Transferase contents in lymph and blood in fever reaction]

The aim of the present investigation was the study of the content of alanine, asparagine-transaminases, gamma-glutamyl transferase and their isoenzymes, as well as leucine aminotransferase in the lymph of thoracic duct, hepatic and intestinal lymph and peripheral blood in dynamics of fever reaction of various duration in the experiments on rabbits. Irrespective of its duration, the fever was followed by a significant activation of the enzymes in the body fluids. However, in many-day fever reaction, a rise of enzymes level in the lymph was more prolonged than that in the blood. The above studies make it possible to assume that the released enzymes in fever reaction are primarily resorbed by lymphatic capillaries and their activity indices in the blood serum are largely evidenced by the transport function of the lymphatic system.     

Phosphatase activity in the lymph during a fever reaction]

Activity of acid and alkaline phosphatases, as well as isoenzymes of alkaline phosphatase in the lymph of the thoracic lymphatic duct, hepatic lymph and the peripheral blood have been studied on rabbits in the dynamics of the fever reaction of different duration. The fever reaction was followed by enzyme activity increase in all the body biologic fluids. However the degree of increase of their activity in the lymph was greater that that in the blood. Our data indicate that in the transport of phosphatases released from the tissues in the common circulation the essential role is played by the lymphatic system, the resorption and transport functions of which significantly characterise the dynamics and the level of their changes in the blood in fever reaction.     

Cytological composition of lymph in fever reaction

This work deals with an investigation into the lymph cytologic composition of thoracic lymph duct of rabbits in fever reaction (FR) of various duration. FR was accompanied by quantitative and qualitative shifts in lymph cytologic composition. There was an alternative rise and fall of the leucocyte number in the first hours of fever. The number of little and medium leucocytes decreased while the number of eosinophiles, insufficiently differentiated cells-blasts, large lymphocytes, prolymphocytes increased. Our investigations revealed a significant role played by the lymphatic system in lymphoid cells mobilization in FR, which is evident by a considerable lymphocyte number gaining entrance to the blood through thoracic duct.     

Morpho-functional changes in the lymph nodes during fever reaction]

Lymph nodes (mesenteric, popliteal, cervical) of rabbits in fever reaction of different duration have been studied in our work. As a whole morpho-functional changes in lymph nodes in fever reaction indicate the increase of their functional activity: hyperplasia of lymphatic substance with the growth of lymphocytes number and slightly differentiated lymphoid cells in follicles and paracortical zone, hyperplasia of pulposus bands, the signs of macrophagal reaction and plasmatization of lymph nodes are to be observed and all these create prerequisites for the increase of tensity of cellular and humoral immunity. Simultaneously the signs of destruction of cellular elements--lymphocytolysis in the porta tract and the growth of number of PAS-positive cells in the parenchyma of the nodes take place.     

Effect of the fever reaction on the kallikrein-kinin system of the lymph and blood

The dynamics of kallikrein-kinin system components in the lymph of thoracic duct and blood following fever reaction of various duration has been studied in the experiment on rabbits. The experiments have shown that in prolonged fever reaction there are qualitative disorders in kinin system components ratio indicative of break in the system of regulatory links, its exhaustion and transformation of the physiological reactions into the pathogenic ones. One may assume that changes in kallikrein-kinin system activity in body fluid are the links in the complex of pathogenic disorders in the organ and system functional activity during prolonged fever reaction.     

Enzyme activities in thoracic duct lymph and plasma of anaesthetized, conscious resting and exercising dogs

The significance of changes in lymph flow for the extracellular distribution and transport of cellular enzymes and for the level of enzyme activities in plasma was investigated. Specimens of thoracic duct lymph were obtained from an extracorporal lymph shunt in anaesthetized, conscious resting and treadmill exercising dogs (6 km X h-1 for 1 h) The activity of 10 enzymes and of protein content in lymph and plasma were studied, as well as lymph flow, lymphatic transport, and the lymph-plasma ratio of these compounds. Lactate, pH, and blood gases were monitored in venous blood. Lymph flow of 0.80 ml X min-1 in anaesthetized dogs more than doubled (to 1.86 ml X min-1) when the animals were conscious and resting. In anaesthetized dogs lymph enzyme activity was higher only for enzymes of predominately hepatic origin, such as choline esterase (CHE) and alanine aminoferase (ALAT), and was lower for aspartate aminotransferase (ASAT) and aldolase (ALD). In conscious dogs, due to activation of the skeletal muscle "tissue pump", lymphatic transport of enzymes with rather high activity in skeletal muscle, and of protein, is significantly enhanced. Enzyme activities in plasma, however, did not differ between the groups. Lymph-plasma activity ratios higher than one were found for lactate dehydrogenase (LDH), malate dehydrogenase (MDH), ASAT, creatine kinase (CK), ALD, and phosphohexose isomerase (PHI). Exercise stimulated lymph flow up to 4.9 ml X min-1, and increased the lymphatic activities of those enzymes with a lymph-plasma ratio higher than unity, these enzymes increasing in the plasma due to the highly increased lymphatic transport.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of isothiuronium alkylamidines and adeturon on the activity of lactate dehydrogenase and its isoenzymes in rat blood serum and organs

A study was made of the effect of four types of S-omega-carboxamidinoalkyl isothiourea, differing in the carbon chain length between amidine and isothiuronium groups, and adeturon on activity of lactate dehydrogenase (LDG) and its isoenzymes in blood serum and organs of rats. Adeturon and compounds with one and three methylene groups elicit a radioprotective effect whereas substances with an even number of methylene groups have no such an effect. The obtained data indicate that changes in activity of LDG and its isoenzymes depend upon the structure of the substance applied. This makes LDG an adequate model in comparative studying the specificity of biochemical effects induced by the radioprotective agents and substances similar in their structure but having no radioprotective efficiency.     

Lymphatic transport of cellular enzymes from muscle into the intravascular compartment.

In an experimental study, employing anaesthetized dogs, it was investigated whether cellular enzymes from peripheral skeletal muscle get into the circulating blood by diffusion across capillary membranes or by lymphatic transport. In the experimental group 1, the animals were anaesthetized only. The plasma activities of the four enzymes measured--lactate dehydrogenase, aspartate aminotransferase, alanine aminotransferase, creatine kinase--did not show any mentionable change during a time period of 6 h. In group 2 one hind limb of each animal was moved passively for 1 h. Alanine aminotransferase remained unchanged in plasma, the activities of the three other enzymes increased significantly. In group 3 one hind limb was made hypoxic by clamping the femoral blood vessels for 1 h. No activity changes were observed. When the period of hypoxia was followed by a 1-hour period of passive movement in group 4, the alterations in plasma activities were almost identical to those observed in group 2. In group 5 the experimental procedure was as in group 4, in addition the lymph from the thoracic duct was quantitatively withdrawn. The enzyme activities in plasma revealed a tendency to decrease rather than increase. Lymph flow increased significantly as well as the lymphatic activities of those enzymes which have high intracellular activities in muscle. The results prove, that enzymes from muscle are transported from the interstitial into the intravascular compartment mainly by lymphatic transport. Indications were found that the interruption of blood flow in one hind limb did not result in an enzyme release from muscle cells. It is discussed how changes in lymph flow, occurring during physical exercise for example, affect enzyme activities in plasma.     

Serum enzymatic activity in oncological patients treated with carminomycin]

Karminomycin effect on the activity of some serum enzymes, such as hexokinase (HK), lactate dehydrogenase (LDG), its isoenzymes and glucose-6-phosphatase (G-6-P-ase) was studied. Biochemical assays were applied to 52 patients with neglected malignant tumors. The course dose of the drug was on the average 72mg. The objective antitumor effect was registered in 15 patients. A reliable increase in the values of LDG-5 and G-6-P-ase was observed after the treatment course in the combined group consisting of all the patients subjected to the biochemical assay. Normalization of the serum enzyme spectrum was observed in 15 patients effectively treated with karminomycin: activity of HK and the cathode fractions of LDG decreased. When treatment with karminomycin was ineffective (37 cases), the changes in the enzymatic activity recorded before the treatment further aggraviated. It was found that the level of G-6-P-ase in the patients' treated with karminomycin increased independent of the treatment effect which was probably associated with its toxic effect on the liver. The increase was reversible.     

Use of differences in substrate inhibition for determination of the interrelationship between molar fractions of lactate dehydrogenase subunits]

A kinetic method of estimating the ratio of mole quota of H and M human lactate dehydrogenase (LDG) subunits is proposed, based on changes in substrate inhibition of LDG isoenzymes with lactate. Stability of kinetic constants for a long period of time is demonstrated. The dependency of activities ratio under low and high substrate concentration on the contribution of mole quota of LDG M subunits is studied. The correlation of experimental and theoretical values is shown to be: r=0.998 p less than 0.001. A comparison is carried out of the content of LDG subunits molar quotas in artificial mixtures with electrophoretic experimental data, a good coinsidence of these values being registered. The informative importance of the method described with standard methods of the estimation of LDG isoenzyme systems is discussed. No effect of components of human diploid cells homogenate and an insignificant effect of blood serum components on kinetic constants of LDG isoenzymes is registered. A dependency of variation coefficients on the enzyme activity is studied, minimal omegan value being 0.6%. The applicability of the method described for the calculation of quantitative content of both LDG subunits in natural objects (blood serum, diploid cell homogenate etc.) is demonstrated.     

胃癌及胃壁组织内淋巴管分布的研究

本文首次用５′－Ｎａｓｅ－ＡＬＰ双重组织化学方法观察了２０例胃癌组织和癌旁胃壁组织内淋巴管的细微分布。在光镜下毛细淋巴管和淋巴管５′－Ｎａｓｅ染色强阳性,管壁显示明显的棕色或深棕色,而毛细血管和血管的ＡＬＰ反应显示强阳性,管壁呈明显的蓝色。据此可将淋巴管、血管区别开来、本研究发现胃癌组织内有较多的淋巴管、毛细淋巴管以及较多的棕色实性条状组织,这些条状物可能是新生的毛细淋巴管。上述结果为研究癌组织内淋巴管的分布及胃癌淋巴道转移,提供了可靠的实验方法。     

Transport function of lymph nodes in body antiorthostatic posture

The spontaneous contractile activity of isolated lymph nodes and the lymph flow from intestine lymphatic vessel in antiorthostatic posture of rats with an inclination angle of 30 degrees during 7-14 days, was decreased. Contractions of the rat lymph nodes in response to actions of adrenaline, acetylcholine and histamine were diminished. There are changes of biochemical components of lymph and blood plasma with simultaneous decrease of the blood plasma volume. It is concluded that the lymphatic system on antiorthostatic posture plays the compensatory role with the purpose of stabilization of homeostasis in the brain.     

Impairments in the intrinsic contractility of mesenteric collecting lymphatics in a rat model of metabolic syndrome

Numerous studies on metabolic syndrome (MetSyn), a cluster of metabolic abnormalities, have demonstrated its profound impact on cardiovascular and blood microvascular health; however, the effects of MetSyn on lymphatic function are not well understood. We hypothesized that MetSyn would modulate lymphatic muscle activity and alter muscularized lymphatic function similar to the impairment of blood vessel function associated with MetSyn, particularly given the direct proximity of the lymphatics to the chronically inflamed adipose depots. To test this hypothesis, rats were placed on a high-fructose diet (60%) for 7 wk, and their progression to MetSyn was assessed through serum insulin and triglyceride levels in addition to the expression of metabolic and inflammatory genes in the liver. Mesenteric lymphatic vessels were isolated and subjected to different transmural pressures while lymphatic pumping and contractile parameters were evaluated. Lymphatics from MetSyn rats had significant negative chronotropic effects at all pressures that effectively reduced the intrinsic flow-generating capacity of these vessels by ～50%. Furthermore, lymphatics were remodeled to a significantly smaller diameter in the animals with MetSyn. Wire myograph experiments demonstrated that permeabilized lymphatics from the MetSyn group exhibited a significant decrease in force generation and were less sensitive to Ca(2+), although there were no significant changes in lymphatic muscle cell coverage or morphology. Thus, our data provide the first evidence that MetSyn induces a remodeling of collecting lymphatics, thereby effectively reducing their potential load capabilities and impairing the intrinsic contractility required for proper lymph flow.     

Occurrence of cysteine proteinase inhibitors in cells of the monocytic-histiocytic system. An immunohistochemical investigation

Human tissues are known to contain two low molecular weight (MW about 12,000) cysteine proteinase inhibitors, i.e. an acid inhibitor (ACPI) with pI 4.7-5.0 and a neutral inhibitor (NCPI) with pI 6.0-6.5. ACPI is abundant in cornifying epithelial tissues and in the dendritic reticulum cells of germinal centres of the lymph nodes. NCPI is abundant in lymphatic tissue and is known to be synthesized and released by mononuclear phagocytes. In this report NCPI was localized immunohistochemically in the epitheloid cells of most sarcoidotic lymph nodes, in lymph node macrophages after lymphangiography and in alveolar macrophages, while no ACPI could be demonstrated in the same cells by similar methods. These inhibitors were not demonstrable in lymph node sinus histiocytosis. Peripheral blood monocytes did not exhibit any NCPI immunoreactivity. In occasional blood monocytes anti-ACPI serum gave a weak reaction, the specificity of which is questionable. These data suggest that studies on cysteine proteinase inhibitors reveal basic differences in the various histiomonocytic cells and possibly differences in their functional stages.     

Morphofunctional changes in the hemo- and lymphomicrocirculatory bed of the uterus and its regional lymph nodes in venous congestion during pregnancy

Interrelations of the blood and lymphatic systems of the uterus have been examined in rats at pregnancy complicated with the ++phlebo-occlusive syndrome. Blood stream impediment in the caudal vena cava results in increasing diameter of the arterioles, capillaries and venules of the endo- and myometrium. Certain disturbances of blood circulation in the uterus at the ++phlebo-cclusive syndrome in the pregnancy animals, as a rule, reflect in the uterine lymph outflow; this is demonstrated as dilatation and deformity of the lymphatic vessels and capillaries, appearance of protrusions of the lymphatic vessels wall. There is a definite co-ordination in the reaction to the caudal vena cava occlusion in the intra-, ++extra-organic uterine lymphatic bed and in its regional lymph nodes, manifested as the following morphological signs: decreasing relative volume, that the lymphoid tissue occupies and increasing relative volume of the cortical and medullary intermediate sinuses.     

Immunohistochemical method identifies lymphovascular invasion in a majority of oral squamous cell carcinomas and discriminates between blood and lymphatic vessel invasion.

Tumor invasion into blood and/or lymphatic channels is an important component of cancer staging and prognosis. Standard pathological methods do not provide sufficient contrast to discriminate between invasion into each type of vessel and are complicated by tissue retraction artifacts. We evaluated the ability of a triple-stain immunohistochemical method, combining cytokeratin, CD34, and podoplanin stains in a single section, to distinguish blood from lymphatic vascular invasion in oral squamous cell carcinoma and confirmed its results using multispectral analysis. The triple-stain method was significantly more sensitive in detecting invasive events than the standard hematoxylin and eosin staining method and easily discriminated between blood and lymphatic vessel invasion. Invasive events were present in blood and/or lymphatic vessels in the majority of patients with and without presentation of lymph node metastasis, indicating that vessel invasion in this cancer model is common and is not a rate-limiting step for lymph node metastasis.     

Dopamine-beta-hydroxylase in human lymph

The dopamine-β-hydroxylase (DBH) activity in human lymph was examined. In 14 of 16 persons DBH activity could be detected in the lymph taken from the right lymphatic duct or the thoracic duct. There was a significant (0.05 > p > 0.01) correlation (r = 0.61) between the DBH activity in lymph and serum. The ratio between the levels of DBH in lymph and serum was less than that of the albumin levels (0.36 ± 0.06 and 0.66 ± 0.05).     

Lymphoscintigraphic monitoring of the lower limb lymphatic system response to bone fracture and healing

Closed bone fractures, and torn muscles and tendons are "internal wounds". What kind of reaction do they evoke in the local and systemic immune system? Cellular debris of damaged tissue and extravasated blood cells are removed by scavenger cells. They are transported via lymphatics to the lymph nodes. There elimination of self antigens takes place. Clinically, no enlargement of lymph nodes is observed after closed fractures and soft tissue damage. The question arises whether there is really no enlargement of regional lymph nodes, in other words, no reaction to damaged cell antigens. This question was studied by using lymphoscintigraphy to visualize lymphatics and lymph nodes draining the site of closed bone fracture. The lymphoscintigraphic pictures of two groups of patients, those with a rapid noncomplicated healing of leg fractures, and those with protracted healing and undergoing surgical reconstructions, were evaluated. The surface area of lymphatic pathways and inguinal lymph nodes on the injured and contralateral normal limb were measured. Enlarged superficial lymphatics and inguinal lymph nodes were found in limbs with healed bone fractures, and decreased inguinal lymph nodes and visualization of deep lymphatics and popliteal nodes in the majority of patients with nonhealing fractures. There was a lack of correlation between age of patients, duration of healing, and surgical interventions and the lymphoscintigraphic changes. These findings suggest that the fracture gap tissue is a dominant source of signals to the lymph nodes, releasing cellular and humoral regulatory factors. Taken together, there is a strong immune reaction of lymph node to the fracture, although it cannot be recognized clinically.     

Contractile activity of pectoral channel and lymph nodes in rats under antiorthostatic influences

It has been established that during short term body antiorthostatic positions lymphatic system carries out compensatory function, unloading the cardiovascular system from redundant exercise. The contractile activity of pectoral channel and lymphatic nodes in weightlessness (antiorthostatic influence) was studied. The functional role of neck lymph nodes is expressed by depositing of liquid during redistribution of the blood. The atony of neck lymph nodes as well as loss of pectoral channel and lymph nodes decrease the receptor sensitivity of vessels and nodes to the action of vasoactive substances.     

Intestinal ischemia-induced changes in the activity of NADPH-d and energy metabolism enzymes in rat myenteric and splanchnic plexus

The changes in the activity of NADPH-d and energy metabolism enzymes, lactate dehydrogenase (LDG) and succinate dehydrogenase (SDG), in the neurons of splanchnic and myenteric plexus (SP and MP, respectively), induced by 1-h-long ischemization of a part of the small intestine, were studied using cytophotometric technique; the measurements were performed under conditions when synthesis of nitric oxide (NO) was either blocked or activated. The activity of NADPH-d, SDG, and LDG in MP neurons was shown to be enhanced by ischemia. In SP neurons, the LDG activity increased, while the NADPH-d and SDG activities did not change. The blockade of NO synthesis with nitro-L-arginine methyl ester was followed by a decrease in the NADPH-d level in SP and MP neurons, but was maintained at a level lower than the control one during ischemia. Administration of L-arginine, the NO precursor, increased NADPH-d activity in MP and SP neurons, while at ischemization of the intestine this activity remained at a level higher than in the control. It is concluded that NO-ergic mechanisms (mostly at the MP level) are significantly involved in regulation of the functions of the small intestine in ischemia.