Evaluation of PCR-based assay for diagnosis of spotted fever group rickettsiosis in human serum samples

A nested PCR assay was developed for the detection of spotted fever group (SFG) rickettsiae in serum samples. The assay was based on specific primers derived from the rickettsial outer membrane protein B gene (rompB) of Rickettsia conorii. An SFG rickettsia-specific signal is obtained from R. akari, R. japonica, R. sibirica, and R. conorii. Other bacterial species tested did not generate any signal, attesting to the specificity of the assay. As few as seven copies of the rompB gene of R. conorii could be detected in 200 microl of serum sample. The assay was evaluated with a panel of sera obtained from patients with acute-phase febrile disease tested by immunofluorescent antibody assay (IFA). The SFG rickettsia-specific DNA fragment was detected in 71 out of 100 sera, which were proven to have immunoglobulin M antibodies against SFG rickettsial antigen by IFA. The results were further confirmed by restriction fragment length polymorphism and sequencing analysis of the DNA fragments. The results indicated that this PCR assay is suitable for the diagnosis of spotted fever group rickettsiosis in Korea.     

Isolation and characterization of human and rabbit sperm tail fibrous sheath

Using mechanical and chemical dissection methods, fibrous sheath was isolated both from normal ejaculated human spermatozoa and from rabbit cauda epididymal spermatozoa. The same techniques did not produce a pure preparation of fibrous sheath from ejaculated rabbit spermatozoa, suggesting that further cross-linking and stabilization of sperm structures occurs in response to components of the seminal plasma. The isolation procedures were monitored by phase contrast microscopy and the purity of the fibrous sheath was verified by electron microscopy. Sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) of isolated human fibrous sheath revealed at least 14 protein bands of which the most intensely stained were of molecular weight 84, 72, 66.2, 57, 32 and 28.5 kDa. The rabbit fibrous sheath revealed at least 10 protein bands, of which the most intensely stained were 35.2, 32.7 and 28.5 kDa. The amino acid composition of the purified fibrous sheath from human and rabbit spermatozoa was similar, being high in aspartic acid and/or asparagine and glutamic acid and/or glutamine, serine, alanine, leucine, lysine and glycine, but low in histidine, tyrosine and isoleucine. This composition is similar to that reported for the rat and suggests that mammalian sperm tail fibrous sheaths are composed of similar types of proteins, although there are apparent differences in protein components between species.      

Feulgen reaction study of novel threadlike structures (Bonghan ducts) on the surfaces of mammalian organs

Threadlike structures on the surfaces of internal organs, which are thought to be part of the Bonghan duct system, were first reported about 40 years ago, but have been largely ignored since then. Recently, they were rediscovered, and in this study we discuss the Feulgen reaction that specifically stains DNA in order to identify these structures on the surface of rabbit livers as part of the Bonghan system. The distribution, shapes, and sizes of their nuclei are found to be similar to those of intravascular threadlike structures. The endothelial nuclei are rod-shaped, 10-20 mum long, and aligned in a broken-line striped fashion. The threadlike structure consists of a bundle of several subducts, which is a characteristic feature of Bonghan ducts and distinguishes them morphologically from lymphatic vessels. In addition, the Feulgen reaction clearly demonstrates that the subducts pass through a corpuscle, which is usually irregular or oval-shaped and is connected to two or several threadlike structures that form a web on the surfaces of organs. Furthermore, spherical granules of about 1 mum in diameter are detected in the subducts. These granules were well stained by using the Feulgen reaction, which implies that they contain DNA. According to previous reports, a granule is a type of microcell and plays an essential role in the physiology and therapeutic effect of the Bonghan system and acupuncture. This role has yet to be elucidated.     

Colorimetric microwell plate hybridization assay for detection of amplified Mycobacterium tuberculosis DNA from sputum samples.

We developed a colorimetric microwell plate hybridization assay (CoMPHA) for the specific detection of 5'-biotinylated amplified Mycobacterium tuberculosis DNA. The optical densities of the CoMPHA corresponded to the initial amounts of purified template DNA. Here, we show that the CoMPHA is useful in distinguishing the PCR-positive and PCR-negative samples.     

Multifocal Parotid Oncocytic Cystadenomatosis: A Rare Process that Can Mimic Malignancy

Multifocal cystic oncocytosis (MCO) is a rare, benign process accounting for approximately 0.1% of salivary gland lesions. Salivary oncocytosis is characterized by multiple unencapsulated solid nodules of oncocytic cells derived from transformed striated ducts. MCO is a variant of salivary oncocytosis which manifests as cystically dilated striated ducts. It is difficult to obtain a definitive preoperative diagnosis of MCO; therefore, these lesions are commonly treated with surgery. We report the unique case of a 66-year-old male who previously underwent a superficial left parotidectomy for a pleomorphic adenoma. Four years later, he presented with clinical and radiographic suspicion of a multifocal recurrent pleomorphic adenoma. The patient subsequently underwent a revision parotidectomy. However, final pathology confirmed a diagnosis of MCO. Although MCO is commonly treated with surgery due to lack of a definitive preoperative diagnosis, surgery is unnecessary outside of diagnostic, functional or cosmetic considerations. Thus, if a patient with parotid oncocytosis treated by superficial parotidectomy develops disease re-manifestation in the residual deep lobe, further surgery is not indicated. There is no risk of malignant progression in this process. We report on this unusual entity as it may mimic salivary malignancy or, as in this case, recurrence of benign disease.     

Structure activity relationship of ar-turmerone analogues

For the analysis of structure activity relationship of ar-turmerone analogues, the compounds containing the various substituents on the phenyl ring and 1(or 2)-naphthyl group in the place of phenyl of ar-turmerone were prepared and tested their cytotoxicity against HL-60, K-562, and L1210 leukemia cellsin vitro. The substituents at para position are methoxy, phenoxy, methyl, trifluoromethyl, fluoro, and chloro. Atmeta position methoxy, methyl, trifluoromethyl, or chloro groups and atortho position methoxy or chloro group were introduced. Against HL-60 and K-562 cells, ED₅₀ values of the analogues are ranged from 0.8 to 30.0 μg/ml. Against L1210 cell, these are located more than 20.0 μg/ml. However, 5-carboethoxy-2-methyl-6-(1-naphthyl)-2-octen-4-one (5n) possesses ED50 valuses 0.8, 2.1, 6.5 μg/ml against HL-60, K-562, L1210 cells, respectively. The electronic nature of the subsituents on phenyl ring of ar-turmerone dose not affect the biological activity. Therefore the flat structure of aromatic portion of ar-turmerone analogues is the more important factor for their activity rather than its electronic nature. The potentiation of the cytotoxicity with the enlargement of aromatic ring region also supports the importance of the plane structure of this area. The restriction of the single bond rotation between C-6 and aromatic ring through the introduction of substituents at theortho position of phenyl ring and the increment of size of alkyl group at C-6 position enhances the activity. Therefore the effective conformation should be the one having the orthogonal arrangement between the aromatic ring and the side chain.     

c-Fos basal immunoreactivity decreases in rat spinal cord during normal ageing

The pattern of distribution in rat spinal cord and changing pattern during normal ageing of c-Fos expression were investigated by immunohistochemical staining in male Sprague-Dawley rats at the age of 1 week, 5 months and 2 years. c-Fos immunoreactivity was observed diffusely in gray matters in neonatal rats, preferentially located in deep dorsal horn and around central canal. Compared with those of neonatal rats, these cells decreased prominently in adult rats. In aged rats, immunoreactive cells were not seen in any segments. c-Fos immunoreactivity in spinal cord may be related to stress response, functional differentiation, and in part, neuronal death with target dependence. In conclusion, we demonstrated for the first time that c-Fos expression patterns change during normal ageing.     

Replantation of avulsed scalps and secondary aesthetic correction

Five patients with avulsed scalps were treated with replantation between 1992 and 1998. All patients were women age 20 to 36 years. The percentage of the avulsed scalp ranged from 50% to 100% of the whole scalp. The vessels chosen for anastomosis were the superficial temporal artery, occipital artery, and superficial temporal vein. A vein graft harvested from the cephalic vein of the forearm was performed on the venous and arterial sides in 1 patient. Two patients experienced complete survival of the replanted scalp. Three patients showed 40%, 50%, and 80% survival areas, with the remaining defects resurfaced as split-thickness skin grafts. Six months later, the scar areas in the last 3 patients were reconstructed with an expansion of the normal or replanted scalp. The follow-up period ranged from 1 to 7 years. In 4 patients a partial return of sensation in the replanted scalp and motor function of the frontalis muscle were observed. All patients were satisfied with the aesthetic results of their surgery.