Intra-arterial chemotherapy is reliable in preventing high-risk superficial bladder cancer from recurrence and progression.

The purpose of this prospective study was to evaluate the therapeutic effects of intra-arterial chemotherapy in preventing high-risk superficial bladder cancer from recurrence and progression. from may 2003 to December 2007, 52 patients were divided randomly into 2 groups. Twenty-five patients were given intra-arterial chemotherapy with gemcitabine and cisplatin, and 27 patients received intravesical instillation with epirubicin. After 6-67 months of follow-up (median, 40 months), the overall recurrence-free rates of the intra-arterial chemotherapy and intravesical instillation groups were 83.3% and 33.4%, respectively (p=0.001 log rank). Tumor progression was not found in the intra-arterial chemotherapy group while 7 patients in the intravesical instillation group had tumor progression. The overall tumor progression-free rates were 100% and 58.5%, respectively (p=0.009 log rank). The patients with functional bladders were 100% and 81.5% in the intra-arterial chemotherapy and intravesical instillation groups after 67 months of follow-up, respectively. In conclusion, intra-arterial chemotherapy is more effective than intravesical instillation in preventing high-risk superficial bladder cancer from recurrence and progression.

Incidence of complications after temperature-controlled radiofrequency treatment for sleep-disordered breathing: a Singapore sleep centre experience.

OBJECTIVE: To determine the incidence of complications following temperature-controlled radiofrequency treatment of the soft palate, uvula and tongue base. STUDY DESIGN: Retrospective study. SETTINGS AND METHODS: We included all patients who had received temperature-controlled radiofrequency treatment of the soft palate, uvula and tongue base, for sleep-disordered breathing, over a four-year period in a tertiary hospital. Patients' medical records were systematically reviewed for radiofrequency treatment parameters and complications. MAIN OUTCOME MEASURE: Complication rates. RESULTS: Seventy-six patients had been treated, with a total of 127 treatment sessions and 544 lesions to the palate, uvula and tongue base. The incidences of minor and moderate complications were, respectively, 2.6 per cent (14/544 lesions) and 0.4 per cent (2/544 treatment lesions), being 3.0 per cent (16/544 lesions) overall. Subdividing by anatomical region, the incidences of minor and moderate complications following palatal and uvula radiofrequency treatment were, respectively, 3.1 per cent (14/446 lesions) and 0 per cent, and those following tongue base treatment were, respectively, 0 per cent and 2.0 per cent (2/98 lesions). The incidence of minor complications following soft palate and uvula treatment, per treatment session, was 10.9 per cent. The incidence of moderate complications following tongue base treatment, per treatment session, was 4.6 per cent. There were no major complications in our study population. CONCLUSIONS: In this study, the incidence of complications of temperature-controlled radiofrequency treatment of the palate, uvula and tongue base was low. Temperature-controlled radiofrequency is a safe treatment modality for patients with sleep-disordered breathing and can be performed as a day case procedure. We recommend day admission for patients undergoing radiofrequency of the tongue base, in view of the potential for severe complications and airway compromise.

Migrating esophageal foreign bodies.

UNLABELLED: Ingested foreign bodies which migrate extraluminally, although rare in occurrence, are fraught with the potential to cause life-threatening complications. PURPOSE OF THE STUDY: To discuss the management of this pathology. MATERIAL AND METHODS: A series of four patients with such occurrences is presented. CONCLUSION: A discussion on the safe management of such seemingly innocuous foreign bodies allows the authors to propose a therapeutical algorythm.

Using biosensors to detect the release of serotonin from taste buds during taste stimulation.

CHO cells transfected with high-affinity 5HT receptors were used to detect and identify the release of serotonin from taste buds. Taste cells release 5HT when depolarized or when stimulated with bitter, sweet, or sour tastants. Sour- and depolarization-evoked release of 5HT from taste buds is triggered by Ca2+ influx from the extracellular fluid. In contrast, bitter- and sweet-evoked release of 5HT is triggered by Ca2+ derived from intracellular stores.

The distribution of PGP9. 5, BDNF and NGF in the vallate papilla of adult and developing mice.

The development and innervation of vallate papillae and taste buds in mice were studied using antibodies against the neuronal marker, protein gene product 9.5 (PGP 9.5), and against nerve growth factor (NGF) and brain-derived neurotrophic factor (BDNF). PGP 9.5 immunohistochemical studies revealed that the earliest sign of median vallate papilla formation was an epithelial bulge at embryonic day 13 (E13), and at E14, a dense nerve plexus was found within the connective tissue core of the papilla. Thin nerve fibers penetrated the apical and medial trench wall epithelium of the papilla at E16 and a few of these began to invade the lateral trench wall epithelium at E17. At postnatal day 1 (P1), the newly formed taste buds were recognizable and a small number of PGP 9.5-immunoreactive (IR) cells appeared on the medial trench wall epithelium. The number of PGP 9.5-IR taste bud cells then increased gradually and reached the adult level at postnatal week 2. PGP 9.5 immunoreactivity increased systematically with age. NGF and BDNF immunoreactivity was first seen at the boundary between the columnar cells in the apical epithelium of the developing vallate papilla at E13, then in the medial and lateral trench walls at E15 (BDNF) or E18 (NGF). At P1, BDNF immunoreactivity was exclusively present in the newly formed taste buds of the medial trench wall. The number of BDNF-IR taste bud cells then increased gradually, reaching the adult level at P7. Similar degrees of NGF and BDNF immunoreactivity were seen in the developing vallate papilla. In the present study, we found that the vallate papilla was formed prior to its innervation, and we propose that initiation of papilla formation does not require any direct influence from the specific gustatory nerve. We also suggest that neurotrophins in the early developing vallate papillae might act as local tropic factors for the embryonic growth of nerve fibers to induce differentiation of the taste buds.

Effects of zinc deficiency on the vallate papillae and taste buds in rats.

BACKGROUND AND PURPOSE: Zinc deficiency is associated with multiple clinical complications, including taste disturbance, anorexia, growth retardation, skin changes, and hypogonadism. We investigated the zinc-deficiency-induced morphologic changes in the vallate taste buds of weanling and young adult male Wistar rats. METHODS: A total of 24 weanling and 30 young adult rats were used. Each age group was further divided into a control group fed a zinc-adequate (50 ppm) diet, a zinc-deficient (< 1 ppm) diet group, and a zinc-adequate pair-fed group who were fed the same amount of food as that taken by the zinc-deficient group. Weanling rats were fed for 4 weeks and young adult rats were fed for 6 weeks. The morphometry and morphologic changes of vallate taste buds were analyzed using light and transmission electron microscopy. RESULTS: Light microscopy revealed no significant difference in papilla size and morphology among the various groups. In both weanling and young adult rats in the zinc-deficient diet and pair-fed groups, the number of taste buds per papilla (per animal) and the average profile area of the taste bud were significantly smaller than those of the corresponding controls (p < 0.05). Ultrastructural changes were seen only in the taste buds of weanling rats fed the zinc-deficient diet, with derangement of the architecture of the taste bud and widening of the intercellular space between taste bud cells. The proportion of type I taste bud cells in the taste buds of weanling rats fed the zinc-deficient diet decreased from 59% to 39%, and that of type II taste bud cells decreased from 25% to 12%. No obvious changes in the ultrastructure of type III taste bud cells were observed. CONCLUSIONS: The main effects of zinc deficiency in weanling and young adult rats and in adequate diet pair-fed rats were changes in the number and size of taste buds, and fine structure changes in the taste bud cells, especially during the accelerated growth stage after weaning.

TUNEL staining and electron microscopy studies of apoptotic changes in the guinea pig vallate taste cells after unilateral glossopharyngeal denervation.

On the basis of our previous report that unilateral glossopharyngeal neurectomy in the guinea pig resulted in degeneration and disappearance of taste buds in ipsilateral vallate papillae (Huang and Lu 1996), it is reasonable to speculate that gustatory denervation may enhance apoptosis of taste bud cells, with taste buds decreasing in number and ultimately disappearing after neurectomy. We were therefore determined to investigate apoptosis of taste bud cells in guinea pig vallate papillae after unilateral glossopharyngeal neurectomy using both terminal deoxynuleotidyl transferase (TdT)-mediated dUTP nick-end labelling (TUNEL) at the light microscopic level and by conventional electron microscopy. A total of 34 adult guinea pigs were unilaterally glossopharyngeal-neurectomized and sacrificed at 3, 6, 12 h and 1, 3 and 7 days after surgery. The results revealed that only a very few TUNEL-positive nuclei indicating apoptosis were present in normal taste buds, but in surgically denervated papillae, they increased in number from 6 h-12 h after surgery, reached at peak on day 1 and then gradually decreased. In apoptotic cells from normal taste buds, electron microscopy revealed condensation of the chromatin against the nuclear envelope, changes in the nuclear envelope, and fragmentation of the nucleus, but the integrity of the plasma membrane and organelles was maintained. Neurectomized taste cells were also characterized by condensed and fragmentary nuclei, compactness of the cytoplasmic organelles, and the appearance of pedunculated protuberances on the cell surface. From these observations, we conclude that: (1) glossopharyngeal neurectomy enhanced apoptosis of vallate taste bud cells in guinea pig; (2) appropriate gustatory nerve innervation is an essential component for the maintenance of the taste bud, and may play a role in apoptosis of taste cells.

Distribution of neuronal intermediate filament proteins in the developing mouse olfactory system.

The distribution of neuronal intermediate filament proteins in the developing mouse olfactory bulb and olfactory epithelium was characterized by immunocytochemical approach. Antibodies against alpha-internexin, neurofilament triplet proteins (NFTPs; NF-L, NF-M, and NF-H) and peripherin were used to determine their expression at different developmental stages. Alpha-internexin and peripherin were first found to be co-localized in the olfactory neuroepithelium during early development. At the perinatal stage, expression patterns of alpha-internexin and peripherin are distinguishable by spatial and temporal manner: peripherin is predominantly expressed in the olfactory nerves; whereas alpha-internexin is expressed in both olfactory nerves and olfactory bulb. Our observation suggests that peripherin as well as alpha-internexin may play some roles in the process formation of olfactory nerves during development. In the developing olfactory periglomerulus, alpha-internexin was found around postnatal Day 3, whereas NFTPs were not observed until postnatal Day 7. Our data showed that the expression of alpha-internexin preceded those of the NFTPs in most neurons of the developing olfactory bulb. Some small neurons in the adult olfactory bulb were uniquely labeled with antibody to alpha-internexin. Our results suggest that alpha-internexin may play a functional role in the neuronal cytoarchitecture of developing olfactory system, and can be a neuronal marker for detecting postmitotic migrating neurons in the adult olfactory bulb.

Morphological changes induced by prostaglandin E in cultured rat osteoblasts.

Prostaglandin E (PGE)-induced morphological changes of osteoblasts and its possible mechanisms were investigated in cultured calvaria and isolated osteoblasts from long bone fragments of neonatal rats. The control osteoblasts, either on the calvaria or isolated from the long bone fragments, were flat, polygonal in shape, and arranged in a monolayer under scanning electron microscopy (SEM) or phase contrast microscopy. Treatment with 1 mumol/L of prostaglandin E2 (PGE2, 2 h) caused these bone cells to contract a soma, whereas 10 and 100 mumol/L PGE2 (2 h) caused 18%-30% of the bone cells to elongate and expose the undersurface. Incubation of the cultured osteoblasts with PGE2 at different time periods showed a bell-shaped pattern with the optimal response at 2 h of incubation. A similar reaction can be induced by treatment with prostaglandin E1 (PGE1) or dibutyryl cyclic adenosine monophosphate (DBcAMP) in combination with 3-isobutyl-1-methylxanthine (IBMX). Furthermore, we assessed the percentage of responsive isolated bone cells to investigate interactions with other agents. The morphological changes induced by PGEs were inhibited by H-8, a protein kinase inhibitor. On the other hand, elevated intracellular calcium enhanced the PGE-induced morphological changes. Fluorescence labeling showed that PGEs caused the breakdown of the actin microfilaments, but spared the microtubules and vimentin filaments in the isolated osteoblast-like cells. These results suggest that the morphological changes of osteoblasts induced by PGEs may be related to the intracellular cAMP and calcium levels.

Biochemical and histopathological changes in the mortality caused by acute ischemic limb injury: a rabbits' model.

Restoration of blood flow to an acute ischemic extremity may deteriorate the ischemic injury, lead to multiple organ dysfunction or even death. This paradox of continuing injury during reperfusion is not completely understood. The role of multi-organ damage in the mortality caused by ischemic limb injury is also still not clarified. The purpose of this study is to determine the biochemical and histopathological changes in the mortality caused by ischemic limb injury. After anesthesia, the hindlimbs of 14 New Zealand white rabbits were made ischemic and set into 8 hours or 12 hours of ischemia. Blood samples were obtained then the creatine kinase (CK) levels were determined and CK isoenzymes analyzed. All rabbits with 8 hours' ischemia survived well, and 5 of the 7 rabbits with 12 hours' ischemia expired within 8 hours after reperfusion. CK elevation was correlated most strongly with the time of the ischemic insults. The percentage of CK-MB isoenzyme remained unchanged after 8 hours' ischemia-reperfusion insult, while increased significantly after 12 hours' ischemia-reperfusion insult. Histologic examinations showed that the major systemic manifestation was massive destruction of the liver and kidney. The injuries are more obvious in areas with the greatest blood flow during reperfusion. We concluded that the ratio of CK-MB isoenzyme is most useful for distinguishing the risk of mortality caused by acute ischemic limb injury, and the cause of systemic complications are attributed to the multi-organ failure.

Inhibition of the Raf-1 kinase by cyclic AMP agonists causes apoptosis of v-abl-transformed cells.

Here we investigate the role of the Raf-1 kinase in transformation by the v-abl oncogene. Raf-1 can activate a transforming signalling cascade comprising the consecutive activation of Mek and extracellular-signal-regulated kinases (Erks). In v-abl-transformed cells the endogenous Raf-1 protein was phosphorylated on tyrosine and displayed high constitutive kinase activity. The activities of the Erks were constitutively elevated in both v-raf- and v-abl-transformed cells. In both cell types the activities of Raf-1 and v-raf were almost completely suppressed after activation of the cyclic AMP-dependent kinase (protein kinase A [PKA]), whereas the v-abl kinase was not affected. Raf inhibition substantially diminished the activities of Erks in v-raf-transformed cells but not in v-abl-transformed cells, indicating that v-abl can activate Erks by a Raf-1-independent pathway. PKA activation induced apoptosis in v-abl-transformed cells while reverting v-raf transformation without severe cytopathic effects. Overexpression of Raf-1 in v-abl-transformed cells partially protected the cells from apoptosis induced by PKA activation. In contrast to PKA activators, a Mek inhibitor did not induce apoptosis. The diverse biological responses correlated with the status of c-myc gene expression. v-abl-transformed cells featured high constitutive levels of expression of c-myc, which were not reduced following PKA activation. Myc activation has been previously shown to be essential for transformation by oncogenic Abl proteins. Using estrogen-regulated c-myc and temperature-sensitive Raf-1 mutants, we found that Raf-1 activation could protect cells from c-myc-induced apoptosis. In conclusion, these results suggest (i) that Raf-1 participates in v-abl transformation via an Erk-independent pathway by providing a survival signal which complements c-myc in transformation, and (ii) that cAMP agonists might become useful for the treatment of malignancies where abl oncogenes are involved, such as chronic myeloid leukemias.

Immunohistochemical studies on protein gene product 9.5, serotonin and neuropeptides in vallate taste buds and related nerves of the guinea pig.

The occurrence, distribution and innervation of guinea pig vallate papillae were investigated by means of indirect immunofluorescence and immunoperoxidase methods using antibodies against: a neuron-specific protein, protein gene product 9.5 (PGP 9.5); various neuropeptides including calcitonin gene-related peptide (CGRP), substance P (SP), vasoactive intestinal polypeptide (VIP) and galanin; a monoamine, serotonin (5-hydroxytryptamine; 5HT). Numerous PGP 9.5-immunoreactive nerve fibers were found to form plexuses in the lingual epithelium both intragemmally and extragemmally and to comprise dense bundles in the lamina propria just beneath the epithelium. Moderate numbers of PGP 9.5-immunoreactive cells were observed in the taste buds. These cells, typically spindle in shape, extended through the entire thickness of the taste bud. CGRP-immunoreactive nerve fibers were numerous in the subgemmal connective tissue and entered the epithelium to form intragemmal and extragemmal networks. A dense subgemmal SP-immunoreactive network in the vallate papilla can be linked to the presence of taste buds, even though SP-immunoreactive nerve fibers rarely occurred intragemmally. No taste cells immunoreactive for CGRP and for SP were observed. Immunoreactivity for VIP or galanin was not detected in nerve fibers and taste cells. In contrast, some taste cells and a few, fine networks of nerve fibers in the connective tissue were immunoreactive for 5HT; none of the intraepithelial fibers were 5HT-immunoreactive. We suggest that: 1) functionally, 5HT-containing cells and the CGRP-containing nerve fibers may be primarily involved in the neural transmission or its modulation of the taste sensation; and 2) VIP and galanin can be excluded from that group of substances which plays important roles in taste sensation.

Unilateral innervation of guinea pig vallate taste buds as determined by glossopharyngeal neurectomy and HRP neural tracing.

The innervation pattern by primary afferent nerve fibres and the neurotrophic effect on taste cells were investigated in the guinea pig vallate taste bud by means of glossopharyngeal neurectomy and horseradish peroxidase (HRP) or wheat germ agglutinin-horseradish peroxidase (WGA-HRP) tracing. In the glossopharyngeal neurectomy study, taste buds in the vallate papillae of adult guinea pigs were denervated by unilateral resection of the right glossopharyngeal nerve. Denervated animals were killed on days 1, 3 and 5 and weeks 1-9, 12 and 24 postneurectomy. The results showed that, on the denervated side, the taste buds decreased significantly in number during the 1st 2 wk, and disappeared completely by wk 3; no mature taste buds were present even 24 wk after neurectomy. This suggests that the vallate taste buds disappear in the absence of the glossopharyngeal nerve. In the neural tracing study, HRP or WGA-HRP was injected into the proximal end of the right glossopharyngeal nerve, near the jugular foramen. After a survival time of 24 h, the vallate papillae were sectioned and examined by light and electron microscopy. Light microscopy revealed that the HRP or WGA-HRP-labelled fibres innervated the vallate taste buds of the injected side. Most of the taste cells in the buds were labelled with HRP or WGA-HRP reaction products from the basal to the apical region. At the ultrastructural level, the reaction products were confined to the cytoplasm of the labelled cells, which were identified as type I, II and III cells, but not basal cells. Labelled intragemmal nerve profiles were seen among the taste cells. No synapse formation was seen with nerve profiles abutting on type I and II cells, whereas on certain type III cells, typical synapses were formed. We conclude that both the right and left vallate papilla in the guinea pig are unilaterally innervated by the glossopharyngeal nerve without cross-innervation.

Differential permeability of blood microvasculatures in various sympathetic ganglia of rodents.

The ultrastructure of capillaries and their permeability to lanthanum ion and horseradish peroxidase (HRP) in various rodent sympathetic ganglia were investigated in this study. Electron microscopic observation revealed that most capillaries surrounding the principal neurons in these ganglia were of the continuous (non-fenestrated) type, while the fenestrated capillaries were consistently associated with the small granule-containing (SGC) cells in rat and hamster superior cervical ganglia and the coeliac-mesenteric ganglia (CMG) complex. Both of the capillaries surrounding the principal neurons and adjacent to SGC cells in various gerbil sympathetic ganglia or in rat and hamster thoracic ganglia were of the non-fenestrated type. After lanthanum perfusion, lanthanum tracer was limited to the blood-vessel lumen but was apparently obstructed by the tight junctions of capillaries. No lanthanum was visible in the extravascular space surrounding the principal neurons of rodent superior cervical and thoracic ganglia. By contrast, lanthanum extravasation was observed in the luminal, abluminal and perivascular surface of capillaries in the CMG complex and near SGC cells in the superior cervical ganglion. Injecting HRP showed that all blood vessels in various sympathetic ganglia were impermeable to HRP. HRP-DAB reaction product was limited to the lumen of capillaries, blocked by tight junctions and obstructed by fenestral diaphragms of fenestrated capillaries close to SGC cells. We conclude that: (1) the capillaries surrounding the principal neurons in rodent superior cervical and thoracic ganglia are more restrictive to HRP and lanthanum ion than those anywhere in the CMG complex or in regions containing SGC cells of superior cervical ganglia; (2) according to the results of lanthanum and HRP experiments, the existence of different blood-barrier properties are present among different rodent sympathetic ganglia or within the same ganglion.

Ultrastructural studies on myofibrillogenesis and neogenesis of skeletal muscles after prolonged traction in rabbits.

Little is known about the morphological response of muscle after long term traction. The purpose of this study was to investigate the morphological changes of skeletal muscle during limb lengthening. After application of mini-extraskeletal fixator, the hindlimb of New Zealand white rabbit was osteotomized and then slowly lengthened at the rate of 1 mm/day up to a 20 mm gain in length. The muscles of hindlimbs were perfused and dissected. Morphological studies were performed at electron microscopic level. Transmission electron microscopy revealed foci of microtrauma at the myotendinous junction. The distance between the muscle fibers and tendon parenchyma increased, with numerous primitive mesenchyme-like cells interposed within this gap. The cytoplasmic space of these cells was devoid of myofibril formation at the ends of stretched fibers. Within the satellite near the myotendinous junction myofilament production was observed in various gradations of maturation. It is concluded that myofibrillogenesis with traction neogenesis of skeletal muscle during limb lengthening does exist and occurs mainly near the myotendinous junction. The myotendinous junction in mature skeletal muscle actively participated in the process of limb lengthening.

Biomechanical properties of the triceps surae muscle after limb lengthening.

The hindlimb of New Zealand white rabbit was osteotomized and then slowly lengthened at a rate of 1 mm/day until a 2.0-cm gain in length was reached. The triceps surae muscle-tendon unit was then tested either with or without 2 months fixation for bony consolidation. After limb lengthening, the strain at peak load was decreased and the axial rigidity was increased significantly; but other biomechanical parameters such as peak load, maximal deformation at peak load, stiffness, and energy absorption before peak load did not show any significant difference. We conclude that healthy triceps surae muscle has a great potential to be lengthened despite the changes in intrinsic structural properties of the muscle. The change in the biomechanical properties occurred during the time of distraction and was not affected by the time of bony consolidation. RELEVANCE: As noted in this study, many biomechanical parameters of the triceps surae muscle were not affected after prolonged distraction. This fact suggested that the healthy muscle-tendon unit is not a main causative factor of joint contracture after limb lengthening. However, there were intrinsic changes in the structural properties of the triceps surae muscle. Equinus contracture after limb lengthening may be caused by the aggravated intrinsic structural changes of the injured or denervated scarred muscular tissue.

Investigation of the blood-ganglion barrier properties in rat sympathetic ganglia by using lanthanum ion and horseradish peroxidase as tracers.

Vascular permeability in various rat sympathetic ganglia, including superior cervical ganglia, thoracic ganglia and the celiac-mesenteric ganglia (CMG) complex, was investigated by using lanthanum and horseradish peroxidase (HRP) as tracers with special attention to the neuronal and small granule-containing (SGC) cell area. After lanthanum perfusion, lanthanum tracer was present within the lumen of blood vessels. No lanthanum depositions were found in the extravascular space surrounding neurons in the superior cervical and thoracic ganglia. By contrast, an accumulation of lanthanum was observed in both luminal, abluminal and subendothelial surface of blood vessels in neuronal and SGC cell areas of the CMG complex and surrounding SGC cells in superior cervical ganglia. Injecting HRP revealed that all blood vessels of various sympathetic ganglia, either in neuronal or in SGC cell areas, were impermeable to HRP. HRP reaction product was limited to the vascular lumen and macrophages. The escape of HRP was obstructed by the junctional complex at intercellular clefts of endothelia and also by the diaphragms of the fenestrated capillaries associated with SGC cells. We conclude that there are different properties in the blood-ganglion barriers among rat sympathetic ganglia: (1) continuous capillaries in superior cervical ganglia and thoracic ganglia provide an efficient blood-ganglion barrier that prevents the penetration of tracers, and (2) capillaries in the CMG complex and in regions of the superior cervical ganglia that contain SGC cells possess a selective blood-ganglion barrier that discriminates between tracers based on their molecular sizes.

Ultrastructure, ZIO-staining and chromaffinity of gerbil pinealocytes.

The ultrastructure and cytochemistry of the gerbil pineal gland were studied by the conventional electron microscopy, zinc iodide-osmium tetroxide (ZIO) staining and chromaffin reaction. Conventional electron microscopy revealed that the ultrastructure of gerbil pinealocytes are similar to other rodents, i.e., irregular cell contour with numerous cytoplasmic processes, round or oval nucleus and prominent nucleoli, elongated mitochondria with flattened and tubular cristae and dense matrix, well-developed Golgi apparatus and its associated structures, abundant elements of endoplasmic reticulum--both smooth and rough varieties, and bundles of microfilament and microtubule in the cytoplasm. Some pinealocyte processes contain numerous small clear and "slightly coated" vesicles. Numerous profiles of varicosities containing small dense-cored and clear vesicles were frequently encountered. After ZIO treatment, ZIO staining was preferentially localized in the cytoplasm of some, but not all, of the gerbil pinealocytes. Numerous small clear vesicles (30-50 nm in diameter) in the process of the pinealocytes or in the varicosities of the nerve fibers showed strong ZIO-philia. After chromaffin reaction treatment, the number and electron density of small clear and dense-cored vesicles in the profiles of nerve varicosities increased and this indicates that some of the small clear and dense-cored vesicles in the varicosities are reactive. It is thus concluded that (1) the vesicles in the pinealocytes may be rich in cystine and/or cysteine and possibly the organelle is involved in the sequestering calcium ion during the calcification of the pineal concretions, and (2) the small dense-cored and clear vesicles in the nerve fibers in the gerbil pineal parenchyma may contain both serotonin and primary biogenic amines.

ZIO impregnation and cytochemical localization of thiamine pyrophosphatase and acid phosphatase activities in small granule-containing (SGC) cells of rat superior cervical ganglia.

Cytochemical relationship between Golgi complex and dense-cored granules (DCGs) of small granule-containing (SGC) cells in rat superior cervical ganglia was examined in electron microscopy by zinc-iodide-osmium tetroxide (ZIO) method and by enzyme cytochemistry for thiamine pyrophosphatase (TPPase) and acid phosphatase (ACPase). After ZIO impregnation, all the saccules of Golgi apparatus and some of tubular rough endoplasmic reticulum (rER) were stained. DCGs in periphery of SGC cells were not stained, but varying degrees of dense deposits occurred in the DCGs in vicinity of Golgi trans-saccules. Both TPPase and ACPase activities were localized in one or two stacked layers of saccules on the trans side of the Golgi complex. No reaction products were demonstrated in the DCGs. From these results, we suggest that the DCGs of SGC cells in rat superior cervical ganglia are derived from the Golgi complex, and that lysosomal cleavage of protein contents in the DCGs may occur in the trans Golgi saccules.

Analysis of neogenesis in rabbit skeletal muscles after chronic traction.

After application of a modified Orthofix mini-extraskeletal fixator, the hind limb of New Zealand White rabbits was osteotomized and then slowly lengthened at a rate of 1 mm/day. After a 20 mm gain in length, the net weight and the length of muscular and tendinous portions were measured and histological examination was carried out in triceps surae muscles. Quantitative analysis showed a significant increase in the gained length of the muscular portion (28.05% to 30.65%). Histological studies of these lengthened muscles showed a generalized increase in cellularity with scanty inflammatory cell infiltration near the myotendinous junction. The increased cellularity is due to the presence of muscle precursor cells characterized by large, oval and pale-stained vesicular nuclei and two prominent nucleoli. The nuclei of these precursor cells were larger and more numerous near the myotendinous junction, and gradually changed into a flattened and more condensed form at a distance from the junction. Occasionally, chains of centrally-located nuclei of primitive myoblasts were also visible. It is concluded that traction neogenesis of the skeletal muscle during limb lengthening does exist and occurs mainly near the myotendinous junction.