Changes in the Level of Endogenous Leptin,FSH, 17β‐Oestradiol and Metabolites during Lupin‐induced Increase in Ovulation Rate in Ewes

The aim of the study was to determine the changes in the plasma concentration of leptin during lupin feeding‐induced increase in the ovulation rate (OR) in ewes. Additionally, alterations in the plasma level of glycogenic amino acids and glucose (as the factors influencing leptin secretion) and the levels of follicle‐stimulating hormone (FSH) and 17β‐oestradiol (E‐2) (as the hormones regulated by leptin and engaged in recruitment, selection and development of ovulatory follicles) were analysed. Ninety‐six female Polish Lowland Sheep were used. All ewes were cyclic and synchronized with PGF_2α. The ewes were divided into two groups: control (n = 48), fed only with hay, and experimental (n = 48), received additionally lupin (Lupinus angustifolius) grain as a high‐protein and a high‐energy supplement. They were given lupin from the second to 13th day of the oestrous cycle at increasing doses (150–750 g/day per ewe). On the 11th day of cycle blood samples for analysis of hormones, amino acids and total glucose concentration, were collected from the jugular vein. OR was determined by laparoscopy of ovaries on the sixth day of the following oestrous cycle. Mean OR of ewes supplemented with lupin grain (1.687 ± 0.463) was 30.67% higher than that of control (1.291 ± 0.454). In spite of the unchanged body mass, a significant increase (P ≤ 0.05) in mean concentration of plasma leptin in the experimental ewes [2.17 ± 0.15 ng/ml human equivalent (HE)] was found in comparison with control (1.42 ± 0.12 ng/ml HE). A significantly (P ≤ 0.05) higher plasma FSH level in the ewes fed lupin (105.21 ± 5.87 ng/ml) compared with those fed hay (67.88 ± 6.03 ng/ml) was also found. However, plasma level of E‐2 decreased after lupin feeding. Moreover, in the ewes fed lupin the plasma concentrations of glucose and nine glycogenic amino acids (Gly, Ala, Val, Met, Leu, Ile, Tyr, Phe and Arg) were increased. It can be concluded that lupin feeding exerts the stimulatory effect on the OR in Polish Lowland Sheep. The increase in OR is connected with significantly higher plasma leptin level and coincident with rise in FSH, glycogenic amino acids and glucose concentration. In contrast, the level of plasma E‐2 was significantly decreased in lupin‐fed ewes.     

Progesterone Profiles of Fat‐Tailed Tuj Ewes Following Tactile and Visual Separation of Rams at the Beginning of the Breeding Season

The aim of this study was to determine the level and duration of progesterone secretion during the sequential oestrous cycles in fat‐tailed Tuj ewes following tactile and visual separation of rams during the breeding season. For these purposes, rams were separated from the main flock for 50 days starting from the beginning of the breeding season and 21 ewes were randomly selected for the current experiment. In order to assess luteal activity and the length of oestrous cycles, the ewes were blood‐sampled thrice or twice weekly for the measurement of progesterone in plasma. The data showed that 3 ± 0.2 oestrous cycles were observed in this time window, and the first oestrous cycle observed was shorter (P < 0.05) than the following cycles (mean ± SEM, 14.6 ± 0.82, 16.5 ± 0.48 and 17.0 ± 0.54 days, respectively, for cycles 1, 2 and 3). Progesterone production was significantly lower in the first oestrous cycle compared with the second and third cycles on days 9, 12 and 14 of the cycles. The follicular periods of ewes showed four accumulations (maximum values using a 3‐day moving average technique) throughout the study and the percentages of ewes at each accumulation (peak day ± 1 day) were 50, 35, 65 and 80% for the first, second, third and fourth accumulations, respectively (P < 0.05). It was concluded that progesterone production was lower and the oestrous cycles were shorter during the first oestrous cycle and that tactile and visual separation of rams at the beginning of the breeding season might enhance the synchronizing effect towards the end of the breeding season in the fat‐tailed Tuj ewes.     

Pregnancy‐Specific Protein B (bPSPB) and Progesterone Monitoring of Post‐Partum Dairy Cows with Placental Retention

The relationship between placental retention, progesterone and pregnancy‐specific protein B (bPSPB) was determined in 60 calving Holstein cows. The cows were divided into two groups with placental retention (WPR, n=16) and no placental retention (NPR, n=44). Every 4 days, until 60 days post‐partum, blood samples were taken and the uteri were checked using ultrasonography. The puerperal characteristics of NPR and WPR were as follows: mean days of abnormal vaginal discharge: 20.2 ± 5 versus 35.6 ± 7 (P < 0.01); mean intervals to uterine involution: 21.4 ± 3.7 versus 27.6 ± 7.6 days (P < 0.01); rate of endometritis: 25 versus 100 % (P < 0.01). The mean numbers of oestrus cycles per cow were 1.75 ± 0.5 versus 0.85 ± 0.9 (P < 0.05) and the mean durations of the first oestrus cycle were 18 ± 3.5 versus 16 ± 2.1 days (P > 0.05). The mean intervals to first ovulation were 21.5 ± 8.4 versus 35 ± 19 days (P < 0.01). bPSPB blood concentrations were higher in the WPR group at calving with 955 ± 170 versus 750 ± 205 ng/ml (P < 0.01) and also during the first 32 days post‐partum with 173.68 ± 47.3 versus 131.0 ± 29.2 ng/ml (P < 0.01). The mean bPSPB half‐life was similar in the two groups: 6.9 ± 2.5 versus 6.5 ± 2.1 days (P > 0.05). In conclusion, it was found that placental retention was associated with a higher rate of endometritis, a lower number of cycles, longer interval to first post‐partum ovulation and higher concentration of bPSPB at calving and during the post‐partum period. The positive relationship between bPSPB concentrations and calf birth weight and their association with post‐partum pathological events may be useful in monitoring animals presenting high concentrations at calving.     

Conversion from twice‐daily to once‐daily tacrolimus formulation in pediatric liver transplant recipients – a long‐term prospective study

To assess the safety and efficacy of conversion from twice‐daily tacrolimus to once‐daily tacrolimus in pediatric liver transplant recipients. Conversion from twice‐daily to once‐daily tacrolimus was made in stable pediatric liver transplant recipients. Doses and serum levels of tacrolimus, liver, and renal function were recorded on the day before the conversion and at days 5, 30, 90, and 180 postconversion. Patients were controlled every 2–3 months thereafter. Fifty‐five patients were enrolled in the study. The mean age at conversion was 10.2 ± 3.6 years. The mean tacrolimus trough level was 4.7 ± 1.9 ng/dl preconversion, followed by a significant decline to 4.2 ± 1.7 30 days after the switch (P < 0.004). Mean daily tacrolimus dose was 0.09 ± 0.046 mg/Kg preconversion with a significant increase to 0.11 ± 0.060 3 months postconversion (P < 0.001). Fifteen patients with calculated glomerular filtration rate between 60 to 80 ml/min/m² preconversion showed a significant improvement one and 3 years after the switch (73 ± 4.1, 83 ± 4.3 and 90.3 ± 7.3 ml/min/m², respectively (P < 0.001). The mean follow‐up was 5.2 ± 2.4 years. Conversion to once‐daily tacrolimus is safe and effective in a cohort of stable pediatric liver transplant patients.     

Plasma levels of endothelin-1 in patients with the hepatorenal syndrome after successful liver transplantation

The hepatorenal syndrome (HRS) is characterized by renal vasoconstriction leading to deterioration of renal function in patients with liver disease. A possible role of endothelin-1 (ET-1) in the pathogenesis of HRS has been suggested, but a correlation between ET-1 plasma levels and the development of HRS as well as the recovery from HRS following OLT has not been shown yet. We performed longitudinal measurements of ET-1 plasma levels in four groups of patients, 5 patients with HRS before and after orthotopic liver transplantation (OLT), 10 patients without HRS undergoing OLT, 20 patients with chronic renal failure but without liver disease, and 12 healthy controls. Before OLT, plasma levels of ET-1 were higher in patients with HRS (19.5 ± 8.6 ng/l, P < 0.001; n = 5) compared to patients without HRS (4.9 ± 1.1 ng/l; n = 10), normals (1.2 ± 0.18 ng/l; n = 12), and patients with chronic renal failure (2.4 ± 0.4 ng/l; n = 20). Patients with HRS compared to patients without HRS had higher levels for creatinine (2.42 ± 0.6 vs. 0.89 ± 0.05 mg/dl, P < 0.05), creatinine clearance (107 ± 9 ml/min vs. 44.6 ± 5.5 ml/min, P < 0.001), and bilirubin (11.4 ± 3.8 vs. 3.7 ± 1 mg/dl, P < 0.05) before OLT. Within one week after OLT, there was a rapid decrease in ET-1 levels in patients with HRS while creatinine and bilirubin levels decreased slower. Regression analysis revealed a weak correlation between serum creatinine and ET-1 (r = 0.192, P = 0.04) and a significant correlation between serum bilirubin and ET-1 (r = 0.395, P < 0.001). The means of the ET-1 levels decreases rapidly with improvement of liver function after OLT. Levels of ET-1 correlate with excretory liver function assessed by bilirubin. The fall in ET-1 levels preceding improvement of renal function further strengthens the concept of ET-1 being a causative factor in HRS. Received: 22 July 1999/Revised: 28 January 2000/Accepted: 11 May 2000     

A randomized cross‐over comparison of short‐term exposure of once‐daily extended release tacrolimus and twice‐daily tacrolimus on renal function in healthy volunteers

Calcineurin inhibitor nephrotoxicity remains an issue for transplant recipients. The pharmacokinetic profile (PK) of the once‐daily tacrolimus extended release (Tac‐ER) includes equivalent exposure [AUC_(0–24 h)] but lower C_max versus twice‐daily tacrolimus immediate release (Tac‐IR). We hypothesized that the unique PK profiles would result in pharmacodynamic differences in renal function. Nineteen healthy male subjects were allocated to once‐daily Tac‐ER and twice‐daily Tac‐IR in a prospective, randomized, two period, cross‐over study. Tacrolimus was titrated to achieve trough levels of 8–12 ng/ml. Twenty four hours ERPF and GFR estimated by para‐aminohippurate and sinistrin clearance were performed at baseline and at the end of each 10‐day dosing period. Mean Tac C₀ was 11.0 ± 2.2 and 11.3 ± 1.8 ng/ml for Tac‐ER and Tac‐IR, respectively. The mean Effective 24 h renal plasma flow (ERPF) was significantly higher with Tac‐ER compared with Tac‐IR (658 ± 127 vs. 610 ± 93 ml/min/1.73 m², P = 0.046). There was a trend to a greater mean GFR over 24 h for Tac‐ER at 114.5 ± 13.6 ml/min/1.73 m² compared with 108.9 ± 9.7 ml/min/1.73 m² for Tac‐IR, P = 0.116. Under controlled physiological conditions, ERPF was significantly improved with Tac‐ER compared with Tac‐IR, likely owing to the differing PKs of these tacrolimus preparations (ClinicalTrials.gov Identifier: NCT01681134).     

The oxalate level in ultrafiltrate fluid collected from a dialyzer is useful for estimating the plasma oxalate level in hemodialysis patients

Background Patients on chronic hemodialysis are likely to develop secondary hyperoxalemia. It is, however, difficult to measure plasma oxalate levels. To measure plasma oxalate levels, rapid plasma separation, deproteinization, and acidification are essential in preventing the formation of oxalate and the deposition of calcium oxalate within the test tube. The present study was undertaken to examine whether the oxalate level in dialyzer ultrafiltrate is potentially useful for estimating plasma oxalate levels. Methods In nine patients on chronic hemodialysis, the plasma, after deproteinization with a filter, and the ultrafiltrate from the dialyzer before hemodialysis were acidified to a pH level of less than 3, followed by the measurement of oxalate levels by ion chromatography. Also, oxalate levels were compared between acidified and non-acidified ultrafiltrates from the dialyzer. In the second part of the study, seven patients on chronic hemodialysis receiving erythropoietin therapy, in whom the ferritin level was more than 300 ng/ml and transferrin saturation was less than 25%, were intravenously administered ascorbic acid, 100 mg, three times a week, after each dialysis session to facilitate the utilization of stored iron. This treatment was continued until the serum ferritin level decreased to a level below 300 ng/ml (for 3 months, at a maximum). The oxalate level in the dialyzer ultrafiltrate after this treatment was compared with that before treatment. Results The mean ± SE oxalate level in the dialyzer ultrafiltrate was 45 ± 6 μmol/l, essentially equal to the plasma oxalate level (46 ± 7 μmol/l). The plasma oxalate level had a significant positive correlation with the dialyzer ultrafiltrate oxalate level (plasma oxalate level = 0.99 × dialyzer ultrafiltrate oxalate level + 1.5; r = 0.95; P < 0.0001). The oxalate level in the acidified ultrafiltrate (45 ± 6 μmol/l) did not differ significantly from that in the non-acidified ultrafiltrate (45 ± 6 μmol/l). The mean ± SE duration of ascorbic acid administration was 64 ± 13 days. The hemoglobin level remained unchanged at 9.6 ± 0.4 g/dl, whereas the serum iron level increased significantly, from 34 ± 2 μg/dl to 43 ± 4 μg/dl (P < 0.05), and serum ferritin levels decreased significantly, from 645 ± 219 ng/ml to 231 ± 30 ng/ml after the treatment (P < 0.05). The oxalate level in the acidified ultrafiltrate showed no significant change after ascorbic acid administration (31 ± 8 μmol/l vs 47 ± 7 μmol/l). Conclusions In patients on chronic hemodialysis, the oxalate level in acidified ultrafiltrate from the dialyzer was found to be useful for estimating the plasma level of non-protein-bound oxalate. When administering ascorbic acid to hemodialysis patients, the plasma oxalate level can be monitored using this method.     

Sotrastaurin single‐dose pharmacokinetics in de novo liver transplant recipients

Sotrastaurin is a protein kinase C inhibitor in development for prevention of rejection after liver transplantation. In a pharmacokinetic study, 13 de novo liver transplant recipients received 100 mg sotrastaurin once between days 1–3 and once between days 5–8 post‐transplant. Sotrastaurin absorption based on the area under the concentration‐time curve (AUC) of total drug in blood (3544 ± 1434 ng·h/ml) was similar to that of healthy subjects in a previous study (4531 ± 1650 ng·h/ml). However, the sotrastaurin binding protein, α1‐acid glycoprotein, was nominally higher in patients (1.07 ± 0.28 vs. 0.87 ± 0.16 g/l, P = 0.13) yielding a 60% lower AUC based on free drug versus that in healthy subjects (27 ± 13 vs. 62 ± 15 ng·h/ml, P < 0.0001). There was minor excretion of sotrastaurin in drained bile (1% of dose) consistent with the fact that sotrastaurin is extensively metabolized leaving little unchanged drug to excrete. In the first week after liver transplantation, sotrastaurin is bioavailable after oral administration. However, patients with elevated α1‐acid glycoprotein levels may have lower free drug concentrations. Whether a higher dose of sotrastaurin is needed to compensate for this in the short‐term after surgery will be addressed in future clinical trials.     

Efficacy,safety, and immunosuppressant adherence in stable liver transplant patients converted from a twice‐daily tacrolimus‐based regimen to once‐daily tacrolimus extended‐release formulation

The aim of this study was to determine the efficacy, safety, and immunosuppressant adherence in 125 stable liver transplant (LT) patients converted from twice‐daily tacrolimus (TAC BID) to once‐daily TAC (TAC OD). Tacrolimus trough levels, laboratory parameters, metabolic disorders, selected patient reported outcomes, and adverse events were assessed. Mean TAC trough level concentration was 6.1 ± 2.3 ng/ml at study entry, decreased to 5.5 ± 2.1 ng/ml (P = 0.016) and 5.5 ± 2.2 ng/ml (P = 0.019) after 1 and 2 weeks, respectively, and tended to equal the baseline value during further follow‐up. At week 1, TAC concentrations were lower in 62.4% of patients and higher in 36.0% when compared with baseline. Renal and cardiovascular risk factors remained stable and no rejection episodes occurred over 12 months. Adverse events were consistent with the safety profile known from previous studies with TAC BID. Nonadherence measured by the “Basel Assessment of Adherence Scale to Immunosuppressives” was evident in 66.4% at study entry and decreased to 30.9% postconversion (P < 0.0001). Prevalence of nonadherence at baseline was significantly higher in patients converted >2 years after LT and in those ≤60 years of age. Conversion to TAC OD is safe, enhances immunosuppressant adherence and should be accompanied by a close TAC level monitoring during the initial period.     

Obstruction of St. Jude Medical Valves in the Aortic Position: Plasma Transforming Growth Factor Type Beta 1 in Patients With Pannus Overgrowth

The study investigated the hypothesis that plasma transforming growth factor type beta 1 (TGF‐β1) initiated pannus overgrowth in cases with aortic prosthetic valve dysfunction (PVD). Patients with obstruction of an aortic St. Jude Medical valve in 26 cases (PVD group) and without obstruction in 48 cases (control group) were studied. Plasma TGF‐β1, the intensity of the prothrombin time–international normalized ratio (PT‐INR), and the interruption of an oral anticoagulant medicine were conducted. Plasma TGF‐β1 levels in the PVD group (87.7 ± 29.2 ng/mL) were significantly higher (P < 0.05) than in the control group (73.7 ± 25.2 ng/mL). The interruption of an oral anticoagulant medicine in 54% of the PVD group versus 12% of the control group was identified (P < 0.001). The mean value of the PT‐INR in the PVD group (1.75 ± 0.30) and control group (1.75 ± 0.30) was not significantly different (P = 0.82). In conclusion, elevated levels of plasma TGF‐β1 may play a role in pannus overgrowth.     

Effects of hPTH(1‐34) Infusion on Circulating Serum Phosphate, 1,25‐Dihydroxyvitamin D,and FGF23 Levels in Healthy Men

Fibroblast growth factor 23 (FGF23) promotes phosphaturia and suppresses 1,25‐dihydroxyvitamin D [1,25(OH)₂D] production. PTH also promotes phosphaturia, but, in contrast, stimulates 1,25(OH)₂D production. The relationship between FGF23 and PTH is unclear, and the acute effect of pharmacologically dosed PTH on FGF23 secretion is unknown. Twenty healthy men were infused with human PTH(1‐34) [hPTH(1‐34)] at 44 ng/kg/h for 24 h. Compared with baseline, FGF23, 1,25(OH)₂D, ionized calcium (iCa), and serum N‐telopeptide (NTX) increased significantly over the 18‐h hPTH(1‐34) infusion (p < 0.0001), whereas serum phosphate (PO₄) transiently increased and then returned to baseline. FGF23 increased from 35 ± 10 pg/ml at baseline to 53 ± 20 pg/ml at 18 h (p = 0.0002); 1,25(OH)₂D increased from 36 ± 16 pg/ml at baseline to 80 ± 33 pg/ml at 18 h (p < 0.0001); iCa increased from 1.23 ± 0.03 mM at baseline to 1.46 ± 0.05 mM at hour 18 (p < 0.0001); and NTX increased from 17 ± 4 nM BCE at baseline to 28 ± 8 nM BCE at peak (p < 0.0001). PO₄ was 3.3 ± 0.6 mg/dl at baseline, transiently rose to 3.7 ± 0.4 mg/dl at hour 6 (p = 0.016), and then returned to 3.4 ± 0.5 mg/dl at hour 12 (p = 0.651). hPTH(1‐34) infusion increases endogenous 1,25(OH)₂D and FGF23 within 18 h in healthy men. Whereas it is possible that the rise in PO₄ contributed to the observed increase in FGF23, the increase in 1,25(OH)₂D was more substantial and longer sustained than the change in serum phosphate. Given prior data that suggest that neither PTH nor calcium stimulate FGF23 secretion, these data support the assertion that 1,25(OH)₂D is a potent physiologic stimulator of FGF23 secretion.     

Induction of Functional Differentiation and Growth Inhibition In Vitro of Canine Osteosarcoma by 22-Oxacalcitriol,Calcitriol and All-trans Retinoic Acid

The effects of 22-oxacalcitriol (OCT), calcitriol and all-trans retinoic acid (ATRA) on the induction of functional differentiation and growth inhibition of the canine osteosarcoma cell line POS were investigated in vitro via bone differentiation markers and proliferation assays, respectively. The intracellular alkaline phosphatase (ALP) activity and the gamma-carboxyglutamic acid osteocalcin (GLA-OC) and procollagen type I C peptide (PIP) production were used as markers of differentiation. Treatment with 10⁻⁸ m concentrations of all drugs for 72 h significantly inhibited growth (P < 0.0001) and increased ALP activity and GLA-OC and PIP production in POS. OCT, calcitriol and ATRA significantly increased the: ALP activity from 1.58 ± 0.14 μmol/min/mg protein (mean ± SD; control) to 2.50 ± 0.09 (P < 0.0001), 2.30 ± 0.14 (P < 0.0001) and 2.00 ± 0.14 (P = 0.0008), respectively; GLA-OC production from 0.71 ± 0.01 ng/ml (control) to 2.87 ± 0.01 (P < 0.0001), 2.87 ± 0.11 (P < 0.0001) and 1.36 ± 0.06 (P < 0.0001), respectively; and PIP production from 433.91 ± 23.29 ng/ml (control) to 536.54 ± 15.46 (P = 0.0002), 497.06 ± 1.99 (P = 0.0028) and 481.66 ± 0.01 (P = 0.0104), respectively. This study demonstrated that treatment with these drugs induced a phenotypic maturation of POS cells into cells that exhibit the properties of functionally mature bone cells with parallel growth inhibition. The effects of these drugs on functional differentiation may be useful to complement the progression of a normal osteogenic differentiation process in the sarcoma.     

Effects of Abdominal Surgery on Thiobarbituric acid Reactive Substances and Plasma Anti‐oxidative Capacity in Dairy Cows

The present investigation was undertaken to study the effects of surgical stress on thiobarbituric acid reactive substances (TBARS) and ferric reducing ability of plasma (FRAP) in dairy cows. Ten Holstein–Friesian dairy cows, mean age 5.26 years, admitted for treatment of left abomasal displacement (omentopexy), were used in our study. Blood samples were collected from the jugular vein prior to surgery, immediately and then 15, 30, 60, 90 min, and 2, 5, 10, and 24 h after reposition of abomasum. Surgical stress resulted in a significant increase in plasma cortisol concentrations (P < 0.001) with the highest mean levels 15 min after surgery (32.4 ± 18.1 μg/l) as well as in serum levels of glucose (P < 0.001) and non‐esterified fatty acids (P < 0.001). Similarly, surgery transiently enhanced the plasma levels of TBARS (P < 0.001). The highest TBARS plasma levels (0.91 ± 0.18 μmol/l) were recognized 60 min after reposition of abomasums. There were no significant differences in FRAP in dairy cows before and after reposition of abomasum. In conclusion, these data indicate that stress reaction caused by surgical correction of left displacement of abomasum can be associated with higher production of TBARS.     

Acute Effects of Ethanol on Mineral Metabolism and Trabecular Bone in Sprague-Dawley Rats

In order to assess the effects of acute ethanol intoxication on bone, 45 female Sprague-Dawley rats were studied. Five rats were sacrificed at baseline. The remainder received either ethanol (2 g/kg of body weight) intraperitoneally or isotonic saline. Rats were sacrificed in groups of 10 (5 intoxicated and 5 placebo) at 1, 4, 8, and 24 hours after injection. At the time of sacrifice, a blood sample was obtained and the 4th vertebra was excised for histomorphometric analysis of undecalcified bone. Effect of ethanol was assessed by an analysis of variance test using a Scheffé procedure. In ethanol-treated rats we observed (mean ± SD, ethanol versus controls, maximum difference point, P value) a significant decrease in osteiod surface with osteoblasts (42.86 ± 15.61% versus 64.57 ± 6.24%, P < 0.05); osteoclast number (0.05 ± 0.02 n/mm² versus 0.17 ± 0.09 n/nm², P < 0.05), and osteocalcin (36.9 ± 2.21 ng/ml versus 45.8 ± 5.1 ng/ml, P < 0.05). Osteoclast surface was initially reduced (0.129 ± 0.09% versus 0.425 ± 0.26%, P < 0.01) but showed a subsequent increase (0.765 ± 0.24% versus 0.226 ± 0.17%, P < 0.01) attributable to alcohol. There was also a significant decrease in serum Ca (8.51 ± 0.23 mg/dl versus 9.10 ± 0.29 mg/dl, P < 0.01) and parathyroid hormone values (23.51 ± 5.72 pg/ml versus 76.39 ± 11.66 pg/ml, P < 0.001). We conclude that acute alcohol intoxication in rats induces early striking changes in bone histology and analytical parameters, not completely reversed after 24 hours. These data are consistent with a toxic effect induced by alcohol on bone. Received 30 May 1996 / 31 December 1996     

Serum bone GLA protein in streak gonad syndrome

Summary Osteoporosis is one of the most common complications of streak gonad syndrome (SGS), however its pathogenesis is still unclear. Bone Gla protein (BGP) has been found to be a serum marker of bone turnover in various metabolic disease states. In the present study serum BGP and alkaline phosphatase (AP) were measured in 13 osteoporotic patients with SGS and in 56 healthy women. Mean (±SD) serum BGP levels were normal (7.5±2.0 ng/ml) in seven patients who had been on estrogen-progestin replacement therapy and became significantly elevated (P<0.001) 2 and 3 months after discontinuation of the treatment (15.3±2.3 and 13.2±1.0 ng/ml, respectively). Mean (±SD) serum AP (207±65 U/l) showed significant increases (P<0.05) 2 months after withdrawal of hormonal substitution (287±74 U/l). Mean (±SD) serum BGP (15.4±3.5) and AP (287±49) levels were significantly higher (P<0.001 and <0.05, respectively) in six patients with SGS who had not been on hormonal substitution. These findings are consistent with those obtained in postmenopausal women suffering from “high remodelling osteoporosis” and suggest that bone turnover in osteoporotic patients with SGS is increased and the skeletal loss is a consequence of accelerated bone loss rather than decreased bone formation.     

Increased lung clearance of isoflurane shortens emergence in obesity: a prospective randomized‐controlled trial

Background: There is a concern that obesity may play a role in prolonging emergence from fat‐soluble inhalational anaesthetics. We hypothesized that increased pulmonary clearance of isoflurane will shorten immediate recovery from anaesthesia and post‐anaesthesia care unit (PACU) stay in obese patients. Methods: After Ethics Review Board approval, 44 ASA I–III patients with BMI>30 kg/m² undergoing elective gynaecological or urological surgery were randomized after completion of surgery to either an isocapnic hyperpnoea (IH) or a conventional recovery (C) group. The anaesthesia protocol included propofol, fentanyl, morphine, rocuronium and isoflurane in air/O₂. Groups were compared using unpaired t‐test and ANOVA. Results: Minute ventilation in the IH group before extubation was 22.6±2.7 vs. 6.3±1.8 l/min in the C group. Compared with C, the IH group had a shorter time to extubation (5.4±2.7 vs. 15.8±2.7 min, P<0.01), initiation of spontaneous ventilation (2.7±2.3 vs. 6.5±4.5 min, P<0.01), BIS recovery >75 (3.2±2.3 vs. 8.9±5.8 min, P<0.01), eye opening (4.6±2.9 vs. 13.6±7.1 min, P<0.01) and eligibility for leaving the operating room (7.1±2.9 vs. 19.9±11.9 min, P<0.01). There was no difference in time for eligibility for PACU discharge. Conclusion: Increasing alveolar ventilation enhances anaesthetic elimination and accelerates short‐term recovery in obese patients.     

Vitamin K<Subscript>2</Subscript> (menaquinone 4) reduces serum undercarboxylated osteocalcin level as early as 2 weeks in elderly women with established osteoporosis

 Twenty elderly osteoporotic women with vertebral fracture(s) were randomly allocated to two groups; women in the MK₄ group received calcium with menaquinone 4 (MK₄) at a dose of 45 mg/day for 2 weeks, and women in the control group received calcium alone for the same period. Serum intact osteocalcin (OC) and undercarboxylated osteocalcin (uc-OC) levels were measured by immunoradiometric assay and enzyme immunoassay, respectively, at baseline and on the 7th and 14th days following the start of the treatment. There were no differences in the baseline data including age, weight, phylloquinone, menaquinone 4, menaquinone 7, OC, and uc-OC levels between the MK₄ group and the control group. Administration of MK₄ significantly raised the MK₄ level from 0.20 ± 0.10 (mean ± SE) pg/ml to 15.09 ± 5.62 pg/ml (P < 0.04), and reduced serum uc-OC levels from 2.80 ± 0.93 ng/ml to 1.76 ± 0.56 ng/ml (P < 0.05) at the end of the study, respectively. No significant changes in these levels were observed in the control group. Serum OC levels were stable during the period in both groups. In this randomized prospective study, the MK₄ group shows a reduction in the serum uc-OC level within 2 weeks without any significant change in OC, suggesting that the uc-OC is changed to carboxylated OC. This early effect of MK₄ on bone metabolism may be estimated by the measurement of serum uc-OC in elderly osteoporotic women with vertebral fractures. Received: January 21, 2002 / Accepted: November 6, 2002 Offprint requests to: T. Miki     

Abnormal cell calcium concentrations in cultured bone cells obtained from femurs of obese and noninsulin-dependent diabetic rats

Summary Cytoplasmic free calcium concentration [Ca²⁺]i was quantified in cultured bone cells with osteoblastic characteristics. The cells were obtained from femurs of obese (fa/fa) Wistar-Kyoto rats, from nonobese, noninsulin-dependent diabetic (NIDD) Sprague Dawley rats, and from their appropriate controls. [Ca²⁺]i was also determined in bone cells obtained fromin vivo insulin-treated NIDD rats. Obese (Wistar Kyoto) rats had increased body weight (313±13 vs. 249±4 g;P<0.01), decreased femur weights (0.68±0.05 vs. 0.89±0.05 g;P<0.05), similar glucose levels (148±5 vs. 139±3 mg/dl), and higher plasma insulin levels (6.0±0.5 vs. 0.7±0.1 ng/ml;P<0.01) when compared with their nonobese [(fa/+); (+/+)] littermates. Nonobese, NIDD rats, compared with their appropriate controls (nondiabetic Sprague Dawley rats) had higher plasma glucose levels (235±32 vs. 145±3 mg/dl;P<0.01) but their plasma insulins, body weights, and femur weights were similar to controls (0.7±0.1 vs 0.6±0.1 ng/ml; 302±4 vs. 318±14 g; 0.97±0.4 vs. 0.98±0.04 g, respectively). Long-term (4 weeks) daily insulin treatment (2 u/100 g) of the NIDD rats increased their plasma insulin (1.9 ng/ml;P<0.05) and body weight (369±13 g;P<0.05) but did not change their plasma glucose levels (225±5 mg/dl), or femur weights (0.98±0.4 g). [Ca²⁺]i in bone cells derived from the femurs of obese animals was higher than in cells derived from their nonobese littermates (156±22 vs. 71±13 nM;P<0.01). In bone cells from NIDD rats, [Ca²⁺]i was lower compared with controls (146±22 vs. 229±19 nM;P<0.001). Insulin treatment of the diabetic animals augmented the decrease in [Ca²⁺]i in their bone cells (68±11 nM;P<0.005 compared with nontreated rats). These data reveal that [Ca²⁺]i in cultured bone cells from obese nondiabetic and nonobese NIDD rats differs from that of their controls. Compared with their controls, the changes in [Ca²⁺]i in bone cells from the NIDD and obese animals were in opposite directions. Whether the underlying mechanisms for the changes in cell [Ca²⁺]i in nondiabetic obese and nonobese NIDD animals differ, and whether the changes in [Ca²⁺]i observed in the cultured cells reflect thein vivo condition in bone cells of these animals are questions that await further investigations.     

Diagnostic accuracy of neutrophil‐derived circulating free DNA (cf‐DNA/NETs) for septic arthritis

The release of “neutrophil extracellular traps” (NETs) has been identified as a novel immune response in innate immunity. NETs are composed of neutrophil‐derived circulating free DNA (cf‐DNA) and neutrophil cytoplasm‐derived proteins such as proteases. In this study, we analyzed the putative diagnostic value of synovial cf‐DNA/NETs for identification of septic arthritis. Forty‐two patients with a joint effusion who had undergone arthrocentesis were included. From synovial fluid, cf‐DNA/NETs (j‐cf‐DNA) levels were directly quantified. Diagnostic value of j‐cf‐DNA was compared with white blood cells (WBC), synovial white blood cells (j‐WBC), C‐reactive protein (CRP), j‐IL‐6, j‐TNF alpha, j‐IL‐1 beta, and myeloperoxidase (j‐MPO). Sensitivity, specificity, positive and negative predictive value, as well as ROC‐curves for each parameter were calculated. Synovial fluid cf‐DNA/NETs values from patients with septic arthritis (3,286 ± 386 ng/ml, n = 9) were significantly increased compared to patients with noninfectious joint inflammation (1,040 ± 208 ng/ml, n = 17) or osteoarthritis (278 ± 34 ng/ml, n = 16, p < 0.01). In conjunction with j‐cf‐DNA, j‐IL‐6 and j‐IL‐1 beta were significantly elevated (p < 0.01), but WBC, CRP, and j‐WBC were not. At a cut‐off of 300 ng/ml, j‐cf‐DNA had a sensitivity of 0.89, a specificity of 1.0, a positive predictive value of 1.0, and a negative predictive value of 0.97. Receiver operation curves revealed largest areas under the curve for cf‐DNA/NETs (0.933) and j‐IL‐6 (0.951). cf‐DNA/NETs seem to be a valuable additional marker for the diagnosis of septic arthritis or periprosthetic infections. However, this result should be confirmed in a large clinical trial. © 2009 Orthopaedic Research Society. Published by Wiley Periodicals, Inc. J Orthop Res 27:1401–1407, 2009     

Patients with high‐bone‐mass phenotype owing to Lrp5‐T253I mutation have low plasma levels of serotonin

The Lrp5 gene is a major determinant of bone mass accrual. It has been demonstrated recently to achieve this function by hampering the synthesis of gut‐derived serotonin, which is a powerful inhibitor of bone formation. In this study we analyzed plasma serotonin levels in patients with a high‐bone‐mass (HBM) phenotype owing to gain‐of‐function mutation of Lrp5 (T253I). A total of 9 HBM patients were compared with 18 sex‐ and age‐matched controls. In HBM patients, the serotonin concentrations in platelet‐poor plasma were significantly lower than in the controls (mean ± SEM: 2.16 ± 0.28 ng/mL versus 3.51 ± 0.49 ng/mL, respectively, p < .05). Our data support the hypothesis that circulating serotonin levels mediate the increased bone mass resulting from gain‐of‐function mutations in Lrp5 in humans. © 2010 American Society for Bone and Mineral Research.