A refined method for assessing 99mTc-MDP whole body retention in prostate cancer patients

Whole body retention (WBR) of ^99mTc labeled methylene diphosphonate (MDP) has been shown to significantly differentiate various clinical stages of prostate cancer. Whole body measurements, when performed at 5 min and 24 h after i.v. administration of ^99mTc-MDP, allows for the calculations of percentage whole body retention (% WBR) after one day. The latter can be expressed relative to either the anterior or posterior projection, or in combination as the geometric mean value.In an attempt to better describe the clinical course of prostate cancer patients with bone metastases we have refined the % WBR calculations to include a normalization factor. The latter consists of the mean 24-h value of WBR's as obtained from 10 prostate cancer patients without bony métastases as determined by bone scintigram. These values were determined for each projection to be: anterior = 26.5 ± 4.7%, posterior = 37.5 ± 7.4%, and geometric mean = 31.5 ± 5.7%. The % WBR is then divided by the normalization factor of choice and expressed as (% WBR)^N.These data are used to better express ^99mTc MDP 24-h whole body retentions when following the clinical course of patients with metastatic carcinoma of the prostate. Caution should be exercised when interpreting these data when metabolic bone pathology is present. A false negative (% WBR)^N value will result if an infiltration of the ^99mTc-MDP occurs during administration.     

Molar proportions and relative rates of synthesis of histones in rat testis

The molar proportions and relative rates of synthesis of histones in normal and hypophysectomized rat testis seminiferous epithelial cells were determined. After hypophysectomy the molar proportions of histones H1, H2B and (H2A + protein A24) in seminiferous epithelial cells of rat testis increased while their corresponding variants TH1-x, TH2B-x and X₂ decreased, but the molar proportions of major-class histones (i.e., sum of subfractions) remained relatively constant and similar to the proportions in somatic cells. The apparent molar proportions of the labeled histones, determined immediately after 2-h periods of [³H]leucine incorporation, were much higher relative to H4 than the proportions of total histones determined by dye binding. The values, however, approached the molar proportions of total histones when rats were killed 11 days after the [³H]leucine injection. Two-dimensional gel electrophoresis confirmed that the high initial molar proportions relative to H4 by [³H]leucine incorporation were not due to the possible contamination by highly-labeled non-histone proteins. The specific activity of histone H4 relative to the specific activity of DNA, determined immediately after 3-h periods of [³H]leucine and [¹⁴C]thymidine incorporations was similar to the value when rats were killed 13 days after the injections. It is proposed that histones of seminiferous epithelial cells are synthesized disproportionally relative to H4 and in excess of the quantities required for polynucleosome assembly. The excess histones are subsequently displaced or degraded slowly.     

High Dietary Sodium Intake Assessed by Estimated 24-h Urinary Sodium Excretion Is Associated with NAFLD and Hepatic Fibrosis

Background

Although high sodium intake is associated with obesity and hypertension, few studies have investigated the relationship between sodium intake and non-alcoholic fatty liver disease (NAFLD). We evaluated the association between sodium intake assessed by estimated 24-h urinary sodium excretion and NAFLD in healthy Koreans.

Methods

We analyzed data from 27,433 participants in the Korea National Health and Nutrition Examination Surveys (2008–2010). The total amount of sodium excretion in 24-h urine was estimated using Tanaka’s equations from spot urine specimens. Subjects were defined as having NAFLD when they had high scores in previously validated NAFLD prediction models such as the hepatic steatosis index (HSI) and fatty liver index (FLI). BARD scores and FIB-4 were used to define advanced fibrosis in subjects with NAFLD.

Results

The participants were classified into three groups according to estimated 24-h urinary excretion tertiles. The prevalence of NAFLD as assessed by both FLI and HSI was significantly higher in the highest estimated 24-h urinary sodium excretion tertile group. Even after adjustment for confounding factors including body fat and hypertension, the association between higher estimated 24-h urinary sodium excretion and NAFLD remained significant (Odds ratios (OR) 1.39, 95% confidence interval (CI) 1.26–1.55, in HSI; OR 1.75, CI 1.39–2.20, in FLI, both P < 0.001). Further, subjects with hepatic fibrosis as assessed by BARD score and FIB-4 in NAFLD patients had higher estimated 24-h urinary sodium values.

Conclusions

High sodium intake was independently associated with an increased risk of NAFLD and advanced liver fibrosis.     

Endogenous 24-hour cycle of core temperature and oxygen consumption in week-old Zucker rat pups

Summary Experiments were designed to test whether or not the 24-h core temperature fluctuations in week-old rat pups are of endogenous origin. Lean (Fa/-) Zucker rat pups born on the same day to mothers maintained in two different colonies with light/dark cycles 12 h out of phase with each other were mother-reared through the first 3–4 days of life and then artificially reared simultaneously in constant dim light. Continuous, automatic measurement of core temperature and oxygen consumption during artificial rearing showed clear 24-h rhythms in 5- to 8-day-old pups. Each rhythm reached a daily minimum at a time corresponding to the beginning of the light period in the colony of origin. The amplitude of these rhythms did not diminish during artificial rearing, nor did the phase difference between the rhythms of pups originating in the two colonies systematically change. The persistent 12-h phase differences between these two groups of pups prove that the observed rhythms are not caused by exogenous stimuli. We conclude that the rat pup possesses an endogenous time-keeping mechanism that permits the expression of overt rhythmicity at the age of 1 week.Abbreviations Tc core temperature - LD-pups born to a mother entrained to a 7:00 to 19:00 light cycle, then artificially reared in constant dim light - DL-pups born to a mother entrained to a 19:00 to 7:00 light cycle, then artificially reared in constant dim light - SCN suprachiasmatic nucleus     

The effect of age on beta-adrenergic receptors and adenylate cyclase activity in rat erythrocytes.

Beta-adrenergic receptors and catecholamine-sensitive adenylate cyclase activity were studied in erythrocytes obtained from rats 6 weeks, 6 months, and 15 months of age. Intact erythrocytes from 6 week old rats contained significantly more beta receptors (411 ± 31 sites/cell) than 6 month (328 ± 21) or 15 month old rats (335 ± 16), as determined by binding of [¹²⁵I] iodohydroxybenzylpindolol. Erythrocytes from 6 week old rats also contained significantly greater isoproterenol-sensitive adenylate cyclase activity (95.0 ± 9.4pmoles/10⁹ cells) than erythrocytes from 6 month (27.9 ± 3.3) or 15 month old rats (23.7 ± 3.6). The erythrocyte population of 6 week old rats was bigger (mean corpuscular volume = 62 ± 2μ^3/cell) than the older rat erythrocytes (47 ± 1μ³ and 48 ± 1μ³). When the data were expressed relative to a unit of cell volume, there was no difference in the density of beta receptors among all three populations but a progressive and significant fall in hormone-sensitive adenylate cyclase activity. In the rat erythrocyte, the age-related loss of adenylate cyclase activity is not accompanied by changes in β-receptor density.     

In vitro growth response of Prunus cerasifera shoots as influenced by different light-dark cycles and sucrose concentrations

Trials were carried out to test if the higher growth response shown by shoot clusters of Mr. S. 2/5, a clonal selection of Prunus cerasifera, submitted to short and frequent light-dark regimes could be related to the amount of sucrose added to growth medium.The reduction of sucrose from 30 gl^-1 (control) to 22.5 gl^-1, 15 gl^-1 and 7.5 gl^-1 caused a progressive and remarkable inhibition of shoot tip growth. With 15 gl^-1 the value of some growth parameters was reduced by more than half. Under 16-h daylength, the best sucrose concentration was 30 gl^-1, while with 4-h light-2-h dark no statistical differences appeared between 30 gl^-1 and 22.5 gl^-1 sucrose. Compared to 16-h light-8-h dark, the 4-h light-2-h dark cycle at the three highest sucrose concentrations gave rise to higher values of fresh and dry weight as well as increasing the number of axillary shoots produced.The increment in growth response induced by the shorter light-dark regime decreased with diminishing growth capacity in the cultures when sucrose concentration was lowered, but it was still appreciable even with 7.5 gl^-1. Since the 4-h light-2-h dark cycle induced a favourable effect in culture growth with all sucrose concentrations, we conclude that the greater growth response observed with this light regime was not triggered by carbohydrate availability but by some other unknown factors.     

Efficacy and safety of Abelmoschus manihot for IgA nephropathy: A multicenter randomized clinical trial

Rationale and ObjectiveIgA nephropathy (IgAN) is an important cause for end-stage renal disease worldwide. The treatment for IgAN remains challenging, and few randomized and controlled clinical trials have been conducted to evaluate new therapies. The present study assesses the efficacy and safety of Abelmoschus manihot (AM) in IgAN patients.Study DesignRandomized, non-inferiority, double-blind, double-dummy multicenter trial.Setting and ParticipantsThis trial was designed to recruit 1,600 biopsy-proven IgAN patients (proteinuria between 0.5-3.0 g/d and estimated glomerular filtration rate [eGFR] of ≥ 45 ml/min/1.73 m²) across China.InterventionsThe participants were randomized at 1:1 to AM (2.5 g for three times per day) or losartan potassium (100 mg per day) for 48 weeks.OutcomesThe primary outcome was the change in 24-hour proteinuria from baseline to week 48. The secondary outcomes were the change in eGFR from baseline to week 48, and the incidents of endpoint events (proteinuria ≥ 3.5 g/24 h, doubling of serum creatinine, or receiving renal replacement treatment).ResultsAmong 1,470 randomized patients (mean age, 37.4 [SD, 10.6] years old; 777 [52.9%] were female; mean eGFR, 95.0 [SD, 24.3] mL/min/1.73 m²; mean 24-hour proteinuria, 1.2 [SD, 0.7] g/d), the mean decline in 24-h proteinuria at week 48 was 230 mg and 253 mg in the AM and losartan potassium groups, respectively (P = 0.676). The mean difference in the change in 24-h proteinuria between these two groups was -23.32 mg (95% confident interval: -123.2 to 76.6, p = 0.647). The mean decline in eGFR was 0.41 ml/min/1.73 m² and 0.76 ml/min/1.73 m² in the AM and losartan potassium groups, respectively (p = 0.661). The mean difference in the change in eGFR between these two groups was -0.43 ml/min/1.73 m² (95% confident interval: -1.99 to 1.13, p = 0.589). The incidence of endpoint events was 8.6% in the AM group and 8.2% in the losartan group (p = 0.851).LimitationsThe results of the trial may not be generalized to IgAN patients with a proteinuria of > 3.0 g/d and an eGFR of < 45 ml/min/1.73 m². The long-term benefits of AM in reducing the risk of progressive renal dysfunction remains unclear, based on this 48-week observation.ConclusionAM can be recommended as a promising treatment for IgAN patients.     

p NMR Studies of Rat Liver Cold Preservation with Histidine-Buffered Lactobionate Solution

The efficiency of a preservation medium, histidine-buffered lactobionate solution (HBLS), was determined by measuring post-ischemic recoveries of ATP and intracellular pH under Krebs-Henseleit buffer (KHB) perfusion. We used NMR spectroscopy to study the effect of 24-h cold ischemia, followed by 4°C then 37°C reperfusion on the isolated rat liver. Three media were compared: University of Wisconsin solution (UW-lactobionate); Bretschneider's solution (HTK); HBLS and HBLS supplemented with 2 mM Gly and 2 mM Cys (HBLSg2) or with 10 mM Gly and 2 mM Cys (HBLSg10). All values were compared to control values measured during pre-ischemic cold perfusion with KHB (ATP = 8.60 ± 0.6 μmol/g of dry weigh and pH_in = 7.41 ± 0.05). The main result from ³¹p NMR data concerned ATP recovery during cold reperfusion, which was significantly higher in the HBLS group (112 ± 10%) as compared to the UW and HTK groups (around 66%). The presence of glycine decreased ATP recovery (88 ± 8% in HBLSg2, 79 ± 15% in HBLSg10). Higher values of recovered pH_in were observed in livers stored in histidine buffered solutions (around 7.30) as compared to UW (around 7.20); histidine was by ¹³C NMR proved to accumulate in the liver cells, thus ensuring a good buffering capacity. The thermal transition induced a decrease in both ATP level and pH_in in all groups. This might be the result of a stimulation of the carbohydrate metabolism (as demonstrated by ¹³C NMR) especially when glycine was present in the storage solution.     

Variability in nitrogen fixation of common bean (Phaseolus vulgaris L.) intercropped with maize

Thirty one selected bean lines were evaluated in the field for ability to support N₂ fixation when intercropped with maize which received 0, 30 and 60 kg N ha^–1 as ammonium sulphate. The amount of fixed N₂ was estimated using the natural variation of ¹⁵N and wheat as the standard non-fixing crop. Nitrogen as low as 15 kg N ha^–1 at sowing suppressed nodule weight and activity (acetylene reduction activity) but not nodule number, suggesting that the main effect of mineral N was on nodule development and function. ¹⁵N data revealed a high potential of the bean genotypes to fix N₂, with the most promising ones averaging between 50–60% of seed N coming from fixation. Bean lines CNF-480, Puebla-152, Mexico-309, Negro Argel, CNF-178, Venezuela-350 and WBR22-3, WBR22-50 and WBR22-55 were ranked as good fixers.     

Strength training for 1h in humans: effect on the motor performance of normal upper extremities

It has been found that one session of intense muscle strength training decreases muscle strength temporarily and causes neuromuscular fatigue in the trained muscles, but little attention has been given to the effects of neuromuscular fatigue on the other components of motor performance. The purpose of this study was to examine in normal healthy volunteers the effects of a 1-h strength training session on the motor performance of the upper extremity, including reaction time, speed of movement, tapping speed and coordination. Group of 30 healthy female volunteers, aged 29-47 years, were randomly divided into sub-groups, (A and B, n = 15 per group). Both groups first completed a set of motor performance tests on 3 consecutive days. On the 4th day, group A carried out a 1-h muscle strength training session of the upper extremities. Isometric muscle strengths and electromyogram (EMG) data were recorded before the training session. Immediately after the training session the same recordings were repeated, and additional motor performance tests were also performed. Group B carried out only the motor performance tests. The groups exchanged programmes the following week. The 1-h strength training session decreased the isometric muscle strength of wrist flexion by 18% (P < 0.001) and extension by 18% (P < 0.001) in group A, while in group B flexion strength decreased by 19% (P < 0.001) and extension strength by 17% (P < 0.001). All the measured EMG activations also decreased in both groups. There were no statistically significant differences in the results of the motor performance tests between the mean values of the three baseline measurements and the values recorded after the training session. The result was surprising, but straightforward; neuromuscular fatigue induced by a 1-h strength training session of the upper extremities had no effect on the motor performance functions of the hand, as indicated by reaction times, speed of movement, tapping speed and coordination, in these normal healthy female volunteers.     

The antioxidant edaravone attenuates ER-stress-mediated cardiac apoptosis and dysfunction in rats with autoimmune myocarditis

Abstract

Experimental autoimmune myocarditis (EAM) is mediated by myocardial infiltration by myosin-specific T-cells secreting inflammatory cytokines. In this study, rat models of EAM were prepared by injection with porcine cardiac myosin. One week after immunization, edaravone was administered intraperitoneally at 3 or 10 mg/kg/day to rats for 2 weeks. Cardiac function was measured by haemodynamic and echocardiographic studies and TUNEL assay was performed. Left ventricular (LV) expression of NADPH oxidase sub-units (p47^phox and p67^phox), pro-inflammatory cytokines (TNF-α), endoplasmic reticulum (ER) stress signalling proteins (GRP78, caspase-12 and GADD153) and mitogen-activated protein kinase (MAPK) family proteins (phospho-p38 MAPK and phospho-JNK) were measured by western blotting. Edaravone improved LV function in a dose-dependent manner. Central venous pressure was significantly low and LV ejection fraction and fractional shortening was significantly high in edaravone groups compared with those in the vehicle group. In addition, edaravone treatment down-regulated LV expressions of p47^phox, TNF-α, GADD153, phospho-p38 MAPK and phospho-JNK. Furthermore, the LV expressions of p67^phox, GRP78, caspase-12 and TUNEL-positive cells of rats with EAM treated with edaravone were significantly low compared with those of the vehicle group. These findings suggest that edaravone ameliorated the progression of EAM by inhibiting oxidative and ER stress and, subsequently, cardiac apoptosis.     

Study on survival,growth, haematology and body composition of Cyprinus carpio under different acute and chronic salinity regimes

Pakistan’s most of the land is less productive or no productivity at all due to erosion and salinity of the soil, which can be utilized to develop fisheries. The project, “Survival, growth and body composition of Cyprinus carpio under different salinity regimes” was undertaken in two phases. In the first phase susceptibility of Cyprinus carpio at four salinity levels in triplicate within 0–10 g L⁻¹NaCl for 96 h in each aquarium was checked after one week acclamation at 0 g L⁻¹, 2 g L⁻¹ and 4 g L⁻¹ NaCl. LC₅₀ values varied from 7.67 to 10.65 g L⁻¹ after 96 h for C. carpio. Percentage mortality of the fish and important water quality parameters after every 12 h were observed for a period of 96-h. Probit analysis showed that 96-h LC₅₀ values ranged from 7.67 to 10.65 g L⁻¹. During experimental period aquaria water temperature ranged from 29.6 to 33.7 °C, pH values fluctuated between 7.8 and 9.7, Electrical conductivity values ranged from 2.40 to 20.13 dSm⁻¹ and Dissolved oxygen ranged between 2.23 and 10 mg L⁻¹. Sub-lethal salt concentration i.e. 0 g L⁻¹ to 3 g L⁻¹ NaCl upto 40 days showed that growth of C. carpio decreased with the increase of water salinity levels and ceased at 4 g L⁻¹ salinity and increase in salinity have negatively affected hematological parameters.     

Piwi positive cells that line the vasculature epithelium, underlie whole body regeneration in a basal chordate

The colonial tunicate Botrylloides leachi can regenerate functional adults from minute vasculature fragments, in a poorly understood phenomenon termed Whole Body Regeneration (WBR). Using Piwi expression (Bl-Piwi), blood cell labeling and electron microscopy, we show that WBR develops through activation, mobilization and expansion of ‘dormant’ cells which normally line the internal vasculature epithelium of blood vessels. Following a mechanical insult, these cells express Bl-Piwi de novo, change morphology and invade niches of the vasculature lumen, where they proliferate and differentiate, regenerating a functional organism. Mitomycin C treatments and siRNA knockdown of Bl-Piwi result in deficient cells incapable of expanding or differentiating and to subsequent regeneration arrest. Last, we find similar transient mobilization of Piwi⁺ cells recurring every week, as part of normal colony development, and also during acute environmental stress. This recurrent activation of Piwi⁺ cells in response to developmental, physiological and environmental insults may have enabled the adaptation of colonial tunicates to the imposed varied conditions in the marine, shallow water environment.     

Effects of BMP-2 compound with fibrin on osteoporotic vertebral fracture healing in rats

Objectives:To investigate the effects of bone morphogenetic protein-2 (BMP-2) compound with fibrin on osteoporotic vertebral fracture healing in rats.Methods:For the present study 160 Specific-Pathogen Free 32-week-old female Sprague-Dawley rats were used. 120 rats were randomly divided in three groups (experimental, model and sham operation group- n=40 per group) and were ovariectomized to establish the osteoporosis model. 40 rats served as a control group without treatment. The expression of BMP-2 in the fracture zone at the 4^th, 6^th, 8^th, and 12^th weeks was detected by qRT-PCR. The expression of BALP and CTX-I in serum at the 12^th week was detected by Elisa.Results:At week 8, the morphology of the sham operation group was the same and the fracture healing occurred more slowly than in the other groups. At week 12, the expression of BMP-2 in the model group was significantly higher than that in the other three groups (p<0.05). At week 12, the maximum load, maximum strain, and elastic modulus of model group were significantly lower than those of the other three groups.Conclusions:BMP-2 compound with fibrin can enhance the timing and quality of bone fracture healing in rats.     

Daily Variations in the Responses to Papaverine but not Theophylline in Rat Aorta

The presence of temporal patterns in the responses of vascular smooth muscle to relaxing agents have been known. The aim of this study was to examine the variations in the in vitro sensitivity of rat aorta to papaverine and theophylline in both endothelium-intact and -denuded rings. The animals were synchronized with a 12-h light: 12-h dark schedule. Rat thoracic aorta rings were obtained from animals sacrificed at six different times (09:00, 13:00, 17:00, 21:00, 01:00 and 05:00) of the day. Tissues were precontracted submaximally with phenylephrine and then were relaxed by papaverine (10^?8 ? 10^?4 M) and theophylline (10^?7 ? 10^?3 M) applied cumulatively. The responses to most of the concentrations and EC₅₀ values for papaverine demonstrated an ultradian rhythm with an 8 h periodicity. Time-dependent variations were not found in the responses to theophylline. The results are discussed in relation with the time-dependent differences in signal transducing mechanisms.     

Changes in activity,spatial pattern and social behavior in calves after grouping

The behavior of 2 groups of 6 calves that were managed in 2 different ways were observed, and the development of their behavioral patterns, spatial patterns and social behavior in the groups were discussed, together with the effect on these processes of their social experience before grouping.Twelve 5-month-old Holstein steers, reared alone from one week of age, were divided into 2 groups of 6 calves each; Groups A and B. Calves in Group A were penned together in a 14.4 × 28.8 m pen, and the position and behavior of individuals was observed throughout a continuous 153-h period at 15-min intervals. Aggressive behavior between calves was also recorded. The position of calves was recorded as coordinates on a grid on the pen floor. There were 8 × 16 squares in the grid, and each square measured 1.8 × 1.8 m.Group B was further divided into 3 groups of 2 calves each, which were kept in 9.6 × 14.4 m pens. The pair-mates were changed every 3 days in order to combine all possible pairs in the group. After this treatment, calves were all housed together in a 14.4 × 28.8 m pen, and observed using the same methods and for the same duration as Group A.The lying pattern of Group A was not diurnal until the latter half of the observation, while calves in Group B showed clear-cut diurnal variations from the initial stages. Concerning the spatial patterns; the mean distance to nearest neighbor in Group A tended to decrease and become stable. The ratio between the mean distance to nearest neighbor and the distance expected in random distribution also declined from 0.7 to a significant aggregation of 0.5 (P < 0.05). For Group B, a similar change in the mean distance to the nearest neighbor was observed, although this showed significant aggregation from the first 24-h period (P < 0.05). The mean area occupied by a group in each 24-h period, for both groups, declined for 72 h after grouping and became stable at around 18 m², for both groups, thereafter. The frequency of aggression in Group A declined from 23 times in the first 24-h period to 9 times at the end of the observation period. For Group B, the aggression frequency was 53 times in the first 24-h period, thereafter decreasing rapidly.     

Receptor-mediated uptake of homologous low-density lipoproteins by isolated liver parenchymal cells of fetal rats

Summary The binding and uptake of gold-labeled homologous, apolipoprotein E-free low-density lipoproteins (LDL) by isolated fetal rat liver parenchymal cells in suspension were studied ultrastructurally and morphometrically. Binding experiments using ¹²⁵I-labeled LDL were also performed. After a 2-h preincubation in a lipoprotein-free medium and a subsequent 1-h postincubation in the presence of LDL-gold, fetal liver parenchymal cells exhibit a binding of 248±17 gold conjugates/100 m plasma membrane and an uptake of 235±17 gold conjugates/100 m² cytoplasm. Compared with values obtained from freshly isolated nonpreincubated cells, these data correspond to a 15-fold and an 18-fold increase in total binding and uptake of LDL-gold, respectively. Competition experiments reveal that this increase is mainly a result of a 23-fold stimulation of specific binding and a 44-fold stimulation of receptor-mediated uptake of LDL-gold. The ¹²⁵I-LDL binding experiments give a K_d value of 6.3×10^-8 M and a maximum binding capacity of 17.3 fmol LDL/10⁶ cells. Our data provide evidence, further to our in vivo studies, that fetal rat liver parenchymal cells possess high-affinity binding sites for native homologous apolipoprotein E-free LDL. These sites may correspond to B, E receptors of adult rat liver parenchymal cells.     

<Superscript>177</Superscript>Lu-DOTA-coupled minigastrin peptides: promising theranostic agents in neuroendocrine cancers

Treatment with radionuclide labeled regulatory peptides is a promising tool in the management of patients with inoperable receptor positive neuroendocrine tumors. Peptide receptor lutetium-177 radionuclide therapy currently has gained ample attention due to high specific accumulation of regulatory peptides at tumor cell surface and promising characteristics of β- and γ-energy photons of lutetium-177 radionuclide. In this study gastrin peptides analogues were labeled with lutetium-177 by subsequent mixing of ¹⁷⁷LuCl₃ (~?185 MBq), ammonium acetate buffer of 5 pH, gentistic acid, aqueous solution of gastrin peptide analogues (1 mg/mL) and heating the reaction mixture at 98 °C which resulted in high radiochemical yield (>?96%). Chromatographic analysis was carried out to analyze the radiochemical purity. The shelf life and serum stability results showed the labeled peptides are sufficiently stable up to 4-h. Glomerular filtration rate study results showed moderate filtration through kidneys. The GFR values of ¹⁷⁷Lu-MGCL2 and ¹⁷⁷Lu-MGCL4 was noted 48 mL/min and 45 mL/min, respectively. Biodistribution and scintigraphy study using rat and rabbit models showed minimal non-target accumulation, moderate uptake by liver and kidneys. The promising radiochemical yield, stability, GFR values and biodistribution results of ¹⁷⁷Lu-MGCL2 & 4 indicate, the agents can be tested clinically for PRRT procedures.     

Clinical experience with two simple methods of measurement of 99mTc-methylene diphosphonate body retention in bone disease.

Whole-body retention (WBR) of 99mTc-methylene diphosphonate was measured in 15 control subjects and in 99 patients with various metabolic bone diseases (osteoporosis, Paget's disease of the bone, hyperparathyroidism). In all the subjects WBR was measured indirectly from urinary excretion and in 30 it was also measured directly using a simple detector. Mean WBR values did not differ between control subjects and osteoporotic patients, whereas patients with Paget's disease and patients with hyperparathyroidism had significantly higher mean WBR levels than the controls. However, overlapping of results among the groups was high, and consequently the diagnostic value in the individual patients was low. In the patients whose WBR levels were measured simultaneously by both the direct and the indirect urinary method, the results were essentially the same, provided urine losers were excluded. Comparison of the two detection methods showed that urine loss occurred frequently and this was probably partly responsible for the low diagnostic sensitivity of the WBR. When WBR was measured at different time intervals in untreated individuals, the values remained markedly constant. The method would probably be of value in monitoring treatments influencing bone turnover in metabolic bone diseases.     

CREB activation in the rapid, intermediate, and delayed ischemic preconditioning against hypoxic–ischemia in neonatal rat

Ischemic preconditioning (IP) is a defense program in which exposure to sublethal ischemia followed by a period of reperfusion results in subsequent resistance to severe ischemic insults. Very few in vivo IP models have been established for neonatal brain. We examined whether rapid, intermediate, and delayed IP against hypoxic–ischemia (HI) could be induced in neonatal brain, and if so, whether the IP involved phosphorylation of cAMP response element-binding protein (pCREB) after HI. Postnatal day 7 rat pups were subjected to HI at 2 h (2-h IP), 6 h (6-h IP), or 22 h (22-h IP) after IP. We found all three IP groups had significantly reduced neuronal damage and TUNEL-(+) cells 24 h post-HI than no-IP group. Compared with control, the no-IP group had significant decreases of pCREB and mitochondria Bcl-2 levels in the ipsilateral cortex 24 h post-HI. In contrast, the three IP groups had increased pCREB and mitochondria Bcl-2 levels, and significant differences were found between three IP and no-IP groups. The increases of cleavage of caspase-3 and poly (ADP-ribose) polymerase and of cells with nuclear apoptosis inducing factor post-HI in no-IP group were all significantly reduced in three IP groups. The increases of caspase-3 and calpain-mediated proteolysis of α-spectrin post-HI were significantly reduced only in 22-h IP group. Furthermore, all three IP groups had long-term neuroprotection at behavioral and pathological levels compared with no-IP group. In conclusion, IP, rapid, intermediate, or delayed, in neonatal rat brain activates CREB, up-regulates Bcl-2, induces extensive brakes on caspase-dependent and -independent apoptosis after HI, and provides long-term neuroprotection.