Differences in Gait Patterns of Unilateral Transtibial Amputees With Two Types of Energy Storing Prosthetic Feet

OBJECTIVE: To evaluate if there is a difference in gait pattern when applying two different shapes of energy storing prosthetic feet for trainstibial amputation we conducted a comparative study. Energy storing prosthetic feet for transtibial amputation are increasing in use, but there are few studies that evaluate the effects of the shape of energy storing feet on gait patterns. METHODS: Ten unilateral transtibial amputees were recruited. Two different shapes of dynamic response feet were applied to each subject either 1C30 Trias or 1C60 Triton. The main differences between the two are a split forefoot and the presence of a heel wedge. Spatiotemporal, kinematic, and kinetic data was obtained through gait analysis. Differences between intact and prosthetic side and differences between the two prosthetics were assessed. RESULTS: On a side to side comparison, cadence asymmetry with 1C30 Trias was observed. Ankle plantarflexion at the end of stance and ankle supination at the onset of preswing was smaller with both prosthetic feet compared to the intact side. Other spatiotemporal, kinematic, and kinetic data showed no significant differences in a side to side comparison. In a comparison between the two prosthetics, stance and swing ratio and ankle dorsiflexion through mid-stance was closer to normal with 1C60 Triton than 1C30 Trias. Other spatiotemporal, kinematic, and kinetic data showed no statistically significant differences between prosthetics. CONCLUSION: Both energy storing feet implants showed symmetric gait in unilateral transtibial amputees who are functionally independent in daily living. And 1C60 Triton showed closer to normal gait patterns than 1C30 Trias in our study.

Analysis of Foveal Microvascular Structures Using Optical Coherence Tomography Angiography in Age-stratified Healthy Koreans

PURPOSE: To evaluate foveal avascular zone (FAZ) microvascular structural changes in healthy Korean subjects stratified by age using optical coherence tomography angiography (OCTA). METHODS: Eighty eyes of 43 healthy volunteer subjects who had 20/20 or better best corrected visual acuity without other ocular or systemic disease except vitreous floaters and dry eye syndrome were enrolled and stratified by age group. To measure FAZ size and vascular density (VD) of the superficial capillary plexus (SCP) and deep capillary plexus (DCP), OCTA (DRI OCT Triton, Swept Source OCT, Topcon, Tokyo, Japan) scans were performed over fovea-centered 3 × 3 mm² regions, and then compared with central macular thickness (CMT) and subfoveal choroidal thickness. RESULTS: Mean age of the participants was 46.4 ± 16.1 (20–78). The SCP and DCP FAZ sizes were 0.32 ± 0.11 mm² and 0.41 ± 0.14 mm², respectively. There was a significant difference by age group (p < 0.001, p < 0.001), respectively. The FAZ VD for SCP and DCP was 28.96 ± 3.05% and 33.15 ± 3.64%, respectively. There was no difference between age groups (p = 0.118, p = 0.637). Univariate and multiple linear regression analysis showed that the FAZ size of SCP and DCP was significantly correlated with age (p = 0.039, p = 0.002) and CMT (p = 0.007, p = 0.013), respectively. The SCP and DCP FAZ size were positively correlated with age (R² = 0.279, p < 0.001, R² = 0.344, p < 0.001), and negatively correlated with CMT (R2 = 0.354, p < 0.001, R2 = 0.285, p < 0.001), respectively. CONCLUSIONS: The FAZ size of SCP and DCP increased with age and were negatively correlated with CMT. These results suggest that consideration of age and CMT is important when performing the clinical evaluation of FAZ size in healthy subjects.

Chigger Mite (Acari: Trombiculidae) Survey of Rodents in Shandong Province, Northern China

Chigger mites are parasites of rodents and other vertebrates, invertebrates, and other arthropods, and are the only vectors of scrub typhus, in addition to other zoonoses. Therefore, investigating their distribution, diversity, and seasonal abundance is important for public health. Rodent surveillance was conducted at 6 districts in Shandong Province, northern China (114–112°E, 34–38°N), from January to December 2011. Overall, 225/286 (78.7%) rodents captured were infested with chigger mites. A total of 451 chigger mites were identified as belonging to 5 most commonly collected species and 3 genera in 1 family. Leptotrombidium scutellare and Leptotrombidium intermedia were the most commonly collected chigger mites. L. scutellare (66.2%, 36.7%, and 49.0%) was the most frequently collected chigger mite from Apodemus agrarius, Rattus norvegicus, and Microtus fortis, respectively, whereas L. intermedia (61.5% and 63.2%) was the most frequently collected chigger mite from Cricetulus triton and Mus musculus, respectively. This study demonstrated a relatively high prevalence of chigger mites that varied seasonally in Shandong Province, China.

Evaluation of Six Phenotypic Methods for the Detection of Carbapenemases in Gram-Negative Bacteria With Characterized Resistance Mechanisms

BACKGROUND: We compared the performance of the modified Hodge test (MHT), Triton Hodge test (THT), Carba NP test (CNPt), simplified Carba NP test (CNPt-direct), blue-Carba NP test (BCT), and carbapenem inactivation method (CIM) for rapid and accurate carbapenemase detection. METHODS: The methods were evaluated by using 256 gram-negative isolates, including 197 Enterobacteriaceae (79 Enterobacter spp., 74 Klebsiella spp., 33 Escherichia coli, 10 Citrobacter spp., and 1 Serratia marcescens), 51 Acinetobacter baumannii, and 8 Pseudomonas aeruginosa strains. The collection included 117 non-carbapenemase, 18 Klebsiella pneumoniae carbapenemases (KPC) producers, 46 New Delhi metallo-β-lactamases (NDM) producers, 11 imipenemases (IMP) producers, and 51 oxacillinases (OXA) producers, and 13 strains harboring two different carbapenemase genes. RESULTS: The specificity of the THT (91.5%) was significantly lower than other methods, each of which had 100% specificity (P0.999). Because of improved detection of NDM carriers, THT showed significantly higher sensitivity than the MHT (84.9% vs 75.5%, P<0.001). However, poor performances in detecting OXA still influenced the sensitivities of the CNPt (66.2%) and BCT (82.0%), as well as the MHT and THT. CONCLUSIONS: CNPt-direct and CIM demonstrated the best performance for the efficient detection of carbapenemase among the six evaluated methods. Except the MHT and THT, the detection of carbapenemase-producing Enterobacteriaceae by all the other methods was acceptable, when the OXA-type carbapenemase was not prevalent.

The Changes of Macular Microvasculature and Related Systemic Parameters in Diabetic Patients without Diabetic Retinopathy

PURPOSE: To investigate the systemic parameters related to changes in the retinal microvasculature using optical coherence tomography angiography (OCTA) in diabetic patients without diabetic retinopathy. METHODS: We retrospectively reviewed the medical records of diabetic patients without diabetic retinopathy. All patients showed normal fundus on dilated fundus examinations. OCTA (DRI OCT Triton, Topcon Corporation, Tokyo, Japan) was performed on a 3 × 3 mm area centered at the fovea. The foveal avascular zone (FAZ) area and vascular density (VD) of the superficial and deep capillary plexus (SCP/DCP) were measured. Multiple regression analysis was conducted to evaluate the relationship between microvascular indexes on OCTA and systemic parameters including sex, age, duration of diabetes mellitus (DM), hemoglobin A1c (HbA1c), total cholesterol, presence of hypertension (HTN) and systolic and diastolic blood pressure (SBP/DBP). RESULTS: A total of 35 eyes of 35 patients were included in this study. The mean age was 55.7 ± 11.6 years, the mean duration of DM was 52.4 ± 88.7 months and the mean HbA1c was 8.8 ± 2.3%. The FAZ area of the DCP increased significantly as the HbA1c level increased (p 0.05). No systemic parameters exhibited significant relationships with VD in either the SCP or the DCP (p > 0.05). CONCLUSIONS: The FAZ area of the DCP detected via OCTA increased as the level of HbA1c increased, although the patients did not show diabetic retinopathy on dilated fundus examinations. Therefore, clinicians should consider this when screening diabetic patients for diabetic retinopathy.

Adulthood Benign Triton Tumor Developed in the Orbit

Benign triton tumor (BTT) or neuromuscular hamartoma is an uncommon tumor composed of mature neural and well-differentiated striated muscular elements. Its development is exceptionally rare in the adult and head region. This report describes a case of adulthood BTT that occurred in the orbit. The patient was a 53-year-old woman who presented with right periorbital swelling and pain in eyeball over 2 months. Magnetic resonance imaging revealed a well-enhancing mass surrounding optic nerve and ocular muscles in the right retrobulbar area. The tumor was subtotally removed via transcranial approach. Its pathological diagnosis was confirmed to be a neuromuscular hamartoma. She developed diplopia postoperatively. Adulthood BTT should be considered in the differential diagnosis of head and neck tumors. It is also important to make adequate therapeutic strategy to avoid postoperative neural dysfunction.

Benign Triton Tumor: A Rare Entity in Head and Neck Region

Benign triton tumors (BTT) are very rare lesions composed of mature skeletal muscle and neural tissue. We report a case of a 14-year-old boy who presented with asymptomatic swelling of the chin over an 18-month duration which increased gradually to involve the left side of the lower lip. Clinically, a diagnosis of neurofibroma was made. Excisional biopsy confirmed the diagnosis of a BTT. Having an affinity for large nerve trunks like the brachial and sciatic, these tumors rarely occur in the head and neck region. When they do, they may involve the large central cranial nerve trunk and present as intracranial masses or involve the smaller peripheral nerve twigs and present as asymptomatic skin nodules, of which only four cases involving peripheral nerves are reported in the English literature. Here, we report the fifth documented case of a BTT involving the mental branch of the trigeminal nerve. A brief review of the literature is also provided.

Induction of TNF-alpha, IL-12, IL-10, MCP-1 and IL-8 in Human Peripheral Blood Monocytes in Response to the Cell Wall-Associated Triton X-100 Soluble Protein (TSP) Antigens of Mycobacterium tuberculosis

Mycobacterium tuberculosis is a potent inducer of cytokine production by mononuclear phagocytes, which are an important cellular component in the first line immune defence. In this study, the cell wall-associated Triton X-100 soluble protein (TSP) antigens, TSP-H37Rv, TSP-H37Ra, TSP-K, and TSP-BCG, were isolated from M. tuberculosis H37Rv, M. tuberculosis H37Ra, M. tuberculosis K-strain, and M. bovis BCG, respectively. The monocytes were isolated from the peripheral blood mononuclear cells of healthy individuals and were co-cultured with each TSP antigens and the secretory proteins of M. tuberculosis (PPD and 30-kDa antigen) to measure the production of cytokines; tumor necrosis factor (TNF)-a, interleukin (IL)-12, IL-8 and monocyte chemotactic protein-1 (MCP-1). The TSP-H37Rv antigen- stimulated monocytes showed higher level of TNF-a and IL-12 production compared to those of other TSP antigens and PPD. Especially, IL-12 production in response to the TSP-H37Rv antigen was significantly elevated in comparison with that of PPD-stimulated monocytes (TSP-H37Rv, 255.5+/-256.9 pg/ml; PPD, 55.7+/-55.4 pg/ml). However, the 30-kDa antigen did not induce TNF-alpha expression and also showed the lowest level of cytokine and chemokine production by monocytes. MCP-1 and IL-8 production were similarly increased in response to all TSP antigens and the PPD antigen. The production of IL-12 by the TSP-H37Rv antigen stimulation was significantly increased in PPD reactors than that in the non-reactor group, while the levels of other cytokines stimulated with each TSP antigens, 30-kDa and PPD antigen were not significantly different between the tuberculin reactor and the non-reactor groups. These results suggest that the cell wall-associated TSP antigen isolated from M. tuberculosis H37Rv acts as a more potent IL-12 inducer than the PPD antigen in innate immune response and thus it could further activate the Th1-mediated immune responses effectively against M. tuberculosis infection.

Mycobacteria-induced Interleukin-12 Expression by Human Monocyte-derived Macrophages Is Negatively Regulated by Phosphatidylinositol 3-kinase and ERK 1/2 Pathways

Both interleukin (IL)-12, an important cytokine skewing the immune response towards a Th1 cytokine profiles, and tumor necrosis factor (TNF)-alpha, are thought to be critical factors in defenses against mycobacteria. In this study, we evaluated the roles of phosphatidylinositol 3-kinase (PI 3-K), and extracellular signal-regulated kinase (ERK) 1/2 pathways in the expression of IL-12 in human monocyte-derived macrophages (MDMs) after stimulation with Mycobacterium tuberculosis H37Rv (M. tbc) or the Triton X-114 solublized proteins (TSP) of M. tbc. Both M. tbc and TSP rapidly phosphorylated ERK 1/2, and Akt in human MDMs. Inhibition of PI 3-K-Akt pathway by specific inhibitors (LY294002 and wortmannin) dramatically increased M. tbc- or TSP-induced IL-12 p40 and p35 mRNA and IL-12 production. In addition, blockade of ERK 1/2 pathway by specific inhibitors (PD98059 and U0126) significantly increased the mRNA levels and cytokine production in M. tbc- or TSP-treated MDMs. On the contrary, M. tbc- or TSP-induced TNF-a production was significantly depressed in human MDMs by pretreatment with inhibitors of PI 3-K or ERK pathways. The M. tbc or TSP stimulation decreased ERK 1/2 phosphorylation by 70% in the presence of wortmannin or LY294002, suggesting that some cross-talk between the PI 3-K-Akt and mitogen-activated protein kinase kinase (MEK)-ERK pathways may be operating in human monocytes during mycobacterial infection. PI 3-K activity is partially required for the M. tbc- or TSP-induced ERK 1/2 phosphorylation. Collectively, these data suggest that the PI 3-K and ERK 1/2 pathways play a central role in the negative regulation of IL-12, but not TNF-a, production by M. tbc.

Isolation of a Specific Antigen from the Triton X-100 Soluble Protein (TSP) Of Mycobacterium tuberculosis

Tremendous efforts have been made to develop better vaccines and diagnostic markers for the effective control of tuberculosis. Recently, we reported that the Triton X-100 soluble protein (TSP) of Mycobacterium tuberculosis induced strong T-cell proliferation and IFN-gamma production in humans, and also conferred a significant level of protection against tuberculosis in a mouse model. In this study, the TSP was prepared by Triton X-100 extraction of Mycobacterium tuberculosis bacilli, which was followed by Triton X-114 phase partitioning. Western blot analysis using sera of 177 active pulmonary tuberculosis patients and 323 healthy individuals revealed that the TSP contained a immunodominant 40-kDa antigen specifically reacting with some sera from pulmonary tuberculosis patients. The 40-kDa antigen was purified by ion-exchange chromatography, and partially characterized by two-dimensional gel electrophoresis and N-terminal sequencing. Results of this study suggest that 40-kDa molecule of the TSP antigen from the cell suface of Mycobacterium tuberculosis can be used as a serodiagnostic marker as well as a potential vaccine candidate against tuberculosis.

Identification of putative pathogens in acute endodontic infections by PCR based on 16S rDNA / 대한치과보존학회지

The purpose of this study was to investigate the frequency of 7 putative pathogens in endodontic infections. The specimens were collected from infected pulpal tissue of patients who were referred for root canal treatment to the department of conservative dentistry, Chosun University. Samples were collected aseptically using a barbed broach and a paper point. The cut barbed broaches and paper points were transferred to an eppendorf tube containing 500 ml of 1 X PBS. DNAs were extracted from the samples by direct DNA extraction method using lysis buffer (0.5% EDTA, 1% Triton X-100). Identification of 7 putative pathogens was performed by PCR based on 16S rDNA. The target species were as follows: Porphyromonas endodontalis, Porphyromonas gingivalis, Prevotella intermedia, Prevotella nigrescens, Bacteroides forsythus, Actinobacillus actinomycetemcomitans, and Treponema denticola. Our data revealed that the prevalence of P. endodontalis was found in 88.6% (39/54), P. gingivalis 52.3% (23/44), P. nigrescens 18.2% (8/44), P. intermedia 15.9% (7/44), B. forsythus 18.2% (8/44), A. actinomycetemcomitans 2.3% (1/44), T. denticola 25% (11/44) of the samples. The high prevalence of P. endodontalis and P. gingivalis suggests that they may play an important role in the etiology of endodontic infections.

Epidemilogical Study on Orientia tsutsugamushi Infection of Wild Rodents in Korea

This study was carried out to determine the seropositivity for Orientia tsutsugamushi infection among wild rodents captured in farms and mountains of Korea. A total 965 wild rodents of 7 species and 48 Crocidura laciura were captured in farms and mountains of 6 province during 1997~2000 in Korea. Serological evidence for O. tsutsugamushi infection was obtained using O. tsutsugamushi antigens by indirect immunofluorescent antibody technique (IFA). Four hundred eighty four Apodemus agrariusus, 246 Apodemus peninsulae, 179 Eothenomys regulus, 44 Microtus fortis, 6 Micromys minutus, 5 Mus musculus, and 1 Cricetula triton were captured in Gyeonggi, Gangwon, Chungnam, Gyeongbuk, Gyeongnam, and Jeonbuk province. Among them, 59 (12.2%) A. agrariusus, 22 (8.9%) A. peninsulae, 12 (6.7%) E. regulus, 5 (11.4%) M. fortis and 1 (16.7%) M. minutus were IF antibody positive against O. tsutsugamushi Gilliam or Karp strain.

Measurement of specific IgE to abalone(Haliotis discus hannai) and identification of IgE binding components / 천식및알레르기

BACKGROUND: Abalone, which is a member of the shellfish family, can often induce severe allergic reactions in sensitized individuals, but there have been only a few studies ofn its allergenic components. A recent study has identified two major allergens with molecular weights of 38 and 49 kDa in South African abalone. OBJECTIVES: The aim of this study is to evaluate skin test prevalence and IgE sensitization to northern disk abalone (Haliotis discus hannai) which is one of the major abalones in this country, and to identify its allergenic components. METHODS: Skin prick tests were performed with 62 home-made extracts of domestic foods including abalone, turban shell, triton shell, shrimp etc. in 1,738 patients with various allergic diseases. Serum specific IgE antibodies to abalone were determined by ELISA in 81 positive responders on skin prick tests to abalone extract and 40 non-atopic healthy controls. ELISA inhibition tests were performed to evaluate cross-allergenecity between abalone and other sea foods(turban shell, triton shell, shrimp and house dust mite). Allergenic components of Haliotis discus hannai were identified by SDS-PAGE and IgE-immunoblot analysis. RESULTS: The positive response rate(A/H ratio> or=2+) to abalone on skin prick test was 4.7% in patients with various allergic diseases. Serum spcecific IgE to abalone was detected in 23(34.5%) of 67 patients. Serum specific IgE levels to abalone tended to increase according to skin test reactivity without statistical significance(p>0.05). ELISA inhibition tests showed significant dose-dependent inhibitions with addition of turban shell, triton shell and shrimp extracts IgE immunoblot analysis showed ten allergenic components (33, 37, 40, 60, 63, 71, 76, 86, 92, 111 kDa), of which seven allergens (40, 60, 63, 71, 76, 86, 92 kd) were bound to IgE in more than 50% of the sera tested. CONCLUSION: The sensitization rate to abalone was 4.7% in allergy patients. Serum specific IgE to abalone was detected by ELISA, and 7 major allergens within abalone were identified. Further studies will be needed to elucidate their clinical significances.

Specific IgE determination to shrimp (Metapenaeus joyneri) and identification of its allergen / 대한내과학회지

BACKGROUND: Shrimp is one of the major causative crustacean food allergen. An investigation has been reported that tropomyosin belonged to muscle protein is a major allergen within shrimp. But there have been a few investigations on shrimp allergen in Korea. The aim of this study is to evaluate skin reactivity and specific IgE sensitization to Metapenaeus joyneri which is one of the major shrimp in this country, and to identitify IgE binding components and evaluate allergenic relationship with other species. METHODS: We performed skin prick test with M. joyneri extract in 1,738 patients. ELISA was performed for detection of serum specific IgE antibody. To evaluate the cross allergenecity between M. joyneri and other crustaceans (crab, lobster, crayfish), Dermatophagoides pteronyssinus (Dpt), triton shell, abalone and buckwheat. ELISA inhibition tests were performed with each four patient's sera showing high specific IgE antibody. To identify IgE binding components, SDS-PAGE followed by IgE-Immunoblot were applied. RESULTS: 211 patients (12.2%) showed positive responses (A/H >or=2+) on skin prick test. Serum specific IgE antibodies were detected in 61 patients (37.2%) of 164 sensitzed patients. ELISA inhibition test using four patient's sera showed significant inhibitions by M. joyneri. and other crustaceans including lobster, crab and crayfish, partial inhibitions were noted by Dpt, triton shell, buckwheat and abalone. SDS-PAGE and IgE-imunoblot with patients' individual sera sensitized to M. joyneri showed 12 IgE binding components (31, 32, 38-44, 57, 70, 81 kDa) and two (31, 32 kDa) were bound to IgE in more than 50% of sera tested. Five (43, 44, 57, 70 and 81 kDa) were bound to IgE in more than 25% of sera tested. CONCLUSION: Specific IgE was detected in 37.2% of allergy patients sensitized to M. joyneri. Twelve IgE binding components and two (31, 32 kDa) major allergens were indentified. Cross allergenecity was noted with other crustaceans.

Subcutaneous Neuromuscular Hamartoma: A case report

Subcutaneous form of neuromuscular hamartoma is extremely rare and histologically different from the conventional neuromuscular hamartoma of the peripheral nerve or benign Triton tumor by an indistinct nodular growth with ill-defined margin and marked collagen interposition. It is usually not associated with a major nerve. We report a case of subcutaneous neuromuscular hamartoma developed in the forehead of 24-year-old man. The tumor showed proliferation of dense, hyalinized fibrous tissue, in which single or group of mature skeletal muscle fibers and nerve fibers were haphazardly intermixed. Recognition of abnormally arranged muscle and nerve fibers is important not to miss this lesion.

A Case of Malignant Triton Tumor In Nasal Cavity / 대한이비인후과학회지

Malignant triton tumor is a rare and usually aggressive sarcoma consisting of a malignant schwannoma with rhabdomyoblastic differentiation. Although this tumor is usually found at the extremities, 20% of cases are located in the head and neck region. This unusual tumor has not yet been described in the Korean Journal of Otolaryngology-Head and Neck Surgery. Malignant triton tumor is histologically high-grade, with large numbers of mitoses, prominent necrosis, and cellular pleomorphism. The clinical course for malignant triton tumor is usually one of rapid growth, early metastasis, and poor outcome in spite of therapy. Recently we experienced a case of malignant triton tumor, which involved the nasal cavity, maxillary and ethmoid sinus. The malignant tumor was treated with wide surgical excision followed by radiation therapy.

Human Cellular Immune Responses to the Aqueous Fraction of the TSP Antigen of Mycobacterium tuberculosis H37Rv

Phase-partitioning with Triton X-114 (TX114) was applied to the TSP antigen, which may be preferentially associated with the cell wall of M. tuberculosis. The hydrophilic protein components of the TSP antigen were successfully separated from integral hydrophobic macromolecules. To further characterize and examine the cellular immune response of the aqueous fraction of the TSP antigen (TSPa), the in vitro properties of the antigen were measured by lymphoproliferation; surface expression of IL-2 Ra on T lymphocytes was analyzed by flow cytometry; and the cytokine mRNA expression pattern was determined by RT-PCR. Significant lymphoproliferative responses to the TSPa antigen were observed in healthy tuberculin reactive donors after a 5 day in vitro stimulation. TSPa treatment of PBMCs from healthy tuberculin positive subjects for 5 days resulted in progressive augmentation of IFN-r, II 2, and IL-2Ra mRNA expression, as measured by RT-PCR, but considerably reduced IL-4 mRNA expression. In addition, the TSPa antigen stimulated more IL-12 p40 mRNA production than did the PPD antigen, and graduaBy suppressed IL- 10 mRNA expression. Moreover, the CD3' T cells of tuberculin positive subjects displayed a profound increase in their expression of the II 2Ru protein (39.0%) in response to the TSPa antigen. Proliferation was correlated with IL-2 and IL-2Ra mRNAs, but not correlated with distinct IFN-r or IL-12 p40 mRNA production. These findings strongly suggest that the TSPa antigen preferentially evokes the generation of a Thl-like immune response in healthy tuberculin reactors.

Activation of Colon Cancer-Draining Lymph Node Mononuclear Cells by 30 kDa or Triton X-100 Solubilized Protein Antigen of Mycobacterium tuberculosis

Tumor-draining lymph node mononuclear (TDLMN) cells are specifically sensitized to the growing tumor but such cells are deficient for mediating an antitumor response. In this study, we examined the feasibility of using mycobacterial 30 kDa or Triton X-100 solubilized protein (TSP) antigen to stimulate mononuclear cells of colon cancer-draining lymph node for the generation of cell mediated immune effector cells. The proliferative response of TDLMN cells stimulated with mycobacterial 30 kDa or TSP antigen was determined by H-thymidine incorporation assay. The proliferation of TDLMN cells to mycobacterial 30 kDa or TSP antigen was significantly increased in PPD (+) patients, but a poor response to the 30 kDa or TSP antigen was observed in PPD (-). The expression on ro T cells to mycobacterial 30 kDa or TSP antigen was assessed by flow cytometry. The ro T cells from PPD (+) patient responded only to 30 kDa antigen but to TSP antigen. An investigation of cytokine mRNA expression was undertaken using reverse transcription- polymerase chain reaction (RT-PCR) to follow TDLMN cells stimulated with the 30 kDa or TSP antigens for 5 days. The IFN-r and TNF-a mRNA expression was only induced in TDLMN cells of PPD (+) patient in response to the 30 kDa or TSP antigen. The IL-2 mRNA expression was induced in both PPD (+) and PPD (-) in response to the 30 kDa or TSP antigen. But the IL-4 mRNA expression was not induced in response to the 30 kDa or TSP antigen. These results suggest that the 30 kDa and TSP antigens may serve as biologic response modifier for the generation of cell mediated immune effector cells.

Determination of Anti-tuberculous Antibody Against Triton X-100 Solubilized Protein ( TSP ) Antigen of Mycobacterium tuberculosis in the Sera of Patients with Pulmonary Tuberculosis

Some of the proteins of mycobacteria are preferentially associated with the cell wall and are powerful immunogens, and humoral antibody responses to these mycobacterial antigens may occur in patients with tuberculosis. In this study, Triton X-100 solubilized protein (TSP) antigen was isolated from Mycobacterium tuberculosis H37Rv by overnight shaking with 1% Triton X- 100/PMSF and 10-90% ammonium sulfate precipitation. IgG and IgM antibody levels against TSP, crude protein from the unheated cultrue filtrate (CF#) and 30 kDa antigens were determined in the sera of 80 patients with pulmonary tuberculosis and 99 healthy controls with PPD (+) and (-). High IgG reactivity to TSP and CF antigen was observed in tuberculosis patients. Mean IgG antibody titers against all of three mycobacterial antigens were differed significantly (P<0.01) between patients and controls but IgM showed no difference. By the cut-off value adding 2 standard deviation to the mean absorbance of controls, the sensitivity and specificity of the IgG antibody to TSP antigen were 93.9% and 77.5%. The specificity to TSP antigen was a litttle higher than those obtained by CF and 30 kDa antigen. From the above results, the TSP antigen may be useful for the serodiagnosis of tuberculosis.