Activity of several carbohydrate exchange enzymes in the lymph during fever reaction

This work includes a comparative study of aldolase enzyme activity, LDG and its isoenzymes in thoracic duct lymph and blood serum of rabbits in fever reaction dynamics of various duration. Irrespective of its duration the fever was followed by a significant activation of investigated enzymes in the body liquids. However, in many day fever reaction a rise of LDG level in the lymph was more prolonged then 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.     

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)     

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.     

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.     

Amino acid composition of the lymph and blood in fever reactions of different durations

The amino acid composition of the thoracic duct lymph and blood following fever reactions (FR) of various duration has been studied in the experiments on rabbits. The experiments have shown that some changes in protein metabolism in FR are primarily manifested in amino acid lymph composition. Irrespective of FR duration the amino acid content in the lymph almost always increases, which confirms the opinion concerning catabolic process predominance in protein metabolism during FR. The lack of "cumulative" conditions in the blood, amino acid dilution in the large volume of the circulating blood, as well as the increase of vascular permeability during FR cause less marked increase in amino acid content in the blood than in the lymph.     

Role of the lymphatic system in the transport of biologically active substances in shock of various etiologies

The content of histamine, serotonin, adrenaline and noradrenaline in the thoracic and lymphatic duct lymph and blood as well as absolute quantity of lymph transported biogenic amines and mediators into the general circulation were studied on 68 dogs during anaphylactic and endotoxin shock (ASh and ESh, respectively). Both ASh and ESh were accompanied by considerable changes in the hemodynamics, lymph circulation, content of biologically active substances in lymph and blood and in their lymph transport to the blood stream. The most profound and early changes in the content of biologically active substances during ASh and ESh were found in the lymph, which shows an important role of the lymphatic system in their resorption and transport from organs and tissues into the general circulation.     

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.     

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.     

Anatomy and topography of the common iliac lymph nodes in human beings in the 1st period of maturity

The investigation of common iliac lymph nodes has been performed in 20 corpses of the first mature age of both sex (5 male and 5 female corpses) of persons died from causes not connected with the lymphatic system diseases, the lower extremities and the pelvic organs. The common iliac lymph nodes with their afferent and efferent lymphatic vessels are revealed by means of interstitial injection into the lower extremities and the pelvic organs and with direct injection into the lymphatic vessels. The form, amount, size and topography of the common iliac lymphatic vessels have been studied. The lymphatic vessels, that go from certain body parts and organs to various subgroups of the common iliac lymph nodes, as well as the lymphatic vessels that connect the nodes both within the subgroup and also between the subgroups. The amount and size of the lymphatic nodes of the lateral subgroup predominate over the nodes of other subgroups of the common iliac lymph nodes; the amount of the common iliac lymph nodes predominates in men, and their size--in women. Amount of these nodes in the right and their size in the left predominate in both sex. Among the common iliac lymph nodes there are no teniform nodes, and efferent lymphatic vessels of the lateral and medial subgroup of the common iliac lymph nodes in 15% of cases run towards the lumbar nodes in the opposite side.     

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.     

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.     

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.     

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.     

Local lymphogenic migration pathway in normal mouse spleen

Although the immunological and hemodynamical significance of the spleen is of great importance, few reports detail the lymphatic vessels in this organ. We have used an immunohistochemical three-dimensional imaging technique to characterize lymphatic vessels in the normal mouse spleen and have successfully demonstrated their spatial relationship to the blood vascular system for the first time. Lymphatic markers, such as LYVE-1, VEGFR-3, and podoplanin, show different staining patterns depending on their location in the spleen. LYVE-1-positive lymphatic vessels run reverse to the arterial blood flow along the central arteries in the white pulp and trabecular arteries and exit the spleen from the hilum. These lymphatic vessels are surrounded by type IV collagen, indicating that they are collecting lymphatic vessels rather than lymphatic capillaries. Podoplanin is expressed not only in lymphatic vessels, but also in stromal cells in the white pulp. These podoplanin-positive cells form fine meshworks surrounding the lymphatic vessels and central arteries. Following intravenous transplantation of lymphocytes positive for green fluorescent protein (GFP⁺) into normal recipient mice, donor cells appear in the meshworks within 1 h and accumulate in the lymphatic vessels within 6 h after injection. The GFP⁺ cells further accumulate in a draining celiac lymph node through the efferent lymphatic vessels from the hilum. These meshworks might therefore act as an extravascular lymphatic pathway and, together with ordinary lymphatic vessels, play a primary role in the cell traffic of the spleen, additional to the blood circulatory system.     

Individual and sexual characteristics of the common iliac lymph nodes in elderly persons

The common iliac lymph nodes (CILN) have been investigated on 24 preparations from corpses of elderly persons (5 male and 7 female corpses), died from the causes not connected with the lymphatic system diseases, lower extremities and pelvic organs. The CILN with their afferent and deferent lymphatic vessels are revealed by means of interstitial injection into the lower extremities and pelvic organs, as well as by means of direct injection into lymphatic vessels. The form, amount, size and topography of CILN are studied. Lymphatic vessels, running from certain parts of the body and organs to various subgroups of CILN are described, as well as lymphatic vessels, connecting the nodes both within each subgroup and between the subgroups. There is a tendency in prevalence of amount and size of the lateral subgroup of the lymph nodes over the nodes of other subgroups of CILN; tendency in prevalence of amount of the lymph nodes in men, and their size--in women; prevalence of amount of right CILN and their size in the left--in persons of both sex; in 70% of the cases the amount of afferent lymphatic vessels to CILN prevails over that of the deferent lymph nodes.     

Coordinated regulation of lymph node vascular-stromal growth first by CD11c+ cells and then by T and B cells

Lymph node blood vessels play important roles in the support and trafficking of immune cells. The blood vasculature is a component of the vascular-stromal compartment that also includes the lymphatic vasculature and fibroblastic reticular cells (FRCs). During immune responses as lymph nodes swell, the blood vasculature undergoes a rapid proliferative growth that is initially dependent on CD11c(+) cells and vascular endothelial growth factor (VEGF) but is independent of lymphocytes. The lymphatic vasculature grows with similar kinetics and VEGF dependence, suggesting coregulation of blood and lymphatic vascular growth, but lymphatic growth has been shown to be B cell dependent. In this article, we show that blood vascular, lymphatic, and FRC growth are coordinately regulated and identify two distinct phases of vascular-stromal growth--an initiation phase, characterized by upregulated vascular-stromal proliferation, and a subsequent expansion phase. The initiation phase is CD11c(+) cell dependent and T/B cell independent, whereas the expansion phase is dependent on B and T cells together. Using CCR7(-/-) mice and selective depletion of migratory skin dendritic cells, we show that endogenous skin-derived dendritic cells are not important during the initiation phase and uncover a modest regulatory role for CCR7. Finally, we show that FRC VEGF expression is upregulated during initiation and that dendritic cells can stimulate increased fibroblastic VEGF, suggesting the scenario that lymph node-resident CD11c(+) cells orchestrate the initiation of blood and lymphatic vascular growth in part by stimulating FRCs to upregulate VEGF. These results illustrate how the lymph node microenvironment is shaped by the cells it supports.