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Microvascular anatomy of the hippocampal formation.

The hippocampal vessels were examined in 25 forebrain hemispheres injected with india ink or methylmethacrylate. There were two to seven hippocampal arteries, which measured 200-800 microns in diameter. The anterior hippocampal artery (AHA), which was present in 88.2% of the hemispheres, most often originated from the posterior cerebral and the anterior temporal arteries, that is, within the rostral hippocampo-parahippocampal arterial complex. It arose from the anterior choroidal artery in 29.4% of the hemispheres. The AHA extended between the uncus and the parahippocampal gyrus, and it supplied the head of the hippocampus. The middle hippocampal artery was constant. It most often arose from the posterior cerebral and the common temporal arteries. The middle hippocampal artery coursed just caudal to the uncus, in close relationship with the lateral posterior choroidal artery, and it usually supplied the middle part of the hippocampal formation. The posterior hippocampal artery, which existed in 94.1% of the hemispheres, most often arose from the posterior cerebral and the splenial arteries. It irrigated the caudal part of the hippocampal formation. The anastomoses connecting the posterior, middle, and the anterior hippocampal arteries were present in 29.4% of the hemispheres. The hippocampal arteries gave rise to the straight vessels, which divided into the large and the small intrahippocampal arteries. The highest density of the capillary network was noted in the pyramidal and molecular layers of the hippocampal formation. The clinical significance of the obtained microanatomical findings is discussed.

Arteries↗

Experimental omento-myelo-synangiosis.

In 18 cats the omentum was mobilized, transposed, and placed directly upon the pia mater of the spinal cord. Within 2 months, vascular anastomoses developed at the omento-spinal cord interface and, as demonstrated by India ink perfusion, communicated widely with the cord's intramedullary vessels. Light and scanning electron microscopy (SEM) of the interface revealed the development of a "fibrous coat" of Goldsmith in continuity with the edges of the dura mater. It consisted mainly of a matrix of collagen fiber and fibroblasts, well vascularized with sinusoids, capillaries, arterioles and venules. The sinusoids, as identified by SEM, were present in both the fibrous coat and adjacent omentum, were lined with attenuated endothelium, and had multiple communications. The possible significance of the sinusoidal formation is discussed in this report.

Animals↗

The neurovascular relationships and the blood supply of the oculomotor nerve: the microsurgical anatomy of its cisternal segment.

The 27 oculomotor nerves were examined after injection of India ink or methylmethacrylate into the vertebral and the internal carotid arteries. The ventral surface of the cisternal segment of the nerve was noted to be in close relationship with the superior cerebellar artery (96.3%) and its pontine (37.0%), mesencephalic (25.9%) and perforating branches (81.5%); the posterolateral pontine artery (70.4%) and its branches; the anterolateral pontine branches (29.6%), and the perforating branches (85.2%) of the basilar artery; the mesencephalic perforating arteries (11.1%) and their peduncular branches (62.9%); the peduncular branches of the diencephalic perforators (11.1%) and the P1 segment (18.5%); and the accessory collicular artery (3.7%). The dorsal surface of the nerve was in close relationships to the P1 and P2A segments (100%) of the posterior cerebral artery and their peduncular branches (22.2%); the posterior communicating artery (100%); the collicular (100%) and the accessory collicular artery (33.3%), and their peduncular (51.8%) or the perforating branches (22.2%); the medial posterior choroidal artery (25.9%) and its branches (11.1%); and the mesencephalic and diencephalic perforating arteries (100%). Vascular penetration was noted in 51.8% of the third nerves. The most common penetrating vessel was the collicular artery (18.5%) and its branches (22.2%). The cisternal segment of the oculomotor nerve was most often supplied by the mesencephalic perforators (88.9%). The authors discuss the possible clinical significance of the obtained anatomic data.

Humans↗

The anatomical basis for the cerebellar infarcts.

BACKGROUND: Very little can be found in the literature concerning the variation of the irrigation area of the cerebellar arteries, as well as the characteristics of anastomoses among these vessels. The anatomical features may determine certain features of cerebellar infarcts. Consequently, we examined the irrigation area of and the anastomoses among the cerebellar arteries. METHODS: The anatomical features of the posterior inferior cerebellar artery (PICA), the anterior inferior cerebellar artery (AICA), and the superior cerebellar artery (SCA) were studied in 26 cerebella injected with india ink, while their irrigation areas and anastomoses were examined in 8 of these cerebella. RESULTS: The PICA, which most often (82%) arose from the vertebral artery, was found most commonly (81.3%) to supply the largest part of the occipital surface of the cerebellar hemisphere, the caudal or caudomedial part of the tentorial surface, and the inferior vermis. The AICA, which usually (92%) arose from the basilar artery, commonly (68.8%) supplied most of the petrosal surface of the hemisphere and the flocculus. The SCA, which divided into the medial and the lateral trunks, always irrigated most of the tentorial surface of the cerebellum, the superior vermis, and the dentate nucleus. The PICA, AICA, and SCA were always interconnected by anastomoses, which ranged from 40 microns to 420 microns in diameter. CONCLUSIONS: Cerebellar infarcts were documented by computed tomography (CT) or magnetic resonance imaging (MRI) examinations in 10 patients. The infarcts were located in the PICA territory (60%) or the SCA region (40%). The authors compared the obtained anatomic data to the features of the cerebellar infarcts in these patients.

Adult↗

Bladder neck preservation and its impact on positive surgical margins during radical prostatectomy.

To modify the bladder neck dissection during radical prostatectomy, in an effort to improve continence and diminish the incidence of anastomotic stricture, without compromising the primary surgical objective of complete cancer removal. Between December 1991 and August 1992, 50 patients underwent radical retropubic prostatectomy with anatomic dissection and preservation of the bladder neck and most proximal portion of the prostatic urethra, thus creating a mucosal cuff for anastomosis to the urethral stump. There was tumor at the inked margin in 18 patients (36%), however, in only 3 instances (6%) was there tumor at the bladder neck margin. In no instance was the bladder neck margin the only positive margin. At a minimum follow-up of six months, all patients are fully continent during routine activities, and in no patient has an anastomotic stricture developed. Anatomic dissection and preservation of the bladder neck and proximal urethra does not compromise surgical margins. We believe this technique may play a role in preservation of continence after radical prostatectomy and probably decreases the likelihood of anastomotic stricture, by allowing for a circumferential mucosa-to-mucosa anastomosis without the need for bladder neck reconstruction.

Aged↗

A study of prostaglandin F2alpha as the luteolysin in swine: I. Effect of prostaglandin F2alpha in hysterectomized gilts.

Experiments were conducted to determine if prostaglandin F2alpha (PGF2alpha) is luteolytic in swine. In Experiment 1, four bilaterally hysterectomized gilts were injected with PGF2alpha at 0800 (10mg) and 2000 hours (10mg) and four gilts received .9% saline at the same times on day 17 after onset of estrus. Treatments were reversed in the two groups of gilts 21 days later. All eight PGF2alpha treated gilts exhibited estrus an average of 88.0 +/- 13.5 hours after treatment and average duration of estrus was 66.0 +/- 16.4 hours. Saline treated controls did not exhibit estrus. Two additional gilts were hysterectomized bilaterally and the saphenous artery catheterized on day 7 after onset on estrus. PGF2alpha injected on day 17 resulted in a precipitous decline in plasma progestin concentration and onset of estrus by 110 and 90 hours in gilts 1 and 2, respectively. Another bilaterally hysterectomized gilt, with CL marked with India ink, received PGF2alpha on day 17. Estrus occurred 92 hours later and, on day 4, regression of marked CL to corpora albicantia and presence of newly formed CL was confirmed at laparotomy. In Experiment 2, 12 bilaterally hysterectomized gilts were treated with PGF2alpha at 0800 (10mg) and 2000 hours (10mg) on either day 8, 11, 14 or 17 after onset of estrus. None of the gilts treated on days 8 and 11 exhibited estrus. Two of three gilts treated on day 14 and all three gilts treated on day 17 exhibited estrus at an average of 116.0 +/- 9.8 hours post-treatment. Average duration of estrus was 49.6 +/- 8.8 hours.

Animals↗

Effect of indomethacin on estrogen-induced luteolysis in the ewe.

Three groups of 6 ewes were laparotomized on day 9 of an estrous cycle (estrus = day 0) and the corporà lutea (CL) were marked with India ink. Indwelling cannulae were inserted into the uterine horn adjacent to the CL in groups 2 and 3. Group 1 was injected intramuscularly (i.m.) with corn oil twice daily on day 9. Group 2 received 750 microng 17beta-estradiol (E2) i.m. twice daily on day 9 plus intrauterine injections of indomethacin (INDO) vehicle on days 9 through 13. Group 3 received in the same estrogen treatment plus the injection of 20 mg INDO twice daily on days 9 through 13. Jugular venous samples were taken once daily on days 9 through 14 for progesterone analysis. At re-laparotmy on day 14, the ovaries were examined for new ovulations, and the ovary bearing the marked CL was removed. Results showed that E2 induced premature luteal regression as indicated by decreased CL weights and plasma progesterone levels. INDO when given in conjunction with E2 effectively blocked the luteolytic action of E2. These results suggest that the luteolytic action of E2 is mediated via increased prostaglandin secretion and release from the uterus.

Animals↗

Polychrest: a development system for radio-frequency-coupled implants.

Inductively-coupled neurological prosthetic implants may employ RF coils of unusual design (for example 'square' spiral inductors printed in thick-film ink on both sides of the alumina substrate) for which published empirical design equations are hard or impossible to find. 'Polychrest' enables the rapid experimental generation of ad hoc design data for RF-coupled energy-transfer systems, leading quickly to a working model which can be thoroughly studied and understood.

Energy Transfer↗

Meniscal repair by fibrocartilage in the dog: characterization of the repair tissue and the role of vascularity.

Lesions in the avascular part of 20 canine menisci were repaired by implantation of a porous polyurethane. Seven menisci were not repaired and served as controls. The repair tissue was characterized by biochemical and immunological analysis. The role of vascularity in healing was studied by perfusion of menisci with Indian ink. Histologically, repair tissue inside the implants initially consisted of fibrous tissue containing type I collagen. After 2 months, fibrocartilaginous tissue developed inside the implants, whereas control defects only showed repair with fibrous tissue. Both type I and type II collagen, the two major collagen types of normal meniscal fibrocartilage, could be detected in this newly formed fibrocartilage. The implant guided vascular tissue from the periphery towards the lesion resulting in healing of the tear. After fibrocartilage had formed, vascularity decreased and was completely absent in mature fibrocartilage. Control defects remained filled with vascular connective tissue. Two-thirds of the longitudinal lesions were found to be healed partially or completely. It is concluded that implantation of a porous polymer does enhance vascularity sufficiently to result in healing of meniscal lesions extending into the avascular part. Healing takes place by repair tissue strongly resembling normal meniscal fibrocartilage.

Animals↗

Studies on colony specificity in the compound ascidian, Botryllus primigenus Oka. I. Initiation of "nonfusion" reaction with special reference to blood cells infiltration.

When two incompatible colonies of the compound ascidian, Botryllus primigenus Oka come into contact, a specific rejection, called "nonfusion" reaction (NFR), always occurs in the contact area between them. At the beginning of the NFR, concentrations of cells were always observed near the ampullae tips. In the present study, it was revealed that these cells appear to be blood cells infiltrating through the ampullae and were identified as morula cells (vanadocytes). When India ink was injected into vessels prior to the occurrence of NFR, it leaked from the tips of the ampullae into the test matrix, suggesting an increase in the permeability of the ampullar epidermis. Experimental studies to elucidate the mechanism of NFR were carried out. Contact between a colony piece and a zooid-free ampullae fragment or overlapping of one colony to another, respectively, resulted in NFR only at one side of a pair. Furthermore, in stage 3 of NFR, when an opaque area was pushed with a pair of forceps and the blood in this area was transferred to healthy part of vessels, the stoppage of blood flow and subsequent contraction of vessels always took place in the latter. These observations suggest that ampullae penetration into the opposite colony is essential to NFR and that NFR is directly initiated in the ampullar lumen.

Animals↗

Method for the determination of cerebral blood flow in conscious rabbits.

A procedure for the determination of cerebral blood flow by the local clearance method after intracarotid injection of 133Xe in the conscious rabbit is described. The inert radioactive indicator is injected into a permanent nylon catheter equipped with a two-way Gordth's needle inserted into the common carotid artery and filled with heparin, emerging behind the shoulders of the animals. All branches of the homolateral common carotid artery except the internal carotid artery were ligated. Studies of the distribution of colored tracers (dark blue ink) and radioactive tracers (99mTc albumin microspheres) show that the main localization of the injected indicator is within the homolateral hemisphere. Brain to blood partition coefficients of 133Xe are worked out for rabbit's gray matter (0.576 +/- 0.048) and white matter (0.808 +/- 0.023). The slope method for first and second component of the wash-out Xenon curve is used for CBF calculations. CBF determinations in 9 normal rabbits result in 84.27 +/- 5.59 and 16.69 +/- 2.44 ml/min x 100 g tissue, respectively, for the fast and slow component. Significant changes do not occur in serial determinations within 2 hr.

Anesthesia↗

Protein blotting on nitrocellulose: some important aspects of the resolution and detection of antigens in complex extracts.

The resolution and detection of individual components in complex extracts by protein blotting have been investigated. By probing nitrocellulose transfers with monospecific and multispecific antisera, it was demonstrated that dissociating conditions were required for the maximum resolution of antigens by polyacrylamide gel electrophoresis, a conclusion reinforced by results from 2-D electrophoresis. The dissociating and reducing treatments employed, however, were both shown to be responsible for some loss of total antigenicity and included the complete loss of at least one important antigen. Assays with nitrocelluloses of different pore sizes demonstrated that both higher protein-binding capacities and higher backgrounds were associated with the use of the smallest pore size, while the sensitivity of the assay was greatest when a non-ionic detergent, and not proteins, were used for blocking. Nitrocellulose-bound proteins may be stained with amido black, India ink, toluidine blue, Ponceau S or a gold sol, but these agents do not always give identical staining patterns. While detection of components with immuno-enzyme staining methods had some advantages, problems with non-specific binding were encountered. These did not occur with affinity purified radiolabelled second antibodies, which in combination with scanning of autoradiographs allowed a quantitative approach to be adopted.

Animals↗

Spectrophotometric determination of effective proteolytic activity in biodetergents.

A new procedure for the determination of effective proteolytic activity in biodetergents has been developed. Effective enzyme activity is defined as the activity exhibited by the tested enzyme under real washing conditions, i.e., in the water suspensions of the biodetergent tested at the working temperature (usually 40 degrees C). Two insoluble chromolytic substrates, namely black gelatin (gelatin cross-linked with glutaraldehyde in the presence of black drawing ink) and the protease substrate based on the immobilization of dyed casein in the structure of polyacrylamide gel (blue casein-PAAG) were successfully used. It was found that there is a great difference in effective proteolytic activity among various biodetergents. The proposed procedure is suitable both for the manufacturers for a quick control of their products and for the quality control laboratories.

Detergents↗

Reflectance of rat brain structures mapped by an optical fiber technique.

A method to detect differences in reflectance as a pair of glass fibers is advanced through various brain structures at a constant speed is described. The system has been used to develop a technique for accurate and easy placement of electrodes or cannulae into a limited brain region. The probe consisted of two thin (50 micrometer in diameter) glass optic fibers. A 5 W lamp was used to transmit light down one of these fibers. The relative intensity of light reflected from brain tissue into the other optic fiber was monitored with a photomultiplier and ink-writing recorder. Different brain structures were shown to vary in the amount of reflectance, with white matter having much higher reflectance than gray matter. It was demonstrated in a total of 239 penetrations that the patterns of successive changes in the reflected light response, as the probe was lowered into the brain, was characteristic for each frontal plane. The probe may additionally be itself used as an electrode if plated with silver.

Animals↗

Comprehensive polypeptide analysis of microdissected rat brain areas: combining 2-dimensional gel electrophoresis with 2-dimensional HPLC and immunoanalysis and sequencing procedures.

An improved methodology has been developed which allows resolution, identification, and quantitation of hundreds of proteins and neuropeptides from a single rat brain nucleus (5 mg wet wt.). After metabolic labelling, proteins (greater than about 15 kDa) are separated from peptides (less than about 10 kDa) by sonicating the tissue in an acidic peptide extraction buffer; after centrifugation, proteins are in the pellet, peptides in the supernatant. To quantitate peptide synthesis, peptides are resolved to purity by reverse-phase HPLC followed by ion exchange HPLC. Proteins are resolved with a two-dimensional (2-D) gel protocol optimized for neural tissue. To identify specific proteins by immunoanalysis, proteins are transferred to polyvinyl difluoride (Immobilon) and immunostained in the presence of Tween blocking buffer. After visualization with an avidin-biotin alkaline phosphatase procedure, the blot is post-stained with India ink to visualize the protein pattern context. To sequence spots, proteins are transferred to Immobilon, stained with Coomassie, and directly subjected to automated gas phase sequencing. The immunoblot procedure can detect less than 0.1 pmol protein, and the sequencing procedure can detect less than 10 pmol protein. After transfer enough material remains on the gels to allow subsequent autoradiography or silver stain. Quantitative analysis of 2-D gels is examined in a companion paper. These procedures should enhance the utility of 2-D gels in neurochemical studies.

Amino Acid Sequence↗

Studies on three structurally related phenylenediamines with the mouse micronucleus assay system.

Three structurally related compounds, 4-chloro-o-phenylenediamine (COP), 4-nitro-o-phenylenediamine (NOP) and p-phenylenediamine dihydrochloride (PPD), are used in fur dyes, inks and hair coloring formulations. COP has been reported to be carcinogenic in both rats and mice. NOP and PPD are non-carcinogens, but have consistently tested positive in short-term in vitro genotoxicity assays. Studies were undertaken to evaluate their genotoxicity with the in vivo mouse bone-marrow micronucleus assay. Five CD-1 male mice per dose were injected i.p. with the compounds and sacrificed at intervals of 24, 48 and 72 h. 2000 cells were scored per animal to determine the frequency of micronucleated-polychromatic erythrocytes (MPCE). COP induced significant dose-related increases in MPCE over the 3 doses tested at each of the sampling intervals. The peak response occurred at 24 h. No response was observed in animals treated with PPD or NOP.

Animals↗

Application of an indirect carbon immunoassay (CIA) for the rapid diagnosis of antibody to Toxoplasma gondii in sheep.

Carbon immunoassay (CIA), a novel indirect rapid test for Toxoplasma antibody in sheep, was compared with indirect fluorescent antibody assay (IFA). CIA relies on the adherence of carbon particles of India-ink to rabbit immunoglobulin G.l Carbon labelled anti-sheep rabbit IgG was used for the detection of sheep antibody when attached to tachyzoites of Toxoplasma gondii. The result was read in an ordinary light microscope and there was a clearcut difference between negative and positive reactions. Out of a total of 97 sheep sera tested, 15 sera were negative in both tests and 12 were negative in CIA but showed low positive titres in IFA. The remaining 70 sera were positive in both tests but the titres were usually about 3 dilution steps lower when investigated with CIA as compared to IFA.

Animals↗

Structure and function of the melano-macrophage centres of the goldfish Carassius auratus.

We have studied the melano-macrophage centres (MMCs) of Carassius auratus as well as the changes they undergo in different experimental conditions. The MMCs of the spleen contain brown and yellow pigments, but dark pigments abound in those of the kidney. They occur in association with splenic ellipsoids and scattered throughout the renal parenchyma. Histochemical tests demonstrated small amounts of lipids and basic proteins, abundant neutral mucopolisaccharides and differences between the spleen MMCs and those of the kidney, specially regarding the presence or absence of haemosiderins and melanins. Electron microscopy shows them to consist of phagocytes containing degenerated debris, sometimes surrounded by a discontinuous layer of flat reticular cells. The MMCs of the spleen and kidney trap carbon material 48 hours after injection of Indian ink. Primary and -more so- secondary immunization with SRBC produces a net increase in number and size of MMCs. Treatment with phenylhydrazine produces a notable and fast increase of the MMCs in both organs. After 5 days the MMCs degenerated and erythroblasts restored the erythroid population. Apparently, MMCs represent non-specific means of phagocytosis related to different physiological processes.

Animals↗