However, when E2 concentrations recovered, uterine weight still remained depressed, suggesting that cyclophosphamide also impaired uterine function. Further investigation revealed that cyclophosphamide altered the normal uterotropic response to E2, significantly diminishing the uterine weight gain associated with E2 treatment. We conclude that effects of cyclophosphamide on the uterus involve two components: 1) decreased uterine weight in response to decreased plasma E2 resulting from ovarian toxicity, and 2) an altered response to E2 due to direct uterine toxicity.


Effects of methoxychlor treatment of pregnant mice on female offspring of the treated and subsequent pregnancies.

This study was designed to assess whether exposure to the estrogenic pesticide methoxychlor (MXC) during pregnancy would affect reproductive parameters not only in female offspring exposed prenatally, but also in those of a subsequent litter. Mice were exposed via oral gavage to 7.5, 5.0, or 2.5 mg technical grade MXC (50%) or 0.025 mg estradiol-17 beta (E-17 beta) from days 6 to 15 of pregnancy. Following delivery, female offspring (F1a) were cross-fostered and sacrificed at 8 weeks of age. Mothers exposed during their first pregnancy were allowed to mate again and their second set of offspring (F1b) were similarly evaluated to detect any latent effects from the initial exposure. Mice exposed to 7.5 mg MXC were unable to carry their litters to term. Results revealed a significant increase in the length of gestation of mice exposed to both E-17 beta and 5.0 mg MXC. A larger percentage of atretic follicles appeared in the ovaries of F1a females exposed prenatally to 5.0 mg MXC when compared to controls. Females from the F1b litter displayed a significant advance in time of vaginal opening, an apparent residual effect of MXC from a mother exposed during a previous pregnancy.


Effects of antiestrogens on adult and neonatal mouse reproductive organs.

Estrogenic potencies of various antiestrogens, including keoxifene (Kx) and trifluoperazine (Tfp), on reproductive tracts of ovariectomized adult mice, and effects of neonatal Kx and Tfp on reproductive organs were studied in C57BL/Tw mice. In adult ovariectomized mice, weight, DNA, and protein of the uterus and vagina were increased by 3 daily injections of 100 micrograms clomiphene, tamoxifen (Tx), and nafoxidine, and of 1 microgram estradiol-17 beta (E), but not by Kx. Antiestrogenic potency of Kx was studied in adult mice given injections of E. Kx significantly suppressed the E-induced increase in weight, DNA, and protein in the uterus and vagina. Tfp (20 micrograms), known as a tranquilizer and an antiestrogen, had no estrogenic effect on either organ. Male and female mice given 5 daily injections of Kx or Tfp from the day of birth were examined at 30, 40, and 60 days of age. Weights of testis, epididymis, and seminal vesicle in neonatally Kx-treated mice were significantly lower than in controls at 30 and 40 days. Spermatozoa were not formed in the seminiferous tubules of Kx-treated mice, although spermatogenesis occurred at 60 days. In neonatally Kx-treated females, weight of the uterus at 60 days and of the vagina at 40 and 60 days was significantly lower than in controls. Corpora lutea were absent from the ovaries of Kx-treated females. In neonatally Tfp-treated mice of both sexes at all ages examined, no differences were found in organ weights or histology, other than lower spermatogenic indices at 40 and 60 days of age.(ABSTRACT TRUNCATED AT 250 WORDS)


A 40-week male fertility study of the anticancer agent sulofenur (LY186641) administered orally to rats.

Sulofenur was evaluated for fertility effects on rats. Five-week old male rats (20/group) received 0, 5, 30, or 60 mg Sulofenur/kg/day for 14 weeks. Fertility was evaluated five times. Treated males were mated with untreated females at 10 weeks. Half the males from each group were necropsied after the 14-week treatment period and the remainder were mated four additional times during a 26-week posttreatment period. Following each mating, females were killed on gestation-day 20 and examined for evidence of pregnancy. Six weeks after mating trial 5, the remaining males were necropsied. Testes and epididymides were collected, weighed, and examined microscopically. All rats mated in each mating trial, and all control and low-dose animals were fertile. A significant reduction in fertility occurred in the middle- and high-dose males. This was consistent with testicular hypospermatogenesis in these groups. Fertility recovered in the high-dose group following cessation of treatment, but remained reduced in the middle-dose group. Preimplantation and postimplantation loss in mating trial 1 were higher in the middle- and high-dose groups. Abnormal fetuses were present in the high-dose group in mating trial 3, but not in mating trials 4 or 5. In conclusion, male rats given doses of 30 and 60 mg/kg/day of Sulofenur showed hypospermatogenesis and decreased fertility. Spermatogenesis and fertility recovered in the high-dose group. A dose of 5 mg/kg/day did not produce any effect on testes or fertility.


Relationship between fetal weight and litter size in rats: application to reproductive toxicology studies.

The inverse relationship between mammalian fetal weight and litter size has been discussed by many authors, but their opinions reveal no agreement at all. As in toxicity studies of reproduction, both parameters must be correctly evaluated. We investigated the existence of such a relationship in 2466 fetuses from 203 litters of Sprague-Dawley CD control rats. The frequency distribution of fetal weights had a normal adjustment. From the mean weight of fetuses in each litter, the mean fetal weights in each litter size and correlation coefficient were calculated and the regression line was plotted; the correlation coefficient (r = 0.677) was highly significant (P = 0.002), which made evident that there was an inverse relationship between fetal weight and litter size. If fetal weight/litter size inverse relationship is not taken into account when toxicity on the fetal weight is analyzed, wrong conclusions may be reached if the test substance reduces the litter size, provoking embryofoetal mortality. The iatrogenic decrement in fetal weight can be masked by an increment due to the litter size reduction. We suggest that in all three segments of reproductive toxicity studies, litter size must be considered as a covariate to the effect of the test substance on the fetal weight, in order to perform a correct analysis of covariance (ANCOVA), in addition to the dose factor commonly used in common ANOVA.


From cisplatin to artificial nucleases--the role of metal ion-nucleic acid interactions in biology.

Metal ions and metal coordination compounds bind to nucleic acids in a variety of ways, ranging from weak electrostatic interactions via hydrogen bonding and/or van der Waals forces to strong covalent binding. Metal ions naturally take part in the formation and the degradation of nucleic acids, and the propensity of certain metal coordination compounds to bind to nucleic acids, notably DNA, is exploited in cancer chemotherapy. Moreover, metal compounds have a wide potential as chemical probes for nucleic acid structures and as tools for nucleic acid processing.


Aluminum: a pH-dependent inhibitor of NADP-isocitrate dehydrogenase from porcine heart.

Aluminum showed a pH-dependent inhibitory effect on NADP-isocitrate dehydrogenase from porcine heart. Aluminum ions (Al3+) acted as a partial competitive inhibitor of the enzyme with respect to the substrate threo-Ds-isocitrate and inhibited the enzyme non-competitively with respect to NADP at pH 6.85. Fractional velocity plot analysis showed the Ki of the enzyme for aluminum ions to be 0.88 microM. When pH was elevated to 8.0, aluminum ions, which occur as a form of the Al(OH)4- anion, acted as partial uncompetitive and non-competitive inhibitors of the enzyme with respect to the substrates isocitrate and NADP, respectively. The Kí of the enzyme was determined to be 5.64 microM at pH 8.0 by fractional velocity plot analysis. The inhibition of NADP-isocitrate dehydrogenase by two forms of aluminum ions may explain aluminum toxicity in various tissues and organs.


Effects of histidine on tissue zinc distribution in rats.

Histidine has been reported to affect body zinc status by increasing urinary zinc excretion. The effects of experimental histidinemia on distribution of 65Zn in anesthetized rats were studied. Infusion of L-histidine at a rate sufficient to raise plasma concentrations to approximately 2 mM for 6 h starting 48 h after a single intraperitoneal 65Zn injection did not alter 65Zn activities in a variety of tissues when compared with anesthetized uninfused animals. However, plasma 65Zn and erythrocyte 65Zn were decreased, and liver 65Zn was increased. If 65Zn was injected intravenously during histidine infusion, net accumulation of zinc by some tissues was increased, but uptake by others was reduced relative to uninfused animals. In all cases, however, uptake expressed relative to plasma 65Zn levels was increased when allowance was made for the more rapid fall in plasma 65Zn during histidine infusion. Similar infusions of D-histidine produced quantitatively similar effects. Since enzymatic mechanisms and amino acid carriers would be expected to show stereoselectivity, such processes are unlikely to be involved in the zinc distribution changes described. The possibility of zinc transport by a hitherto unidentified carrier is discussed. These experiments confirm that histidinemia can affect zinc status, but any associated changes in urinary zinc excretion do not seem adequate to account for the tissue changes found.


Cadmium interferes with steroid biosynthesis in rat granulosa and luteal cells in vitro.

Recently, cadmium has been described to disturb ovarian function in rats. In this paper the direct influence of cadmium on steroid production of ovarian cells in vitro has been studied. Granulosa and luteal cells were obtained from proestrous and pregnant rats, and incubated with 0, 5, 10, 20 or 40 micrograms ml-1 CdCl2 in the presence or absence of 0.1-1000 ng ml-1 follicle stimulating hormone (FSH) or luteinizing hormone (LH) for 24 or 48 h. Production of progesterone (P) and 17 beta-estradiol (E2) by granulosa and that of P by luteal cells were measured by radioimmunoassay. In FSH-stimulated granulosa cell cultures, 5 and 40 micrograms ml-1 CdCl2 suppressed P accumulation to 65 and 10%, respectively; accumulation of E2 (at 5 micrograms ml-1 CdCl2) decreased to 44%. P production of LH-supported luteal cells dropped to 86 and 66%, respectively, when 5 and 40 micrograms ml-1 CdCl2 was added to the medium. No alteration in basal P accumulation occurred in granulosa and luteal cell cultures following incubations with 20 and 40 micrograms ml-1 CdCl2, whereas basal E2 production of granulosa cells was markedly diminished. It is concluded that CdCl2 suppressing steroid synthesis in vitro exerts a direct influence on granulosa and luteal cell function.


High performance liquid chromatographic determination of 3 alpha, 5 beta-tetrahydroaldosterone and cortisol in human urine with fluorescence detection.

A simple and sensitive method for the simultaneous determination of 3 alpha, 5 beta-tetrahydroaldosterone (THALD) and cortisol in human urine is described. The method uses high performance liquid chromatography with fluorescence detection. THALD and cortisol, released by enzyme hydrolysis, and fludrocortisone (internal standard) are isolated by a Sephadex G-25M column and a Bond-Elut C18 cartridge, and then oxidized by cupric acetate to form the corresponding glyoxal derivatives. The glyoxal derivatives are converted into the fluorescent quinoxalines by reaction with 1,2-diamino-4,5-methylenedioxybenzene. The quinoxalines are successfully separated on a reversed phase column (L-column ODS) with isocratic elution and monitored fluorimetrically. The detection limits for THALD and cortisol are 0.45 and 1.18 ng/mL urine (0.65 and 2.65 pmol/100 microL injection volume), respectively, at a signal-to-noise ratio of 3 in a 100 microL injection volume. This method permits the precise and sensitive determination of THALD and cortisol in human urine (2 mL).


Detection of gangliosides with the fluorochrome NBD dihexadecylamine and its application for preparative high performance thin layer chromatography.

A new method for the detection of gangliosides based on the lipophilic fluorescence agent 4-(N,N-dihexadecyl)amino-7-nitrobenz-2-oxa-1,3-diazole (NBD dihexadecylamine) and its application for preparative high performance thin layer chromatography is described. Brain gangliosides were chromatographed on silica gel coated thin layer plates and located with non-destructive fluorochrome under longwave ultraviolet light. The fluorescent zones were scraped off and the gangliosides were extracted with a mixture of chloroform/methanol/water (30/60/8; v/v/v). The gangliosides were separated from uncharged NBD dihexadecylamine by anion exchange chromatography and impurities were removed by Iatrobeads chromatography. The method described offers a simple and successful preparative thin layer chromatographic strategy to obtain pure gangliosides in microgram and milligram quantities.


A simple method for the quantification of famotidine in human plasma and urine by paired-ion high performance liquid chromatography.

A new method for the quantification of famotidine consists of a simple extraction procedure and paired-ion high performance liquid chromatography with ultraviolet detection. The method has good accuracy and precision and should be suitable for the routine measurement of plasma and urine samples for clinical studies.


High performance ion exclusion chromatographic characterization of the vaginal organic acids in women with bacterial vaginosis.

Vaginal organic acids have previously been detected by gas-liquid chromatography, but we have applied an ion exclusion high performance liquid chromatographic procedure to the analysis of vaginal discharge samples. This procedure has the advantage of not requiring derivitization of non-volatile acids and provides the convenience of a technique which does not require the use of flammable gasses, while allowing the identification of at least 18 different acids from the same chromatographic analysis. Vaginal discharge from women with symptoms of bacterial vaginosis was collected on weighed swabs and analysed for the presence of organic acids. The results were compared to the organic acid content of samples obtained from the same cohort of women after treatment with metronidazole. In addition, samples were obtained from asymptomatic women and these samples were analysed in the same manner. The number of organic acids present in samples from women with bacterial vaginosis was greater than the number found after treatment or among asymptomatic women. Succinic acid appeared to be inversely related to lactate concentration and succinate:lactate ratios were greater among women with bacterial vaginosis before treatment than after treatment. Liquid chromatography has proven useful as a means of evaluating the metabolic end-products of vaginal microorganisms in situ.


Quantitative determination of a new anticonvulsant (CGS 18416A) in human plasma using capillary gas chromatography/mass spectrometry.

An analytical method has been developed for the determination of a new antiepileptic drug, CGS 18416A, in human plasma. CGS 18416A is a new anticonvulsant representative of a novel class of water-soluble agents being developed for the treatment of epilepsy. Preclinical trials indicate sustained efficacy at relatively low oral doses, indicating a need for a sensitive assay. The method is based on capillary gas chromatography/mass spectrometry and utilizes stable isotope-labelled CGS 18416A as the internal standard. Samples (1 mL) are acidified, then washed with pentane/ethyl acetate, followed by liquid/liquid extraction at pH 11 with pentane/ethyl acetate. Extracts are then concentrated and analysed directly by gas chromatography/mass spectrometry. Separation is accomplished on a thick film methylsilicone capillary column. Mass spectrometry was carried out under positive ion ammonia Cl conditions with selected ion monitoring of the protonated molecular ions (m/z = 248 and 252) for CGS 18416A and the 13CD3-CGS 18416A, respectively. Specificity was demonstrated by the lack of interfering peaks at the retention time of CGS 18416A and the internal standard. Recovery and reproducibility assessments indicate good accuracy and precision over the validated concentration range of 0.2-51 ng/mL. The limit of quantification is 0.2 ng/mL and the method has sufficient sensitivity to support clinical trials. This is illustrated with an example of quantification in a normal volunteer following oral dosing.


A new method for the purification of cytochrome-P450 from human liver microsomes.

A procedure for the solubilization and purification of cytochrome-P450 (cyt-P450) from human liver microsomes is described. Successive treatment of microsomes with protease XXVII and 3-(3-cholamidopropyl)dimethylammoniopropanesulphonic acid gave a solubilized cyt-P450 in more than 80% yield and with a three-fold increase in specific activity. With this treatment it was possible to eliminate 80% of cytochrome-b5 and 75% of NADPH cyt-P450 reductase. The solubilized cyt-P450 was filtered on a Sephacryl-200 column and then subjected to high performance liquid chromatography with a Mono-P column (chromatofocussing). The recovery of separated cyt-P450 was about 50% with a specific activity of 11.5 nmol cyt-P450/mg protein. Also with this technique it was possible to determine the isoelectric points of cyt-P450. These results allowed us to confirm the usefulness of our method, for the study the cyt-P450 from surgical biopsies.


Detection of abnormal haemoglobin by capillary electrophoresis and structural identification.

Haemoglobin obtained from a male adult Ghanian with retinopathy, which was probably caused by haemoglobinopathy was analysed by capillary electrophoresis (CE) for clinical diagnosis. Two major peaks, which were in the ratio of nearly one, were detected. The elution times of these peaks (HbXI and HbXII) were faster than that of normal haemoglobin (HbA). The existence of two different abnormal types of haemoglobin was clear in the patient blood. The following sequence analysis revealed that the first peak (HbXI) was HbC and the second (HbXII) was HbS on the electropherogram, and that the patient was a heterozygote of HbS and HbC (HbSC disease). One of the diagnostic processes in a haemoglobin disease was shown by the combined use of CE, HPLC and a protein sequencer.


Determination of low molecular weight aliphatic primary amines in urine as their benzenesulphonyl derivatives by gas chromatography with flame photometric detection.

A selective and sensitive method for the determination of low molecular weight aliphatic primary amines in urine is described. These amines were converted into their benzenesulphonyl derivatives by a modified Hinsberg procedure, and measured by gas chromatography with flame photometric detection (FPD-GC) using a DB-1 capillary column. The derivatives were very stable and provided excellent FPD responses. By FPD-GC, linear calibration curves were obtained in the range 10-200 ng of methylamine, ethylamine, n-propylamine, isobutylamine and n-butylamine using tert-butylamine as an internal standard, and the detection limits of these amines were ca. 6-25 pg as the injection amount. Benzenesulphonamide derived from ammonia was converted into its N-dimethylaminomethylene derivative which has a longer retention time, and separated from benzenesulphonyl derivatives of low molecular weight primary amines on the chromatogram. The recoveries of aliphatic primary amines added to urine samples were 91-107% and the relative standard deviations were 0.2-4.5%. Analytical results of aliphatic primary amine contents in urine samples of normal subjects are presented.


The retention behaviour of conjugated bile acids in reversed phase high performance liquid chromatography.

The retention behaviour of conjugated bile acids has been studied in a reversed phase high performance liquid chromatographic (RP-HPLC) system by using the mixture of methanol and aqueous phosphate buffer as the mobile phase. The retentions of the conjugates in RP-HPLC have been found to be mainly controlled by the glycine and taurine groups. The selectivity between five different glycine and taurine conjugated bile acids is a constant in RP-HPLC. This selectivity has been used for peak identification in the practical separation of conjugated bile acids.


Venous leg ulcers: a prognostic index to predict time to healing.

To evaluate the prognostic factors in uncomplicated venous leg ulcer healing. Randomised parallel group controlled trial with subjects stratified by initial ulcer diameter and four months' maximum duration of follow up. Assessment at Northwick Park Hospital vascular unit with community based treatment. 200 patients with clinical and objective evidence of uncomplicated venous leg ulceration and an initial ulcer diameter > 2 cm. Time to complete healing of the ulcer. In the presence of graduated compression healing occurred more rapidly in patients with a smaller initial ulcer area (relative risk of healing 1.92 associated with halving of ulcer area (95% confidence interval 1.58 to 2.33)), shorter duration of ulceration (relative risk 1.35 associated with halving duration (1.17 to 1.56)), younger age (relative risk 1.34 associated with 10 year decrease (1.12 to 1.59)), and no deep vein involvement (relative risk 1.8 (1.19 to 2.78)). These prognostic factors used in a simple scoring system predict time to healing.


Oral cancer in Scotland: changing incidence and mortality.

To determine the incidence of oral cancer in Scotland between 1960 and 1989 and oral cancer mortality from 1911 to 1989. Data were obtained on oral cancer incidence from the information and statistics division of the Common Services Agency of the Scottish Health Service and mortality data from the office of the registrar general for Scotland. Mortality from intraoral cancers in Scotland substantially declined throughout this century until the mid-1970s. This trend, however, was then reversed, and fourfold increases in incidence were observed in younger age groups after 1960. Death rates in these younger age groups increased to levels previously recorded in the 1940s. These increases seemed to be cohort based and may therefore continue into the future. Reasons for increasing rates among younger age groups are speculative and rely on combining knowledge about risk factors and available ecological data. Though increases in incidence at younger ages do not result in a large change in the number of cases diagnosed, possible similar increases continuing into older ages, when oral cancer is more common, will correspond to a much larger increase in the actual number of cases. Given that such a large attributable risk is associated with tobacco and alcohol, however, these increases may be preventable.


Evidence for efficacy of drugs affecting bone metabolism in preventing hip fracture.

To examine the effects of taking drugs affecting bone metabolism on the risk of hip fracture in women aged over 50 years. Retrospective, population based, case-control study by questionnaire. 14 centres in six countries in southern Europe. 2086 women with hip fracture and 3532 control women matched for age. Number of drugs affecting bone metabolism taken and length taken for. Women taking drugs affecting bone metabolism had a significantly decreased risk of hip fracture. After adjustment for differences in other risk factors, the relative risk of hip fractures was 0.55 (95% confidence interval 0.31 to 0.85) in women taking oestrogens, 0.75 (0.60 to 0.94) in those taking calcium, and 0.69 (0.51 to 0.92) in those taking calcitonin. The fall in risk was not significant for anabolic steroids (0.6 (0.29 to 1.22)). Neither vitamin D nor fluorides were associated with a significant decrease in the risk of hip fracture. The effect on hip fracture risk increased significantly with increasing duration of exposure (risk ratio 0.8 (0.61 to 1.05) for less than median exposure v 0.66 (0.5 to 0.88) for greater than median exposure). Drugs were equally effective in older and younger women, with the exception of oestrogen. Oestrogen, calcium, and calcitonins significantly decrease the risk of hip fracture. Short term intervention late in the natural course of osteoporosis may have significant effects on the incidence of hip fracture.


Primary health care in London--changes since the Acheson report.

To examine changes in primary care in London in the 11 years since the Acheson report on primary health care in inner London. Analysis of key data from the family health services authority performance indicators and from the Department of Health; study of trends since the time of the Acheson report; examination of the provision of primary care in 1990-1 and its relation to health and social factors. Comparisons between the family health services authorities of inner London, outer London, and England as a whole, with a special study of Birmingham, Liverpool, and Manchester. The family health services authorities of England. There has been an improvement in the provision of primary care in inner London as judged by the criteria of the Acheson report, but these improvements have occurred only as part of an overall improvement in the provision of primary care in the country as a whole. None of the recommendations of the Acheson report specifically oriented to London have been implemented. There are some worrying trends in inner London, such as the increasing proportion of practices with more than 2500 patients. The problems faced by practitioners in inner London resemble those in other large inner city areas, but the primary care provision to deal with them is relatively poor.


Management of feverish children at home.

To compare the acceptability and effects on temperature of advice to unwrap children and give paracetamol or warm sponging treatments in the management of feverish illness at home. A randomised, open, parallel group study using factorial design comparison of unwrapping, warm sponging plus unwrapping, paracetamol plus unwrapping, and paracetamol and warm sponging plus unwrapping. Homes of willing families with a feverish child recruited after consulting one of 21 participating general practitioners in Southampton. 52 children aged from 3 months to 5 years with axillary temperatures before treatment of > or = 37.8 degrees C and < 40 degrees C. Response to advice assessed over four hours; temperature assessed by continuous data logging from an axillary thermistor; acceptability of treatment to child and parent scored on Likert scales immediately after treatment and on return to health. Response to treatment advice varied; unwrapping alone had little effect on temperature. Paracetamol increased the time below 37.2 degrees C in four hours by 109 (95% confidence interval 74 to 145) minutes compared with unwrapping; warm sponging caused the fastest reduction in temperature. Parents discriminated between treatments, preferring paracetamol. Advice to give paracetamol is more effective than sponging or unwrapping in controlling temperature in children at home and is more acceptable to parents. Warm sponging has an additive effect and reduces fever more quickly than paracetamol.


London after Tomlinson. Care in the capital: what needs to be done.

One of the aims of the Tomlinson report is to shift more care from the secondary to the primary sector in London. But the primary sector is already underresourced and overloaded. The capital has a heterogeneous population which often makes inappropriate demands on general practitioners. Many premises are inadequate and there are insufficient support staff. David Metcalfe emphasises that London is special and that the shift will not become a reality unless these problems are tackled. He suggests the establishment of different models of practice centres which could treat some of the patients who now go to accident and emergency departments. Some would be the night emergency service base, some would have primary care beds, and each would have a different mix of specialist support.


Faecal steroids and colorectal cancer: steroid profiles in subjects with adenomatous polyps of the large bowel.

In this necroscopy study the relation between carriage and size of colorectal polyps was correlated with luminal steroid concentrations in respect to malignant risk. Of the 92 subjects entered into the study, 68 had adenomatous polyps of the large bowel, of which 19 had adenomas > 0.9 cm in diameter (large adenomas), 26 in the range 0.5-0.9 cm in diameter (medium adenomas) and 23 of 0.4 cm or less in diameter (small adenomas). Sixty-three percent of subjects carrying large adenomas and 26% of persons carrying small adenomas had an abnormal ratio (> 1.0) of lithocholic acid to deoxycholic acid in intestinal contents as compared to 17% of the adenoma-free comparison group (n = 24). These findings support the suggestion that the ratio of lithocholic acid to deoxycholic acid as a faecal marker may be a useful adjunct to screening procedures for colorectal cancer.


Faecal steroids and colorectal cancer: the effect of lactulose on faecal bacterial metabolism in a continuous culture model of the large intestine.

The effect of lactulose on intestinal bacterial metabolism in two identical single-stage chemostats has been studied. The study was designed as a single stage crossover. Both cultures were inoculated and treated in an identical manner except that whilst one fermenter was subject to pH control the other was not and vice versa. Complex bacterial populations were maintained for 23 days and many of the metabolic functions carried out by the micro-organisms in vivo were demonstrated in vitro. The predominant organisms belonged to the genera Bacteroides, Bifidobacterium and Clostridium with abundant levels of anaerobic Gram-positive rods. The redox potential in each fermenter showed considerable variation with maximum values of below -300 eV being attained. An indication of the stability of the bacterial communities was the production of short-chain volatile fatty acids; during steady-state conditions the mean ratio of the major acids acetic, propionic and butyric being 3.90:0.69:1.00 and 3.65:0.76:1.00 in each fermenter, respectively. During steady-state conditions, 7 alpha-dehydroxylation of the primary bile acid chenodeoxycholic acid was maintained at a constant rate with lithocholic acid representing over 85% of total steroid. Addition of the soluble fibre lactulose to the cultures had a profound effect on many of the parameters tested. Without pH control the culture pH dropped to below 5.0 and this coincided with a substantial decrease in total anaerobes, especially Bacteroides spp., an increase in Lactobacillus spp. (concomitant with an increase in lactic acid), a decrease in the concentration of short-chain volatile fatty acids and a substantial decrease in 7 alpha-dehydroxylation of chenodeoxycholic acid. These results show that it is possible to maintain viable intestinal bacteria in vitro using a continuous culture chemostat and that the cultures respond to changing conditions as evinced by the inhibition of 7 alpha-dehydroxylation of chenodeoxycholic acid on addition of lactulose. This indicates that the model may be of further use in studying the modulation of secondary bile acid formation, a possible risk factor in colorectal carcinogenesis.


Biomarker assessments in asbestos-exposed workers as indicators for selective prevention of mesothelioma or bronchogenic carcinoma: rationale and practical implementations.

In the first part of this study we have shown how the serum levels of four selected tumour markers, namely tissue polypeptide antigen (TPA), carcino-embryonic antigen (CEA), hyaluronic acid (HA) and ferritin, display patterns characteristic of mesothelioma (M) or bronchogenic carcinoma (BC) in asbestos-exposed workers, and we hypothesize that the differences in marker patterns correspond to differences in carcinogenesis mechanisms. In a preliminary study, we found these specific marker patterns in 5/19 exposed workers of whom only one demonstrated any radiological signs of disease. Thus these specific marker patterns may be early events, occurring long (possibly years) before the classical radiological signs of exposure to asbestos. Accordingly they afford an optimal opportunity for prevention which should be adapted to the carcinogenesis mechanism as it is revealed by the marker pattern; it is aimed at antagonizing free radical carcinogenesis in all persons with TPA levels in excess of 100 U/l or Ferritin in excess of 400 ng/ml, and at inhibiting chemical carcinogenesis in those having elevated CEA levels (over 3 ng/ml). The mechanisms involved in these inhibitory processes are described and discussed, as well as the practical implementations that proceed from them. A prevention trial is now being started among 300 active and retired workers of an asbestos-cement works in northern France; the design of the study is presented. This prevention programme should be maintained over many years and holds a strong potential for reducing the untoward effects of exposure to asbestos.


Colorectal cancer and ischaemic heart disease: an uncommon inheritance!

This study reports the relative risk of death from cancer and from coronary artery disease in 1,811 first-degree relatives of 205 young colorectal cancer probands. The elevation in the risks of death from cancer (eg colon 3.6; rectum 2.0; uterus 1.8; cervix 2.3) is consistent with, though of lower magnitude than previous studies. An unexpected find was a highly significant deficit in coronary deaths. Recently discovered molecular associations between colon cancer and cholesterol metabolism suggest that further family studies of bowel cancer and heart disease in a variety of populations may be worthwhile.


Vitamin supplement use in a hospital-based population.

Vitamin supplement histories were taken from a sample of approximately 3,600 individuals (2,461 men and 1,181 women) from a hospital-based population. Multivitamin preparations were the most commonly used vitamin supplements (32% of men, 37% of women) followed by vitamins C (men 11%; women 16%) and E (men 17%; women 23%). The most common combination of supplements among the multi-users was vitamins C and E. Vitamin supplement use was positively associated with education in both sexes. Among women, there was a negative association between vitamin supplement consumption and Quetelet Index. No association was found between vitamin use and smoking status or alcohol use or amount of alcohol consumed.


Epidemiological aspects of cancer of the penis in Finland.

The purpose of this study was to characterize the epidemiological features of cancer of the penis in Finland. After histological re-examination and exclusions, the final series comprised 269 patients with cancer of the penis diagnosed and reported to the Finnish Cancer Registry between 1955 and 1977. The age-adjusted incidence rate of cancer of the penis decreased slightly from the 1960s onwards and was on the order of 0.5 per 100,000 person-years in the 1970s. Cancer of the penis was a disease of elderly males, which showed no systematic geographical variation in Finland in terms of risk. The most frequent predisposing factors were phimosis, in 44% of the cases, and condylomatous lesions, in 20% of the cases. The general survival (all patients, all stages) was favourable; the 5-, 10-, and 20-year relative survival rates were 71, 69 and 78%, respectively.


Incidence of cancer of oesophagus, cardia and stomach in Denmark.

Subtypes of oesophageal and gastric cancer in Denmark are compared with respect to their occurrence in men and in women, and in the Danish capital, Copenhagen, versus the rest of Denmark. Three categories of tumours can be distinguished epidemiologically: oesophageal squamous carcinoma, oesophageal and cardial adenocarcinoma, and distal gastric cancer. Comparison of information reported to the Danish Cancer Registry and cause of death as indicated on death certificates suggests that considerable misclassification between subtypes of tumours occurs. The Danish experience supports the findings from other populations of increasing incidence of oesophageal and cardial adenocarcinoma, but because of possible changes in diagnoses over time and of misclassification of subtypes, the data must be interpreted with caution.


Female tumours in Italy: an example of quantitative assessment of disease burden and preventive strategies.

Female tumours (ie cancers of the female genital tract and breast) represent alone about 40% of new cases and 30% of cancer deaths in Italy (approximately 44,500 new cancer diagnoses and 17,800 deaths in 1991). An attempt to identify major issues in the primary and secondary prevention of these malignancies has disclosed several important points. Firstly, concerning epidemiology, excesses of mortality from cancers of the breast and ovaries exist in the northern part of Italy as compared to the centre and south, whereas mortality rates from uterine cancer are almost equally distributed. However, a tendency towards increasing similarity of various Italian regions emerges. Risk factors most amenable to intervention are overweight, contraceptive methods and dietary habits. Secondly, concerning screening, the number of mammograms (about 552) and the number of Pap smears taken every year (about 4 million) are approximately sufficient to provide a complete coverage of the target subset of the female population as concerns breast and cervical cancer screening. Still, on account of no one carrying out primary call procedures and checks, the actual situation is unsatisfactory. The present article, in addition to quantifying the present status of female cancer prevention in Italy, tries to define a framework for similar exercises in other European countries.


Plasma vitamin concentrations in patients with intestinal metaplasia and in controls. UK Subgroup of the ECP-EURONUT-IM Study Group.

A case-control study has been carried out on 129 patients with newly diagnosed intestinal metaplasia (IM) recruited in four UK centres (London, Slough, Stoke and Glasgow). Serum assays of a range of vitamins demonstrated that 38% of IM patients had serum ascorbic acid levels below the detection levels compared with only 16% of controls free of chronic gastric lesions; this difference was also seen in the patients from the individual centres. There were no major differences between cases and controls in any other serum vitamin levels measured (carotene, tocopherol or the ratio of tocopherol to cholesterol). Comparison of the healthy asymptomatic controls suggested that the intakes of ascorbic acid and of beta-carotene were much higher in southern England than in Stoke or Glasgow. The tocopherol status in the four centres tended to be more similar.


Aetiology of oesophageal cancer: some operative factors.

Of the thousands of chemicals tested, the only compounds found to be potent carcinogens for the oesophagus are the N-nitrosamines. Many of these compounds are readily formed from common precursors in the environment (eg example in food during its storage or preparation) and also in vivo in the human stomach. Exposure is therefore likely to be ubiquitous. Although man may be exposed to other oesophageal carcinogens these have yet to be chemically identified, and at present nitrosamines are the sole contenders for the role of initiators of oesophageal cancer in man. Evidence suggests strongly that oesophageal cancer is initiated world-wide by nitrosamines, and promoted by secondary factors, the nature of which varies with the population concerned, notably alcohol in Europe and the USA, dietary deficiencies in China and Iran, mycotoxins in South Africa. When several risk factors coincide in one locality, the result can be a very high incidence of oesophageal cancer, with no one major cause.


Lactobacilli, anticarcinogenic activities and human intestinal microflora.

Lactobacilli belong to the normal oropharyngeal and intestinal microflora in humans. These microorganisms contribute to the stabilization of the microflora and maintain the colonization resistance against pathogens. Lactobacilli have been used as dietary supplements in order to prevent gastrointestinal disturbances. Claims have been made that certain strains of lactobacilli possibly exert anticarcinogenic activities. The activity of bacterial enzymes, implicated in colon carcinogenesis may be elevated by a high meat, Western-type diet. Supplements of Lactobacillus acidophilus decreased these levels in both rats and humans. Colon cancer patients given L. acidophilus fermented milk showed a significant increase both in numbers of intestinal lactobacilli and dietary calcium intake, while decreasing trends in levels of both soluble faecal bile acids and faecal bacterial enzymes, two risk makers for colon cancer, were observed. In vitro studies have revealed that lactobacilli and other lactic acid bacteria have the ability to absorb cooked food mutagens. Recent studies in humans have shown that intake of L. acidophilus significantly reduced the mutagen excretion after consumption of fried meat. Several mechanisms by which lactobacilli might exert anticarcinogenic effects are discussed. Thus, certain strains of lactobacilli might lower the colon cancer risk in humans.


A food fair to promote dietary change in the Stockholm cancer prevention programme.

Involving organizations is crucial when developing community intervention strategies aimed at dietary change. This case study describes the use of a food fair as a change agent. The aim of the food fair was to obtain commitment and build up a network in the community comprised of professionals in the fields of food production, food distribution and information. The fair was organized in 1989 and intended to disseminate knowledge and information about healthy foods and encourage food industries, test kitchens and also publishing companies to develop and present products in accordance with the dietary objectives of the Stockholm Cancer Prevention Programme (ie a simultaneous reduction of fat and an increase in fibre intake). The food fair consisted of exhibitions and conferences/seminars. Policy makers, journalists, people in the educational and health professions and those working with food were invited to participate. Nearly 60 companies and organizations made presentations at the exhibition and 1,000 professionals attended the conferences and seminars. In all, 6,400 people visited the fair. Seventy-eight trade journals and newspapers and three radio stations reported on the fair. The food fair was so well received that a second fair was arranged in 1991.


A tobacco quit and win model in the Stockholm cancer prevention programme.

The main objective of the Stockholm Cancer Prevention Programme (SCPP) is to reduce cancer incidence and mortality among the 1.6 million inhabitants in Stockholm county by reducing risk factors particularly related to lifestyle. The objective of the SCPP's tobacco action programme is to reduce the number of adult daily users of tobacco (including oral snuff) to 20% by the year 2000. In 1988, a nationwide Quit and Win contest was conceived as part of this long-term programme. The contest recruited nearly 13,000 participants or 0.6% of the daily tobacco users in Sweden over the age of 16 years. More than 60% of the participants were recruited from Stockholm county. This corresponds to 1.9% of the daily tobacco users in Stockholm county compared with 0.3% in the rest of Sweden. The pharmacies and the public health services organizations were the principal distributors of contest entry forms. Sixty-two percent of the men and 59% of the women were tobacco free one month after the contest, and after 6 months the corresponding figures were 30 and 25%, respectively.


Tobacco and cancer: how to react to the evidence.

Tobacco is a remarkable product when considering the number of diseases to which it can reliably be linked as a causal agent or at least as a factor which is partially responsible for the occurrence of disease. If we only look at cancer, tobacco is responsible for the vast majority of cancers of the lung, trachea and bronchus; a proportion of cancers of the bladder and of the renal pelvis; a considerable proportion of cancers of the oral cavity, lip, pharynx, larynx and oesophagus with, for these sites, a large potentiation of risk when associated with the drinking of alcohol; a proportion of cancers of the pancreas and possibly of renal adenocarcinoma. In addition, among women, tobacco use has been linked with an increased risk of cervical cancer. In order to evaluate the present burden of tobacco-related cancers in the world precisely, one needs to be able to consult precise statistics of cancer occurrence, tobacco use and the relationship between the two. The availability of such data varies greatly across countries. Although mortality, and to a lesser extent, morbidity cancer statistics are collected in many parts of the world, the coverage is far from exhaustive and of uniform quality. Commercial or governmental sources of data on tobacco also exist, but the most important missing link is knowledge of the population-specific risk attributable to tobacco use in various countries for all tobacco-related diseases. Despite imperfect knowledge, the link between tobacco use and cancer is clear enough to demand strong action.(ABSTRACT TRUNCATED AT 250 WORDS)


The reliability and validity of a Dutch questionnaire on fat consumption as a means to rank subjects according to individual fat intake.

In this paper a short, 25-item, telephone-administered questionnaire, designed to rank individuals according to their dietary intake of fat, is presented. Based on two studies of 52 and 639 men and women, respectively, its relative validity regarding a 7-day diet record and its reproducibility are discussed. A Pearson correlation of 0.59 was observed between fat intake estimated by 7-day diet records and fat score derived from the short questionnaire. Gross misclassification, defined as disagreement between the two fat consumption assessments beyond an adjacent tertile, was 15.4%. A Pearson correlation of 0.71 was calculated by a test-retest procedure. It is concluded that the Dutch short questionnaire on fat consumption is a rapid, simple and inexpensive method that can very well be used in nutrition education research to rank individuals according to their dietary fat intake.


The effect of colonic site of biopsy on cell proliferation profiles using BRDU incorporation as a measure of cell proliferation.

Sigmoidal and rectal biopsies were taken from 10 patients at colonoscopy. Colonic crypt cell proliferation was measured using a monoclonal antibody to bromodeoxyuridine by an immunohistochemical technique. Labelling index and crypt cell proliferation profiles were analysed and compared between the two sites. No statistically significant difference was found in proliferation parameters. Assessment of colonic proliferation is a useful indicator for colorectal cancer risk and for determining the effects of ongoing drug treatment. Rectal biopsies are easier to take than sigmoidal biopsies. The results of this study suggest that there is no difference in cell proliferation at the two sites. Rectal biopsies are more practical for clinical studies.


Preventive intervention in breast cancer, but when?

After an introductory note on primary preventive intervention of breast cancer during adulthood, the author defends and extends a hypothesis that relates most of the known risk factors for this disease to the development of preneoplastic lesions in the breast. If changes in lifestyle concerning nutritional and/or reproductive risk factors during puberty and adolescence would turn out to be unacceptable socially, an alternative approach might be found in chemoprevention based on cell differentiation.


Growth of colorectal polyps: design of a prospective, randomized, placebo-controlled intervention study in patients with colorectal polyps.

One hundred and sixteen patients were included, during 18 months, in a double-blind placebo-controlled intervention study, with calcium, vitamins A, C, E and selenium (in a cocktail) or placebo against growth of colonic polyps. Patients were randomized within three arms, according to diameter of the largest polyp, < 5 mm, 5-9 mm or > 9 mm. Polyps > 9 mm were resected, the others were left to be measured annually before resection after 3 years. The protocol (performed in all of the patients) included registration of demographic data, family and personal history, measurement of polyps, collection of blood specimens, stools and biopsy samples. Registration of nutritional status, diet history and 5-day prospective food consumption, was performed in 108 patients. The patient compliance was registered every third month by the hospital pharmacist, with concomitant delivery of new boxes of capsules. Additionally, stool collections were performed from all of the patients for the measurement of faecal calcium, bile salts and fat. Inclusion rate of 37, 41 and 38 patients in each of the three 6-month periods was uniform. The group with the largest polyps measuring 5-9 mm comprised 44% of the material. The sex ratio corresponded to that in overall referrals for colonoscopy. The age relationship of size and multiplicity of polyps and the distribution of polyps in the large bowel corresponded to previous experience in polyp-bearing individuals of the same age.(ABSTRACT TRUNCATED AT 250 WORDS)


Cell proliferation study on gastric carcinoma and non-involved gastric mucosa using a bromodeoxyuridine (BrdU) labelling technique.

Previous reports using [3H]thymidine have shown altered patterns of cell proliferation in gastric carcinogenesis. In this study we applied in vitro bromodeoxyuridine (BrdU) uptake and immunohistochemistry to tissue from carcinoma, its adjacent mucosa and distant mucosa (n = 16) cases and 10 normal control cases. The labelling index (LI) was calculated for the upper, neck/isthmus and lower gland compartments and in 10 random fields/carcinoma. The LI was greater in the carcinoma than in either the adjacent or far mucosa or the controls (p < 0.001). No difference was observed between the non-carcinoma areas. However, the LI varied significantly between the gland compartments within each area (p < 0.001). In the normal control cases, the location of the proliferating cells was limited to the neck/isthmus region and perhaps the most significant observation was the change of location and expansion of the proliferative zone towards the surface in the normal mucosa adjacent to carcinoma compared with distant mucosa and controls (p < 0.001). These altered patterns of cell proliferation may be an important aspect of the precancerous potential of field changes in gastric carcinoma patients and could help patient follow-up and serve as a marker in monitoring intervention studies.


The relationship between gastric mucosal changes and nitrate intake via drinking water in a high-risk population for gastric cancer in Moping county, China.

There are conflicting reports on whether nitrate intake is related to gastric carcinogenesis. In this laboratory/field study from a high-risk area for gastric cancer, we analysed 178 samples of drinking water for nitrate and nitrite, and examined the relationship between gastric mucosal lesions (including gastric cancer) and quality of different types of drinking water and nitrate intake via water. The results showed that the nitrate content in the local drinking water was generally very high, with a mean of 109.6 mg/l (range 4.4-497.2 mg/l). There were significant differences in the nitrate content in drinking water from different wells in qualitatively different types of water. The histological changes were closely related to the quality of drinking water and its nitrate content. The results suggest that nitrate in drinking water probably plays an important role in gastric carcinogenesis and that in future aetiological studies of gastric cancer should include more information on well depth, the presence of public or private wells and nitrate content of water.


Acute megakaryoblastic leukemia in children: treatment with bone marrow transplantation.

Seven children underwent BMT for acute megakaryoblastic leukemia (AMKL). They were assessed for clinical, hematologic, and cytogenetic findings as well as response to treatment. The diagnosis of AMKL was established by cytochemistry, immunophenotyping and/or platelet-peroxidase reactivity. Patients had received various prior chemotherapies. One was in first remission, another in second remission and five were in relapse at the time of admission for transplant. Marrow donors included an HLA identical sibling (one), phenotypically HLA identical unrelated (two) and partially HLA identical family members (four). Five patients achieved engraftment, one rejected the graft and died on day 20 after a second unrelated transplant and one died from infection on day 5. Two patients relapsed within the first month after transplant and died of recurrent leukemia. Another died of a second malignancy on day 2232. Two patients survive disease-free more than 3.8 and 4.3 years after transplant.


High dose chemotherapy with ABMT in soft tissue sarcomas: a report of 22 cases.

Twenty-two patients with locally advanced or metastatic soft tissue sarcomas received high dose chemotherapy with autologous bone marrow graft. Eleven patients receiving melphalan also received fractionated total body irradiation. Six patients (four in CR and two in PR) were intensified after first line therapy. Thirteen patients were grafted after chemosensitive relapse: seven in second CR, one in third CR, one in first PR, three in second PR and one in fourth PR. Three patients with primary refractory disease were intensified. The overall response rate in 66% in nine evaluable patients. The overall median survival and disease-free survival were 19 and 15 months, respectively. The actuarial survival rates at 2 and 5 years were 40% and 32% respectively. There was one treatment-related death due to infection. We conclude that high dose chemotherapy is feasible and provides reasonable response rates in patients with advanced soft tissue sarcomas.


Peripheral blood stem cell collection and autografting in high risk lymphomas.

Peripheral blood stem cells (PBSC) were collected from 29 patients with high risk Hodgkin's disease (n = 3) or non-Hodgkin's lymphoma (n = 26) in partial remission or first sensitive relapse. Patients had either bone marrow involvement or hypoplastic bone marrow. The conditioning regimen prior PBSC collection included amsacrine and cytosine arabinoside (Ara-C) or Ara-C alone. PBSC collection was performed after leukocyte counts reached 1 x 10(9)/1. A good yield was obtained in 23 patients, whereas sufficient numbers of CFU-GM were not obtained in six cases. Twenty-one patients have been transplanted. All patients except one achieved bone marrow engraftment. Eight patients are in complete remission (mean duration 15 months). The estimated 2 years survival rate is 46.4% (CI 25-68%). This procedure would seems a good alternative in poor prognosis lymphomas.


Recombinant human erythropoietin (rh-Epo) administration to normal marrow donors.

Ten normal marrow donors, two children and eight adults, received 9-22 daily subcutaneous doses (100 units/kg) of recombinant human erythropoietin (rh-Epo) and oral iron prior to marrow harvesting. The two children did not have autologous blood stored prior to marrow harvesting while all eight adults did. Except for a mild skin rash at the sites of injection in three cases and transient headaches in a further three, no side effects were observed. The effects of rh-Epo on preoperative and postoperative hematocrits were evaluated and compared with those of control donors matched for age, sex, weight and blood loss. Initial hematocrits were assigned a value of 100%. The mean percentage increase between the initial hematocrit and the preoperative hematocrit in the rh-Epo group was 16% compared with a decrement of 4% in the control group (p = 0.0001). The mean % decrement between the initial hematocrit and the postoperative hematocrit was 4% for the rh-Epo group and 26% for the control group (p = 0.0003). It was concluded from this study that rh-Epo could be given safely to normal marrow donors with a significant increase in hematocrit occurring in the 2-3 weeks prior to marrow harvesting. This approach should be explored further, especially in children, for whom storage of autologous blood is not routinely performed.


Early deaths in children after BMT. Bone Marrow Transplantation Group of the Italian Association for Pediatric Hematology and Oncology (AIEOP) and Gruppo Italiano Trapianto di Midollo Osseo (GITMO).

In order to determine the incidence and causes of death during the first 100 days after BMT (early deaths) in a pediatric population we have examined data reported in the AIEOP BMT Registry. Up to July 1990, data on 486 children who underwent allogeneic (180) or autologous (306) BMT were evaluable. The children had acute lymphoblastic leukemia (148 cases), acute non-lymphoblastic leukemia (127 cases), neuroblastoma (82 cases), chronic myelogenous leukemia (15 cases), aplastic anemia (nine cases), solid tumors, lymphoma, immunodeficiency or storage diseases. The overall survival is 55% for allogeneic HLA matched and 38% for autologous transplants at 5 years, 24% for HLA mismatched graft at 2 years. Out of the 486 children, 70 (14%) died during the first 100 days after BMT: 33/306 (11%) after autologous BMT, 24/150 (16%) after allogeneic matched BMT and 13/30 (43%) after mismatched BMT. Causes of early death were as follows: disease progression: 12 children (10/306 after autologous and 2/180 after allogeneic BMT); infection: 12 children (five after autologous and seven after allogeneic BMT); interstitial pneumonitis: 21 children (seven after autologous and 14 after allogeneic BMT); cardiac failure: five children (four after autologous BMT); veno-occlusive disease: eight children (three after autologous, five after allogeneic BMT); acute renal failure: three children (one after autologous and two after allogeneic BMT); multiple organ failure: two cases (one after autologous BMT); cerebral hemorrhage: three children (one after autologous BMT); hypertension: one child; acute GVHD: three children (12% of early deaths after allogeneic BMT).


Effect of radiation dose on the development of mixed haemopoietic chimerism following T cell-depleted allogeneic bone marrow transplantation.

The presence of mixed haemopoietic chimerism (MXC) was evaluated by cytogenetic and molecular analysis in 48 patients undergoing T cell-depleted BMT. The dose of total body irradiation (TBI) prescribed to all patients (14.4 Gy) was calculated to compensate for the absence of T cells in the graft. The actual midline dose of TBI received, however, differed significantly depending on the method of TBI administration. Thus, 35 adult patients received an average midline dose of 14.3 Gy, while 13 children received a lower dose of 13 Gy. The incidence of MXC in the adult group, who had received very close to 14.4 Gy to the midline, was 34% (12/35), which is lower than in most reported T cell-depleted series. During follow-up, chimerism remained relatively stable with time but varied between haemopoietic lineages. There was no relationship with relapse. MXC in the 13 children who had received a lower midline TBI dose was significantly higher at 69% (9/13) (p < 0.05) and increased to 90% (9/10) if patients who received additional chemotherapy in their conditioning were excluded (p = 0.001). This suggests that, in terms of marrow ablation, relatively small changes in the dose of TBI may be biologically significant, at least at this dose range. Again, in the lower TBI group MXC was not predictive of relapse.


Effective prevention of acute GVHD following allogeneic BMT with low leukaemic relapse using methotrexate and therapeutically monitored levels of cyclosporin A.

Although the combination of cyclosporin A (CYA) and methotrexate has been reported to reduce the incidence of acute GVHD in patients undergoing allogeneic BMT for leukaemia, it has been associated with a higher risk of leukaemic relapse. Since 1987 we have used the combination of CYA and methotrexate for GVHD prophylaxis in 24 patients undergoing allogeneic BMT for leukaemia or myelodysplasia. Over the first 50 days post-transplantation, CYA dosage was adjusted to keep within a therapeutic range of 95-205 ng/ml. This resulted in a 60% reduction in CYA dosage by day 50 post-transplant compared to the original Seattle protocol. Despite the low dosage of CYA administered, the incidence of acute GVHD was only 25% with no patient having greater than grade I GVHD. There have been no leukaemic relapses in low risk patients. The results indicate that decreasing CYA dosage does not increase the incidence of GVHD but may reduce the risk of leukaemic relapse following allogeneic BMT.