Quantitative detection of mutant alleles of the K-ras gene with minor groove binder-conjugated fluorogenic DNA probes

Tumor-specific point mutations are stable biomarkers compared with tumor-specific mRNA expression, and are therefore useful to detect occult tumor cells. These mutations have never been used for real-time quantitative polymerase chain reaction (RQ-PCR) assays, because the ability of conventional probes to discriminate between wild-type and mutant alleles is poor. Recently, DNA probes with conjugated minor groove binder (MGB) have been developed. Because of their high melting temperature, these probes achieve high performance in detecting single nucleotide mismatches. Using the MGB technology, we developed a new RQ-PCR system for detecting occult tumor cells in patients with colorectal cancer (CRC), targeting K-ras point mutations. Sixteen MGB-conjugated DNA probes were designed for all previously reported K-ras mutations. The performance of these probes was examined with plasmid DNAs into which K-ras point mutations had been inserted, 32 cancer cell lines and 338 lymph nodes obtained from 15 CRC patients. Fifteen of the 16 MGB probes designed were useful for accurate quantitative assessment, and achieved high sensitivity (1/10(4)-10(5) background cells) and high reproducibility (coefficients of variation <10%). Performance in discriminating single nucleotide mismatches was superior for MGB probes compared with non-MGB probes. We detected a micrometastasis (5.85/10(4) cells equivalent) in one (0.9%) of 110 lymph nodes obtained from 6 patients with K-ras mutations. There was no true false-positive result in 209 lymph nodes obtained from 9 patients without K-ras mutations. The MGB RQ-PCR assay targeting K-ras mutations is an accurate quantitative method for detecting occult tumor cells in CRC.     

Developmental changes in the Collis horizontal reaction analyzed using a three-dimensional video motion analysis system. I. Examination of techniques for inducing Collis horizontal reaction

We used a three-dimensional video motion analysis system to evaluate techniques for inducing the Collis horizontal reaction in 46 infants (total 217 times) from 1-9 months old. Additionally, we examined the safety of different techniques for inducing the Collis horizontal reaction in 20 infants (total 66 times), from 1-3 months old, 40 times by holding the shoulder joint and 26 times by not holding the shoulder joint. Regardless of who performed the examination, the average height that induced the reaction increased from 24.3 to 34.6 cm and the average speed of reaction increased from 28.2 to 43.4 m/s as the age of the infants increased. Additionally, when the Collis horizontal reaction was induced in infants 1-3 months old by holding the shoulder joint of infants, the average fall angle of the head was 7.291 degrees and was not significantly larger than when the reaction was induced by not holding the shoulder joint, which resulted is an average fall angle of the head was 19.302 degrees. However, although the average speed to support the body increased, the fall angle of the head did not increase when the shoulder joint was held. The Collis horizontal reaction is usually not performed in early infancy before head control is acquired, because of the burden to neck caused by the fall of the head and the excess expansion of shoulder joint capsule. However these results suggest that when performing the Collis horizontal reaction in early infancy before head control is acquired, the Collis horizontal reaction can be safely performed by holding the shoulder joint, regardless of the skill of the diagnostician. Therefore, the Collis horizontal reaction is an excellent method to assess infant development, and can be performed at the usual infant health examinations.     

Developmental change of Moro reflex studied with a three-dimensional motion analysis system

We evaluated developmental change of the reaction time of the Moro reflex from birth to three months of age using a three-dimensional motion analysis system (ToMoCo-VM:Tousou system). The reaction time was the shortest in one-month-old babies and became successively prolonged in two and three-month-old babies, showing a U-shaped pattern. The three phase of the Moro reflex, as defined by McGraw, changed with advancing age. The reaction time changed monthly. At the same monthly age, the reaction time was the same irrespective of the phase.     

Spacer effect in the template polycondensation of nucleotide analogs with diamines

The template effect of polymers containing the adenine residue, 6 , bound to a polystyrene backbone via a spacer group, on the polycondensation of active esters of thymine, 1 , and theophylline, 2 , with piperazine was studied in pyridine/methylene chloride or DMF. Template polymers accelerated the polycondensation of 1 with diamines by a factor of 5–20, but the polycondensation of 2 with piperazine showed no considerable template effect.     

Yeast ribosomal proteins: XI. Molecular analysis of two genes encoding YL41, an extremely small and basic ribosomal protein,from Saccharomyces cerevisiae

Summary Two genes encoding ribosomal protein YL41 were cloned from Saccharomyces cerevisiae chromosomal DNA. Both genes contain an uniterrupted region of only 75 nucleotides coding for a protein of 3.3 kD. Within the coding regions the nucleotide sequences are virtually identical, whereas in both the 5-and 3-flanking regions the two genes differ significantly from each other. The deduced protein shows an arginine and lysine content of 68 percent, i.e., 17 out of 25 residues, and the basic residues are evenly distributed over the molecule. When compared to the ribosomal protein sequences currently available no counterpart to YL41 could be found in prokaryotes and it seems likely that YL41 is a eukaryotespecific ribosomal protein.     

Mucosal carcinoma within a colonic diverticulum

We report a case of a mucosal carcinoma and adenoma within a diverticulum in the cecum. Radiographic, endoscopic, and pathologic evaluation of the tumor is presented. Surgical resection was undertaken because of the size and shape of the lesion, risk of perforation, and the possibility of malignancy. A recent review of the literature with respect to clinical signs, diagnosis, growth of the carcinoma, and treatment of tumors around or within diverticula is also presented. A carcinoma or adenoma arising within the diverticulum is very rare. Endoscopic resection of the tumor could entail the risk of perforation, because of the lack of muscular coats in the diverticula. Surgical treatment may be the procedure of choice for lesions near or within the diverticula. Received: November 19, 1998 / Accepted: March 26, 1999     

Do panty liners promote vulvovaginal candidiasis or urinary tract infections?: A review of the scientific evidence

Panty liners are used to absorb light menstrual flow, vaginal discharge, or urine leakage, or to maintain a clean, dry feeling. Allegations that panty liners may trap heat and moisture to promote vulvovaginal candidiasis (VVC) or promote colonization by microbes that contribute to urinary tract infections appear to be unfounded. As reviewed herein, measurements of the impact of panty liners on skin temperature and skin surface moisture had no clinically meaningful effect on cell densities of genital microflora. Epidemiological investigations of a potential link to VVC were either negative or were inconclusive because of confounding factors. Although enteric microbes reside on the vulva and perineum, no evidence exists that panty liner use promotes urethral colonization by enteric microbes. Moreover, a series of 13 randomized prospective trials of panty liners or ultra-thin pads demonstrated no clinically significant adverse effects either on the skin or on isolation frequencies or cell densities of representative genital microflora. Post-market surveillance systems suggest a low incidence of complaints. Evidence from vulvar clinic patients reveals no significant contribution of these products to persistent vulvar symptoms. Taken together, the scientific evidence supports the conclusion that panty liners are safe when used as intended and do not promote VVC or urinary tract infections.