Metagenomic characterization of the equine endometrial microbiome during anestrus.

The equine uterus is highly interrogated during estrus prior to breeding and establishing pregnancy. Many studies in mares have been performed during estrus under the influence of high estrogen concentrations, including the equine estrual microbiome. To date, it is unknown how the uterine microbiome of the mare is influenced by cyclicity; while, the equine vaginal microbiome is stable throughout the estrous cycle. We hypothesized that differences would exist between the equine endometrial microbiome of mares in estrus and anestrus. The aim of this study was two-fold: to characterize the resident endometrial microbiome of healthy mares during anestrus and to compare this with estrus. Double-guarded endometrial swabs were taken from healthy mares during estrus (n = 16) and in the following non-breeding season during anestrus (n = 8). Microbial population was identified using 16S rRNA sequencing. Our results suggest that the equine uterine microbiome in estrus has a low diversity and low richness, while during anestrus, a higher diversity and higher richness were seen compared to estrus. Despite this difference, both the estrus and anestrus endometrial microbiome were dominated by Proteobacteria, Firmicutes, and Bacteroidota. The composition of the microbial community between anestrus and estrus was significantly different. This may be explained by the difference in the composition of the endometrial immune milieu based on the stage of the cycle. Further research investigating the function of the equine endometrial microbiome and dynamics changes within the uterine environment is required.

Effect of Sampling Method on Detection of the Equine Uterine Microbiome during Estrus.

Bacterial endometritis is among the most common causes of subfertility in mares. It has a major economic impact on the equine breeding industry. The sensitivity of detecting uterine microbes using culture-based methods, irrespective of the sample collection method, double-guarded endometrial swab, endometrial biopsy, or uterine low-volume lavage (LVL), is low. Therefore, equine bacterial endometritis often goes undiagnosed. Sixteen individual mares were enrolled, and an endometrial sample was obtained using each method from all mares. After trimming, quality control and decontamination, 3824 amplicon sequence variants were detected in the dataset. We found using 16S rRNA sequencing that the equine uterus harbors a distinct resident microbiome during estrus. All three sampling methods used yielded similar results in composition as well as relative abundance at phyla (Proteobacteria, Firmicutes, and Bacteroidota) and genus (Klebsiella, Mycoplasma, and Aeromonas) levels. A significant difference was found in alpha diversity (Chao1) between LVL and endometrial biopsy, suggesting that LVL is superior at detecting the low-abundant (rare) taxa. These new data could pave the way for innovative treatment methods for endometrial disease and subfertility in mares. This, in turn, could lead to more judicious antimicrobial use in the equine breeding industry.

Comparing the pharmacokinetics of a fourth generation cephalosporin in three different age groups of New Forest ponies.

REASONS FOR PERFORMING STUDY: To compare the pharmacokinetics of the fourth generation cephalosporin, cefquinome, in neonatal foals, 6-week-old foals and mature New Forest ponies in order to recommend appropriate dosage regimens for use of this drug. METHODS: Cefquinome was administered i.v. at 1 mg/kg bwt twice a day (q. 12 h), 1 mg/kg bwt 3 times a day (q. 8 h) or 4.5 mg/kg bwt q. 12 h to each age group (n = 6). Plasma cefquinome concentrations were analysed using high-performance liquid chromatography combined with electrospray tandem mass spectrometry. RESULTS: Both foal age groups had comparable pharmacokinetic data except for the volume of distribution at a steady-state (Vss), total body clearance (ClB) and mean residence time (MRT). Both ClB and MRT decreased as the age of the foals increased. Values of area under the curve increased, in a dose dependent manner, with significant increases for all age groups following administration of 4.5 mg/kg bwt q. 12 h. Total body clearance did not have comparable dose dependency. CONCLUSIONS: Cefquinome can be given at a dose of 1 mg/kg bwt q. 12 h for the treatment of infections caused by susceptible pathogens with MIC < 0.125 microg/ml. A higher dose of 4.5 mg/kg bwt q. 12 h is recommended for the treatment of bacterial pathogens with minimal inhibitory concentration (MIC) 0.125-0.5 microg/ml POTENTIAL RELEVANCE: Commonly used dosing regimens should be critically evaluated in neonatal foals due to the higher volume of distribution of less lipophilic drugs in this age group.

Contribution of age and gender to outcome of blunt splenic injury in adults: multicenter study of the eastern association for the surgery of trauma.

BACKGROUND: The purpose of this study was to examine the contribution of age and gender to outcome after treatment of blunt splenic injury in adults. METHODS: Through the Multi-Institutional Trials Committee of the Eastern Association for the Surgery of Trauma (EAST), 1488 adult patients from 27 trauma centers who suffered blunt splenic injury in 1997 were examined retrospectively. RESULTS: Fifteen percent of patients were 55 years of age or older. A similar proportion of patients > or = 55 went directly to the operating room compared with patients < 55 (41% vs. 38%) but the mortality for patients > or = 55 was significantly greater than patients < 55 (43% vs. 23%). Patients > or = 55 failed nonoperative management (NOM) more frequently than patients < 55 (19% vs. 10%) and had increased mortality for both successful NOM (8% vs. 4%, p < 0.05) and failed NOM (29% vs. 12%, p = 0.054). There were no differences in immediate operative treatment, successful NOM, and failed NOM between men and women. However, women > or = 55 failed NOM more frequently than women < 55 (20% vs. 7%) and this was associated with increased mortality (36% vs. 5%) (both p < 0.05). CONCLUSION: Patients > or = 55 had a greater mortality for all forms of treatment of their blunt splenic injury and failed NOM more frequently than patients < 55. Women > or = 55 had significantly greater mortality and failure of NOM than women < 55.

13C and 15N distributions in three spodic dystric cambisols under beech and spruce.

The study of natural isotopic abundance signatures is useful to gain further insights in the processes resulting in depthwise changes in the composition of soil organic matter (SOM). Objectives were to describe the delta 13C and delta 15N abundances of SOM with depth in soils from a 153-year old beech (B1), a 119-year old spruce (F1) and a 61-year old spruce (F2) stand at Solling, north-west Germany, and to study, how podzolisation affects the isotopic abundances of 13C and 15N in the SOM. The degree of podzolisation decreased in the order F1 > B1 > F2. At the surface of the humus layer of all three sites, delta 13C values are approximately 1 to 4/1000 higher than in the leaves and needles, probably mainly due to the discrimination of 13C by microbial decomposition. 13C abundances in the organic layers of B1 and F2 increased only slightly from -27.6/1000 PDB (B1, L) to -27.2/1000 PDB (B1, Oh) and from -26.3/1000 PDB (F2, L) to -25.9/1000 PDB (F2, Oh), suggesting that biotic activity resulted in mixing of organic matter. At F1, however, 13C abundance increased from -27.5/1000 PDB (L) to -26.0/1000 PDB (Oh) which reflects the lack of mixing by animals. In the upper 2-4 cm of the mineral soil, i.e., in the eluvial horizons Aeh, 13C values showed a minimum at the spruce sites which was presumably related to a translocation of 13C enriched fulvic acids. Depthwise changes in delta 15N values were not related to podzolisation processes. At all three sites, a 13N enrichment with depth occurred in the mineral soil which is the result of the discrimination of 15N by microbial decomposition.

Use of 13C and 15N mass spectrometry to study the decomposition of Calamagrostis epigeios in soil column experiments with and without ash additions.

The dynamics of C and N in terrestrial ecosystems are not completely understood and the use of stable isotopes may be useful to gain further insight in the pathways of CO2 emissions and leaching of dissolved organic carbon (DOC) and nitrogen (DON) during decomposition of litter. Objectives were (i) to study the decomposition dynamics of Calamagrostis epigeios, a common grass species in forests, using 13C-depleted and 15N-enriched plants and (ii) to quantify the effect wood ash addition on the decomposition and leaching of DOC and DON. Decomposition was studied for 128 days under aerobic conditions at 8 degrees C and moisture close to field capacity in a spodic dystric Cambisol with mor-moder layer. Variants included control plots and additions of (i) Calamagrostis litter and (ii) Calamagrostis litter plus 4 kg ash m-2. (i) Decomposition of Calamagrostis resulted in a CO2 production of 76.2 g CO2-C m-2 (10% of added C) after 128 days and cumulative DOC production was 14.0 g C m-2 out of which 0.9 g C m-2 was Calamagrostis-derived (0.1% of added C). The specific CO2 formation and specific DOC production from Calamagrostis were 6 times higher (CO2) and 4 times smaller (DOC) than those from the organic layer. The amount of Calamagrostis-derived total N (NH4+, NO3-, DON) leached was 0.7 g N m-2 (4.8% of added N). Cumulative DON production was 0.8 g N m-2 which was slightly higher than for the control. During soil passage, much of the DOC and DON was removed due to sorption or decomposition. DOC and DON releases from the mineral soil (17 cm depth) were 6.3 g C m-2 and 0.5 g N m-2. (ii) Addition of ash resulted in a complete fixing of CO2 for 40 days due to carbonatisation. Afterwards, the CO2 production rates were similar to the variant without ash addition. Production of DOC (98.6 g C m-2) and DON (2.5 g N m-2) was marked, mainly owing to humus decay. However, Calamagrostis-derived DOC and Calamagrostis-derived total N were only 3.9 g C m-2 (0.5% of added C) and 0.5 g N m-2 (3.4% of added N). The specific DOC production rate from the organic layer was 6 times higher than that from Calamagrostis. The results suggest that with increasing humification from fresh plant residues to more decomposed material (OF and OH layers) the production ratio of DOC/CO2-C increases. Addition of alkaline substances to the forest floor can lead to a manifold increase in DOC production.

Blunt splenic injury in adults: Multi-institutional Study of the Eastern Association for the Surgery of Trauma.

BACKGROUND: Nonoperative management of blunt injury to the spleen in adults has been applied with increasing frequency. However, the criteria for nonoperative management are controversial. The purpose of this multi-institutional study was to determine which factors predict successful observation of blunt splenic injury in adults. METHODS: A total of 1,488 adults (>15 years of age) with blunt splenic injury from 27 trauma centers in 1997 were studied through the Multi-institutional Trials Committee of the Eastern Association for the Surgery of Trauma. Statistical analysis was performed with analysis of variance and extended chi2 test. Data are expressed as mean +/- SD; a value of p < 0.05 was considered significant. RESULTS: A total of 38.5 % of patients went directly to the operating room (group I); 61.5% of patients were admitted with planned nonoperative management. Of the patients admitted with planned observation, 10.8% failed and required laparotomy; 82.1% of patients with an Injury Severity Score (ISS) < 15 and 46.6% of patients with ISS > 15 were successfully observed. Frequency of immediate operation correlated with American Association for the Surgery of Trauma (AAST) grades of splenic injury: I (23.9%), II (22.4%), III (38.1%), IV (73.7%), and V (94.9%) (p < 0.05). Of patients initially managed nonoperatively, the failure rate increased significantly by AAST grade of splenic injury: I (4.8%), II (9.5%), III (19.6%), IV (33.3%), and V (75.0%) (p < 0.05). A total of 60.9% of the patients failed nonoperative management within 24 hours of admission; 8% failed 9 days or later after injury. Laparotomy was ultimately performed in 19.9% of patients with small hemoperitoneum, 49.4% of patients with moderate hemoperitoneum, and 72.6% of patients with large hemoperitoneum. CONCLUSION: In this multicenter study, 38.5% of adults with blunt splenic injury went directly to laparotomy. Ultimately, 54.8% of patients were successfully managed nonoperatively; the failure rate of planned observation was 10.8%, with 60.9% of failures occurring in the first 24 hours. Successful nonoperative management was associated with higher blood pressure and hematocrit, and less severe injury based on ISS, Glasgow Coma Scale, grade of splenic injury, and quantity of hemoperitoneum.

Helmet and shoulder pad removal from a player with suspected cervical spine injury. A cadaveric model.

STUDY DESIGN: Video fluoroscopy was used to evaluate the motion in an unstable spine during helmet and shoulder pad removal. OBJECTIVE: To observe the amount of motion that occurs during the removal of helmet and shoulder pads in an injured spine. SUMMARY OF BACKGROUND DATA: Removal of shoulder pads and helmet from a football player with suspected cervical spine injury can be particularly hazardous. How much flexion occurs at the unstable level during removal of equipment is unknown. METHODS: Six fresh cadavers were used in the study. In three, an unstable C1-C2 segment was created by transoral osteotomy of the base of C2. In the remaining three, instability was created at C5-C6 by a posterior release. Under fluoroscopic recording, the helmets were removed by first removing the chin strap, face mask, and ear pieces. With the neck stabilized, the helmet was carefully removed. The shoulder pads were carefully removed, with the head stabilized. Angulation, distraction, and space available for the cord were measured at C1-C2. Translation, angulation, distraction, and change in disc height were measured in the specimens with unstable C5-C6. RESULTS: In cadavers with C1-C2 instability, the mean change in angulation was 5.47 degrees, and space available for the cord was 3.91 mm. Shoulder pads were removed while the head was stabilized. The mean change in angulation at C1-C2 was less during removal of shoulder pads than during helmet removal at 2.9 degrees. Space available for the cord was 2.64 mm. Distraction was also greater during helmet removal (2.98 mm) than during shoulder pad removal (1.76 mm). In the unstable spine, the change in displacement in translation was greater during shoulder pad removal (3.87 mm), than during helmet removal (0.41 mm). Disc height change was similar. Distraction of the spinous processes was greater during helmet removal (3.68 mm) than during shoulder pad removal (1.37 mm). Angulation was similar in both maneuvers. CONCLUSIONS: Helmet and shoulder pad removal in the unstable cervical spine is a complex maneuver. In the unstable C1-C2 segment, helmet removal causes more angulation in flexion, more distraction, and more narrowing of the space available for the cord. In the lower cervical spine (C5-C6), helmet removal causes flexion of 9.32 degrees, and during shoulder pad removal the neck extends 8.95 degrees, a total of approximately 18 degrees. Disc height changes from 1.24 mm of distraction to 1.06 mm of compression during helmet removal and shoulder pad removal for a total 2.3-mm change. Translation, which correlates with the change in the space available for the cord, is greater at C5-C6 during shoulder pad removal. Because most of the cadavers had C5 anteriorly displaced on C6 to begin with, the extension force during shoulder pad removal caused a 3.87-mm change in reduction of C5 on C6. Because of the motion observed in the unstable spine, helmet and shoulder pad removal should be performed in a carefully monitored setting. They should be removed together by at least three, preferably four, trained people.

Application of thrombin based fibrin glue and non-thrombin based batroxobin glue on intact human blood vessels: evidence for transmural thrombin activity.

An alternative method of uniting small diameter vessels to obtain tissue union while limiting the thrombogenic effect of suture placement at a vessel anastomosis involves the use of a thrombin based fibrin glue as a surgical sealant. This investigation addresses whether the in vitro application of a thrombin based glue (TG), or batroxobin glue (BG), a non-thrombin based glue made with the snake venom enzyme batroxobin, alters intravascular platelet deposition (PD) or cleaves blood fibrinogen, as measured by fibrinopeptide A (FPA) production, when the respective glue is applied to the external surface of an intact human placental artery or an artery with an anastomosis. When TG was applied to the adventitial surface of an intact vessel or an anastomosis (n = 7) of control and experimental vessels, there was a significant increase in intraluminal platelet deposition, an effect not realized with BG (n = 12, intact vessel TG p = 0.01, BG p = 0.66, anastomosis TG p <0.01, BG p <0.01). Both TG and BG significantly increased FPA levels when human whole blood was perfused through both intact vessels or vessels containing an anastomosis when compared to control vessels (intact vessel TG and BG p <0.01, anastomosis TG and BG p <0.01). Labelled thrombin studies document the rapid passage of thrombin through an intact vessel wall or vessels with an anastomosis when TG was applied to the adventitial surface of the vessel. The data suggest that TG and BG are drug delivery systems for their respective enzymes that either pass through or transfer a message across not only a surgically created anastomosis, but also an intact vessel wall.

The effect of airway maneuvers on the unstable C1-C2 segment. A cadaver study.

STUDY DESIGN: This is a cadaver study in which video fluoroscopy is used to measure motion of the unstable spine at C1-C2 during intubation maneuvers. OBJECTIVES: To quantify the amount of motion that occurs at an unstable C1-C2 spinal segment during the use of various intubation techniques using a cadaver model. SUMMARY OF BACKGROUND DATA: In previous work by the authors, a methodology and measurements for the unstable C5-C6 segment in a cadaver model were developed. These studies showed that the most motion was created by a chin lift and jaw thrust and that oral techniques created more motion than nasal intubation. The potential motion that occurs during intubation with instability at C1-C2 is yet unstudied. Therefore, a study to determine the effects of intubation on the spine with an unstable C1-C2 segment was designed. METHODS: Six human cadavers were used for the study. Measurements before and after transoral osteotomy of the odontoid were performed using video fluoroscopy. Pre-intubation maneuvers and oral and nasal intubation were studied. RESULTS: Oral intubation and nasal intubation caused similar diminution of space available for the cord. Chin lift and jaw thrust caused a larger diminution of space available for the cord than either nasal or oral intubation techniques. CONCLUSIONS: Although nasal intubation is the accepted procedure for intubation of the unstable spine, nasal and oral intubation seemed to have the same ability to narrow the space available for the cord in the model in this study. Great care should be taken while performing the chin lift/jaw thrust maneuvers in preparation for intubation, because these pre-intubation techniques caused the most motion and hence narrowed the space available for the cord in the unstable cervical spine.

Thrombin activity on dissected and anastomosed human arteries.

The mechanism of anastomotic thrombosis in microvascular surgery remains poorly understood. We hypothesized that thrombin activity at anastomoses plays a major role in this process. To study this, a surgically relevant human artery anastomosis model was used to (i) measure surface thrombin activity on anastomoses and on intact vessel, (ii) determine the inhibitability of surface thrombin by heparin and recombinant hirudin (r-hirudin), and (iii) determine the anastomotic and intact vessel binding capacity for additional thrombin. Human placental artery segments were placed in chambers in which 0.2 cm2 of luminal surface was exposed to citrated platelet-poor plasma for 10 min at 37 degrees C. The fibrinopeptide A (FPA) concentration (indicating the action of thrombin on fibrinogen) in the supernatant was then measured using an ELISA assay. Intact vessels and anastomoses expressed equivalent thrombin activity that could not be inhibited by heparin at a concentration (0.3 U/ml) that is sufficient to prolong the activated partial thromboplastin time two-fold. Conversely, the concentration of heparin routinely used in intraoperative vessel irrigation solutions (50 U/ml) was able to completely block thrombin activity at both sites. r-Hirudin (0.3 heparin equivalent anti-IIa U/ml) was able to inhibit nearly all of the thrombin activity on each site. Each site was able to bind and express the activity of additional thrombin, indicating the potential for increased vessel thrombogenicity after local clot has formed and has been removed. These data indicate the presence of thrombin on dissected human vessels and its presence in equal amounts on intact and anastomosed vessels when measurement is made before blood flow resumes. Furthermore, vessel-associated thrombin is resistant to a standard systemic concentration of heparin but is susceptible to the much higher heparin concentration that can be delivered locally by the surgeon during vessel irrigation.

Tissue factor and its inhibition at the human microvascular anastomosis.

The surgically created vascular anastomosis is a thrombogenic zone of uncertain etiology. This study was designed to investigate the importance of tissue factor as a cause of human microvascular thrombogenicity. The ability of tissue factor pathway inhibitor (TFPI) to block the effect of tissue factor was also tested in this whole-vessel model system. Tissue factor activity in the presence of absence of TFPI was assayed on the luminal surface of dissected human placental arteries, on the advential surface, and also at the site of a microvascular anastomosis. Vessel wall thrombin activity was measured in the presence and absence of TFPI. Platelet deposition onto a vessel surface using a perfusion system was measured with and without TFPI. Tissue factor activity was greater on the adventitia (4.6 +/- 2.8 x 10(-4) units factor Xa generated/min) than on the endothelium (1.8 +/- 1.6 x 10(-4), P < 0.03) or at a surgically created anastomosis (2.1 +/- 1.2 x 10(-4), P < 0.04). TFPI reduced Xa generation to undetectable levels in 21 of 23 endothelial, adventitial, and anastomotic segments (P < 0.002). TFPI significantly reduced vessel wall thrombin activity in comparison to control anastomoses (control, 3.2 +/- 1.7 ng fibrinopeptide A (FPA)/(ml x min); TFPI, 1.4 +/- 1.2 ng FPA/(ml x min); P < 0.0001). TFPI reduced the platelet deposition on vessel segments with intact endothelium (no TFPI, 0.88 +/- 0.69 x 10(6) platelets/cm2; TFPI, 0.49 +/- 0.29 x 10(6) platelets/cm2; P < 0.06) and on vessel segments with anastomoses (no TFPI, 1.3 +/- 0.70 x 10(6) platelets/cm2; TFPI, 0.76 +/- 0.35 x 10(6) platelets/cm2; P < 0.02). This study demonstrates the importance of tissue factor as a thrombogenic element in a human whole-vessel model system. TFPI is effective in reducing this thrombogenicity.

Positron emission tomography within a magnetic field using photomultiplier tubes and lightguides.

The spatial resolution of positron emission tomography (PET) improves when positron annihilation takes place in a strong magnetic field. In a magnetic field, the Lorentz force restricts positron range perpendicular to the field. Since positron annihilation occurs closer to its point of origin, the positron annihilation point spread function decreases. This was verified experimentally by measuring the spread function of positron annihilation from a 500 mm 68Ge bead imbedded in tissue-equivalent wax. At 5 T the spread function full width at half maximum (FWHM) and the full width at tenth maximum (FWTM) decrease by a factor of 1.42 and 2.09, respectively. Two NaI(Tl) scintillation crystals that interface to a pair of photomultiplier tubes (PMTS) through long lightguides detect positron annihilation at zero field and 5.0 T. Photomultiplier tubes, inoperable in strong magnetic fields, are functional if lightguides bring the photons produced by scintillators within the field to a minimal magnetic field. These tests also demonstrate techniques necessary for combining magnetic resonance imaging (MRI) and PET into one scanner.

Use of a magnetic field to increase the spatial resolution of positron emission tomography.

Detector geometry, spatial sampling, and more fundamentally, positron range and noncollinearity of annihilation photon emission define Positron Emission Tomography (PET) spatial resolution. In this paper, a strong magnetic field is used to constrain positron travel transverse to the field. Measurement of the spread function from a 500 microns diameter 68Ga impregnated resin bead shows a squeezing of the full width at half maximum (FWHM) by a factor of 1.0, 1.22, 1.42, and 2.05, at 0, 4.0, 5.0, and 9.4 Tesla, respectively. The full width at tenth maximum (FWTM) decreases by a factor of 1.0, 1.73, 2.09, and 3.20, at 0, 4.0, 5.0, and 9.0 Tesla, respectively. Acquiring a PET image in a magnetic field should significantly reduce resolution loss due to positron range.

Insulin therapy increases low plasma growth hormone binding protein in children with new-onset type 1 diabetes.

This study was undertaken (1) to evaluate growth hormone binding protein (GHBP) levels in newly diagnosed patients with Type 1 diabetes before and after insulin therapy and (2) to determine the relationship of GHBP to glycaemic control, C-peptide level and blood pH. GHBP, expressed as a percentage of (125I)GH bound, was determined in 33 patients with Type 1 diabetes (M/F = 19/14, 12.3 +/- 0.4 years) before (day 0), after 5 days (day 5) and after 3 months (month 3) of insulin therapy. At day 0, GHBP was lower in Type 1 diabetes compared with 38 matched healthy control subjects (3.9 +/- 0.4 vs 8.2 +/- 0.4%, p < 0.001). There was no significant improvement in GHBP at day 5 (4.4 +/- 0.3%). At month 3, GHBP increased to (6.0 +/- 0.4%, p < 0.001 vs day 0), but was still lower than controls, p < 0.001. At day 0 GHBP correlated with BMI (r = 0.50, p = 0.001), blood glucose (r = -0.43 p = 0.006) and pH (r = 0.48, p = 0.004), but not HbA1. GHBP at month 3 correlated with day 0 C-peptide (r = 0.41, p = 0.02). Thus, (1) circulating GHBP is low in newly diagnosed patients with Type 1 diabetes, and increases after 3 months of insulin therapy but does not normalize and (2) the severity of biochemical derangement and residual beta-cell function at diagnosis may determine GHBP status and its recovery. We conclude that insulin is an important modulator of GH binding protein in newly diagnosed children with Type 1 diabetes.

Hepatic insulin action in adolescents with insulin-dependent diabetes mellitus: relationship with long-term glycemic control.

The relationship between hepatic insulin action and long-term glycemic control was assessed in 20 adolescents with insulin-dependent diabetes mellitus (IDDM) and five healthy matched controls using a two-step (0.8 and 1.6 mU/kg/min) hyperinsulinemic-euglycemic clamp and [6,6-2H2]glucose. The night before the study, diabetic patients received variable-rate intravenous insulin in an attempt to normalize fasting plasma glucose concentrations. In the postabsorptive state, hepatic glucose production (HGP) was similar in IDDM patients and controls (593 +/- 40 v 518 +/- 27 mumol/m2/min); however, plasma glucose and free insulin concentrations were higher in IDDM patients than in controls (6.5 +/- 0.4 v 5.4 +/- 0.1 mmol/L [P = .01], and 207 +/- 21 v 104 +/- 10 pmol/L [P < .001], respectively). There was a positive correlation (r = .62 P = .002) between basal HGP and glycohemoglobin level (HbA1). Separation of the patients at the median HbA1 (group no. 1 < and group no. 2. > 11.4% HbA1) revealed two distinct patient populations with regard to hepatic and peripheral insulin action and plasma free fatty acid (FFA) suppression. During hyperinsulinemia, the percent suppression of HGP was lower in group no. 2 compared with group no. 1 (65.7% +/- 9.8% v 94.4% +/- 3.8%; P = .018). Rates of glucose disposal were lower in group no. 2 compared with controls and with group no. 1. Postabsorptive FFA levels were similar between group no. 2 and group no. 1 (0.45 +/- 0.03 and 0.43 +/- 0.04 mmol/L) despite higher free-insulin concentrations in group no. 2 (260 +/- 30 v 171 +/- 25 pmol/L; P = .04).(ABSTRACT TRUNCATED AT 250 WORDS)

Synergy between tetrandrine and FK506 in prevention of diabetes in BB rats.

Delayed administration of tetrandrine, a novel broad-spectrum anti-inflammatory agent, to BB rats at a dosage schedule of 20 mg kg-1 day-1 from 79 days of age reduced the cumulative incidence of diabetes from 73.1 to 41.7% (p < 0.01). Brief treatment with the potent immunosuppressive agent FK506 at a dosage schedule of 0.5 mg kg-1 day-1 from 79 days of age for 5 days had no significant effect on the cumulative incidence of diabetes (66.7%, p > 0.1). However, the combination of tetrandrine and FK506 in the afore-mentioned dosage schedules reduced the incidence of diabetes to only 3.6% (p < 0.001). These results suggest that the strong synergy between tetrandrine and FK506 may offer a safe and effective therapeutic strategy for the treatment of patients with recent onset or imminent IDDM.