活动力不足精子的线粒体变化—形态学和计量学研究

通过电镜观察和体视学方法，研究活动力不足精子的线粒体变化。结果是运动力不足的精子尾部中段存在较多胞质，线粒体排列方式呈现异常，线粒体内部的结构有退化现象。运动力不足的子线粒体的体密度、外膜面密度和面数密度均小于对照组，线粒体的外膜比表面，则大于对照组，而线粒体的平均体积及平均外表面积和对照组比无差异。     

Pain relief by somatostatin in attacks of cluster headache

The pain relieving effect of somatostatin treatment during 72 attacks of cluster headache in 8 male patients was compared to treatment with ergotamine or placebo in a double-blind study. Infusion of somatostatin (25 micrograms/min for 20 min i.v.) reduced the maximal pain intensity and the duration of pain significantly compared to placebo treatment, and to a degree comparable to ergotamine tartrate treatment (250 micrograms i.m.). The results obtained provide new information concerning the possible mechanism of cluster headache attacks and suggest a new therapeutic approach.     

A fluorometric assay for measurement of protoporphyrinogen oxidase activity in mammalian tissue

The oxidation of protoporphyrinogen to protoporphyrin, which is catalyzed by the enzyme protoporphyrinogen oxidase, is an important step in heme biosynthesis. We describe a fluorometric assay for protoporphyrinogen oxidase activity in mammalian tissues which measures the conversion of the non-fluorescent protoporphyrinogen to the fluorescent protoporphyrin. Using these assay conditions, the mean level of protoporphyrinogen oxidase activity in sonicated rat liver mitochondria was 12.3, and in sonicated human cultured skin fibroblasts was 1.97 nmol protoporphyrin/mg protein/h.     

Recombinant human erythropoietin: effects on frataxin expression in vitro

BACKGROUND: Friedreich's ataxia (FRDA) is a neurodegenerative disorder caused by decreased expression of the protein frataxin, recently described to be an iron chaperone for the assembly of iron-sulphur clusters in the mitochondria, causing iron accumulation in mitochondria, oxidative stress and cell damage. Searching for compounds that could possibly influence frataxin expression, we found that the cytokine recombinant human erythropoietin (rhuEPO) significantly increases frataxin expression by a still unknown mechanism. MATERIALS AND METHODS: Isolated lymphocytes from FRDA patients, isolated human cardiac cells (fibroblasts and myocytes) from patients undergoing heart transplantation and P19 mouse cells (neuronal typ), were incubated with different concentrations of rhuEPO, and immunoblot was carried out for the detection of frataxin. RESULTS: We show for the first time that the cytokine recombinant human erythropoietin (rhuEPO) can, additionally to its reported neuro- and cardioprotective properties, increase frataxin expression in vitro. We show that rhuEPO significantly increases frataxin expression in primary lymphocytes from patients with Friedreich's ataxia. Further we show that rhuEPO can also increase frataxin expression in many other cell types; among them the most affected cell types in FRDA such as neurones and cardiac cells. CONCLUSIONS: Our results provide a scientific basis for further studies examining the effectiveness of this agent for the treatment of FRDA patients.     

Memory for pain.

Memory for head pain was assessed by means of the McGill Pain Questionnaire (MPQ). Sixteen neurosurgical patients were divided into two groups in order to examine the decay of memory over time; one group recalled pain after 5 days and the other recalled pain after one day and then again, after 5 days. Contrary to expectations, the recall of pain was surprisingly accurate. The memory for pain showed littled decay over time. The small subgroup of patients who made specific errors when recalling their pain comprised women who had high levels of pain and affect at the initial assessment. Overall, the findings provide some welcome reassurance about the accuracy and reliability of pain reports from memory.     

Assessment of protease-binding by alpha 2 macroglobulin in multiple sclerosis

Following reports that α₂-macroglobulin (α₂M) is abnormal in multiple sclerosis (MS), we assessed the interaction of α₂M with a model protease bovine cationic trypsin. Two parameters were investigated: (1) the binding of α₂M to trypsin, and (2) the ability of the trypsin-α₂M complexes to hydrolyze benzoyl arginine ethyl ester. In both respects, MS α₂M was found to be similar to control α₂M.     

Dynamics of the binding capacity of plasma sex hormone binding globulin (SHBG) for testosterone and dihydrotestosterone during puberty

Plasma sex hormone binding globulin binding capacity (SHBG-b.c.) has been evaluated in 203 normal subjects (114 males and 89 females) aged 3 to 51 years. The subjects were divided into groups: prepubertal, early pubertal (Tanner's Stages 1 and 2), late pubertal (Tanner's Stages 4 and 5) and adult.In both sexes, plasma mean values of SHBG binding capacity both for dihydrotestosterone (DHT) and testosterone (T) were significantly higher in prepubertal subjects, falling during puberty to adult levels.During pubertal development DHT-BG binding capacity and T-BG binding capacity showed different plasma values with respect to sex and phase of puberty.Our data do not support an absolute relationship between sex hormones and SHBG binding capacity, but suggest other mechanisms as well: (a) SHBG modifies its physicochemical properties during puberty, or (b) the binding capacity is the result of a pool of proteins which modifies its composition during pubertal evolution.     

Analytical subcellular fractionation studies on rat liver and on isolated jejunal enterocytes with special reference to the separation of lysosomes, peroxisomes and mitochondria.

1. Enterocytes were isolated from rat jejunum and characterized morphologically. 2. Attempts to separate the enterocyte subcellular organelles, characterized by their marker enzymes, with isopycnic centrifugation were unsuccessful but good separation of peroxisomes, lysosomes and mitochondria was achieved by sedimentation through a shallow sucrose density gradient with a super-imposed inverse gradient of low-molecular-weight dextran. 3. The properties and enzyme activities of the principal subcellular organelles in rat liver cells and enterocytes were compared.     

Acute pain in an emergency clinic: Latency of onset and descriptor patterns related to different injuries

Features of acute pain were examined in patients at an emergency clinic. Patients who had severe, life-threatening injuries or who were agitated, drunk, or ‘in shock’ were excluded from the study. Of 138 patients who were alert, rational and coherent, 51 (37%) stated that they did not feel pain at the time of injury. The majority of these patients reported onset of pain within an hour of injury, although the delays were as long as 9 h or more in some patients. The predominant emotions of the patients were embarrassment at appearing careless or worry about loss of wages. None expressed any pleasure or indicated any prospect of gain as a result of the injury.The occurrence of delays in pain onset was related to the nature of the injury. Of 46 patients whose injuries were limited to skin (lacerations, cuts, abrasions, burns), 53% had a pain-free period. Of 86 patients with deep-tissue injuries (fractures, sprains, bruises, amputation of a finger, stabs and crushes), only 28% had a pain-free period. The McGill Pain Questionnaire was administered to patients who felt pain immediately after injury or after a delay, and revealed a normal distribution of sensory scores but very low affective scores compared to patients with chronic pain. The results indicate that the relationship between injury and pain is highly variable and complex.     

Drug utilization patterns in chronic pain patients

In the population of chronic pain patients seen at multidisciplinary pain clinics, excessive and/or inappropriate medication use is a frequent problem. This study examined differences between chronic pain patients who used no addicting medication (30% of the sample of 131 patients), those who used narcotic but not sedative medications (33%) and those who used both narcotic and sedative medications (37%). Patients in the narcotic and narcotic-sedative groups had undergone significantly more pain-related hospitalizations and surgeries than those in the no addicting drugs group. Narcotic-sedative patients spent significantly more money on pain medication per month, reported significantly greater physical impairment, and had higher MMPI hypochondriasis and hysteria scores when compared to the other patients. The findings are interpreted in light of the hypothesis that certain patients show greater readiness to complain of and seek help for physical symptoms.     

A pilot study of the treatment of outpatients with chronic pain: symptom control, stimulus control and social system intervention.

Of 6 outpatients with chronic pain, 5 completed therapy based on a 3-part treatment package designed to provide symptom control, stimulus control and social system modification. Each of the components of the treatment package resulted in therapeutic change. A mean of 35.8 weekly hour long therapy sessions resulted in statistically significant decreases in pain, hopelessness, depression and analgesic medication intake. Generally, these improvements were maintained at 6 months and 1 year follow-up. This study is consistent with the notion that chronic pain is maintained by a combination of inter- and intrapersonal factors. A controlled comparison of this treatment program with other treatments for chronic pain is indicated.     

Comparative study between the Ves-matic and microerythrocyte sedimentation rate method

In this study, Ves-matic erythrocyte sedimentation rate and micro-erythrocyte sedimentation rate methods were compared on 96 subjects mean age 4.9+/-4.3 years. Ves-matic erythrocyte sedimentation rate an automated method, and micro-erythrocyte sedimentation rate method required minimal bloods are various approaches on the erythrocyte sedimentation rate. An important relationship between that the Ves-matic and micro-erythrocyte sedimentation rate was revealed. Although there was significantly correlation between these two methods, they are different to use interchangeably. Our study implies that "Micro-Automated Erythrocyte Sedimentation Rate Systems" could be developed which require a few amounts of blood and study automatically.     

Binding of 3,5-diiodotyrosine to serum proteins.

The reversible binding of 3,5-diiodotyrosine (DIT) to human and bovine serum protein and to purified human serum prealbumin and human and bovine albumin has been studied by equilibrium dialysis. Maximum binding occurred at pH 8.6-9.0. Human serum bound DIT less than did bovine serum. Adult ox and fetal calf sera showed similar binding. The main DIT-binding protein of human serum was prealbumin. It showed a single affinity site with a Ka of 0.85 X 10(6) M-1 at pH 8.6 and 0.40 X 10(6) M-1 at pH 7.4. The affinity constant of serum albumin for DIT was 2.8 X 10(3) M-1 at pH 8.6. The elevated binding of DIT to bovine serum is essentially due to albumin whose affinity constant for DIT is 16-times higher than that of human serum albumin. Fetuin was not responsible for any noticeable DIT binding in fetal calf serum.     

A novel specific assay of 25-hydroxy vitamin D.

The synthesis of 25-hydroxy-[26-2H3]vitamin D3 is described. A fixed amount of this compound (usually 250 ng) is added to a fixed amount of serum (usually 2.5 ml) and the mixture is extracted with a chloroform/methanol mixture. The extract is chromatographed on a Sephadex LH-20 column together with a trace amount of 25-hydroxy-[263H3]vitamin D3. The chromatographic fraction corresponding to 25-hydroxy vitamin D3 is converted into trimethylsilyl ether and the amount of unlabeled 25hydroxy vitamin D3 is determined from the ratio between the mass fragmentographic recording at m/e 131 (base peak of unlabeled 25-hydroxy vitamin D3) and m/e 134 (base peak 25-hydroxy-[26-2H3]vitamin D3). The relative standard deviation of the method was about 5%.     

Comparison of Inverted Centrifugation, Saline Washing, and Dextran Sedimentation in the Preparation of Leukocyte-Poor Red Cells

Three methods (inverted centrifugation; inverted centrifugation followed by continuous-flow washing in 5 per cent dextrose-saline; and dextran sedimentation followed by continuous-flow washing in 5 per cent dextrose-saline) were evaluated for the preparation of leukocyte-poor red blood cells. Residual leukocyte contents were 0.32, 0.40, and 0.16 billion per 100 g of erythrocytes and erythrocyte losses were 24, 42, and 35 per cent, respectively, for the three methods. The mean removal of leukocytes per unit after inverted centrifugation was 80 per cent. Continuous-flow washing after inverted centrifugation did not remove additional leukocytes. Dextran sedimentation removed a mean 92 per cent of leukocytes. Residual dextran was not detected in the terminal wash solution. These findings suggest dextran sedimentation may be useful for preparing leukocyte-poor red blood cells for preventing sensitization to leukocytes in high risk recipients such as transplantation candidates.     

Application of nicotinamide-adenine dinucleotide analogs for clinical enzymology: a spectrophotometric method for clinical analysis of lactate dehydrogenase patterns with the use of a nicotinamide-adenine dinucleotide analog.

The activities of porcine lactate dehydrogenase (LHD) isoenzymes were analyzed using nicotinamide-adenine dinucleotide (NAD) or its analogs as a cofactor, with varying concentrations of L-lactate from 13 to 530 mM. The greatest differences between H4-type and M4-type isoenzymes in reaction rates were observed when their activities were compared in a reaction mixture containing 530 mM lactate and NAD, and also in a system of 13 mM lactate with thionicotinamide-hypoxanthine dinucleotide as a cofactor. The ratio of the LDH activity exerted in the former reaction mixture to that exerted in the latter was termed the N/T value. The N/T values of porcine H4 and M4 isoenzymes were 0.49 and 9.33, respectively. The N/T values of other three isoenzymes (H3M1, H2M2 and H1M3) were calculated by assuming that the single subunits H1 and M1 contribute one-fourth of the values of 0.49 and 9.33, respectively, and a given isoenzyme which is a combination of four subunits of H and M comprises the sum of their values. The calculated values agreed fairly well with the experimental results. The N/T value method was found to be applicable to human LDH isoenzymes, and sera from various patients were analyzed in comparison with hormonal sera. The method is particularly suitable for the numerical expression of LDH isoenzyme profiles.     

Influence of energy-coupling inhibitors on the respiration of tightly-coupled human skeletal muscle mitochondria

Abstract A standardized preparation method for the study by polarographic technique of human skeletal muscle mitochondria is described. The mitochondria thus obtained are tightly coupled, exhibiting high respiratory control ratios with classical responses to energy-coupling inhibitors; they are also able to accumulate calcium, this uptake being blocked by inhibition of the phosphate carrier system. Arsenate uncoupling and integrity of the electron transport chain are demonstrated polarographically. The properties of human skeletal muscle mitochondria are generally similar to those of rat-skeletal muscle and-liver mitochondria; differences are discussed.     

Bench to Bedside: The Role of Mitochondrial Medicine in the Pathogenesis and Treatment of Cellular Injury

Interest in the study of mitochondria has undergone a revival. The term mitochondrial medicine developed around evidence pointing to the mitochondria as a logical target for therapy. This article reviews the normal functions of mitochondria and integration of mitochondrial processes in cells. Changes in mitochondria that occur in ischemia and reperfusion, production of reactive oxygen species by mitochondria, stimulation of apoptosis, and roles of mitochondria in sepsis also are reviewed. The authors also review therapies that are based on targeting drugs to mitochondria, regulating calcium availability, substrate preferences, and activation of poly (ADP-ribose) polymerase and apoptosis. The purpose of this article is to provide a framework for emergency medicine academicians to understand the roles of mitochondria in pathology and to facilitate the transition of this information into therapeutic strategies based on mitochondrial medicine. For individuals interested in the biochemistry of mitochondria, knowledge of this fundamental organelle is expanding at a rapid rate.     

A simple method for preparing neocyte-enriched leukocyte-poor blood for transfusion-dependent patients

Recent treatment for patients with thalassemia and chronic anemia involves transfusion of young red cells (YRBCs) or "neocytes." We developed a technique enabling YRBCs to be separated based on their buoyant density in autologous plasma during centrifugation. Following this procedure, measurement of pyruvate kinase (PK), an age-dependent red cell enzyme, showed neocyte enrichment in the top one-third of the RBC layer corresponding to a mean of 47.5 percent of the total PK present in the unfractionated unit. To provide a neocyte transfusion preparation with an acceptable hemoglobin content, the top one-third fraction from each of three bags of blood was pooled. Leukocytes were removed from this "neocyte unit" by an initial sedimentation with 6 percent hydroxyethyl starch (HES) followed by filtration through a filter (IG 500, Imugard). This process resulted in removal of 99.3 +/- 0.5 percent (mean +/- SD) of the leukocytes with a mean RBC recovery of 89.5 +/- 5.5 percent and a final hemoglobin content of 53.4 +/- 2.3 g. Tests for plasma-free hemoglobin and HES in the supernatant of the final transfusion product gave acceptable mean values of 28 mg per dl and 3.0 mg per ml. Autologous mean RBC survival of Cr51-labeled YRBC fractions was 41.8 +/- 2.9 days (n = 5). This technique yields neocyte enrichment superior to that achieved using a cell processor (model 2991, IBM) and has the added advantage of being less costly to prepare ($45.00 [1984] U.S. per YRBC unit as compared to an estimated $135.00 [1984] U.S., IBM) and more economical in terms of blood use.     

The systemic administration of local anaesthetics produces a selective depression of C-afferent fibre evoked activity in the spinal cord

An electrophysiological analysis of the antinociceptive effects of systemic lidocaine and its longer acting primary amine congener, tocainide, has been performed in the decerebrate-spinal unanaesthetised rat. Neither of these local anaesthetic drugs when administered systemically in doses of up to 10 mg/kg (lidocaine) or 100 mg/kg (tocainide), produced any evidence of a block in the conduction of action potentials in A beta, A delta or C primary afferents. The local anaesthetics also failed to reduce mustard oil induced neurogenic extravasation, a test of cutaneous C-fibre terminal function. Lidocaine produced a transient (1-2 min) depression in monosynaptic reflexes at doses of greater than or equal to 1 mg/kg while tocainide had no effect on this reflex at any dose up to a 100 mg/kg. Both drugs, however, significantly suppressed the C-fibre evoked polysynaptic reflex generated by stimulating the sural nerve. The tocainide effect was longer lasting with less action on the short latency A beta-evoked reflex than lidocaine. The reflex activity in hamstring flexor alpha-motoneurones evoked by pinching the toes of the ipsilateral hind paw was reduced by both drugs but not abolished. Thermal and noxious chemical evoked reflexes were, however, completely suppressed by the local anaesthetic drugs, again with a longer action from tocainide. These results demonstrate that the systemic administration of drugs which increase the inactivation of sodium channels can produce a selective central block of certain types of afferent evoked activity in the spinal cord. There are resemblances between the selective C-fibre suppressing actions of systemically administered local anaesthetic and the pharmacological actions of narcotic opiates which may represent a similar mechanism for the analgesic action of these quite different classes of drugs.