Comparative Cell Shape and Diffusional Water Permeability of Red Blood Cells from Indian Elephant ( Elephas maximus ) and Man ( Homo sapiens )

Red blood cells (RBC) from an Indian elephant (Elephas maximus) were studied by light microscopy (LM), scanning electron microscopy (SEM) and a new nuclear magnetic resonance (NMR) ‘imaging’ method based on the translational diffusion of water, NMR q-space analysis. Also, the transmembrane diffusional permeability, P _d of water in RBC was measured by using a Mn²⁺-doping NMR technique, taking human RBC as a reference. The main diameter of the elephant RBC was measured as 9.3 ± 0.7 μm by LM, 9.3 ± 0.7 μm by ‘shrinkage-corrected’ SEM, and 9.3 ± 0.4 μm by q-space anlaysis. The value is ∼1.4 μm larger than that for the human RBC. The values of P _d were, in the case of elephant RBC, 3.2 × 10⁻³ cm/s at 25 °C, 3.9 × 10⁻³ cm/s at 30 °C, 5.2 × 10⁻³ cm/s at 37 °C and 6.5 × 10⁻³ cm/s at 42 °C; all values were significantly lower than the corresponding values of P _d for human RBC, namely 4.3 × 10⁻³ cm/s at 25 °C, 5.2 × 10⁻³ cm/s at 30 °C, 6.1 × 10⁻³ cm/s at 37 °C, 7.8 × 10⁻³ cm/s at 42°C. The maximal inhibition of P _d (56%) was reached in 30 min at 37 °C with 2 mm p-chloromercuribenzene sulphonate (PCMBS) for both species of RBC. The basal permeability to water at 37 °C was estimated to be 2.3 × 10⁻³ cm/s for elephant and 2.6 × 10⁻³ cm/s for human RBC. The values of the activation energy for water permeability (E _a,d ) was significantly higher for elephant RBC (31.9 kJ/mol) than for human RBC (25.9 kJ/mol). This indicated that features other than the number of transporters per cell are likely to be important in defining the differences in water permeability in the RBC from the two species.     

Comparative Nuclear Magnetic Resonance Studies of Diffusional Water Permeability of Red Blood Cells from Different Species. XI. Horses Introduced to Australia and European Horses ( Equus caballus )

The diffusional water permeability (P _d) of red blood cells (RBCs) from horses introduced to Australia and from European horses was measured by a Mn²⁺ doping nuclear magnetic resonance (NMR) technique. The values of P _d were ∼3.5 × 10⁻³ cm/s at 25°C, 4.1 × 10⁻³ cm/s at 30°C, 5.6 × 10⁻³ cm/s at 37°C and 6.3 × 10⁻³ cm/s at 42°C with no significant differences between the two strains of horse. Systematic studies on the effect of p-chloromercuribenzene sulphonate (PCMBS) on water diffusion indicated that the maximal inhibition (60%) was reached in 60 min at 22°C with 1 mm PCMBS. The basal permeability to water was estimated at 1.2 × 10⁻³ cm/s at 25°C, 1.7 × 10⁻³ cm/s at 30°C, 2.0 × 10⁻³ cm/s at 37°C and 2.7 × 10⁻³ cm/s at 42°C. The activation energy (E _a,d) of water diffusion was ∼25 kJ/mol and increased to ∼39 kJ/mol after incubation with PCMBS, in conditions of maximal inhibition of water diffusion. The membrane protein electrophoretic pattern of horse RBCs was compared with its human counterpart. The horse RBC membranes had lower amounts of the proteins migrating as bands 4.1 and 4.2 and higher amounts of the protein migrating as band 4.9, and band 6 (glyceraldehyde-3-phosphate dehydrogenase) was undetectable. A noteworthy feature was the appearance of considerable differences in protein migration distances in the region of bands 7 and 9, between horse and human membranes.     

Comparative Nuclear Magnetic Resonance Studies of Diffusional Water Permeability of Red Blood Cells from Different Species. XII. Dog (<Emphasis Type="BoldItalic">Canis familiaris</Emphasis>) and Cat (<Emphasis Type="BoldItalic">Felis domestica</Emphasis>)

The diffusional water permeability (P _d ) of dog and cat red blood cell (RBC) membrane has been monitored by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition with p-chloromercuribenzene sulphonate (PCMBS). The values of P _d were in the case of cat RBC ∼3.0 × 10⁻³ cm/s at 15 °C, 3.5 × 10⁻³ cm/s at 20 °C, 4.2 × 10⁻³ cm/s at 25 °C, 4.4 × 10⁻³ cm/s at 30 °C and 5.9 × 10⁻³ cm/s at 38 °C. In case of dog RBC the values of P _d were higher ∼3.8 × 10⁻³ cm/s at 15 °C, 4.6 × 10⁻³ cm/s at 20 °C, 5.0 × 10⁻³ cm/s at 25 °C, 5.9 × 10⁻³ cm/s at 30 °C and 7.9 × 10⁻³ cm/s at 37 °C. Systematic studies of the effect of PCMBS on water diffusion indicated that in the case of dog RBCs the maximal inhibition was reached in 15–30 min with 1 mm PCMBS, whereas in the case of cat RBCs in 60 min with 1 mm PCMBS or in 30 min with 2 mm PCMBS. The values of maximal inhibition in the case of cat RBC were in the range of 55–60% at 15 °C, 60–68% at 20 °C and 25 °C, 50–60% at 30 °C and 50–55% at 37 °C. In the case of dog RBC the corresponding values were higher, 75–80% at 15 °C, 70–80% at 20 °C and 25 °C, 65–70% at 30 °C and 55–60% at 37 °C. The basal permeability to water was estimated to be ∼1 × 10⁻³ cm/s −2 × 10⁻³ cm/s in the range of temperatures of 25–37 °C. The activation energy of water diffusion E _a,d was ∼19 kJ/mol for the dog RBC and ∼23 kJ/mol for the cat RBC. After incubation with PCMBS the values of E _a,d increased, reaching 40 kJ/mol in conditions of maximal inhibition of water exchange. The membrane polypeptide electrophoretic pattern of dog and cat RBCs has been compared with its human counterpart. Dog and cat RBCs contained higher amounts of spectrin (band 1 and 2) and lower amounts of bands 4.4, 4.2, band 5 and band 7 compared to human RBCs. Band 4.9 was decreased only in the cat RBCs, whereas band 6 was decreased only in the dog RBCs. Correspondence and offprint requests to: Gheorghe Benga, Department of Cell and Molecular Biology, ‘Iuliu Haţieganu’ University of Medicine and Pharmacy, 6 Pasteur St, 3400 Cluj-Napoca, Romania. Tel:/Fax: 40–64–194373; e-mail: GBenga@personal.ro; gbenga@umfcluj.ro     

Controlling Autoimmunity: Lessons from Responses to Red Blood Cells

Current treatments for autoimmune diseases, which rely on inducing generalised immunosuppression, are not only frequently ineffective, but also associated with serious side effects. There is, therefore, a pressing need to understand the mechanisms that normally prevent autoimmune damage, and how they can be restored. A key advance in recent years has been the demonstration that autoreactive helper T- (Th-) cells are pivotal in the development of animal models of autoimmunity, and therefore provide a target for specific immunotherapy. This paper reviews the role of Th-cell responses against red blood cells (RBC) in autoimmune haemolytic anaemia (AIHA), the mechanisms by which such responses are prevented in health, and how this knowledge can be exploited in the future to treat the disease.     

Comparative NMR studies of diffusional water permeability of red blood cells from different species. X. Camel ( Camelus dromedarius ) and alpaca ( Lama pacos )

The diffusional water permeability (P _d) of camel and alpaca red blood cells (RBCs) was measured by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition withp-chloromercuribenzene sulphonate (PCMBS). The values ofP _d were, in the case of alpaca RBC≈4.6×10⁻³ cm/s at 25°C, 5.4×10⁻³ cm/s at 30°C, 6.6×10⁻³ cm/s at 37°C and 7.7×10⁻³ cm/s at 42°C. In case of camel RBC the values ofP _d where ≈4.2×10⁻³ cm/s and 9.0×10⁻³ cm/s at 42°C. Systematic studies on the effects of PCMBS on water diffusion in camel RBC indicated that the maximal inhibition was reached in 45 min with 1–2 mm PCMBS. The values of maximal inhibition were around 47% at 25°C and 68% at 30°C for alpaca RBC and around 62% at 25°C and 56% at 37°C for camel RBC. The basal permeability to water of alpaca RBC was estimated at around 2.6×10⁻³ cm/s at 25°C, 1.7×10⁻³ cm/s at 30°C and of camel RBC as 1.8×10⁻³ cm/s at 25°C and 3.0×10⁻³ cm/s at 37°C. The values of the activation energy of water diffusion (E _{a, d}) were around 23 kJ/mol for camel and 34 kJ/mol for alpaca RBC. This suggests that in addition to the number of transport channels other features of the pathways might be important for defining the temperature dependence of the water permeability.     

Oxidant Defence in the Red Blood Cells of the Greyheaded Flying Fox, Pteropus poliocephalus

The red blood cells (RBC) of bats (Order Chiroptera) are morphologically similar to those of other mammals, but the suborder Megachiroptera are unique in responding to very high demands for oxygen supply, more than 30 times that of resting conditions. Although greater efficiency of these cells to transport oxygen is advantageous to the animal, it could potentially expose the RBC to higher than average risk of oxidant damage. The aim of the present study was to investigate the capacity of RBCs of Pteropus poliocephalus, the greyheaded flying fox, to defend itself against oxidant stress. RBC of flying fox and human were challenged with 2,2′-azobis(2-amidinopropane) dihydrochloride (AAPH). The RBC from the two species were found to have similar reduced glutathione (GSH) reduction kinetics and succumbed to haemolysis at similar rates. However, haemoglobin oxidation was much faster in the flying fox suggesting that the RBC of flying fox may not enjoy as good protection as those of humans in the event of an oxidant stress.     

Comparative nuclear magnetic resonance studies of diffusional water permeability of red blood cells from different species. IX. Australian feral chicken and domestic chicken (Gallus domesticus)

The diffusional water permeability (P _d) of Australian feral chicken and Australian and European domestic chicken red blood cells (RBCs) was measured by a doping nuclear magnetic resonance (NMR) technique. The values of P _d were around 1.7 × 10^–3 cm/s at 15°C, 2.0 × 10^–3 cm/s at 20°C, 2.5 × 10^–3 cm/s at 25°C, 3.7 × 10^–3 cm/s at 30°C, 4.3 × 10^–3 cm/s at 37°C, and 6.1 × 10^–3 cm/s at 42°C, with no significant differences between the three strains of chicken. There was no effect of p-chloromercuribenzene sulphonate on water diffusion. The activation energy of water diffusion was around 37 kJ/mol for all strains of chicken. These results suggest that no changes in the RBC water permeability are correlated with marked alterations in the habitat of chicken introduced to Australia (and that membrane proteins play little role in the diffusion of water across chicken RBC membrane).     

红细胞深低温保存方法的比较研究

探寻可替代甘油并简化复温后洗涤过程的较理想的红细胞低温保护剂,为低温保存红细胞临床应用提供便利。依据血红蛋白溶液浓度与其在波长为412nm处吸光度的关系曲线,建立红细胞回收率评估方法,研究比较了使用5种低温保护剂实现人类红细胞液氮深低温保存效果的优劣。结果表明:35.5%(w/v)甘油的保存效果最佳,其回收率为97.84%;其次为25%(w/v)右旋糖酐40,其回收率为85.08%;再次之为1M海藻糖(加载并孵育17.5h),其回收率为55.35%;回收率最低的两组为1M海藻糖(未经孵育)和10%(w/v)的二甲亚砜,对应的回收率均不足35%。初步推断:右旋糖酐40是一种可望替代甘油的红细胞低温保护剂。     

Comparative nuclear magnetic resonance studies of diffusional water permeability of red blood cells from different species. VIII. Adult and fetal guinea pig (Cavia procellus)

The diffusional water permeability (P _d) of adult, pregnant female and fetal guinea-pig red blood cells (RBCs) was measured by a doping nuclear magnetic resonance (NMR) technique on control cells and following inhibition withp-chloromercuribenzene sulphonate (PCMBS). The values ofP _d were around 5.0 × 10^–3 cm/s at 15 °C, 5.3 × 10-3 cm/s at 20 °C, 6.6 × 10^–3 cm/s at 25 °C, 7.5 × 10^–3 cm/s at 30 °C and 8.6 × 10^–3 cm/s at 37 °C with no significant differences between adult, pregnant female and fetal RBCs. Systematic studies on the effects of PCMBS on water diffusion indicated that the maximal inhibition was reached in 10 min at 37 °C with 0.1 mm PCMBS. The values of maximal inhibition ranged from 70%–77% at 15–30 °C to 57%–63% at 37 °C in the case of adult and from 64%–67% at 15–30 °C to 51% at 37 °C in the case of fetal RBCs. The basal permeability to water was estimated at 1.1 × 10^–3 cm/s at 15 °C ,1.3 × 10^–3 cm/s at 20 °C, 1.6 × 10^–3 cm/s at 25 °C, 2.2 × 10^–3 cm/s at 30 °C and 3.2 × 10^–3 cm/s at 37 °C for adult and slightly higher values for fetal guinea pig RBCs as 1.6 × 10^–3 cm/s at 15 °C, 2.0 × 10^–3 cm/s at 20 °C, 2.4 × 10^–3 cm/s at 25 °C, 2.6 × 10^–3 cm/s at 30 °C and 4.2 × 10^–3 cm/s at 37 °C. The activation energy of water diffusion was around 22 kJ/ mol, with no significant differences between the adult pregnant female and fetal RBCs, and increased to about 40 kJ/mol in the case of adult and pregnant RBCs and 34 kJ/mol for fetal RBCs after incubation with PCMBS in conditions of maximal inhibition of water diffusion. The membrane polypeptide electrophoretic pattern of adult and fetal guinea-pig RBCs was compared with its human counterpart. The guinea-pig membrane contained higher amounts of spectrin (band 1 and 2), whereas the proteins in bands 4.1, 4.2 and 6 were present in lower amounts. Considerable differences in polypeptides migrating in the region of bands 7 and 8 and in front of them were apparent between the two sources of RBC membranes where some bands were present only in the guinea-pig RBC membranes. The adult guinea-pig membranes contained smaller amounts of proteins migrating in band 4.5 and lacked band 8.     

Mathematical Model of Time Needed for the Immune System to Detect and Kill Cancer Cells in Blood

Disfunction of the immune system has generally been blamed for the development of cancers however, the immune system needs time to detect and kill cancer cells. To date, the chance of random collision between immune cells and cancer cells in blood has drawn less attention. In this study we used a random principle to analyse the possibility of collision between immune and cancer cells in blood. With the criterion of p>0.95, the results show that an immune cell needs, for example, five random collisions in order to hit a cancer cell when there are one cancer cell, one immune cell and one normal cell in our consideration; and 14 999 999 random collisions in order to hit a cancer cell when there are one cancer cell, one immune cell and 4 999 999 normal cells in consideration. Furthermore, we analyse how different proliferation rates of cancer cells affect the random collisions between the detecting/killing immune cells and cancer cells.     

Comparative NMR studies of diffusional water permeability of erythrocytes from some animals introduced to Australia: Rat rabbit and sheep

The diffusional water permeability (P _d) of the red blood cell (RBC) membrane of rat, rabbit and sheep, representing some animal species introduced to Australia, has been monitored, by a Mn²⁺-doping ¹H nuclear magnetic resonance (NR) technique using a high-field spectometer operating at 400 MHz. In order to make comparisons with previous measurements on the same species (performed at 25 MHz) an analysis of the influence of instrumental parameters and of the state of blood oxygenation was performed on samples of rat and rabbit blood. It was found that by using a short interpulse delay (around 100 µs) in the Carr-Purcell-Meiboom-Gill sequence, and by performing the measurements of the transverse relaxation time of the water in the cell interior (T _2i) on packed cells (haematocrit >95%), the values of the water exchange time (T _e) obtained with a high-field spectrometer are comparable with those obtained by using a low-field NMR spectrometer. The values of the diffusional water permeability (P _d), calculated from the T _e values, were, at 24.6°C around 10 × 10^–3 cm/s in case of rat and rabbit RBC and around 5 × 10^–3 cm/s for sheep RBC; at 37°C the P _d values were around 16 × 10^–3 cm/s for rat RBC, 14 × 10^–3 cm/s for rabbit RBC and 7 × 10^–3 cm/s for sheep RBC. These values are very close to the P _d values previously reported for the European counterparts of these species. There were no significant differences in the P _d values between laboratory rabbits and wild rabbits, or between castrated males, pregnant or nonpregnant females. These results suggest that no changes in the RBC water permeability are correlated with marked alterations in the habitat of animal species introduced to Australia or by sex hormone status. If the NMR instrument-parameters are carefully chosen then comparative studies of RBC water permeability in different laboratories, at separate locations and using different instruments are meaningful.     

Effects of Duration of Storage and Storage Temperature on Cell Counts of Bovine Blood Samples as Determined by an Automated Haematology Analyser

Analysis of blood cells is an important part of many scientific investigations in the field of cattle herd health. Over the last 30 years, automated blood analysis has all but replaced manual counting of blood cells using counting chambers. The present study investigated the effects of prolonged storage and storage temperatures on cell counts as determined by a haematology analyser. Blood samples from 20 clinically healthy cows were repeatedly analysed with a Cell-Dyn 3500 (Abbott Diagnostika, Delkenheim), within 24 hours after collection and after storage at either 4° C or 20° C. The counts of most blood cells were more stable in samples stored at 20° C than those stored at 4° C. For at least 8 h, the counts of all analysed cell types, with the exception of lymphocytes, remained within ±3 standard deviations that were calculated for fresh samples, provided that the blood was stored at 20° C. Correspondence and offprint requests to: Dr Ulrich Bleul, Klinik für Fortpflanzungskunde, Universit?t Zürich, Winterthurerstrasse 260, 8057 Zurich, Switzerland.     

关于“人红细胞反定型试剂技术审查基本要求”的思考

在“人红细胞反定型试剂”技术审查中,有些红细胞的基本评价指标对于明确试剂质量十分重要,但该产品各申报厂家的评价指标和方法均不同,难以对产品进行充分的评价.因此,本文对于该产品申报资料的基本要求,包括综述资料、主要原材料、主要生产工艺及反应体系、分析性能评估、阳性判断值、稳定性、技术要求、说明书等资料进行了详细的分析,并对临床试验的设计和要求进行了阐述.总之,本文旨在给予申报企业一定的技术指导,使产品能够最大程度地满足临床应用的要求.     

失血性休克过程中红细胞携氧-释氧动力学研究

失血性休克过程中,红细胞自身代谢与功能的变化会导致红细胞携氧-释氧能力的改变。本文对不同休克程度红细胞携氧-释氧能力的变化进行了监测,探讨失血性休克的危重程度与红细胞携氧-释氧能力之间的相互关系,并进行生理生化指标分析。本文采用健康SD大鼠20只(雌雄各10只),建立失血性休克模型,分别在初始、失血20%、30%和40%时采集红细胞,并利用血氧分析仪测定红细胞携氧-释氧热力学参数P50,效应时间Tc50、Tr50,同时检测了2,3-DPG、pH值、葡萄糖和乳酸脱氢酶的变化。实验结果表明:随着休克程度的加深,P50值不断增大,乳酸脱氢酶增加,而Tr50、pH值、葡萄糖均呈现减小的趋势,Tc50变化不明显;红细胞携氧能力相对稳定,而向组织释氧的能力仅略有增加,机体氧耗无法得到满足导致多器官衰竭。     

A Comparative Study of Erythrocytic Total, Free, Short-chain Acyl and Long-chain Acyl Carnitine Concentrations in Arabian Camel ( Camelus dromedarius ) with Different Species

Carnitine plays a critical role in lipid metabolism. Carnitine deficiency may adversely affect the oxidation of fatty acids and further aggravate abnormal lipid metabolism. Its presence is considerable in tissues that utilise fatty acids as an important source of energy, such as the heart and skeletal muscle. The presence of total (TC), free (FC), short-chain acyl (SC) and long-chain acyl (LC) carnitine was shown for the first time in one-humped Arabian camel (Camelus dromedarius) erythrocytes. The results showed that concentrations of FC, AC, SC and TC in camel erythrocytes were significantly higher when compared with bovine, rabbit, rat and humans; however, these carnitine fractions were not significantly different when compared with sheep. Moreover, the concentration of LC in camel erythrocytes was significantly higher when compared with rabbit, rat and humans, but there was no significant difference compared with either sheep or bovine. The results showed that there were significant variations in the ratio of acyl carnitine (AC) to FC among the species erythrocytes studied. The ratio in camel erythrocytes was significantly higher when compared with rat, but there was no significant difference in this ratio in camel erythrocytes when compared with bovine, rabbit, sheep and humans. The higher carnitine concentrations and a higher proportion of AC in erythrocytes of the Arabian camel suggest an adaptive mechanism that could be common to desert animal species.     

Normal Haematopoiesis, Cellular Components and M/E Ratio in the Bone Marrow of the Black-headed Gull (Larus ridibundus)

In order to study the morphology of haematopoietic cells of the black-head gull, bone marrow samples were collected from the proximal tibiotarsus bone of 50 clinically healthy adult gulls. The bone marrow smears were stained using the Giemsa technique. The results indicated that the development and formation of blood cells in the bone marrow of gulls were similar to other birds, whereas the morphology of the cells was similar to chickens, ducks and quails. The mean myeloid/erythroid (M/E) ratio was 1.23 ± 0.17, the mean erythroid percentage was 39.91 ± 3.26%, the mean myeloid percentage was 49.37 ± 4.86%, the mean thrombocyte percentage was 5.95 ± 0.79% and the mean percentage of all other cells percentage was 4.77 ± 0.53%. Correspondence and offprint requests to: M. Tadjalli, Department of Basic Sciences, School of Veterinary Medicine, Shiraz University, Shiraz 71345, Iran     

新生婴儿与老年人红细胞物理特性对比实验

目的 通过比较新生婴儿与80岁以上老年人红细胞物理特性(细胞表面积、体积和剪切弹性模量)及与总胆固醇、糖化血红蛋白等生理生化参数的相关关系,分析不同年龄和生化参数对红细胞物理特性的影响.方法 采用微量吸管技术测量新生婴儿和80岁以上老年人的红细胞表面积、体积和剪切弹性模量,对数据进行统计分布分析、相关性分析和回归分析....     

A Study of the Distribution of Total, Free, Peptide-bound and Protein-bound Hydroxyproline in the Plasma of Arabian Camel ( Camelus dromedarius )

This study was conducted to determine the presence of total, free, protein bound and peptide bound hydroxyproline (Hyp) in the plasma of the Arabian camel (Camelus dromedarius) as compared to the sheep. Results showed that the Hyp pool of sheep plasma was composed of free, protein bound and peptide bound hydroxyproline. On the other hand, only free and protein bound Hyp contributed to the Hyp pool of the camel plasma; peptide bound Hyp was not detected. Camel plasma showed a significantly lower content of total Hyp (p<0.001), however, a higher level (p<0.001) of protein bound Hyp as compared to sheep. The observed variations in Hyp distribution may result from differences in the Hyp derivatives and/or compositions in these species. This is the first report describing the distribution of Hyp in camel plasma.     

有核红细胞采用XN1000血液分析仪计数与人工计数结果比较

目的 Sysmex XN1000b型全自动血细胞分析仪检测的有核红细胞结果的有效性和准确性,通过金标法人工涂片检测进行验证。方法 2017年1月~2017年3月本院经过Sysmex XN1000b型全自动血细胞分析仪检测有核红细胞结果阳性标本213例进行仪器结果与人工涂片结果进行准确性验证,对随机抽取100例有核红细胞结果为零的标本验证阴性符合率验证。结果 采用Sysmex XN1000b型全自动血细胞分析仪计数有核红细胞,真阳性率100.00%,假阳性率2.09%,假阴性率0（＜5%）,真阴性率96.70%,阳性结果符合率97.56%。结论 Sysmex XN1000b型全自动血细胞分析仪检测的有核红细胞计数准确性高,阴阳性符合率好,结果可靠,有核红细胞计数可以作为临床报告项目。