Axotomy along with hypoxia enhances the neuronal NADPH-d/NOS expression in lower brain stem motor neurons of adult rats.

This study was aimed to determine whether axotomy coupled with hypoxia would exert a more profound effect on injury-induced neuronal nitric oxide synthase (NOS) expression. In this connection, the vagus and the hypoglossal nerves of adult rats were transected unilaterally in the same animal, and half of the operated animals were subjected to hypoxia treatment. Both the neuronal NOS immunohistochemistry and the nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry were used to assess the neuronal NOS expression. The present results have shown that the number of NADPH-d/NOS-positive [NADPH-d/NOS(1)] neurons in the hypoglossal nucleus (HN) peaked at 14 days after axotomy, while that in dorsal motor nucleus of vagus (DMN) and nucleus ambiguus (NA) was progressively increased up to 60 days. The up-regulation of NADPH-d/NOS in HN and DMN was more pronounced in hypoxic than in normoxic animals, a feature that was not evident in the NA. Quantitative analysis showed that the number of surviving motoneurons in normoxic animals was significantly higher than those subjected to hypoxia at 14 days postaxotomy in HN and at all postaxotomy time points in DMN. The difference may be attributed to their different functional components. Since O2 deprivation leads to poor cellular function, the stronger expression of NADPH-d/NOS and the more drastic neuronal loss following nerve transection in the hypoxic animals compared with the controls suggest that hypoxia plays an important role in peripheral neuropathies in which NO is implicated.

The distribution and characterization of NADPH-d/NOS-IR neurons in the rat cuneate nucleus.

Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and nitric oxide synthase (NOS) immunohistochemistry have been used to characterize the nitric oxide (NO)-containing neurons in the rat cuneate nucleus. The present results showed that NADPH-d-positive/NOS-immunoreactive (-IR) neurons were distributed in the entire rostrocaudal extent of the nucleus. In the caudal region (approximately 1-2 mm caudal to the obex), NADPH-d/NOS-IR neurons were aggregated along the dorsal area of the nucleus notably in the lateral aspect. When traced rostrally, labeled neurons were progressively reduced and the cells were randomly distributed. The labeled neurons varied from round, ovoid to spindle-shaped with a mean profile area of about 140.1+/-1.7 microm(2) (n=720). They made up 7-10% of the neuronal population in the cuneate nucleus. By immunoelectron microscopy, the immunoreaction product was deposited throughout the cytoplasm extending from the soma to the proximal and distal dendrites. Results of NADPH-d staining paralleled that of NOS immunohistochemistry. Furthermore, NADPH-d reactivity and NOS-IR were colocalized in the same neurons following double labeling. Using NADPH-d histochemistry along with anti-gamma-aminobutyric acid (GABA) and -glycine postembedding immunolabeling for identification of GABA- and glycine-IR neurons, respectively, about 33% of the NADPH-d-positive neurons contained both GABA and glycine, 26% of them contained only glycine, while 41% of them showed neither GABA nor glycine labeling. Cuneothalamic neurons (CTNs) were identified by injecting the retrograde tracer Fluorogold (FG) into the ventrobasal complex of the thalamus. Numerous FG-labeled neurons were present in the contralateral cuneate nucleus, but none were reactive for NADPH-d. The present results suggest that approximately 60% of the NADPH-d/NOS-IR neurons in the cuneate nucleus are interneurons containing GABA and/or glycine.

Somatic noxious mechanical stimulation induces Fos expression in the postsynaptic dorsal column neurons in laminae III and IV of the rat spinal dorsal horn.

This study was conducted to ascertain the possible expression of Fos-like immunoreactivity (Fos-LI) in the postsynaptic dorsal column (PSDC) neurons in response to noxious mechanical stimulation of the forepaw glabrous area of normal rats. For this purpose, Fos immunohistochemistry along with Fluoro-Gold (FG) retrograde tracing was utilized. After repeated noxious pinching of the forepaw glabrous area, there was a marked increase in number of Fos-LI neurons in the dorsal horn, including Rexed's laminae III and IV, at C5-T1 spinal cord segments ipsilateral to the stimulation. Between segments C5 and T1, about 40% of the Fos-LI neurons in laminae III and IV were distributed at segment C7. In the rats subjected to the noxious pinch coupled with FG injection into the right cuneate nucleus, PSDC neurons double labeled with Fos and FG were localized in the ipsilateral laminae III and IV extending from segment C5 to T1, with about 70% of them distributed at segments C6 and C7. At segment C6 or C7, double-labeled neurons made up about 10% of the PSDC neurons that projected their axons to the cuneate nucleus. Most of the double-labeled neurons appeared fusiform with their primary dendrites projected dorso-ventrally. The present results suggest that the morphologically distinct, subclasses of PSDC neurons in spinal laminae III and IV may contribute to the central transmission of mechanical nociceptive information through the dorsal column into the cuneate nucleus.

Cadaveric study of blood supply to the lower intraorbital fat: etiologic relevance to the complication of anaerobic cellulitis in orbital floor fracture.

BACKGROUND AND PURPOSE: Although orbital fractures are common, orbital cellulitis rarely develops following orbital fracture. We hypothesized that compromise of the blood supply to the intraorbital fat during orbital floor fracture is responsible for this condition. The purpose of this study was to determine whether or not the lower intraorbital fat is supplied by a branch of the infraorbital artery along the orbital groove or canal on the orbital floor. MATERIALS AND METHODS: We dissected 14 orbits from seven fixed human cadavers and 12 orbits from six fresh cadaver heads following dye injection into the maxillary artery. The sites of dye-filled vessels branching from the infraorbital artery supplying the lower intraorbital fat were measured and plotted on a two-dimensional orbital floor graph. RESULTS: A main branch of the infraorbital artery rose through the medial orbital floor to supply the lower intraorbital fat in all of the cadaver orbits. The sites of the branching point of the vessel ranged from 0 to 5 mm (mean, 2.2 mm; n = 14) medial to the line connecting the infraorbital foramen and the infraorbital groove. The shortest distance measured from the branching point to the orbital rim ranged from 3 to 20 mm (mean, 14.1 mm; n = 14). This suggests that if orbital fracture were to occur around the infraorbital groove or canal, this vascular pedicle would be in danger of being incarcerated by bone fragments. CONCLUSION: Our cadaveric investigation revealed that the lower intraorbital fat is supplied by a branch of the infraorbital artery along the infraorbital groove or canal on the orbital floor. This finding suggests that compromised blood supply to the intraorbital fat may cause anaerobic cellulitis or enophthalmos.

Afferent synaptic contacts on glycine-immunoreactive neurons in the rat cuneate nucleus.

This study was aimed to clarify whether the primary afferent terminals (PATs), GABAergic terminals, and glutamatergic terminals made direct synaptic contacts with glycine-IR neurons in the cuneate nucleus of rats. In this connection, injection of the anterograde tracer WGA-HRP into brachial plexus, antiglycine preembedding immunoperoxidase, and anti-GABA, along with antiglutamate postembedding immunogold labeling, were used to identify the PATs, glycine-IR neurons, GABA-IR terminals, and glutamate-IR terminals, respectively. The present results showed that HRP-labeled PATs, immunoperoxidase-labeled glycine-IR terminals, immunogold-labeled GABA-IR, and glutamate-IR terminals made axodendritic synaptic contacts with immunoperoxidase-labeled glycine-IR neurons. The latter three presynaptic elements also formed axosomatic synapses with glycine-IR neurons. Statistical analysis has shown that the minimum diameter of the glycine-IR dendrites postsynaptic to the above-mentioned four presynaptic elements did not differ significantly. In addition, the synaptic ratio of the glutamate-IR terminals on the glycine-IR dendrites was higher than that of GABA-IR terminals. The synaptic ratio of the GABA-IR terminals on glycine-IR dendrite was in turn higher than that of the PATs and glycine-IR terminals. It is suggested that the PATs and glutamate-IR terminals on the glycine-IR neurons may be involved in subsequent postsynaptic inhibition for spatial precision of lateral inhibition. On the other hand, the GABA-IR and glycine-IR terminals which make synaptic contacts with the dendrites of glycine-IR neurons may provide a putative means for disinhibition or facilitation to maintain the baseline neuronal activity in the rat cuneate nucleus. The results of quantitative analysis suggest that glutamate act as the primary excitatory neurotransmitter, while GABA, when compared with glycine, may serve as a more powerful inhibitory neurotransmitter on glycine-IR neurons in the rat cuneate nucleus.

Evidence of neuroanatomical connection between the superior cervical ganglion and hypoglossal nerve in the hamster as revealed by tract-tracing and degeneration methods.

Previous studies have shown the existence of a sympathetic component in some cranial nerves including the hypoglossal nerve. In this study, the horseradish peroxidase (HRP) tract-tracing retrograde technique and experimental degeneration method were used to elucidate the possible neuroanatomical relationship between the superior cervical ganglion (SCG) and the hypoglossal nerve of hamsters. About 10% of the SCG principal neurons were HRP positive following the tracer application to the trunk of hypoglossal nerve. Most of the HRP-labelled neurons were multipolar and were randomly distributed in the ganglion. When HRP was injected into the medial branch of the hypoglossal nerve, some of the SCG neurons were labelled, but they were not detected when HRP was injected into the lateral branch. The present findings suggest that postganglionic sympathetic fibres from the SCG may travel along the hypoglossal nerve trunk via its medial branch to terminate in visceral targets such as the intralingual glands. By electron microscopy, the HRP reaction product was localised in the neuronal somata and numerous unmyelinated fibres in the SCG. In addition, HRP-labelled axon profiles considered to be the collateral branches of the principal neurons contained numerous clear round and a few dense core vesicles. Besides the above, some HRP-labelled small myelinated fibres, considered to be visceral afferents, were also present. Results of experimental degeneration following the severance of the hypoglossal nerve showed the presence of degenerating neuronal elements both in the hypoglossal nucleus and the SCG. This confirms that the hypoglossal nerve contains sympathetic component from the SCG which may be involved in regulation of the autonomic function of the tongue.

A beta-fiber intensity stimulation of chronically constricted median nerve induces c-fos expression in thalamic projection neurons of the cuneate nucleus in rats with behavioral signs of neuropathic pain.

This study was aimed to investigate the possible involvement of neurons in the cuneate nucleus (CN) in the processing of A beta afferent inputs evoked by electrical stimulation of constricted median nerve in rats with behavioral signs of neuropathic pain. Immunohistochemical localization of Fos protein was used to examine the neuronal activation, and the combination of Fos immunohistochemistry with the retrograde labeling of Fluoro-Gold (FG) injected into the ventrobasal complex of the thalamus was used to characterize the activated neurons. Two weeks after unilateral median nerve constriction injury, the rats exhibited behavioral signs of neuropathic pain in the affected forepaws. In rats after nerve injury but without electrical stimulation, some Fos-like immunoreactive (Fos-LI) neurons were detected in the dorsal horn of the seventh cervical segment (C7) but none was found in the CN. Similar features were also noted when the stimulation of the intact median nerve served as an additional control. After A beta-fiber intensity stimulation of the previously constricted median nerve, an increase in number of Fos-LI neurons occurred in the medial half of the ipsilateral C7 dorsal horn as well as in the ipsilateral CN. In the latter, the Fos-LI neurons were located in the median nerve projection territory throughout the nucleus. Most of the Fos-LI neurons were distributed in the middle region of the CN, with about 78% of them emitting FG fluorescence indicating that they were cuneothalamic projection neurons. The results of this study suggest that the dorsal column-medial lemniscal system may contribute to the transmission and modulation of A beta-fiber mediated neuropathic pain signals.

Ultrastructural localisation of NADPH-d/nNOS expression in the superior cervical ganglion of the hamster.

This study examined NADPH-d and nNOS expression in the SCG of hamsters. By light microscopy, numerous NADPH-d/NOS positive processes were widely distributed in the ganglion. Ultrastructurally, the NADPH-d reaction product was associated with the membranous organelles of neuronal soma, dendrites, myelinated fibres, small granular cells, and axon profiles bearing agranular vesicles. The NOS immunoreaction product, on the other hand, was localised in the cytoplasm of principal neurons and dendrites. Some of the NADPH-d/NOS labelled processes formed junctional contacts including synapses or zonulae adherentia. Compared with the neurons, the nonneuronal cells in the ganglion, namely, macrophages, satellite cells and endothelial cells were labelled by NADPH-d but devoid of nNOS immunoreaction product. The results suggest that the NADPH-d/NOS positive fibres in the SCG originate not only from the projecting fibres of the lateral horns of thoracic spinal cord, but also from the principal neurons and small granular cells; some may represent visceral afferent fibres. Electron microscopic morphometry has shown that about 67% of the principal neurons contain NADPH-d reaction product, and that the majority were small to medium sized neurons based on cross-sectional areas in image analysis. On the basis of the present morphological study, it is concluded NO is produced by some local neurons and possibly some nonneuronal cells in the SCG as well as some fibres of extrinsic origin. In this connection, NO may serve either as a neurotransmitter or neuromodulator.

Somatostatin-IR neurons are a major subpopulation of the cuneothalamic neurons in the rat cuneate nucleus.

This study was aimed to localize and characterize the somatostatin-immunoreactive (SOM-IR) neurons in the rat cuneate nucleus (CN). By immuno-histochemistry, the SOM-IR neurons, which were widely distributed in the nucleus, were round, spindle or multiangular in shape (mean area = 226.1 +/ -3.1 microm(2), n = 1016). By electron microscopy, the neurons shared all the ultrastructural features of the cuneothalamic neurons (CTNs) which showed a slightly indented nucleus and a fairly rich cytoplasm containing well-developed Golgi apparatuses and rough endoplasmic reticulum (rER). The SOM immunoreaction product filled the cytoplasm of the neurons extending from the soma to the proximal and distal dendrites, which were postsynaptic to unlabeled boutons. In addition to soma and dendrites, SOM-IR boutons were also identified which made axodendritic synaptic contacts with SOM-IR dendrites. The SOM-IR neurons were characterized by using anti-SOM pre-embedding immunolabeling coupled with horseradish peroxidase (HRP) retrograde method, or SOM immunolabeling along with anti-glutamate, gamma-aminobutyric acid (GABA) or glycine post-embedding immunolabeling for identification of CTNs, glutamate-IR, GABA-IR and glycine-IR neurons, respectively. It was shown that more then 80% of the CTNs contained SOM and, furthermore, they contained glutamate but not GABA or glycine. On the basis of present findings, it is suggested the majority of the SOM-IR neurons in the rat CN are CTNs and that they may be involved in modulation of somatosensory synaptic transmission.

Melatonin attenuates neuronal NADPH-d/NOS expression in the hypoglossal nucleus of adult rats following peripheral nerve injury.

Oxidative stress and massive production of nitric oxide (NO) have been implicated in the neuropathogenesis following peripheral nerve injury. This study was aimed to ascertain whether melatonin would exert its neuroprotective effect on the lesioned hypoglossal neurons after peripheral axotomy, since it is known to reduce the oxidative damage in a variety of experimental neuropathologies in which NO is involved. Right-sided hypoglossal nerve transection was performed in adult rats following which the animals were given two different doses of melatonin administered intraperitoneally for 3, 7, 14, 21 and 30 successive days. Nicotinamide adenine dinucleotide phosphate-diaphorase (NADPH-d) histochemistry and neuronal nitric oxide synthase (nNOS) immunohistochemistry were carried out to detect the neuronal NADPH-d/NOS expression in the hypoglossal nucleus (HN). At various time intervals following axotomy, the neurons in the affected HN were induced to express NADPH-d/NOS reactivity on the lesioned side peaking at 14 days. However, the enzyme expression was markedly depressed by melatonin treatment in a dose-dependent manner in terms of frequency of labelled neurons and staining intensity. It is suggested that the suppressive effect of melatonin on NADPH-d/NOS expression may be attributed to its antioxidant properties. Hence, in consideration of therapeutic strategies for reducing the oxidative stress following peripheral nerve injury, melatonin may prove to be beneficial.

Cuneothalamic relay neurons are postsynaptic to glycine-immunoreactive terminals in the rat cuneate nucleus.

This study was aimed to clarify whether the cuneothalamic relay neurons (CTNs) in the rat cuneate nucleus contained glycine or whether the neurons were modulated directly by presynaptic glycine-IR terminals. For this purpose, retrograde transport of wheat germ agglutinin conjugated with horseradish peroxidase (WGA-HRP) and immunoperoxidase labelling for glycine have been used to ascertain if the CTNs in the rat are glycine-immunoreactive (glycine-IR). Our results have shown that the WGA-HRP-labelled CTNs (mean area = 318 +/- 6.5 microm(2)) were not reactive for glycine. Glycine immunoreactivity, however, was localized in some small-sized neurons (mean area = 210 +/- 6.2 microm(2)) and axon terminals associated with the CTNs. The synaptic organization between the glycine-IR terminals and CTNs was further analyzed using anti-glycine postembedding immunogold labelling. By electron microscopy, the immunogold-labelled glycine-IR terminals containing pleomorphic synaptic vesicles formed symmetrical synaptic contacts with the dendrites, dendritic spines, and somata of CTNs. Quantitative estimation showed that the mean ratios of glycine-IR terminals to total terminals associated with the soma, proximal dendrites and distal dendrites of the CTN were 49.5, 45.2, and 45.8%, respectively. The higher incidence of glycine-IR terminals on the soma, however, was not significantly different from that of the proximal and distal dendrites. Notwithstanding the above, this study has shown a large number of glycine-IR terminals making direct synaptic contacts with CTNs, suggesting that glycine is one of the important neurotransmitters involved in postsynaptic inhibition on the cuneothalamic relay neurons to modulate incoming somatosensory information from forelimb areas in the rat.

Ultrasonographic measurement of the mechanical properties of the sole under the metatarsal heads.

The sole under the metatarsal heads functions as a shock absorber during walking and running. The mechanical properties of the sole provide the primary defense against the development of metatarsalgia and foot ulceration. However, limited information about these properties has been documented. In this study, we used ultrasonography to evaluate the mechanical properties, including unloaded thickness, compressibility index, elastic modulus, and energy dissipation ratio, of the sole in 20 healthy subjects. The unloaded thickness decreased progressively from the first to the fifth metatarsal heads, with values of 1.50, 1.36, 1.25, 1.14, and 1.04 cm. The sole under the first metatarsal head had the greatest values for the compressibility index and elastic modulus (55.9% and 1.39 kg/cm2), and the sole under the third metatarsal head had the smallest values (50.8% and 1.23 kg/cm2). The sole under the fifth metatarsal head had the greatest energy dissipation ratio (33.7%), followed by that under the third, second, first, and fourth metatarsal heads. Multivariate adjusted linear regression showed that the unloaded thickness, compressibility index, and elastic modulus values increased significantly with age and body weight (p < 0.05) and that the energy dissipation ratio increased significantly with body weight (p < 0.05)

Sonographic detection of occult fractures in the foot and ankle.

PURPOSE: The purpose of this retrospective study was to determine whether high-resolution sonography can aid in the diagnosis of radiographically occult fractures in the foot and ankle. METHODS: High-resolution sonography with a 10-MHz linear-array transducer was performed in 268 patients with foot and ankle injuries whose initial plain x-ray films were negative for fracture. RESULTS: Twenty-four patients had occult fractures demonstrated by sonography. On sonography, the occult fractures appeared as a discontinuity of cortex echogenicity. The fractures were found at the calcaneus (n = 8), metatarsus (n = 6), talus (n = 3), navicular bone (n = 3), cuboid bone (n = 2), cuneiform bone (n = 1), and lateral malleolus (n = 1). Review of the patients' radiographs revealed tiny fractures at the sonographically identified locations in 2 patients. The first 5 patients underwent bone scans, which confirmed the presence of the fractures. The first 11 patients received follow-up sonographic examination 6 weeks after diagnosis; in all 11, an echogenic line over the previous fracture site, presumably representing callus formation, was noted. CONCLUSIONS: Sonography-a readily available, noninvasive imaging technique-can provide important information about soft tissue injuries and cortical discontinuities in the foot and ankle area. Using this procedure, occult fractures can be identified and delineated, and costly procedures such as MRI can be avoided.

Sonographic evaluation of the posterior cruciate ligament in amputated specimens and normal subjects.

The purpose of this study was to define the sonographic appearance and echogenicity of the normal posterior cruciate ligament. We examined the posterior cruciate ligament of five amputated specimens and five normal subjects using a 10 MHz linear array transducer. One K-wire was inserted into the substance of the posterior cruciate ligament of the amputated knee specimens to verify the location of the ligament on the sonogram. Various angles of insonation were used to examine the echogenicity of the posterior cruciate ligament. The results showed that the in situ posterior cruciate ligament appeared as a hypoechoic band relative to the surrounding tissue on sonograms, but it appeared hyperechoic when it was isolated and immersed in a water bath. The specific spatial orientation of the posterior cruciate ligament and anisotropy phenomenon contributed to the hypoechogenicity of the posterior cruciate ligament in situ on sonogram.

Ultrasonographic assessment of posterior heel pain.

To investigate the value of ultrasonography in the diagnosis of posterior heel pain, 68 patients with normal plain x-ray findings of the posterior heel underwent ultrasonographic examination with a 10-MHz linear array probe. The findings included Achilles tendinosis (31 patients), retrocalcaneal bursitis (12), superficial Achilles bursitis (7), soft tissue mass (7), Achilles tendon rupture (4), xanthoma (3), tenosynovitis of the flexor hallucis longus tendon (2), and negative findings (2). Sixteen of these patients underwent surgery after ultrasonographic examination. The surgical diagnoses were consistent with the ultrasonographic diagnoses in all cases. With high-resolution ultrasonography, pathologic conditions of the posterior heel can be readily differentiated.

Noninvasive assessment of the viscoelasticity of peripheral arteries.

Currently used methods of examining the mechanical properties of blood vessel walls are either indirect or invasive, or measure vessel diameter and pressure waveforms at different sites. We developed a noninvasive technique to assess the mechanical properties and viscoelasticity of peripheral arteries. The pressure-strain elastic modulus (Ep) and the viscoelastic properties (energy dissipation ratio, EDR) of the common carotid artery (CCA), brachial artery (BA), radial artery (RA) and dorsalis pedis artery (DPA) were determined by means of palpating pressure and diameter distension waveforms extracted from high-resolution ultrasonography. The methodology was validated in vitro using an elastic tube phantom, as well as in vivo. In vivo study in 10 healthy volunteers (mean age 22 y) showed that the pressure-diameter curves were nonlinear, with an inflection at about 85-90 mmHg, and routed clockwise with slight hysteresis. The CCA (n = 5) had a mean diameter of 6.74 mm and the pulsatile diameter distension was 12.2%. The Ep calculated at the CCA was 0.44 x 10(6) dyne/cm2 with an EDR of 7.18%. The BA, RA and DPA (n = 10) had mean diameters of 3.91 mm, 2.21 mm and 2.12 mm; arterial strains of 4.60%, 4.25% and 8.91%; mean Ep of 1.39, 1.45, 0.90 x 10(6 )dyne/cm2; and mean EDRs of 6.34%, 6.15% and 5.60%, respectively. The method presented is relatively simple to implement clinically and has potential as a new diagnostic tool for detecting local vascular changes.

Neuronal connections between the auricular skin and the sympathetic pre- and postganglionic neurons of the dog as studied by using pseudorabies virus.

Pseudorabies virus (PrV) as a neuronal tracer was microinjected into the concave surface of the puppy's left pinna to establish the morphological basis of somato-visceral linkage. The virus infected neurons were detected by FITC conjugated with polyclonal swine anti-PrV serum. Labelled neurons were localized in: (1) the trigeminal, geniculate and superior vagal ganglia; (2) the subnucleus caudalis of the spinal trigeminal nucleus; (3) the intermediolateral column (IML) of the thoracolumbar segments and (4) the sympathetic chain ganglia. Present results suggest that when injected into the peripheral nerves, PrV was retrogradely transported to the nerve cell bodies located in the respective sensory ganglia. From the first order sensory neurons, the virus would self-replicate and was transported trans-synaptically via the brainstem nuclei and IML to reach the neurons in the sympathetic ganglia.

Pregnancy achieved by intracytoplasmic sperm injection using cryopreserved vasal-epididymal sperm from a man with spinal cord injury.

Anejaculation and poor semen quality are two major causes of infertility in men with spinal cord injury (SCI). The poor motility of retrieved sperm usually has low fertilization potential and is thought to be unfavorable for cryopreservation. This report describes a pregnancy after intracytoplasmic sperm injection (ICSI) with cryopreserved vasal-epididymal sperm from a man with SCI and anejaculation. An attempt was made to obtain sperm through electroejaculation, but no motile sperm were found in two trials. Therefore, the subject underwent vasal aspiration. The retrieved sperm had a concentration of 26 x 10(6)/mL and a motility of 3%. ICSI was considered to be the best choice for the couple, but the wife did not become pregnant in the first cycle of treatment. A successful pregnancy was achieved by ICSI in the second cycle using frozen-thawed sperm, supernumerary in the previous cycle, with a density of 5 x 10(6)/mL and 1% motility. A set of healthy twins, one boy and one girl, were delivered via cesarean section at 36 weeks of gestation. Complementary to other assisted reproductive techniques, ICSI may provide men with SCI a greater opportunity to father children. The supernumerary sperm, regardless of quality, should be cryopreserved to avoid the necessity and risk of repeated assisted ejaculations and aspirations of the genital tract.

Effects of dexamethasone on antigen expressions and proliferation of amoeboid microglial cells in fetal rat brain.

The present study examined the effect of maternal administration of dexamethasone (DEX) on amoeboid microglial cells (AMC) in fetal rats extending from 16 to 20 days postconception (E16 to E20). After an intraperitoneal injection of DEX into pregnant rats at E10, the external morphology and distribution of immunolabelled AMC as detected with OX-42 and ED1 monoclonal antibodies remained unaltered when compared with those of the controls. The major effect of dexamethasone was on microglial cell population. Thus, with OX-42 and ED1, the numbers of immunolabelled AMC in the intermediate zone lateral to the striatum (IZS) of DEX-treated fetuses which remained relatively unchanged at E16 were significantly reduced at E18. However, OX-42 labelled cells showed an unexpected increase in number at E20 following DEX treatment. Microglial response to DEX was also analyzed in sections stained with the isolectin, GSA I-B4, which specifically binds alpha-D-galactosyl glycoproteins on microglia. The number of GSA I-B4 labelled AMC was significantly increased at E16, declined at E18 and remained constant thereafter in DEX-treated rats when compared with that of the controls. A major finding after DEX treatment was the wider occurrence of AMC double labelled with anti-BrdU antibody and GSA I-B4 or OX-42 at E16 compared with those in the controls suggesting that the initial increase of GSA I-B4 labelled AMC may be attributed to their proliferation. The drastic reduction of OX-42 and ED1 positive microglial cells notably at E18 may be due to the downregulation of surface antigens as a result of possible suppressive action of dexamethasone. On the basis of present findings, it is concluded that the antigenic expressions of fetal AMC may be modulated by DEX administrated maternally. Such however appeared to be extremely selective as reflected by the varied expression for certain immune molecules at different stages of brain development. This information would be useful in potential use of glucocorticoids in prenatal therapy of brain pathology via maternal circulation.

Intrathecally administered c-fos antisense oligodeoxynucleotide decreases formalin-induced nociceptive behavior in adult rats.

c-fos antisense strategy was applied as a pharmacological approach to characterize its dose-dependent role and reversibility in the reduction of formalin-induced hyperalgesia. Nociceptive behavioral responses (weighted score, flinching response, licking/biting) following formalin (50 microl 5%) injection were assessed in adult Wistar rats receiving different doses (50 nM, 250 nM) of intrathecally administered c-fos antisense oligodeoxynucleotides at different times prior to formalin injections. The treatments dose dependently decreased both Fos immunoreactivity expression in dorsal horn of rat lumbar spinal cord and all nociceptive measures in the tonic phase of the formalin test. c-Fos correlated well with weighted pain score and/or flinching responses, but not with licking/biting behavior. With the exception of a 48-120 h period required for licking/biting behavior to be restored to its normal status, the suppressive effect on c-fos expression and other nociceptive behaviors disappeared 48 h following c-fos antisense oligodeoxynucleotide treatment. The results suggest a pharmacological potential of c-fos antisense oligodeoxynucleotides in the central nervous system to block immediate-early genes and their resulting physiological consequence following noxious stimulus.