A cytotoxic T lymphocyte inhibits acquired immunodeficiency syndrome virus replication in peripheral blood lymphocytes.

CD8+ (suppressor/cytotoxic) lymphocytes block replication of HIV-1 or the simian immunodeficiency virus of macaques (SIVmac) in PBL of infected individuals. We now show that these CD8+ lymphocytes undergo clonal expansion in vivo after AIDS virus infection of the individual, suggesting they may be antigen-specific T cells. These CD8+ cells block replication of virus in autologous but not MHC class I-mismatched PBL. The inhibitory lymphocytes express the integrin family molecule 4B4 and the CTL-associated S6F1 epitope of LFA-1. Finally, physical contact is required for the CD8+ lymphocyte-mediated inhibition of AIDS virus replication, since this inhibitory function is blocked by anti-LFA-1 and anti-CD8 mAbs. These studies suggest that the cell that inhibits AIDS virus replication in PBL of infected individuals is a CTL.

Remote postconditioning may attenuate ischaemia-reperfusion injury in the murine hindlimb through adenosine receptor activation.

OBJECTIVE: This study aimed to determine the effect and mechanisms of remote postconditioning (RPC) upon ischaemia-reperfusion injury (IRI) in the ischaemic mouse hindlimb. DESIGN: RPC is the brief application of ischaemia to remote organs immediately before reperfusion of an ischaemic target organ, and it is a novel approach to IRI attenuation. MATERIALS AND METHODS: Right hindlimb ischaemia was induced in mice using a rubber tourniquet, the release of which initiated reperfusion. We established RPC by 5 min of ischaemia followed by 5 min of reperfusion in the left hindlimb immediately before right hindlimb reperfusion. The wet/dry ratio of skeletal muscle (degree of tissue oedema), myeloperoxidase (MPO) activity (accumulation of neutrophils), and nitroblue tetrazolium reduction (tissue necrosis) were evaluated. We also intra-peritoneally injected 8-sulphophenyltheophylline (SPT), an adenosine receptor inhibitor, in RPC mice. RESULTS: Wet/dry ratio, MPO activity and tissue necrosis were significantly lower in the RPC group than in the control group, and injection of SPT impaired the protective effect of RPC. CONCLUSIONS: Our results show that RPC attenuated IRI in murine hindlimb ischaemia, possibly through endogenous adenosine receptor activation, and that RPC might serve as a promising therapeutic option for treating serious limb ischaemia.

CD4-Pseudomonas exotoxin conjugates delay but do not fully inhibit human immunodeficiency virus replication in lymphocytes in vitro.

The CD4 molecule is a high affinity receptor for the human immunodeficiency virus (HIV) envelope glycoprotein (gp160 or gp120). This glycoprotein is expressed on the surface membrane of cells infected with HIV. It has, therefore, been suggested that a soluble form of CD4 might be used as a targeting agent to deliver toxins selectively to cells infected with HIV. We demonstrate that CD4-Pseudomonas exotoxin A (PE) conjugates inhibit the proliferation of gp160-transfected Chinese hamster ovary cells and block HIV replication in virus-infected H9 cells. However, this inhibition of HIV replication appears to be incomplete since virus replication occurs following removal of the toxin conjugates from these cultures. Moreover, CD4-PE conjugates delay but do not inhibit HIV replication in human peripheral blood lymphocytes. These studies suggest that such conjugates should be assessed only as potential adjunctive therapies in the acquired immunodeficiency syndrome.

Time dependent impairment of vagally mediated inhibition of noradrenaline release in the dog heart.

The effects of right vagal nerve stimulation on changes in heart rate, mean coronary sinus blood flow, and noradrenaline overflow rate induced by right cardiac sympathetic nerve stimulation were investigated in anaesthetised decentralised, open chest dogs, when autonomic nerve fibres were stimulated tonically at a frequency of 4 or 5 Hz for 2.5 min. Sympathetic nerve stimulation produced a mean(SEM) increase in each of the variables (50(7.0)% in heart rate, 48(6.3)% in coronary sinus blood flow, and 87.0(12.3) ng.min-1 in noradrenaline overflow rate into coronary sinus blood at 30 s), and the increments remained almost constant during continued stimulation. Vagal nerve stimulation induced a decrease of 34(3.4)% in heart rate and of 22(4.5)% in coronary sinus blood flow and a slight reduction (-9(3.2) ng.min-1) in noradrenaline overflow rate at 30 s. Only the decrease in heart rate faded slightly with time. Combined stimulation of the sympathetic and parasympathetic nerves induced a decrease of 19(5.0)% in heart rate and 4(7.5)% in coronary sinus blood flow and an increase of 34.3(12.0) ng.min-1 in noradrenaline overflow rate at 30 s. The decrease in heart rate and coronary sinus blood flow faded, and noradrenaline overflow rate increased significantly to 86.4(17.3) ng.min-1 at 120 s. The temporal changes in heart rate, coronary sinus blood flow, and noradrenaline overflow rate caused by combined nerve stimulation were readily inhibited by treatment with atropine.(ABSTRACT TRUNCATED AT 250 WORDS)

Dynamical relativistic effects in quasielastic 1p-shell proton knockout from 16O

We have measured the cross section for quasielastic 1p-shell proton knockout in the 16O(e,e(')p) reaction at omega = 0.439 GeV and Q2 = 0.8 (GeV/c)(2) for missing momentum P(miss)

A new enzymatic assay for selectively measuring conjugated bilirubin concentration in serum with use of bilirubin oxidase.

A new enzymatic assay for selectively measuring conjugated bilirubin concentration in serum with use of bilirubin oxidase (BOD) has been developed. At pH 5.5 BOD can oxidize only conjugated bilirubin in the presence of reagents such as sodium fluoride and N-acetylcysteine which can decrease BOD reactivity to unconjugated bilirubin and bilirubin covalently bound to albumin (delta bilirubin). The resulting decrease in absorbance at 450 nm is linearly related to the concentration of conjugated bilirubin in serum. The BOD in this new assay was confirmed to oxidize conjugated bilirubin, and neither unconjugated nor delta bilirubin, based on both its reactivity to unconjugated bilirubin and HPLC results. This assay was found to give satisfactory results, such as in terms of the range of measurement, the reproducibility of the results, the lack of interference with coexisting substances in serum and the stability of the reagent solutions, in practical applications. The serum conjugated bilirubin concentrations determined using this assay correlate well with those determined by the HPLC analysis. This assay can be used for accurate monitoring of changes in the conjugated bilirubin concentration in patient sera. These findings suggest that the conjugated bilirubin assay is useful for fractional determination of bilirubin in icteric sera.

Assay of gamma-glutamyltransferase with amino acid dehydrogenases from Bacillus stearothermophilus as auxiliary enzymes.

A spectrophotometric assay for serum gamma-glutamyltransferase (gamma-GT, EC 2.3.2.2) based on the increasing absorbance at 340 nm due to NADH formation is described. Using gamma-glutamyl dipeptides as the donor substrate, amino acids formed by the gamma-glutamyl transfer from the donor substrate to the acceptor substrate glycylglycine are stoichiometrically converted by the corresponding amino acid dehydrogenases from Bacillus stearothermophilus to produce their keto acids and NADH. gamma-Glutamylalanine was found to be the most specific and sensitive donor substrate. The method can be mechanized, is free of interference and correlates well with conventional methods.

Determination of organically-associated trace metals in estuarine sea-water by solvent extraction and atomic-absorption spectrometry.

Chloroform extraction of trace metals (Ni, Cu, Mo, Mn, Cd and Pb) in estuarine sea-water was studied at pH 8 and pH 3, on the basis that the metals would be associated with dissolved organic matter (DOM), which has recently been characterized by reversed-phase liquid chromatography. Ni, Cu, Mo and Mn were extracted more at pH 8 than at pH 3. Cd and Pb were not associated with the DOM at either pH 8 or 3. The percentage of the total dissolved trace metals in sea-water associated with DOM varied from 0 to 14%. The metals extracted into chloroform at pH 8 were assumed to be associated with neutral or weakly basic DOM while at pH 3 they could be associated with either the neutral (or weakly basic) DOM or two types of acidic DOM.

Stabilized enzymatic reagents for measuring glucose, creatine kinase and gamma-glutamyltransferase with thermostable enzymes from a thermophile, Bacillus stearothermophilus.

Stabilized enzymatic reagents for measuring some components in biological fluids have been successfully developed based on two kinds of thermostable enzymes derived from Bacillus stearothermophilus with separation of the reagent into two complementary solutions. The thermostable glucokinase produced was applied to the measurement of glucose and creatine kinase activity, while the alanine dehydrogenase produced was used for the measurement of gamma-glutamyltransferase activity. The enzymatic reagents were also stabilized by developing two separate reagents with an optimum pH for the main reagent components. The stability of the reagents in liquid form was examined at 10 degrees C. It was clearly shown that the reagents for measuring glucose and creatine kinase activity were stable and retained their full capability for accurate measurement in biological fluids for over one year. The alanine dehydrogenase product was stable for at least 40 days.

Interaction and immunological effect of very late antigen-4, 5, and fibronectin in tonsillar lymphocytes and their relation to age.

We examined the expressions of alpha-subunits of very late antigen (VLA)-4 (alpha4) and VLA-5 (alpha5) on tonsillar lymphocytes and the interaction between these integrin receptors and their ligand, fibronectin (FN). Immunohistological and flow cytometric analyses showed that alpha4 and alpha5 were expressed in the lymphoid follicle and were positive on about 10% each of T cells and on 55% and 35% of B cells. When tonsillar B cells were separated by a discontinuous Percoll gradient, the number of alpha4- and 5-positive cells decreased as the cell density went down, while the number of activated cells went up. After in vitro activation of tonsillar B cells by Staphylococcus aureus Cowan I strain (SAC), the expression of alpha5 and the adhesiveness to FN decreased. The increased proliferation of B cells was observed when tonsillar B cells were cultured with immobilized FN. The expressions of alpha-subunits of VLA-4 and VLA-5 on tonsillar T and B lymphocytes increased with age. These results suggest that: i) B cell activation may cause decreased expressions of VLA-4 and -5, which gives a costimulatory effect on B cell activation itself again in cooperation with FN, ii) Increased expressions of VLA-4 and -5 on tonsillar lymphocytes with age may be related to regional immune response of the palatine tonsils.

Expressions of very late antigen-6 and vitronectin receptor, and their interactions to laminin and vitronectin during tonsillar B-cell activation.

This study examined the expressions of a-subunits of very late antigen-6 (VLA-6; alpha 6) and vitronectin receptor (VNR; alpha V) on tonsillar B cells and interactions between those integrins and their respective ligands, laminin (LM) and vitronectin (VN). alpha 6 and alpha V were expressed on about 30 to 40% of tonsillar B cells. When purified tonsillar B cells were separated by a discontinuous Percoll gradient, the number of alpha 6- and alpha V-positive cells decreased as the cell density went down, while the number of activated cells went up. After in vitro activation of tonsillar B cells by Staphylococcus aureus Cowan I strain (SAC), the expressions of alpha 6 and alpha V and their adhesiveness to LM or VN decreased significantly. Increased proliferation of B cells was observed when tonsillar B cells were cultured with immobilized LM or VN. The results of immunohistological staining showed VLA-6, VNR, LM and VN in the follicular area. These results suggest that the expressions of VLA-6 and VNR on tonsillar B cells may be decreased during B cell activation, and the interaction between VLA-6, VNR, and LM, VN may give a costimulatory effect on B cell activation in the follicular area of the tonsil.

Columnar-cell carcinoma of the thyroid.

A case of columnar-cell carcinoma of the thyroid gland occurring in a 77-year-old female is described. Tracheal deviation and calcified anterior neck lesion were showed in X-ray examination. A computed tomography (CT) scan revealed a soft tissue density mass with calcification of the right thyroid lobe and isthmus. A (99m)Tc scintiscan showed a 'cold' nodule, whereas a (201)Tl scintiscan showed a 'hot' in the right thyroid lobe and isthmus. Subtotal thyroidectomy and dissection of paratracheal adipose tissue were performed after fine needle aspiration cytology (FNA) of the thyroid mass had been done. On the FNA smears, there were many papillary fragments, which contained pseudostratified columnar cells. Histopathologically, the tumor had three different types of neoplasm including papillary carcinoma of usual feature, columnar-cell carcinoma and follicular carcinoma. Transitional changes were observed between columnar-cell carcinoma and follicular carcinoma. The patient has been followed up for 3 years, with no evidence of recurrence or metastasis.

Morphometric study of the medial aspect of the human maxillary sinus with special reference to the nasal fontanelle.

OBJECTIVE: The purpose of this study was to clarify configurations of the nasal fontanelle (NF) from the morphometrical point of view, especially variations of its four margins (anterior, posterior, superior, and inferior), for clinical application. METHODS: We used 136 sides of hemi-sectioned heads that were obtained from 119 donated Japanese cadavers (66 men and 53 women with an average age of 77.6+/-12.0). After mucosal examination, the specimens were boiled with a small amount of powdered soap and treated with protease. The residual mucous membrane was then gently removed. These specimens were originally made for our previous study describing variations of the uncinate process (Isobe M, Murakami G, Kataura A. Variations of the uncinate process of the lateral nasal wall with clinical implications, Clin. Anat. 1998;11:295-303). Different series of measurements were conducted based on surgical approaches and angles of observation. RESULTS: The superior margin of the NF is difficult to identify because the ethmoidal infundibulum, which leads the anterior end of the margin upward, often interrupts the superior margin. Because the inferior and posterior margins are modified by thin paper-like bony structures and because the anterior margin is disturbed by variations of the lacrimal bone and/or the inferior turbinate, they are also difficult to identify. Knowing these variables, we evaluated the NF morphometrically. The NF was located 12.6+/-4.3 mm posterior to the anterior nasal spine and 6.6+/-2.2 mm anterior to the sphenopalatine foramen. The size of the NF was 17.9+/-3.2 mm (anteroposterior axis) x 11.5+/-3.0 mm (inferosuperior axis). The lowest orbital floor was located 10 mm below the superior margin of the anterior NF (-10 mm), and the distance was therefore measured as -3.7+/-2.4 mm on average. Viewed from the maxillary sinus, the location of the NF varied along the anteroposterior axis, whereas it was located consistently at the most superior portion of the medial aspect of the sinus wall. CONCLUSION: During endoscopic sinus surgery for tumor resection in the maxillary sinus, a large and primary window should be prepared in the posterior NF, including partial removal of the uncinate process. Preparation of an additional window in the inferior meatus is preferable to enlargement of the primary window. Approaching the anterior NF should be avoided, if possible, due to its complicated configuration as well as its proximity to the orbital floor.

[Clinical significance of creatine kinase isoform assay by a new reagent based on immunoinhibition method].

Clinical significance of creatine kinase (CK) isoform assay by a new diagnostic reagent based on immunoinhibition method was evaluated. In the method, a monoclonal antibody (CKM-G 01), which inhibits MM3 but not MM1 isoform, was used. Serial serum samples from 143 acute myocardial infarction (AMI) patients were examined in hospitals. Isoform ratio (I.R.: defined as follows; inhibited CK activity/uninhibited CK activity) increased most rapidly (whose peak appeared 8.5 h after onset of AMI), compared with total CK (18.3 h) and CK-MB (16.1 h). In 61 patient samples collected within 4 h from onset, I.R. in 25 samples (41%) exceeded reference interval, whereas total CK in only 17 samples (28%) did it. Reference interval of the I.R. was 0.42 +/- 0.33 (mean +/- SD) in 1,246 normal subjects. These results show that the assay of CK isoforms by the reagents based on immunoinhibition method is useful for earlier detection of AMI.

[Tonsillectomy and osteoarthritic disease].

We investigated the clinical outcome of tonsillectomy in 89 cases out of 100 cases of sternocostoclavicular hyperosteosis (SCCH) and 18 cases of rheumatoid arthritis (RA) who went through the operation more than 3 months before in our hospital between April 1984 and May 1993. The effect of tonsillectomy was associated with a decrease in the degree of pain. The symptom improvement was defined as a degree of pain less than 50% of the pre-operative level. The rates of improvement were 80.9% (72/89) in SCCH and 66.7% (12/18) in RA, whole 79% of SCCH patients had pustulosis palmaris et plantaris, a known focal infection of the tonsil. Eight of nine (88.9%) patients and eleven of 13 (84.6%) patients who were positive for the tonsillar provocation test and the annulation test, respectively, showed improved SCCH postoperatively. These results suggest that focal infection of the tonsil may be one of the etiologies of SCCH and RA.

[Efficacy of tonsillectomy for improving skin lesions of Pustulosis palmaris et plantaris--evaluation of 289 cases at the Department of Otolaryngology of Sapporo Medical University].

In recent years, Pustulosis palmaris et plantaris (PPP) has been described as one of the representative second lesions of focal tonsillitis in Japan. We selected 289 cases of PPP, treated with bilateral tonsillectomy at our department over the past 12 years for this clinical evaluation. In 211 of those 289 tonsillectomy cases, follow-up was done for more than 3 months after operation. Prior to tonsillectomy, 196 cases suffered from PPP only, 90 cases had the complication of osteo-arthral lesions such as sterno-costa-clavicular hyperosteosis (SCCH), rheumatoid arthritis, and soon, whereas only 3 cases showed simultaneous lesions of Psoriasis vulgaris. The efficacy of tonsillectomy for improving the skin lesions was classified into 6 grades, mainly based upon individual evaluation by each patient as follows; disappeared, remarkably improved, improved, slightly improved, unchanged, and worsened. According to our 211 cases, post tonsillectomy improvement of skin lesions was very obvious, with disappearance occurring in 54.3% of cases and remarkable improvement in 21.4%. Age of onset and the duration from onset to tonsillectomy showed no association with effectiveness. However, the shorter the duration from onset to tonsillectomy, the fewer days were required for improvement of the skin lesions. The rate of improvement of PPP rose in conjunction with the number of days after tonsillectomy. We conclude that it requires 12 months, at minimum, to determine the efficacy of tonsillectomy as a treatment for PPP.

Ossifying fibroma of the mandible with primary hyperparathyroidism due to non-familial parathyroid adenoma.

Ossifying fibroma is an uncommon benign osteogenic neoplasm arising from cells of the periodontal ligament, typically with a slowly progressive enlargement of the affected bone. The neoplasm sometimes presents with hyperparathyroidism, most of which cases are due to familial parathyroid tumours. We report a rare case of ossifying fibroma of the mandible which showed very rapid growth and presented with primary hyperparathyroidism due to non-familial parathyroid adenoma. Despite improvement of parathyroid dysfunction after removal of the parathyroid adenoma, the tumour continued to grow very aggressively. The case required partial mandibular resection for complete resection of the tumour, and fixation of the remaining mandible with a titanium plate.

Clinical analyses of focus tonsil and related diseases in Japan.

Clinical analyses were performed in groups of patients with several diseases that are regarded as diseases of focus tonsils. These patients underwent tonsillectomy from 1981 to 1993 in our department. They included 289 cases of pustulosis palmaris et plantaris (PPP), 35 cases of psoriasis vulgaris (PV), 100 cases of sternocostoclavicular hyperosteosis (SCCH), and 18 cases of rheumatoid arthritis (RA). Tonsillectomy was found to significantly alleviate clinical symptoms of the above diseases as follows: 88.1% of the PPP cases, 48.6% of the PV cases, 80.9% of the SCCH cases, and 66.7% of the RA cases. Age distribution of the patients, relation between the effect of tonsillectomy and age, sex and other factors were also analyzed.