Sensitivity of chronically HIV-1 infected HeLa cells to electrical stimulation.

Use of combination anti-retroviral drug regimens including protease inhibitors dramatically decreased morbidity and mortality rates in HIV-1 infected individuals. However, such combination therapies appear to have many side effects, in addition to the emergence of resistant HIV-1 strains. Therefore, in this study we sought to elucidate novel therapeutic principles against HIV-1 infection. We examined the effects of electrical stimulation on both chronically HIV-1(LAI) infected HeLa cells (P6 HeLa/HIV-1(LAI)) and uninfected cells (P6 HeLa). Cells were cultured on an optically transparent electrode and application of potential at 1.0 V vs Ag/AgCl was performed over time periods ranging from 10 min to 60 min. Both P6 HeLa/HIV-1(LAI) and P6 HeLa cells were progressively damaged as the duration of electrical stimulation increased. However, P6 HeLa/HIV-1(LAl) cells were much more influenced by electrical stimulation than P6 HeLa cells. The difference in damage rate was most obvious at 30 min of electrical stimulation, with damaged cells accounting for about 87% and 4% of P6 HeLa/HIV-1(LAI) and P6 HeLa cells, respectively. After the application of potential for 20 min, the proliferation of P6 HeLa/HIV-1(LAI) cells was markedly inhibited, while the P6 HeLa cells proliferated to an extent similar to that of uninfected cells without application of potential. These results indicate that sensitivity to electrical stimulation is much higher in chronically HIV-1 infected cells than in uninfected cells. This could be considered as a useful new approach against HIV-1 infection.

Effect of electrical stimulation on HIV-1-infected HeLa cells cultured on an electrode surface.

Acquired immunodeficiency syndrome (AIDS) is a disease caused by infection with the human immunodeficiency virus (HIV). Although drug therapy for AIDS is available, problems such as side effects associated with drug therapy and the appearance of resistant HIV strains have arisen. Therefore, therapies based on new principles other than drug treatment are required. In the present study, the effect of electrical stimulation on HIV-1(LAI) chronically infected HeLa (P6 HeLa/HIV-1(LAI)) cells cultured on an electrode surface was examined. The results indicated that sensitivity to electrical stimulation was much higher in P6 HeLa/HIV-1(LAI) cells than in uninfected P6 HeLa cells. When electrical stimulation was applied at 1.0 V (vs. Ag/AgCl) for 20 min, the proportion of damage to cell membrane among P6 HeLa/HIV-1(LAI) cells, as evaluated by Trypan blue staining, was approximately 4 times higher than that for uninfected P6 HeLa cells. Furthermore, in comparison with uninfected P6 HeLa cells, the proliferation of P6 HeLa/HIV-1(LAI) cells was significantly suppressed after electrical stimulation. This technique was proven to selectively kill P6 HeLa/HIV-1(LAI) cells, when compared with uninfected control cells.

Humanized antibody to human interleukin-6 receptor inhibits the development of collagen arthritis in cynomolgus monkeys.

In the present study, we demonstrated the anti-arthritic effect of humanized anti-human IL-6 receptor (IL-6R) antibody, MRA, in cynomolgus monkey. MRA can react with monkey IL-6R and block signal transduction of IL-6. Collagen-induced arthritis (CIA) was induced by twice immunizing with bovine type II collagen (CII) emulsified with complete adjuvant. MRA was intravenously injected once a week, from the day of the first collagen immunization, for 13 weeks. The symptoms of arthritis were evaluated using a visual scoring system and radiography. Inflammatory parameters (C-reactive protein (CRP), fibrinogen, and erythrocyte sedimentation rate (ESR) and concentrations of anti-CII antibody, anti-MRA antibody, and MRA were monitored regularly. At the end of the study, histological evaluation was carried out. MRA, at a dose of 10 mg/kg, gave rise to statistically significant suppression. The elevation of serum CRP and fibrinogen levels and ESR were also inhibited. Furthermore, radiographic and histological examination showed that MRA treatment suppressed joint destruction. Our results demonstrate that IL-6 plays an important role in monkey CIA and that MRA may be an attractive agent for the treatment of rheumatoid arthritis.

[Recovered state of iron in female blood donors--after taking an iron supplement].

An iron supplement was given to female blood donors to prevent iron deficiency after blood donation. The levels of items related to red blood cells and serum ferritin (S-Fer) among 17 iron-takers and 31 non-iron-takers were analyzed continuously. In most of the non-iron-takers, the S-Fer levels did not recover within four weeks after the donation of 200ml or 400ml of blood. In the approximately half of the donors, hemoglobin (Hb) levels did not recover either. However, in 59% of those taking iron supplements three times the amount of iron lost by donation, the levels of Hb and S-Fer recovered within two weeks. In the remaining 41%, the Hb levels recovered within four weeks by taking the iron supplement. Four weeks after blood donation, the incidence of iron deficiency anemia among the non-iron-takers increased from 13% to 23% and the incidence of latent iron deficiency among the non-iron-takers increased from 32% to 42%. However, the incidence of iron deficiency anemia among the iron-takers remained 0% and the incidence of latent iron deficiency among the iron-takers decreased from 12% to 6%.

[The states of recovery of blood counts and serum ferritin levels after blood donation].

Complete blood counts and levels of ferritin in serum (S-Fer) in 62 college students were analyzed continuously to clarify the effects of blood donation. In 14 (45%) of 31 females, hemoglobin (Hb) and S-Fer levels showed decrease of more than 5% and 30%, respectively, four weeks after donation of 200ml or 400ml of blood. In 11 (35%) of the males and 6% of the females, Hb levels increased over 5% during the same period. In 61% of the males and 45% of the females, S-Fer levels decreased. In only 19% of the males, the Hb and S-Fer levels were unchanged. Red blood cell (RBC) counts and Hb levels showed almost the same pattern. In 39% of the males and 48% of the females, the white blood cell (WBC) counts decreased more than 10%. In 10% of the males and 23% of the females, WBC counts increased more than 10% after blood donation. The WBC and RBC counts did not always show the same pattern of variation. After blood donation, the incidence of iron deficiency anemia among females increased from 13% to 23% and the incidence of latent iron deficiency among females increased from 10% to 42%.

A chimeric analog of human and salmon calcitonin eliminates antigenicity and reduces gastrointestinal disturbances.

The minimum region in salmon calcitonin (sCT) which induces antigenicity and gastrointestinal disturbances has been identified by examining the cross-reactivity of several sCT fragments and CT analogs with antisera from sCT-treated patients, and by examining inhibition of gastrointestinal motility of these sCT fragments and CT analogs in conscious dogs. Sixteen residues at the N-terminus of sCT comprised the minimum fragment capable of inducing both activities. Human CT (hCT) showed no antigenicity and a four-order weaker inhibition of gastrointestinal motility than sCT. Based on these data, we synthesized the human and salmon chimeric CT, ACT-15, in which the 16 N-terminal residues were those of hCT and the 16 C-terminal residues were those of sCT. ACT-15 had no cross-reactivity with the antisera and had almost the same weak gastrointestinal inhibition effect as hCT in dog and rat models. Nevertheless, it retained a hypocalcemic activity and an analgesic activity comparable to sCT. These results suggest that the amino acid residues in the N-terminal half of CT are responsible for the formation of antibodies and the induction of gastrointestinal disturbances, but may not influence calcium metabolism or analgesia. Clinical studies of ACT-15 will be needed to confirm this hypothesis.

[Effects of long-term low dose radiation--relationship between chromosomal translocation and the cellular oncogenes].

To clarify the late effects of long-term exposure to low doses of radiation, chromosomal aberrations in the lymphocytes of radiological technologists (RT) were analyzed by the trypsin G-banding method. Structural aberrations were identified in 384 (2.5%) of 15,442 cells analyzed from 53 RT as compared to 177 (1.6%) of 11,136 cells from 36 healthy controls. Most of structural aberrations in both groups was translocations, and this frequency was significantly higher in the RT than in the controls. Translocations of chromosomes, such as Nos. 1, 2, 7 and 14 in RT and Nos. 1, 3, 7 and 14 in controls, was observed in over 7% of the cells. 7/14 translocations were the most frequent reciprocal translocations in RT. However, the frequency of 7/14 translocations was not significantly different between the RT and controls. In these cases, most of the break points were localized in band 14q11-12 and 7q32-36. At the chromosomal sites which were related to the sites of ski, abl, myb, mos, myc, N-myc oncogenes, very low incidences of translocation were detected in RT. However, none of the RT demonstrated abnormal clones of cells with identical chromosomal aberrations.

[Posterior chamber lens implantation concurrent with vitrectomy for proliferative diabetic retinopathy].

Combined operation of cataract removal with posterior chamber intraocular lens implantation and pars plana vitrectomy were performed on 25 eyes in 22 patients with cataract concurrent with diabetic retinopathy. In 21 eyes, extracapsular cataract extraction followed by intraocular lens insertion, aiming at in-the-bag fixation, was performed, and in 4 eyes pars plana lensectomy with anterior capsule left and intraocular lens insertion between the iris and anterior capsule was carried out. Mean postoperative follow-up period was 14 months, ranging from 3 to 32 months. Visual acuity on the last examination was 0.5 or better in 2 eyes (8%), 0.1 or better in 12 eyes (48%), and worse than 0.05 in 9 eyes (36%). Major postoperative complications were fibrin reaction (20 eyes, 80%), pupillary deformation (19 eyes, 76%), pupil capture by intraocular lens (3 eyes, 12%), rhegmatogenous retinal detachment (1 eye, 4%), neovascular glaucoma (2 eyes, 8%), and recurrent vitreous hemorrhage (13 eyes, 52%). Intraocular pressure was well controlled in neovascular glaucoma cases. At the last examination ocular fundus was invisible due to vitreous hemorrhage in two eyes.

Effects of long-term radiation exposure on chromosomal aberrations in radiological technologists.

Chromosomal aberrations in the lymphocytes of radiation technologists (RT) were analyzed by the trypsin G-banding method to study the late effects of long-term exposure to low doses of radiation. Structural aberrations were identified in 384 (2.5%) of 15,442 cells analyzed from 53 RT as compared to 177 (1.6%) of 11,136 cells from 36 healthy controls. Stable aberrations were the most frequent in both groups and were either translocations or deletions. Unstable aberrations were mainly acentric fragments in both groups. The frequency of translocations and acentric fragments was significantly higher in the RT than in the controls and was highest in the RT over 50 years. The highest frequency observed in the greater than 50 age group was attributed to the unknown for cumulative dose prior to introduction of film badges. Frequency of chromosomal aberrations correlated with the estimated dose from the film badges and years of experience of each RT based on the equation y = 0.22 + 0.37D + 4.35D2, where y is overall frequency of chromosomal aberrations and D is the estimated radiation dose in Sv.

An attempt at embryo transfer as a means of controlling Bordetella bronchiseptica infection in the rabbit.

Embryo transfer was attempted in order to control disease in rabbits. Embryos were collected by flushing of the oviducts of donor rabbits on Day 2 of gestation, into small tubes containing the medium, transported within the body warmth of the person carrying the tubes and transferred into the oviducts of SPF pseudopregnant recipients. The time between embryo collection and transfer was 7-8 hours. Ten of 56 embryos derived from Bordetella bronchiseptica infected animals developed into newborns. As a result of bacteriological examination of intranasal exudate in six weanlings, no pathogens were detected. We suggest that embryo transfer is an effective and simple alternative to caesarean operation in Bordetella bronchiseptica infected rabbits.

[Clinical study on the serum carcinoembryonic antigen, immunosuppressive acidic protein and C-reactive protein levels in patients with adult T cell leukemia].

Serum carcinoembryonic antigen (CEA), immunosuppressive acidic protein (IAP) and C-reactive protein (CRP) levels were measured in patients with adult T cell leukemia (ATL) in order to clarify their significance in this disease. Mean (+/- SD) serum CEA levels in 11 patients with acute ATL (3.1 +/- 1.3 ng/ml) and 7 patients with smoldering ATL (3.1 +/- 0.5 ng/ml) were significantly higher than in sera of 222 healthy controls (2.4 +/- 1.3 ng/ml). However, the levels in 7 patients with chronic ATL and healthy controls showed no differences. On the other hand, mean (+/- SD) serum IAP levels in patients with acute ATL (928 +/- 395 micrograms/ml), chronic ATL (487 +/- 125 micrograms/ml) and smoldering ATL (429 +/- 90 micrograms/ml) were significantly higher than in sera of healthy controls (359 +/- 103 micrograms/ml). However, the levels in patients with smoldering ATL and healthy controls showed no differences. Serum IAP levels in crisis in chronic and smoldering ATL were similar to those in patients with acute ATL. 85% of ATL patients with IAP levels above 500 micrograms/ml had CRP levels above 1+. Serum CEA, IAP and CRP levels were serially measured in a number of patients. Serum IAP and CRP levels reflected each patient's clinical course more than serum CEA levels. Overall the simultaneous measurements of serum CEA, IAP and CRP levels revealed a potential usefulness for determination of ATL subtype, and serum IAP and CRP levels may provide a way to evaluate the effectiveness of chemotherapy.

[Clinical study of ectopic eruption of permanent incisors and first molars].

UNLABELLED: Using the X-ray films, plastic models and pathological photograph of 4824 patients seen at the Department of Pedodontics of Iwate Medical University, the patterns of ectopic eruptions of permanent incisors and first molars (restricted to these teeth in this study) were classified into 4 groups (I-IV) to investigate the conditions of its incidence, size of the teeth and dental arch, and conditions at the stage of the mixed and permanent dentitions. RESULTS: 1. The total frequency of ectopic eruptions was 5.6%. No difference based on sex was found. 2. By classification, ectopic eruptions of the upper and lower permanent central incisors (Group I) amounted to more than half of the total frequency. The incidence of ectopic eruptions of the upper and lower permanent first molars (Group III) was very low and amounted to only 4.6% of the total frequency. 3. In Groups I and III, the incidence was higher in the maxilla than in the mandible. 4. Based on sex, the incidence was higher in boys in Group III and in girls in group II. 5. Ectopia of the permanent central incisors was frequently bilateral and those of the lower permanent lateral incisors and upper permanent first molars were frequently unilateral. 6. The mesiodistal width of the ectopically erupted permanent teeth tended to be somewhat enlarged, but showed no significant differences from Ono's mean value. 7. In ectopic eruptions at the area of the incisors, the C-C distance was narrowed with an increase in the ectopically erupted permanent teeth. 8. In patients in whom the permanent dentition was formed without occlusal guidance, the basal arch length and dental arch length were large and basal arch width and dental arch width were small. 9. In cases which were followed from the deciduous dentitions to permanent dentitions, the anterior tooth area at the stage of the mixed dentition was frequently crowded. Occlusal guidance was carried out in more than half of these cases.

Assessment of cardiopulmonary baroreflex function in hypertensive and normotensive subjects with or without hypertensive relatives.

1. To investigate whether cardiopulmonary baroreflex control contributes to the pathogenesis and progression of hypertension, we have evaluated the function of the cardiopulmonary baroreflex in 22 patients with essential hypertension and in 17 volunteers with normotension. The normotensive group consisted of 8 subjects with a family history of hypertension and nine with no family history. 2. Forearm vascular resistance (FVR) and central venous pressure (CVP) were measured under control conditions when -10 mmHg lower body negative pressure was applied; the cardiopulmonary slope (CPS = delta FVR/delta CVP) was calculated as an index of the cardiopulmonary baroreflex function. 3. CPS was significantly higher in hypertensives (6.0 +/- 3.93 [s.d.], P less than 0.01) and also tended to be higher in normotensives with a family history of hypertension (3.9 +/- 3.53, P less than 0.05), compared with normotensives without a family history of hypertension (1.7 +/- 0.88). 4. When the hypertensives were divided into two groups, depending on whether CPS was greater or less than 6.0 units, cardiac wall thickness (20 +/- 1.6 mm vs 23 +/- 3.2 mm, P less than 0.05) and the renal vascular resistance (20.9 +/- 6.52 units vs 28.9 +/- 7.32 units, P less than 0.05) were both significantly higher in the Low CPS group. 5. These findings suggest that cardiopulmonary baroreflex function was augmented even in normotensive subjects with hypertensive relatives, as compared with those without hypertensive subjects. Furthermore, cardiopulmonary baroreflex function was augmented in the early stages of hypertension and diminished further with increasing severity.

Clinical study on the serum levels of immunosuppressive acidic protein in patients with adult T-cell leukemia.

Serum levels of immunosuppressive acidic protein (IAP) were measured in patients with adult T-cell leukemia (ATL) in order to clarify its significance in this disease. The mean levels in patients with both acute (854 +/- 404 micrograms/ml) and chronic ATL (439 +/- 103 micrograms/ml) were significantly higher than in sera of healthy controls (367 +/- 104 micrograms/ml). However, mean levels in patients with smoldering ATL, healthy T-cell lymphotropic virus type 1 (HTLV-1) carriers and healthy controls showed no differences. Levels in crisis in chronic and smoldering ATL were similar to those in patients with acute ATL. Serial measurements of serum IAP in a number of patients revealed that the levels reflected each patient's clinical course, suggesting a potential for use in evaluating the effects of chemotherapy.