An improved synthesis of 6-azathymidine

The trisaccharides panose and isopanose were prepared in good yield from enzymic hydrolyzates of pullulan. Pullulan was hydrolyzed by the purified alpha amylase preparation of Thermoactinomyces vulgaris R-47. The digest was applied to a carbon-Celite column and eluted with a linear gradient of 1-propanol from 0 to 5%. From the trisaccharide fractions eluted, panose was prepared in about 70% yield. Pullulan was also hydrolysed by purified isopullulanase (EC 3.2.1.57 pullulan 4-glucanohydrolase) of Aspergillus niger ATCC-9642, and isopanose was prepared in about 90% yield by using the same technique as that for the preparation of panose.     

The influence of dental occlusion on electromyographic silent periods

Electromyographic silent periods (EMG SP) and occlusal contact intensity were studied in 31 young patients (9-18 years of age) having different types of malocclusion. EMG SP was induced from bilateral temporalis and masseter muscles by chin tapping during isometric contraction. The simple silent period (SSP), depressed activity (DP) and double silent period (2SP or 3SP) were classified according to the character of silence. The occlusal contact intensity was obtained by using a polymer wafer for clenching and was analysed with a photocclusion analyser. It was found that the average amplitude of temporalis and masseter muscles during clenching was about 400 microV. The mean SSP was around 34 msec. The total SP was around 40 msec in temporalis and 44 msec in masseter. All of the subjects demonstrated one or more DP in ten taps while 18 of them had 2SP or 3SP. The difference in contact intensity between right and left had a strong correlation with the EMG SSP+DP, SSP+2SP3SP and total SP. Anterior-posterior difference was not as strongly related to SP parameters. Less significant correlation was found between SSP and occlusal intensity. It is concluded that the influence of occlusion on EMG SP is basically related to the uneven occlusal contact between the two sides rather than the total occlusal contact intensity or the difference in intensity anterior-posteriorly.     

The effects of practice and visual feedback on mandibular border movements

Twenty healthy subjects were studied on the effects of training on mandibular border movements. Maximum left (LL) and right (RL) lateral excursions, maximum protrusive movement (PT), maximum mouth opening (MO), the difference between left and right excursions (R-L), midline deflection (DF) during opening and closing and midline deviation of the jaw (MOD) at maximum opening position of mandibular border tracing with or without practicing and visual feedback were compared among various sessions. No significant difference has been found on the amount of border extension under the influence of training. However, 70 to 85% of the subjects had some improvement after verbal instruction practicing, while only 50 to 65% of the same subjects showed improvement through visual feedback. It is suggested that doing research related to the jaw border movement on healthy subjects does not have to train them to obtain comparable data. On the other hand, since repeated border tracing in healthy subjects did not worsen the results, practicing or visual feedback training might ascertain a repeatable border tracing.     

Improvements of GC and HPLC analyses in solvent (acetone-butanol-ethanol) fermentation byClostridium saccharobutylicum using a mixture of starch and glycerol as carbon source

A study on the feasibility of using improved computer-controlled HPLC and GC systems was carried out to shorten the time needed for measuring levels of the substrates (glucose, maltose, and glycerol) and products (acetone, butanol ethanol, acetic acid, and butyric acid) produced byClostridium saccharobutylicum DSM 13864 during direct fermentation of sago starch to solvent. The use of HPLC system with a single injection to analyse the composition of culture broth (substrates and products) during solvent fermentation was achieved by raising the column temperature to 80°C. Although good separation of the components in the mixture was achieved, a slight overlap was observed in the peaks for butyric acid and acetone. The shape of the peak obtained and the analysis time of 26.66 min were satisfactory at a fixed flow rate of 0.8 mL/min. An improved GC system was developed, that was able to measure the products of solvent fermentation (acetone, butanol, ethanol, acetic acid, and butyric acid) within 19.28 min. Excellent resolution for each peak was achieved by adjusting the oven temperature to 65°C.     

Single-Column Purification of Syncephapepsin—An Aspartic Proteinase fromSyncephalastrum racemosum

Syncephapepsin is a fungal aspartic proteinase fromSyncephalastrum racemosum.By using the property of syncephapepsin after having increased activity at higher temperature, two rapid purification protocols were developed. In the former case, a crude extract was initially diluted fivefold with an activity assay buffer and heated at 50°C overnight. In this situation, syncephapepsin would digest most of the proteins that the crude extract contained. Subsequently, syncephapepsin of the crude extract was precipitated from 50 to 70% of ammonium sulfate and the preparation was then directly applied to the Superdex 200 HR FPLC column. In this manner, syncephapepsin was rapidly purified to apparent homogeneity within 24 h. In this report, an alternative method of purification is also provided. Compared with the procedure mentioned above, the heating step was proceeded after FPLC chromatography through which the same result was obtained. Using cytochromecand RNase A as substrates, the cleavage sites of both substrates were identified by HPLC peptide mapping. The results showed that syncephapepsin had a broad specificity. Residues recognized to be cleaved were primarily those of trypsin and chymotrypsin and Lys was the most susceptible.     

Preparation of large temporomandibular joint specimens for pathological study

Ten fresh temporomandibular joint (TMJ) specimens about 5 X 4.5 X 3.5 cm in size were removed at autopsy by 5 cuts according to appropriate anatomical landmarks. After routine formalin fixation, the whole-TMJ specimens were wrapped with a thin layer of self-curing resin and then cut with a low speed bone saw along the parasagittal plane predetermined by x-ray guidance. Each specimen was serially cut into 4 to 5 parallel slices of 3 mm thickness, which were then decalcified with 14% EDTA and embedded in paraffin. Histological sections of 5 microns were stained with hematoxylin and eosin. The procedures were accomplished within 20 to 24 days after autopsy. With this technique, the anatomical interrelationships among the various joint components could be maintained and the macroscopic and microscopic topography of the TMJ could be studied in the desired reference plane. Therefore, the corresponding changes among the joint components in a diseased TMJ could be thoroughly examined. This technique was also applicable for the study of large specimens containing both hard and soft tissues.