Clinical and histologic evaluation of the prolapsed third eyelid gland in dogs. Eighteen prolapses of the gland of the third eyelid in 17 Beagles were randomly allocated to 3 groups, which included nontreated (group 1, n = 6), excised (group 2, n = 4), and surgically repositioned (group 3, n = 8) glands. A schirmer tear test (STT) was performed on affected and normal (control) eyes for 5 consecutive days on weeks 0 (baseline), 2, 4, 6, 8, 10, 12, 16, 18, 20, and 24. All prolapsed third eyelid glands were excised and examined histologically. Ten female and 7 male Beagles were used in the prospective study. Mean age at prolapse was 35.1 weeks (range, 6 to 89 weeks). Control STT data revealed a population mean of 22.2 +/- 2.1 mm/min. Complications developed in 4 of 6 eyes when the gland was allowed to remain in a prolapsed position. Complications for group-1 eyes were significantly (P < 0.005) greater than those for eyes in groups 2 and 3 (0 of 12). Comparison of affected and control eye baseline data revealed decreased STT values for eyes with prolapsed glands (P < 0.01). Mean differences between affected and control eyes were 2.2, 2.0, and 3.4 mm/min for groups 1, 2, and 3, respectively. A significant (P < 0.001) decrease in lacrimation (0.2 to 3.1 mm/min) in group-2 eyes was detected after removal of the gland. Tear production for affected eyes of nontreated dogs fluctuated above and below that of control eyes prior to excision of the prolapsed gland of the third eyelid; however, with time, affected and control eye STT values were not significantly different.(ABSTRACT TRUNCATED AT 250 WORDS)


Use of nonsteroidal anti-inflammatory drugs in food animal practice. We attempted to determine the extent to which nonsteroidal anti-inflammatory drugs (NSAID) are used in the treatment of food animals, and whether withdrawal times for milk and slaughter are recommended to clients. A survey questionnaire was mailed to a stratified random sample of 2,000 veterinarians whose practices were at least half food animals. A cross-sectional study was used to examine the responses to determine whether differences existed on the basis of a respondent's geographic location, number of years since graduation from veterinary college, and percentage of practice devoted to beef and dairy cattle. The response rate was 71% (1,424/2,000). Of those practitioners responding, 93% (1,325/1,424) reported using NSAID, with approximately 57 (751/1,322), 24 (327/1,322), and 18% (244/1,322) of respondents reporting use more than once a week, once a week, and 1 to 2 times per month, respectively. Dairy practitioners reported more frequent use than did beef practitioners. Use of flunixin meglumine was reported more frequently than the use of aspirin, phenylbutazone, or dipyrone. Approximately 88% (1,146/1,306) of respondents that used NSAID did so in combination with antibiotics. Withdrawal times for milk and meat were made on the basis of guidelines for the antibiotic. When using NSAID alone, recommendations for withdrawal times for milk and meat varied extensively. Overall, practitioners indicated that NSAID were useful and necessary for the treatment of food-producing animals.


Benefits and costs of using an orally absorbed vaccine to control rabies in raccoons. In November 1989, the epizootic of rabies affecting raccoons in the mid-Atlantic states reached New Jersey. An economic evaluation was conducted in 2 counties first affected by the epizootic to estimate the costs of the epizootic and to assess the costs and benefits of orally administering a newly developed recombinant rabies vaccine to prevent further spread of the disease. Data on expenditures associated with prevention of rabies in human beings and domestic animals and laboratory testing of suspect animals were collected and analyzed for 1988 (before the epizootic) and 1990 (first full year of the epizootic). Benefit-cost ratios were calculated and used to evaluate the economic advisability of the vaccine at various vaccination program alternatives. Two indices of capital investment analysis, payback period and net present value, were used to evaluate the economic benefits of the rabies vaccine. Expenditures were estimated to be $1,952,014 in 1990 (primarily for pet animal vaccinations), compared with $768,488 in 1988. Benefit-cost ratios ranged from 2.21 for the most expensive vaccination program alternative to 6.80 for the least expensive alternative. The payback period varied from 0.69 to 2.11 years, and the net present value ranged from $2,105,453 to $4,877,452. The high costs of this epizootic necessitated the reallocation of scarce public health resources to various rabies prevention activities, particularly the vaccination of dogs. This study also demonstrated the usefulness of benefit-cost analysis in developing public health strategies. Although the mass application of this recombinant vaccine was found to be economically beneficial, other qualitative considerations must be used to supplement these findings.


Epizootic of rabies in raccoons in Maryland from 1981 to 1987. The epizootic of rabies in raccoons entered Maryland in 1981 and systematically moved through the state affecting raccoons in all counties except those of the lower Eastern Shore. A precoded surveillance form was provided to all county and city health departments and data were requested for each animal head submitted for testing for the year 1985. The disease persisted and, in 1987, all counties previously reporting rabies in raccoons also had documented cases in other species. The incidence of rabies in raccoons increased in late winter and peaked in March. Most human exposures occurred during daylight hours and in private yards. Agricultural areas were similarly affected by rabid animals. Analysis of vaccination status of animals exposed to rabid animals gave estimations of statewide vaccination rates for dogs (70%) and cats (28%). The danger of rabies in cats was emphasized by the large number of animals exposed by each rabid cat. The spillover of rabies in raccoons to other species persisted in all jurisdiction through 1992, with periodic upsurges of disease at 3- to 4-year intervals as the raccoon population is replenished. Rabies was not diagnosed in any human beings.


Yohimbine/flumazenil antagonism of hemodynamic alterations induced by a combination of midazolam, xylazine, and butorphanol in dogs. Reversal of hemodynamic alterations induced by midazolam maleate (1.0 mg/kg of body weight), xylazine hydrochloride (0.44 mg/kg), and butorphanol tartrate (0.1 mg/kg) with yohimbine (0.1 mg/kg) and flumazenil (0.25 mg/kg) was evaluated in 5 dogs. The dogs were anesthetized with isoflurane for instrumentation. With return to consciousness, baseline values were recorded, and the midazolam/xylazine/butorphanol mixture with glycopyrrolate was administered IV. Hemodynamic data were recorded for 60 minutes, and then a reversal mixture of yohimbine and flumazenil was administered IV. All variables were measured 1 minute from beginning of the reversal injection. Mean arterial pressure, pulmonary arterial pressure, systemic vascular resistance, and right ventricular stroke work index increased significantly (P < 0.05) above baseline at 60 minutes. Cardiac index and central venous pressure significantly decreased below baseline at 60 minutes. After reversal, mean arterial pressure and central venous pressure significantly decreased from baseline, whereas cardiac index, pulmonary arterial pressure, and right ventricular stroke work index increased significantly above baseline. Heart rate, cardiac index, and right ventricular stroke work index increased significantly above the 60-minute value after reversal. Mean arterial pressure and systemic vascular resistance decreased significantly (P < 0.05) below the 60-minute value after reversal. The hemodynamic alterations accompanying midazolam/xylazine/butorphanol sedation-anesthesia may be rapidly reversed with a combination of yohimbine and flumazenil.


Resolution of generalized dermatophytosis without treatment in dogs. Natural resolution of generalized dermatophytosis reportedly is common in large-animal species. In dogs with generalized dermatophytosis, antifungal treatment usually is prescribed, because the prevalence of natural resolution of dermatophytosis in dogs is not known. Five dogs, 6 to 18 months old, were diagnosed as having generalized dermatophytosis. Each dog was treated with an inert substance given orally with food, once daily. Signs of disease resolved in 3 of the dogs after 4 to 8 weeks of treatment with the inert substance. Two dogs did not improve, so treatment with ketoconazole was initiated. Resolution was achieved in these 2 dogs after 6 weeks of treatment with ketoconazole.


Pneumothorax in a dog infected with Dirofilaria immitis. Rupture of a pulmonary abscess secondary to Dirofilaria immitis infection was the cause of acute pneumothorax in a dog. The dog was euthanatized because it did not respond well to treatment for pneumothorax. Postmortem examination revealed a mass in the right middle lung lobe and adult D immitis in the pulmonary artery. Histologic examination of the lungs revealed microfilariae, suppurative inflammation, thrombi, vasculitis, and fibrosis. Clinicians should include D immitis infection in the differential diagnosis of pneumothorax.


Malignant anterior uveal melanoma with diffuse metastasis in a dog. Enucleation was performed in 10-year-old sexually intact female mixed-breed German Shepherd Dog. Histologic examination revealed that the dog had an uveal amelanotic melanoma of the eye. The tumor consisted of anaplastic cells with a high mitotic index, indicating malignancy. On examination 3 months after enucleation, the dog had difficulty breathing and nasal discharge. Radiography revealed pulmonary metastasis. The dog was euthanatized. Necropsy revealed diffuse metastasis involving various organs.


Aneurysmal bone cyst in a six-month-old dog. A 6-month-old female Yorkshire Terrier was examined because of acute left forelimb lameness secondary to a Salter-Harris type IV fracture of the lateral condyle of the humerus. Radiography revealed an eccentric, osteolytic lesion in the distal humeral metaphysis associated with a pathologic fracture. The limb was amputated, and the dog recovered. Microscopic examination revealed an extensive zone of hemorrhage and dilated coalescent spaces, which were filled with blood. Hemosiderin-laden macrophages and multinucleated giant cells were observed throughout the stroma. On the basis of clinical, radiographic, and histologic examinations, a diagnosis of aneurysmal bone cyst was made. Aneurysmal bone cysts generally have been detected in 11- to 13-year-old, medium- to large-sized dogs. They can develop secondary to malignant processes.


Hypertrophic osteopathy in three horses and a pony. Hypertrophic osteopathy was diagnosed in 3 horses and in a pony, ranging in age from 8 to 21 years. There were 2 females, 1 sexually intact male, and 1 gelding. In 3 animals, hypertrophic osteopathy was associated with pulmonary abscesses, bronchogenic squamous cell carcinoma, and ovarian granulosa-cell tumor, respectively, and resulted in death or euthanasia. Duration of the condition ranged from 1 to 4 months. In 1 horse, hypertrophic osteopathy was believed to be secondary to pregnancy, and resolved following uncomplicated delivery of a live foal.


Excision of the distal sesamoid bone for treatment of infection of the digit in a heifer. An 8-month-old Holstein heifer was evaluated for right hind limb lameness of 3 weeks' duration. Diagnoses were osteomyelitis and fracture of the distal sesamoid bone, septic arthritis of the distal interphalangeal joint, and osteomyelitis of the third phalanx. After excision of a section of tissue from the plantar aspect of the digit and partial section of the deep digital flexor tendon, the distal sesamoid bone was excised. The third phalanx was curetted, and the surgical wound was lavaged. Twenty months after surgery, the heifer was fully weightbearing on the affected digit. Surgical exploration of the digit should be considered an alternative to claw amputation in cattle that have severe digital infections.


Severe renal oxalosis in five young Beefmaster calves. Severe renal oxalosis was diagnosed in 4 male and 1 female purebred Beefmaster calves from herds in southeastern and northwestern United States. Clinical signs included weakness, anorexia, lethargy, alopecia, dehydration, and diarrhea. Results of serum biochemical analysis for 2 calves were consistent with end-stage renal disease. Calves died 2 days to 6 weeks after birth. At necropsy, renal calyces were dilated and contained pale yellow granular calculi. Histologically, there was renal interstitial fibrosis, and cortical and medullary tubules were distended with calcium oxalate crystals. Oxalate crystals were also in the tracheal glands of 1 calf. Severe renal oxalosis in young purebred calves, on widely varied diets, with no known exposure to exogenous oxalates is suggestive of an inherited metabolic defect resulting in primary hyperoxaluria.


Concurrent mammary gland hyperplasia and adrenocortical carcinoma in a domestic ferret. Mammary gland hyperplasia associated with adrenocortical carcinoma in a domestic ferret had a histologic appearance similar to that observed in cats. It is important to consider this hyperplastic condition in the differential diagnosis of mammary gland enlargement in the ferret.


Epididymal swelling attributable to generalized lymphosarcoma in a stallion. Aspermia was diagnosed in a 12-year-old Thoroughbred stallion with generalized lymphosarcoma. Invasion of the epididymus by neoplastic cells caused thickening and enlargement of both epididymes. The testes were not affected. The nodular ultrasonographic architecture was similar to that in previously reported cases of infectious epididymitis.


Vacuum phenomenon associated with osteochondrosis of the scapulohumeral joint in dogs: 100 cases (1985-1991). The vacuum phenomenon was visualized in 20 of 100 scapulohumeral joints with osteochondritic lesions in 65 dogs. The phenomenon was associated with the finding of a cartilage flap, lack of joint effusion, and clinical signs of pain and lameness. The vacuum phenomenon was not observed on radiography of 30 clinically normal contralateral joints, and it could not be induced in 36 clinically normal scapulohumeral joints radiographed under stressed extension.


Stress reactions and stress fractures of the proximal palmar aspect of the third metacarpal bone in horses: 58 cases (1980-1990). The case records, radiographs, and nuclear bone scans of 58 horses with stress reactions or stress fractures of the proximal palmar aspect of the third metacarpal bone (MC3) were reviewed. There were 47 Standardbreds, 4 Quarter Horses, 3 Thoroughbreds, 2 Arabians, 1 Oldenburg, and 1 Pony of America. Fifty-six of the horses were racehorses or performance horses. The mean and median ages of affected horses were 4 and 3 years, respectively. Lameness ranged from mild to severe. Physical findings were usually subtle and included signs of pain on deep palpation of the proximal palmar aspect of MC3 and slight effusion of the middle carpal joint in some cases. Lameness was commonly improved by high palmar and palmar metacarpal nerve blocks or anesthesia of the middle carpal joint. Fifty-three horses had higher than normal radiopharmaceutical uptake in the proximal palmar aspect of MC3 in the left or right limb. The other 5 horses had higher than normal radiopharmaceutical uptake bilaterally. Fifty-six of the 63 limbs with abnormal bone scans also had abnormal radiographs. Treatment consisted of a variable period of rest (1 to 6 months). Healing was best assessed by follow-up bone scans. Of the 45 horses for which follow-up information was adequate, 29 (64%) returned to their previous level of performance.


Inactivation of the p53 gene is not required for tumorigenesis of medullary thyroid carcinoma or pheochromocytoma. A polymerase chain reaction (PCR)-mediated RNase protection analysis was performed to detect subtle genetic alterations of p53 in medullary thyroid carcinoma (MTC) and pheochromocytoma. None of the 30 pheochromocytomas showed abnormal RNase protection patterns. Only one of 32 MTCs showed an abnormal pattern, and subsequent DNA sequencing of the PCR product revealed that it had a G to C transversion in codon 49 that resulted in a change from aspartic acid to histidine. However, this was a sporadic MTC with no specific clinicopathological characteristics. On the basis of a previous report that genes on chromosome 17p were not deleted in MTCs and were relatively infrequently deleted in pheochromocytomas, our results suggest that the p53 gene is not involved in tumorigenesis of MTC or pheochromocytoma.


Precise localization of the excision repair gene, ERCC5, to human chromosome 13q32.3-q33.1 by direct R-banding fluorescence in situ hybridization. The genomic DNA fragment encoding the excision repair gene, ERCC5, was mapped by direct R-banding fluorescence in situ hybridization. The signals were localized to human chromosome 13q32.3-q33.1. This result was in agreement with previous reports, and the gene was assigned to a narrower region.


Effect of pravastatin, a potent 3-hydroxy-3-methylglutaryl-coenzyme A reductase inhibitor, on survival of AH130 hepatoma-bearing rats. 3-Hydroxy-3-methylglutaryl-coenzyme A (HMG-CoA) reductase inhibitor is known to have an inhibitory effect on cell growth in addition to a cholesterol-lowering effect. This study examined the effect of pravastatin, a potent inhibitor of HMG-CoA reductase, on the survival of AH130 hepatoma-bearing rats. Pravastatin (1, 2, or 8 mg/kg body weight) was intraperitoneally injected once a day into tumor-bearing rats. The difference in the survival curves was significant between the controls and the rats treated with 8 mg/kg of pravastatin (P < 0.019 by logrank test) but not between the controls and the rats treated with 1 or 2 mg/kg of the inhibitor. The tumor volume was significantly decreased in the rats treated with 8 mg/kg of pravastatin (P < 0.05). These observations showed that intraperitoneal injection of pravastatin could improve the survival of AH130 hepatoma-bearing rats and had an inhibitory effect on the growth of the ascites form tumor.


Telomere elongation frequently observed during tumor metastasis. Changes in the number of telomere repeat arrays have been examined during metastasis of two mouse tumor cell lines. Telomeres were detected as bands and a smear by pulsed-field gel electrophoresis and gel-hybridization using (TTAGGG)4 as a probe. Very long size variants of telomeres were frequently observed in metastatic nodules. This suggests that at least some of the tumor cells have an ability to elongate telomeres. This elongation may compensate for the continuous loss of telomere repeats due to cell divisions, which would eventually lead to cell death.


Oligonucleotide sequences required for natural killer cell activation. Based on the previous finding that certain 30-mer single-stranded oligodeoxyribonucleotides (oligonucleotides) having particular 6-mer palindromic sequences could induce interferon-alpha and -gamma, and enhance natural killer activity, the present study was carried out to clarify the entire relationship between the activity and the sequence of 30-mer oligonucleotides. The results indicated that the activity depended critically on the presence of particular palindromic sequences including the 5'-CG-3' motif(s). The size and the number of palindromes as well as the extra-palindromic sequences also influenced the activity. An oligonucleotide with a 10-mer palindrome and extra-palindromic oligoguanylate sequences showed the strongest activity among the oligonucleotides tested.


A prospective study of atrophic gastritis and stomach cancer risk. The relation of atrophic gastritis, other gastric lesions and lifestyle factors to stomach cancer risk was prospectively studied among 3,914 subjects who underwent gastroscopic examination and responded to a questionnaire survey at the Aichi Cancer Center Hospital. During 4.4 years of follow-up on average, 45 incident cases of stomach cancer were identified at least three months after the initial examination. If the baseline endoscopic findings indicated the presence of atrophic gastritis, the risk of developing stomach cancer was increased 5.73-fold, compared with no indication at the baseline. The risk further increased with advancing degree of atrophy and increasing extension of atrophy on the lesser curvature. These trends in the relative risks were statistically significant (P = 0.027 and P = 0.041, respectively). The risk of developing stomach cancer was statistically significantly increased among subjects with gastric polyps, but not among those with gastric ulcer. Stomach cancer cases tended to consume more cigarettes, alcohol, rice, pickles and salted fish gut/cod roe and less fruits and vegetables and to have more family histories of stomach cancer than noncases, although these differences were not statistically significant. The results of the present study provide additional evidence on the relation between atrophic gastritis and stomach cancer and suggest a need for intensive follow-up of patients with atrophic gastritis and gastric polyps.


Clinicopathologic characteristics and prognosis of breast cancer patients associated with pregnancy and lactation: analysis of case-control study in Japan. Clinicopathologic characteristics and prognosis of breast cancer patients associated with pregnancy and lactation were clarified by means of a case-control study of matched non-pregnant and non-lactating patients with breast cancer. From 18 institutions in Japan, a total of 192 subjects with breast cancer diagnosed during pregnancy (72 cases) and lactation (120 cases) were collected between 1970 and 1988, accounting for 0.76% of all breast cancer patients. The duration of symptoms was longer and tumor size was larger in the study subjects. Although the disease-free interval was longer than that in the control patients, the survival time was shorter. There was no characteristic difference in histologic type. Vascular invasion and lymph node metastasis were found more frequently in the subjects. The positive rates of estrogen receptor and progesterone receptor were lower in the subjects. The 5- and 10-year survival rates of the study patients were 65% and 55%, respectively, and these survivals were significantly lower than those of the control (P < 0.001). The survival rates were poorer in the subjects, in accordance with stage and lymph node metastasis. The results suggest that most of the patients with breast cancer diagnosed during pregnancy and lactation are in a more advanced stage because of a delay in detection and diagnosis, and hence have unfavorable prognosis. Therefore, it is important to diagnose and treat early for improvement of prognosis in patients with breast cancer during pregnancy and lactation.


Comparison of the anatomic distribution of stomach cancer and precancerous gastric lesions. The anatomic distribution of precancerous gastric lesions among 3,400 residents in Linqu, Shandong Province of China, was compared with the anatomic distribution of stomach cancer (SC) among 959 patients in Tokyo, Japan. The incidence of SC is high in both areas, and locations within the stomach of the precancerous and malignant lesions were classified using similar criteria. Chronic atrophic gastritis (CAG) affected 98% of the population in Linqu, with intestinal metaplasia (IM) the most severe diagnosis in 33% and dysplasia (DYS) in 20%. Neither the SC nor precancerous lesions were uniformly distributed in the stomach. Among the DYS 3% were along the greater curvature of the body, 15% along the lesser curvature of the body, 25% in the angulus, 22% along the lesser curvature of the antrum, and 34% elsewhere in the antrum. Among the SC the corresponding percentages were 2, 16, 28, 25 and 29. The similarity to the SC distribution increased gradually from CAG to IM to DYS, providing further evidence for the multistage progression of precancerous gastric lesions.


An intrinsic thymic epithelial abnormality is responsible for the spontaneous development of predominantly lymphocytic thymomas in BUF/Mna rats. The nature of tumorigenesis of predominantly lymphocytic thymoma was examined using an animal model. Rats of the inbred BUF/Mna strain were found spontaneously to develop predominantly lymphocytic thymomas, histologically indistinguishable from their human counterparts, at an incidence of virtually 100%. Thymic rudiments of BUF/Mna rats grafted 17 months previously under the renal capsule of young athymic ACI/NMs-rnu/rnu rats also gave rise to similar lesions. The lymphocytes in the thymomas expressed T-cell antigens (rat Lyt-1 and Lyt-2.3), as in the normal case, and ACI rat specific antigen. When BUF/Mna rats of thymoma age were irradiated with a lethal dose of 12 Gy and then received a single injection of bone marrow cells (8 x 10(7)) from BALB/c-nu/nu mice, thymomas were re-formed three weeks later (in 2 of 5 rats) with the replacement lymphocytes expressing mouse Thy-1.2 antigen. These results indicate that an intrinsic thymic epithelial abnormality is responsible for the development of predominantly lymphocytic thymomas in BUF/Mna rats.


Accelerated appearance of skin tumors in hairless mice by repeated UV irradiation with initial intense exposure and characterization of the tumors. Skin tumors were produced on the back of hairless mice, HOS (HR/De), by exposure to ultraviolet B light (UVB, 290-320 nm) with 4 different protocols. The first tumors appeared earlier (in 10 weeks in group I and 7 weeks in group III) when initial intense exposure was given, followed by repeated lower-level exposures, than when the mice were exposed to the repeated UV only (in 16 weeks both in group II and group IV). All mice developed skin tumors earlier in the groups given the repeated UV exposures three times a week than in the groups given the exposures twice a week. Most of the skin tumors produced by the UVB exposure were histologically malignant, being transplantable to nude mice, and the cultured cells grown from the tumors were capable of producing tumors when injected into nude mice. The accelerated development of skin tumors by initial intense exposure and short intervals of repeated exposure observed in this study may have implications for humans who expose themselves to intense sunbathing and UV tanning (burning) by fluorescent sun lamps.


Lack of allelic preference in amplification and loss of the c-myc oncogene in methylcholanthrene-induced mouse sarcomas. Sarcomas were induced in F1 mice between C57BL/6N and C3H/He strains by subcutaneous injection of methylcholanthrene. The c-myc oncogene was found to be amplified in 16 cases among 43 sarcomas of C57BL/6N x C3H/He mice and 1 case among 5 sarcomas of the reciprocal cross. The origin of the amplified allele was determined by the polymerase chain reaction single strand conformation polymorphism analysis. Among the 17 sarcomas, only one had both of the alleles amplified. The rest of the tumors carried the amplified c-myc allele coming either from C57BL/6N (9 cases) or from C3H/He (8 cases). These results indicate that the c-myc allele is amplified randomly in methylcholanthrene-induced mouse sarcomas irrespective of its origin, such as paternal or maternal allele and C57BL/6N or C3H/He allele. In addition to these changes, the unamplified c-myc oncogene was found to be lost in 12 cases out of the 17 sarcomas with the amplification.


Genomic organization of the human WT1 gene. We have analyzed the genomic structure of the human WT1 gene, one of the recessive oncogenes for Wilms' tumor at chromosome 11p13. By analyses of three cosmids covering the WT1 gene as well as products generated by polymerase chain reaction, cleavage sites for 10 restriction enzymes were mapped in a region of about 80 kb, and the positions of 10 exons were defined. We also mapped two polymorphic sites for TaqI. Our genomic map will be useful to analyze DNA abnormalities sometimes found in the tumors, as well as loss of heterozygosity.


Quercetin, an inhibitor of heat shock protein synthesis, inhibits the acquisition of thermotolerance in a human colon carcinoma cell line. Here, we describe the effects of quercetin on the induction of thermotolerance as examined by colony forming assay in a cell line derived from human colon carcinoma (COLO320 DM). Cells became resistant to heat treatment at 45 degrees C when they were preheated at 42 degrees C for 1.5 h or at 45 degrees C for 10 min. This induction of thermotolerance was almost completely inhibited by continuous treatment with 100 microM quercetin during the first and second heating sessions, and the interval between. This effect of quercetin was demonstrated to be dose-dependent over a concentration range of 50-200 microM. Quercetin did not increase the thermosensitivity of non-tolerant cells. The presence of quercetin during the first conditioning heating was more effective in inhibiting thermotolerance than its presence during the second heating. Quercetin was also found to inhibit the acquisition of thermotolerance induced by sodium arsenite. Cycloheximide, a nonspecific inhibitor of protein synthesis, did not affect the acquisition of thermotolerance by the same cell line. Quercetin specifically inhibits the synthesis of all heat shock proteins so far reported previously, and this leads to inhibition of the induction of thermotolerance. Such inhibition of thermotolerance by quercetin may improve the efficacy of clinical fractionated hyperthermia.


Flow cytometric analyses of the characteristics of tumor cells treated with two platinum compounds: 1,1-cyclobutanedicarboxylato(2-aminomethylpyrrolidine)- platinum(II) and cisplatin. In order to reduce the toxicities of cisplatin (DDP) and/or to improve antitumor efficacy, a large number of new platinum analogues have been synthesized. 1,1-Cyclobutanedicarboxylato(2-aminomethylpyrrolidine)platin um(II) (DWA2114R) is one of them. In this study, we characterized the action mechanism of DWA2114R flow-cytometrically in 3 human lung cancer cell lines by using bromodeoxyuridine (BrdUrd), rhodamine 123 (Rho) and Ki-67 antibody (Ab), and compared the results with those for DDP. We found that the actions of these 2 platinum analogues were characteristically different at the subcellular level. Our observations may be summarized as follows. a) Simultaneous exposure of cells to DDP and BrdUrd resulted in decreases in fluorescence intensity, i.e. in the amount of BrdUrd incorporated into single-stranded DNA. b) DDP appears to be approximately 20-fold more active than DWA2114R in producing cell cycle perturbation. c) In PC-6 small cell carcinoma cells, DDP induced decreases in S phase cells and accumulation of cells in the G2M phase, whereas in PC-10 squamous carcinoma and PC-3 adenocarcinoma cells DDP produced S phase cell accumulation. Weak but similar changes occurred with DWA2114R. d) The high Ki-67 antigen cell population was decreased by treatment with either DDP or DWA2114R, but DDP reduced the low Ki-67 antigen population more than DWA2114R. e) In PC-10 and PC-6 cells, DDP suppressed Rho incorporation into live mitochondria, whereas DWA2114R produced no change in Rho incorporation. PC-3 cells were not affected by either DDP or DWA2114R. It is likely that these differences reflect the biological activities of DDP and DWA2114R.


Vertical transmission of HTLV-1 in HTLV-1 carrier rat. A female F344 rat was injected with 2.4 x 10(6) MT-2 cells intravenously at 3 and 4 weeks old, and was mated with a non-infected male rat at the 17th week after injection, when the dam rat contained HTLV-1 provirus in the peripheral blood mononuclear cells as determined by polymerase chain reaction. HTLV-1 provirus was detected in at least 2 out of 9 offspring. This system should be useful for studies on the routes and prevention of vertical transmission and on elimination of once-transmitted HTLV-1.


Physical ordering of three polymorphic DNA markers spanning the regions containing a tumor suppressor gene of renal cell carcinoma by three-color fluorescent in situ hybridization. Fluorescent in situ hybridization (FISH) is a powerful technique for gene mapping and multi-color FISH allows us to determine directly the order of two or more probes on both metaphase and interphase chromosomes. We report the physical ordering of three DNA markers by three-color FISH using two fluorochrome dyes, fluorescein isothiocyanate (FITC; green) and rhodamine (red). The third color was visualized as a pseudo-color (yellow) generated by optical interference with FITC and rhodamine. Using this system we could rapidly determine the order of three polymorphic DNA markers located on the 3p23-p21.2 bands which span a tumor suppressor gene for renal cell carcinoma, lung carcinoma, and several other types of tumors.


Sensitive detection of p53 gene mutations in esophageal endoscopic biopsy specimens by cell sorting combined with polymerase chain reaction single-strand conformation polymorphism analysis. For the rapid and sensitive detection of p53 gene mutations in esophageal endoscopic biopsy specimens, we combined cell sorting with the polymerase chain reaction and single-strand conformation polymorphism (PCR-SSCP) analysis. Mutations in exons 5-8 of the p53 gene were investigated by PCR-SSCP analysis using 10(3) sorted nuclei obtained from each endoscopic biopsy specimen of 16 patients with esophageal cancer. DNAs extracted from their respective surgical specimens were investigated by a conventional method of PCR-SSCP analysis. Mutations in the biopsy specimens were detected in 6 of the 12 aneuploid tumors but in none of the 4 diploid tumors. After tumor cell enrichment by cell sorting, one mutation in exon 8 became apparent, which could not be detected from the surgical specimen by a conventional method of PCR-SSCP analysis. This method should improve the sensitivity of detecting p53 gene mutations, and provides additional information concerning the DNA ploidy pattern in the tumors.


The effects of sodium chloride, miso or ethanol on development of intestinal metaplasia after X-irradiation of the rat glandular stomach. The influence of sodium chloride (NaCl), miso (Japanese soybean paste) and ethanol on development of intestinal metaplasia was examined. Five-week-old male CD(SD): Crj rats were treated with two 10 Gy doses of X-rays to the gastric region at a 3-day interval (total 20 Gy). After irradiation, the rats received supplementation with NaCl (1% or 10% in diet), miso (10% in diet) or ethanol (10% in drinking water) for 12 months. The number of alkaline phosphatase-positive foci of intestinal metaplasia in rats given 1% NaCl diet (Group 3) after X-rays was significantly elevated as compared to that in rats given X-rays alone (Group 1) (P < 0.01) or X-rays with 10% NaCl (Group 2) (P < 0.01). In the pyloric gland mucosae, the total numbers of metaplastic foci in rats of Group 3 were much higher than in Group 2, or after miso diet (Group 4) or ethanol supplementation (Group 5) (P < 0.01), but no difference was found between Group 2, 4 or 5 and Group 1. Atypical hyperplasia only appeared at incidences of less than 6% in Groups 1-3 and no promoting effect on gastric tumorigenesis was evident in Group 2. The present results thus showed that the occurrence of intestinal metaplasia induced by X-irradiation can be significantly increased by administration of 1% NaCl and decreased by 10% NaCl and ethanol, but this is not associated with any influence on gastric neoplasia.


Modifying effects of fungal and herb metabolites on azoxymethane-induced intestinal carcinogenesis in rats. Modifying effects of a fungal product, flavoglaucin, and four plant-derived chemicals, shikonin, gingerol, oleanolic acid and paeoniflorin, on intestinal carcinogenesis were examined in a rat model using azoxymethane (AOM). A total of 280 male F344 rats, 6 weeks old, were divided into 12 groups. Group 1 (30 rats) was given two subcutaneous injections of 15 mg/kg of AOM at the start of the experiment. Groups 2 (30 rats), 3 (20 rats), 4 (20 rats), 5 (30 rats) and 6 (30 rats) received a test chemical (flavoglaucin, shikonin, gingerol, oleanolic acid or paeoniflorin, respectively) in the diet at a concentration of 0.02% for 3 weeks, during which time AOM was applied, and then kept on basal diet until the end of experiment (one year). Groups 7-11 (each 20 rats) were given a test chemical corresponding to Groups 2-6, respectively. Group 12 (20 rats) served as a control. The incidence and average number of intestinal tumors in Group 2 (47%, 0.57 +/- 0.68) were significantly less than in Group 1 (74%, 1.07 +/- 0.87) (P < 0.05, respectively). Multiplicity of intestinal neoplasms of Group 3 (0.55 +/- 0.60) or 4 (0.47 +/- 0.51) was also significantly smaller than that of Group 1 (P < 0.05 and P < 0.01, respectively). These results suggest that flavoglaucin, shikonin and gingerol might be promising chemopreventive agents for intestinal neoplasia.


Forestomach neoplasm induction in F344/DuCrj rats and B6C3F1 mice exposed to sesamol. Sesamol was administered at a dietary level of 2% to groups of 30 male and female F344/DuCrj rats and B6C3F1 mice for 104 and 96 weeks, respectively. Squamous cell carcinomas in the forestomach were induced in nine of 29 (31%) effective male rats, three of 30 (10%) female rats, eleven of 29 (38%) male mice and five of 30 (17%) female mice treated with sesamol. Papillomas developed in ten of 29 (34%) male rats and fourteen of 30 (47%) female rats, but not in any of the mice. Hyperplasias developed in almost all rats and mice of both sexes. Significant differences from control values were found for all three lesions in rats and for carcinoma and hyperplasia categories in mice. The incidences of other tumors in the 2% sesamol group were comparable with control values. In conclusion, sesamol induces squamous cell carcinomas in the forestomach of rats and mice, males being more susceptible than females.


3,2'-Dimethyl-4-aminobiphenyl-induced gallbladder carcinogenesis and effects of ethinyl estradiol in hamsters. The effects of ethinyl estradiol (EE) on 3,2'-dimethyl-4-aminobiphenyl (DMAB)-induced carcinogenesis were examined in Syrian golden hamsters. DMAB was subcutaneously injected in corn oil at a concentration of 100 mg/kg once a week for 20 weeks and EE was administered in the diet at a dose of 0.75 ppm throughout the experiment. Some animals were killed at week 20 and all surviving ones were killed at week 50. Gallbladder tumors (adenomas and carcinomas) were induced in 6 of 15 hamsters (40%) in the DMAB + EE group and 5 of 14 (36%) in the DMAB alone group in males, and in 6 of 13 (46%) in the DMAB + EE group and 1 of 8 (13%) in the DMAB alone group in females at week 50. A clearer enhancing effect of EE on DMAB gallbladder carcinogenesis was observed for tumor multiplicity (No./animal) for both sexes; from 0.36 to 0.67 in males and from 0.14 to 0.62 in females. Thus, DMAB was demonstrated to be carcinogenic in the gallbladder of hamsters and EE enhanced this DMAB-induced gallbladder tumorigenesis.


p53 gene mutations associated with anaplastic transformation of human thyroid carcinomas. Anaplastic carcinoma of the thyroid gland, which is one of the most aggressive, malignant tumors in humans, is considered to originate from preexisting differentiated thyroid cancer. To define the genetic alterations associated with such progression, we examined nine cases of anaplastic thyroid carcinoma for mutation in exons 4-9 of the p53 tumor suppressor gene. Preliminary screening for mutation by RNase protection analysis demonstrated that two out of nine anaplastic carcinomas contained sequence alterations in the p53 gene. Subsequent DNA sequencing identified the mutated nucleotides in these two cases; one was a nonsense mutation at codon 165, and the other was a single-base deletion at codon 176 resulting in the creation of a stop codon downstream due to frameshift. The fact that no mutations were detected in coexisting foci of papillary carcinomas from the same patients shows that these mutations of the p53 gene occurred after development of papillary carcinomas. These results suggest that p53 gene mutation triggers the progression from differentiated into anaplastic carcinoma in the human thyroid gland.


Absence of activating mutations in the transmembrane domain of the c-erbB-2 protooncogene in human lung cancer. The rat neu gene is known to be activated by a point mutation in its predicted transmembrane domain. Overexpression of the human homologue of neu, the c-erbB-2 gene, in human lung cancer has been reported, and a similar activating point mutation has been suggested. Therefore, we tested for possible aberrations of the c-erbB-2 gene in the region of the transmembrane domain in surgical specimens of human primary lung cancer from 190 patients, and also examined 24 metastases and 26 specimens of noncancerous portions of the lung of the same patients. Single-strand conformation polymorphism analysis of polymerase chain reaction products revealed no point mutations in the target domain in any of these specimens.


Differences in chemotaxis to fibronectin in weakly and highly metastatic tumor cells. We have examined the chemotactic ability of tumor cell lines with different metastatic potential to plasma fibronectin in Transwell chamber assay. Human renal carcinoma cells with highly metastatic potential, SN12 C-2, chemotactically migrated to fibronectin (10 micrograms/ml) about three-fold more strongly than weakly metastatic SN12 C-4 cells. Similarly, murine melanoma B16-BL6 cells (highly metastatic) showed higher motility to soluble fibronectin in comparison with weakly metastatic B16-F1 cells. Anti-VLA-alpha 3 and beta 1 antibodies potently blocked the chemotaxis of both highly and weakly metastatic cells (SN12 C-2 and C-4) to fibronectin. This implies that the migration of both C-2 and C-4 cells to fibronectin is basically mediated by VLA-3 receptor. In contrast, the anti-VLA-alpha 5 antibody and RGDS peptide significantly inhibited the chemotaxis of SN12 C-2 cells to fibronectin, but did not affect weakly metastatic SN12 C-4 cells. These results suggest that the chemotactic ability to fibronectin positively correlates with the metastatic potential in SN12 and B16 cell lines, and that VLA-5 receptor is concerned in the motility of highly metastatic SN12 C-2 cells to soluble fibronectin.


Recombinant human interferon-alpha 2a increases hormone receptor level of a human breast carcinoma xenograft in nude mice and enhances the anti-proliferative activity of tamoxifen. The effect of recombinant human interferon-alpha 2a (rhIFN-alpha 2a) on the hormone receptor level and antitumor activity of tamoxifen (TAM) was investigated in nude mice using ZR-75-1, an estrogen receptor (ER)-positive, and progesterone receptor (PgR)-negative human breast carcinoma xenograft. ER levels (maximum binding sites) of tumors treated with rhIFN-alpha 2a at a dose of 6 x 10(5) U/mouse/day for 1 or 3 wk were not significantly different from the control, whereas those with rhIFN-alpha 2a at a dose of 6 x 10(4) U/mouse/day for 1 or 3 wk were higher than the control (3.9- to 4.4-fold) with a significant difference at P < 0.01. The increase of ER by rhIFN-alpha 2a was investigated using a sucrose density gradient method. The peak was only seen at 8S in both rhIFN-alpha 2a-treated tumor and control ER, and the sedimentation patterns were almost the same, suggesting that both ERs were essentially equivalent. On the other hand, PgR of all the treated groups could be detected, while that of the control group was undetectable. The antitumor effect of the combination treatment of rhIFN-alpha 2a and TAM was compared with those of single treatments. While rhIFN-alpha 2a at a dose of 6 x 10(5) U/mouse/day and TAM did not show a combination effect, rhIFN-alpha 2a at a dose of 6 x 10(4) U/mouse/day and TAM showed a synergistic combination effect, and ER was decreased to the threshold of detection by the combination treatment. These findings indicated that a low dose of rhIFN-alpha 2a increased the ER levels of ER-positive human breast cancer in vivo as well as in vitro and enhanced the anti-proliferative effect of TAM, and the newly synthesized ER was essentially the same as the original ER.


Analysis by step sectioning of early invasive bladder cancer with special reference to G3.pT1 disease. Eighty cases of cystectomized and step-sectioned pT1 transitional cell carcinomas of various grades were analyzed. During the same period, 30 consecutive cases of pT2 carcinomas were also cystectomized and examined for comparison. This is a cross-sectional study of a variety of cases of bladder cancer, designed to delineate the characteristics of G3.pT1 disease. Tumors in this series had a full set of various pathological findings; grades 1 to 3, stages Ta-T1-T2, papillary-papillonodular-nodular tumor configuration, alpha-beta-gamma type of invasion, presence of lymphatic and venous involvement, and presence of associated carcinoma in situ/dysplasia. There is a distinct tendency of stepwise disease progression, such as grade 1-->3, stage T1-->T2, papillary-->nodular configuration, and alpha-->gamma invasion, these factors being mutually related. We noted a similarity between tumor groups containing G3 component, such as G2-3.pT1, G3 > 2.pT1, G3.pT1, G1-3.pT2, and G3.pT2. These tumors are different from groups such as G1-2.pT1, G2.pT1 and G2 > 3.pT1 in terms of tumor configuration and type of invasion. As regards pT1 having G3 components, early cystectomy seems to be the surest treatment. Whenever one adopts a conservative policy in treatment of these tumors, extreme care should be taken to monitor tumor progression.


Detection of locally recurrent colorectal cancer with radiolabeled monoclonal antibody H-15. H-15 (HT-29-15) is an IgG1 mouse monoclonal antibody (mAb) to a cell surface antigen (molecular mass, 200,000 daltons) present on virtually all colorectal cancers and also in normal pancreatic ducts and bile ducts, but not in other normal tissues. The biological distribution and imaging characteristics of iodine-131 (131I)-labeled mAb H-15 were studied in 5 primary colorectal cancer patients and 9 patients with local recurrence of colorectal cancer. H-15 mAb labeled with 0.5-10 mCi of 131I was administered 7 to 8 days before surgery at 4 dose levels, ranging from 0.2 to 6 mg. Selective mAb H-15 localization to tumor tissues was demonstrated in 6 of 12 patients with antigen-positive tumors: in two patients, recurrent tumors were negative to H-15 mAb, although the primary tumors were positive. In six patients with positive radioimaging, tumor:normal tissue ratios ranged from 2.05 to 5.35 and tumor:serum ratios from 1.18 to 2.73. The clarity of images seems to correlate well with the latter ratios. Technetium-99 (99mTc)-albumin blood pool studies in selected cases showed that local recurrence of colorectal cancers was hypovascular, emphasizing the selective localization of mAb H-15 despite poor blood flow distribution in the tumors. The results altogether demonstrated that radioimmunodetection with 131I mAb H-15 is valuable for differentiating recurrent colorectal cancer from granuloma formation after surgery.


Solid-phase anti-CD3 antibody activation and cryopreservation of human tumor-infiltrating lymphocytes derived from epithelial ovarian cancer. The effect of solid-phase anti-CD3 antibody activation and cryopreservation was evaluated on thirteen samples of tumor-infiltrating lymphocytes (TILs) derived from epithelial ovarian cancer. Seven preparations of TILs were cultured with or without solid-phase anti-CD3 antibody in addition to 100 units/ml of recombinant interleukin-2 (rIL-2). The proliferation rate of all of the seven TIL preparations stimulated by anti-CD3 antibody on the fourth or fifth day of culture was 3.4 to 9.8 times greater than that of lymphocytes cultured with rIL-2 alone. Furthermore, in an experiment with five TIL samples activated with anti-CD3 antibody, three of them showed augmented cytotoxic activity against autologous fresh tumor cells. The population of CD3+/CD8+ TILs was increased after 4-5 weeks of cultivation and CD8+ lymphocytes amounted to over 70% in all of seven preparations tested, whereas two of seven preparations not activated by anti-CD3 antibody were CD3+/CD4(+)-dominant. In addition, nine preparations of TILs cultured with rIL-2 were cryopreserved for several weeks; after recovery from cryopreservation, no major change was observed in cell surface markers, in growth rate or in cytotoxic activity. These results suggest that cryopreserved and/or anti-CD3 antibody-activated lymphocytes could conveniently be employed in a clinical trial of adoptive immunotherapy employing TIL.


Culture and characterization of pulmonary microvascular endothelial cells. Surface proteins were compared in endothelial cells (EC) obtained from bovine peripheral lung, pulmonary artery and vein, and dorsal aorta using sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis. Galactose-containing glycoproteins [molecular weight (M(r)) 160-220 and 40 kDa] binding to the Ricinus communis agglutinin (RCA) and peanut agglutinin (PNA) were selectively observed on pulmonary microvessel EC as compared to EC from pulmonary artery, pulmonary vein, and dorsal aorta. The unique RCA- and PNA-binding profiles of EC from the pulmonary artery and microvessels may be important in characterizing EC from different sites in the pulmonary circulation. The pulmonary microvessel EC monolayer was also 15-fold more restrictive to transendothelial flux of [14C]sucrose (M(r) = 342 Da) than the pulmonary artery EC monolayer. In contrast, the microvessel EC were only six- and twofold more restrictive to the flux of larger tracer molecules, ovalbumin (M(r) 43 kDa) and albumin (M(r) = 69 kDa) than pulmonary artery EC. The greater restrictiveness of pulmonary microvessel EC monolayer indicates a major phenotypic difference in the cultured pulmonary microvessel EC barrier function.


Insulinlike growth factor-1 inhibits cell death induced by cycloheximide in MCF-7 cells: a model system for analyzing control of cell death. Prolonged exposure of cells to the potent protein synthesis inhibitor cycloheximide terminates in cell death. In the present study we investigated the effect of insulinlike growth factor-1, insulin, and epidermal growth factor on cell death induced by cycloheximide in the confluent MCF-7 cells, and correlated this effect to the inhibition rate of protein synthesis. Cell death was evaluated by measuring either dead cells by the trypan blue dye exclusion test or by the release of lactic dehydrogenase into the culture medium. After 48 h incubation, cycloheximide (10 to 50 micrograms/ml) was shown to induce cell death in a concentration-dependent manner. Insulinlike growth factor-1, at physiologic concentrations (0.2 to 5 ng/ml), reduced this cell death. Insulin at supraphysiologic concentrations (1 to 10 micrograms/ml) mimicked the effect of insulinlike growth factor-1, whereas epidermal growth factor (10 to 50 ng/ml) had no effect. More than 90% of protein synthesis measured by [3H]leucine incorporation was inhibited by 10 to 50 micrograms/ml cycloheximide. Insulinlike growth factor-1 and insulin at the concentrations that reduced cell death to control level, had no effect on the protein synthesis inhibition rate induced by cycloheximide. These results indicate that inhibition of cell death by insulinlike growth factor-1 does not depend on protein synthesis and may be mediated via a posttranslational modification effect.


Relative promoter activity in human mammary epithelial cells assayed by transient expression. Chimeric DNA expression vectors containing regulatory sequences proximal to the 5' end of coding sequences for mammalian genes provide valuable tools to study gene expression. Genes coding for easily measured products (reporter genes) can be used to study promoter strength and regulation of gene expression after transient expression of promoter-reporter constructs in mammalian cells. To determine the strength of a variety of mammalian and viral promoter-enhancer sequences in primary cultures of human mammary epithelial cells (HMEC), these sequences were fused to the bacterial chloramphenicol acetyltransferase (CAT) gene and transfected into HMEC using strontium phosphate. The long terminal repeat (LTR) of the endogenous murine leukemia virus AKR-623 was the most potent promoter of transient CAT expression in HMEC. A number of commonly available promoter sequences displayed a wide range of activities in these cells. The glucocorticoid responsive LTR promoter from the murine mammary tumor virus modulated expression of CAT and was sensitive to the concentration of dexamethasone in the growth media. In a similar fashion, the regulatory sequences from the murine metallothionein-1 gene retained responsiveness to zinc concentration in the growth media.


Evidence supporting the role of a proteinaceous, loosely bound extracellular molecule in the cell density signaling between tendon cells. Normal cells in culture respond to cell density by altering their proliferation rates and their pattern of protein expression. Primary avian tendon (PAT) cells are a case in point where procollagen production increases approximately 10-fold at high cell density while proliferation almost ceases. In an earlier report focusing on the cell density regulation of procollagen expression, the signaling mechanism communicating the presence of other cells was shown to have the characteristics of a loosely bound component of the cell layer. Extending these studies to the cell density regulation of proliferation, the cell density signal (CDS) was again shown to be altered by medium agitation, stimulating cell division. Agitation, however, was only disruptive to cell signaling when there was a high ratio of medium to cells. When sufficient cells were present, agitation was less effective. Therefore, the CDS controlling procollagen production and the CDS controlling the inhibition of growth seemed to be linked because the signaling mechanism is disrupted in a parallel manner by agitation. However, the proliferative response of PAT cells is more complex in that there is also a positive influence at moderate cell density (> 2 x 10(4) cells/cm2) on the rate of cell division. As a consequence, PAT cells would not proliferate into an area of low cell density, but within the same dish would rapidly fill an area of moderate density. PAT cells were capable of filling a gap between high cell density areas if the gap was less than 2 mm. Medium agitation also affected cells at low cell density in a different manner. It was inhibitory if all the cells were at low cell density but it was stimulatory if the cells at low cell density were in close proximity to cells at high cell density. In addition, medium conditioned by agitation over cells at a high cell density would stimulate cells at low cell density to divide and grow out into low cell density regions. Using the growth-promoting activity of the conditioned medium as an assay, this component of the CDS was shown to have unique characteristics: heat, pH, dithiothreitol (DTT) stable; tris ion and protease sensitive. By gel exclusion chromatography it was larger than 100 kDa. But after DTT treatment its mobility shifted to < 30 kDa while retaining activity.


Participation of the mitochondrial genome in the differentiation of neuroblastoma cells. Using clonal cell lines isolated from murine neuroblastoma C1300, we investigated the mitochondrial changes related to neuronal differentiation and, more generally, the role played by the mitochondrion in this phenomenon. By different approaches (measurement of the mitochondrial mass, immunoquantification of specific mitochondrial proteins, or incorporation of Rhodamine 123), the differentiation of the inducible clone, N1E-115, was found associated with an important increase of the cellular content in mitochondria. This increase could be observed with differentiating N1E-115 cells maintained in suspension, i.e. under conditions where neurite outgrowth is prevented but other early stages of (biochemical) differentiation continue to occur. That these mitochondrial changes are likely to be correlated with these stages of neuronal differentiation, rather than with simple progression to the postmitotic stage, stems from comparative experiments with clone N1A-103, a neuroblastoma cell line variant that becomes postmitotic after induction but fails to differentiate and shows no modification in its cellular content in mitochondria. In accordance with these observations, chloramphenicol prevents differentiation when added together with the inducer. This effect is probably related to the inhibition of mitochondrial translation rather than to modification of the bioenergetic needs because oligomycine, a potent inhibitor of the mitochondrial ATP synthetase, shows no effect on neurogenesis. As a working hypothesis and in keeping with independently published models, we postulate that products resulting from mitochondrial translation could be involved in the organization of the cytoskeleton or of certain membrane components whose rearrangements should be the prerequisite or the correlates to early stages of neuronal differentiation.


Classification of pervasive developmental disorders: some concepts and practical considerations. Classifications have to meet a variety of purposes. Clinical and research needs are different and there is much to be said for separate clinical and research schemes. Care is needed to ensure that classifications provide an appropriate medium for teaching about diagnosis and do not cause difficulties when used as a "passport" to resources. Principles of classification are considered in relation to the need to take course, as well as symptomatology, into account, and with respect to the neuropsychiatric interface. The value of a multiaxial approach is noted. The pros and cons of autism and pervasive developmental disorders (PDD) as an overall descriptive term, of lumping or splitting, and of different choices with respect to PDD subcategories are discussed.


Three diagnostic systems for autism: DSM-III, DSM-III-R, and ICD-10. ICD-10 draft research criteria for childhood autism were applied to a previously published data set comparing DSM-III and DSM-III-R to clinicians' diagnoses of autism. The ICD-10 approach paralleled clinicians' patterns of diagnosis and, to a lesser extent, the DSM-III system. Relative to either clinicians, DSM-III, or ICD-10 the DSM-III-R system overdiagnosed the presence of autism. Implications for research and for future revision of diagnostic criteria are discussed.


Polymorphonuclear leukocytes-mediated lysis of A431 cells induced by IgG1 mouse anti-epidermal growth factor receptor monoclonal antibodies. The ability of different Fc receptors (Fc gamma R) on IgG-mediated cytotoxicity for human epidermoid carcinoma A431 cells bearing large number of epidermal growth factor receptors (EGFRs) was examined by using two isotypes (IgG1 and IgG2a) of murine monoclonal antibodies (MoAbs) against EGFR in the presence of human monocytes and granulocytes. Two MoAbs (225 and LA1) of the IgG1 isotype exhibited effective cytolytic activity for A431 cells with human polymorphonuclear leukocytes (PMN) rather than with human mononuclear cells (MNC). In contrast, two MoAbs (528 and 579) of the IgG2a-isotype lysed the cells less effectively with PMN than with MNC. Anti-Fc gamma R II (CDw32) MoAb 2E1 inhibited the IgG1-mediated cytotoxicity by PMN, and anti-Fc gamma R III (CD16) MoAb 80H3 did not inhibit the IgG2a-mediated cytotoxicity by MNC. Under these conditions, antibody-dependent cell-mediated cytotoxicity by mouse MoAb IgG1 isotypes resulted from antibody binding to the Fc gamma R II (CDw32) of PMN.


Criteria that optimize the potential of murine embryonic stem cells for in vitro and in vivo developmental studies. Cultured mouse embryonic stem (ES) cells are used for both in vitro and in vivo studies. The uncommitted pluripotent cells provide a model system with which to study cellular differentiation and development; they can also be used as vectors to carry specific mutations into the mouse genome by homologous recombination. To ensure successful integration into the germ line, competent totipotent diploid ES cell lines are selected using a cell injection bioassay that is both time consuming and technically demanding. The prolonged in vitro culture of rapidly dividing ES cells can lead to accumulated changes and chromosomal abnormalities that will compromise the biological function and abrogate germ line transmission of chimeric mice carrying novel genetic mutations. Such in vitro conditions will vary between individual laboratories; for example, differences in the serums used for maintenance. Using a number of different criteria we attempt in this paper to define the parameters that we found to be key factors for optimization of the biological potential of established ES cell lines. The successful integration into the germ line is dependant on acquiring or deriving a competent totipotent mouse ES diploid cell line. In this paper parameters and criteria are defined which we found to be key factors for the optimization of the biological potential of established ES cell lines.


Comparison of DSM-III-R and childhood autism rating scale diagnoses of autism. The purpose of this study was to clarify the issue of whether DSM-III-R (American Psychological Association [APA], 1987) over- or underdiagnoses autism by comparing this diagnostic system to a well-established objective measure of diagnosis, the Childhood Autism Rating Scale (CARS). A secondary goal was to determine which of the 16 criteria are the best discriminators of autism. DSM-III-R, CARS, and clinical diagnoses of 138 consecutive admissions to a statewide program for the diagnosis and treatment of autistic and related communication-handicapped individuals (Division TEACCH in North Carolina) were compared. Results indicated a generally high degree of agreement on the diagnosis of autism using the three systems. Within this treatment-oriented program, the CARS and clinical ratings diagnosed a greater number of cases as autistic than did the DSM-III-R criteria, suggesting that DSM-III-R slightly underdiagnosed autism. The criteria that most strongly related to the diagnosis of autism regardless of the system were lack of awareness of others, abnormal social play, an impaired ability to make friends, abnormal nonverbal communication, stereotypic body movements, and restricted range of interests.


Decreased cultured endothelial cell proliferation in high glucose medium is reversed by antioxidants: new insights on the pathophysiological mechanisms of diabetic vascular complications. Exposure to hyperglycemia slows the rate of proliferation of cultured human endothelial cells. Recently, it has been reported that glucose may autoxidize generating free radicals which have been hypothesized to delay cell replication time. To test whether oxidative stress has an effect on delaying cell replication time in hyperglycemic conditions, human endothelial cells cultured from umbilical veins were incubated in 5 or 20 mM glucose, either alone or in the presence of one of three different antioxidants: superoxide dismutase (SOD), catalase and glutathione (GSH). Cells grown in medium with 5 mM glucose, with or without antioxidants, yielded similar population doubling times and cell cycle phase distributions. Significantly lower growth parameters were observed in cells grown in medium with 20 mM glucose, without antioxidants. The presence of the antioxidant reverted them to almost normal growth. These data show that high glucose levels may delay endothelial cells replication time through the generation of free radicals, suggesting a possible pathophysiological linkage between the high levels of glucose and the development of microvascular complications of diabetes, possibly suggesting a new therapeutic approach to prevent such complications.


A review of the DSM-III-R criteria for autistic disorder. The objective of this paper is to review the psychometric properties of the new DSM-III-R criteria for autism. Five data sets were evaluated according to a set of methodological criteria. The results indicate that the DSM-III-R criteria for autistic disorder have, on average, very good sensitivity, but much lower specificity. The implications of this are (a) greater numbers of children diagnosed as autistic; (b) greater numbers of children misdiagnosed as autistic; and (c) greater heterogeneity among samples of autistic children. In essence, the DSM-III-R criteria act more like screening tests than diagnostic criteria. Conceptual and methodologic issues in the evaluation of diagnostic criteria are discussed.


Basic conditions for the drug selection and transient gene expression in the cultured cell line of Bombyx mori. We established basic conditions for transient gene expression and selection of antibiotics in the cultured cell line of silkworm, Bombyx mori, by use of the promoter of the heat shock protein (hsp70) gene of Drosophila melanogaster. The control promoter (hsp70) promoted the expression of chloramphenicol acetyltransferase (CAT) gene ligated at the downstream, dependent on the orientation of the promoter in the silkworm cell. The cell line is able to be supplied for the promoter assay of the silkworm genes. The concentration for the drug selection was determined as 0.75 mg/ml on neomycin analog, G418 (geneticin).


Pervasive developmental disorders: from DSM-III to DSM-III-R. The present paper provides a brief history of the development of the DSM-III-R (American Psychiatric Association [APA], 1987) section on Pervasive Developmental Disorders. It describes the process by which the contents of the text and criteria for Autistic Disorder and Pervasive Developmental Disorder Not Otherwise Specified were decided and gives the reasons for the changes from DSM-III (APA, 1980) categories and criteria. The paper concludes with a short discussion of critical diagnostic issues.


Is Rett syndrome a subtype of pervasive developmental disorders? The author reviews the issue on whether Rett syndrome (RS) is a subtype of pervasive developmental disorders (PDDs). More than 200 articles of RS have been published in the last 10 years. Internal and external validities of RS have been established by several independent studies. There remains the question whether RS presents clinical features that meet the total criteria for PDDs. The available data seem to support the idea of classifying RS as a subtype of PDDs in the DSM-IV.


Diagnostic assessment in a sample of autistic and developmentally impaired adolescents. A sample of 43 autistic and developmentally impaired adolescents were assessed with the Autism Diagnostic Interview (ADI), DSM-III-R criteria, and the clinician's diagnosis. DSM-III-R criteria for autism have low specificity and agree poorly with the other two definitions. Detailed results of the ADI are provided that confirm the usefulness and discriminant validity of this semi-structured diagnostic interview in a sample of very retarded autistic subjects.


The validity of autistic spectrum disorders: a literature review. The objective of this literature review is to assess the validity of autistic spectrum disorders (ASD). Twenty papers were identified that adequately investigated the internal or external validity of various subtypes of ASD. At least three groups can be distinguished from autism on clinical grounds; an Asperger syndrome subtype, and two atypical subtypes characterized by low IQ and high IQ. However, the evidence that these clinical distinctions carry inferences with respect to etiology, clinical course, and treatment is only suggestive. Nevertheless, the specification of several ASD subtypes might promote further research and resolve many of the nosologic issues with respect to the classification of pervasive developmental disorders (PDDs).


Child and adolescent (early onset) schizophrenia: a review in light of DSM-III-R. Early onset schizophrenia (EOS) is defined as that beginning in childhood or adolescence (under 16 or 17). Studies of EOS are infrequent, and comparative adult figures not always available, but tentative conclusions may be drawn. EOS is more common in males; symptomatology is often undifferentiated; frequencies of homotypic family disorder, premorbid schizotypal personality, and neurodevelopmental abnormalities high; outcome poor but only slightly worse than in adults; response to psychotropic drug treatment probably similar though not properly tested; and confusion with psychotic bipolar disorder particularly common. Onset before language is developed presents special diagnostic difficulties. There are a few reports of autistic children developing schizophrenia but this requires replication. Differences from adult schizophrenia are more marked when onset is in childhood than in adolescence but all are quantitative rather than qualitative suggesting that the disorders are the same and that there should be no separate category for children or adolescents.


Childhood disintegrative disorder: issues for DSM-IV. Childhood disintegrative disorder, also known as Heller syndrome or as disintegrative psychosis, is a relatively uncommon condition which has variably been included in official diagnostic systems. Available evidence regarding the validity of this diagnostic concept, particularly with regard to autism, supports inclusion of the category in DSM-IV. Proposed criteria and narrative description for the disorder are presented.


Endoscopic sphincterotomy: preliminary experience at a university hospital. Endoscopic sphincterotomy (ES) was attempted in 38 patients with biliary calculi. There were 21 patients (55.3%) with common bile duct (CBD) stones following cholecystectomy, 14 patients (36.8%) with intact gall bladder and 3 patients with retained CBD stones along with T tube in the early post-operative period. Endoscopic sphincterotomy was possible in all but one patient and duct clearance was attained in 34 (89.4%) patients. Spontaneous clearance of calculi occurred in 31 (81.6%) patients while 3 patients required instrumental extraction. Four patients failed to clear stones and required surgical intervention. Complications occurred in 4 (10.5%) patients--haemorrhage in two, pancreatitis and cholangitis in one each. One patient died of bleeding on the 4th day following ES while hemostasis was achieved in other after two units of blood. Other complications were managed conservatively without any mortality. Endoscopic sphincterotomy appears to be a simple, effective and safe therapeutic modality for the management of biliary calculi.


Fine needle aspiration cytology, sonography and radionuclide scanning in solitary thyroid nodule. One hundred and ninety three consecutive patients with solitary thyroid nodule (STN), with a mean age of 36.0 +/- 12.8 years and male to female ratio of 5.6:1, were studied. Ninety five percent of patients came from iodine deficient regions. Seventy two percent presented for local neck swelling, 12.4% for hyperthyroid state and in 7.7% STN was discovered incidentally. Scintigraphically, 77.7% of nodules were cold, 12.4% hot and 8.5% warm. Sonographic evaluation did not reveal any characteristic echotexture diagnostic of malignancy, but detected clinically nonpalpable accessory nodules in 20.6% of patients. Fine needle aspiration cytology was positive for malignancy in 6.2% of patients. Features of follicular and Hurthle cell neoplasm were seen in 12.9% of aspirates. Eighty eight (45.6%) STN were resected surgically. Histologically, there was one false positive and one false negative aspirate and a case of parathyroid adenoma on aspiration proved to be parathyroid carcinoma on histology.


Ranitidine--rifampicin interaction.

Ranitidine--rifampicin interaction. Newly diagnosed patients of pulmonary tuberculosis (n = 112) were put on a rifampicin-containing drug regimen. Fifty six patients were also given a placebo tablet twice daily while the other fifty-six were given ranitidine 150 mg twice daily. Gastric pH, gastric emptying time, serum rifampicin levels, urinary total and unchanged rifampicin, serum bilirubin and ALT levels were measured serially. Clinical record of adverse symptoms was maintained. Ranitidine increased the basal as well as post-drug gastric pH without altering the gastric emptying time. Concomitant administration of ranitidine and rifampicin did not alter the absorption, metabolism or excretion of the latter but reduced the frequency of gastrointestinal symptoms.


IgA associated glomerulonephritis.

IgA associated glomerulonephritis. Immunohistological analysis of 1146 renal biopsies revealed IgA associated glomerulonephritis (IgAGN) in 83 (7.24%) patients (33 children, 50 adults). Clinical features were unusually severe in a high proportion. Nephrotic syndrome (NS) responding poorly to prednisolone was found in 24%, hypertension (HT) in 39%, and azotemia in 34% of patients. NS was slightly more frequent in children than in adults, but HT and azotemia occurred twice as often in adults as in children. Histologically, extensive glomerular crescents and sclerosis were prominent. In addition, moderate arteriolitis and arteriolosclerosis and marked tubulointerstitial nephropathy were notable features. Thus, a low incidence and marked severity characterized IgAGN in this study.


Adult respiratory distress syndrome following non-thoracic skeletal trauma. Thirty adult patients with severe non-thoracic skeletal trauma were monitored over the first five days for evidence of adult respiratory distress syndrome (ARDS). Three of the patients developed progressive respiratory distress, refractory hypoxaemia and pulmonary infiltrates suggestive of ARDS. Two patients died while one recovered completely. Close monitoring is suggested for early recognition of the disease and institution of therapy.


Human rabies: modes of transmission.

Human rabies: modes of transmission. In contrast to past experience, transmission of rabies by routes other than bite or licking by rabid animals has been identified in recent years. Man to man spread via corneal transplant is a well recognised phenomenon now. Airborne infection in caves as well as in laboratories among workers dealing with rabies virus has been well documented. There is also a potential risk of infection to attendants of rabies patients. Hence, adequate protective measures should be adopted.


Acupuncture in bronchial asthma.

Acupuncture in bronchial asthma. By now, there is adequate clinical experience of treating bronchial asthma with acupuncture. It has a limited role in treating acute attacks since it is a weak bronchodilator, but it has an excellent prophylactic effect in the long run. Controlled trials have shown that acupuncture causes modest improvement in objective parameters, with significant subjective improvement. However, in all these trials there is a large degree of variation in the technique and methodology used. It is expected that with standardization of the technique as per the guidelines given, future trials will be able to quantify the efficacy of acupuncture in bronchial asthma. In addition, investigation of the mechanism by which acupuncture works may lead to better understanding of the pathophysiology of asthma.


Neuropsychiatric manifestations of typhoid fever. Various neuropsychiatric syndromes in typhoid fever were seen in nine patients of multiple drug resistant (both in vitro and in vivo) Salmonella typhi infection of a total of 270 suspected cases in the last two years. All but one patient received oral norfloxacin (400 mg 12 hourly) for two weeks with complete recovery, while the other patient died. Bacteremia and toxemia seem to be the only common factor responsible for neuropsychiatric complications.


Acute lymphoblastic leukaemia presenting as breast mass. A 14 year old girl presented with bilateral symmetrical involvement of the breast with acute lymphoblastic leukaemia (ALL). Treatment with combined therapy of adriamycin, vincristine and prednisolone produced a remission in the leukaemia with complete resolution of the breast mass.


Acute undifferentiated leukaemia in a patient of aplastic anaemia treated with foetal liver infusions. A 54 year old man with aplastic anaemia was treated initially with androgens, corticosteroids and blood transfusions for 24 months. Subsequently, foetal liver infusions were administered thrice, with transient response after the first two infusions and maintained remission after the third. However, 32 months after the first infusion (56 month from initial symptoms), the patient developed acute undifferentiated leukaemia.


Pathophysiology and treatment of spasticity: report of two cases. Two patients who had severe spinal spasticity with painful flexor spasms were treated with oral baclofen with relief of symptoms. The various drugs to treat severe spastic weakness and flexor spasms and their mode of action are briefly discussed.


Intravenous immunoglobulin therapy of immune thrombocytopenia. High dose intravenous gamma globulin has been used in the therapy of immune thrombocytopenia with variable success. Nine non-splenectomized patients with immune thrombocytopenia (8 ITP, 1 Evans syndrome) were treated with IV IgG. One patient with ITP and the case of Evans syndrome had chronic disease, and the others had acute ITP. All patients had been pretreated with steroids with variable response. The patient with chronic ITP and the one with Evans syndrome responded completely to IgG; remission has lasted for 18 months in both. Two patients with acute ITP responded fully to IgG, but one relapsed after 8 months and the other was lost to follow-up after 1 month. Two patients with acute ITP had partial response to IgG, which could be maintained off steroids in one and on a low dose of steroids in the other. Three patients with acute ITP had no response at all to IgG. High dose IV IgG is useful in selected cases of acute as well as chronic ITP, and previous response to steroids increases the likelihood of response. Splenectomy is not a prerequisite for response.


Assessment of impaired left ventricular diastolic function in patients with coronary artery disease, using radionuclide angiography. Left ventricular (LV) diastolic filling at rest was assessed in 76 patients with coronary artery disease (CAD) and 16 healthy subjects using radionuclide angiography. Peak LV filling rate (PFR), expressed in end diastolic volume per second (EDV/sec), was subnormal in CAD patients (1.95 +/- 0.51 as compared to the normal 3.11 +/- 0.36, P < 0.001) and time to PFR (TPFR) was prolonged (171.1 +/- 79 msec versus 106.6 +/- 25 msec normal, P < 0.001). These indices were also abnormal in 60 patients with normal resting LV ejection fraction (PFR 2.17 +/- 0.48 EDV/sec, TPFR 163.9 +/- 68 msec). Abnormal LV filling at rest (PFR EDV/sec or TPFR 160 msec) was found in 88 percent of all patients with CAD, 85 percent of patients with normal resting LV ejection fraction, and 83 percent of patients without Q waves on resting electrocardiogram. Thus, LV diastolic filling, evaluated non invasively by radionuclide angiography, appears to be abnormal in a high percentage of patients with CAD independent of LV systolic function or previous myocardial infarction.


Enalapril in hypertension.

Enalapril in hypertension. Twenty five patients of mild to moderate uncomplicated essential hypertension and five with severe hypertension were treated with long acting converting enzyme inhibitor enalapril for six weeks. Diuretic was added in those patients who did not respond satisfactorily. Twenty one patients of mild to moderate hypertension had their diastolic blood pressure controlled at the end of the study; fifteen with enalapril alone and six with the help of diuretic. Remaining four showed a relative fall but not to level below 150/90 mm Hg. Only one patient with severe hypertension showed fall to normal levels. Four showed a relative fall but not to the normal level even with the addition of a diuretic. Enalapril is an effective anti hypertensive drug in mild to moderate essential hypertension.


Subcutaneous insulin pulse therapy.

Subcutaneous insulin pulse therapy. Subcutaneous Insulin Pulse Therapy (SIPT) consists of administration of small doses of regular insulin hourly or two hourly in the subcutaneous tissue of anterior abdominal wall through a scalp vein needle. Fifteen Non-Insulin Dependent Diabetes Mellitus (NIDDM) subjects, 8 males and 7 females with mean ages 58 +/- 8.7 years and mean duration of diabetes 11.7 +/- 9.1 years and mean BMI 25.2 +/- 5.64 were admitted for elective surgery. Glycemic control was attempted preoperatively with multiple pre-meal doses of Actrapid MC with a single injection of Monotard MC at bed time. The mean fasting plasma glucose in the 15 subjects with this insulin regimen was 321.28 +/- 69.32 mgm% and the insulin requirement per day was 106.87 +/- 35.77 units. The subjects were put on SIPT for 48 to 72 hours. During SIPT the mean fasting plasma glucose dropped to 123.2 +/- 74.11 mgm% and this marked decline in fasting plasma glucose value was statistically significant (P < .05). The insulin requirement during SIPT was 96.42 +/- 31.36 units, similar to the previous regimen (NS). The subjects were switched back to conventional insulin therapy after SIPT during which period the mean fasting plasma glucose was 125.82 +/- 34.50 mgm% and this value was again significantly lower than the pre SIPT fasting plasma glucose value (P < .05). Insulin requirement during conventional insulin therapy after SIPT was reduced to 71 +/- 21.89 units/day. This dose was significantly lower than the insulin dose administered during SIPT (P < .05).(ABSTRACT TRUNCATED AT 250 WORDS)


A comparison of the clinical efficacy of ranitidine and sucralfate in reflux esophagitis. A total of 40 patients with endoscopically proved reflux esophagitis were treated in a randomised study. Treatment was given for six weeks and consisted of daily doses of either 1 gm sucralfate four times a day or one, 150 mg ranitidine tablet twice daily. Twenty patients received each drug. Clinical evaluation was done weekly and endoscopic evaluation after six weeks. Forty percent patients on ranitidine and 50% patients on sucralfate became asymptomatic at six weeks. Fifty percent patients on ranitidine and 60% patients on sucralfate were endoscopically healed. The results did not show any difference in the effect of ranitidine and sucralfate treatment (p > 0.05). Overall tolerance and compliance in the two forms of treatment was good. Though ranitidine has a more convenient dose schedule and is more cost effective, sucralfate needs further clinical trials.


Fine needle aspiration cytology (FNAC) in the evaluation of acute renal allograft rejection. 310 Fine needle aspirations biopsies (FNAB) were performed in sixty-five live related renal donor transplant recipients in a prospective study over 2 years. 82.8% of FNAB were adequate for opinion and there were no complications after the procedure. Methodology used and interpretation of FNACs was as described by Von Willebrand and Hayry. The procedure of FNAC had a sensitivity of 83.3%, specificity of 98.3% and accuracy of 96.1%, and statistically significant (p < .001) correlation was noted between FNAC and Needle biopsy. The increment in lymphoblasts, lymphocytes and score above 3 was suggestive of acute cellular rejection. Increment in monocytes and macrophages above 1% in addition to other cells, was suggestive of acute vascular rejection. FNAC was found to be a safe, simple and easy procedure with high specificity and sensitivity.


"An analysis of epileptic patients nonresponsive to drugs". Fifty-four consecutively referred patients with uncontrolled epilepsy were subjected to Therapeutic Drug Monitoring on an out patient basis. Regular 2 weekly follow up for a minimum period of 2 months was done, after altering the drug dosage and bringing plasma level(s) within therapeutic range. Plasma levels of Phenobarbitone, Phenytoin and Carbamezepine were done by High Pressure Liquid Chromatography. Eventually, 24 patients were controlled and 30 remained uncontrolled. Significant differences between these 2 groups were found, as regards, duration of epilepsy (p < 0.01), associated mental retardation (p < 0.02), initial carbamazepine dosage and plasma levels in patients on carbamazepine montherapy (p < 0.02 and P < 0.01, respectively) and final phenytoin plasma levels in patients on combined therapy with phenobarbitone and phenytoin (p < 0.05). This study emphasizes the importance of early diagnosis and treatment of epilepsy with the help of plasma level monitoring of anti-epileptic drugs.


Glucoregulatory hormones in hepatic cirrhosis. In the present study hormonal responses to 75 gm oral glucose were studied in 34 biopsy proven cases of hepatic cirrhosis and 15 normal subjects. Though fasting blood glucose was similar in both controls and cirrhotics the latter showed higher glucose values throughout the study. The peak of glucose level in cirrhotics was delayed to 60 minutes. Two of 34 (5.8%) cirrhotics showed marginal fasting hyperglycaemia and 44% had (impaired glucose tolerance (IGT). There was no significant difference (P > 0.05) in blood glucose levels in patients with and without varices. The fasting serum insulin was significantly raised in cirrhotics (24.9 +/- 2.2 vs 8.4 +/- 1.2 mu/ml, p > 0.05). Hyperinsulinaemia was significantly marked in cirrhotics with abnormal Oral glucose tolerance test (OGTT) as compared to those who had normal OGTT. The mean fasting serum insulin concentration in patients with and without varices was similar showing thereby that portasystemic shunt in cirrhotics is not the cause for peripheral hyper-insulinaemia. Basal cortisol was similar in cirrhotics and controls though expected fall in cirrhotics like control was absent. Twelve percent cirrhotics had basal human growth hormone (hGH) more than 10 ng/ml. Forty four percent showed paradoxical rise of hGH. hGH has significantly high (p < 0.01) in cirrhotics with abnormal OGTT as compared to those with normal OGTT.


Efficacy of low-dose cytarabine therapy in myelodysplastic syndromes. The efficacy of low-dose cytarabine given as 10 mg sub-cutaneously 12 hourly for 2 weeks and repeated at 2 to 3 weekly intervals was evaluated prospectively in 15 consecutive patients of myelodysplastic syndrome over a 3 year period. Response to therapy was assessed clinically and by haematological parameters including bone marrow examination. No patient achieved complete remission. Three patients responded partially. Four patients transformed into acute myelogenous leukaemia. Overall, the results of therapy were disappointing as all the 15 patients died within 6 months of diagnosis.


Poncet's disease.

Poncet's disease. Tuberculous rheumatism is not generally accepted as a specific disease entity in Great Britain or the U.S.A. We are reporting a adult male whom we believe suffered from this disease, to open the controversy surrounding its existence with a review of the literature.


Bourneville's tuberous sclerosis associated with double outlet right ventricle and infundibular pulmonary stenosis. The cardiac manifestation usually associated with tuberous sclerosis is rhabdomyoma of the heart. We report a rare association with cyanotic congenital heart disease in the form of double outlet right ventricle with infundibular pulmonary stenosis.


W. bancrofti as a causal agent of polymyositis. A case of polymyositis associated with Bancroftian filariasis in an adult male who presented with generalised painful swelling and weakness of muscles is presented. He had elevated muscle enzymes, a myopathic EMG pattern, focal vasculitis on gastrocnemius muscle biopsy and W. bancrofti in the peripheral blood. There was clinical, biochemical and histopathological evidence of resolution of the disorder and total clearance of microfilaremia with diethyl carbamazine (DEC) therapy.


Chloroform ingestion causing toxic hepatitis. Chloroform ingestion poisoning leading to toxic hepatitis is very uncommon. We report one such case in a 16 yrs old patient.


Hemocompatibility of human whole blood on polymers with a phospholipid polar group and its mechanism. The hemocompatibility of a polymer containing a phospholipid polar group, poly(2-methacryloyloxyethyl phosphorylcholine (MPC)-co-n-butyl methacrylate(BMA)), with human whole blood was evaluated. When human whole blood without an anticoagulant was contacted with polymers, the blood cell adhesion and aggregation on the polymer without the MPC moiety was extensive, and considerable fibrin deposition was observed. This phenomenon was suppressed with an increase in the polymer MPC composition. Thus, the MPC moiety in the copolymer plays an important role in the nonthrombogenic behavior of the copolymer. These results were also confirmed by the whole blood coagulation time on the polymer surface which was determined by Lee-White method. The adsorption of phospholipids and proteins from human plasma on poly(MPC-co-BMA) was investigated to clarify the mechanism of the nonthrombogenicity observed with the polymer. The amount of phospholipids was increased; whereas, adsorbed proteins were decreased with an increase in the MPC composition. From these results, we concluded that the phospholipids adsorbed on poly(MPC-co-BMA) play the most important role in the nonthrombogenicity of the MPC copolymer.


In vitro and in vivo studies on bioabsorbable ultra-high-strength poly(L-lactide) rods. Ultra-high-strength poly(L-lactide) (PLLA) rods were fabricated using a drawing technique. Rods with a diameter of 3.2 mm and a draw ratio of 2.5:1 showed initial bending strength and modulus values of 240 MPa and 13 GPa, respectively. The purpose of this study was to investigate the in vitro and in vivo degradation of PLLA rods with a draw ratio of 2.5:1. The greater the rod diameter, the longer the bending strength was maintained in phosphate buffered saline at 37 degrees C. The bending strength retention of rods (diam. 3.2 mm) implanted in the subcutis of rabbits was almost equal to that of rods in the in vitro study, while those rods implanted in the medullary cavity of rabbit femora showed a slightly lower bending strength retention. Molecular weight was reduced to the greatest extent in the medullary cavity, followed by in the subcutis and in vitro. The weight of PLLA rods in the medullary cavity was reduced by 22% at 52 weeks and by 70% at 78 weeks after implantation. Histologically, no inflammatory or foreign body reaction was observed in the medullary cavity for 52 weeks. The drawn PLLA rods maintained a bending strength exceeding that of human cortical bone in the medullary canal for a period of 8 weeks, suggesting that the drawn PLLA rods may be useful in the repair of fractured human bones.


In vitro and in vivo digestion of collagen covalently immobilized onto the silicone surface. In order to study the in vivo digestion of immobilized collagen and gelatin, these proteins labeled with 125I or fluorescein isothiocyanate (FITC) were covalently immobilized onto silicone surfaces, which were grafted with acrylic acid to introduce carboxyl groups, and implanted subcutaneously in rats and mice. When the proteins were labeled with FITC, the amount of proteins immobilized decreased with the increase of the number of FITC molecules conjugated with the protein molecule. In the wet state, FITC conjugated with the proteins was less stable than 125I. Approximately half of the amount of the immobilized proteins was digested in vivo within the first week and until 5 weeks after implantation the proteins were gradually digested. At that time, the amount of the proteins remaining on the silicone surface ranged from 0.6 to 1.0 microgram/cm2, which was several times larger than the amount of an assumed monolayer adsorption of proteins. Even after 15 weeks, the amount of proteins remaining on the silicone was almost the same as after 5 weeks. No significant difference in digestion was observed between collagen and gelatin, regardless of the labeling agent. Because of less stability and easier handling of FITC and higher stability and more difficult handling of 125I, FITC seems more suitable for short-term and 125I for long-term studies.


Tissue reactions induced by modified poly(vinyl alcohol) hydrogels in rabbit cornea. The purpose of this study was to compare the tissue reactions induced by modified poly(vinyl alcohol) (PVA) hydrogels with other materials implanted in the rabbit cornea. PVA hydrogels with and without covalently bonded collagen were compared to ethylene-vinyl alcohol copolymer, and polysulfone discs. These materials were implanted in the rabbit corneal stroma. The resultant tissue reactions were histologically and clinically studied. The PVA hydrogels had high flux rates of glucose and L-lactic acid in vitro. Covalent immobilization of collagen onto their surface was found to render them more biocompatible. However, excessive ulceration and vascularization of the cornea still occurred in vivo, possibly because of extremely little blinking of the rabbits, which might cause desiccation of the cornea.


The fracture toughness of titanium-fiber-reinforced bone cement. Fracture of the poly(methyl methacrylate) bone cement mantle can lead to the loosening and ultimate failure of cemented total joint prostheses. The addition of fibers to the bone cement increases fracture resistance and may reduce, if not eliminate, in vivo fracturing. This study discusses the effect of incorporating titanium (Ti) fibers on fracture toughness. Essential characteristics of the composite bone cement included a homogeneous and uniform fiber distribution, and a minimal increase in apparent viscosity of the polymerizing cement. Ti fiber contents of 1%, 2%, and 5% by volume increased the fracture toughness over non-reinforced bone cement by up to 56%. Bone cements of two different viscosities were used as matrix material, but when reinforced with the same fiber type and content, they showed no difference in fracture toughness. Four different fiber aspect ratios (68, 125, 227, 417) were tested. At 5% fiber content, there was no statistically significant dependence of fracture toughness on fiber aspect ratio. Scanning electron microscopy revealed important toughening mechanisms such as fiber/matrix debonding, local fracture path alteration, and ductile fiber deformation and fracture. Fiber fracture was evidence that the critical fiber length was exceeded. The surfaces of the Ti fibers were rough and irregular, indicating that a high degree of mechanical interlock between matrix and fiber was likely. The energy absorption contribution of plastic deformation and ductile fracture is absent in brittle fibers, like carbon, but is a distinction of the Ti fibers used in this study.


Mechanical deformation of polymer matrix controlled release devices modulates drug release. When magnetic fields were applied to polymer matrices of ethylene-vinyl acetate copolymer embedded with drug and a small magnet, drug release was increased up to 30-fold above baseline levels. It has been hypothesized that the effect of magnetic stimulation on the release of drugs from these matrices is the transduction of the applied magnetic field into a mechanical deformation of the matrix through motion of the magnet within the matrix. This current study provides support for this hypothesis by demonstrating that repeated pulsatile mechanical deformation of matrices can enhance the release of macromolecules from ethylene-vinyl acetate copolymer matrices. Furthermore, similar modulated release kinetics were obtained with mechanically compressed and magnetically stimulated matrices. We also established that modulation was dependent on the ratio of compression area to matrix volume and that modulation was maximized when this ratio was optimized.


Changes in fibrinogen adsorbed to segmented polyurethanes and hydroxyethylmethacrylate-ethylmethacrylate copolymers. Fibrinogen adsorption from blood to biomaterials may regulate platelet adhesion and thrombus formation because of fibrinogen's central role in the coagulation cascade and its ability to bind specifically to the platelet membrane glycoprotein (GP) IIb-IIIa. Adsorption of fibrinogen from blood plasma to many materials exhibits a maximum with respect to plasma dilution and exposure time (the Vroman effect). In this study fibrinogen adsorption to several polymers was examined to ascertain the influence of controlled changes in surface chemistry on the Vroman effect. The materials included hydroxyethylmethacrylate-ethylmethacrylate (HEMA/EMA) copolymers, Biomer, and a series of segmented polyurethanes (PEUs), two of which contained fluorinated chain extenders. Each material exhibited maximal adsorption of fibrinogen at intermediate plasma concentrations. Little effect of soft-segment type or molecular weight was observed and no significant differences in fibrinogen adsorption to the fluorinated PEUs were seen. Changes in the strength of fibrinogen attachment to these materials with time after adsorption were also assessed. Fibrinogen adsorbed for 1 min was displaced more readily by blood plasma than that adsorbed for 1 h, regardless of the material. The more hydrophobic polymers exhibited greater retention of adsorbed fibrinogen. In addition, the fraction of fibrinogen retained by polyethylene depended on the amount of fibrinogen adsorbed to the surface, being greatest when the surface loading was the least. These studies indicate that spreading or transition of adsorbed fibrinogen molecules from a weakly to tightly bound state is a general consequence of protein adsorption to solid surfaces.