Changes in the expression of α-tocopherol-related genes in liver and mammary gland biopsy specimens of peripartum dairy cows.

Blood α-tocopherol (α-Toc) concentrations decline gradually throughout the prepartum period, reaching the nadir after calving in dairy cows. The 6 α-Toc-related molecules [α-Toc transfer protein (TTPA); afamin; scavenger receptor class B, Type I; ATP-binding cassette transporter A1; tocopherol-associated protein (SEC14L2); and cytochrome P450 family 4, subfamily F, polypeptide 2 (CYP4F2)] are expressed in liver and other peripheral tissues. These molecules could regulate α-Toc transport, blood concentrations, and metabolism of α-Toc. Therefore, the aim of this study was to evaluate the changes in the expression of α-Toc-related genes in liver and mammary gland tissues of dairy cows around calving, which have remained elusive until now. In experiment (Exp.) 1, 28 multiparous Holstein cows were used (from -5 to 6 wk relative to parturition) to monitor the changes in dietary α-Toc intake, blood concentrations of α-Toc, and lipoproteins; in Exp. 2, 7 peripartum Holstein cows were used (from -4 to 4 wk relative to parturition) for liver tissue biopsy; and in Exp. 3, 10 peripartum Holstein cows were used (from -8 to 6 wk relative to parturition) to carry out the mammary gland tissue biopsy and milk sampling. In Exp. 1, the serum α-Toc concentrations declined gradually with decreasing amount of α-Toc intake and plasma high-density lipoprotein concentrations toward calving time. However, in the early lactation period after calving, serum α-Toc concentrations remained at a lower concentration despite the recovery of α-Toc intake and plasma high-density lipoprotein concentrations. In Exp. 2, just after calving, the TTPA, SEC14L2, afamin, and albumin mRNA expression levels in the liver were temporarily downregulated, and the hepatic mRNA levels of endoplasmic reticulum stress-induced unfolded protein response markers and acute-phase response marker increased at calving. In Exp. 3, the concentrations of α-Toc in colostrum were greater than those in precolostrum (samples were collected at wk -1 relative to parturition) and mature milk. The expression of TTPA, SEC14L2, and CYP4F2 mRNA in bovine mammary gland tissue was detected. However, TTPA and SEC14L2 mRNA expressions showed the opposite trends: the expression levels of TTPA mRNA peaked whereas SEC14L2 mRNA reached a nadir at calving. These results indicate that the expression of α-Toc-related genes involved in specific α-Toc transfer and metabolism in the liver and mammary gland are altered during calving. Moreover, these changes might be associated with the maintenance of lower serum α-Toc concentrations after calving.

Postweaning changes in the expression of chemerin and its receptors in calves are associated with the modification of glucose metabolism.

Chemerin, originally known as a chemoattractant derived from adipose tissue and the liver, has been reported to have regulatory functions in gluconeogenesis, peripheral insulin sensitivity, and insulin secretion. This study was conducted to assess the postweaning changes in expression of this cytokine and its physiological role in the modification of glucose metabolism associated with weaning. Eighteen tissue samples were collected from Holstein calves (90 d of age; n = 4) to investigate the tissue distributions of chemerin and its receptors genes. was highly expressed in the liver, and secreted chemerin protein was found in the plasma. Among the receptors of chemerin, and were ubiquitously expressed whereas was predominantly expressed in the liver. The changes in glucose metabolism and expression of these genes after weaning were assessed by comparing suckling calves (n = 6) and weaned calves (n = 8) of Japanese Black cattle. No significant difference was observed in plasma glucose levels between suckling and weaned calves (P = 0.22), whereas the plasma level of total ketone bodies was significantly higher in weaned calves (P < 0.01). Plasma levels of insulin and cortisol did not differ between suckling and weaned calves. The mRNA levels of certain key enzymes involved in hepatic gluconeogenesis were also altered; for instance, level was lower in postweaning calves (P < 0.05) and () level tended to be higher after weaning (P = 0.08). However, was not altered after weaning. The plasma levels of hepatic stress indicators were also changed, with aspartate transaminase, alanine transaminase, and lactate dehydrogenase being significantly elevated in postweaning calves (P < 0.05). Chemerin protein in liver tissue was less abundant in weaned calves (P < 0.05), although there were no changes in its transcript levels. The abundance of plasma chemerin protein did not change after weaning (P = 0.95). In summary, these data indicate that as a consequence of weaning, which causes physiological stress and alters hepatic metabolism, chemerin protein expression within the liver is downregulated, indicating that chemerin plays a role in the upregulation of hepatic expression via its inhibitory effect on hepatic gluconeogenesis.

Expression of α-tocopherol-associated genes and α-tocopherol accumulation in Japanese Black (Wagyu) calves with and without α-tocopherol supplementation.

The aim of the study was to clarify 1) the distribution of 6 α-tocopherol (α-Toc)-associated gene expressions in 20 major tissues, including metabolic, reproductive, endocrine, immune, and digestive and absorptive tissues, in relation to α-Toc status and 2) the change in expression patterns of the genes induced when α-Toc was orally administered to Japanese Black (JB) calves. This study examined weaned male JB calves ( = 10), of which 5 calves were orally administered α-Toc for 2 wk (30 IU·kg BW·d; TOC group). The others did not receive the α-Toc supplement and were the control (CONT) group. The 20 tissues and venous blood (serum) were sampled on the final day. In both groups, the mean mRNA expression levels for α-Toc transfer protein, afamin (AFM), ATP-binding cassette transporter A1, and tocopherol-associated protein were greatest in the liver ( < 0.05), whereas scavenger receptor class B, Type I (SR-BI) mRNA was greatest in the adrenal gland ( < 0.05). The gene for cytochrome P450 family 4, subfamily F, polypeptide 2 was most highly expressed in the liver, testes, and adrenal gland. The α-Toc content was greatest ( < 0.05) in the testes of the 20 sampled tissues in the CONT group. However, the levels in the testes and jejunum were similar and greater ( < 0.05) than the levels in the other 18 tissues in the TOC group. The mean increase in α-Toc levels after oral α-Toc administration (mean α-Toc content for the TOC group divided by the CONT group content) were greater ( < 0.05) in the jejunum (40.7-fold) and duodenum and liver (26.3- and 23.1-fold) than in the serum (7.8-fold). In the liver, α-Toc administration significantly increased ( < 0.05) the AFM and SR-BI mRNA expression levels. The results show that the liver may play an important role in the regulation of α-Toc disposition, but other peripheral tissues that accumulate large amounts of α-Toc could moderate the local α-Toc status and functions, as inferred from the high expressions of the α-Toc-associated genes in JB calves.

Efficiency of dual-cured resin cement polymerization induced by high-intensity LED curing units through ceramic material.

OBJECTIVE: This study aimed to evaluate the ability of high-intensity light-emitting diode (LED) and other curing units to cure dual-cured resin cement through ceramic material. METHODS: A halogen curing unit (Jetlite 3000, Morita), a second-generation LED curing unit (Demi, Kerr), and two high-intensity LED curing units (PenCure 2000, Morita; Valo, Ultradent) were tested. Feldspathic ceramic plates (VITABLOCS Mark II, A3; Vita Zahnfabrik) with thicknesses of 1.0, 2.0, and 3.0 mm were prepared. Dual-cured resin cement samples (Clearfil Esthetic Cement, Kuraray Noritake Dental) were irradiated directly or through one of the ceramic plates for different periods (5, 10, 15, or 20 seconds for the high-intensity LED units and 20, 40, 60, or 80 seconds for the others). The Knoop hardness test was used to determine the level of photopolymerization that had been induced in the resin cement. Data were analyzed by one-way analysis of variance and Dunnett's post-hoc test to identify test-control (maximum irradiation without a ceramic plate) differences for each curing unit (p<0.05). RESULTS: For all curing units, the curing conditions had a statistically significant effect on the Knoop hardness numbers (KHNs) of the irradiated cement samples (p<0.001). In general, the KHN decreased with increasing plate thickness and increased as the irradiation period was extended. Jetlite 3000 achieved control-level KHN values only when the plate thickness was 1.0 mm. At a plate thickness ≥2.0 mm, the LED units (except for PenCure 2000 at 3.0 mm) were able to achieve control-level KHN values when the irradiation time was extended. At a plate thickness of 3.0 mm, irradiation for 20 seconds with the Valo or for 80 seconds with the Demi were the only methods that produced KHN values equivalent to those produced by direct irradiation. CONCLUSION: Regardless of the type of curing unit used, indirect irradiation of dual-cured resin cement through a ceramic plate resulted in decreased KHN values compared with direct irradiation. When the irradiation period was extended, only the LED units were able to achieve similar KHN values to those observed under direct irradiation in the presence of plates ≥2.0-mm thick. High-intensity LED units require a shorter irradiation period than halogen and second-generation LED curing units to obtain KHN values similar to those observed during direct irradiation.

Hypothalamic ATF3 is involved in regulating glucose and energy metabolism in mice.

AIMS/HYPOTHESIS: The pancreas and hypothalamus are critical for maintaining nutrient and energy homeostasis, and combined disorders in these organs account for the onset of the metabolic syndrome. Activating transcription factor 3 (ATF3) is an adaptive response transcription factor. The physiological role of ATF3 in the pancreas has been controversial, and its role in the hypothalamus remains unknown. To elucidate the roles of ATF3 in these organs, we generated pancreas- and hypothalamus-specific Atf3 knockout (PHT-Atf3-KO) mice in this study. METHODS: We crossed mice bearing floxed Atf3 alleles with Pdx1-cre mice, in which cre is specifically expressed in the pancreas and hypothalamus, and analysed metabolic variables, pancreatic morphology, food intake, energy expenditure and sympathetic activity in adipose tissue. We also used a hypothalamic cell line to investigate the molecular mechanism by which ATF3 regulates transcription of the gene encoding agouti-related protein (Agrp). RESULTS: Although PHT-Atf3-KO mice displayed better glucose tolerance, neither plasma glucagon nor insulin level was altered in these mice. However, these mice exhibited higher insulin sensitivity, which was accompanied by a leaner phenotype due to decreased food intake and increased energy expenditure. We also observed decreased hypothalamic Agrp expression in PHT-Atf3-KO mice. Importantly, an increase in ATF3 levels is induced by fasting or low glucose in the hypothalamus. We also showed that ATF3 interacts with forkhead box-containing protein, O subfamily 1 (FoxO1) on the Agrp promoter and activates Agrp transcription. CONCLUSIONS/INTERPRETATION: Our results suggest that ATF3 plays an important role in the control of glucose and energy metabolism by regulating Agrp.

Key role of ATF3 in p53-dependent DR5 induction upon DNA damage of human colon cancer cells.

Stress response gene ATF3 is one of the p53 target genes and has a tumor suppressor role in cancer. However, the biological role of p53-ATF3 pathway is not well understood. Death receptor 5 (DR5) is a death domain-containing transmembrane receptor that triggers cell death upon binding to its ligand TRAIL (tumor necrosis factor-related apoptosis-inducing ligand), and a combination of TRAIL and agents that increase the expression of DR5 is expected as a novel anticancer therapy. In this report, we demonstrate that ATF3 is required for efficient DR5 induction upon DNA damage by camptothecin (CPT) in colorectal cancer cells. In the absence of ATF3, induction of DR5 messenger RNA and protein is remarkably abrogated, and this is associated with reduced cell death by TRAIL and CPT. By contrast, exogenous expression of ATF3 causes more rapid and elevated expression of DR5, resulting in enhanced sensitivity to apoptotic cell death by TRAIL/CPT. Reporter assay and DNA affinity precipitation assay demonstrate that at least three ATF/CRE motifs at the proximal promoter of the human DR5 gene are involved in the activation of DNA damage-induced DR5 gene transcription. Furthermore, ATF3 is shown to interact with p53 to form a complex on the DR5 gene by Re-chromatin immunoprecipitation assay. Taken together, our results provide a novel insight into the role of ATF3 as an essential co-transcription factor for p53 upon DNA damage, and this may represent a useful biomarker for TRAIL-based anticancer therapy.

Role of Necl-5 in the pathophysiology of colorectal lesions induced by dimethylhydrazine and/or dextran sodium sulphate.

Necl-5 is an immunoglobulin-like molecule that was originally identified as a poliovirus receptor. Although Necl-5 expression is often up-regulated in cancer cells, its pathophysiological significance in the development of cancer remains unclear. We investigated the roles of Necl-5 in the development of colitis-associated neoplasia. Necl-5-deficient mice were generated and treated with dimethylhydrazine (DMH) and/or dextran sodium sulphate (DSS) to induce colitis and its associated neoplasias. Colon tissues were examined for histology, Ki-67 expression by immunohistochemistry and K-ras gene mutation. Colon tumours occurred significantly less frequently in heterozygous (Necl-5(+/-)) or homozygous Necl-5-deficient (Necl-5(-/-)) mice than in wild-type (WT) mice with DMH/DSS treatment. Total ulcer index and inflammatory cell infiltration were significantly lower in Necl-5(-/-) mice than in WT mice with DSS alone or DMH/DSS treatment. Colon tumours in both WT and Necl-5(-/-) mice showed high cell proliferation ability but lacked K-ras mutation. The total Ki-67 labelling index in non-neoplastic colon epithelium was significantly higher in WT (45.9 +/- 0.94) than in Necl-5(+/-) (34.3 +/- 1.40) or Necl-5(-/-) (27.7 +/- 1.15) mice with DMH/DSS treatment (p < 0.001). Necl-5 plays a role in the development of colitis-associated cancer by up-regulating colonic mucosal cell proliferation.

Chitosan-containing gum chewing accelerates antibacterial effect with an increase in salivary secretion.

OBJECTIVES: This study was designed to confirm the mechanical efficacy of chewing chitosan-containing gum to suppress the growth of oral bacteria compared to a mouth rinse, and to demonstrate the increased salivary secretion due to chewing chitosan-containing gum. METHOD: Twelve healthy subjects were recruited from among the staff and students of Nagasaki University School of Dentistry. For the slab of gum study, the subjects chewed chitosan-containing gum for 5 min and then rested for 5 min. For the testing of the chitosan mouth rinse solution, the subjects gargled 10 mL of solution for 30s followed by resting for 9 min 30s. These protocols were continuously repeated five times for 50 min on the same day. For the salivary secretion experiment, the gum chewing was repeated three times per day for 2 days. RESULTS: The amount of oral bacteria in the subjects who chewed chitosan-containing gum significantly decreased during all three sampling times except at 60 min for total bacteria in comparison to those in the rinse group. Chitosan-containing gum chewing also significantly increased the secretion of saliva. CONCLUSIONS: Chitosan-containing gum chewing has a greater antibacterial effect and it also increases salivary secretion. The present findings strongly indicate that the application of natural materials such as chitosan is useful for both oral health and the quality of life.

Can low frequency sound stimulation during posturography help diagnosing possible perilymphatic fistula in patients with sensorineural hearing loss and/or vertigo?

Perilymphatic fistula (PLF) is often difficult to diagnose because of the similar symptomatology, such as vertigo, tinnitus and hearing loss, which is found in several inner ear diseases. We attempted to correlate a positive result of low frequency sound (LFS) stimulation tests in posturography with the presence or absence of a PLF confirmed by transtympanic endoscopy in 209 patients with various inner ear diseases (Meniere's disease ( n=128), vestibulopathy ( n=41), cochleopathy ( n=28) and sudden deafness ( n=12). LFS provoked unsteadiness in posturography without PLF in 24 patients with Meniere's disease, in 5 patients with vestibulopathy, in 3 patients with cochleopathy and in 2 patients with sudden deafness. In one patient, tympanoscopy revealed fistula in the round window membrane that was covered with a fibrinous layer. In four cases there was abnormal light reflex in the round window but without PLF. In eight cases, Hennebert's sign was present with nystagmus, without PLF. We conclude that pathological responses to the LFS test in posturography can also be encountered in other inner ear diseases without PLF.

Role of transtympanic endoscopy of the middle ear in the diagnosis of perilymphatic fistula in patients with sensorineural hearing loss or vertigo.

BACKGROUND: The diagnosis of perilymphatic fistula (PLF) is often difficult, and therefore the condition can be overlooked. Tympanoscopy presents an alternative procedure for visualising the middle ear anatomy, and it may help to diagnose PLF. AIM: The aim of this study was to evaluate the use of middle ear endoscopy in establishing the diagnosis of PLF and in defining its incidence in patients with sensorineural hearing loss and/or vertigo and tinnitus. SUBJECTS AND METHODS: Two hundred and sixty-five patients (22-80 years of age, mean 48 years) were prospectively and consecutively referred for middle ear examination with tympanoscopy. Tympanoscopy was performed using endoscopes with visual angles of 5 and 25 degrees and an outer diameter of 1.7 mm. The round window niche (with its secondary membrane), the oval window with a stapes superstructure, a part of the facial recess and the area in the fissula ante fenestram were examined and video-recorded. RESULTS: For 1 patient, tympanoscopy revealed fistula in the round window membrane that was covered with a fibrinous layer. In 4 cases abnormal mucosal shining appeared in the round window, but no PLF was present. In 7 cases the tympanic cavity could not be visualised because of the adhesive tympanic membrane, abnormal anatomy or the prominent exostoses of the external ear canal. In 6 cases a postendoscopic middle ear infection was found. No permanent tympanic membrane perforation occurred in any of the patients in this study. CONCLUSIONS: Tympanoscopy is a rapid examination tool with which to verify certain areas of the middle ear anatomy, but it is of limited value for ruling out the presence of PLF.

Cochlear blood flow measurement in patients with Ménière's disease and other inner ear disorders.

To identify an eventual vascular aetiology in different diseases with sensorineural hearing loss (SNHL), the cochlear blood flow (CoBF) was measured using laser Doppler flowmetry (LDF) in 69 patients with Ménière's disease (MD), 38 patients with progressive SNHL and 8 patients with sudden deafness. The mean CoBF amplitude at rest was 0.70 mV (SD 0.25) for patients with MD, 0.66 mV (SD 0.21) for patients with SNHL and 0.69 mV (SD 0.23) for those with sudden deafness. No statistically significant difference was observed between the groups with respect to the CoBF amplitudes at rest or during the Valsalva manoeuvre. There was a statistically significant correlation (r = -0.4, p < 0.05) between the hearing level and CoBF amplitude in the SNHL group only. It is concluded that the reduction in vascular flow may not be the main aetiological factor in Ménière's disease or sudden deafness. In SNHL the correlation of hearing level with the level of the LDF amplitude can be secondary to the progression of hearing loss.

Basic fibroblast growth factor in nasal polyps immunohistochemical and quantitative findings.

Basic fibroblast growth factor (bFGF) is a mediator with potent mitogenic properties. Increased amounts of this mediator have been demonstrated in damaged lung tissue, and it has been suggested to increase the healing of gastro-duodenal ulcers. In order to quantify the amounts and document the localization of bFGF in nasal polyps, polyp tissue from 12 patients undergoing polypectomy was analyzed by ELISA and immunohistochemistry. Mucosa from the inferior turbinate was analyzed in the same manner for comparison. The amount of bFGF detected in polyp tissue was significantly higher than that in turbinate mucosa. The amount of bFGF was also significantly higher in the group of patients with high degree of inflammation. The immunohistochemical findings demonstrated abundant bFGF activity mainly in the glandular acini, in the epithelium, in infiltrating inflammatory cells and in the vessel walls. We propose that bFGF may contribute in a significant way to the formation of nasal polyps.

Molecular epidemiology of Arthroderma benhamiae, an emerging pathogen of dermatophytoses in Japan, by polymorphisms of the non-transcribed spacer region of the ribosomal DNA.

In Japan, several isolates of Arthroderma benhamiae, a teleomorphic member of Trichophyton mentagrophytes complex, which were not found by earlier mating studies, have recently been recovered from human and animal dermatophytoses. In the present study, intraspecies polymorphism of A. benhamiae isolated in Japan was investigated using restriction fragment length polymorphisms (RFLP) of the non-transcribed spacer (NTS) region of the ribosomal DNA (rDNA), a method introduced to detect intraspecies polymorphisms of other dermatophyte species, such as T. rubrum. Based on their restriction profiles, there were five DNA types out of eight strains of A. benhamiae isolated in Japan. None of the five DNA types were found among the registered tester strains of A. benhamiae. Therefore, several different strains of A. benhamiae may have been brought into Japan separately.

Role of Rab3 GDP/GTP exchange protein in synaptic vesicle trafficking at the mouse neuromuscular junction.

The Rab3 small G protein family consists of four members, Rab3A, -3B, -3C, and -3D. Of these members, Rab3A regulates Ca(2+)-dependent neurotransmitter release. These small G proteins are activated by Rab3 GDP/GTP exchange protein (Rab3 GEP). To determine the function of Rab3 GEP during neurotransmitter release, we have knocked out Rab3 GEP in mice. Rab3 GEP-/- mice developed normally but died immediately after birth. Embryos at E18.5 showed no evoked action potentials of the diaphragm and gastrocnemius muscles in response to electrical stimulation of the phrenic and sciatic nerves, respectively. In contrast, axonal conduction of the spinal cord and the phrenic nerve was not impaired. Total numbers of synaptic vesicles, especially those docked at the presynaptic plasma membrane, were reduced at the neuromuscular junction approximately 10-fold compared with controls, whereas postsynaptic structures and functions appeared normal. Thus, Rab3 GEP is essential for neurotransmitter release and probably for formation and trafficking of the synaptic vesicles.

[Molecular epidemiology of Sporothrix schenckii].

Restriction fragment length polymorphism (RFLP) in mitochondrial DNA (mtDNA) of clinical and environmental isolates of Sporothrix schenckii was investigated. Among mtDNA RFLP patterns with Hae III, of 14 environmental isolates morphologically identified as S. schenckii, only 2 isolates were confirmed as S. schenckii, while of more than 500 clinical isolates, all were confirmed to have this condition. Therefore, RFLP analysis of mtDNA is essential for the identification of environmental, but not clinical, isolates of S. schenckii. Isolates of Sporothrix schenckii were classified into 23 mtDNA types (Types l-23) based on mtDNA RFLP patterns with HaeIII and clustered into two major groups by phylogeny, Group A (Types 1-3, 11, 14-19, 22 and 23) and Group B (Types 4-10, 12, 13, 20 and 21). Group A isolates are predominant in South Africa, North America, Central America and South America, while Group B isolates are predominant in Australia and Japan. In Japan, the relative distribution of the mtDNA types varied with geographic region: Types 4, 6 are comparatively abundant in West Japan (Kansai and Kyushu districts), Type 5 is comparatively abundant in East Japan (Tokai, Kanto and Tohoku districts) and Type 2 is abundant in the Hokuriku district. Type 1 is found only in the Hokuriku district.

Two cases of tinea corporis by infection from a rabbit with Arthroderma benhamiae.

The first cases of tinea corporis with Arthroderma benhamiae in Japan are reported. A 7-year-old girl and a 30-year-old mother in Shimane prefecture suffered from dermatophyte infections on the neck, shoulder, arms and leg. Three isolates from the two patients and a rabbit by which they supposedly were infected, were identified as Trichophyton mentagrophytes. On the bases of mating tests using the tester strains of both the African race and the Americano-European race of A. benhamiae, they were identified as A. benhamiae African race mating type (-). Our results are the first to indicate that both races of A. benhamiae exist in Japan.

Role of rab GDP dissociation inhibitor alpha in regulating plasticity of hippocampal neurotransmission.

Rab GDP dissociation inhibitor alpha (Rab GDIalpha) is a regulator of the Rab small G proteins implicated in neurotransmission, and mutations of Rab GDIalpha cause human X-linked mental retardation associated with epileptic seizures. In Rab GDIalpha-deficient mice, synaptic potentials in the CA1 region of the hippocampus displayed larger enhancement during repetitive stimulation, which was apparently opposite to the phenotype of Rab3A-deficient mice. Furthermore, the Rab GDIalpha-deficient mice showed hypersensitivity to bicuculline, an inducer of epileptic seizures. These results suggest that Rab GDIalpha plays a specialized role in Rab3A recycling to suppress hyperexcitability via modulation of presynaptic forms of plasticity.

Silicone granuloma on the entry points of acupuncture, venepuncture and surgical needles.

We describe a case of epithelioid granuloma on the entry points of needles used for acupuncture, venepuncture and for taking skin biopsy. The acupuncture needles used at each session were silicone coated. Silicon was detected in the vacuoles of macrophages and multiple nucleated giant cells by X-ray microanalysis. To our knowledge, this is the first case of silicone granuloma arising on the entry points of acupuncture, venepuncture and surgical needles.

Evidence for the presence of 1,2-cyclic acetal type sn-glycero-3-phosphoethanolamines in the sea anemone, Actiniogeton sp.

Five 1,2-cyclic acetal-type sn-glycero-3-phosphoethanolamines (CGPE) were isolated in a pure state from the sea anemone, Actiniogeton sp. (Coelenterata). Their structures, including the absolute configurations, have been determined on the basis of chemical and spectral data to be so-called Feulgen's acetalphosphatides, which have been regarded as artifacts derived from original plasmalogens. We examined whether these CGPE are intact constituents in the animal tissues and obtained reliable confirmation that CGPE are normally present in the sea anemone.

Laser-assisted myringoplasty--technical aspects.

At lower power, lasers fuse collagen fibres and weld tissues. Welded collagen fibres make a solid bond and allow tissue growth along the bonded edges. Our aim was to study applicability of lasers in myringoplasty. We used a KTP-532 laser in outpatient myringoplasty. The laser beam was delivered through a micromanipulator connected to a microscope or through a 200-400-micron silica fibre. The perichondrium was used for transplantant and harvested from the tragus. The margins of the perforation in the eardrum were evaporated, with the laser operating in a continuous mode at 2-4 W. The middle ear was filled with gelfilm to provide support for the transplant. The perichondrium was placed under the margins of the tympanic membrane and lazed at low power (0.2-1.5 W) in continuous mode. In pale tissues, venous blood, methylene blue or fluorescein was used to enhance the tissue admittance of laser energy. Surgical failures were linked to thermal tissue damage due to excessive energy during lazing. In two cases, visibility via microscope into the anterior edge was not complete and the transplantant did not adhere in the relatively limited area. One patient had epidermal growth under the tympanic membrane and developed local cholesteatoma. Laser-assisted myringoplasty provides several advantages over traditional myringoplasty: it is minimally invasive, no manipulation of the ossicles is needed and it is convenient in anterior perforations, where it can be done endoscopically. We prefer a fibre delivery system to a micromanipulator, as lazing with endoscopes is possible and thermal damage is easier to prevent.