Cell-substrate interactions in cnidaria

Studies on morphogenesis and regeneration in cnidarians have a long history, and the importance of cell-ECM (extracellular matrix) interactions for these processes has been well recognized and studied since the middle of the 20th century. Cnidarians have a life cycle with a larva, a polyp, and often a medusa generation. In the medusa, the ECM (mesoglea) is very prominent and essentially shapes the animal. In the larva and the polyp, the ECM is a thin layer. Some of the ECM components known from vertebrates have been identified in cnidarians by immunohistochemistry, electron microscopy, rotary shadowing, biochemistry, and molecular cloning. In vivo and in vitro experiments suggest that the cnidarian ECM plays a role in cell migration and morphogenesis comparable to that known from other developmental systems. In the fresh water polyp Hydra, regeneration of body patterns and migration of nematocytes seems to require the presence of ECM ligands and the corresponding cell receptors. In hydrozoan medusae, DNA replication and the stability of the differentiated state of isolated tissue can be influenced by altering the properties of the ECM substrate. When cultured, most cnidarian cells survive only when attached to ECM substrates, they rarely divide and die within short times.     

A practical method of measuring scattering characteristics of apyramidal absorber

A free-space, standing-wave technique at oblique incidence is studied as a method of measuring the scattering characteristics of a pyramidal or wedge-shaped absorber. Using this method, the standing-wave ratio in front of the sample from waves with electric fields of equal amplitude are scattered in three different directions, and their scattering angles are easily measured over a wide range of frequencies and angles of incidence. No special measuring apparatus or complicated procedures are required with this method. It is found that the standing-wave ratio around 1.1 (magnitude of the scattering coefficient=0.04762) is less than ±20% in terms of scattering coefficient for incident angles ranging from 0° to 80°. The error in the scattering angle is less than 4° for the 0° scattered wave and less than ±3° for all other scattering angles     

Molecular cloning and chromosomal mapping of mouse intronless myc gene acting as a potent apoptosis inducer

CD38, a transmembrane glycoprotein widely expressed in vertebrate cells, is a bifunctional ectoenzyme catalyzing the synthesis and hydrolysis of cyclic ADP-ribose (cADPR). cADPR is a universal second messenger that releases calcium from intracellular stores. Since cADPR is generated by CD38 at the outer surface of many cells, where it acts intracellularly, increasing attention is paid to addressing this topological paradox. Recently, we demonstrated that CD38 is a catalytically active, unidirectional transmembrane transporter of cADPR, which then reaches its receptor-operated intracellular calcium stores. Moreover, CD38 was reported to undergo a selective and extensive internalization through non clathrin-coated endocytotic vesicles upon incubating CD38(+) cells with either NAD+ or thiol compounds: these endocytotic vesicles can convert cytosolic NAD into cADPR despite an asymmetric unfavorable orientation that makes the active site of CD38 intravesicular. Here we demonstrate that the cADPR-generating activity of the endocytotic vesicles results in remarkable and sustained increases of intracellular free calcium concentration in different cells exposed to either NAD+, or GSH, or N-acetylcysteine. This effect of CD38-internalizing ligands on intracellular calcium levels was found to involve a two-step mechanism: 1) influx of cytosolic NAD+ into the endocytotic vesicles, mediated by a hitherto unrecognized dinucleotide transport system that is saturable, bidirectional, inhibitable by 8-N3-NAD+, and characterized by poor dinucleotide specificity, low affinity, and high efficiency; 2) intravesicular CD38-catalyzed conversion of NAD+ to cADPR, followed by outpumping of the cyclic nucleotide into the cytosol and subsequent release of calcium from thapsigargin-sensitive stores. This unknown intracellular trafficking of NAD+ and cADPR based on two distinctive and specific transmembrane carriers for either nucleotide can affect the intracellular calcium homeostasis in CD38(+) cells.     

Sunlight mutagenesis: changes in mutational specificity during the irradiation of phage M13mp2

BACKGROUND: The accumulation of fibrillar deposits of amyloid beta-peptide (Abeta) in brain parenchyma and cerebromeningeal blood vessels is a key step in the pathogenesis of Alzheimer's disease. In this report, polymerization of Abeta was studied using fluorescence correlation spectroscopy (FCS), a technique capable of detecting small molecules and large aggregates simultaneously in solution. RESULTS: The polymerization of Abeta dissolved in Tris-buffered saline, pH 7.4, occurred above a critical concentration of 50 microM and proceeded from monomers/dimers into two discrete populations of large aggregates, without any detectable amount of oligomers. The aggregation showed very high cooperativity and reached a maximum after 40 min, followed by an increase in the amount of monomers/dimers and a decrease in the size of the large aggregates. Electron micrographs of samples prepared at the time for maximum aggregation showed a mixture of an amorphous network and short diffuse fibrils, whereas only mature amyloid fibrils were detected after one day of incubation. The aggregation was reduced when Abeta was incubated in the presence of Abeta ligands, oligopeptides previously shown to inhibit fibril formation, and aggregates were partly dissociated after the addition of the ligands. CONCLUSIONS: The polymerization of Abeta is a highly cooperative process in which the formation of very large aggregates precedes the formation of fibrils. The entire process can be inhibited and, at least in early stages, partly reversed by Abeta ligands.     

Role of the sulphate assimilation pathway in utilization of glutathione as a sulphur source by Saccharomyces cerevisiae.

Mutants unable to grow on medium containing glutathione as a sole source of sulphur (GSH medium) were isolated from Saccharomyces cerevisiae strains carrying met17(deficiency of O-acetylserine and O-acetylhomoserine sulphydrylase). They were defective in the high-affinity glutathione transport system, GSH-P1. Newly acquired mutations belonged to the same complementation group, gsh11. However, it became apparent that gsh11 conferred the mutant phenotype not by itself but in collaboration with met17. Moreover, mutations conferring the defect in sulphate assimilation made the cell unable to grow on GSH medium in collaboration with gsh11. From this finding, we propose that the sulphate assimilation pathway acts as a sulphur-recycling system and that this function is especially vital to the cell when the supply of glutathione is limited.     

Primary adenocarcinoma of the renal pelvis with special reference to histochemical observations

A case of adenocarcinoma of the renal pelvis is presented. The patient was an 84-year-old man suffering from long-standing right-sided nephrolithiasis. Surgical resection of the right kidney revealed adenocarcinoma with slight stromal invasion. A tubulovillous adenoma, which was morphologically similar to an adenoma of the large intestine, was also found adjacent to the adenocarcinoma. The pelvic epithelium neighboring the lesion revealed intestinal metaplasia. Histochemical studies revealed that the tumor in the patient and adenocarcinomas or adenomas of the large intestine have similar properties of cytoplasmic mucin. These findings suggest that the epithelium with intestinal metaplasia may have developed into the adenoma and finally transformed into the adenocarcinoma. In addition, only tumor cells with severe atypia, most of which morphologically corresponded to adenocarcinoma, demonstrated positive nuclear staining for anti-p53. This suggests that p53 may play an important role in the malignant transformation of adenomas into adenocarcinomas, as is the case in the large intestine.     

Self-assessed quality of mastication in relation to remaining teeth and self-assessed health status in elderly persons

To explore factors related to quality of mastication, information about teeth status and self-assessed health status were obtained by a self-administered questionnaire from participants in a dental screening program. Invitation letters for participation in the program were distributed by members of the neighborhood association to 1.275 residents of a town in Gifu prefecture aged 65 years or older. Participation rate in the dental screening program was 40.5%. A total of 513 participants (245 males and 268 females) completed the program and the questionnaire. The results were as follows. 1. Overall, 32% had 20 teeth or more and 88.4% had bridge(s) and/or denture(s). 2. The mean number of remaining teeth were 12.9 for males and 12.5 for females. There was no statistical difference in the number of remaining teeth between males and females. The mean numbers of remaining teeth among those aged 65-69 years was 15.7 and two thirds of them had lost the Lower First Molar. 3. Those who reported to be satisfied with their present mastication were 83.3% of men and 86.3% of women. The proportions were not influenced by age or sex. 4. Quality of mastication was related to self-reported good physical condition' and not easily tired'. 5. The proportion of those satisfied with their mastication was large in the groups with remaining teeth index 0-8 and > or = 17, and small in the group with remaining teeth index 9-16, showing a U-shaped curve for satisfaction rate against remaining teeth index. These results suggest a relationship of quality of mastication on health status. The importance of primary dental care for the elderly persons is underlined.     

Hyaluronic acid is increased in the skin and urine in patients with amyotrophic lateral sclerosis

We performed morphological studies of skin and measured glycosaminoglycans in the urine from patients with sporadic amyotrophic lateral sclerosis (ALS) and control subjects. The wide spaces separating collagen bundles reacted strongly with alcian blue stain in ALS patients and stained more markedly as ALS progressed. Staining with alcian blue was virtually eliminated by Streptomyces hyaluronidase. The urinary excretion of hyaluronic acid (HA) (mg/day) was significantly increased (P < 0.01) in ALS patients compared with that of control subjects, and there was a significant positive correlation between the excreted amount of HA and the duration of illness in advanced ALS patients with a duration of more than 2 years from clinical onset (r = 0.72, P < 0.02). We suggest that sporadic ALS includes a metabolic disorder of HA in which an accumulation of HA in the skin is linked to an increased urinary excretion of HA.     

A randomized crossover study of an oral appliance vs nasal-continuous positive airway pressure in the treatment of mild-moderate obstructive sleep apnea

STUDY OBJECTIVE: To compare efficacy, side effects, patient compliance, and preference between oral appliance (OA) therapy and nasal-continuous positive airway pressure (N-CPAP) therapy. DESIGN: Randomized, prospective, crossover study. SETTING: University hospital and tertiary sleep referral center. PATIENTS: Twenty-seven unselected patients with mild-moderate obstructive sleep apnea (OSA). INTERVENTIONS: There was a 2-week wash-in and a 2-week wash-out period, and 2 x 4-month treatment periods (OA and N-CPAP). Efficacy, side effects, compliance, and preference were evaluated by a questionnaire and home sleep monitoring. MEASUREMENTS AND RESULTS: Two patients dropped out early in the study and treatment results are presented on the remaining 25 patients. The apnea/hypopnea index was lower with N-CPAP (3.5 +/- 1.6) (mean +/- SD) than with the OA (9.7 +/- 7.3) (p < 0.05). Twelve of the 25 patients who used the OA (48%) were treatment successes (reduction of apnea/hypopnea to <10/h and relief of symptoms), 6 (24%) were compliance failures (unable or unwilling to use the treatment), and 7 (28%) were treatment failures (failure to reduce apnea/hypopnea index to <10/h and/or failure to relieve symptoms). Four people refused to use N-CPAP after using the OA. Thirteen of the 21 patients who used N-CPAP were overall treatment successes (62%), 8 were compliance failures (38%), and there were no treatment failures. Side effects were more common and the patients were less satisfied with N-CPAP (p < 0.005). Seven patients were treatment successes with both treatments, six of these patients preferred OA, and one preferred N-CPAP as a long-term treatment. CONCLUSIONS: We conclude that OA is an effective treatment in some patients with mild-moderate OSA and is associated with fewer side effects and greater patient satisfaction than N-CPAP.