Tunica Layer Research Articles

Could Testicular Tissue Be a New Arena for the Holmium Laser?

Introduction We aimed to observe the effects of holmium:yttrium-aluminum-garnet (Ho:YAG) laser on testicular tissue. Methods An ex vivo experiment was conducted using calf testicles. A 100 W laser generator with broad-spectrum settings of 10-80 W, 20-40 Hz, and 0.5-2 J, with a medium pulse duration, was tested. The laser effects on testicular tissues with and without the tunica layer were evaluated histopathologically by calculating the incision depth (ID), vaporization area (VA), coagulation area (CA), and total laser area (TLA=VA+CA) of the specimens. Results A total of 48 experiments were conducted.In testicular tissue without a tunica layer, the highest mean ID was determined at 1 J-20 Hz (0.247±0.0208 mm) and with a tunica layer at 2 J-40 Hz (2.673±0.032 mm). In the testicular tissue without a tunica layer, the highest mean VA was determined at 1.5 J-40 Hz (0.029±0.0016 mm2)and in tissue with a tunica at 2 J-40 Hz (6.173±0.114 mm2). The highest mean TLA in tissue without a tunica was detected at 2 J-20 Hz (0.038±0.0008 mm2)and in tissue with a tunica at 2 J-40 Hz (7.292±0.07 mm2). The mean ID, VA, CA, and TLA values of all the power outputs used were found to be statistically significantly higher in the testicular tissue with the tunica layer than in that without it (p<0.001). Conclusion The Ho:YAG laser has different effects on testicular tissue with and without a tunica layer. In testicular tissue without a tunica, the laser's effect was minimal on the surrounding tissue, especially in terms of the ID, VA, and TLA.This minimal effect of the laser can be an advantage in testicular surgery procedures such as testis-sparing surgery (TSS) or testicular sperm extraction (TESE).

Meta-Topolin enhanced direct shoot organogenesis and regeneration from leaf explants of Coleus forskohlii (Willd.) Briq

A novel and effective direct shoot regeneration technique from stomatal complexes was developed for a highly sought after medicinal plant Coleus forskohlii. Meta-Topolin was more efficient than 6-benzylaminopurine (BAP) in promoting direct organogenesis from leaf surface. Highest direct organogenesis was achieved from in vitro leaf explants on Murashige and Skoog (MS) medium augmented with meta-Topolin at the concentration of 2.0 mg/L. The regenerated leaves were fixed at weekly intervals and sectioned longitudinally to evaluate the anatomical developments. The results of light microscopic analysis suggested that the in vitro developed stomatal complexes showed new altered cell divisions, after a week of incubation, and the rapid periclinal divisions resulted in the formation of a meristematic dome. Distinct regions of meristematic activity were visible in the second week characterized by promeristems with small and densely stained parenchymatous cells. Promeristems lead to the development of a distinct tunica layer, which grew into shoot-like protuberances converting into adventitious shoots after 4 weeks of incubation. A clear three phases of shoot organogenesis viz. induction (7 days), initiation and organization (14 days), and shoot formation (28 days onwards) were identified. The shoots were proliferated (21.8 shoots/ single leaf explant) and elongated (5.0 cm length) using half-MS medium fortified with 0.5 mg/L mT and indole-3 acetic acid (IAA) at 0.1 mg/L. Notable rhizogenesis of the elongated shoots was achieved when cultured on half-MS medium containing indole-3 butyric acid (IBA) at the concentration of 1.5 mg/L. About 92.0% of the plantlets survived upon field transfer. Start Codon Target (SCoT) markers were employed to examine the regenerated plants’ genetic uniformity with the mother plant, similar banding patterns confirming the genetic uniformity. Hence, the developed protocol of direct in vitro regeneration of C. forskohlii using mT can be used for the commercial production.

Fiber-optic laser energy for pulmonary artery denervation: results of an experimental study

Abstract Background/Introduction Pulmonary artery denervation (PADN) is an interventional procedure aiming to modulate and correct pulmonary artery (PA) pressure. New technologies and energy sources are being tested currently. However, the effects of laser energy on the PA wall are unknown. Purpose To assess the acute effects and impact of laser energy for PADN procedure in normotensive sheep by applying different power settings and session times. Methods A total of 10 normotensive Katumsky sheep were included in the experiment. Percutaneous vascular access was performed under general anesthesia. A fiberoptic open-irrigated non-steerable catheter was introduced via a femoral vein through a steerable sheath. Laser applications were applied in the PA trunk and the proximal areas of the right and left PA under fluoroscopic guidance with a 5mm distance between points in the anterior, posterior and lateral walls. Applications were delivered with power 10–30W, 10–35s in duration; irrigation flow 40ml/min. After the procedure, experimental animals were euthanized and underwent an autopsy. PA samples were obtained regardless of the absence of visible laser-related lesions for histological analysis (hematoxylin staining) and immunohistochemical labelling (S100). Results A total of 108 ablation sessions were performed, 33 in the right PA, 30 in the left PA and 42 in the PA trunk. During macroscopic examination, laser-related lesions described as irregular brown hemorrhage spots and rough defects observed in the PA endothelium were not homogenous in all experimental animals. Thermal injuries either in the left or right lung lobes were identified in 5 (50%) experimental animals when using 30W during 10–20sec. In 5 (50%) sheep no collateral lung injuries were identified when using 10–20W from 20–35s. A total of 64 PA fragments underwent microscopic examination, acute thermal tissue lesions were observed in all experimental animals despite the absence of laser-related lesions in the PA endothelium; dissection, edema, disruption trough tunica layers, hemorrhage and necrosis at different depth walls. The most frequent nerve damage was obtained with 20W ablation: 5/8 PAs vs, 1/6 with 10–15W and 1/14 with 25–30W (P=0.01). At the same time, there was no difference in intima necrosis between the groups (1/6, 2/8 and 4/14 for 10–15, 20 and 25–30W groups, respectively). Conclusion(s) Percutaneous PA laser ablation is feasible, reduction of perivascular nerve expression is seen most frequently, when 20W/20–35 s ablation is performed. PA lesions may differ in depth and characteristics, and perivascular nerve damage might be seen in cases with preserved intima. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): Grant of the Ministry of Science and Higher Education of the Russian Federation

Septicemic Infection Suspect In Jenkins' Whipray Pateobatis jenkinsii (Annandale, 1909): A Case Report

A wild-captive male Jenkins' whipray was found dead in a quarantine tank with a clinical sign before death in the form of decreased appetite for a week. The treatment history was oral administration of enrofloxacin antibiotic tablets. The therapy period lasts for ten days. The last treatment was the administration of Hepavit® (liver extract) and intramuscular injection of enrofloxacin antibiotic. One day before the death, blood was collected and then examined for the hematocrit and some parameters of chemical blood. The results of blood examination were found a decrease in blood urea nitrogen (BUN), alkaline phosphatase (ALP), and alanine aminotransferase (ALT) levels, increased glucose level, decreased total protein and albumin levels, and increased globulin level. Anatomical pathology examination was found lesions on the tail, around the eyes, and claspers. Hemorrhagic lesions were found in the mucous layer of the esophagus, stomach, and spiral colon. The blood clot was found under the tunica layer of testicular organs. The liver is damaged by showing a non-homogeneous coloration, organ thickening, congestion, and fragile consistency. Based on the results of the blood examination and was supported by the results of the anatomical pathology examination after death, it is suspected that the fish died due to the condition of septicemia infection during the previous few weeks.

Morphological description of smell glands in Cyclopes didactylus.

A macroscopic and microscopic study of the mandibular organ of the silky anteater (Cyclopes didactylus) was carried out. The organ extends from below the zygomatic bone line to the middle of the mandible body, between the skin and the masseter muscle, on both sides of the animal. It has an average length of 11.7 mm and a width of 6.3 mm. In the mesoscopic analysis, it was observed that the organ presents in yellowish color due to the high amount of sebaceous content. In the histological analysis, the mandibular organ was observed to be composed of innumerable alveoli of the specialized sebaceous gland, surrounded by a layer of adventitia tunica. Scanning electron microscopy (SEM), revealed an apparent alveolar division with what appeared to be a sulcus at its center. The information here presented regarding the constitution and location of this structure has not been previously explored for other species and differs with respect to other descriptions for anteaters. Based on the present study, it is suggested that the mandibular organ is involved in social interaction in this species.

Increase and Redistribution of Sex Hormone Receptors in Premenopausal Women Are Associated with Varicose Vein Remodelling.

In chronic venous insufficiency of the lower limbs, data show that the clinical manifestation is varicose veins (VVs), and VV epidemiology suggests that sex hormones directly influence disease development through intracellular receptors. This study aimed to determine the presence and localization of oestrogen receptors (ERs), progesterone receptors (PRs), and androgen receptors (ARs) in both healthy and VV wall cells and their relationship with gender. In this study, samples from patients without a history of venous disease (CV) (n = 18) and with VV (n = 40) were used. The samples were divided by gender: CV women (CVw) = 6, CV men (CVm) = 12, VV women (VVw) = 25, and VV men (VVm) = 15. RT-qPCR and immunohistochemical techniques were performed, and increased ER and PR protein expression was found in VVw in all tunica layers. ARs were localized to the adventitial layer in the CV and were found in the neointima in VVs. mRNA expression was increased for ER and PR in VVw. AR gene expression was significantly decreased in VVm. The increase in the number of these receptors and their redistribution through the wall reinforces the role of sex hormones in varicose vein development.

Desarrollo Anatómico de los Meristemas y tallos de Alnus jorullensis H. B. K.

The shoot apex of Alnus jorullensis comprises two well-defined tunica layers and a corpus, which gives rise to foliar primordial and to procambial strands maturing acropetally. The primary shoot shows a eustek within abundant perenchyma consisting of isodiametric cells. Three leaf traces and three leaf gaps are seen at each node. Secondary xylem is made up of abundant, poorly lignified fiber tracheids, with apotracheal wood parenchyma, uniseriate and scantily biseriate xylem rays and apparently mucilaginous secretory cells. Vessel clements range about a mean length of 710 µm and a mean width of 70 µm with end-wall angles averaging 24° 56'. Perforation plates are scalariform, ranging fron 9 to 26 bars (Av. 16.48). In the course of the first year's growth, side-wall pitting changes from the metaxylemic reticulate-scalariform to the secondary opposite, bordered pattern.

A new corporoplasty based on stratified structure of tunica albuginea for the treatment of congenital penile curvature - long-term results.

IntroductionThe aim of the study was to report long–term results of treatment of patients with congenital penile curvature (CPC) with a new corporoplasty based on stratified structure of tunica albuginea, in which corporal bodies are not opened.Material and methodsFrom October 2006 to September 2013, the authors operated on 111 adult men with CPC. Ventral curvature was detected in 65 patients, lateral in 34, and dorsal in 12. Skin was incised longitudinally on convex surface of curvature. In ventral curvature, dorsal neuro–vascular bundles (NVBs) were separated from tunica albuginea and elliptical fragments of external (longitudinal) layer of tunica were excised. The tunica was sutured with absorbable sutures, which invaginated the internal (transversal) layer of tunica. In dorsal curvature, excisions were performed on both sides of the urethra, in lateral curvature – on the convex penile surface.ResultsFollow–up period was from 12 to 84 months. The penis was completely straight in 109 out of 111 patients. In 2 patients (1.8%) recurrent curvature of up to 20 degrees was detected. Redo surgery was done in one individual (0.9%) at patient‘s request. Glandular sensation loss or erectile dysfunction was not detected in any patient during the period of observation.ConclusionsA new operation for correction of CPC, which consists of excision of an elliptical fragment of the external layer of the tunica albuginea and plication of the internal layer gives good short and long–term results. Surgery done without penetrating the corpora cavernosa is minimally invasive, which diminishes the potential risk of complications, especially intra– and postoperative bleeding.

Growth and development of shoot apex in barley I. Morphology and histology of shoot apex in vegetative phase

The vegetative phase of development of the main shoot apex lasts over 5 plastochrons after germination. The endosperm has a sufficient store of nutrition for this period. At the beginning of this phase the apex has a one-layer tunica. The cells of the latter divide above the level of bulge formation for leaf primordia, exclusively anticlinally, although somewhat lower within the leaf bulge periclinal divisions may occur. The cells immediately under the first tunica layer in the apical part grow tangentially to the surface. These cells divide only anticlinally forming gradually the second tunica layer. In the course of the entire phase the shape of the meristeanatic caulis from the tip to the 4th frustum remains unchanged.

Histological study of cartilaginous cells in the body of penis in one-humped camel (camelus dromedarius)

BACKGROUND:Several investigations showed cartilaginouscells in fibrous tissue of the free part of the penis in one humpedcamel. OBJECTIVES: The aim of this study was accurate assessmentof existence of cartilaginous cells in penis shaft of onehumpedcamel. METHODS: Six camel penises from maturedcamels more than 3 years-old were collected from an abattoir.Different specimens were prepared from each penis and kept in10% formalin container for fixation. After passing differentstages of histotechnique methods, several slides were preparedfrom each specimen, stained with Haematoxylin Eosin andstudied. RESULTS: Results showed that the majority of cartilaginouscells were inside the collagen fibers of tunica albuginea andaround corpus cavernosum and corpus spongiosum of penis andtheir distributions were dissimilar in different parts of the penisshaft. This survey further showed that in penis shaft length, themajority of cartilaginous cells were inside tunica albuginea,which is surrounded by corpus spongiosum and particularly, theventral surface of urethra. CONCLUSIONS: The number ofcartilaginous cells decreased gradually from distal extremitytowards the proximal extremity of the body of the penis andincreased gradually from external layer of tunica albugineatowards the internal layer of tunica albuginea and centre ofcorpus cavernosum penis. Existence of cartilaginous cells insidethe leaf tissue of the penis was seen with aging and puberty.

Spatio-temporal expression of PsAPY1 during germination, differentiation, and organogenesis in pea (Pisum sativum L.Var. Alaska)

Apyrase (ATP-diphosphohydrolase, EC 3.6.1.5) catalyses the hydrolysis of nucleoside tri- and di-phosphates to nucleoside monophosphates and orthophosphates. In the present study, the spatio-temporal expression of an apyrase gene (PsAPY1) from pea (Pisum sativum L. Var. Alaska) was investigated during the early stages of germination [10 h after imbibition and 16, 35, 62, and 84 h after sowing (HAS)], using non-radioactive mRNA in situ hybridization. At 10 h after imbibition, the PsAPY1 transcript was visible in traces throughout the shoot and root tissues of the embryo. The transcript accumulation became higher by 16 HAS in the tunica layer of the shoot apical meristem, epidermis, vascular bundle, and root tip. At 35 HAS, expression signals became stronger and more widely distributed in the corpus tissues of the growing shoot apical region and in the root; the highest expression was detected in the epidermis, differentiating vascular bundle, and somewhat less in cortex tissues. By 62 HAS, the expression of PsAPY1 was strongest and extensively distributed in the above mentioned regions. At this time, higher expression was also detected in the zones of epicotyl differentiating into plumule and hook. However, at 84 HAS, the expression of PsAPY1 declined in the shoot and root, but was highly expressed in growing lateral root primordia. Germination and growth of seedlings are the physiologically most important stages in the life cycle of plants. The presence of PsAPY1 during these important stages i.e., germination, differentiation, and organogenesis indicates its essential role in growth and development of plants.

A Comprehensive Protein Expression Profile of Extracellular Matrix Biomaterial Derived from Porcine Urinary Bladder

To generate a comprehensive profile of the protein composition of xenogeneic biomaterial, derived from porcine urinary bladder matrix (UBM). Tunica layers and muscularis mucosa were removed from bladders using mechanical delamination. UBM was prepared using a solution of peracetic acid in ethanol, lyophilized then milled into powder. UBM biomaterial was subjected to tryptic digests and components separated using high-performance liquid chromatography with an ion trap mass spectrometer and identified through databases. A repertoire of 129 proteins with neurotrophic, antiangiogenic and tumor-suppressive activities and those associated with tissue remodeling and wound repair were identified. This study provides the first insight into the complex nature of the UBM and how its application may be tailored for specific applications in regenerative medicine. We propose that the UBM be further investigated for reconstructive and regenerative remodeling of cardiac and dermal tissues, as well as peripheral nerves.

Epithelioid malignant mesothelioma of tunica vaginalis with deciduoid features: An unusual malignancy clinically masquerading an inguinal hernia

Paratesticular/scrotal and inguinal canal mass lesions in elderly patients may pose a diagnostic challenge to both the surgeon as well as the pathologist. In most cases, these represent hernial sacs with their contents, and true neoplasms like lipomas, rhabdomyosarcomas, and fibrous pseudotumors are infrequent. Malignant mesotheliomas arising from the tunica layers are rare cause of inguinal and paratesticular tumors. Herein, we report a case of an elderly patient who presented with an inguinal hernia which pathologically had features of deciduoid malignant mesothelioma.

Histocytological characteristics of Eucalyptus urophylla × Eucalyptus grandis shoot apical meristems of different physiological ages

Histocytological characteristics of Eucalyptus urophylla × Eucalyptus grandis shoot apical meristems (SAMs) were described, comparing five outdoor and in vitro sources of akin genotypes differing in their physiological age. The size and the number of cells of the five zones identified within each SAM, i.e. the two tunica layers (L1 and L2), the central mother cells (CMC), the peripheral zone (PZ) and the combination of these four zones (4CZ) varied according to physiological age and plastochron phase. These five zones were significantly larger with higher numbers of cells for SAMs from mature and juvenile trees than for those from physiologically rejuvenated, in vitro mature and in vitro juvenile plants. However, these origin-related differences were not significant for SAMs in their early plastochron phase, to become obvious in a more advanced plastochron stage. Individual cell and nuclear measurements confirmed the rationale of distinguishing within SAM zones, characterized by specific cell and nuclear sizes liable to vary according to physiological age. The various histocytological investigations carried out established that SAM cell characteristics appeared to be the more reliable indicators of phase change. This was particularly true for the nucleoplasmic ratio and for more qualitative differences observed also at the nuclear level. SAM nuclei of the two in vitro origins were more evenly stained by naphtol blue-black, uniformly light for the juvenile source, whereas the mature source showed also darker nuclei. In contrast, SAM nuclei from outdoor origins had more chromocenters, darker and diffusely spread for the mature source than for the rejuvenated and the juvenile origins, where they were more peripherally distributed and where the nucleoli appeared more clearly. These results were discussed with respect to physiological ageing and in vitro culture influence, and suggest a determining influence of SAM cell nuclei on phase change phenomenon of arborescent species.

The role of tunica vaginalis graft as the dorsal component of urethra in 1-stage and 2-stage hypospadias repair: experimental study in rabbits

Purpose The paucity of tissue for the urethroplasty in complex Hypospadias has opened a discussion and the search for alternative materials. The buccal mucosa is the most popular tissue used to date. We were interested to evaluate the use of tunica vaginalis graft as dorsal component of urethra anastomosed either to normal urethral mucosa, to a penile preputial flap and also as a graft in 2-stage Bracka strategy. Material and Methods We used a rabbit model and created a defect in the penile urethra and developed 3 experiments to study clinical outcome and histological healing of the neourethra with hematoxilin-eosine and Masson's trichrome staining techniques. For each experiment (n=16), 4 subgroups of sacrifice with 4 animals each were defined according to sacrifice date of 2, 4, 8 and 16 weeks. The difference of experiment 1 to 2 was that dorsal tunica vaginalis was anastomosed to normal urethra (E1) and to a transverse preputial flap (E2) whereas in experiment 3 (E3) the tunica vaginalis graft was left exposed to external environment. Results The macroscopical aspect of the surgery was uneventful in E1 and E2, whereas E3 showed an edematous and inflammatory aspect with progressive and spontaneous resolution after 4 weeks. The histological analysis of the graft was very similar in both clinical situations. The aspect of a mesotelial one cellular layer of tunica was progressive replaced by a 4-5 layer cuboid-cells epithelium at 16 weeks, suggesting metaplasia, since no follicles and keratin were seen. Interestingly, the “Bracka” tunica vaginalis surgery had the same histological appearance of dorsal TV urethroplasty when anastomosed either to normal urethra or to a foreskin flap. Conclusions The experimental findings suggest us that tunica vaginalis may have a role in urethral reconstruction when used as a dorsal graft attached to corpora and could be used as an alternative to other recognized options such as buccal mucosa and foreskin.

Influence of clavata3-2 mutation on early flower development in Arabidopsis thaliana: quantitative analysis of changing geometry.

Early development of the flower primordium has been studied in Arabidopsis thaliana clavata3-2 (clv3-2) plants with the aid of sequential in vivo replicas and longitudinal microtome sections. Sequential replicas show that, although there is no regular phyllotaxis in the clv3-2 inflorescence shoot apex, the sites of new primordium formation are, to a large extent, predictable. The primordium always appears in a wedge-like region of the meristem periphery flanked by two older primordia. In general, stages of primordium development in clv3-2 are similar to the wild type, but quantitative geometry analysis shows that the clv3-2 primordium shape is affected even before the CLAVATA/WUSCHEL regulatory network would start to operate in the wild-type primordium. The shape of the youngest primordium in the mutant is more variable than in the wild type. In particular, the shape of the adaxial primordium boundary varies and seems to be related to the shape of the space available for the given primordium formation, suggesting that physical constraints play a significant role in primordium shape determination. The role of physical constraints is also manifested in that the shape of the primordium in the later stages, as well as the number and position of sepals, are adjusted to the available space. Longitudinal sections of clv3-2 apices show that the shape of surface cells of the meristem and young primordium is different from the wild type. Moreover, there is only one tunica layer in both the meristem and in the primordium until it becomes a bulge that is distinctly separated from the meristem. Starting from this stage, the anticlinal divisions predominate in subprotodermal cells, suggesting that the distribution of periclinal and anticlinal cell divisions in the early development of the flower primordium is not directly affected by the clv3-2 mutation.

Quantitative morphological analysis of the myenteric neurons of the ileum in rats under experimental desnutrition

This paper deals with the effects of proteic desnutrition on the morphology, morphometry and density of the neurons of the myenteric plexus. The ileum of 10 rats was used. For this study five rats aging 90 days were fed during 120 days with hypoproteic ration (experimental group) and five rats (control group) received ration with normal proteic level. Segments of the ileum were obtained for the method of whole-mount preparation, stained with Giemsa (Barbosa, 1978) and for histological routine treatment stained with HE. The neurons were clustered in ganglia located between the circular and longitudinal layers of the muscular tunica. Based on the lengths of the major longitudinal and transverse axes, the neurons were selected in three groups: small, medium and large. The neuronal density on the ileum of rats corresponding to an area of 7.08mm 2 was 1,482 and 2,515 neurons on the control and experimental groups respectively. The data suggested that proteic desnutrition did not cause alteration to the neuronal density.

Cambios morfoanatómicos en los ápices del vastago y de la raíz del banano Williams (AAA, Musa sp.) bajo distintas concentraciones de N6-benciladenina

Hoy en día existen escasas evidencias estructurales acerca del crecimiento in vitro de los ápices del vastago (AV) y de la raíz (AR) del banano. Este trabajo tiene como objetivo analizar los cambios morfoanatómicos que ocurren durante el crecimiento del AV, AR y explante del banano Williams bajo distintas concentraciones de N6-benciladenina (BA). Se estudiaron AV de explantes (8 mm X 1.5 mm, abarcando punta del vastago con porción del rizoma) cultivados en 0, 2.5 y 5 mg/L de BA durante cero, tres, seis, nueve y 12 días y, asimismo, la primera raíz de dichos explantes cuando alcanzó 1 a 1.5 cm. Las muestras se seccionaron (10 a 12 µm) y se tiñeron con safraninafast green. El AV presentó forma de domo con organización túnica-corpus (una capa de túnica). Su diámetro fue mayor al cultivarlo con BA (93.75 a 142.05 µm) que sin esta citoquinina (73.87 a 85.23 µm), exceptuando los valores del día 6 (127.84 µm, sin BA). En el explante inicial sin cultivo el diámetro fue de 164.78 µm. En 2.5 mg/ L de BA al día nueve, el AV mostró una zona transicional parecida al cambium. Esta concentración indujo mayor número de brotes por explante (2.19) a los 35 días. El AR en medio sin BA presentó protodermis, meristemo fundamental, procambium, células iniciales y caliptra mientras que con BA se identificaron procambium, meristemo fundamental y caliptra (comprimida). La benciladenina disminuyó la cantidad y la longitud de las raíces, inhibió la producción de primordios de raíces laterales, atrasó el período de emergencia radical y produjo distorsión en su morfoanatomía.

Stem cell renewal and contraction of the tunica media caused by a damaged blood vessel following a thick needle stab

A marked difference in the healing process of the inferior vena cava in rats following a stab with a 17-G (1.48 mm phi) ultrahard zirconium ceramic (Zr) needle and with a common stainless steel (St) needle (also 1.48 mm phi was observed. This was investigated in vivo by histological imaging and biochemical micro-autoradiographic imaging using [2-(14)C]-thymidine as a biomarker in vivo. On the first day after the stab with either, the Zr or the St injection needle, the tunica adventitia showed the most pronounced damage, as evidenced by a large puncture wound characterized by blood congestion, but with few inflammatory cells being observed. A marked contraction of the tunica media was observed. The depth of the injury reached the tunica layer, but amounted to less than 1/3 of the needle diameter. Loose fragments of the endothelial lining were detected, together with scattered red corpuscles. The survival rate of the experimental animals amounted to less than 40% on the 3rd day after the stab by either the Zr or St needle, due to the large needle diameter. In addition, histological imaging of the wound area in the endothelial layer and tunica media showed considerable congestion and inflammation, which limited the evaluation of the regeneration status of the inferior vena cava of the surviving animals. Results were obtained from a few animals that displayed satisfactory recovery status. On the 3rd day after the stab by either the Zr or St injection needle, a relatively large proportion of the hemostatic clots became incorporated into the collagenous tissue, i.e. the tunica adventitia. A marked contraction of the tunica media was also observed, similar to that on the 1st day, following the needle injury. In the case of the endothelium (tunica intima), the injury caused by the Zr needle was reinfiltrated by adult stem cells 3 days after the stab, but the tunica media, composed of endothelial cells, still contained relatively contracted collagenous material. In addition, several interesting cell colonies were observed in the medial layer at the short distance from the boundary of the damaged tissue. It was assumed that these colonies produced medial tissue composed of collagenous supporting tissue or smooth muscle cells. In the experiment using the St needle, the incorporation of [2-(14)C]-thymidine into the nucleus of the stem cells was observed in the small capillaries of the tunica media, but not in the support cells of the latter.

Aspects of inflorescence and floral development in the putative basal angiospermAmborella trichopoda(Amborellaceae)

Amborella has woody axes whose development is intrinsically plagiotropic and determinate. The tree habit is achieved through overtopping of older axes by basally produced younger axes, as in Mangenot's model. Inflorescence units, which are produced in the axils of distal leaves, may be described as extended cymes, each branch ending in a flower. Basal bracteoles have a decussate arrangement, which is modified to an alternate phyllotaxis distally. Flowers produce one or more additional bracteoles with a rapid transition to the spiral phyllotaxis of the broad overlapping tepals. In this transition the initially conical floral apex becomes invaginated to form a floral cup, with subsequent appendages appearing on its inner margin. The floral apex then forms the flat base of the cup but retains a discrete single tunica layer. The receptacular bowl is deepened and narrowed by the basal meristem of each appendage, the last formed floral organs usually consuming the floral meristem. Sexual parts are more numerous in male than female flowers, accounting for their size difference, but primordia of stamens and carpels are initially very similar. Floral symmetry is largely a consequence of close packing of appendages within the floral cup. In its initial stages of development the flower does not conform to any conventional floral model in angiosperms and is better regarded as highly specialized rather than ancestral in its synorganization. This is not unexpected in a lineage of such long independent evolution.Key words: Amborella, basal angiosperm, development, inflorescence, primitive flower.