Inactivation of Photosensitizing Merocyanine Dyes by Plasma, Serum and Serum Components

Abstract— Merocyanine dyes with an oxygen in the electron donor heterocycle were rapidly degraded by plasma, serum and serum components. Replacement of the oxygen by a sulfur or selenium atom rendered the dyes refractory to degradation. The degradation of labile merocyanine dyes was temperature dependent and oxygen independent. The plasma component that was responsible for the degradation of merocyanine dyes was sensitive to heat and detergent, suggesting an enzymatic process. The identification of the structural requirements for sensitivity/resistance to degradation provides the experimenter with a simple means to manipulate the stability of mer-ocyanines in high serum or plasma environments and may expand the clinical utility of merocyanine photosen-sitizers beyond their traditional role in the extracorporeal purging of bone marrow grafts.     

Evaluation by using radionuclide uptake of bone in Paget's disease of bone: special reference to treatment with calcitonin

Bone scintigraphy using 99mTc-MDP was performed on 2 patients with Paget's disease of bone before and after the treatment with a synthetic eel calcitonin analogue [Asu1,7)-eel calcitonin, ECT] at a dose of 40 U per day. All pagetic lesions showed markedly the increased accumulation of the radionuclide. The uptake ratio, defined as the count rate of 99mTc-MDP over each bone lesion to that over the control bone, was calculated. The response to the calcitonin therapy was evaluated with the uptake ratio of the radionuclide. The uptake ratio decreased markedly within the first 3 months of the treatment, in association with a palliation of bone pain, while the serum alkaline phosphatase activities which had been within the normal range or slightly high before the treatment did not show any significant change or did not reflect a clinical feature (e.g. bone pain) with the treatment. Thus, the uptake ratio on the bone scintigram seemed to offer the most sensitive and most reliable information for the evaluation of calcitonin treatment of Paget's disease of bone.     

Bone marrow scintigraphy with 111In-chloride--a clinical value for the hematological diseases

Bone marrow scintigraphy with indium chloride (111In) was performed in fifty-one patients with the hematological diseases. The results of the investigation were that 1. in all patients, as well as in patients with aplastic anemia, no correlation was there between the degree of the indium chloride accumulation and peripheral blood counts, 2. in patients with aplastic anemia and pure red cell aplasia (PRCA) a tendency to reduction in uptake of indium chloride in bone marrow, 3. in patients with these two good correlation between the degree of indium chloride accumulation and histology of the erythroid bone marrow, but in patients with chronic myelocytic leukemia (CML) and atypical leukemia no correlation between the two, so it seemed unlikely that indium chloride should reflect the effective production of erythrocytes, 4. four patients with leukemia were studied with indium chloride bone marrow imaging two times to evaluate their responses to chemotherapy, and peripheral expansion was no change or reduced in two patients with acute myelocytic leukemia (AML) and one patient with acute lymphocytic leukemia (ALL) who obtained complete remission, but on the other hand, it enlarged in one patient with acute myelocytic leukemia who obtained partial remission, and 5. in two patients with chronic myelocytic leukemia it enlarged up to the ankle joints, which was considerably specific.     

再障头发、血清和骨髓组织钴、锂元素含量的变化

采用原子吸收光谱技术测定再障４３例患者头发、血清和骨髓组织的钴、锂元素含量，并与急性白血病２０例、缺铁性贫血１０例和正常人４４例作对照比较，再障头发、骨髓钴、锂和血清钴含量均低于正常人，Ｐ＜０．０５，而与急性白血病和缺铁性贫血之间，以及各种类型再障之间则无差异性．再障头发锂含量与血红蛋白、红细胞数呈负相关，Ｐ＜０．０５，说明再障确实存在着钴、锂元素代谢异常．     

The relation of various metabolic bone markers in renal hemodialysis patients classified by serum estradiol levels

The bone metabolic markers showed the significant differences in age and sex of the hemodialysis patients with renal insufficiency. It is likely that the evaluations of bone change and biochemical metabolic markers should be based on serum estradiol level, sex and age of the renal hemodialysis patients.     

Ion-Paired Liquid Chromatographic Determination of Cefazolin in Canine Serum and Tissues

Abstract

Cefazolin is commonly used as a prophylactic antibiotic in dogs undergoing total hip arthroplasty.

A sensitive high-performance liquid chromatographic method was developed for the determination of cefazolin in canine serum and tissues. The tissues were those in contact with the hip prothesis; namely, the coxofemoral joint capsule, cancellous bone from the acetabulum and bone marrow from the femoral canal.

The method involved filtration of diluted serum or tissue extracted with ethanol:acetonitrile:water (40:10:50) through a 30,000 molecular weight cut-off filter. Separation of cefazolin from other components was by ion-paired (dodecanosulfonate) high performance liquid chromatography using a reversed-phase column eluted with acetonitrile/water solution. The ultraviolet absorbance of the column effluent was monitored at 230 nm. Recovery of cefazolin spiked at 10 μg/ml from serum was 89.8% with a coefficient of variation (CV) of 5.3% (n=10). Recovery of cefazolin spiked at 5 μg/ml from tissue extracts of joint capsule, cancellous bone and bone marrow was: 93.9%, 98.2%, and 104.2% respectively, with a CV of 6.7%, 10.2% and 10.6% respectively (n=10). A correlation coefficient of 0.9999 occurred with cefazolin in aqueous solution (n=5). The limit of detection for cefazolin was approximately 10 ng/ml of serum or tissue extract.     

再障患者锗元素检测和粒单祖细胞培养的临床意义探讨

采用原子吸收光谱技术测定再障患者４３例头发，血清和骨髓组织的锗元素含量，并与急性白血病２０例，缺铁性贫血１０例和正常人４４例作对照比较，再障，急性白血病和缺铁性贫血组头发，血清锗含量均低低于正常人，Ｐ〈０．０５；再障骨髓锗含量同时也低于正常人，Ｐ〈０．０５；再障骨髓锗含量同时也低于正常人，Ｐ〈０．０５；原发生再障发锗含量低于继发性再障，Ｐ〈０．０５。再障骨髓ＧＭ－ＣＦＵ的数量则显著低于正常人，Ｐ〉     

多发性骨髓瘤的骨髓形态学分析与临床应用

目的研究多发性骨髓瘤的骨髓形态学特征,探讨骨髓形态学在多发性骨髓瘤诊断中的临床应用价值。方法对广东医学院附属医院63例多发性骨髓瘤病人进行骨髓涂片及细胞染色和进行检验诊断分析。结果 94%的患者骨髓增生度都达到增生活跃以上,只有6%的是增生低下或极度低下,瘤细胞数量﹥15%,根据形态可分为四类;病人骨髓其他系包括巨核系、粒系、红系和淋巴系细胞都不同程度减少和受抑制。结论骨髓形态学分析是多发性骨髓瘤诊断和治疗疗效监测具有重要的临床应用价值,具有广泛开展的必要性和意义。     

Inhibition of renal permeability towards albumin: a new function of apolipoproteins with possible pathogenetic relevance in focal glomerulosclerosis.

Focal segmental glomerulosclerosis (FSGS) is a degenerative renal disease characterized by the accumulation of extracellular matrix and lipids within the glomerular tuft. It has been proposed that an abnormal renal permeabilization towards proteins induced by a putative plasma factor is, in some way, involved in the pathogenesis of the disease. In this paper, we measured the plasma permeability activity (Palb) in several sera of patients with FSGS and found a mean activity of 0.82+/-0.03 which means a marked increase compared to a mean Palb of 0.16+/-0.03 in normal controls. Coincubation of FSGS and normal serum reduced the permeability activity within the normal range; normal serum added to the incubation medium after the glomeruli had already been exposed to the FSGS serum had no effect, suggesting the presence of inhibitory substances with a direct effect on a circulating substrate. Finally, the antipermeability activity was retained when heated to 60 degrees C but not to 100 degrees C. By serial fractionations of normal serum and reported activity measurements at each step, five natural occurring inhibitors of albumin permeabilization were purified and characterized by matrix assisted laser desorption/ionization-mass spectrometry (MALDI-MS), as components of apolipoproteins (apo) (apo E2 and E4, apo L, the high Mr apo J and a 28 kDa fragment of apo A-IV). Coincubation of each apolipoprotein with FSGS serum inhibited permeability, but only apo J and apo E2 and E4 were found to be crucial for the process. In conclusion, we have purified from normal serum five inhibitors of permeability induced by FSGS serum, all corresponding to apolipoproteins. An imbalance between permeability factors and apolipoproteins may play a pathogenetic role in FSGS.     

Radiotracers for Bone Marrow Infection Imaging

Introduction: Radiotracers are widely used in medical imaging, using techniques of gamma-camera imaging (scintigraphy and SPECT) or positron emission tomography (PET). In bone marrow infection, there is no single routine test available that can detect infection with sufficiently high diagnostic accuracy. Here, we review radiotracers used for imaging of bone marrow infection, also known as osteomyelitis, with a focus on why these molecules are relevant for the task, based on their physiological uptake mechanisms. The review comprises [⁶⁷Ga]Ga-citrate, radiolabelled leukocytes, radiolabelled nanocolloids (bone marrow) and radiolabelled phosphonates (bone structure), and [¹⁸F]FDG as established radiotracers for bone marrow infection imaging. Tracers that are under development or testing for this purpose include [⁶⁸Ga]Ga-citrate, [¹⁸F]FDG, [¹⁸F]FDS and other non-glucose sugar analogues, [¹⁵O]water, [¹¹C]methionine, [¹¹C]donepezil, [^99mTc]Tc-IL-8, [⁶⁸Ga]Ga-Siglec-9, phage-display selected peptides, and the antimicrobial peptide [^99mTc]Tc-UBI_29-41 or [⁶⁸Ga]Ga-NOTA-UBI_29-41. Conclusion: Molecular radiotracers allow studies of physiological processes such as infection. None of the reviewed molecules are ideal for the imaging of infections, whether bone marrow or otherwise, but each can give information about a separate aspect such as physiology or biochemistry. Knowledge of uptake mechanisms, pitfalls, and challenges is useful in both the use and development of medically relevant radioactive tracers.     

Quantification of mitoxantrone in bone marrow by high-performance liquid chromatography with electrochemical detection

An analytical method for the determination of mitoxantrone in bone marrow was developed using high-performance liquid chromatography with electrochemical detection. The extraction procedure was optimized by investigating several factors which potentially could influence the recovery of mitoxantrone from bone marrow cells. The mean recovery of mitoxantrone from rat bone marrow was found to be 81.7% with a coefficient of variation 3.8%. High-performance liquid chromatography was carried out to quantitate mitoxantrone using ametantrone as internal standard. The detection limit of our analytical method amounts to 100 pg on-column, corresponding to 1 ng/ml of cell suspension containing 2 x 10(7) cells and a day-to-day variation of maximally 8%. Storage of bone marrow samples, containing mitoxantrone, for one to fourteen days resulted in a mean recovery of 94%, as compared to freshly analysed samples. Subsequently we studied the pharmacokinetics of mitoxantrone in rat bone marrow. It appeared that after an intravenous bolus injection of mitoxantrone (2.5 mg/kg) in rats, the drug accumulated in the femoral bone marrow for about four days, and thereafter gradually declined.     

再障头发、血清和骨髓组织锌、硒元素测定及临床意义探讨

对２５例再障患者头发、血清和骨髓组织中锌、硒含量进行测定，并与２０例健康人作比较，再障头发、血清和骨髓内锌、硒含量均明显低于正常人，Ｐ＜０．０１；有感染的１３例患者血清锌低于正常人，Ｐ＜０．０１；未有感染的病人血清锌与正常组无差异，Ｐ＞０．０５．提示再障患者存在着锌、硒元素代谢异常，可能是再障发病机理的因素之一．     

Measurement of 24-hr whole-body retention of 99mTc-MDP with a thyroid uptake probe (author's transl)

A new method for measurement of 24-hr whole-body retention (WBR) of 99mTc-MDP, using a thyroid uptake probe was established and its clinical significance was evaluated in various bone diseases. (1) Reproducibility of 24-hr WBR in 9 patients was very good. Correlation coefficient was 0.997 and coefficient of variability was only 1.83%. (2) Radiochemical purity of 99mTc-MDP was 97.8 +/- 0.7% (n = 5), indicating no significant inter-lot variations. (3) 24-hr WBR of normal adult males (n = 5) was 30.0 +/- 4.9%, which was significantly elevated compared to the reported 99mTc-HEDP WBR of 19.2 +/- 1.7%. Whole-body retentions of chronic renal failure, metastatic bone disease and hyperthyroidism groups were significantly elevated compared to that of the normal group. However, WBR of steroid-induced osteoporotic group was significantly decreased. Based on these results, this thyroid uptake probe method was simple, reproducible and accurate to measure 24-hr WBR of 99mTc-MDP. Quantification of WBR of 99mTc-MDP was of great clinical value to diagnose metabolic bone disease and to follow-up metabolic and metastatic bone diseases.     

Synthesis and biological evaluation of a novel <Superscript>99m</Superscript>Tc complex of HYNIC-conjugated aminomethylenediphosphonate as a potential bone imaging agent

A conjugate of 6-hydrazinopyridine-3-carboxylic acid (HYNIC) with aminomethylenediphosphonic acid (AMDP) was synthesized through a multiple-step reaction. HYNIC–AMDP could be labeled easily and efficiently with ^99mTc using N-(2-hydroxy-1,1-bis(hydroxymethyl)ethyl)glycine (tricine) as coligand to form the ^99mTc–HYNIC–AMDP complex in high yield (> 95%). Its partition coefficient indicated that it was a good hydrophilic complex. The biodistribution studies of ^99mTc–HYNIC–AMDP in normal ICR mice showed that this complex had high bone uptake and low or negligible accumulation in non-target organs. As compared with ^99mTc–MDP, ^99mTc–HYNIC–AMDP had a higher bone uptake and the ratios of bone/blood and bone/muscle at early time after injection, suggesting that it could be potentially useful for bone imaging at an earlier time after injection according to further investigations of the biological behavior of this complex.     

Trace aluminum determination and sampling problems of archeological bone employing destructive neutron activation analysis

A destructive neutron activation analysis procedure was developed for determining trace aluminum content in bone. It was found that soil contamination can influence the aluninum bone levels in prehistoric bone specimens. These maximum aluninum content values for prehistoric bone are larger than those of modern bone and comparable to aluminum levels present in bone from renal patients.     

代谢组学法分析骨髓抑制模型小鼠血清小分子代谢产物

骨髓抑制是恶性肿瘤患者化疗后常见的症状,研究骨髓抑制造成体内代谢产物的变化可以为诊断骨髓抑制病症发展状况及采用药物治疗提供思路。采用雄性BalB/C小鼠腹腔注射环磷酰胺建立骨髓抑制模型,通过气相色谱-质谱(GC-MS)对正常与造模小鼠血液代谢产物进行指纹图谱分析,然后采用主成分分析(PCA)和正交偏最小方差判别分析(OPLS-DA)对代谢组学数据进行多维统计分析。与对照组相比,骨髓抑制模型小鼠血浆中潜在的差异表达代谢标志物有15个,其中葡萄糖-1-磷酸、对硝基苯酚、乙酰苯胺、可的松、烟酰胺、马钱苷、咖啡酸、亚油酸和油酸这9种物质的表达差异有统计学意义(P0.05)。结果表明,代谢产物可作为骨髓抑制研究中的重要标记物,有助于揭示化疗所致骨髓抑制的发病机制,判断疾病发展阶段以及后续治疗手段的有效性。     

Pharmacokinetics and tissue distribution of the antileukaemic organoarsenicals arsthinol and melarsoprol in mice

Melarsoprol and arsthinol have been shown to be effective on various leukaemia cell lines. Nevertheless, the tissue distribution of these compounds remains a key point since the bone marrow is considered as the site of action and the central nervous system as the site of the main toxicity. In this study, we have determined the exposure of each organ (blood, liver, bone marrow and brain) to arsenic, irrespectively of the exact nature of arsenic species contained by the organ.In the bone marrow, arsenic concentrations were very high, especially that of melarsoprol. However, the lower ability of arsthinol to concentrate in the bone marrow could be compensated by its higher antileukaemic activity.The brain concentrations were high, although lower than in the bone marrow; this fact is in very good accordance with the observation that the brain is mainly involved in the acute toxicity of trivalent organoarsenicals.     

High-performance liquid chromatographic-mass spectrometric assay of busulfan in serum and cerebrospinal fluid.

A liquid chromatographic-mass spectrometric method has been developed to determine busulfan concentrations in the cerebrospinal fluid and serum of some children undergoing bone marrow autotransplantation. After two liquid-liquid extraction steps with dichloromethane on a biological matrix, the separation of busulfan was carried out by isocratic reversed-phase chromatography. The mass spectrometric system was operated in electron-impact mode. Principal ions at m/z 175, 111 and 79 were observed for busulfan, but only m/z 175 was chosen for the quantification of the analyte. The retention time of busulfan was 2.5 min. The detection limit of 100 ng/ml allowed the determination of cerebrospinal fluid and serum busulfan concentrations during the four days of high-dose (1 mg/kg) treatment prior to autotransplantation in five child patients.     

白血病骨髓移植患者口腔黏膜病与微量元素锌的关系

为研究白血病骨髓移植患者全血微量元素锌与口腔黏膜病的关系，用原子吸收光谱法检测了正常对照组与白血病骨髓移患者预处理前及移植后骨髓空虚期全血微量元素锌的含量，并观察了白血病骨髓移植患者空黏膜病变情况。结果表明，正常对照组全血锌浓度与白血病骨髓移植患者预处理前全血锌浓度比较，差异无显著性（P＞0．05），而预处理前全血锌浓度与骨髓空虚期全血锌浓度有显著性差异（P＜0．01），白血病缓解后（预处理前）血锌接受正常水平，无口腔溃疡发生，而白血病骨髓移植患者处理后骨髓空虚期全血锌含量显著降低，并出现不同程度的口腔黏膜病变，说明预处理影响微量元素锌的代谢，微量元素锌的减少与白血病骨髓移植患者移植患者口腔黏膜病的发生有关。     

Activation of 4-1BB signaling in bone marrow stromal cells triggers bone loss via the p-38 MAPK-DKK1 axis in aged mice

Senile osteoporosis can cause bone fragility and increased fracture risks and has been one of the most prevalent and severe diseases affecting the elderly population. Bone formation depends on the proper osteogenic differentiation of bone marrow stromal cells (BMSCs) in the bone marrow microenvironment, which is generated by the functional relationship among different cell types in the bone marrow. With aging, bone marrow provides signals that repress osteogenesis. Finding the signals that oppose BMSC osteogenic differentiation from the bone marrow microenvironment and identifying the abnormal changes in BMSCs with aging are key to elucidating the mechanisms of senile osteoporosis. In a pilot experiment, we found that 4-1BBL and 4-1BB were more abundant in bone marrow from aged (18-month-old) mice than young (6-month-old) mice. Meanwhile, significant bone loss was observed in aged mice compared with young mice. However, very little data have been generated regarding whether high-level 4-1BB/4-1BBL in bone marrow was associated with bone loss in aged mice. In the current study, we found upregulation of 4-1BB in the BMSCs of aged mice, which resulted in the attenuation of the osteogenic differentiation potential of BMSCs from aged mice via the p38 MAPK-Dkk1 pathway. More importantly, bone loss of aged mice could be rescued through the blockade of 4-1BB signaling in vivo. Our study will benefit not only our understanding of the pathogenesis of age-related trabecular bone loss but also the search for new targets to treat senile osteoporosis.Subject terms: Stem-cell differentiation, Mesenchymal stem cells, Ageing