The Diagnostic Performance of Urine Flow Cytometer UF1000i for Urinary Tract Infections.

BACKGROUND: Urinary tract infections (UTIs) are one of the most common infectious diseases in clinic. Urine flow cytometry is receiving more and more attention due to its rapid forecast of UTIs. METHODS: The Urine Flow Cytometer UF1000i has a series of software programs to quantify bacteria (BACT) and white blood cells (WBC), and describe the scatter diagram of bacteria. The UTIs were predicted based on the cutoff values with the Receiver Operating Characteristic (ROC) curves of BACT and WBC counts. To evaluate the diagnostic performance of UF1000i for UTIs, the sensitivity and specificity of 889 urine samples were determined in comparison to the results of urine culture. Meanwhile the bacterial morphology indication of the UF1000i was evaluated in order to help doctors choose antibiotics. The angle of the scatter cloud with the x-axis was used to classify the infected bacteria as bacilli (< 30°) or cocci (≥ 30°). RESULTS: The best cutoff value of BACT counts for predicting UTIs was 119 per µL, and the sensitivity and specificity were 95.5% and 88.7%, respectively. While the best cutoff value of WBC counts was 81.5 per µL, and the sensitivity and specificity were 77.6% and 76.7%, respectively. In addition, the best cutoff values for females were 583 BACT per µL and 137.5 WBC per µL. They were much higher than for males (118 BACT per µL and 91 WBC per µL). The coincidence of the bacterial morphology information between the UF1000i software indication and the bacterial actual morphology identified by urine culture was 83% (bacilli) and 68% (cocci), respectively. CONCLUSIONS: Data demonstrated that the performance of BACT counts for UTIs is superior to WBC counts. In addition, the bacterial morphology could preliminarily be predicated by the scatter diagram. Since the urine flow cytometer UF1000i can provide the data of both BACT counts and the scatter diagram, the urine flow cytometry was regarded as a suitable method for screening UTIs. Moreover, it would be better to take gender into consideration when setting the best cutoff value for diagnosis of UTIs in clinic.

Hepatitis B virus x protein induces autophagy via activating death-associated protein kinase.

Hepatitis B virus x protein (HBX), a product of hepatitis B virus (HBV), is a multifunctional protein that regulates viral replication and various cellular functions. Recently, HBX has been shown to induce autophagy; however, the responsible mechanism is not fully known. In this study, we established stable HBX-expressing epithelial Chang cells as the platform to study how HBX induced autophagy. The results showed that the overexpression of HBX resulted in starvation-induced autophagy. HBX-induced autophagy was related to its ability to dephosphorylate/activate death-associated protein kinase (DAPK). The block of DAPK by its siRNA significantly counteracted HBX-mediated autophagy, confirming the positive role of DAPK in this process. HBX also induced Beclin 1, which functions at the downstream of the DAPK-mediated autophagy pathway. Although HBX could activate JNK, a kinase known to participate in autophagy in certain conditions, the change in JNK failed to influence HBX-induced autophagy. In conclusion, HBX induces autophagy via activating DAPK in a pathway related to Beclin 1, but not JNK. This new finding should help us to understand the role of autophagy in HBX-mediated pathogenesis and thus may provide targets for intervening HBX-related disorders.

High resolution computed tomography and classification of children with interstitial lung diseases.

High resolution computed tomography (HRCT) was used to classify children with interstitial lung diseases (ILD). Sixty children with ILD underwent HRCT in supine position under free-respiratory conditions during scanning. Children under 5 years old were sedated with chloral hydrate and the scanning scope was from the lung apex to the diaphragm. In children older than 5 years old, scans were obtained at three levels: aortic arch, tracheal carina, and 1 cm above the right diaphragm. Five infectious patients were followed up. Two experienced radiologists read the films blindly to observe the type and distribution of ground-glass opacities and bronchovascular bundle abnormalities. Bronchovascular bundles were thick in 49 patients, and were thick and stiff in 27 patients. Of the 41 infectious patients, 39 showed thickened bronchovascular bundles, and 26 showed thick and stiff bronchovascular bundles. Of the 19 non-infectious patients, bronchovascular bundles were thickened in 10 patients, and were thick and stiff in 1 patient. Forty-one patients showed lobular ground-glass opacity (32 infectious, 9 non-infectious). Twenty-seven patients showed both bronchovascular bundle abnormality and lobular ground-glass opacity (20 infectious, 7 non-infectious). Eighteen patients showed patchy or mosaic ground-glass opacity (16 infectious, 2 non-infectious). There were 4 cases of bronchiectasis. HRCT is the first non-invasive diagnostic method for children with ILD, and its different manifestations can be classified. In early manifestation, bronchovascular bundles were abnormal and complicated with lobular ground-glass opacity. Patchy ground-glass opacity was the most common manifestation, and appeared to be difficult to disappear. Bronchiectasis indicated that the disease is irretrievable.

Effect of low doses of dietary rare earth elements on growth performance of broilers.

The present study was designed to investigate effect of dietary rare earth elements (REE), including both organic and inorganic compounds, on growth performance of broilers. In experiment 1, a total of 180 male Ross broiler chicks were allocated to 72 pens with different assignment: four chicks per pen or individually. The following three treatment diets were applied: control, REE-chlorides at a dose of 40 mg/kg and REE-citrate at a dose of 70 mg/kg. Each treatment group had 24 pens containing both assignments (12 pens each). In experiment 2, a total of 72 male 3-day-old Ross broiler chicks were separated to four groups: control, REE-chlorides at a dose of 70 mg/kg and REE-citrate at doses of 70 mg/kg and 100 mg/kg. In experiment 1, dietary REE-citrate improved body weight gain during the overall period by 5.0% (p < 0.05) while the increase with REE-chloride was not significant. In experiment 2, growth effects (p < 0.05) were only found in the period from day 21 to slaughter with all REE forms, and feed conversion ratio was improved by 3.4% (p < 0.05) with REE-citrate. No significant effects of REE were found on chill weight, percentages of breast meat, thigh weight, drumstick weight and wing weight. Concentrations of La and Ce in the liver and muscles were very low, accounting for 0.11-0.76 and 0.02-0.30 mg/kg respectively. There was weak tendency for a dose-response relationship especially in the groups supplemented with REE-chlorides. The main blood serum biochemical parameters were not significantly affected by REE in the diets. The results suggest that dietary supplementation of low doses of REE-citrates might improve growth performance of broilers without affecting carcass composition and health of the broilers.

Effect of a seaweed extract on fatty acid accumulation and glycerol-3-phosphate dehydrogenase activity in 3T3-L1 adipocytes.

This study was to determine the effect of a seaweed Ascophyllum nodosum extract (SE) containing 220 mg g(-1) phlorotannins on differentiation and fatty acid accumulation in differentiating 3T3-L1 adipocytes. 3T3-L1 cells (2 x 10(4) mL(-1)) were seeded to 24-well plates and proliferated to reach confluence and then were treated with media containing 0, 12.5, 25, 50, 75 and 100 mug mL(-1) SE for 8 days. Dexamethasone, methyl-isobutylxanthine and insulin (DMI) were added to the media in the first 2 days to induce cell differentiation. On day 8 the adipocytes were harvested for measuring cellular fatty acid concentration and the activity of glycerol-3-phosphate dehydrogenase (GPDH). It was found that treatment with SE increased (P < 0.01, n = 6) cellular myristoleic acid (C14:1), palmitoleic acid (C16:1) and oleic acid (C18:1) and total monounsaturated fatty acids (MUFA) without significantly affecting the cell number and saturated fatty acid (SFA). Ratios of MUFA/SFA, C14:1/C14:0, C16:1/C16:0 and C18:1/C18:0 in cellular lipids increased (P < 0.05, n = 6) with the SE treatment in a dose dependent manner (P < 0.001). Treatment with 75 microg mL(-1) SE depressed (P < 0.05) cellular GPDH activity. The results indicate that the biological factors in the SE may be involved in differentiation and MUFA accumulation in adipocytes.

Effects of the inclusion of Moringa oleifera seed on rumen fermentation and methane production in a beef cattle diet using the rumen simulation technique (Rusitec).

Moringa oleifera seeds are currently being used as a livestock feed across tropical regions of the world due to its availability and palatability. However, limited knowledge exists on the effects of the raw seeds on ruminant metabolism. As such, the rumen stimulation technique was used to evaluate the effects of substituting increasing concentrations of ground Moringa seeds (0, 100, 200 and 400 g/kg concentrate dry matter (DM)) in the diet on rumen fermentation and methane production. Two identical, Rusitec apparatuses, each with eight fermenters were used with the first 8 days used for adaptation and days 9 to 16 used for measurements. Fermenters were fed a total mixed ration with Urochloa brizantha as the forage. Disappearance of DM, CP, NDF and ADF linearly decreased (P<0.01) with increasing concentrations of Moringa seeds in the diet. Total volatile fatty acid production and the acetate to propionate ratio were also linearly decreased (P<0.01). However, only the 400 g/kg (concentrate DM basis) treatment differed (P<0.01) from the control. Methane production (%), total microbial incorporation of 15N and total production of microbial N linearly decreased (P<0.01) as the inclusion of Moringa seeds increased. Though the inclusion of Moringa seeds in the diet decreased CH4 production, this arose from an unfavourable decrease in diet digestibility and rumen fermentation parameters.

Carotenoids and prostate cancer risk.

Chemoprevention is presumably one of most effective means to combat prostate cancer (PCa). Patients usually require more than a decade to develop a clinically significant Pca, therefore, an ideal target for chemoprevention. This review will focus on recent findings of a group of naturally occurring chemicals, carotenoids, for potential use in reducing PCa risk.

Buffering agent via insulin-mediated activation of PI3K/AKT signaling pathway to regulate lipid metabolism in lactating goats.

Ruminants are often fed a high-concentrate (HC) diet to meet lactating demands, yet long-term concentrate feeding induces subacute ruminal acidosis (SARA) and leads to a decrease in milk fat. Buffering agent could enhance the acid base buffer capacity and has been used to prevent ruminant rumen SARA and improve the content of milk fat. Therefore, we tested whether a buffering agent increases lipid anabolism in the livers of goats and influences of milk fat synthesis. Twelve Saanen-lactating goats were randomly assigned to two groups: one group received a HC diet (Concentrate: Forage=60:40, Control) and the other group received the same diet with a buffering agent added (10 g sodium butyrate, C(4)H(7)NaO(2); 10 g sodium bicarbonate, NaHCO(3); BG) over a 20-week experimental period. Overall, milk fat increase (4.25+/-0.08 vs. 3.24+/-0.10; P<0.05), and lipopolysaccharide levels in the jugular (1.82+/-0.14 vs. 3.76+/-0.33) and rumen fluid (23,340+/-134 vs. 42,550+/-136) decreased in the buffering agent group (P<0.05). Liver consumption and release of nonesterified fatty acid (NEFA) into the bloodstream increased (P<0.05). Phosphatidylinositol 3-kinase (PI3K), protein kinase B (AKT) and ribosomal protein S6 kinase (p70S6K) up-regulated significantly in the livers of the buffering agent group (P<0.05). It also up-regulated expression of the transcription factor sterol regulatory element binding protein-1c (SREBP-1c) and its downstream targets involved in fatty acid synthetic, including fatty acid synthetase (FAS), stearoyl-CoA desaturase (SCD-1) and acetyl-CoA carboxylase 1 (ACC1) (P<0.05). The BG diet increased insulin levels in blood (19.43+/-0.18 vs. 13.81+/-0.10, P<0.05), and insulin receptor was likewise elevated in the liver (P<0.05). Cumulatively, the BG diet increased plasma concentrations of NEFA by INS-PI3K/AKTSREBP-1c signaling pathway promoting their synthesis in the liver. The increased NEFA concentration in the blood during BG feeding may explain the up-regulated in the milk fat of lactating goats.

Buffering agent-induced lactose content increases via growth hormone-mediated activation of gluconeogenesis in lactating goats.

Dairy goats are often fed a high-concentrate (HC) diet to meet their lactation demands; however, long-term concentrate feeding is unhealthy and leads to milk yield and lactose content decreases. Therefore, we tested whether a buffering agent is able to increase the output of glucose in the liver and influence lactose synthesis. Eight lactating goats were randomly assigned to two groups: one group received a HC diet (Concentrate : Forage = 6:4, HG) and the other group received the same diet with a buffering agent added (0.2 % NaHCO(3), 0.1 % MgO, BG) over a 19-week experimental period. The total volatile fatty acids and lipopolysaccharide (LPS) declined in the rumen, which led the rumen pH to become stabile in the BG goats. The milk yield and lactose content increased. The alanine aminotransferase, aspartate transaminase, alkaline phosphatase, pro-inflammatory cytokines, LPS and lactate contents in the plasma significantly decreased, whereas the prolactin and growth hormone levels increased. The hepatic vein glucose content increased. In addition, pyruvate carboxylase (PC), phosphoenolpyruvate carboxykinase (PEPCK) and glucose-6-phosphatase (G6PC) expression in the liver was significantly up-regulated. In the mammary glands, the levels of glucose transporter type 1, 8, 12 as well as of sodium-glucose cotransporter 1 increased. Cumulative buffering agent treatment increased the blood concentrations of glucose via gluconeogenesis and promoted its synthesis in the liver. This treatment may contribute to the increase of the milk yield and lactose synthesis of lactating goats.

Effects of garlic and juniper berry essential oils on ruminal fermentation and on the site and extent of digestion in lactating cows.

The objective of this study was to evaluate the effects of feeding essential oils from garlic (GAR) and juniper berry (JUN), or monensin (MO) on feed intake, ruminal fermentation, the site and extent of digestion, microbial protein synthesis, milk production, and immune status in dairy cows. Four midlactating Holstein cows fitted with ruminal and duodenal cannulas were used in a 4 x 4 Latin square design with 21-d periods and 4 treatments: control (no additive), MO (330 mg/cow per d), GAR (5 g/cow per d), and JUN (2 g/cow per d). Cows were fed ad libitum a TMR consisting of 40% forage and 60% barley-based concentrate. Dry matter intake averaged 20.4 kg/d and was not affected by dietary additives. Total tract digestibilities of dry matter, organic matter, fiber, and starch were not affected by experimental treatments. However, ruminal digestibilities of dry matter and organic matter were higher (+13%) for GAR and JUN than for the control diet, mainly because of increased crude protein digestion in the rumen. Feeding GAR and JUN increased ruminal digestion of dietary protein by 11% as compared with the control. In contrast, ruminal digestion of dietary protein was reduced by 11% with MO as compared with the control. Milk fat content was lower for MO (2.68%) than for the GAR (3.46%), JUN (3.40%), and control (3.14%) diets. No effects of GAR, JUN, or MO were observed on milk production, ruminal microbial protein synthesis, ruminal pH, and ruminal concentrations of volatile fatty acids and ammonia N. The total and differential numbers of white blood cells as well as serum amyloid A and haptoglobin were not affected by the treatments, suggesting that additives had no effect on the immune status of cows. Results of this study indicate that supplementing dairy cows with GAR (5 g/d) and JUN (2 g/d) essential oils improved feed digestibility in the rumen, but possibly at the expense of a reduction in the flow of bypass protein to the small intestine. Feeding monensin could be beneficial in terms of increasing bypass protein from the rumen but did not improve feed digestion or milk production under the current experimental conditions.

[Association between behavioral problems and gastrointestinal disorders among children with autism spectrum disorder].

Objective: To investigate the relationship between gastrointestinal disorders (GID) and core symptoms or behavioral problems among the children with autism spectrum disorder (ASD) . Method: Totally 328 children with ASD and 202 normal controls were enrolled in this cross-sectional study from August 2013 to October 2016. The information about the gastrointestinal disorders, behavioral and emotional problems was collected by using questionnaires. Childhood Autism Rating Scale (CARS), Autism Behavior Checklist (ABC) were used to assess the core symptoms of the children with ASD. Neurodevelopmental status was evaluated with Gesell Developmental Scale (GDS). These variables were analyzed by using student's t-test and chi-square test. Result: The prevalence of GID was significantly higher in the children with ASD than in the normally developing children (49.4% (162/328) vs.25.7% (52/202), χ(2)=29.039, P=0.000), especially the symptoms of constipation (33.2% (109/328) vs. 13.9% (28/202)), diarrhea (9.5%(31/328) vs. 1.5% (3/202)), nausea and vomiting (9.5% (31/328) vs. 3.5% (7/202)), and foul defecation (16.5% (54/328) vs. 5.0% (10/202)) (all P<0.05). Among the ASD children, the prevalence of GID was similar between male and female (46.7% (133/285) vs. 46.5%(20/43), χ(2)=0.006, P=0.938), as well as among all age groups (χ(2)=1.907, P=0.862). There was no significant difference in scores of GDS in the ASD children with or without GID (all P>0.05). Compared with ASD children without GID (n=166), the ASD children with GID (n=162) got higher scores in the "Body and Object Use" of ABC scale ( (16.4±9.3) vs. (12.3±6.7) scores, t=2.258, P=0.028), and had more emotional problems (63.6% (103/162) vs. 49.4% (82/166), χ(2)=6.707, P=0.010). Moreover, the score of behavior problems questionnaire was higher in the ASD children with GID ( (35.3±16.8) vs. (16.1±13.6) scores, t=5.748, P=0.000). Conclusion: Children with ASD have higher risk of GID than the normal developing children. While the stereotyped behaviors, problem behaviors and emotional problems are severer in the ASD children with GID. Hence, it is important to provide comprehensive treatment and management for these groups of children.

Effect of conjugated linoleic acid type, treatment period, and dosage on differentiation of 3T3 cells.

This study was conducted to determine effect of CLA and linoleic acid (LA) on cell differentiation, cellular glycerol-3-phosphate dehydrogenase (GPDH) activity, and FA accumulation in differentiating 3T3-L1 cells (3 isomers x 3 treatment periods x 4 doses). The cells were cultured in 24-well plates for proliferation until confluence. Then they were treated with media containing 0, 10, 35, or 70 mg/L (0, 35, 125, or 250 mmol/L, respectively) of LA, cis9,trans11- or trans10,cis12-CLA during early (day 0-2), intermediate and late (day 3-8), or overall (day 0-8) differentiation periods. Dexamethasone, methyl-isobutylxanthine, and insulin were supplemented to the media only for the early period to induce the differentiation. On day 8 of postconfluence the cells were harvested for Oil Red O staining, analysis of GPDH activity, and determination of the FA Concentration. Cellular LA or CLA was found to accumulate in a dose-response manner, mainly during the intermediate/late period. Treatment with trans10,cis12-CLA lowered (P < 0.05) GPDH activity and the concentration of FA including palmitic acid (16:0) and palmitoleic acid (16:1), especially during the intermediate/late and overall periods, or whenever a high dose of 70 mg/L was applied. This also resulted in a higher (P < 0.05) ratio of saturated FA to monounsaturated FA. Treatment with LA or cis9,trans11-CLA lowered cellular FA only when they applied during the early period at a dose of 70 mg/L. The results demonstrated that the inhibitory effects of CLA on differentiation, GPDH activity, and FA accumulation of 3T3-L1 cells are dependent on the isomer type, treatment period, and dose.

Effects of various dietary lipid additives on lamb performance, carcass characteristics, adipose tissue fatty acid composition, and wool characteristics.

Tasco (Ascophyllum nodosum; TA) was compared to canola (CO), flax (FO), and safflower oils (SO) for effects on performance, carcass characteristics, and fatty acid profiles of adipose tissue in skirt muscle (SM), subcutaneous and perirenal adipose tissues, and wool production and quality characteristics of Canadian Arcott lambs. Fifty-six lambs were randomly assigned to dietary treatments (n = 14 per treatment). Diets consisted of a pelleted, barley-based finishing diet containing either TA, CO, FO, or SO (2% of dietary DM). Feed deliveries and orts were recorded daily. Lambs were weighed weekly and slaughtered once they reached ≥ 45 kg BW. Carcass characteristics, rumen pH, and liver weights were determined at slaughter. Wool yield was determined on mid-side patches of 100 cm2 shorn at d 0 and on the day before slaughter (d 105 or 140). Dye-bands were used to determine wool growth, micrometer and staple length. Adipose tissues and SM samples were taken at slaughter and analyzed for FA profiles. No effects were observed on intake, growth, or carcass characteristics. A greater (P = 0.02) staple strength of lambs fed CO was the only effect observed in wool. Flax oil increased total n-3 and decreased the n-6/n-3 ratio in tissue FA profiles (P < 0.001) in comparison to other diets. Tasco increased (P ≤ 0.001) SFA/PUFA in all tissues, whereas concentrations of CLA c-9, t-11 were greatest with SO in all tissues (P ≤ 0.02), compared to other diets. These results suggest Tasco supplementation did not improve the n-3/n-6 or SFA/PUFA ratios of lamb adipose tissues compared to other dietary lipid additives.

Effect of wheat dried distillers grains and enzyme supplementation on growth rates, feed conversion ratio and beef fatty acid profile in feedlot steers.

The objectives of this study were to determine: (1) the effect of wheat dried distillers grain with solubles (DDGS) inclusion, and (2) dietary feed enzyme (FE; Econase XT) supplementation in a finishing diet containing wheat DDGS on fatty acid profile of the pars costalis diaphragmatis muscle of beef cattle. A total of 160 crossbred yearling steers with initial BW of 495 ± 38 kg were blocked by BW and randomized into 16 pens (10 head/pen). The pens were randomly assigned to one of the four treatments: (1) control (CON; 10% barley silage and 90% barley grain-based concentrate, dry matter (DM) basis); (2) diet containing 30% wheat DDGS in place of barley grain without FE (WDG); (3) WDG diet supplemented with low FE (WDGL; 1 ml FE/kg DM); and (4) WDG diet supplemented with high FE (2 ml FE/kg DM). The pars costalis diaphragmatis muscle samples were collected from cattle at slaughter at the end of the finishing period (120 days) with a targeted live weight of 650 kg. No differences in organic matter intake, final BW and average daily gain were observed among treatments. However, steers fed WDG had greater (P<0.01) feed conversion ratio than those fed CON, and increasing FE application in wheat DDGS-based diets tended (P<0.10) to linearly decrease feed conversion ratio. In assessing the effects of including WDG diets without FE, concentration of total polyunsaturated fatty acids (PUFA) in muscle tended to be greater (P<0.10) for steers fed WDG than steers fed CON. In addition, inclusion of wheat DDGS into the diet increased (P<0.05) concentration of CLA and vaccenic acid (VA) in muscle and also resulted in a higher (P<0.05) ratio of n-6/n-3 PUFA compared with that from steers fed CON diet. Increasing FE application in wheat DDGS-based diets did not modify the concentrations of individual or total fatty acids. These results suggest that inclusion of wheat DDGS in finishing diets may improve fatty acid profile of beef muscle which could benefit human health.

Effect of grain type and processing index on growth performance, carcass quality, feeding behavior, and stress response of feedlot steers.

One hundred sixty crossbred steers (538 ± 36 kg BW) were used in an 84-d experiment with a randomized block design to study the effects of wheat or barley grain processed to 2 different indices on growth performance, feeding behavior, carcass characteristics, stress, and temperament of finishing beef cattle. Treatments were a wheat-based diet (88.4% of diet DM; WH) and a barley-based diet (89% of diet DM; BA), processed to an index of either 75% (HI) or 85% (LO) of their original volume weight. Cattle were allocated to 16 feedlot pens (10 animals per pen, 4 pens per treatment), 8 of which were equipped with the GrowSafe system for monitoring feeding behavior. Flight speed, hair, and saliva samples were collected on d 1, 28, 56, and 84 to determine temperament, acute, and chronic stress. All steers were slaughtered at the end of the experiment, and carcass quality was evaluated. Cattle fed WH had a lower (P < 0.05) meal length and frequency of visits per meal and tended (P = 0.10) to have a lower DMI, meal size, and feeding time than those fed BA. The LO processing index increased (P = 0.05) DMI and reduced (P < 0.05) the G:F and the percentage of saleable meat of the carcass compared to HI. There was a trend (P = 0.09) for a grain × processing index interaction, where cattle fed BA-LO had a lower incidence of severe liver abscesses compared with cattle fed other treatments. Cattle fed WH had greater hair cortisol concentrations (P = 0.01) and flight speed (P < 0.01) than those fed BA. There was a trend (P = 0.07) for a grain × processing index interaction, where heifers fed WH-LO had a lower salivary cortisol than those fed other treatments. Results suggest that a LO processing index had a negative effect on feed efficiency and carcass performance and that the WH diet caused a range of effects on feed intake and behavior indicative of steers with greater excitability and chronic stress.

Effect of some oral contraceptives on serum concentrations of sex hormone binding globulin and ceruloplasmin.

Serum SHBG and ceruloplasmin (CP) concentrations were measured in women throughout a cycle of treatment with four oral contraceptives. In women receiving 150 micrograms levonorgestrel (LNG) daily both SHBG and CP decreased. SHBG also decreased, but CP increased, in women receiving ethynyloestradiol (EE) 30 micrograms with LNG 150 micrograms. In women taking 35 micrograms EE with either 600 micrograms or 1000 micrograms norethisterone, increases in CP were similar but SHBG increased more with the lower dose. Serum concentrations had not returned to pretreatment levels by eight days after cessation of dosing. The findings are compared with similar results for women taking 30 micrograms or 50 micrograms EE or an EE,LNG triphasic formulation. Serum concentrations of the gestagens were also measured. Increases in these concentrations when the gestagen was administered with EE to levels higher than expected from administration of the gestagen alone cannot be explained by increased binding to SHBG but are more likely to be due to changes in their metabolism. Differences in the responses of ostensibly closely related proteins of hepatic origin such as SHBG and CP to the oral contraceptives demonstrate that neither can be extrapolated to other pharmacodynamic responses.

PIP: 4 groups, each of 6 healthy volunteer female subjects aged 18-40 years, participated in this study designed to determine the effect of some contraceptive steroids on 2 plasma proteins -- sex hormone binding globulin (SHBG) and ceruloplasmin (CP). The pills were taken daily for 20 days, starting within the 1st 5 days of the menstrual cycle. There was no significant difference between the groups in the pre-treatment values. In Group LNG150 (150 mcg levonorgestrel), SHBG concentrations decreased, the decrease being statistically significant both in actual concentration and percentage change within 2-3 days of starting treatment. As pill-taking continued, SHBG concentrations declined further, and by the end of the treatment period they were less than half of the pretreatment value. Recovery was slow. In group EE30, LNG150 (ethinylestradiol 30 mcg and levonorgestrel 150 mg), no significant change occurred in SHBG concentrations during treatment. In Group EE35, NET1000 (ethinylestradiol 35 mcg and norethisterone 1000 mcg), SHBG increased, the value on days 2-3 of treatment being significantly higher than pretreatment. A steady state appeared to have been reached by days 7-8 and for the remainder of the treatment period the increase varied from 180-200% of the pretreatment value. The return to pretreatment values was slow. Women in Group EE35, NET600 (ethinylestradiol 35 mcg and norethisterone 600 mcg) also showed a rise in SHBG concentrations, which was greater than that observed in Group EE35, NET1000. In Group LNG150, CP concentrations slowly decreased during pill-taking, but the decrease did not reach statistical significance until days 14-15 of treatment; at this time the concentrations were about 15% lower than pretreatment. Despite stopping treatment, concentrations continued to decrease and were more than 25% lower by day 8 posttreatment. In Group EE30, LNG150, CP concentrations were significantly higher than pretreatment by days 2-3 and values continued to increase to the day of treatment. Women in Groups EE35, NET 1000 and EE35, NET600 showed a similar pattern, but the maximum percentage increase was higher, about 100%, compared to 75% in Group EE30, LNG150. Differences in the responses of ostensibly closely related proteins of hepatic origin, i.e., SHBG and CP, to the OCs demonstrate that neither can be extrapolated to other pharmacodynamic responses.

Effect of Anordrin on serum levels of sex hormone binding globulin, caeruloplasmin and ovarian function.

Ten doses of 7.5 mg Anordrin were administered on alternate days from day 5 of the menstrual cycle to 20 women. Serum concentrations of both SHBG and CP were significantly increased after the fourth dose but the increases were minor compared to those produced by ethynyloestradiol. Two weeks after stopping Anordrin, serum concentrations of the proteins were still elevated in most women. Ovarian activity was studied in 8 women; serum oestradiol and progesterone concentrations indicated that ovulation occurred at the normal time in three women but was delayed for two weeks or more in the remaining five. The findings suggest that in humans, Anordrin acts as a weak oestrogen and may also have antioestrogenic activity.

A pharmacodynamic and pharmacokinetic study of the Chinese No. 1 pill.

A pharmacodynamic and pharmacokinetic study of the Chinese No. 1 pill, a combined oral contraceptive containing 35 micrograms ethynyloestradiol (EE) and 600 micrograms norethisterone (NET), was performed in 29 women over a period of six months. Blood samples for analysis were taken during a pretreatment cycle, the first and 6th treatment cycles and post-treatment. Minor changes in carbohydrate metabolism occurred and these were particularly noticeable when the incremental areas under the serum concentration-time curves for both glucose and insulin in response to a glucose tolerance test were calculated. No changes occurred in the serum glycosylated haemoglobin levels. The serum concentrations of all the lipids measured (total cholesterol, triglycerides, LDL-C, HDL-C and apolipoproteins AI, AII and B) were significantly increased on treatment as were levels of Factor X, SHBG and caeruloplasmin whereas antithrombin III decreased. In 38 of the 40 treatment cycles, ovulation was suppressed. In one cycle serum oestradiol and progesterone levels showed a typical ovulatory pattern and in another there was evidence of follicular activity without ovulation. Serum EE concentrations showed a similar pattern in both treatment cycles showing that co-administration of NET did not affect EE metabolism. Serum NET levels were higher in the 6th than in the first treatment cycles. On comparing pharmacodynamic and pharmacokinetic parameters, the only statistically significant correlations were between the percentage change in triglycerides and SHBG and serum NET, but not EE concentrations, and between apolipoproteins AI and serum EE.

A cross-over study of three oral contraceptives containing ethinyloestradiol and either desogestrel or levonorgestrel.

A randomised cross-over trial was performed to compare the pharmacodynamic actions of three low-dose oral contraceptives (OCs): Marvelon (150 micrograms desogestrel (DSG)+ 30 micrograms ethinyloestradiol (EE)), Mercilon (150 micrograms DSG + 20 micrograms EE) and Microgynon (150 micrograms levonorgestrel (LNG) + 30 micrograms EE). None of the OCs produced any significant changes in serum cholesterol, LDL-C and apoprotein B. Triglycerides were increased by the desogestrel OCs but not by Microgynon. The latter however increased the glucose and insulin responses to a glucose tolerance test whereas Marvelon and Mercilon had no effect. HDL-C increased with Marvelon, was unchanged with Mercilon and was decreased with Microgynon. Apoprotein AII was increased by all three OCs but only the DSG OCs increased apoprotein AI. All OCs produced similar increases in caeruloplasmin but the increase in SHBG was much greater with Marvelon and Mercilon than with Microgynon. Testosterone was reduced more with Microgynon than with the DSG OCs. Many of the changes reflect the strong anti-oestrogenic action of LNG on metabolic parameters compared to DSG. Except for the effect on HDL-C, there was little difference between Marvelon and Mercilon on metabolic parameters and this complements the findings from large-scale clinical trials of the two OCs. Mercilon, therefore provides a very satisfactory alternative to Marvelon.

PIP: 12 healthy volunteers attending the family planning clinic at Shanghai, First Maternity and Infant China, Hospital, enrolled in the study, Oral contraceptives (OCs) were prescribed: Marvelon (150 mcg of desogestrel--DSG), Mercilon (150 mcg of DSG), and Microgynon (150 mcg of levonorgestrel--LNG). The patients were divided into 6 groups of 2 persons each in a randomized cross-over study. OCs were taken on day 6 of the cycle up to day 21, then stopping for 7 days. Each OC was used for 3 months. During the pretreatment cycle between days 6 and 9 of the follicular phase and 21 and 22 of the luteal phase a blood sample was taken after fasting for determination of lipids, sex hormone binding globulin (SHGB), ceruloplasmin, and testosterone. After glucose loading, significant increases of glucose and insulin occurred at 1, 2, and 3 hours during treatment with Microgynon only. The ratio for total areas of insulin to glucose did not change significantly nor did glycosilated hemoglobin A1 levels. Serum triglyceride concentrations increased significantly for both Marvelon (27%-43%) and Mercilon (29-40%). Serum high density lipoprotein (HDL) cholesterol concentrations were significantly elevated with Marvelon but less so with Mercilon, while HDL-C decreased significantly with Microgynon. The serum low density lipoprotein (LDL) cholesterol changes were not significant, but LDL-C concentrations declined with DSG formulations and increased with Microgynon. Apoprotein A1 and A2 increased significantly for both Marvelon and Mercilon. Apoprotein A2 increased with Microgynon. Serum SHBG increased markedly with Marvelon (335-380%). Serum testosterone concentrations decreased significantly (33.2-40.4% with Microgynon) and so did ceruloplasmin values. The antiestrogenic effect of strong LNG in Microgynon produced significant metabolic changes. The effect of 30 mcg EE in Marvelon and 20 mcg EE of Mercilon was equal.