Purification and characterization of sea squirt alpha-N-acetylgalactosaminidase

Sea squirt alpha-N-acetylgalactosaminidase was purified to homogeneity. Its molecular weight was estimated to be approximately 160,000 by gel filtration and 40,000 by SDS-polyacrylamide gel electrophoresis (SDS-PAGE) under reducing condition. The chromatographic and electrophoretic behaviors indicated that the enzyme was composed of four subunits. The optimum pH of the enzyme reaction was about 4.0 at 37 degrees C, while the enzyme was stable in the range of pH 5.0 to 6.0 during 4 h preincubation at 37 degrees C. Although the enzyme (0.1 unit) was stable at 0 degrees C for 30 min in the presence of 7.5 mM metal ions (Al3+, Ba2+, Ca2+, K+, Mn2+, Pb2+, Sr2+, and Zn2+), almost 40% of the enzyme activity was lost in the presence of Cu2+, Hg2+, monoiodoacetic acid, and EDTA. The enzyme hydrolyzed aryl N-acetyl-alpha-D-galactosaminide as well as GalNAcalpha1(-->4GalNAcalpha1-->)n 4GalNAc-p-aminobenzoic acid ethyl ester (ABEE) (n = 1-4), but GalNAcalpha1-->4GalNAc-ABEE only scarcely. Furthermore, an allergenic pentasaccharitol ABEE derivative, GalNAcalpha1-->2Fucalpha1-->3(GalNAcbeta1-->4) GlcNAcbeta1-->2(3-acetoamido-3-deoxy)L-threose-ABEE, the minimum structural unit for the sea squirt allergenicity was hydrolyzed to 95 mol% for 72 h incubation with the enzyme. The enzyme could be utilized as a powerful tool for the structural analyses of the carbohydrate epitopes of the sea squirt allergen molecules.     

Enhancement of the thermoelectric figure of merit in p- and n-type Cu/Bi-Te/Cu composites

The resultant thermoelectric properties of welded Cu/Bi-Te/Cu composites were measured at 298 K as a function of relative thickness x of Bi-Te compound by changing the interval s between two thermocouples and compared with those calculated as a function of x by treating it as an electrical and thermal circuit. These composites were prepared by welding with eutectic solder of Pb-Sn, after one end surface of the as-grown p- and n-type Bi-Te ingots were plated with Ni. It was found that the observed ZT of composites has a local maximum at an optimum x even when s was changed, as in the case of Cu/Bi-Sb/Cu and Ni/Bi-Sb/Ni composites with various relative thicknesses. Appearance of a local maximum in ZT is owing to the barrier thermo-emf generated by a sharp temperature drop at the interface between Bi-Te compound and copper. It may be caused by the separation of non-equilibrium carriers at the interface between them. The observed maximum ZT values at 298 K of the p- and n-type composites reached surprisingly great values of 1.53 and 1.66 at x=0.98, which correspond to about twice as large as those of commercially utilized Bi-Te compounds. This enhancement of ZT is available for generators, but may be not utilizable as a Peltier module. The composite materials were thus found to be utilizable as useful means of further increase in ZT of macroscopic bulk materials.     

Reliable indirect competitive ELISA used for a survey of ochratoxin A in green coffee from the North of Paraná State, Brazil.

The performance of an indirect competitive enzyme-linked immunosorbent assay (ic-ELISA) based on a monoclonal antibody (mAb) for ochratoxin A (OTA) detection was evaluated in a comparative study with high-performance liquid chromatography (HPLC) analysis using 68 freshly harvested coffee samples from the North of Paraná State, Brazil. The anti-OTA mAb showed high specificity and low cross-reactivity with OTA analogues (OTB and OTalpha), but cross-reacted with OTC. This ic-ELISA showed a detection limit of 3.75 ngg-1 sample, when compared to 0.80 ngg-1 by HPLC, with an ic-ELISA/HPLC correlation coefficient of 0.90. As regards OTA analysis of these coffee samples, natural contamination was detected in 10 samples (14.7%) by both methods, where the ic-ELISA values (range 3.9-7.3 ngg-1) were 1.1 to 1.6-fold higher than HPLC data (2.7-4.7 ngg-1). Five samples (7.4%) were OTA positive (range 0.84-1.30 ngg-1) only by HPLC assay, probably due to the higher detection limit reached by ic-ELISA. OTA was undetectable in 53 samples (77.9%) by both methods, while all positive samples (range 0.84-7.30 ngg-1) showed OTA levels lower than 8 ngg-1 (maximum limit recommended by the European Union). The matrix interference of green coffee was minimized by dilution of sample extracts before carrying out the ELISA assay. This mAb-based ic-ELISA can be effectively applied for OTA screening in coffee, because it is simple, sensitive and sample preparation is easy.     

Hearing recovery in sudden deafness with profound hearing loss

To more completely describe acromion morphology and its relationship to impingement syndrome, we performed three-dimensional magnetic resonance imaging (N = 111) or computed tomography (N = 27) on 132 symptomatic shoulders. The mean patient age was 46.2 years (range, 14 to 86). Four parameters were evaluated: the angle of anterior slope of the acromion in the midsagittal and lateral-sagittal planes, lateral acromial angulation in the coronal plane, and the presence or absence of medial encroachment in the acromioclavicular joint. Twenty-five asymptomatic age-matched shoulders were used as controls. All imaging data were combined because no significant differences existed between the two imaging techniques. The mean acromion angle was 19.4 degrees in the midsagittal plane and 20 degrees in the lateral-sagittal plane. In the coronal plane, 97 (73%) acromions were neutral and 35 (27%) were downward sloping. Medial encroachment was present in 31 (24%) shoulders. Age distribution from the 2nd to 8th decade demonstrated a consistent and gradual transition from a flat acromion in the younger decades to a more hooked acromion in the older decades that was significant in both the midsagittal and lateral-sagittal planes. Furthermore, a greater percentage of patients were found to have downward angulating acromions with increasing age. Ninety-eight patients (74%) had stage II or III impingement. Of these shoulders, 39 (40%) had type I acromions, 51 (52%) type II, and 8 (8%) type III. Twenty-eight of 33 acromions with coronal lateral downward sloping had impingement, and all 31 shoulders with medial encroachment had impingement.     

Nucleotide sequencing analysis of the 146-kilobase segment around the IkBL and MICA genes at the centromeric end of the HLA class I region

The biomechanics of medialization laryngoplasty are not well understood. An excised canine larynx model was used to test the effects of various sized silicon implants. The vocal fold length, position, and tension were measured. Medialization laryngoplasty did not affect vocal fold length. At the mid-membranous vocal fold, larger shims resulted in greater medialization and tension. Medialization laryngoplasty neither medialized nor stiffened the vocal process to resist lateralizing forces. We conclude that medialization laryngoplasty provides bulk and support for defects of the membranous region of the vocal fold, but does not appear to close a posterior glottal gap. The selection of a surgical procedure to treat glottal incompetence should take into account the unique biomechanical properties of the anterior (membranous vocal folds) and posterior (cartilaginous portion) glottis.     

Clinical significance of natural killing activity in patients with advanced lymphoma

OBJECTIVE: (1) To determine which facial landmarks show the greatest movement during specific facial animations and (2) to determine the sensitivity of our instrument in using these landmarks to detect putatively abnormal facial movements. DESIGN: Movements of an array of skin-based landmarks on five healthy human subjects (2 men and 3 women; mean age, 27.6 years; range, 26 to 29 years) were observed during the execution of specific facial animations. To investigate the instrument sensitivity, we analyzed facial movements during maximal smile animations in six patients with different types of functional problems. In parallel, a panel was asked to view video recordings of the patients and to rate the degree of motor impairment. Comparisons were made between the panel scores and those of the measurement instrument. RESULTS: Specific regions of the face display movement that is representative of specific animations. During the smile animation, landmarks on the mid- and lower facial regions demonstrated the greatest movement. A similar pattern of movement was seen during the cheek puff animation, except that the infraorbital and chin regions demonstrated minimal movement. For the grimace and eye closure animations, the upper, mid-facial, and upper-lip regions exhibited the greatest movement. During eye opening, the upper and mid-facial regions, excluding the upper lip and cheek, moved the most, and during lip purse, markers on the mid- and lower face demonstrated the most movement. We used the smile-sensitive landmarks to evaluate individuals with functional impairment and found good agreement between instrument rankings based on the data from these landmarks and the panel rankings. CONCLUSION: The present method of three-dimensional tracking has the potential to detect and characterize a range of clinically significant functional deficits.     

Osteoclastogenesis inhibitory factor exhibits hypocalcemic effects in normal mice and in hypercalcemic nude mice carrying tumors associated with humoral hypercalcemia of malignancy

In the present study, we attempted to determine the extent to which an anterior capsulorrhexis carried out during cataract surgery contracts postoperatively. The size of the continuous curvilinear capsulorrhexis (CCC) was measured at the end of surgery and at the final follow-up examination in 52 eyes of 40 patients who underwent phacoemulsification/aspiration and implantation of an intraocular lens (IOL) with 6-mm optics. Images obtained by video during surgery and by slit-lamp microscopy were used to determine the CCC size with reference to the optics of the IOL. The average CCC size was 5.0 mm at the end of surgery; it contracted to 4.4 mm by the final postoperative examination. The contraction of the anterior capsule progressed rapidly in the first 50 days following surgery and then gradually thereafter. The extent of contraction was greater in older patients. The smaller the CCC size immediately after surgery, the more rapidly it contracted. The size of the capsulorrhexis contracted an average of 22% following cataract surgery. The contraction rate tended to be higher in older patients. The results suggest that the capsulorrhexis carried out in cataract surgery for elderly patients should be sufficiently large, but not larger than the optics size of the IOL.     

Experimental Facsimile Communication System on Packet Switched Data Network

The use of the public data network for facsimile communication has remarkable advantages with regard to high speed and reliable transmission and, in addition, it is well applicable to provide value added facilities for future service extension. The Research and Development Laboratories of Kokusai Denshin Denwa (KDD) Company, Ltd. have developed an experimental facsimile communication system on the packet switched data network (PSDN). The system is composed of packet switching exchanges (PSE), high speed digital facsimile terminal equipment, a facsimile communication processor (FCP), and facsimile packet assembly/ disassembly equipment (FAX-PAD). The facsimile equipment in the system adopts a two-dimensional modified READ code and has the interface protocol to the PSDN. The FCP provides the store-and-forward facilities to provide such services as delayed delivery, multiaddress delivery, and mailbox service. The FAX-PAD realizes the interworking between facsimile equipments on the public switched telephone network (PSTN) and those on PSDN. After various experiments including international field tests, it was confirmed that the system is useful for future advanced facsimile service.     

Ribotyping and a study of transmission of Staphylococcus aureus collected from food preparation facilities

Food poisoning from Staphylococcus aureus is sometimes caused by improper handling of food items in food preparation facilities. Prevention of contamination by employees is particularly important in facilities where a significant amount of food preparation is performed by hand. Some experiments have been performed to describe bacterial cross-contamination in the food preparation process, but there have been few studies of cross-contamination in actual food preparation facilities. Aiming to shed light on the transmission of S. aureus in food preparation facilities, this study collected samples of 66 strains of this bacterium from the fingers of food preparation staff, foodstuffs, prepared foods, cooking utensils, and cooking equipment and typed them with the ribotyping method. S. aureus from the same ribogroup was detected on the hands of a study participant, a faucet, knife, frying pan, and a salad, indicating that bacteria found on the hands of the study participant was transmitted to cooking utensils and prepared foods. Transmission (from a faucet to a frying pan handle) of bacteria by another person, a third party, was also detected.     

Mating-induced mating-type cassette conversion in Saccharomyces cerevisiae

We have previously reported that the HMRa-bearing restriction fragment of a rho degrees sir4-11 strain (HMLalpha-MATalpha-HMRa), which acts as an alpha-mater because of being rho degrees , changes its electrophoretic mobility when the strain mates with a certain group of a-mating strains (HMLalpha-MATa-HMRa). In this study, we found that the sir4-11 strain being rho degrees was not essential for this phenomenon and also that the altered form of the fragment contained HMRalpha in place of HMRa. Furthermore, we observed conversion of HMLa to HMLalpha in the cross in which a sir4-11 HMLa-MATalpha-HMRalpha strain was mated with a representative of the above-mentioned a-mating strain. In addition, when this a-mating strain was mated with a SIR(+) HMLalpha-MATa-HMRalpha strain, the resultant diploid was found to be HMLalpha/HMLalpha MATa/MATalphaHMRa/HMRalpha, indicating that conversion of MATa to MATalpha had taken place in the course of mating. From all these observations, we conclude that there is a group of S. cerevisiae strains that carries factor(s) that induces conversion of a mating-type cassette of the mating partner to alpha mating-type cassette and that this mating type cassette conversion takes place in all three mating type loci, HML, MAT and HMR, if the loci are in the non-silenced condition.