Successful Treatment of Herpes Zoster Ophthalmicus Complicated by Intense Orbital Inflammation Using Laser Irradiation over the Stellate Ganglion.

A 56-year-old man presented with right-sided headache and ptosis accompanied by a facial skin rash. He was diagnosed with herpes zoster ophthalmicus (HZO). Despite acyclovir and steroid therapy, the ocular symptoms worsened. Magnetic resonance imaging (MRI) revealed severe orbital inflammation and abnormal lesions in the right trigeminal nucleus and tract. The effects of re-administration of intravenous acyclovir and steroid pulse therapy were limited. Laser irradiation of the stellate ganglion (SGL) and high-dose oral prednisolone therapy were effective. Our experience suggests the efficacy of early multimodal treatment, including SGL, in treating ocular symptoms associated with HZO.

Antepartum ornithine transcarbamylase deficiency.

Ornithine transcarbamylase deficiency (OTCD) is the most common type urea cycle enzyme deficiencies. This syndrome results from a deficiency of the mitochondrial enzyme ornithine transcarbamylase, which catalyzes the conversion of ornithine and carbamoyl phosphate to citrullin. Our case was a 28-year-old female diagnosed with OTCD following neurocognitive deficit during her first pregnancy. Although hyperammonemia was suspected as the cause of the patient's mental changes, there was no evidence of chronic liver disease. Plasma amino acid and urine organic acid analysis revealed OTCD. After combined modality treatment with arginine, sodium benzoate and hemodialysis, the patient's plasma ammonia level stabilized and her mental status returned to normal. At last she recovered without any damage left.

[Two cases of pulmonary embolism in spinal surgeries].

We experienced two cases of pulmonary embolism (PE) in the perioperative period. Although the incidence of perioperative PE is low, it may lead to a critical outcome. The first case is a 59-year-old man without risk factors of PE, scheduled for laminectomy. The end tidal CO2 of 25 mmHg and Pa(CO2) of 48 mmHg developed at the same time during the operation, suggesting PE. He was diagnosed as PE by pulmonary perfusion scan later. The second case was a 71-year-old woman with hypertension and diabetes mellitus, scheduled for laminectomy. Although there were no events during the surgery, she complained of chest pain and dyspnea after the operation. Blood gas analysis showed Pa(O2) of 55 mmHg (FI(O2) 0.4). She was also diagnosed as PE by pulmonary perfusion scan. Both patients made satisfactory progress by appropriate diagnosis and treatment. PE may occur in spite of prevention, and it is important to find out the signs of PE and to prepare for the occurrence of PE.

[Implication of genetic polymorphism on neuropathic pain].

Various conditions are categorized to neuropathic pain, many of which are intractable requiring investigation using multiple approaches. Genetic polymorphism is one of such research fields that have provided much insight into conditions including postherpetic neuralgia, CRPS and postoperative pains. The research in genetic polymorphism is also expected to help expedite realization of the genomic personalized medicine in that selection and dosage of medication are determined according to each patient's specific set of requirements. In addition, the genetic polymorphism research effort in the field of drug discovery in areas such as metabolism, transporters and receptors is also expected to contribute to the clinical practice by expanding our knowledge in each of those areas.

Elevated urinary Cr loss induces a reduction in renal Cr concentration and the negative Cr balance in streptozotocin-induced diabetic mice.

Chromium (Cr) is an essential trace element and is important for normal carbohydrate metabolism, and its deficiency in animals can cause a diabetic-like state. Human and experimental animal studies suggest that urinary Cr excretion is increased in diabetic populations. To investigate whether hyperglycemia-induced elevation of urinary Cr excretion reduces tissue Cr storage conditions, we assessed total Cr balance and Cr distribution in streptozotocin (STZ)-induced diabetic mice. Male C57BL mice were randomly assigned to STZ or control groups and their urine was collected 7, 14, 21 and 28 d after STZ injection. An inductively coupled plasma mass spectrometry instrument equipped with a dynamic reaction cell was used for determination of Cr in urine, plasma and tissues samples. The urinary excretions of Cr were 15.4+/-3.0 and 356+/-62 ng/d, and the renal Cr concentrations were 0.85+/-0.12 and 0.17+/-0.03 ng/mg for the control and diabetes groups, respectively (p<0.01), after 28 d. The Cr balance in STZ-treated mice was distinctly negative due to the increase in urinary Cr loss (p<0.01). These results suggest that in mice, STZ induces a reduction in renal Cr concentration and total negative Cr balance caused by an increase in urinary Cr output.

Major histocompatibility complex haplotype is associated with postherpetic pain in mice.

BACKGROUND: Postherpetic neuralgia is one of the major complications of herpes zoster caused by the reactivation of varicella-zoster virus and is characterized by severe pain. The authors previously showed the association of a human major histocompatibility complex (MHC) haplotype with postherpetic neuralgia. This study was performed to experimentally confirm the role of MHC haplotype in the development of postherpetic pain using a mouse model of postherpetic pain, which corresponds to postherpetic neuralgia. METHODS: BALB/c mice (MHC haplotype: H-2), C57BL/6 mice (MHC haplotype: H-2), and BALB/b mice, a congenic BALB/c strain with H-2, were used. Herpes simplex virus type I was transdermally inoculated on the hind paw. Unilaterally zosteriform skin lesion and pain-related responses (acute herpetic pain) were caused, and some mice showed pain-related responses (postherpetic pain) after the cure of skin lesions. Herpes simplex virus type I antigen and CD3-positive cells were immunostained in the dorsal root ganglion in the acute phase. RESULTS: The incidence (78%) of postherpetic pain in C57BL/6 mice was significantly higher than that (35%) in BALB/c mice (P = 0.004, odds ratio = 6.7). Furthermore, the incidence of postherpetic pain in BALB/b (H-2) was similar to that in C57BL/6. Herpes simplex virus type I antigen-positive cells were less in the dorsal root ganglion of C57BL/6 mice than that of BALB/c mice. CD3-positive T cells were more in the dorsal root ganglion of C57BL/6 mice than BALB/c mice. CONCLUSIONS: These results suggest that the MHC haplotype (H-2) is involved in the incidence of postherpetic pain, and CD3-positive T cells may play a role in its pathogenesis.

Gene expression of beta-catenin is up-regulated in inner dental epithelium and enamel knots during molar tooth morphogenesis in the mouse.

Beta-catenin is a multi-functional molecule that is involved in both cell-cell adhesion and signaling. We analyzed changes in beta-catenin gene expression during mouse molar tooth development by in situ hybridization. Prominent up-regulation of the expression of this gene was evident exclusively in the enamel knot at the early cap stage. During the cap and bell stages, the enamel knot, inner dental epithelium, and differentiating stratum intermedium expressed the beta-catenin gene more strongly than other parts of the enamel organ. During these stages, the strength of the gene expression changed heterogeneously within the inner dental epithelium and stratum intermedium. However, the heterogeneity was not evident at the late bell stage, when the cells in the inner dental epithelium had differentiated into ameloblasts at the cusp tip. No spatiotemporal change in beta-catenin gene expression was apparent in the dental papilla except for the cells that differentiated into odontoblasts, which became negative for the expression of the gene after their differentiation. Thus, the up-regulated expression of the beta-catenin gene was strongly associated with epithelial morphogenesis. These findings raise the possibility that the up-regulation of the gene expression and the stabilization of the protein by Wnt signaling play a role in the regulation of the activities of beta-catenin in tooth morphogenesis.

Expression of glial cell line-derived neurotrophic factor (GDNF) and GDNF family receptor alpha1 in mouse taste bud cells after denervation.

Glial cell line-derived neurotrophic' factor (GDNF) has been isolated as a neurotrophic factor that affects the survival and maintenance of central and peripheral neurons. Using immunocytochemical methods, we examined whether the taste bud cells in mouse circumvallate papillae after transection of the glossopharyngeal nerves expressed GDNF and its receptor, GDNF family receptor alpha1 (GFRalpha1). By 5 and 10 days after denervation, the number of taste buds had decreased markedly; however, the remaining taste bud cells still expressed GDNF and GFRalpha1. By 14 days after denervation, most of the taste buds had disappeared and GDNF- and GFRalpha1-immunoreactive cells were not seen. By 4 weeks after denervation, numerous TrkB-immunoreactive nerve fibers had invaded the papilla and a few taste buds expressing GDNF and GFRalpha1 had regenerated. Thus, GDNF- and GFRalpha1-immunoreactive taste bud cells after denervation vanished following the disappearance of the taste buds and reappeared at the same time as the taste buds reappeared.

Immunohistochemical detection of neurotrophin-3 and -4, and their receptors in mouse taste bud cells.

Neurotrophin-3 (NT3) and neurotrophin-4 (NT4) affect the survival and maintenance of central and peripheral neurons. Using an immunohistochemical method, we examined whether the taste bud cells in the circumvallate papillae of normal mice expressed NT3, NT4, and their respective receptors TrkC and TrkB, and if so, what type of cells in the taste buds expressed them. Double immunostaining for either of them and PGP 9.5, NCAM, or gustducin was used to determine which cell types expressed which neurotrophins and receptors. Normal taste bud cells expressed NT3, NT4, and the TrkB receptor, but not TrkC. The percentage of NT3-immunoreactive cells among all taste bud cells was 89.0%, that of NT4-immunoreactive cells, 58.6%, and that of TrkB-immunoreactive cells, 80.8%. Almost none of the NT4-immunoreactive cells were reactive with anti-PGP 9.5 or the anti-NCAM antibody, but they could be stained with anti-gustducin, revealing that NT4-immunoreactive cells were contained only in the type-II--and possibly type-I--cell population. On the other hand, NT3-, and TrkB-immunoreactive cells included type-III cells, together with type-II, -I, and basal cells, because they were positive for PGP 9.5 and gustducin. We conclude that NT4 may exert trophic actions on all types of taste bud cells by binding to their TrkB receptors, and NT3 may also have a similar, though negligible role.

Distinct expression pattern of insulin-like growth factor family in rodent taste buds.

The insulin-like growth factor (IGF) system is an important regulator of growth and differentiation in a variety of tissues. In the present study, the expression of IGF family members in the taste buds of mice and rats was examined. By reverse transcriptase polymerase chain reaction (RT-PCR) analysis, mRNA of IGF-I and -II, IGF-I receptor (IGF-IR), insulin receptor (insulin R), and IGF-binding protein (IGFBP)-2, -3, -4, -5, and -6 was detected in the taste bud-containing epithelium of the circumvallate papillae of mice. As suggested by the study using degenerate PCR (McLaughlin [2000] J. Neurosci. 20:5679-5688), IGF-IR was expressed in most of the taste bud cells of adult mice, as found by immunohistochemistry, and in those of postnatal day (P) 6 mice by in situ hybridization. Insulin R, which has strong homology to IGF-IR, was also detected in most of the taste bud cells of mice by immunohistochemistry and in situ hybridization. IGF-I immunoreactivity was detected in a few taste bud cells and in the epithelium surrounding taste buds. Northern blot analysis revealed that the amount of IGF-I mRNA in taste bud-containing epithelium was very low compared with that in liver. IGF-II immunoreactivity was weakly detected in mouse taste buds and the surrounding epithelium. In the rat tissue, a subset of the taste bud cells was positive for IGF-II. Among the six IGFBPs, IGFBP-2, -5, and -6 were detected in the mouse taste buds: IGFBP-2 and -5 immunoreactivity was seen in the majority of the taste bud cells, whereas IGFBP-6 immunoreactivity was found in the nerve fibers innervating the taste buds. In situ hybridization study also revealed that IGFBP-2 and -5 mRNA was synthesized in the taste buds of P6 mice and that the expression of these mRNAs overlapped in von Ebner's glands. These data reveal that IGF-I and -II might be produced in taste bud cells and (or) surrounding lingual epithelium and act through IGF-IR and insulin R locally in a paracrine and autocrine manner. The activity of these IGFs may be modulated through their interaction with IGFBP-2, -5, and 6.

Expression of GDNF and GFR alpha 1 in mouse taste bud cells.

GDNF (glial cell line-derived neurotrophic factor) affects the survival and maintenance of central and peripheral neurons. Using an immunocytochemical method, we examined whether the taste bud cells in the circumvallate papillae of normal mice expressed GDNF and its GFR alpha 1 receptor. Using double immunostaining for either of them and NCAM, PGP 9.5, or alpha-gustducin, we additionally sought to determine what type of taste bud cells expressed GDNF or GFR alpha 1, because NCAM is reported to be expressed in type-III cells, PGP 9.5, in type-III and some type-II cells, and alpha-gustducin, in some type-II cells. Normal taste bud cells expressed both GDNF and GFR alpha 1. The percentage of GDNF-immunoreactive cells among all taste bud cells was 31.63%, and that of GFR alpha 1-immunoreactive cells, 83.21%. Confocal laser scanning microscopic observations after double immunostaining showed that almost none of the GDNF-immunoreactive cells in the taste buds were reactive with anti-NCAM or anti-PGP 9.5 antibody, but could be stained with anti-alpha-gustducin antibody. On the other hand, almost all anti-PGP 9.5- or anti-alpha-gustducin-immunoreactive cells were positive for GFR alpha 1. Thus, GDNF-immunoreactive cells did not include type-III cells, but type-II cells, which are alpha-gustducin-immunoreactive; on the other hand, GFR alpha 1-immunoreactive cells included type-II and -III cells, and perhaps type-I cells. We conclude that GDNF in the type-II cells may exert trophic actions on type-I, -II, and -III taste bud cells by binding to their GFR alpha 1 receptors.

The human histocompatibility leukocyte antigen (HLA) haplotype is associated with the onset of postherpetic neuralgia after herpes zoster.

In some herpes zoster patients, pain persists for more than 3 months or more after healing of vesicular eruptions; this condition is termed postherpetic neuralgia (PHN). We have recently reported the association of the human histocompatibility leukocyte antigens (HLA) haplotype, HLA-A*3303-B*4403-DRB1*1302 with PHN patients; however, it has not been determined whether the haplotype is also associated with herpes zoster that did not develop subsequent PHN. To distinguish whether the haplotype is associated with herpes zoster or the development of PHN, we examined if herpes zoster patients without subsequently PHN are also associated with the HLA haplotype or not. Herpes zoster patients were followed up for more than 6 months, and HLA alleles and haplotypes were compared among the PHN patients (n = 52) the herpes zoster patients who did not develop PHN (n = 42) and healthy controls (n = 125). The frequencies of the risk haplotype in the PHN patients, in the healthy controls and in the herpes zoster patients without subsequent PHN were 16.3, 5.2 and 4.8%, respectively. While the frequency of the risk haplotype was significantly higher in the PHN patients than in the healthy controls (P = 0.0006) no difference was observed between the herpes zoster patients without subsequent PHN and the healthy controls. No significant association was found between the duration of symptoms or the site of herpes zoster and the HLA alleles and the haplotype. These results suggest that the HLA-A*3303-B*4403-DRB1*1302 haplotype plays an important role in the development of PHN after herpes zoster, but not in the onset of herpes zoster.

Expression of bHLH transcription factors NSCL1 and NSCL2 in the mouse olfactory system.

We examined the expression of basic helix-loop-helix transcription factors NSCL1 and NSCL2 in the olfactory epithelium (OE) and the vomeronasal organ (VNO) during development. As detected by in situ hybridization, at embryonic day (E) 10 NSCL1 was weakly expressed in the entire olfactory placodes. From E12 to postnatal day (P) 3, NSCL1 was expressed in olfactory receptor neurons (ORNs) and receptor neurons of the VNO. The expression pattern of NSCL2 was similar to that of NSCL1. By Northern blot analysis, strong expression of NSCL1 was detected in the OE from E12 to P7, but the expression there was low in the adult (P35). NSCL2 mRNA was detected in the E12 and P1 OE, but its level was very low in the P7 and adult OE. The spatial pattern of expression suggests that NSCL1 and NSCL2 contribute to the maturation of ORNs (VNO receptor neurons) or maintenance of their differentiated state. Moreover, the temporal pattern of expression suggests that NSCL1 and NSCL2 may function during development rather than in the adult stage.

Expression of BDNF and TrkB in mouse taste buds after denervation and in circumvallate papillae during development.

BDNF (brain-derived neurotrophic factor) is a member of the neurotrophin family which affects the proliferation and survival of neurons. Using an immunocytochemical method, we examined the expression of BDNF and its receptor, TrkB, in the taste bud cells of the circumvallate papillae of normal mice and of mice after transection of the glossopharyngeal nerves. We additionally observed the expression of BDNF and TrkB in the developing circumvallate papillae of late prenatal and early postnatal mice. In normal untreated mice, BDNF was expressed in most of the taste bud cells; TrkB was detected in the plasma membrane of taste bud cells and in the nerve fibers. Double-labeling studies showed that BDNF and NCAM (neural cell adhesion molecule) or TrkB and NCAM colocalized in some of the taste bud cells, but that most taste bud cells were immunopositive for only BDNF or TrkB. NCAM-immunoreactive cells are known to be type-III cells, which have afferent synaptic contacts with the nerve terminals. Five days after denervation, the number of taste buds and nerve fibers markedly decreased; however, the remaining taste bud cells still expressed BDNF and TrkB. By 10 days after denervation, most of the taste buds had disappeared, and there were a few TrkB-immunoreactive nerve fibers in the connective tissue core. By 4 weeks after denervation, numerous TrkB-immunoreactive nerve fibers had invaded the papillae, and a few taste buds expressing BDNF and TrkB had regenerated. At E (embryonic day) 15 during development, the circumvallate papillae appeared, and then TrkB-immunoreactive nerve fibers entered the connective tissue core, and some of these fibers further invaded among the dorsal epithelial cells of the papillae. TrkB-immunoreactive oval-shaped cells were occasionally found in the dorsal epithelium. Such TrkB-immunoreactive nerve fibers and cells were also observed at E16-18. However, BDNF was not expressed in the papillae through the late prenatal days of E15 to E18. At P (postnatal day) 0, a cluster of BDNF-and TrkB-immunoreactive cells appeared in the dorsal epithelium of the papillae, and was presumed to be primitive taste buds. We conclude that TrkB-immunoreactive nerve fibers are necessary for papillary and taste bud formation during development and for the regeneration of taste buds after denervation. BDNF in the taste bud cells may act as a neurotrophic factor for innervating sensory neurons--through TrkB receptors of the axons of those neurons, and also may exert autocrine and paracrine trophic actions on neighboring taste bud cells by binding to their TrkB receptors.

Expression of Hes6 and NeuroD in the olfactory epithelium, vomeronasal organ and non-sensory patches.

Basic helix-loop-helix transcription factors NeuroD and Hes6 promote neuronal differentiation. The expression of their genes in the olfactory epithelium (OE), vomeronasal organ (VNO) and the non-sensory patches of the posterior nasal cavity of mice was examined. As detected by in situ hybridization, Hes6 was expressed in a basal progenitor layer of the embryonic OE. After birth, the expression of Hes6 was detected in a cell layer above the basement membrane, globose basal cells (GBCs). Expression of NeuroD in the embryonic OE was in agreement with that previously described; and in the postnatal OE, it was detected in cells of GBC layer and cells upper to GBCs. In the VNO, Hes6 was expressed throughout the sensory epithelium (S-VNO) at embryonic day 12, and later became restricted to a single layer of cells in the basal region of the S-VNO, where Hes5-expressing undifferentiated cells were present. NeuroD was expressed throughout the S-VNO during the embryonic stage. After birth, Hes6 and NeuroD expressions were observed in the border between the S-VNO and non-sensory VNO. Immunohistochemistry using anti-NeuroD antibody revealed that NeuroD-positive cells were still present not only at the edges but also in the center of the S-VNO until P3. These findings suggest that Hes6 and NeuroD are expressed in progenitors of chemoreceptor neurons and that the expression of Hes6 precedes that of NEUROD: Moreover, in the regenerating VNO of bulbectomized mice, NeuroD-positive cells were observed both at the edges and in the center of the S-VNO, suggesting that neuronal turnover occurred in both regions. Moreover, in the dorsal fossa of the posterior nasal cavity, several non-sensory patches are formed between postnatal (P) days 10 and 21 because of programmed death of ORNs and GBCs. During embryonic stages, the expression of Hes6 and NeuroD in the OE showed no regional differences. At P3-P7, expression of NeuroD and Hes6 disappeared in the region corresponding to the presumptive non-sensory patches. The loss of these genes may stop the differentiation and may cause apoptosis of GBCs and ORNs.

Expression of neural cell-adhesion molecule mRNA during mouse molar tooth development.

This study employed in situ hybridisation using a probe recognising all isoforms of the molecule. Expression of the molecule in tooth germs started at embryonic day 13, when they were at the bud stage. Both inner cells of the epithelial bud and peripheral cells of the dental mesenchyme were positive. At the cap stage, positive cells were found in the inner part of the enamel organ but only in a limited area near the outer enamel epithelium. In the mesenchyme at the cap stage, expression was weak in the dental papilla and strong in the follicle. From the bell stage onward, epithelial cells in the enamel organ were negative except for the cells of the stratum intermedium, which were transiently positive at early and late bell stages. In the dental papilla, expression had mostly ceased during and after the bell stage, although transient expression was found in cuspal areas at the early bell stage. The dental follicle strongly expressed neural cell-adhesion molecule (NCAM) to the end of the experimental period, at post-natal day 4. In contrast to the first molar at its earliest stage of appearance, in which both the thickened epithelium and surrounding mesenchyme were negative for the expression of the molecule, the second molar appeared as a combination of extending epithelial thickenings and mesenchymal cells strongly positive for its expression. This study newly identifies the dental papilla and the stratum intermedium as NCAM-expressing sites.

Expression of insulin-like growth factor family in the rat olfactory epithelium.

The goal of this study was to determine the cellular sites of insulin-like growth factor (IGF) family expression in the rat olfactory epithelium. By RT-PCR analysis, mRNAs of IGF-I, II, IGF-I receptor (IGF-IR), and IGF binding proteins (IGFBPs) 2, 3, 4, 5, and 6 were found to be expressed in the olfactory mucosa. Immunoreactivity for IGF-IR was restricted to a subset of olfactory receptor cells whose cell bodies were situated in the basal region of the olfactory epithelium. Intense IGF-I immunoreactivity was detected in the supporting cells, whereas IGF-II immunoreactivity was observed in the lamina propria, but not in the epithelium. Immunoreactivities for IGFBP-2, IGFBP-3, and IGFBP-6 were detected in olfactory receptor cells. In addition, axon bundles in the lamina propria displayed an intense reaction for IGFBP-6. IGFBP-4 immunoreactivity was restricted to the apex of the olfactory epithelium. Intense IGFBP-5 immunoreactivity was observed in Bowman's glands. These results suggest that IGF-I is secreted from supporting cells and affects its receptor- expressing olfactory cells. IGFBPs may modulate IGF-I activity via their production by Bowman's glands, olfactory cells, and supporting cells themselves.

Expression of NeuroD in the mouse taste buds.

NeuroD, a basic helix-loop-helix transcription factor, has been shown to play a role in the differentiation of neurons, olfactory cells, and neuroendocrine tissues. Since the taste buds have characteristics of typical paraneurons, we examined the expression of NeuroD in the taste buds of mice. By RT-PCR analysis, NeuroD mRNA was found to be expressed in the epithelium of circumvallate papillae-containing taste buds, but not in the lingual epithelium lacking them. NeuroD immunoreactivity was detected in a subset of taste bud cells in the circumvallate, foliate, and fungiform papillae and in the soft palate. NeuroD-expressing cells had a spindle-like shape, first appeared at postnatal day 3, and increased in number during postnatal development. After bisection of the glossopharyngeal nerves, NeuroD-expressing cells decreased in number at day 4 and disappeared from the trench wall of the circumvallate papillae by day 14. A few NeuroD-expressing taste buds reappeared at postoperative day 28. Denervation and regeneration experiments showed that expression of NeuroD in the taste buds was dependent upon gustatory innervation. Double immunolabeling with gustducin or with neural cell adhesion molecule (NCAM) showed that NeuroD-expressing cells did not express NCAM, but did express gustducin. These results suggest that NeuroD is expressed in a mature cell type, type-II cells, but not in type-III cells.