Effect of helium-neon laser rays on the processes of postradiation recovery in the skeletal muscles of old rats

The present experiments were conducted to determine the stimulant effect of helium-neon laser on the postradiation recovery in irradiated uninjured skeletal muscle of rats aged 2-2.5. This was indicated by a restored ability of the muscle for posttraumatic regeneration. The both hind rat legs were exposed to local irradiation of 20 Gy and following laser therapy (8-9 procedures at 3 min each, impulsive or continuous one). Then both musculus gastrocnemius were cut across 30 days after irradiation. It was shown that laser therapy employed before injury of the irradiated muscle accelerated fibrin resorption and improved connective tissue, but slightly stimulated muscular tissue. Impulse laser therapy was more favourable for state of skin and healing of the skin wound after irradiation.     

Immunohistochemical analysis of the distribution of type VI collagen in the lingual mucosa of rats during the morphogenesis of filiform papillae

Iwasaki, S., Yoshizawa, H. and Aoyagi, H. 2012. Immunohistochemical analysis of the distribution of type VI collagen in the lingual mucosa of rats during the morphogenesis of filiform papillae. —Acta Zoologica (Stockholm) 93 : 80–87. We examined the distribution after immunostaining of immunofluorescence of type VI collagen, differential interference contrast (DIC) images, and images obtained using confocal laser‐scanning microscopy in transmission mode, after toluidine blue staining, during morphogenesis of the filiform papillae, keratinization of the lingual epithelium and myogenesis in the rat tongue on semi‐ultrathin sections of epoxy resin‐embedded samples. Immunoreactivity specific for type VI collagen was dispersed over a relatively wide range of connective tissue in the mesenchyme of fetuses on day 15 after conception (E15), at which time the lingual epithelium was composed of one or two layers of cuboidal cells and the lingual muscle was barely recognizable. Slight immunoreactivity specific for type VI collagen was scattered within the lamina propria in fetuses on E17 and on E19, and immunoreactivity was relatively distinct on the connective tissue around the lingual muscle during myogenesis. In fetuses on E19, the epithelium was already stratified squamous. At postnatal stages from P0 to P14, keratinization of the lingual epithelium advanced gradually as morphogenesis of the filiform papillae proceeded during postnatal development. In newborns on P0, myogenesis of the tongue was almost completed. The intensity of immunoreactivity specific for type VI collagen at postnatal stages was mainly restricted on the endomysium and perimysium around the lingual muscle, while scant immunoreactivity was evident in the connective tissue in the lamina propria. Immunoreactivity around the fully mature lingual muscle on P7 and P14 was weaker than that on E19 and P0. Thus, type VI collagen appeared in the connective tissue that surrounded the lingual muscles such as the endomysium and perimysium, in parallel with changes in extracellular components during myogenesis of the tongue.     

Morphofunctional characteristics of the thymus and muscle regenerates under laser irradiation and alloplasty of adult muscle tissue in the area of trauma

The effect of muscle tissue alloplasty and He-Ne laser radiation on the skeletal muscle regeneration and thymus function was studied. Allogenic muscle tissue was implanted from an adult rat. Without laser irradiation (series 1), initial enhancement of thymus recovery observed on day 7 after alloplasty (a characteristic stress response to operation) was followed by gradual destructive changes in the thymus tissue. On day 30 after alloplasty, connective tissue developed in the implantation area in muscle regenerates, and the muscle tissue accounted for 64 ± 2%. Implantation of unirradiated allografts into the muscles of recipient rats preirradiated with a He-Ne laser (series 2) resulted in a nearly complete destruction of the thymus and suppression of its function; the mitotic index of thymocytes was low. These changes were observed throughout the experiment starting immediately after the operation. In this case, the allogenic transplant retained the ability to develop: the 30-day repairing muscles consisted of 71 ± 2% of muscle tissue. When an allograft preirradiated with a He-Ne laser was implanted into unirradiated rats (series 3), thymus destruction at the beginning of the postoperative period was much less significant than in series 2 but more pronounced than in series 1. Then, thymus recovered more rapidly, the allogenic transplant was resorbed, and the muscle tissue in the regenerates accounted for 62 ± 3%.     

Plasticity of allogeneic muscle tissue implanted into a traumatized rat muscle on exposure to He-Ne laser light

The combined effect of muscle tissue alloplasty and He-Ne laser light on the regeneration of a traumatized skeletal muscle was studied. In implantation of untreated allogeneic tissue into the region of transverse cutting of the gastrocnemius muscle exposed to laser light in vivo before surgery, the muscle regeneration in recipient rats was successful. Active regeneration of the allogeneic tissue and the muscle tissue of the recipient rat was noted. In implantation of laser-exposed allogeneic tissue into the region of trauma of unexposed gastrocnemius muscle, the muscle regeneration was less successful. The alloplastic implant was replaced by connective tissue and prevented the growth of the muscle tissue of the recipient rat in the defective zone. Implantation of laser-exposed allogeneic muscle tissue into the region of trauma of the muscle x-irradiated to a dose of 20 Gy enhanced destructive processes in it.     

Reaction of vessels of the microcirculatory bed of subcutaneous tissue to exposure to an argon laser beam

The effect of argon laser irradiation on the microvessels of the subcutaneous fat tissue of rabbit's ear placed into Clark's transparent chamber was studied. Capillaries and venules were found to be sensitive to argon laser irradiation; functional disturbances of microcirculation were most distinct. Following laser irradiation an active reconstruction of the microcirculatory bed with the redistribution of the blood stream took place along with destruction of the vascular wall and perivascular inflammation.     

Reparation of the vascular wall after laser recanalization of the artery (characteristics of changes in the vascular wall at a late period after laser recanalization)]

Chronic experimental studies were carried out on 9 mongrel dogs. To study the reparation processes in the arterial wall after laser intervention, arteries of different anatomic zones were irradiated with laser: descending and abdominal aorta, iliac, femoral and axillary arteries. To make a laser trauma, two types of laser catheters were used: a) with the light guide with diameter of 400 and 600 microns; and b) with a quartz tip at the end. The laser light source was a laser of Cu vapors, and ND; YAG laser. Laser energy was transmitted in the form of individual impulses with the duration of 2-5 sec, and power of 8-10 watt. Histological studies of the preparations of the irradiated arterial wall were performed at periods: 1-3-7-16-28 days, and 2-8 months. Characteristic signs of the vascular wall reparations are given. The analysis of the experimental results showed that by the 28-th day after laser trauma, on the site of laser irradiation a connective tissue scar was formed substituting the defect in the vascular wall and covered from the lumen side with a thin continuous layer of endothelium without any signs of inflammatory infiltration. The development of intimal hyperplasia is not excluded, as well as of inflammatory reaction with the following thrombotic occlusion of the artery lumen. The study results can be used in clinical practice in order to determine the optimum tactics and performance of laser recanalization of the artery, as well as the treatment after the laser intervention.     

日本沼虾消化道形态和组织学特点

应用石蜡切片和扫描电镜技术对日本沼虾消化道进行了研究。结果表明,食道壁向腔内形成四个纵突,食道上皮由单层柱状细胞构成,上皮下的结缔组织中具有放射肌和皮肤腺,环肌层近于连续。食道和胃连结处的管腔背方具食道瓣,胃内具胃磨、滤器和滤沟等结构,胃的组织学结构中除无皮肤腺分布外与食道相似。中肠较长,约占整个消化道的７１７％,具一对中肠前盲囊。中肠上皮细胞大致有两种类型,基膜着色深,环肌层连续,纵肌成束分散排列。后肠为一短管,管腔呈迷路状,其中部为一球形膨大的直肠。后肠的组织学结构与前肠相似。     

Regeneration of the x-ray irradiated gastrocnemius muscle in old rats after stimulation

In the present study the implantation of nonexposed muscle tissue to the site of injury in irradiated musculus gastrocnemius and following laser therapy were applied in order to stimulate this muscle's posttraumatic regeneration in old rats. It was shown that a far larger amount of functionally active muscle tissue was formed at the site of injury compared with rats received laser therapy alone or the ones which were only implanted nonexposed minced muscle tissue. The muscle tissue consisted of muscle fibers which originated from the grafted pieces of nonexposed skeletal muscle and the ones produced by myofibers of muscle stumps recovered after irradiation. The connective tissue developed more evenly. The formation of adipose tissue was not observed at the site of injury. Moreover, the skin wound healing and the hair growth were stimulated as well.     

Smooth muscle actin expression during rat gut development and induction in fetal skin fibroblastic cells associated with intestinal embryonic epithelium

Abstract. Cytodifferentiation of smooth muscle cells has been analyzed immunocytochemically during rat intestinal development and in chimaeric intestines by using monoclonal antibodies reacting specifically with smooth muscle actin species ( CGA7 [10] and anti-α SM-1 [40]). As development proceeds, the various intestinal muscle layers differentiate in the following order: (1) cells expressing smooth muscle actin appear within the mesenchyme of the 15-day fetal rat intestine, in the circular muscle-forming area, the differentiation of cells in the presumptive longitudinal muscle layer starting with a 48-h delay; (2) smooth muscle fibers appear within the connective tissue core of the villi shortly after birth, in parallel with a progressive formation of the muscularis mucosae, which becomes clear-cut only in the course of the 2nd week after birth; (3) a distinct cell layer in the innermost part of the circular muscle layer arises during the perinatal period. Thereafter, the fluorescence pattern remains unchanged until the adult stage. Chimaeric intestines were constructed by the association of 14-day fetal intestinal epithelium and cultured fetal rat or human skin fibroblasts. These fibroblastic cells did not express actin at the time at which they were associated. The immunocytochemical analysis of smooth muscle actin in the hybrid intestines, which had developed as intracoelomic grafts for 12 days, revealed that the skin fibroblastic cells had been induced by the intestinal epithelial cells to differentiate into smooth muscle cells. Such a result was also obtained with allantoic endoderm. It was not obvious in cocultures of intestinal epithelium with skin fibroblastic cells. However, when intestinal epithelial cells were cocultured with intestinal mesenchymal cells, actin expression was stimulated in the latter cell population.     

Expression of T-cadherin in the rat carotid artery wall after balloon injury and in different rat organs

T-cadherin is an unusual glycosilphosphatidylinositol (GPI)-anchored member of the cadherin family of cell adhesion proteins. In contrast to classical cadherins, tissue distribution of T-cadherin so far remained unknown. We examined tissue distribution of T-cadherin in rats using Western blotting and immunohistochemical method. Our results show that T-cadherin is expressed in all types of muscles (cardiac, striated, and smooth muscles), in brain neurons, and spinal cord, in the vessel endothelium, at the apical pole of intestinal villar epithelium, in the basal layer of skin, and eosophagal epithelium. Blood-derived and lymphoid cells as well as connective tissue were T-cadherin-negative. The highest level of T-cadherin expression was revealed in the cardiovascular system. Although T-cadherin was detected in smooth muscle cells, its role in the intimal thickening and restenosis is not known. We examined T-cadherin expression within 1-28 days after balloon injury of rat left carotid arteries. T-cadherin expression was valued immunohistochemically with semiquantitative method. In uninjured arteries, T-cadherin was expressed in endothelial (vWF-positive) cells, and smooth muscle (alpha-actin-positive) cells (SMCs). After denudation of arterial wall, T-cadherin was present both in the media and neointima. We revealed dynamics of T-cadherin expression in the media of injured artery: an essential increase being registered at the stage of cell migration and proliferation in the media and neointima (1-7 days), followed by its decrease to the baseline level (10-28 days). The high upregulation of T-cadherin expression in the media and neointima during migration and proliferation of vascular cells after vessel injury enables us to suggest the involvement of T-cadherin in vessel remodeling after balloon catheter injury.     

Skin and subcutaneous fat morphology alterations under the LED or laser treatment in rats in vivo

The main objective of this work is to quantify the impact of photodynamic/photothermal treatment by using visible LED and NIR laser irradiation through the skin of subcutaneous fat in vivo followed up by tissue sampling and histology. The optical method may provide reduction of regional or site‐specific accumulations of abdominal or subcutaneous adipose tissue precisely and least‐invasively by inducing cell apoptosis and controlled necrosis of fat tissue. As photodynamic/photothermal adipose tissue sensitizers Brilliant Green (BG) or Indocyanine Green (ICG) dyes were injected subcutaneously in rats. The CW LED device (625 nm) or CW diode laser (808 nm) were used as light sources, respectively. Biopsies of skin together with subcutaneous tissues were taken for histology. The combined action BG‐staining and LED‐irradiation (BG + LED) or ICG‐staining and NIR‐laser irradiation (ICG + NIR) causes pronounced signs of damage of adipose tissue characterized by a strong stretching, thinning, folding and undulating of cell membranes and appearance of necrotic areas. As a posttreatment after 14 days only connective tissue was observed at the site of necrotic areas. The data obtained are important for safe light treatment of site‐specific fat accumulations, including cellulite. This work provides a basis for the development of fat lipolysis technologies and to move them to clinical applications. Schematics of animal experiment.      

Conjugated effects of thyroxine and X-rays on the intestinal wall of Alytes obstetricans larvae (Anuran amphibian)

Summary The conjoined effects of thyroxine and X-rays on the intestinal wall were studied using Alytes obstetricans tadpoles in premetamorphosis.Thyroxine alone induces degeneration of the larval epithelium (primary epithelium) and its replacement by a secondary epithelium. The latter is derived from stem cells via the development of islets.In animals submitted to irradiation only, many of these stem cells showed signs of necrosis.In irradiated larvae treated with thyroxine, the secondary epitheliocytes were rare and never formed islets. Radioautographic observations confirmed their very low proliferation rate. Contrary to what was observed in the hormone treated larvae, cell fragments of the primary epithelium were extruded in the connective tissue, and phagocytes appear to infiltrate the epithelium.In animals treated with thyroxine and later submitted to irradiation, islets of secondary epitheliocytes developed while some cells degenerated. There again, the phagocytes were noted in both the connective tissue and the epithelium.     

AUTORADIOGRAPHIC LOCALIZATION OF NEW RNA SYNTHESIS IN HYPERTROPHYING SKELETAL MUSCLE

Work-induced growth of rat soleus muscle is accompanied by an early increase in new RNA synthesis. To determine the cell type(s) responsible for the increased RNA synthesis, we compared light autoradiographs of control and hypertrophying muscles from rats injected with tritiated uridine 12, 24, and 48 h after inducing hypertrophy. There was an increased number of silver grains over autoradiographs of hypertrophied muscle. This increase occurred over connective tissue cells; there was no increase in the number of silver grains over the muscle fibers. Quantitative studies demonstrated that between 70 and 80% of the radioactivity in the muscle that survived fixation and washing was in RNA. Pretreatment of the animals with actinomycin D reduced in parallel both the radioactivity in RNA and the number of silver grains over autoradiographs. Proliferation of the connective tissue in hypertrophying muscle was evident in light micrographs, and electron micrographs identified the proliferating cells as enlarged fibroblasts and macrophages; the connective tissue cells remained after hypertrophy was completed. Thus, proliferating connective tissue cells are the major site of the increase in new RNA synthesis during acute work-induced growth of skeletal muscle. It is suggested that in the analysis of physiological adaptations of muscle, the connective tissue cells deserve consideration as a site of significant molecular activity.     

Shell attachment in the pond snail Lymnaea stagnalis (L.)

The attachment of the body of the snail Lymnaea stagnalis to the shell was studied by histochemistry and light and electron microscopy. Muscles of the body wall insert into the connective tissue by way of long thin projections of sarcolemma. The muscle cells end under the basement membrane of a specialised area of the epidermis, the adhesive epithelium. The cells of this epithelium are filled with microfilaments and possess characteristic knob-like microvilli. The epithelium is attached to the shell by way of an adhesive substance containing proteins and mucopolysaccharides.This research was made possible by a grant from the Netherlands Organization for Pure Research (Z.W.O.)     

Pro-inflammation and pro-fibrosis factors were highly induction in heart tissues of carotid arteries balloon-injured animal model

To investigate the changes of cardiomyocyte inflammation and fibrosis factors in heart of carotid artery balloon injury inflammatory rat model. Using rat carotid artery balloon injury model to detect left ventricular characteristics at 2 h, 2 days and 14 days after surgery using hematoxylin-eosin (H&E) gross stain, Masson's trichome stain and Western blot analysis for inflammatory and fibrosis-induced factors, tumour necrosis factor α (TNFα), JNK1, P38α, connective tissue growth factor (CTGF), SP1 and transforming growth factor β (TGFβ) protein expressions. The rat carotid arteries were injured after 2 h, 2 days and 14 days. Balloon-angioplasty to H&E stain results showed the increasing trend of left ventricular wall at 2 h and 2 days; then, the left ventricular wall became thinner, and the left ventricular chamber became enlarged and dilated after 14 days of carotid artery balloon injury. In addition, the Masson's trichome stain results showed that the left ventricular section has fibrosis-related blue staining (collagen) at 2 and 14 days after rat carotid artery balloon injury, and became even more severe at 14 days. Furthermore, we observed the protein expression level changs, which include TNFα, JNK1, P38α, CTGF, SP1 and TGFβ using Western blotting assay. All proteins were induced at 2 h, 2 days and then reached the maximal level at 14 days. The vessel inflammation was associated with cardiac inflammatory and fibrosis effects during or after carotid artery balloon injury.     

Microsurgical anastomosis of rat carotid arteries with the CO2 laser

In order to further evaluate the role of lasers in microvascular tissue closure, we modified an existing CO2 surgical laser (Xanar XA-20) by adding a partially reflecting mirror to attenuate the beam. This allowed the laser to operate at an output of approximately 100 mW, which was appropriate to achieve microvascular closures. In each of 43 rats, one carotid artery was transected and then anastomosed with standard suture technique with 10 to 12 simple interrupted sutures of size 10-0 Ethilon nylon suture (Ethicon, Inc.). The opposite carotid in each rat was anastomosed by the placement of three stay sutures followed by the application of laser irradiation to the tissue between the stay sutures at 90 to 100 mW, spot size of 0.2 mm, pulse duration 0.2 seconds, approximately 20 to 30 pulses per anastomosis. In vivo test periods were 1 hour, 1 day, 3 days, 7 days, 10 days, 14 days, 28 days, 91 days, and 180 days. All anastomoses were evaluated for patency, and selected samples were utilized for light microscopy, and mechanical testing (intraluminal pressure raised to 300 mmHg). It was determined that similar patency rates and slightly faster time to perform the same procedure could be achieved with the use of the low-powered CO2 laser. However, histologic evidence of significant medial damage raises concern about the long-term risk of a higher aneurysm rate. Vessel damage and the lack of simple intraoperative methods to verify the quality of the laser technique restrict these authors from advocating the clinical introduction of the procedure until further advances are made.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effect of low-intensity He-Ne laser irradiation on rat mesenteric microcirculation

Changes in microcirculation during low-intensity He-Ne laser irradiation were studied by measurement of blood vessels and lymphangions' diameter in anesthetized rat mesentery using method of videomicroscopy in vivo. We demonstrated that the red coherent light induced a significant increase in contractility of vessels' smooth muscle cells. The flow decreased in the mesenteric microvessels during H2O2-mediated oxidant stress was restored after He-Ne laser irradiation.     

Origin and distribution of VIP (Vasoactive Intestinal Polypeptide)-nerves in the genito-urinary tract

Summary VIP (Vasoactive Intestinal Polypeptide)-immunoreactive nerves were found throughout the genito-urinary tract of the cat; they were less numerous in the guinea pig and in the rat. In the cat, VIP nerves were particularly numerous in the neck of the urinary bladder and proximal urethra, in the uterine cervix and in the prostate gland. The nerves were found in smooth muscle, around blood vessels and in the connective tissue immediately beneath the epithelium. Ganglia were found below the trigonum area of the bladder, in the wall of the proximal urethra, and in paracervical tissue. VIP-immunoreactive nerve cell bodies occurred in all these ganglionic formations. These ganglia probably represent the origin of the VIP nerves of the genital tract since their removal in the female cat greatly reduced the VIP nerve supply. Transection of the hypogastric nerves had no overt effect. Transection of the cervix eliminated the VIP nerves above the level of the lesion, except those in the ovaries, supporting the view that the VIP nerves of the uterus and the oviduct are derived from a paracervical source.     

Early development and pathogenesis of Lobatostoma manteri Rohde (Trematoda: Aspidogastrea)

The prosobranch Planaxis sulcatus is reported as a new natural host of Lobatostoma manteri at Heron Island, Great Barrier Reef. Planaxis sulcatus and Cerithium moniliferum were experimentally infected with large numbers of eggs. The larvae hatch in the stomach and migrate immediately along the ducts of the digestive gland into the digestive follicles. The larvae feed on the secretion and probably epithelial cells of the follicles. The acetabulum is used for adhesion to the epithelium and contributes to its erosion. In heavily infected snails, the digestive follicles disappear gradually and the larvae live in cavities lined by a flattened epithelium, parts of which show secretory activity. In snails dissected 47–49 and 65–66 days after infection, the cavities are fused, forming several large spaces which communicate with each other; only small parts of the epithelium are still secretory. Concentrations of amoebocytes occur in the walls of the digestive gland and in the wall between digestive gland and stomach of infected Planaxis. Some young worms were found in the stomach of Planaxis. No tissue reactions were seen around the stomach except in the wall between digestive gland and stomach. In Cerithium with 65–67 days old infection, the cavities contain much detritus and disintegrating cells, the epithelium is practically non-secretory and surrounded by loose connective tissue. In larvae with a body length of approximately 0·5–0·6 mm, the acetabulum begins to divide into alveoli and its anterior end grows forward; the anterior alveoli gradually increase in size and new alveoli are formed in the posterior undivided zone. In two specimens of approximately 1·3 mm body length, the whole adhesive disk was divided into half the number of alveoli usually found in adults. Allometric shifts during growth of the worms are analysed.