Distribution of indigenous bacteria in the digestive tract of conventional and gnotobiotic rats.

The localization and population levels of the indigenous bacterial flora of conventional rats were investigated by cultural and histological techniques. Lactobacilli predominate in the stomach and upper part of the small intestine and associate with keratinized cells of the nonglandular portion of stomach. Mixtures of varying complexity of pure cultures of indigenous bacteria were inoculated into germfree rats. The distribution of these bacteria was examined to investigate the effect of lactobacilli in controlling the composition of other bacterial species in each portion of the digestive tract. In the stomach and the upper part of the small intestine, lactobacilli controlled the population levels of other bacterial species. In the lower part of the small intestine, not only lactobacilli but also the anaerobes which colonized the large bowel influenced the population levels of other bacterial types. Staphylococci isolated from a conventional rat colonized specifically the keratinized cells of the nonsecreting epithelium of the stomach when the rats were free from lactobacilli. This colonization was not observed after inoculation of lactobacilli into the rats.     

Mediolateral intercalation in planarians revealed by grafting experiments.

We investigated how planarians organize their left-right axis by using ectopic grafting. Planarians have three body axes: anteroposterior (A-P), dorsoventral (D-V), and left-right (L-R). When a small piece is implanted into an ectopic region along the A-P and D-V axes, intercalary structures are always formed to compensate for positional gaps. There are two hypotheses regarding L-R axis formation in this organism: first, that the left and right sides of the animal may be recognized as different parts, and L-R intercalation can induce midline structures (asymmetry hypothesis); second, that both sides may have symmetrical positional values, and mediolateral (M-L) intercalation creates positional values along the L-R axis (symmetry hypothesis). We performed ectopic grafting experiments in the head region of the planarian, Dugesia japonica, to examine these hypotheses. A left lateral fragment containing a left auricle was implanted into the medial region of the host. Ectopic structures were always formed only on the left side of the graft, where lateral tissues abutted onto the medial tissues. However, no morphologic change was induced on the right side of the graft, where left-sided tissues faced onto right-sided tissues. Molecular marker analyses indicated that ectopic structures formed on the left side of the graft were induced by M-L intercalation, supporting the "symmetry hypothesis." When the midline tissues were implanted into a lateral region, they induced a complete ectopic head, demonstrating that M-L intercalation may be sufficient to establish the L-R axis in planarians.     

Detection and characterization of simian retroviruses homologous to human T-cell leukemia virus type I

Lymphoid cell lines were established from five different species of monkeys which were positive in antibodies cross-reactive with human T-cell leukemia virus type I (HTLV-I) and were shown to contain provirus sequences homologous to HTLV-I. Gene-specific probes of HTLV-I, gag, pol, env, pX, and LTR, hybridized efficiently with monkey DNAs from these cell lines under stringent conditions, indicating that the proviruses are very similar to HTLV-I along with whole viral genomes. However, the preliminary restriction mapping turned out the difference between simian retroviruses and HTLV-I and also among simian retroviruses. These findings suggest a common ancestor of simian and human retroviruses, but exclude the recent interspecies transmission between monkeys and humans.     

High Frequency of Antibiotic-Associated Diarrhea due to Toxin A-Negative, Toxin B-Positive Clostridium difficile in a Hospital in Japan and Risk Factors for Infection

Patients hospitalized in a hospital with a high incidence of antibiotic-associated diarrhea due to toxin A-negative, toxin B-positive (A–/B+) Clostridium difficile were retrospectively investigated to determine the clinical manifestations and risk factors for infection. Of 77 Clostridium difficile isolates obtained from 77 patients during the 1-year investigation period, 30 were A–/B+ and 47 were toxin A-positive, toxin B-positive (A+/B+). By pulsed-field gel electrophoresis analysis, 23 of the 30 A–/B+ strains were outbreak-related, suggesting nosocomial spread of a single type of bacterium, which mainly affected patients in the wards of respiratory medicine, hematology and neurology. Using regression analysis, three factors were found to be associated with infection by A–/B+ isolates: (i) exposure to antineoplastic agents (P=0.01, odds ratio [OR]=5.1), (ii) the use of nasal feeding tubes (P=0.008, OR=5.2), and (iii) assignment to a certain internal medicine ward (P=0.05, OR=3.0). Between patients with Clostridium difficile-associated diarrhea caused by A–/B+ strains and those with A+/B+ strains, no statistically significant difference was found in body temperature, serum concentration of C-reactive protein, leukocyte count in whole blood, frequency of diarrhea, or type of underlying disease. These results indicate that A–/B+ strains of Clostridium difficile can cause intestinal infection in humans and they spread nosocomially in the same manner as A+/B+ strains.     

Control of upright standing posture during low-frequency linear oscillation

We examined the effects of anteroposterior movement of a sled on human upright standing. Each of six healthy men stood on the platform of a sled in the dark. The sinusoidal acceleration was provided, from 0.02 to 0.04 G, followed by 0.06 and 0.08 G, at a stroke length from 6 to 10 m and then to 14 m. Low acceleration (0.02 and 0.04 G) induced body sway, pivoting on the ankle joint. High acceleration (0.06 and 0.08 G) increased body sway, but the head-neck joint remained locked upright. The electromyographic recordings of the lower leg muscles revealed continuous tonic EMG activities of the gastrocnemius and tibialis anterior muscles at acceleration of 0.02 and 0.04 G, while reciprocal activation was observed at 0.06 and 0.08 G. During head movement, the neck muscles were slightly activated tonically at acceleration of 0.02 and 0.04 G, but they were markedly and tonically activated at 0.06 and 0.08 G. We speculate that the sled oscillation caused body sway in proportion to the acceleration, with the ankle joint playing a principal role. Analysis of neck movement also revealed that the head was held in a fixed upright position, indicating that the vestibulocollic reflex might tonically activate the neck muscles.     

Direct sequencing of a HLA-DRB gene by polymerase chain reaction: sequence variation in DRw8 specificity

The nucleotide sequence of a HLA-DRB gene with a predominant subtype of DRw8 specificity in Japanese (DR8.1) was determined with single-stranded DNA enzymatically amplified by polymerase chain reaction (PCR). The sequence differs at a single amino acid from both of the published DRw8/Dw8.1 and DRw8/Dw8.2 sequences: isoleucine67(AUC) instead of phenylalanine67(TTC) in DRw8/Dw8.1 and serine57(AGC) instead of aspartic acid57(GAT) in DRw8/Dw8.2. On the other hand the DR8.1 and DRw8/Dw8.3 have the same amino acid sequence although one silent nucleotide substitution has occurred between the two sequences. These results indicate that Japanese DR8.1 specificity corresponds to DRw8/Dw8.3. Furthermore, an oligonucleotide probe specific for this sequence was synthesized and hybridized with 33 HLA-typed controls. This probe clearly distinguished the particular subtype from other DRw8 subtypes and specificities.     

The intrinsic vasculature of the lumbosacral spinal nerve roots

The observations made in this study strongly support the concept that spinal nerve roots in general and the human lumbosacral spinal nerve roots in particular are structurally, vascularly, and metabolically unique regions of the nervous system. Peculiarities of their intrinsic vasculature and supporting connective tissue may account for suspected "neuroischemic" responses to pathologic mechanical stresses and inflammatory conditions associated with degenerative disease of the lower spine. It is hoped that the newly described features of the radicular vasa nervorum (to avoid confusion with the dissimilar blood supply of peripheral nerves, the term "vasa radiculorum" might be more accurate) may advance the understanding of certain aspects of lower spine symptomatology and provide some basis for much needed future research.     

Successful Single-Lung Transplant for the Dominant Side: A Case Report

Although single-lung transplant on the side with better lung function is challenging in patients with significantly asymmetrical lung function between the right and left sides, it sometimes can be a realistic option because of the recipient's condition and from the viewpoint of organ sharing. We report our experience with a successful case of single-lung transplant on the side with a pulmonary perfusion ratio of 89%. The transplant was performed with the patient under central venoarterial extracorporeal membrane oxygenation through a clamshell incision, and the patient had an acceptable short- and long-term outcome with a remarkable improvement of lung function.     

Innovative Fixation Technique for Avulsion Fractures of the Calcaneal Tuberosity

Avulsion fractures of the calcaneal tuberosity, although relatively uncommon, occur more frequently in patients with osteoporosis and in the elderly. The results of closed manipulation are poor in these fractures, usually requiring open reduction and internal fixation. However, it is difficult to fix the bone fragment rigidly, because the avulsed bone fragment is small and thin, and the bone quality of the calcaneal body in the elderly is poor. Hence, it is necessary to limit prolonged weight-bearing after the operation. We performed an innovative surgical procedure of suture fixation to the anchor screw in four cases, following which earlier postoperative rehabilitation with full weight-bearing walking and range of motion exercises was possible, and bony union was achieved without repeated displacement of the fragment in all patients. We believe this technique would prove useful in surgical management of calcaneal tuberosity avulsion fractures.