The effect of deliberate omission of Trinordiol or Microgynon on the hypothalamo-pituitary-ovarian axis

The effect of deliberate omission of a phased formulation pill, Trinordiol (ethinyl estradiol 30 micrograms + levonorgestrel 50 micrograms: 6 tablets; ethinyl estradiol 40 micrograms + levonorgestrel 75 micrograms: 5 tablets; ethinyl estradiol 30 micrograms + levonorgestrel 125 micrograms: 10 tablets) or a low-dose, combined, oral contraceptive pill, Microgynon (ethinyl estradiol 30 micrograms + levonorgestrel 150 micrograms: 21 tablets) on the hypothalamo-pituitary-ovarian axis were studied. Thirty-six women were recruited to the study and divided equally between the two types of pill. Medication was begun on the 8th pill-free day of the cycle and continued for 7 days (Group 1), 14 days (Group 2) or 21 days (Group 3). Levels of FSH, LH, estradiol (E2) and progesterone (P) were measured in plasma on alternate days during the final week of pill therapy, and daily for the 7 days after stopping the pill. For the first 2 weeks of pill therapy, follicular activity, as judged by plasma levels of E2, was greater in women taking Trinordiol than in those taking Microgynon, but was similar in both groups by the third week of pill treatment. Five women taking Trinordiol (2 in Group 1 and 3 in Group 2) had plasma levels of E2 in excess of 500 pmol/l whilst taking the pills, and only 1 patient achieved this degree of follicular activity after stopping the tablets. One woman who had taken 7 days of Trinordiol (Group 1) showed a rise of plasma levels of P to 6.8 nmol/l, but luteinization did not occur in any of the remaining 35 women who took Trinordiol or Microgynon. These findings suggest that follicular activity is less completely suppressed by Trinordiol than Microgynon, at least in the first 2 weeks of pill therapy, but that normal ovulation is still a rare event in the week after cessation of either of these pills, even if only 7 days of medication have been taken.     

Pituitary, ovarian and endometrial effects of 300 μg norethisterone and 30 μg levonorgestrel administered on cycle days 7 to 10

The ovarian, endometrial and pituitary effects of 300 μg norethisterone (NET) and 30 fig levonorgestrel (L-NOG) administered orally on cycle days 7–10 were investigated in two groups of 10 women each, by daily analysis of plasma estradiol (E₂), progesterone (PROG), immunoreactive luteinizing hormone (LH) and follicle stimulating hormone (FSH) in a pretreatment control cycle and during NET or L-NOG administration. Endometrial biopsies were obtained for morphometric analysis on cycle day 11 in the control and treatment cycles. Treatment with 300 μg NET resulted in an increase in the area under the E₂ peak (p<0.05), reduction in the number of subjects with normal progesterone profile (p<0.05) and a decrease in the area under the progesterone curve (p<0.05). The treatment suppressed the LH peak in 4 subjects and progesterone in 4 subjects. The follicular phase was prolonged in one subject, Norethisterone induced marked subnuclear vacuolation in the endometrium, while the glandular mitoses were decreased during NET treatment. Treatment with 30 μg L-NOG resulted in a decrease in subjects with normal progesterone profiles (p<0.05) and in the area under the progesterone curve (p<0.05). The treatment suppressed the LH peak in 3 subjects and progesterone in 4 women. The follicular phase was prolonged in one subject. L-NOG did not significantly increase the diameter of glands or induce subnuclear vacuolation in the endometrial glands.     

The “U” technique: A new method for Norplant implants removal

The standard technique for Norplant implants removal was compared with a new technique called the “U” technique, which employs the use of a modified no-scalpel vas deferens holding forceps to grasp and remove the capsules. Seventy-six women requesting Norplant implants removal were randomly assigned to Group 1 (standard removal technique) or Group 2 (“U” removal technique). Variables measured included: (a) time required for complete removal, (b) number of capsules not broken or damaged during removal, and (c) number of incisions required for removal of all six capsules. In clients with visible or palpable capsules, the standard removal technique required significantly more time to remove all six capsules, on average, than the “U” technique (19 versus 7 minutes, p < 0.001); also more capsules were damaged during removal (5.6 undamaged versus 6.0,p < 0.01). In addition, with the standard technique, five clients required two incisions for removal of all six capsules while none of the clients required more than one incision with the “U” technique. Our conclusion is that the “U” technique is a quicker and easier method of removing Norplant capsules than the standard technique.     

Pituitary-ovarian activity when switching from various monophasic pills to a triphasic pill

This study was prompted by the various recommendations given by different oral contraceptive manufacturers to women who wish to switch from a monophasic to a triphasic formulation. Ten women who switched from a variety of monophasic pills to a levonorgestrel triphasic pill formulation after a 7-day pill-free interval were studied. Follicular maturation was monitored by ultrasound scan, and the levels of serum follicle stimulating hormone (FSH), luteinizing hormone (LH), progesterone (P) and estradiol (E₂) measured. Pituitary-ovarian activity was suppressed in six of the ten women studied, while in the remaining four women there was some pituitary-ovarian activity during the first 10 days on the triphasic pill. These findings suggest a shorter pill-free interval, as advised in USA data sheets, may be less likely to result in pill failures when women switch from a monophasic to a triphasic preparation.     

“pop-out” method of levonorgestrel implant removal

The “pop-out” technique is a method of levonorgestrel implant removal that uses digital pressure to direct implants through a small skin incision. This technique was developed, theoretically, to cause less bruising and patient discomfort by avoiding the use of instruments. The pop-out technique is the primary method used for levonorgestrel implant removal in the Magee-Womens Hospital resident clinic. We performed a retrospective analysis of levonorgestrel implant removals performed between July 1, 1995, and December 31, 1998. Of the 168 removals included in this analysis, 38 were performed by one of two attending physicians, and 130 were performed by the residents with attending supervision. The average time for removal was 12 ± 5 min (range 2.25–27 min) when the “pop-out” method could be used to remove all six implants, and 14 ± 7 min (range 2.25–59 min) for all removals. The removal time for residents was inversely proportional to the anticipated level of difficulty of the removal and to the number of previous removals performed. The removal time was significantly faster when residents were supervised by one of the attending physicians as compared with the other attending physician. Only 0.7% (7/1,008) of levonorgestrel implants were fragmented during removal. This review shows that the “pop-out” method is a reasonable alternative to other proposed methods of primary implant removal. The difference in the level of expertise of the attending physician may significantly influence removal time when training clinicians in levonorgestrel implant removal.     

Pharmacodynamic effects of ethinyl estradiol in women using vaginal devices releasing small doses of levonorgestrel at a constant rate

Eight normally menstruating women were provided with vaginal devices releasing levonorgestrel (NOG)4) at a constant rate of 20 micrograms/24 h. On day 71 or 72 following the insertion of the device, oral doses of 50 micrograms of ethinyl estradiol (EE) were administered daily for one week. Peripheral blood samples were drawn three times weekly during a pretreatment (control) cycle and from day 29 of the treatment period. The levels of progesterone (P), estradiol (E2) and NOG were measured by radioimmunoassay, sex hormone binding globulin (SHBG) by a steady state polyacrylamide gel electrophoresis and the percentage of binding of NOG, testosterone (T) and E2 by equilibrium dialysis of diluted plasma. An endometrial smear and a biopsy were taken from each subject on 3 occasions, viz. during the control cycle (cycle day 20-22), during the period with the NOG-releasing device in situ (44-50 days after the insertion of the device), and on the 7th day of concomitant EE administration.     

Effects of different doses of norethisterone on ovarian function, serum sex hormone binding globulin and high density lipoprotein-cholesterol

The ovarian effects of different doses of norethisterone (NET) were compared in 45 normally menstruating women in order to find the lowest effective dose of the Chinese NET “visiting pill”. Subjects were randomly divided into 3 groups. Each subject in each group was taking 0.5, 1.5 or 3.0 mg per day from days 5 to day 18 of the cycle. Blood samples were taken on days 5, 8, 11, 14, 17, 20, 23, 26 and 29 of the cycle. Serum oestradiol (E₂), progesterone (P), sex hormone binding globulin (SHBG), high density-lipoprotein cholesterol (HDL-C), and NET concentrations were measured. Ovulation, delayed ovulation, ovulation inhibition and follicular activity were classified by the analysis of the peripheral serum levels of sex hormones. Ovulation occurred in 7 women in the 0.5 mg group, in 2 women in the 1.5 mg group and in none of the 3.0 mg group. Mean serum SHBG levels were reduced progressively by 6.6% (Group 0.5), 15.5% (Group 1.5) and 23.4% (Group 3.0). There were no significant changes in HDL-C levels in any group. There was a significant correlation of mean serum NET concentrations with dose. The lack of complete inhibition of ovulation in most women in the 1.5 mg and 0.5 mg groups might suggest that the dose of NET required when used as a visiting pill could not be reduced below 3.0 mg.     

A clinical pharmacological study of a new type of vaginal delivery system for levonorgestrel

The pharmacokinetic and pharmacodynamic effects of a new type of levonorgestrel-releasing vaginal device (with an in vitro release rate of 25 μg/24 h) were studied in a group of 18 normally menstruating women during a period of 90 days of continuous use. Peripheral blood samples were drawn three times weekly (Mondays, Wednesdays, Fridays) during a pretreatment (control) cycle and during the 90 days (3 segments) of exposure to levonorgestrel and the levels of levonorgestrel, progesterone and estradiol were analyzed. Blood samples were also drawn at frequent intervals during the first day and daily during the first week with the devices in situ. In addition, endometrial biopsy specimens were obtained during days 20–22 of the control cycle and then 6 and 10 weeks following the insertion of the devices for morphometric analysis and for the assay of progesterone and estradiol levels.

Following insertion of the devices, plasma levels rapidly rose to 1 nmol/l in 12 hours; a plateau of approximately 1.6 nmol/l was reached in 3–4 days, after which the plasma levels declined in a linear fashion with a daily average rate of 7.4 pmol/l to 60% of the initial level in 90 days' time. Of the 54 treatment segments of 30 days, 685 were anovulatory and 24% showed normal, ouvlatory-like estradiol and progesterone levels.

A complete set of three biosies were obtained from 15 of the 18 subjects. Of the biopsies obtained during exposure to levonorgestrel only one exhibited signs of atrophy, 80% showed suppressed or arrested proliferation, and 10% had a normal cyclic appearance. Morphometric analysis revealed a significant (p < 0.01) decrease in the number and diameter of endometrial glands and an apparent increase in the number of biopsies exhibiting glandular and/or stromal mitoses during the treatment period.

Although — compared to control biopsies — apparently decreased progesterone and increased estradiol levels were found in the biopsy specimens taken during treatment, there was no correlation between endometrial and peripheral steroid levels; in 10 “treatment” biopsies high endometrial progesterone levels were found in the presence of very low (< 1.0 nmol/l) peripheral plasma levels.

An assessment of the bleeding profiles during exposure to leveonorgestrel indicated that 35 (geometric mean) and 39 (arithmetic mean) per cent, respectively, of the treatment days were days with bleeding or spotting. No difference was found in this respect between anovulatory and ouvlatory-like segments.

It is concluded that the devices studied seem to cause too many bleeding irregularities. It is suggested that the development of improved “progestogen-only” contraceptives hinges on a better understanding of the mechanism of antifertility effect of low-dose progestogens and of the intermenstrual bleeding caused by them.     

Pituitary,ovarian and endometrial effects of progesterone released prematurely during the proliferative phase

The pituitary, ovarian and endometrial effects of premature exposure to progesterone during the proliferative phase were investigated in 18 normally menstruating women. Daily blood samples were drawn during a control cycle and an endometrial biopsy was obtained on cycle day 6 (in 9 subjects) and on cycle day 11 (in 9 subjects), respectively. Daily blood samples were drawn again throughout a treatment cycle, in which a vaginal delivery system releasing progesterone at a constant rate of 1.4 mg/24 h was inserted and left $\text{in situ}$ for 96 hours during cycle days 2–6 (9 subjects) and cycle days 7–11 (9 subjects), respectively. On the 6th and 11th cycle day, respectively, the devices were removed and another biopsy specimen was obtained. In all blood samples the levels of immunoreactive lutropin (LH), estradiol (E₂), 17-hydroxyprogesterone (17-HO-P), progesterone (P) and 20α-dihydroprogesterone (20α-HO-P) were estimated by radioimmunoassay.Insertion of the devices resulted in a rapid, approximately six-fold increase (from 0.5 to 3.0 nmol/l) in P levels and an approximately threefold (from 0.5–0.6 to 1.5–2.0 nmol/l) increase in 20α-HO-P levels. A high degree of correlation was found between P and 20α-HO-P levels. No major changes were observed in the profiles and levels of the other hormonal indices studied.However, premature exposure to small amounts of P during cycle days 2–6 resulted in a significant decrease in the ratio of the E₂ to LH peaks and exposure during cycle days 7–11 gave rise to a significant increase in the ratio of the length of follicular to luteal phases.Progesterone released during the early proliferative phase (days 2–6) exerted little, if any, effect on the appearance of the endometrium. However, the same dose of P released during the late proliferative phase (days 7 – 11) significantly diminished the number of glandular mitoses, the height of the glandular epithelium, reduced pseudostratification and the number of plasmolemmal vesicles, but did not induce any subnuclear vacuolation, predecidual reaction or leucocytic infiltration.It is suggested that systematic studies involving the exposure to progestogens in normally menstruating women during cycle days 7 to 11 will provide a reliable and practical method for the comparative assessment of the potency profile of individual progestogens in women.     

Synthesis and antimicrobial properties of new cepham and penam derivatives with the carboxylic group in the “wrong” β-configuration

The cepham (9) and the penam (10) derivatives, with their carboxylic group in the “wrong” β-configuration, which is opposite to that exhibited by the common penicillins, were synthesized and assayed in vitro for antimicrobial properties. Compounds 9 and 10, tested on Gram-positive and Gram-negative bacteria, both sensitive and resistant to β-lactamase inactivation, exhibited a very poor activity, directed only against non-β-lactamase-producing Gram-positive bacteria. One of the possible explanations for these results may be found in unfavourable steric and electronic effects deriving from the α-oriented methoxy group which is adjacent to the pharmacophoric carboxylic function.     

Cadmium metabolism in man

ABSTRACT Twenty-one high frequency solderers, who had been exposed to cadmium (Cd) from a solder for periods ranging from 1 month to 18 years (median 8 months; present time-weighted average 30 nmol/m³; particle size below 1μm) had Cd levels ranging from < 10 to 440 nmol/l in blood and from < 0·5 to 27 μmol/mol creatinine in urine. Individual workers showed considerable variations in blood Cd levels with time, but less variation in urine levels. There was a statistically significant (p < 0·001) increase of Cd in urine with increasing exposure time. Four gas solderers, who had been intermittently exposed for 8-20 years (median 17 years) had Cd levels ranging from 45 to 150 nmol/l and urine levels of from 2 to 20 μmol/mol creatinine. There was no correlation between Cd levels in blood and urine during exposure. After exposure had ceased there was a considerable decay of blood Cd in most subjects. The half-time in 11 people ranged from 25 to 146 days (median 41 days). After the decay blood levels reached a steady state. Concentrations in urine did not decrease, or did so only very slowly. There was a significant increase of levels in urine (p < 0·001) with increasing post-decay levels in blood. There was also a significant increase (0·01 < p < 0·05) of excretion of ß₂-microglobulin in urine (range 1·1-18 mg/mol creatinine, median 4·7 mg/mol creatinine) measured 11-15 months after exposure had ceased, with increasing Cd levels in urine. This may indicate an effect on renal tubular function even at kidney Cd loads corresponding to Cd levels in urine of the order of as little as 10 μmol/mol creatinine.     

Use of biocidal products (insect sprays and electro-vaporizer) in indoor areas – Exposure scenarios and exposure modeling

Five commercially available insect sprays were applied in a model room. Spraying was performed in accordance with the manufacturers’ instructions and in an overdosed manner in order to simulate worst-case conditions or an unforeseeable misuse. In addition, we examined electro-vaporizers. The Respicon™ aerosol monitoring system was applied to determine inhalation exposure. During normal spraying (10 seconds) and during the following 2–3 minutes, exposure concentrations ranged from 70 to 590 μg/m³ for the pyrethroids tetramethrin, d-phenothrin, cyfluthrin, bioallethrin, and the pyrethrins. Calculated inhalable doses were 2–16 μg. A concentration of approximately 850 μg chlorpyrifos/m³ (inhalable dose: approximately 20 μg) was determined when the “Contra insect fly spray” was applied. Highest exposure concentrations (1100–2100 μg/m³) were measured for piperonyl butoxide (PBO), corresponding to an inhalation intake of 30–60 μg. When simulating worst-case conditions, exposure concentrations of 200–3400 μg/m³ and inhalable doses of 10–210 μg were determined for the various active substances. Highest concentrations (4800–8000 μg/m³) were measured for PBO (inhalable: 290–480 μg).By applying the electro-vaporizer “Nexa Lotte” plug-in mosquito killer concentrations for d-allethrin were in the range of 5–12 μg/m³ and 0.5–2 μg/m³ for PBO while with the “Paral” plug-in mosquito killer concentrations of 0.4–5 μg/m³ for pyrethrins and 1–7 μg/m³ for PBO were measured.Potential dermal exposures were determined using exposure pads. Between 80 and 1000 μg active substance (tetramethrin, phenothrin, cyfluthrin, bioallethrin, pyrethrins, chlorpyrifos) were deposited on the clothing of the total body surface area of the spray user. Highest levels (up to 3000 μg) were determined for PBO. Worst-case uses of the sprays led to 5–9 times higher concentrations.Also a 2-hour stay nearby an operating electro-vaporizer led to a contamination of the clothing (total amounts on the whole body were 450 μg d-allethrin and 50 μg PBO for “Nexa Lotte” plug-in mosquito killer and 80 μg pyrethrins and 190 μg PBO for “Paral” plug-in mosquito killer).Human biomonitoring data revealed urine concentrations of the metabolite (E)-trans-chrysanthemum dicarboxylic acid ((E)-trans-CDCA) between 1.7 μg/l and 7.1 μg/l after 5 minutes of exposure to the different sprays. Also the use of electro-vaporizers led to (E)-trans-CDCA concentrations in the urine in the range of 1.0 μg/l to 6.2 μg/l (1–3 hours exposure period).The exposure data presented can be used for performing human risk assessment when these biocidal products were applied indoors.The airborne concentrations of the non-volatile active chemical compounds could be predicted from first principles using a deterministic exposure model (SprayExpo).     

Long-term consumption of a carbohydrate-restricted diet does not induce deleterious metabolic effects

Carbohydrate (CHO)-restricted diets have been recommended for weight loss and to prevent obesity, but their long-term effects have not been fully elucidated. This study was designed to evaluate the effect of long-term (>1 year) consumption of a low-CHO high-fat diet (“The optimal diet,” developed by Dr Kwaśniewski referenced herein) on lipid profile, glycemic control, and cardiovascular disease risk factors in healthy subjects. Of 31 “optimal” dieters enrolled in the study (17 women and 14 men, aged 51.7 ± 16.6 years), 22 declared adherence to the diet for more than 3 years. Average energy intake and principal nutrients consumed were assessed from 6-day dietary records provided by the participants. In most dieters, concentrations of β-hydroxybutyrate, free fatty acids, total cholesterol, and low-density lipoprotein cholesterol exceeded the upper limits of the reference ranges for nonstarved subjects. The metabolic profiles of most subjects were positive for several indicators, including relatively low concentrations of triacylglycerols, high levels of high-density lipoprotein cholesterol (HDL-C), and normal ratios of low-density lipoprotein cholesterol/HDL-C and total cholesterol/HDL-C. In most subjects, plasma concentrations of glucose, insulin, glucagon, cortisol, homocysteine, glycerol, and C-reactive protein were within reference ranges. Notably, in all but one subject, the homeostasis model assessment index of insulin resistance remained below the threshold for diagnosis of insulin resistance. These results indicate that long-term (>1 year) compliance with a low-CHO high-fat “optimal diet” does not induce deleterious metabolic effects and does not increase the risk for cardiovascular disease, as evidenced by maintenance of adequate glycemic control and relatively low values for conventional cardiovascular risk factors.     

Quantitative assessment of color vision impairment in workers exposed to toluene

Color vision was examined by the Lanthony-D-15 desaturated test in two groups of workers occupationally exposed to toluene and in a control group. Biological parameters of toluene exposure were analyzed: toluene in air and in venous blood, orthocresol, and hippuric acid in urine after workshift. The first exposed group, Group E₁, comprised 41 workers (toluene exposure ranged from 11.30 to 49.30 ppm), and the second exposed group, Group E₂, comprised 32 workers (toluene exposure ranged from 66.00 to 250.00 ppm). The nonexposed group, Group NE, comprised 83 subjects. Each group was divided into two subgroups; alcohol consumers and nonconsumers. Color vision loss was expressed as a color confusion index (CCI) and as age and alcohol intake-adjusted color confusion index (AACCI). Significantly higher values of CCI and AACCI (both P < 0.0001) in Group E₂ in comparison to Group NE, and significantly higher CCI (P < 0.0001) and AACCI (P < 0.05) values in Group E₂ in comparison to Group E₁ were established. The significant difference in CCI value between alcohol consumers and nonconsumers was established only in Group NE (P < 0.05). In Group NE significant correlation was found between CCI value as a dependent and age and alcohol intake as independent cofactors (R² = 0.45; P = 0.0000). In Group E₂ significant correlation was established between CCI as a dependent factor and age, toluene in air, and alcohol intake (R² = 0.72; P = 0.0001), or between CCI as dependent and age, toluene in blood and alcohol intake as independent cofactors (R² = 0.68; P = 0.0002). In Group E₁ significant correlation was established only between CCI and age (P < 0.005). In Group E_2, AACCI value significantly correlated with toluene in air (P < 0.0001), toluene in blood (r < 0.0005), orthocresol (P < 0.005) and hippuric acid (P < 0.005) in urine after workshift. There were no differences between smokers and nonsmokers in CCI values in the examined groups. Results of this study indicate that toluene in exposed workers can impair color vision. The role of alcohol intake and age influence on color vision loss cannot be ignored in such workers. Am. J. Ind. Med. 33:297–304, 1998. © 1998 Wiley-Liss, Inc.     

Oxidative stress and labile plasmatic iron in anemic patients following blood therapy

AIM: To determine the plasmatic iron content and evaluate the oxidative stress (OS) markers in subjects receiving blood therapy.METHODS: Thirty-nine individuals with unspecified anemia receiving blood transfusions and 15 healthy subjects were included in the study. Anemic subjects were divided into three subgrouP: (1) those that received up to five blood transfusions (n = 14); (2) those that received from five to ten transfusions (n = 11); and (3) those that received more than ten transfusions (n = 14). Blood samples were collected by venous arm puncture and stored in tubes containing heparin. The plasma and cells were separated by centrifugation and subsequently used for analyses. Statistical analyses were performed using Kruskal-Wallis analysis of variance followed by Dunn’s multiple comparison tests when appropriate.RESULTS: The eletrophoretic hemoglobin profiles of the subjects included in this study indicated that no patients presented with hemoglobinopathy. Labile plasmatic iron, ferritin, protein carbonyl, thiobarbituric acid-reactive substances (TBARS) and dichlorofluorescein diacetate oxidation were significantly higher (P < 0.05), whereas total thiol levels were significantly lower (P < 0.05) in transfused subjects compared to controls. Additionally, the activity of catalase, superoxide dismutase and glutathione peroxidase were significantly lower in the transfused subjects (P < 0.05). Antioxidant enzyme activities and total thiol levels were positively correlated (P < 0.05), and negatively correlated with the levels of protein carbonyl and TBARS (P < 0.05). In contrast, protein carbonyl and TBARS were positively correlated (P < 0.05). Altogether, these data confirm the involvement of OS in patients following therapy with repeated blood transfusions.CONCLUSION: Our data reveal that changes in OS markers are correlated with levels of labile plasmatic iron and ferritin and the number of transfusions.     

Chorionic gonadotropin prevents LRF-agonist-induced luteolysis in the human

The effects of exogenous and endogenous hCG on the luteolytic action of LRF-agonist, [D-Trp⁶, Pro⁹NEt]-LRF (LRF-Ag), were evaluated. In sequential studies, 4 normal cycling women treated with LRF-Ag (50 μg S.C.) on two consecutive days had premature decline of circulating levels of progesterone (P) and estradiol (E₂) with a shortened (p < 0.05) luteal phase (11.5 ± 1.2 days) when compared to the control cycle (15 ± 0.7 days). When intramuscular hCG was added to the LRF-Ag treatment in doses of 100 IU or 5000 IU daily for 7 days, the luteal function was prolonged (19 ±1.2 days, p < 0.05) with significant (p < 0.001) elevation of P and E₂ levels compared with the control cycle. Four women with early pregnancy, requesting therapeutic abortion, were given LRF-Ag (50 μ g or 500 μ g S.C.) on 2 consecutive days. None of the 4 women aborted and there was no change in the levels of _βhCG, P or E₂ over the course of a week. These results indicate that both exogenous or endogenous hCG can overcome the luteolytic effect of LRF-Ag within the dose and duration used. The possibility that a more prolonged administration of a larger dose of LRF-Ag may negate the luteotropic effect of hCG remains to be explored.     

Safety and Efficacy of Probiotic Supplementation in Reducing the Incidence of Infections and Modulating Inflammation in the Elderly with Feeding Tubes: A Pilot,Double-Blind,Placebo-Controlled Study, “IntegPRO”

A double-blind, placebo-controlled study was performed in a sample of geriatric patients treated with home enteral nutrition (HEN) to analyze the efficacy of a probiotic supplement Proxian^®, which contains Lactiplantibacillus plantarum LP01 (LMG P-21021), Lentilactobacillus buchneri Lb26 (DSM 16341), Bifidobacterium animalis subsp. lactis BS01 (LMG P-21384), and is enriched with zinc (Zn) and selenium (Se), in reducing the incidence of infections and modulating inflammation. Thirty-two subjects were enrolled (mean age 79.7 ± 10.3 years), 16 in the intervention group, 16 controls. They received Proxian^® or placebo for 60 days. Patients were assessed at baseline (t₀) and 60 (t₁) and 90 (t₂) days after the beginning. Infections were detected by information regarding their clinical manifestations and the incidence of antibiotic therapy. Levels of C-reactive protein (CRP) were measured to study inflammation. Information on bowel function, nutritional status and testimonials regarding the feasibility of administration of the product were collected. Differences between the two groups in number of infections (25% intervention group vs. 44% controls), antibiotic therapies (12% vs. 37%) and modulation of CRP levels (median CRP moved from 0.95 mg/L (t₀)_, to 0.6 (t₁) and 0.7 (t₂) in intervention group vs. 0.7 mg/L, 0.5 and 0.7 in controls) did not reach statistical significance. No significant changes in bowel function and nutritional status were found. Caregivers’ adherence was 100%. Results of this “IntegPRO” study showed that Proxian^® is potentially safe, easy to administer and promising for further studies but it appears not to change the incidence of infections or modulate inflammation in elderly treated with HEN. The utility of Proxian^® in reducing the incidence of infections and modulating inflammation in these subjects needs to be investigated by a larger multi-center clinical trial, and by using additional analyses on inflammatory markers and markers of infections.     

Comparing the spatial genetic structures of the Flavescence dorée phytoplasma and its leafhopper vector Scaphoideus titanus

The Nearctic leafhopper Scaphoideus titanus Ball is the vector of “Flavescence dorée” phytoplasma (FDp) in European vineyards. We studied the genetic diversity and structure of S. titanus populations in France and of the FDp they carried. A total of 621 S. titanus individuals, sampled in 24 FDp-infected and uninfected vineyards, were genotyped using seven polymorphic microsatellite loci. The mean observed heterozygosity in S. titanus populations was between 0.364 and 0.548. There was evidence of only a low level of population genetic differentiation (mean F_ST = 0.027) suggesting that there is long-distance gene flow between S. titanus populations. This may be a consequence of the high migration capacity of the vector associated with large effective population size and, at least in part, of passive dispersion over long distances by the transport of grapevine-planting material carrying eggs. For each insect, FDp was detected and typed by nested-PCR followed by RFLP and sequencing of a 674 bp fragment of the FDp map gene. Twelve of the 24 populations were found to be infected by FDp, with the percentage of infected individuals varying from 3% to 29%. FDp isolates were classified into two FDp genetic clusters (FD1 and FD2), which differed by 12–13 SNPs. FD1 genotypes were detected in the insect populations at two sites and the FD2 genotypes in the other ten populations. Both FD1 and FD2 genotypes were found to be transmitted by the insect. No significant relationship was found between the genetic structure of these French S. titanus populations and the distribution of the various FDp strain types they carried. Nevertheless, overall genetic differentiation between FDp-infected and healthy S. titanus “subsamples” was found to be significantly higher than zero. These results suggest that FDp-infected S. titanus individuals are more philopatric (disperse less) than healthy S. titanus.     

The effect of long-term supplementation with different dietary ω-6/ω-3 ratios on mineral content and ex vivo prostaglandin E2 release in bone of growing rabbits

BACKGROUND/OBJECTIVES

The aim of this research was to study the different long term effects of consumption of dietary oil sources with varying omega-6/omega-3 (ω-6/ω-3) polyunsaturated fatty acids (PUFAs) ratios on bone marrow fatty acid level, ex vivo prostaglandin E₂ (PGE₂) release, and mineral content of bone in rabbits.

MATERIALS/METHODS

For this purpose, weaning and female New Zealand white rabbits were purchased and randomly divided into five groups and offered ad libitum diets containing 70 g/kg of added oil for 100 days. The dietary lipid treatments were formulated to provide the following ratios of ω-6/ω-3 fatty acids: 8.68 soy bean oil (SBO control), 21.75 sesame oil (SO), 0.39 fish oil (FO), 0.63 algae oil (DHA), and 0.68 algae oils (DHA/ARA). DHA and ARA are two types of marine microalgae of the genus Crypthecodinium cohnii.

RESULTS

The dietary treatments had significant effects on the bone marrow fatty acids of rabbits. Rabbits fed the FO diet, containing the highest ω-3 PUFA concentration, and those fed the SBO diet showed the highest ω-6 PUFA. On the other hand, a positive correlation was observed between Ex vivo PGE₂ level and the ω-6/ω-3 dietary ratio. Significant effects of dietary treatment on femur Ca, P, Mg, and Zn contents were observed in both genders.

CONCLUSIONS

Findings of the current study clearly demonstrated that dietary PUFA, particularly ω-6/ω-3 and ARA/EPA ratios are important factors in determining bone marrow fatty acid profile, and this in turn determines the capacity of bone for synthesis of PGE₂, thereby reducing bone resorption and improving bone mass during growth.