Induction of immunological tolerance by oral, but not intravenous and intraportal, administration of ovalbumin and the difference between young and old mice.

BACKGROUND: Induction of immunological tolerance is dependent on the route of antigenic administration, the dose of an antigen and the age of animals. OBJECTIVES: We investigated the effect of age on the tolerance induction in mice by administration of antigen through different routes and at different doses. DESIGN: Young and old BDF1 mice were orally, intraportally or intravenously administrated with a low or a high dose of ovalbumin (OVA). Then, delayed-type hypersensitivity (DTH) responses and serum anti-OVA antibody levels were assessed after systemic immunization of OVA with alum after appropriate intervals. RESULTS: In the young mice, oral administration of OVA suppressed DTH response and anti-OVA IgG1, IgG2b, IgM and IgE level in a dose-dependent manner. In the old mice, however, the suppression of IgG1 and IgE levels was induced by oral administration of a low dose of OVA, but no suppression by a high dose. On the other hand, intraportal or intravenous injection of OVA did not suppress DTH response and enhanced anti-OVA antibody levels in a dose-dependent manner in both young and old mice. Production of anti-OVA IgG2a antibody after systemic injection of OVA was detected in the mice, which had been treated with intraportal or intravenous injection of OVA, but not detected in the mice, which had been treated with oral administration of OVA. On the contrary, suppression of anti-OVA IgE antibody was observed only in the mice, which had been treated with oral administration of OVA. CONCLUSION: The oral administration of OVA, neither intravenous nor intraportal, induced immunological tolerance to OVA. An adequate dose of OVA for the tolerance induction and the suppression of antibody production are different between young and old mice. The suppression of IgE antibody was observed only by oral administration of OVA, much obviously in young mice than in the old. The results also indicated that the antigen processing in the liver did not play a major role in the induction of oral tolerance to OVA.

Predicting Autism Spectrum Quotient (AQ) from the Systemizing Quotient-Revised (SQ-R) and Empathy Quotient (EQ).

BACKGROUND: Empathizing is a specific component of social cognition. Empathizing is also specifically impaired in autism spectrum condition (ASC). These are two dimensions, measurable using the Empathy Quotient (EQ) and the Autism Spectrum Quotient (AQ). ASC also involves strong systemizing, a dimension measured using the Systemizing Quotient (SQ). The present study examined the relationship between the EQ, AQ and SQ. The EQ and SQ have been used previously to test for sex differences in 5 'brain types' (Types S, E, B and extremes of Type S or E). Finally, people with ASC have been conceptualized as an extreme of the male brain. METHOD: We revised the SQ to avoid a traditionalist bias, thus producing the SQ-Revised (SQ-R). AQ and EQ were not modified. All 3 were administered online. SAMPLE: Students (723 males, 1038 females) were compared to a group of adults with ASC group (69 males, 56 females). AIMS: (1) To report scores from the SQ-R. (2) To test for SQ-R differences among students in the sciences vs. humanities. (3) To test if AQ can be predicted from EQ and SQ-R scores. (4) To test for sex differences on each of these in a typical sample, and for the absence of a sex difference in a sample with ASC if both males and females with ASC are hyper-masculinized. (5) To report percentages of males, females and people with an ASC who show each brain type. RESULTS: AQ score was successfully predicted from EQ and SQ-R scores. In the typical group, males scored significantly higher than females on the AQ and SQ-R, and lower on the EQ. The ASC group scored higher than sex-matched controls on the SQ-R, and showed no sex differences on any of the 3 measures. More than twice as many typical males as females were Type S, and more than twice as many typical females as males were Type E. The majority of adults with ASC were Extreme Type S, compared to 5% of typical males and 0.9% of typical females. The EQ had a weak negative correlation with the SQ-R. DISCUSSION: Empathizing is largely but not completely independent of systemizing. The weak but significant negative correlation may indicate a trade-off between them. ASC involves impaired empathizing alongside intact or superior systemizing. Future work should investigate the biological basis of these dimensions, and the small trade-off between them.

Synthesis of cyclopentadienones catalyzed by methylidynetricobalt nonacarbonyl.

Easily prepared and air-stable methylidynetricobalt nonacarbonyl could be used as a catalyst for the intramolecular [2 + 2 + 1]-cocyclization of diynes and carbon monoxide producing cyclopentadienones.

The effects of allopurinol on immune function in normal BALB/c and SCID mice.

To clarify the relationship between purine metabolism and immunity, the in vivo immunosuppressive effects of allopurinol (AL), a xanthinoxidase (XO) inhibitor, were studied using normal BALB/c and severe combined immunodeficient (SCID) mice. Following AL administration for 14 weeks (long term), a decreased immune response to ovalbumin (OVA) in the peripheral blood was observed in normal mice, which might not be only due to direct B cell suppression but also due to suppression of helper T cell function. In the SCID mice, there was a markedly late and reduced recovery of surface immunoglobulin (sIg) positive cells, which are markers for mature B lymphocytes, in the peripheral blood after AL administration. The total immunoglobulin G (IgG) titers in the AL treated group were significantly lower than in the control group 6 weeks after stem cell transfer, but increased until there was no difference in the titers between the two groups at week 14. CD4 positive helper T cells and CD8 positive T cells were slow to recover, though these gradually recovered to reach normal levels in the mature stage. These data suggest that the administration of AL may modulate B cell and T cell responses in OVA-immunized antibody formation. Furthermore, this study showed that AL could influence immune functions during the pre-natal and developmental periods and that its effects might differ according to the stages of maturity of the immune cells.

Differential age effect of oral administration of an antigen on antibody response: an induction of tolerance in young mice but enhancement of immune response in old mice.

Sheep red blood cells (SRBC) were orally administered to young (4 months old) and old (22 months old) mice, and its effect on the antibody production after systemic immunization was compared between young and old mice. The results showed that the dose-dependent suppression of antibody response (oral tolerance) was observed in young mice which had been previously treated with oral administration of SRBC. On the contrary, the enhancement of antibody production was observed in old mice which had been treated in the same way. The enhanced level of IgG antibody in old mice was higher than that of young mice. The critical age determining either suppression or enhancement of antibody response after the oral administration of the antigen was present between 6.5 and 10.5 months of age. When the oral administration of the antigen was performed in young (3 months old) and middle-aged mice (12 months old), the oral tolerance for young and the enhanced antibody response for middle-aged mice were observed even at 6 months after the treatment. The analysis by in vitro antibody response using T and B cells prepared from young and old mice showed that age-related alteration of T and B cells is responsible for the suppression and the enhancement of antibody response after oral administration of SRBC, respectively.

Age-related alteration of cytokine production profile by T cell subsets in mice: a flow cytometric study.

Spleen cells from young and old C57BL/6 mice were stimulated with a combination of anti-CD3 and anti-CD28 antibodies, and the profile of cytokine production was examined by two different methods; the concentrations of cytokines as measured by ELISA, and identification of cytokine-positive cells by flow cytometry. The ELISA method revealed that IL-2 production by spleen cells after stimulation was significantly lower in the old mice compared to the young mice. while IFN-gamma production was the reverse. The flow cytometric analysis showed that the percentage of IL-2 positive cells in spleen cells after the stimulation was significantly lower in the older mice than in the young mice, and vice versa for the percentage of IFN-gamma-positive cells. Regarding the T cell subsets, CD4+ T cells were a major source of IL-2 in both the young and old mice. IL-2-positive cells in both CD4+ and CD8+ T cells showed a significant decrease with age. On the contrary, CDX T cells were the major source of IFN-gamma. An age-related increase of IFN-gamma positive cells was observed in both CD4+ and CD8+ T cells. CD4 T cells were the major source of IL-4, and the percentage of IL-4-positive CD4+ T cells also increased with age, although the level of IL-4 production was modest in C57BL/6 mice compared with IL-2 and IFN-gamma. Such age-related changes of cytokine production are presumed to play an important role in the alteration of immunological capacity with age.

Role of inhibition of penicillin binding proteins and cell wall cross-linking by beta-lactam antibiotics in low- and high-level methicillin resistance of Staphylococcus aureus.

We have investigated the relationships between the minimum inhibitory concentrations (MICs) of eight beta-lactams and their binding affinities of penicillin binding proteins in low- and high-level methicillin-resistant Staphylococcus aureus (MRSA). For high-level MRSA (H-MRSA), a significant correlation was found between IC50 for PBP2A and MIC, whilst no clear relationship was apparent for low-level MRSA (L-MRSA). However, the product of the IC50s for PBP2A and PBP4 significantly correlated with the MIC in L-MRSA. Since PBP4 is thought to mediate secondary cross-linking of the staphylococcal cell wall, we analyzed the effect on cell wall cross-linking of L- and H-MRSA of two representative drugs (cefoselis and flomoxef). Interestingly, the decrease in cell wall cross-linking, which was clearly observed in L-MRSA, was diminished with H-MRSA. It is concluded that for L-MRSA, a reduction in cell wall cross-linking caused by inhibiton of PBP4 may contribute to the antimicrobial activity of beta-lactam antibiotics, while for H-MRSA, inhibition of PBP2A mainly determines the antimicrobial activity. Since neither alteration of expression nor binding affinity of PBP4 were observed in these studies, unknown factors must operate to diminish the effect of PBP4 inhibition and contribute to the mechanism of high level resistance of MRSA.

Vitamin E enhances the immune functions of young but not old mice under restraint stress.

Young and old C57BL/6 male mice were given a diet containing a high dose of vitamin E (VE treatment) and its effect on the immune system was examined before and after the exposure to restraint stress. The VE treatment per se gave rise to a slight increase of splenic T cells in percentage and a significant enhancement of Con A response of spleen cells in young, but not in old mice. The VE treatment also resulted in the enhancement of production of IL-2 and IFNgamma in young, but not in old mice. Restraint stress led to thymic involution in both young and old mice. This thymic involution was not ameliorated by the VE treatment. Percentage of splenic T cells and their mitogenic response decreased just after the stress, but soon rebounded over the control level. The VE treatment further enhanced the recovery after the stress in young mice, but on the contrary suppressed the recovery in old mice. The results in the present study suggested that the VE treatment was effective in the prevention of immunological decline of young mice before and after the exposure to the stress. On the other hand, such a preventive effect was not observed in old mice that were already in the depressed state of immunological functions.

Relationship between apolipoprotein E polymorphism, menopausal symptoms, and serum lipids during hormone replacement therapy.

OBJECTIVE: Menopausal women receive hormone replacement therapy (HRT) to relieve symptoms and to help prevent osteoporosis or atherosclerotic disease. We investigated the association of apolipoprotein (apo) E polymorphism with menopausal symptoms, body fat mass and lipid profile in 236 women, together with the lipid changes accompanying HRT administration in 172 women from this population who were postmenopausal. STUDY DESIGN: The subjects were divided into three groups according to apo E phenotype: group E2, apo E2/2 and 2/3; group E3, apo E3/3; group E4, apo E4/3 and 4/4. Typical menopausal symptoms were classified into four degrees of severity; body fat mass, lipid profile, and serum lipid levels were measured before and 6 months after oral HRT. RESULTS: There were no significant differences between the symptoms of the three groups. The serum levels of apo E were the highest in group E2 and lowest in group E4. Analogous tendencies were seen in the mean levels of total cholesterol (TC), low density lipoprotein cholesterol (LDL-C), and apo B, with group E4 having the highest levels and group E2 the lowest. Triglyceride levels (TG) were the highest in group E2, but the difference was not significant. These parameters suggest that group E4 had the highest risk of cardiovascular disease. The LDL-C/high density lipoprotein cholesterol (HDL-C) ratio was improved from 2.18 before HRT to 1.52 after HRT in group E2; from 2.26 to 1.92 in group E3; and from 2.57 to 2.10 in group E4. CONCLUSION: The apo E phenotype was not associated with any difference in menopausal symptoms. Group E4 had the highest risk for cardiovascular disease, and group E2 the lowest. Oral HRT could be recommended for the women in group E4.

Biochemical and pharmacological characterization of FK706, a novel elastase inhibitor.

FK706, sodium 2-[4-[[(S)-1-[[(S)-2-[[(RS)-3, 3, 3-trifluoro-1-isopropyl-2-oxopropyl]aminocarbonyl]pyrrolidin -1-yl]carbonyl]-2-methylpropyl] aminocarbonyl] benzoylamino] acetate, C26H32F3N4NaO7, is a synthetic water-soluble inhibitor of human neutrophil elastase. This compound demonstrated a competitive and slow-binding inhibition of human neutrophil elastase with a Ki of 4.2 nM. In studies using synthetic substrates, FK706 inhibited human neutrophil elastase activity and porcine pancreatic elastase activity with respective IC50 values of 83 and 100 nM. FK706, however, inhibited more weakly, (IC50 values > 340 microM) other serine proteinases such as human pancreatic alpha-chymotrypsin, human pancreatic trypsin and human leukocyte cathepsin G. FK706 also effectively inhibited the hydrolysis of bovine neck ligament elastin (2 mg/ml final concentration) by human neutrophil elastase (4 microg/ml final concentration) with an IC50 value of 230 nM. FK706 protected animals against human neutrophil elastase (50 microg/animal)-induced lung hemorrhage with ED50 values of 2.4 microg/animal by intratracheal administration and 36.5 mg/kg by intravenous administration, respectively. Subcutaneous administration of FK706 significantly suppressed human neutrophil elastase (20 microg/paw)-induced paw edema in mice in a dose-dependent manner (47% inhibition at a dose of 100 mg/kg). These results suggest that FK706 would be a useful tool for investigating the role of human neutrophil elastase in inflammatory disorders associated with an excess of elastase, such as pulmonary emphysema, adult respiratory distress syndrome, septic shock, cystic fibrosis, chronic bronchitis and rheumatoid arthritis.

Regulation of experimental autoimmune orchitis by the presence or absence of testicular antigens during immunological development in SCID mice reconstituted with fetal liver cells.

Severe combined immunodeficient (SCID) mice were immunologically reconstituted by the transfer of fetal liver cells (FLC) of BALB/c mice (SCID-FLC mice). In peripheral blood (PB) of SCID-FLC mice, B and T cells started to appear 2 and 5 weeks, respectively, after the transfer of FLC, and had attained normal levels by 7 weeks. Orchidectomy and transplantation of testis under the kidney capsule were conducted at various stages of immunological maturation, and the induction of experimental autoimmune orchitis (EAO) was performed after immunological maturation in SCID-FLC mice. The experimental system was used to establish that the presence of testicular antigens in the early stage of immunological development influences the induction of EAO; grade of EAO was reduced in the presence of the antigens, and enhanced in their absence. In other words, the existence of self tissue antigens in the early stage of immunological development was essential for proper establishment of tolerance to the self tissue. These findings suggested that the SCID-FLC mouse is a suitable model with which to analyse the interaction between self antigens and cells of the developing immune system, which is otherwise observed only in the fetal or perinatal stage in experimental animals.

Development of the immune system in severe combined immunodeficiency mice reconstituted with transferred fetal liver cells.

Severe combined immunodeficiency (SCID) mice were immunologically reconstituted by the transfer of fetal liver cells (FLC) from BALB/c (SCID-isoFLC mice) or C57BL/6 mice (SCID-alloFLC mice). The developmental process of the immune system in the peripheral blood (PB) was almost comparable between SCID-isoFLC and SCID-isoFLC mice. Analysis of the lymphoid organs and the PB from SCID-alloFLC mice indicated that slg+ B cells appeared and were distributed to the periphery within 2 weeks after the transfer of FLC. It was also suggested that precursor T cells entered the thymus before 4 weeks after the transfer, and were differentiated into mature CD4+ or CD8+ T cells which then migrated to the periphery by 6 weeks. All of mature lymphocytes in the periphery of the SCID-alloFLC mice were shown to express donor-type H-2 antigens. Additionally, in the SCID-isoFLC mice, cell-mediated immunity such as rejection against alloantigens was functioning from 6 weeks, and humoral immune function was suggested by the detection of cells generating antigen-specific antibodies. We discuss that development of the immune system in SCID mice receiving transferred FLC was comparable to that normally seen in the fetal and neonatal stages.

Anatomic segmental resection of the lung by thoracoscopy: an experimental study.

In patients who are unable to undergo a lobectomy for a small peripheral lung cancer, a partial thoracoscopic resection appears to be one viable alternative. However, since the regional lymphatics are disrupted in an anatomical fashion with a segmentectomy, it appears superior to a wedge resection. This experimental study was conducted to determine whether or not an anatomical segmental resection is feasible by thoracoscopy. A segmental resection of porcine lungs was performed using thoracoscopy. The segmental vessels were divided between ligatures. The segmental bronchus was divided by an endoscopic stapler. The intersegmental lung parenchyma was divided using a cotton dissector and a contact neodymium-yttrium aluminum garnet laser. Forty-three pigs were divided into seven groups as follows. Group 1: S1 + 2; group 2: S3; group 3: upper division; group 4: lower division; group 5: S6; group 6:S8; and group 7: S9 + 10. The operating times ranged from 145 +/- 15 min to 191 +/- 47 min. Blood loss ranged from 36 +/- 35 ml to 151 +/- 48 ml in all groups. The blood loss in the group with a resection of S6 and S9 + 10 was significantly greater than that of the other five groups. Most of the blood loss occurred during the division between the intersegmental planes. In conclusion, a thoracoscopic segmentectomy is considered to be technically feasible; however, further refinements in this technique are warranted before beginning clinical trials.

Thoracoscopic en bloc total esophagectomy with radical mediastinal lymphadenectomy.

OBJECTIVE: Total esophagectomy with en bloc mediastinal lymphadenectomy for cancer carries a substantial morbidity and mortality rate. To investigate the feasibility of thoracoscopic technique, we carried out an extensive laboratory study. Encouraged by our excellent results, we conducted a clinical trial. METHODS: From September 1994 to September 1995, 39 patients thoracic esophageal cancer lesions not invading surrounding organs underwent total esophagectomy with mediastinal lymphadenectomy by means of thoracoscopy. Ages ranged from 47 to 86 years. The procedures were conventional except for the thoracic portion, which was performed as a thoracoscopic procedure with six trocar holes instead of thoracotomy. All harvested lymph nodes were counted for each station. Spirometric data and plethysmographically determined vital capacity were measured before and after operation for all patients. RESULTS: All procedures were accomplished as scheduled, and none was converted to open thoracotomy. The operating time was 200 +/- 41 minutes (mean +/- standard deviation). Estimated blood loss was 270 +/- 157 ml. The harvested lymph nodes numbered 19.7 +/- 11.1 per patient. Seventeen patients (45%) had positive lymph nodes. There were no in-hospital deaths within 30 days. Twenty-two patients did not require postoperative ventilatory support. Vital capacity decreased to 85% +/- 11% of the preoperative values, and forced expiratory volume in 1 second decreased to 82% +/- 16%. CONCLUSIONS: Thoracoscopic mediastinal lymphadenectomy is technically feasible, and its completeness is comparable to that of the open technique. The decline in pulmonary function is significantly less than that seen in our previous experience with the open technique.

Prevalence of severity of hypoxemia following clinical voluntary hyperventilation.

The voluntary hyperventilation (VHV) test is used in many clinical examinations. However, arterial hypoxemia following a clinical VHV test is not a well-studied phenomenon. We analyzed the arterial blood gases (ABGs) of 61 patients during a VHV test. The ABG were taken prior to (PaO2-Prior), immediately following (PaO2-Immediate), and 5 min after (PaO2-After) the VHV test. The patients' average PaO2 rose significantly (p < 0.0001) from the PaO2-Prior (88 +/- 8 mm Hg; mean +/- SD) to the PaO2-Immediate (118 +/- 13 mm Hg) and then dropped significantly (p < 0.0001) to the PaO2-After (74 +/- 16 mm Hg). Two of the 20 patients who experienced an angina pectoris attack (AP(+)) following the VHV test showed severe arterial hypoxemia (PaO2-After < 60 mm Hg), whereas 9 of the 41 patients who did not experience an angina pectoris attack (AP(-)) showed a PaO2-After < 60 mm Hg. The PaO2-After did not correlate with the PaO2-Prior. The decrease in the PaO2-Prior to After did not correlate significantly with the left ventricular ejection fraction rate (n = 58, r = 0.18, not significant). However, the decrease in the PaO2-Prior to After correlated well with the degree of recovery of the PaCO2 following the VHV test (r = -0.69, p < 0.0001). The age, gender ratio, changes in arterial blood gases, number of patients who experienced PaO2-After < 60 mm Hg, and left-ventricular ejection fraction rate were not significantly different between the AP(-) and AP(+) groups. Posthyperventilation hypoxemia developed frequently following the VHV test during coronary angiography. Although this arterial hypoxemia was not directly correlated with the occurrence of AP attacks following VHV in this study, continuous SaO2 monitoring is recommended whenever a VHV test is used as a diagnostic technique to avoid the potentially deleterious effects of arterial hypoxemia.

Unilateral thoracoscopic laser pneumoplasty of diffuse bullous emphysema.

The technical details of thoracoscopic laser pneumoplasty (TLP) by means of a contact neodymium:yttrium-aluminum-garnet (Nd:YAG) laser is described. Retrospective analysis of 769 unilateral TLPs indicated that the operative risk is acceptable (6% 3-months mortality rate), and significant subjective and objective improvement was achieved. Recurrent bullous emphysema is discussed. TLP is an effective treatment of advanced emphysema.

Thoracoscopic laser pneumoplasty in the treatment of diffuse bullous emphysema.

BACKGROUND: Thoracoscopic laser pneumoplasty in the treatment of diffuse bullous emphysema by means of a contact neodymium:yttrium-aluminum garnet laser was evaluated by a retrospective analysis of the first consecutive 500 procedures in 443 patients. The indication for thoracoscopic laser pneumoplasty was intractable dyspnea. METHODS: Advanced age (mean age, 67 years), high oxygen dependency (70%), steroid use (46%), and markedly diminished physical capacity (2% bedridden and 27% wheelchair-bound) were noted. Thoracoscopic laser pneumoplasty was carried out under general anesthesia and one-lung ventilation. Type 3 bullae (381 procedures) were contracted by contact neodymium:yttrium-aluminum garnet laser and type 4 bullae (199 procedures) excised. The operative mortality rate was 4.8%. RESULTS: Subjective improvement was reported by 87% of the patients. Follow-up functional evaluation was available in 229 patients, which showed highly significant improvement. A comparison of preoperative and postoperative functional tests between type 3 and 4 bullae patients showed no significant difference, except the latter had higher decrease in airway resistance, residual volume, and total lung capacity. CONCLUSIONS: Thoracoscopic laser pneumoplasty is an effective treatment for both type 3 and 4 bullous emphysema with an acceptable risk.

[Successful treatment of a giant bulla by thoracoscopic excision and laser ablation].

A 39-year-old male complaining of shortness of breath on mild exertion. Radiographs revealed that a giant bulla occupied more than half the area of the right lung field. Thoracoscopic excision of the giant bulla was performed using some autosutures. After the emphysematous lesion was consolidated by laser ablation, it was sutured using PDS thread. The bulla in the left lung was similarly excised 3 weeks after the first procedure. The FEV1.0% improved from 72% to 89% after excision and laser ablation of a giant bulla and bullae. Thoracoscopic excision and laser ablation of a giant bulla appears to be an effective alternative to conventional thoracotomy.