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Absence of postnatal death among motoneurones supplying the inferior gluteal nerve of the rat.

Motoneurones innervating the caudal part of the gluteus maximus muscle of 0-2 day, 10-12 day and 2-3-month-old rats were labelled by a half-hour application of a solution of 30% horseradish peroxidase (HRP) and 2% lysophatidylcholine delivered by suction electrode to the cut inferior gluteal nerves. The numbers of motoneurones labelled 24-48 h later were not significantly different in the 3 age groups (mean = 58.75, 54.0, 57.5, respectively). When a simple 30% HRP solution was used in adult rats, the number of motoneurones labelled was significantly less (mean = 48.75). In contrast, application of 0.5 microliter of HRP in a pledget of gelfoam to either the cut or uncut inferior gluteal nerve of neonates labelled large numbers of motoneurones, presumably by diffusion into nearby muscles. It is concluded that no death of motoneurones innervating the gluteus maximus muscle occurs postnatally, and that spread of HRP to neighbouring muscles can give rise to spuriously high motoneurone counts in neonates, and that incomplete uptake or transport of HRP in adults can lead to incorrectly low counts.

Age Factors↗

Karyotype evolution in a simian virus 40-transformed tumorigenic human cell line.

Normal human foreskin fibroblasts (HSF4) were transfected using the pSV3-neo plasmid. A pool of 10 G418-resistant colonies, HSF4-T12, showed a progressive increase in the expression of a number of in vitro transformation markers with passage in culture and became immortalized. Although no tumors were formed when cells were injected subcutaneously into nude mice, this cell line produced progressive tumors when cells were injected into preimplanted Gelfoam sponges in the mice. When these tumors were cultured in vitro and subsequently injected subcutaneously, progressive tumors were produced with median latency periods as short as 4 weeks. Three phases of cytogenetic change could be distinguished. At early passages after transfection. HSF4-T12 exhibited many random chromosomal changes. At a time just after immortalization, both flow karyotype and G-banded analyses showed the appearance of balanced clonal rearrangements. These included t(2;4), t(2;14), t(3;?), 6p-, i(6p), 8p-, t(14;15), i(15), and t(18;?). These clonal rearrangements were stable with passage in culture, and less variability from cell to cell was noted. The only consistent chromosomal loss observed was -Y. Analysis of three independent tumors showed characteristic loss of chromosomal material rather than balanced chromosomal rearrangements. Frequent loss of 6q and chromosomes #13, 15, 20, and Y was noted.

Animals↗

Structure of the pars flaccida after occlusion of the Eustachian tube or blockade of the tympanic isthmus.

In this experimental study performed on the rat middle ear either the Eustachian tube or the tympanic isthmus was blocked via the tympanic bulla with a polyethylene plug or a piece of Gelfoam, respectively. Effusion material was immediately observed in the attic space and subsequently the pars flaccida was drawn in a medial direction to form a retraction pocket. The initial changes in the pars flaccida structure were a degranulation of the mast cells and concomitantly metaplasia of the mucosal epithelium into cells displaying numerous osmiophilic inclusions, vacuoles and multivesicular bodies. It cannot be excluded that the initial cause of the effusion production is the histamine released from mast cell granules.

Animals↗

A randomized trial of hepatic arterial chemoembolization in patients with unresectable hepatocellular carcinoma.

A randomized trial of hepatic arterial chemoembolization was conducted in 42 patients with unresectable hepatocellular carcinoma. These patients represented 41% of patients with hepatocellular carcinoma seen during the inclusion period. In the remaining 59%, 9% had resectable tumours and 50% had unresectable tumours with contraindication for chemoembolization. Patients received either repeated chemoembolization with gelfoam powder and doxorubicin (group 1) or symptomatic treatment (group 2). There was no difference in age, prevalence of cirrhosis or staging according to Okuda between the two groups of patients. A complete tumour response (assessed by arteriography, ultrasonography and serum alphafetoprotein) was observed in four patients, and a partial response in three other patients from group 1. Actuarial survival rates were 33 and 24% in group 1 and 52 and 31% in group 2 at 6 and 12 months, respectively (differences were not significant--logrank test). With the treatment used in our study, chemoembolization did not prolong the survival time of patients with unresectable hepatocellular carcinoma. There were, however, some complete or partial responses. The high spontaneous 1-year survival rate of untreated patients was probably due to the exclusion of the most severely ill patients. Our results do not support the use of this method of chemoembolization in the treatment of hepatocellular carcinoma.

Adult↗

Messenger RNA and protein expression of basic fibroblast growth factor receptor after cortical ablation.

In a previous report we demonstrated that basic fibroblast growth factor (bFGF), as a multipotent neurotrophic factor, could prevent retrograde degeneration of the thalamic neurons after ablation of the somatosensory cortex. To elucidate the mechanism of this bFGF action, we examined changes in FGF receptor (FGFR) mRNA (flg) expression with in situ hybridization. The FGF receptor protein was detected with the immunoblotting method. The FGFR mRNA expression was found to be diffusely increased in the affected cortex. Microscopic observation indicated that FGFR mRNA was expressed in several types of cortical cells including neurons and non-neuronal cells. This increase could be observed as early as 6 hours after surgery and lasted for 48 hours. In the thalamus, however no change in FGFR mRNA signals was observed. Western blotting detected a protein immunoreactive to anti-FGFR antibody. Samples from the periablated cortex showed an increase in FGFR protein. Samples from the thalamus, however, showed no difference in FGFR protein level between the lesion side and the contralateral side. Application of exogenous bFGF in Gelfoam to the cortical ablation cavity did not show any effect on the gene expression or protein level of FGFR. These results suggest that FGFR is diffusely induced throughout the injured cortex in the early phase after injury and that bFGF may play an important role after injury. Topically applied bFGF might thus modulate cellular responses in the cortex and have a neurotrophic effect on the affected thalamic neurons.

Animals↗

Ciliary neurotrophic factor supports target-deprived preganglionic sympathetic spinal cord neurons.

Spinal cord neurons in the intermediolateral column (IML) that innervate the adrenal medulla require target-derived factor(s) for their maintenance in vivo. Selective destruction of the adult rat adrenal medulla causes an approximate 25% loss of Nissl-stained IML neurons between spinal cord levels Th 4 weeks after surgery. We have previously suggested that basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF) or closely related molecules are present in adrenal chromaffin cells and granules. Basic FGF supplemented in gelfoams to the medullectomized adrenal gland fully prevented IML neuron losses. These results are now extended by demonstrating that (i) CNTF administered in vivo (7.2 micrograms/gelfoam) also rescues IML neurons, and (ii) the rescue effect is abolished by splanchnicotomy, i.e. interruption of axonal pathway from the spinal cord to the adrenal gland. These data would be consistent with CNTF being a retrograde trophic messenger to the spinal cord, but do not exclude the possibility that CNTF mimics or induces the formation of an endogenous trophic factor.

Adrenalectomy↗

Time course of the neuroprotective effect of transplantation on quinolinic acid-induced lesions of the striatum.

Injection of quinolinic acid in the rat striatum mimics neurochemical changes observed in Huntington's disease. We previously demonstrated that intrastriatal transplantation of fetal striatum or gelfoam protects against toxicity induced by a subsequent intrastriatal injection of quinolinic acid performed one week later. Herein, we examined whether fetal striatum or sham transplantation provides protection against quinolinic acid that lasts up to four weeks. Intrastriatal quinolinic acid injection produces neuronal loss and gliosis in Nissl staining, loss of cytochrome oxidase histochemical staining, decrease in autoradiographic binding of [3H]SCH 23390-labeled dopamine D1 and [3H]CGS 21680-labeled adenosine A2 receptors, and increase in autoradiographic binding of [3H]PK 11195-labeled peripheral benzodiazepine binding sites. None of these changes was observed in rats transplanted with fetal striatum one, two or four weeks before quinolinic acid injection. In animals transplanted with fetal striatal tissue, Nissl staining showed healthy grafts located in normal appearing striata. Although sham transplantation performed one week before quinolinic acid injection also protected against histological, histochemical and binding changes, sham transplantation performed two or four weeks before quinolinic acid injection was less effective in attenuating quinolinic acid-induced striatal toxicity. Thus, sham transplantation provides transient protection against quinolinic acid-induced striatal toxicity, whereas implantation of tissue such as fetal striatum seems to be required for long-lasting protection. Our study suggests that intracerebral transplantation may also act through other mechanisms than restoration of deficient neurotransmitters or damaged pathways, a finding which may have significant clinical implications in assessing the potential benefit of this approach for the treatment of neurodegenerative disorders such as Huntington's disease.

Adenosine↗

AChE-positive fiber growth after hippocampal fimbria transection and peripheral nerve homogenate implantation.

Gelfoam treated with peripheral nerve homogenate was implanted into a site of hippocampal fimbria transection in the adult rat to assess whether the homogenate might enhance growth of AChE-positive fibers into the lesion site and whether the fiber growth might be directed to the implants. Homogenate was prepared from intact peripheral nerve and in a few cases from degenerated nerve. Some implants were encased in Silastic. Homogenate was also injected into the denervated hippocampus. The major finding was that AChE-positive fiber growth was associated with regions of high astroglial cell content in preference to the relatively hypocellular implants. No clear differences in AChE fiber sprouting into the lesion site, fiber growth into implants, or hippocampal reinnervation were noted between homogenate and saline-treated animals.

Acetylcholinesterase↗

Transport of molecules from nose to brain: transneuronal anterograde and retrograde labeling in the rat olfactory system by wheat germ agglutinin-horseradish peroxidase applied to the nasal epithelium.

Transneuronal anterograde labeling with the conjugate wheat germ agglutinin-horseradish peroxidase (WGA-HRP) has been documented in the mammalian and immature avian visual system [6,14]. Transneuronal retrograde labeling was significant only in the chick [6]. The present study was performed to determine whether transneuronal labeling could be shown in the mammalian olfactory system, whether the phenomenon was robust in adults, and whether transneuronal retrograde transport could label several transmitter-specific centrifugal afferent projections to the olfactory bulb. In addition we wished to learn whether molecules that enter the nasal cavity can undergo transport to brain neurons. Gelfoam implants soaked with 1% WGA-HRP, surgically implanted into the nasal cavity, produced transneuronal labeling patterns that affirmed all of these questions. Transneuronal anterograde transport labeled the appropriate zones in the olfactory bulb and in all second order olfactory targets. In addition, there was transneuronal retrograde labeling of neurons in the olfactory bulb, anterior olfactory nucleus and in transmitter-specific projection neurons from the diagonal band (cholinergic), raphe (serotonergic) and locus coeruleus (noradrenergic). Transneuronal labeling was robust and consistent. The patterns of labeling indicated that transneuronal anterograde and retrograde transport occurred along known, specific circuits in the olfactory system. The present results suggest that nasal epithelial application of WGA-HRP may be a useful tool for assessing regeneration of primary olfactory neurons and the status of central circuitry following regeneration. The method should also facilitate the study of central olfactory connections after surgical or genetic lesions of the olfactory bulb. Finally, these experiments suggest the possibility that inhaled molecules including, possibly substances of abuse, may be transported to, and, possibly, influence the function of neurons in the brain, including some (diagonal band, raphe, locus coeruleus) which have extensive projections to wide areas of the CNS.

Animals↗

Histological evidence for hair cell regeneration after ototoxic cell destruction with local application of gentamicin in the chinchilla crista ampullaris.

Two experiments were conducted to study the ototoxic effects of local gentamicin (GM) administration and the subsequent hair cell (HC) regeneration process in the chinchilla cristae ampullares (CA). In the first experiment, 3 different doses of GM (0.1, 0.2 and 1.2 mg) were administered by surgical implantation of GM-soaked Gelfoam pledgets in the perilymphatic space in the otic capsule of the left superior semicircular canal. The CA was histologically processed for light-microscopic examination. In the second experiment, 6 groups of 2 chinchillas each were treated with 0.1 mg of GM. To document cell proliferation and HC regeneration, Alzet micro-osmotic pumps were implanted in each chinchilla to deliver bromodeoxyuridine (BrdU) at 125 micrograms/h for 1 week. Chinchillas were subsequently killed at 1 and 4 days and 1, 2, 4 and 8 weeks post-treatment (PT). The CA was processed for light microscopy and BrdU immunocytochemistry. In the first experiment the smallest dose produced damage restricted to HCs alone, while the medium and large doses produced severe damage in the sensory epithelium, including supporting cells and HCs. Results in the second experiment demonstrated that at 1 and 4 days PT the HCs showed extensive damage, including clumping of nuclear material. By 4 days PT the supporting cell nuclei lost their monolayer configuration. Calyceal terminals appeared empty, and vacuolized remnants of nerve calyces were evident in the basal portion. At 1 week PT complete disappearance of HCs from the sensory epithelium was evident, and there was cytoplasmic extrusion into the endolymphatic space. At 2 weeks PT there was complete HC loss, the supporting cell nuclei were scattered randomly in the crista, and the nerve fibers were retracted from the sensory epithelium. At 4 weeks PT there was evidence of sensory epithelium repair and HC regeneration. Short cells resembling type-II HCs were evident in the surface of the sensory epithelium. At 8 weeks PT the number of HCs increased in a uniform fashion on the surface of the sensory epithelium, and the supporting cell nuclei were realigned on the basal membrane. Nerve fibers with growth cones penetrated the basal membrane. Supporting cell proliferation was evident by the presence of mitotic figures and BrdU immunoreactivity in the chromatin material of dividing cells at 2 weeks PT. The labeling was more evident in newly formed cells at 4 and 8 weeks PT. These results demonstrate that in chinchillas the vestibular organs have the capacity of self-repair and the process includes HC regeneration after local administration of GM. The overall process involves changes in different cells in the sensory epithelium and neural elements, all of which show modifications with an orderly pattern.

Animals↗

Splenic embolization prior to myelosuppressive treatment in hepatocarcinoma and active chronic hepatitis.

In the treatment of active chronic hepatitis and hepatocellular carcinoma some effective drugs can produce myelosuppression. Hypersplenism may considerably limit the dosage of such drugs. Splenectomy is an effective treatment for hypersplenism, although it is not without complications. Partial splenic embolization is a good and safe procedure; 15 patients were treated in order to achieve higher platelet and leukocyte counts. Embolization has been performed with gelfoam with local and systemic antibiotics (Spigos' protocol) and 50-75% of the splenic parenchyma was infarcted. All patients could be treated for the underlying hepatopathy with adequate dosages of interferon or chemotherapeutic drugs.

Aged↗

The MR appearance of endovascular embolic agents in vitro with clinical correlation.

A spectrum of embolic agents including PVA, avitene, gelfoam, ethanol, IBCA, silicone microspheres, metal and platinum coils, and balloons containing contrast of HEMA were imaged in vitro at 0.3 Tesla using spin echo, gradient echo, and inversion recovery sequences. Signal intensities associated with these agents and changes created by addition of blood are presented. The in vitro model is also correlated with clinical MR material. Recognition of embolic materials is important to avoid confusion with persistent flow void, hemorrhage, or thrombus.

Adult↗

Endothelin-mediated induction of heme oxygenase-1 in the spinal cord is attenuated in transgenic mice overexpressing superoxide dismutase.

Spinal cord blood flow and the induction of heme oxygenase-1 (HO-1), an indicator of oxidative stress, were studied in the spinal cords of adult wild-type and transgenic mice overexpressing the antioxidant copper, zinc superoxide dismutase (CuZn SOD) after intrathecal administration of the potent vasoactive peptide endothelin-1 (ET-1). Gelfoam, saturated with ET-1 (40, 80, or 400 micromol/L), was positioned in the intrathecal space at the midthoracic level in anesthetized animals. Blood flow was continuously monitored by laser Doppler for 10 min after the intrathecal application of ET-1. There was a significant reduction in spinal cord blood flow to approximately 40% of control values by 10 min after the intrathecal application of the peptide in both wild-type and transgenic mice. Moreover, SB209670, a nonselective endothelin receptor antagonist, blocked this reduction in flow. Each animal was euthanized 24 h after the intrathecal administration of ET-1, and the spinal cord was prepared for quantitative immunocytochemistry. HO-1 was primarily induced in astrocytes near the dorsal surface of the spinal cord in wild-type mice. This induction was attenuated in both wild-type, treated with SB209670, and untreated transgenic mice. Together, these findings suggest that ET-1 mediates oxidative stress in the spinal cord through the modulation of spinal cord blood flow.

Animals↗

Tumour seeding in peritoneal wound sites in relation to growth-factor expression in early granulation tissue.

The purpose of these experiments was to identify growth factors produced during the formation of a peritoneal wound in relation to tumour cell seeding and stimulated growth in granulation tissue. Gelfoam(c) gelatin sponge was implanted in the mesenteric fan of nude mice to initiate the granulation process. Human HT29 colon carcinoma cells were inoculated intraperitoneally at various times after sponge implantation and tumour growth in granulation tissue was determined. RNA isolated from granulation tissue was used for polymerase chain reaction analysis of the expression of specific growth factors and receptors [vascular endothelial growth factor (VEGF), transforming growth factor-beta (TGF-beta) and lysophosphatic acid (LPA)], and for microarray analysis of differentially expressed genes in early vs. late granulation tissue. Inflammatory cells infiltrated the sponge within 1 day, followed by fibroblasts and the formation of an extracellular matrix. Tumour cell inoculation at 8 h to 3 days after sponge implantation resulted in extensive tumour formation in all cases. Inoculation at 10-28 days resulted in focal tumour growth in only 16-33% of the sponges. Low amounts of VEGF, TGF-beta(1-3), TGF-beta RIII and LPA receptors 1,2 were detected in early granulation tissue, with increased expression from day 10. Microarray analysis identified additional differentially expressed genes that may stimulate tumour take and growth in early granulation tissue.

Animals↗

Edaravone protects the vestibular periphery from free radical-induced toxicity in response to perilymphatic application of (+/-)-alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid.

Intracochlear infusion of (+/-)-alpha-amino-3-hydroxy-5-methyl-isoxazole-4-propionic acid (AMPA) was performed with a syringe pump in guinea pigs, and peripheral vestibular dysfunction was induced. Animals were administered edaravone systemically or topically. In the systemic application group, animals were administered edaravone once a day for 7 days after AMPA infusion. In the topical application group, edaravone-soaked gelfoam was placed on the round window membrane just after, 12 h after or 24 h after AMPA infusion. Spontaneous nystagmus was observed after AMPA infusion. Immunohistochemistry for 4-hydroxy-2-nonenal (4-HNE), a marker of free radical-induced lipid peroxidation, was performed 24 h after AMPA infusion. In addition, caloric tests were performed to evaluate vestibular function 1 week after AMPA infusion. Animals in both groups showed decreased spontaneous nystagmus, but results were not significant. Animals treated topically with edaravone within 12 h of AMPA infusion showed normal morphology of the ampullar sensory epithelia of the lateral semicircular canals and showed a good response to the caloric tests. 4-HNE immunoreactivity in the sensory epithelia was very low in these animals. In contrast, untreated animals and animals treated with edaravone systemically or topically 24 h after AMPA infusion showed morphologic hair cell damage, reduced caloric response and remarkable 4-HNE immunoreactivity in the sensory epithelia. These results indicate that topical application of edaravone within 12 h after damage protects the vestibular periphery from free radical-induced toxicity in response to intracochlear infusion of AMPA.

Aldehydes↗

Percutaneous uterine artery embolization for the treatment of symptomatic fibroids: current status.

Uterine artery embolization (UAE) is increasingly being used as an alternative treatment to hysterectomy for symptomatic fibroids. Symptoms of pelvic pressure, urinary frequency and menorrhagia are controlled in 73-98% of patients who undergo UAE. At the 1-year follow-up, the uterus may shrink by up to 55% but re-growth of fibroid may however occur. The rate of major complications and amenorrhoea following this procedure is low, ranging in most series from 1 to 3.5% and 1 to 7%, respectively. Nevertheless, the rate of amenorrhoea in women over 45 seems to be higher. In order to completely block the arterial supply to the fibroid, UAE is typically performed in both uterine arteries. Different embolic agents are used such as polyvinyl alcohol, gelfoam and more recently gelatine tris-acryl microspheres. After UAE, perfusion of the uterus is maintained. Uterine function is therefore conserved and although women who become pregnant after UAE seem to be at risk for malpresentation, pre-term birth, cesarean delivery and postpartum hemorrhage, successful pregnancies after UAE have been reported in some series. A major technical problem with UAE remains the possible presence of fibroid blood supply from other sources, such as the ovarian arteries or other pelvic branches, which can lead to failure of the procedure. In conclusion, although randomized trials are still underway, UAE appears a good option for those patients who whish to conserve their fertility or when surgery is contra-indicated. However, to evaluate the long-term effects of UAE longer follow up is required.

Diagnostic Imaging↗

Protection of auditory function against noise trauma with local caroverine administration in guinea pigs.

Glutamate is the most likely neurotransmitter at the synapse between the inner hair cell and its afferent neuron in the peripheral auditory system. Intense noise exposure may result in excessive glutamate release, binding to the post-synaptic receptors and leading to neuronal degeneration and hearing impairment. The present study investigated the protective effect of caroverine, an antagonist of two glutamate receptors, N-methyl-D-aspartate and alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid, on noise-induced hearing loss. Two different doses of caroverine were applied onto the round window membrane with gelfoam, followed by one-third-octave band noise centered at 6.3 kHz (110 dB SPL) for 1 h. Auditory brainstem responses were measured at regular time intervals afterwards. Caroverine was found to offer significant protection of the cochlear function against noise-induced hearing loss.

Animals↗

Depressed fractures over cranial venous sinus.

Seventeen patients with depressed fractures over cranial venous sinus who had undergone operations between 1993 and 1998 were reviewed retrospectively. There were 15 male and two female patients. Their ages ranged from 7 to 48 years. The most frequent causes of trauma were motor vehicle accident (in eight cases) followed by assault (in six cases). In the majority of cases, depressed fractures were localized over the superior sagittal sinus, and half of them were over the medial third of the sinus. In 11 cases, massive blood loss occurred intraoperatively that could be controlled by digital pressure with gelfoam or with a free muscle flap. Postoperative infection developed in one case and was treated successfully with antibiotic medication alone. Two patients died due to associated intracerebral lesions. The remaining 15 patients returned to daily living activity in the follow-up period.

Adolescent↗