In vitro evaluation of BAY Y3118, a new full-spectrum fluoroquinolone. BAY Y3118, 1-cyclopropyl-7-(2,8-diazabicyclo[4.3.0]non-8-yl)-6-fluoro-8- chloro-1,4-dihydro-4-oxo-3-quinolinecarboxylic acid hydrochloride, is a new fluoroquinolone with antibacterial activity against an expanded spectrum of species including Staphylococcus aureus, Staphylococcus epidermidis, Enterococcus faecalis, Enterococcus faecium, Streptococcus pneumoniae, Streptococcus pyogenes, and also anaerobes such as Bacteriodes fragilis and Clostridium perfringens. MIC90s for S. aureus, S. epidermidis, E. faecalis, and S. pneumoniae clinical isolates were 0.125, 0.25, 0.125 and 0.25 micrograms/ml, respectively. Against methicillin- and/or quinolone-resistant S. aureus, MIC50 levels of BAY Y3118 were 10- to 100-fold lower than those of tosufloxacin, sparfloxacin, or ciprofloxacin. The potency of BAY Y3118 against all members of the Enterobacteriaceae generally was equal to or 2-fold greater than that of ciprofloxacin or tosufloxacin. BAY Y3118 was also highly active against Haemophilus influenzae, Moraxella catarrhalis, Acinetobacter spp. and Pseudomonas aeruginosa. Increasing inoculum concentrations had a minimal effect on MIC determinations. The drug was determined to be bactericidal based upon reference MBCs and time-kill curves. From the results presented here, it was concluded that this new compound surpasses other known 4-quinolones both in spectrum and activity and that its further evaluation by in vitro, in vivo, and clinical studies seems warranted.


Antimicrobial activity of subinhibitory concentrations of ciprofloxacin against Pseudomonas aeruginosa as determined by the killing curve method and the postantibiotic effect. This investigation used the postantibiotic effect (PAE) and killing curves to examine the antimicrobial activity of subinhibitory (1/8x, 1/4x and 1/2x MIC) and inhibitory (1x MIC) concentrations of ciprofloxacin against mucoid (M) and nonmucoid (NM) urinary isolates of Pseudomonas aeruginosa. Subinhibitory concentrations (1/8x, 1/4x and 1/2x MIC) of ciprofloxacin produced PAEs with no difference between M and NM strains. For NM strains, those with low MICs (< or = 1.0 mg/l) to ciprofloxacin produced significantly longer PAEs than isolates with high MICs (> 1 mg/l). Killing curve studies demonstrated that subinhibitory concentrations of ciprofloxacin produce little effect (1/8x MIC) or stasis (1/4x and 1/2x MIC) of growth for several hours. Only 1x MIC was bactericidal for several strains. At 1/2x and 1x MIC, bacterial inhibition was greater against NM versus M isolates. The M phenotype of P. aeruginosa reduces killing by ciprofloxacin but not the PAE.


Antibacterial activity of citreamicin-alpha (LL-E 19085 alpha) against gram-positive cocci. In vitro antibacterial activity of 429 clinical isolates of gram-positive cocci was tested against citreamicin-alpha (LL-E 19085-alpha) by the agar dilution method. The microorganisms consisted of 313 isolates of staphylococci and 116 strains of streptococci. In vitro activity of citreamicin-alpha was compared with ampicillin, augmentin, cephalothin, erythromycin and vancomycin. MICs of citreamicin-alpha for staphylococci ranged between 0.12-4.0 micrograms/ml and 0.03-0.12 micrograms/ml for Streptococcus pyogenes. Enterococci, however, were relatively more resistant, requiring 2.0 micrograms/ml of this drug to inhibit 64% of the 62 isolates tested. In vitro activity of this antibacterial agent was far superior to that of ampicillin, augmentin, cephalothin and erythromycin, but equal to or slightly inferior to that of vancomycin.


Escherichia coli resistant to ampicillin/sulbactam. Escherichia coli strains resistant to ampicillin/sulbactam from hospitals in 4 different geographic locations were examined with respect to type and amount of beta-lactamase produced. A total of 5 strains was examined from each region. The isoelectric points of all of the involved beta-lactamases were 5.4, corresponding to TEM-1. Km and Vmax values of the beta-lactamases among the clinical isolates resembled those from the control TEM-1 strain. In an 18-hour broth culture the highly resistant isolates produced 3 times more beta-lactamase as compared to the ampicillin/sulbactam-susceptible isolates. However, the highly resistant strains contained approximately the same amount of plasmid DNA (size of > 6,500 bp) as the susceptible isolates. In transformation experiments, both the resistance and the degree of resistance appeared to have been transferable by plasmids. The mechanism for resistance is likely to be a baseline overproduction of TEM-1 beta-lactamase due to either an alteration in the control of gene expression or simply to an increase in the number of copies of the beta-lactamase gene in the plasmids.


Exposure effects of netilmicin on Enterobacter cloacae clinical isolates. The effects of netilmicin exposure on two aminoglycoside (AMG)-resistant and two AMG-susceptible Enterobacter cloacae isolates were studied. It was found that 1 h incubation of bacterial suspensions with netilmicin at 16 mg/l reduces the amounts of lipopolysaccharides present on the bacterial surface. Also, netilmicin pretreatment increased both the susceptibility to normal human serum and the adhesion to human epithelial cells of all four E. cloacae strains examined.


Chlorpromazine: a drug potentially useful for treating mycobacterial infections. Chlorpromazine (CPZ) is one of several phenothiazines known to have antimicrobial properties. It can inhibit mycobacteria, and was reported in the early literature to improve tuberculosis clinically. CPZ was tested here for its ability to inhibit the replication of Mycobacterium tuberculosis and Mycobacterium avium in cultured normal human macrophages, as determined by counts of viable bacteria at 0, 4, and 7 days after bacterial infection of the macrophages. CPZ inhibited the intracellular bacteria at a concentration range of 0.23-3.6 micrograms/ml, and was more effective intracellularly than extracellularly. It was further tested for its ability to cooperate with isoniazid, streptomycin, pyrazinamide, rifampin, rifabutin, penicillin and ethambutol (EMB) against intramacrophage M. tuberculosis and M. avium. CPZ enhanced the effectiveness of most of the drugs tested against intracellular mycobacteria. However, the combination of CPZ and EMB did not result in augmented antimycobacterial activity.


Randomized comparison of ceftriaxone versus ceftriaxone plus amikacin for the empirical treatment of infections in patients with altered host defense: microbiological and clinical evaluation. Two hundred and eighty-four febrile episodes in immunocompromised patients were treated with ceftriaxone alone or in combination with amikacin. In the ceftriaxone-treated group, 60 out of 143 febrile episodes were microbiologically documented, while in the group receiving the combination therapy, there were 32 out of 140 (p = 0.0007). Gram-positive microorganisms were more common than gram-negative ones, accounting for 59 of the 101 isolated bacteria. The ceftriaxone regimen appeared to have a response rate comparable to the combination regimen (73.91 vs. 78.88%). Superinfections occurred under both regimens.


Inhibition of Haemophilus influenzae adherence to buccal epithelial cells by cefuroxime. We studied the effect of subinhibitory concentrations of cefuroxime on the capacity of Haemophilus influenzae to adhere to buccal epithelial cells (BEC). Two encapsulated strains (serotype b) and two nonencapsulated, nontypable strains were studied. All four strains adhered strongly to BEC, with indices (mean number of bacteria adhering to a single BEC) ranging from 19 to 48. Subinhibitory concentrations of cefuroxime (serial dilutions from MIC/2 to MIC/32) were added to cultures in tryptic soy broth and their effect on adherence was tested after 18 h incubation at 37 degrees C. Adherence was diminished by more than 50% by concentrations of cefuroxime ranging from MIC/2 to MIC/8 and varied according to the strain studied. These results show that the adherence of H. influenzae to BEC is inhibited by subinhibitory concentrations of cefuroxime.


Uptake of flurithromycin by human polymorphonuclear phagocytes: partial characterization of the entry mechanism. The ability of flurithromycin and erythromycin to enter human polymorphonuclear phagocytes were studied and compared by a velocity centrifugation gradient technique. Both macrolides were markedly concentrated by human cells and attained cellular to extracellular concentration ratios (C/E) > or = 10. The incorporation was rapid and essentially complete after 60 min incubation. When PMNs were pretreated with formaldehyde, or incubated at low temperatures (4-25 degrees C) or at low pH, the transport ratios of both molecules were reduced. Sodium fluoride and 2,4-dinitrophenol, which decreased erythromycin uptake, did not affect flurithromycin penetration. Perturbation of cell membrane by phorbol myristate acetate, but not by formyl methionyl leucyl peptide, affected C/E ratios of both antibiotics. The addition of amino acids or nucleosides did not influence their transfer into PMNs.


Activity assays of thiadiazine derivatives on Trichomonas vaginalis and amastigote forms of Trypanosoma cruzi. Eight thiadiazine 1,1-dioxide derivatives were evaluated for antitrichomonal and antitrypanosomal activities. In vivo tests were performed on a murine model for trichomoniasis standardized in our laboratory. The capacity of compounds to clear visceral lesions in experimentally infected animals as well as their effects on the mortality time of mice were used as criteria for activity. One of the thiadiazines (compound 7b) showed an efficacy similar to that obtained with the reference drug metronidazole, although higher doses were required. Its toxicity on cell proliferation in tissue culture was moderate. In vitro assays on amastigote forms of Trypanosoma cruzi were carried out using cultures of Vero cells infected with metacyclic trypomastigotes. For one compound (1) trypanocidal activity resembled that of nifurtimox as assessed by microscopic counts of infected and uninfected cells. Unfortunately, this compound showed a high degree of cytotoxicity on Vero cell cultures.


Plasma ammonia in patients with acute leukemia. Plasma ammonia level (PAL) was studied in 43 cases of acute leukemia (AL). PAL was 39.21 +/- 26.2 mumol/L in normal controls and 38.8 +/- 16.6 mumol/L in leukemic patients before chemotherapy. High PAL was found in 40 cases after chemotherapy. Six cases showed clinical manifestations due to severe hyperammonemia, including dizziness, lethargy, confusion, coma and mental changes of various degree, and there was also respiratory alkalosis. After ammonia-trapping therapy, 4 of the 5 patients recovered. The authors believe that high PAL is not uncommon after chemotherapy in leukemic patients. Respiratory alkalosis and unexplained mental and neurologic changes following intensive chemotherapy are useful clues for the diagnosis of hyperammonemia syndrome. Early diagnosis and treatment with ammonia-trapping may improve the rates of remission and survival.


Insular cistern hematoma. A special type of subarachnoid hemorrhage. The clinical manifestations of 20 cases of insular cistern hematoma were characterized by sudden onset violent headache, vomiting, bloody CSF, and subhyaloid hemorrhage (1/3 cases). CT scan showed semilunar high density image centering on the insular cistern with serrate internal edge. The average volume of the hematoma was 20 ml. By means of cerebral angiography, operation and/or autopsy, 7 cases were identified as aneurysm of middle cerebral artery, 6 as aneurysm of posterior communicating artery, and 2 as amyloid angiopathy. In the other 5 cases, the etiology was unknown.


Effect of human growth hormone on the proliferation of human fetal islet cells in vitro. In this study it was shown that 1000 micrograms/L hGH in serum-free medium promoted the attachment, spreading and formation of monolayer of islet cells. The radioactivity test indicated that hGH significantly stimulated the DNA synthesis of islet cells (P < 0.001). Furthermore, after 48 h exposure to hGH (1000 micrograms/L), both the insulin contents and release of islet cells significantly increased (P < 0.001); hGH also enhanced the responsiveness of fetal islet cells to high-concentration glucose plus theophylline stimulation (P < 0.001). However, the effect of hGH in the medium containing 10% newborn calf serum was not so prominent in comparison with that in the serum-free medium. The morphologic assay for mitotic cells showed a combination of 1000 micrograms/L hGH and 10% serum was the most efficient for inducing the mitosis, the mitotic index being 5.95%. We conclude that hGH is an important growth factor for human islet cells.


Pharmacokinetics of clonazepam in developing rats. Time-concentration studies of clonazepam (CZP) were performed in 7-, 14-, and 28-day-old Sprague-Dawley rats. Blood samples and brains were collected at specific intervals following a single subcutaneous (SC) injection of 2.5 mg/kg of CZP. CZP concentrations were measured both in plasma and brain samples by high performance liquid chromatography. Pharmacokinetic parameters were calculated using R-Strip for each age group. The results were compared between the age groups and with those of a similar study in our laboratory using lower dose CZP (1 mg/kg) SC injections in adult Sprague-Dawley rats. Younger rats had slower, delayed and higher peak concentrations, larger areas under the curve (AUC), longer elimination half lives (T1/2), smaller volumes of distribution (Vd) and slower clearances (C1). 28-day-old rats showed the fastest C1, smallest AUC and largest Vd. These data suggest that the pharmacokinetic behavior of CZP is age-dependent. If this is true in human, it is implicated in dosing of neonatal or pediatric patients.


Effects of atrial natriuretic factor on patients with renovascular hypertension. Previous studies have shown the effects of atrial natriuretic factor (ANF) on rats with renovascular hypertension (RVH). In the present study low dose alpha-hANF (0.025 microgram/kg/min) was administered intravenously for 60 minutes to seven RVH patients. Results demonstrated an inhibition of renin-angiotensin-aldosterone system (RAAS), reduction of plasma catecholamine and arginine vasopressin(AVP), diuresis and natriuresis, increase of hematocrit and creatinine clearance, and slight decrease of blood pressure. These results showed that most factors involved in the establishment and maintenance of RVH are affected by ANF infusion.


Apolipoprotein E polymorphism and coronary heart disease. Serum lipoproteins and apolipoproteins were analyzed in 199 patients with acute myocardial infarction (CCU group), and in 211 normal healthy individuals (control group). It was shown that serum lipoprotein abnormalities, especially elevated low density lipoprotein (LDL) levels, are closely related to atherogenesis in relatively young patients and in subjects with severe coronary lesions. The frequency of apo E4 was higher and that of E2 was lower in the CCU group than in the control group. Apo E mutants, E7 and E5, were also frequent in the CCU group. Subjects with an E3/2 phenotype had reduced LDL and increased VLDL levels, and those with an E4/3 phenotype had increased LDL levels in serum. These results suggest that apo E4 is a positive risk factor and apo E2 a negative risk factor for atherosclerosis. The action of apo E isoproteins to increase or decrease serum LDL levels may be one of the mechanisms of atherosclerosis. Furthermore, the effect of apo E isoforms on serum lipoproteins was evident even in childhood. The mutant apo E binding to LDL receptors were investigated to clarify the metabolism of apo E5 and apo E7. The affinity of apo E5 was twice that of the wild form of apo E3. Apo E7, however, had a lower receptor affinity than apo E3. Therefore, the authors postulate that individuals with apo E5 have increased risk of developing hypercholesterolemia and subsequent atherosclerosis. From the binding data of apo E7, the action of apo E7 is considered similar to that of apo E2.


In vitro and in vivo effects of PGE2 on cementless fixation of implant. The effects of prostaglandin E2(PGE2) in vivo and in vitro on cementless fixation of porous coated implant were studied. The results revealed that high levels of PGE2 in the local tissue benefit bone ingrowth in the early stage of implantation. Co-Cr-Mo alloy itself hampers the growth of osteoblasts and decreases the local secretion of PGE2. In the early stage of arthroplasty, the use of PGE2 inhibitors should be forbidden, and proper loading stimulation is preferable. High level PGE2 of local tissue in the later stage indicates bone resorption.


Local changes in aluminium, calcium and phosphorus content of bone caused by alumina implant. Thirty rabbits were divided equally into 5 groups randomly. A hole, 6 mm in diameter and 2 mm deep, was bored on each iliac crest. Two pieces of alumina were implanted into the hole of one side, while the opposite side served as control. These rabbits were killed on 10, 20, 40, 60 and 90 days after operation. Calcium, phosphorus and aluminium contents of iliac bone on both sides were determined by Inductively Coupled Plasma--Atomic Emission Spectrometry. The results showed that the aluminium content of the implanted side in each group was higher than that of the control and difference was significant in 10, 40 and 60 day groups (P < 0.05). This shows that the implant releases aluminium into the bone. Moreover, the calcium and phosphorus contents were significantly lower on the implanted side than on the control side in 10 and 20 day groups (P < 0.05-0.001). Apparently, the aluminium released from the implant in the early stage can interfere with the local calcium and phosphorus metabolism and delay the mineralization of the bone.


New bone formation in the in vivo implantation of bioceramics. A quantitative analysis. Two kinds of synthetic biomaterial, porous tricalcium phosphate (PTCP) and magnetic porous tricalcium phosphate (MPTCP) ceramic granules were implanted in rat femur. In the period of 4 months, the assessment of serial histological sections, scanning electron microphotographs and quantitative analysis of bone formation in the sections showed that both ceramics are biocompatible and degradable in vivo. More new bone formation occurred in the MPTCP group. Endochondral ossification was seen in both groups. The quantitative analysis in this study is reliable, and may be suitable to the similar experimental models.


Carpal instability associated with fracture of the distal radius. Incidence, influencing factors and pathomechanics. 134 fractures of the distal radius in 132 patients are reviewed to determine the incidence and influencing factors of coexisting carpal in stability. By measurement and analysis of the changes in carpal angles and joint spaces, carpal instability was discovered in 41 fractures, an incidence of 30.6%. Six patterns of instability were observed, including dorsal intercalated segmental instability (DISI), scapholunate dissociation, dorsal and palmar translocations, volar intercalated segmental instability (VISI) and ulnar carpal translocation. Accompanying carpal instability was more often seen in elderly patients. To a certain extent, the patterns of instability were related to the type of fracture and palmar tilt angle (PTA) values. DISI was often seen in fractures with PTA smaller than -15 degrees, while scapholunate dissociation was seen in fractures with PTA between -5 degrees and -20 degrees. The possible pathomechanics of the accompanying carpal instability were deduced based on the kinematics and influencing factors indicated in this study.


Extraskeletal chondrosarcoma. Report of 8 cases. Eight cases of surgically and pathologically verified extraskeletal (soft tissue) chondrosarcoma were analyzed with regard to clinical and X-ray features. Their cardinal clinical aspects included presence of a local soft tissue mass and gradual enlargement of the mass accompanied by increasing pain. The X-ray signs were formation of a soft tissue mass, various forms of calcification in the central area of the tumor presence of saucer-like defect on the cortical surface of neighbouring bone in some cases, and bending deformity of the affected bone. The incidence and sites of predilection, the main X-ray findings, radiological diagnosis and differential diagnosis of the tumor were discussed.


Hepatic cavernous hemangioma. CT findings and pathological basis. CT findings in 113 cases of hepatic hemangioma confirmed pathologically including 52 resected specimens and 8 autopsies were studied in correlation with the typical CT scan of 506 cases of the same disease. Mottling enhancement in the surrounding hepatic parenchyma of hemangioma which never appeared in other kinds of tumor was considered as a pathognomonic sign for diagnosis. It was correlated with the presence of dilated vascular spaces or sinusoids communicating with the tumor itself in the surrounding tissue. The thick wall and thin wall types of hemangioma were discussed.


Tumoral calcium pyrophosphate dihydrate deposition disease. Report of a case with a review of the literature. In a 56-year-old female patient with a mass at the proximal end of the left third finger for 22 years, the lesion enlarged obviously during the last 4 years. Initial diagnosis was chondroma with malignant change, and disarticulation of the finger was performed. Pathological diagnosis was "secondary chondrosarcoma". Microscopically, the mass contained a large amount of amorphous basophilic phosphate deposits surrounded by fibrous capsule. The pyrophosphate deposits consisted of birefringent needle shaped or rhomboid crystals which can be identified by polarizing microscope. It can be distinguished as tumoral calcium pyrophosphate dihydrate deposition disease, differing from tophaceous pseudogout, tumoral calcinosis and chondrocalcinosis. Nine cases were collected from the literature and the clinical, radiological and pathological features were discussed.


Delivery by direct intrauterine transfer of gametes. A nurse, 27, primary sterility for 5 years, had normal menstruation, normal size of uterus anterior. HSG: Rt. tube was not visible, Lt. tube slightly enlarged at ampulla. Ovaries were over-stimulated by clomiphene, HMG and hCG.8 ova were taken through vaginal aspiration under B-mode ultrasound scanning. Direct introduction of ova and washed sperms simultaneously into the uterus on Sept. 19, 1991 resulted in successful single pregnancy up to 36(+1) weeks. A 2 250 g normal baby boy was delivered on May 15, 1992 spontaneously. The base and prospect of this manoeuver are discussed with a reference to Craft's procedure.


[Chemosensitivity testing of adenocystic tongue and gingival cancer cell lines]. Chemosensitivity testing of adenocystic, tongue and gingiva cancer cell lines to 14 antitumor drugs using a tetrazolium-based colorimetric assay (MTT assay) was carried out and the values of the relative antitumor activity (RAA) of the drugs were compared. Adriamycin (ADM), methotrexate (MTX) and fluorouracil (5-FU) showed the most potent RAA against the cell lines while Cantharidin (CTD) did not show RAA. The rank orders of other 10 drugs against each cell line differed from each another.


[The effect of founding condition on the microstructure of medium melting alloy]. The effect of founding technology on the properties of medium melting alloy used in dentistry was studied. The porosity and inclusion for the new alloy as well as the cause and prevention of brittleness for the old alloy during remelting were investigated. By using of dehydration borax as purification agent during melting alloy was illustrated.


[Analysis of erroneous diagnosis on temporomandibular joint arthrography]. In the present study, 433 arthrographic cases with temporomandibular joint dysfunction syndrome were included. 30 cases of them underwent operation and 22 cases underwent digital subtraction arthrographic examination after conventional arthrography. It has been found that arthrographic erroneous diagnosis could be made because the artificial manifestations caused by over-dose of contrast medium into the joint cavity, repeated puncture, anterior displacement of the condyle after injecting contrast medium and so on. In addition, the arthrographic manifestations of dis-clock were also analysed.


[Gingival crevicular aspartate aminotransferase levels in periodontitis patients before and after periodontal treatment]. Gingival crevicular fluid samples were collected from 296 teeth from 40 subjects, including 19 rapidly progressive periodontitis (RPP), 8 chronic adult periodontitis (CAP), 7 marginal gingivitis (MG) and 6 healthy subjects (H). The activities of aspartate aminotransferase (AST) in each sample were tested. The results were as follows: (1) The two groups with destructive periodontal disease (RPP and CAP) had greater GCF-AST levels than that from the two non-destructive groups (MG and H). (2) The GCF-AST activities showed significant positive correlations with clinical periodontal parameters, such as probing depth, attachment loss, bleeding index and suppuration. (3) Four weeks after thorough full-mouth root planing, both clinical parameters and GCF-AST levels decreased significantly. The present study suggests that GCF-AST activity might be a sensitive and objective marker for detection of periodontal tissue destruction and inflammation.


[The relationship between family background and craniofacial morphology in class III malocclusions]. Fifty five children with deciduous dentition and 36 adults were selected from 160 Class III patients, 40 children with normal occlusion used as cephalometric control group. T test was operated between the children with normal occlusion, Class III malocclusion with positive family background and with negative family background separately, and distinct analysis was carried out between the two Class III groups. Distribution of family background of skeletal Class III in children and in adults was chi 2 tested. Results show that craniofacial deformities of deciduous Class III malocclusion with positive family background are not more serious than those with negative one, and the rate of developing a skeletal class III malocclusion for patients with positive family background is not higher than those with negative one. It was implied that the value of family information collected by inquiring and cursorily examining the parents is limited for differential diagnosis between skeletal and functional class III malocclusion and it may mislead the prediction of treatment result in orthodontic clinic.


[Repair of the extracranial facial nerve defect by autogenous vein graft: an experimental study]. In this study, we repaired the extracranial facial nerve defect of rabbit using an autogenous vein graft and compared the results with conventional autogenous nerve graft in function and morphology. A 1 cm segment of the extracranial morphology. A 1 cm segment of the extracranial facial nerve was removed in 30 rabbits, the defect was repaired by a segment of vein or by a segment of nerve or without repair. The gross observation, histologic and histochemical examination, electrophysiological study and nerve fascicle graphics analysis were performed at 4, 8, 12, 20 weeks. The results of this study demonstrated that an autogenous vein as a non-nervous tissue graft can repair the extracranial facial nerve defect. 20 weeks after operation, the results with vein graft were superior to nerve graft.


[The effect of gu chi wan on enzyme histochemistry changes in diabetic rats with experimental periodontal disease]. This study observed the effect of Gu Chi Wan on the enzyme histochemistry changes of diabetic rats with experimental periodontal disease. The results indicate that the experimental group has not the decrease of oxidase-reductase and alkaline phosphatase as seen in control group and demonstrate the protective effect of Gu Chi Wan perhaps acts through regulating metabolism of periodontium of rats with experimental periodontal disease, and then improving the immune response of the host.


[A nutrition formula for adults with burn]. The purpose of the present study was to develop a practical formula for calculating the necessary calorie value for Chinese adults with burns of different extents, and to verify the feasibility of the formula in patients with severe burns. 75 burned adults were entered for the study. Resting energy expenditure, some of the hormones and nutritional indices were assessed. On the basis of the observations on energy expenditure, and in accordance with the Chinese dietary tradition, the formula is established as follows: calorie value (kcal/day) = 1000 x body surface area (m2) + 25 x burned surface area (% TBSA); glucose: fat: protein = 55-60:20-30:15-20; calorie:nitrogen = 150-170:1.


[Effect of surface cooling on free skin transplantation: experimental study and preliminary clinical application]. Based on the fact that low temperature can reduce the oxygen consumption and metabolism of detached tissues, for preserving their viability before revascularization, we designed a computer-controlled surface cooling system, which can maintain a planned low temperature in the skin grafts, and the temperature recovers gradually with the revascularization of the grafts. The experiments on rabbits demonstrated that the temperature difference between the epidermic side and recipient-side was 6-8 degrees C when the surface cooling temperature was maintained at 20 degrees C. No significant adverse effects on the process of revascularization were observed. In three out of 4 patients, he flaps with subcutaneous fat reaching 7-9 mm. in thickness, survived.


[Topical use of lysostaphin for Staphylococcus aureus infection of burn wounds in mice]. The effect of lysostaphin on the burn wounds infected with Staphylococcus aureus was studied in mice. The results showed that the mortality and incidence of bacterial isolation in wounds were 17.2% and 8.3%, respectively, in mice treated by lysostaphin, and the figures were significantly lower than that treated by SD-Ag (40.6% and 100%) or base (44.1% and 94.7%). In lysostaphin group the bacterial count of subeschar tissue was 1325 cfu/g, compared with more than 10(9) cfu/g in both SD-Ag and base groups. The results demonstrate that lysostaphin has powerful killing effect on S. aureus, and may be used as atopical antimicrobial to control burn wound infection with S. aureus.


[In vitro measurement of antibacterial activities of topical antibacterial creams]. A new method of in vitro measurement of antibacterial activity of different antimicrobial creams is introduced in this paper. The tested cream in known quantity is uniformed suspended in a solid medium, and bacteria of known quantity are inoculated. The result is expressed in MIC. In this observation, four creams were tested against 3 bacteria. It is the authors impression that the new method is better than the traditional disc method, as it reflects better the actual antibacterial activity of the drug in a cream base. It is helpful in selecting an effective drug or to evaluate the effectiveness of certain drugs in clinical practice.


[Jump flap with pedicle containing superficial epigastric artery for treatment of exposed cranium after electric burn. Report of 9 cases]. Nine patients with denuded cranium after electric burn were treated by using jump flaps with pedicle containing the superficial epigastric artery. The pedicle flap was first transplanted to the burned area on the wrist, and then retransplanted to the cranial defect. It was successful in all cases. The healing time was only one half of that for the traditional method consisting of multiple trephinings through the outer table and then autoskin grafting on the granulations. It also created a better ground for hair grafting later on. Comparing with free flaps or scalp flaps, it was easier to operate and more reliable in blood supply.


[Autodermabrader in the early operation for deep second degree burns. A new operative mode in burns]. Thirty-five cases of deep second burn of the hand (40 hands) were treated with autodermabrader to remove necrotic tissue. In comparison with 30 cases treated with tangential excision, bleeding was less and operation time was decreased markedly (P < 0.01) in the autodermabrader group. Histopathological study of the dermabrased wound was made to evaluate the mechanism of decreased bleeding. We consider that the new operative modality with autodermabader could be used in the treatment of deep II degree burn in exposed areas.


[A new method of expanding free skin grafts]. The harvested skin was processed by a skin-cutting-machine into strips of 0.3 or 0.5 cm in width. Then, the skin strips were glued on alternate folds of a pleated sheet of paper, each fold of which was 0.3 cm or 0.5 cm in width. The paper with skin strips on it was spread flat and cut across into 0.3 cm or 0.5 cm strips. These strips were again glued to another sheet of plicate paper. After being cut across again, many small blocks of skin in equidistance were finally obtained. They were then grafted on the recipient site. The area of grafted wound was four fold or more of the original area of the harvested skin. This method has been used in 6 patients with good result.


[Management of stage III primary breast cancer. An analysis of 221 cases]. From Jan. 1978 to Dec. 1987, 221 patients of stage III breast cancer were treated by surgery combined with adjuvant chemotherapy and/or radiation therapy. The overall 5-year survival rate was 50.4%. The 5-year survival rate in patients with negative lymph node was 72.3% as compared with 37.5% in patients with more than 7 lymph nodes involved (P < 0.05). In patients who received postoperative adjuvant chemotherapy, the 5-year survival rate in premenopausal or postmenopausal group was 62.1% and 41.4% respectively (P < 0.05). The regional lymph node recurrence rate was 4.0% in patients who received postoperative radiotherapy as opposed to 9.6% for those without radiotherapy postoperatively. The distant metastasis rate was 19.1% in lymph node negative group as compared with 45.9% in patients with more than 7 lymph nodes involved (P < 0.05). To decrease the distant metastasis will improve the survival rate in the treatment of breast cancer. We believe that preoperative chemotherapy combined with radical surgery and postoperative adjuvant therapy may improve the survival rate in stage III breast cancer.


[Lymphatic metastasis of breast cancer and selection of operation]. Studies of 169 cases of breast carcinoma showed that lymphatic metastasis was closely related to biological behavior of the cancer. Modified radical operation or modified extended radical operation were performed in cases of TNM stage I and II by pre- and intraoperative estimation of the tumor less than or about 5 cm in diameter, limited within mammary gland, and of expansion type. Histological typing and grading, degree of infiltration, condition of ER and lymphatic metastasis were used to plan postoperative adjuvant therapy. The importance of investigation of ER status of the lesions lies in proper arrangement of combined therapy and prediction of prognosis.


[Infrared light scanning in differentiating benign and malignant breast diseases]. One hundred and fifty patients with various breast diseases were studied using infrared light scanning. The results were compared with those of X-ray mammography and BUS. In 42 cases of breast cancer, 97% were correctly diagnosed using infrared light scanning. The method was significantly superior to X-ray (60%) and BUS (83%). To detect benign diseases, including adenofibromas (30 cases) and hyperplasias (21 cases), infrared light scanning was useful in this series in contrast to X-ray and BUS.


[Doppler color flow imaging for demonstrating changes in portal hemodynamic after Hassab procedure]. Doppler color flow imaging (DCFI) was used to observe portal hemodynamic changes in 40 patients with portal hypertension after Hassab procedure. Preoperative portal blood flow was hepatopetal in all patients, and in 30 of them gastric coronary vein was shown, with the hepatofugal blood flow of 235.32 +/- 56.24 ml/min, and the velocity of 8.62 +/- 2.33 cm/sec. Postoperatively, the gastric coronary vein was interrupted in 27 patients, the portal blood flow was significantly increased from 742.62 +/- 141.73 ml/min to 986.82 +/- 134.58 ml/min (P < 0.01) and the velocity was increased from 10.14 +/- 2.13 cm/sec to 13.41 +/- 1.91 cm/sec (P < 0.01). The results showed that the operation can stop abnormal blood flow in the gastrosplenic region and enhance liver perfusion. DCFI is useful for postoperative follow-up and observing portal hemodynamic changes.


[Old spinal cord injury treated by pulsed electric stimulation]. From March 1989 to May 1990, 80 cases of old spinal cord injury were treated with pulsed electric stimulation. The pulsed electric field was formed by utilizing paradural needle electrodes positioned cephalad and caudad to the site of spinal cord injury, with the stimulator connected with the electrodes being turned on. This stimulation was given 30 minutes once daily. 13 cases were treated for 3-6 months, 57 for 6-12 months, and 10 for 12-18 months. The nerve functions recovered in 78.5% of the cases (63/80), with the rate of useful recovery of 47.5% (38/80). The rate of EMG and gait improvement were 75.5% and 77.7%, respectively. The effect of pulsed electric stimulations was related to the extent of spinal cord injury, the depth and position of needle electrodes, the frequency and intensity of pulsed current and whether the spinal cord was compressed.


[Congenital coronary artery-cardiac chamber fistula: diagnosis and surgical treatment]. From Nov. 1976 to Dec. 1989, 9 patients with congenital coronary artery-cardial chamber fistula were operated upon in our hospital. In this series right coronary artery-right ventricle fistula was found in 3 patients, right coronary artery-left ventricle fistula in 2, left coronary artery-right ventricle fistula in 2, left circumflex artery-right atrium fistula in 1, and bilateral coronary artery-cardiac chamber fistula in 1. Cardiopulmonary bypass was performed in 8. The fistula was closed by interrupted mattress sutures outside the cardiac chamber plus ligation of terminal portion of the coronary artery near the fistula in 5 patients, and by mattress sutures inside the dilated coronary artery or cardiac chamber in other 4. There was no operative death in the 8 patients with single fistula. One patient with bilateral coronary artery fistula died from hypoxemia postoperatively. Late complications or recurrent fistulas were not found in the follow-up period from 3 months to 12 years. The diagnosis, operative indications and technique of closing fistula were discussed.


[A stereoscopic localization apparatus for early diagnosis of lung cancer and its clinical application]. We invented a stereoscopic localization apparatus for early diagnosis of lung cancer, facilitating fine needle aspiration biopsy especially for the deep-situated small lesion of the lung. Under X-ray, the stereoscopic localization of the three dimensions, posteroanterior sagittal and frontal plane, and transverse section of the body with small lesions can be soon established accurately with the apparatus. 227 percutaneous aspiration biopsies of lung lesions were performed by using the apparatus from 1985 to 1989. Of the 206 patients, 180 had lung lesions less than 3cm in size, and in 89.6% of the patients positive diagnosis was obtained by the first puncture biopsy. Satisfactory specimens were obtained in 96.6% of the biopsies. 26 malignant lesions about 1cm in diameter were localized, punctured and diagnosed accurately. They were subsequently confirmed surgically and pathologically. Mild pneumothorax occurred in 19 patients, and it subsided spontaneously.


[Surgical therapy of temporal lobe epilepsy. Clinical report of 30 cases]. Resection of epileptic foci for the treatment of temporal lobe epilepsy was performed in 30 patients (19 right, 11 left). Spike epileptiform wave over the temporal region was found in all patients. All the resected specimen were studied by light and electron microscopy. The operative results showed that no epileptic attack recurred after discharge in 10 patients; seizures were controlled by drugs after 1-2 attacks in 12 patients; rare or occasional attacks in 6 patients; and no effect after operation in 2 patients. The effective rate was 93%, and no death and severe complications occurred.


[Biomechanical and biological bases of the third type of fracture healing]. Fracture healing may take place in the form of direct repair by laminar bone with a little callus, and it is called the third type of fracture healing or an optimal form of fracture healing. The results of stress analysis of electric measurements on the femoral shaft after osteotomy, which is fixed with plates, in simulation to load tests demonstrated that if the design of the plate is reasonable, even the load is added over twice of the body weight, the compression stress subjected to osteotomy site may be greater than that or nearly the same as that in the intact bone. In another animal experiment, a model of the third type of fracture healing of the canine tibial shaft osteotomy fixed with trapezoid compression plate (TCP) was established with polychromatic sequential labeling. The results of 3-point bending tests indicated that the strength and the rigidity of the healing bone may be greater than normal region. The mechanical and biological bases of the third type of fracture healing are provided by these experiments.


Insulin antibody does not cause insulin resistance during glucose clamping in rats. Although it has often been stated that insulin antibodies cause insulin resistance, this concept is still controversial. The effect of insulin antibody GP30, commonly used in insulin radioimmunoassay, on insulin action was investigated in Wistar rats in vivo by the euglycemic glucose clamp technique. As a preliminary experiment, the equilibrium time required for insulin antibody to bind with endogenous insulin was examined. One hundred microliters/kg insulin antibody took 60 min or more to attain equilibrium, but 10 microliters/kg insulin antibody almost immediately equilibrated with endogenous insulin. During a 60-min glucose clamp study, 2 mU/kg/min porcine insulin was infused with 100 microliters/kg insulin antibody. At steady state, during the last 20-min period, the mean glucose infusion rate was 2.10 +/- 0.85 mg/kg/min (n = 5, mean +/- SD), significantly lower than the 5.77 +/- 1.61 mg/kg/min of the control, indicating insulin resistance before equilibrium was reached. However, the glucose infusion rates during the clamp with 10 microliters/kg insulin antibody and 100 microliters/kg insulin antibody infused 75 min before the insulin were 6.10 +/- 1.44 and 7.12 +/- 1.19 mg/kg/min, respectively, no different from the control. In these instances, free insulin levels measured by radioimmunoassay using the polyethyleneglycol method were 43.8 +/- 20.4 and 15.4 +/- 6.1 microU/ml, respectively, lower than the control (77.0 +/- 16.1 microM/ml).(ABSTRACT TRUNCATED AT 250 WORDS)


Increased oxidized form of human serum albumin in patients with diabetes mellitus. High-performance liquid chromatographic (HPLC) analysis of human serum albumin (HSA) on Asahipak GS-520H columns at neutral pH (6.87) showed a clear resolution of human mercaptalbumin (HMA) and nonmercaptalbumin (HNA), which are reduced and oxidized form of HSA, respectively. We studied the conversion of HMA to HNA (mercapt-nonmercapt conversion) as an index of oxidative change of the tissues and organs in 28 normal subjects and in a total of 47 patients with non-insulin dependent diabetes mellitus (NIDDM). Mean (+/- SD) values of the HMA fraction of HSA, f(HMA), [HMA/(HMA + HNA)], was significantly lower in NIDDM patients than in normal subjects (0.63 +/- 0.067 vs 0.75 +/- 0.028, P < 0.001). It was lower in poorly controlled NIDDM patients (0.63 +/- 0.058, n = 20) than in well controlled NIDDM patients (0.67 +/- 0.032, n = 9) (P < 0.05). Plasma glucose values sampled on occasions including overnight fasting and postprandial ones (r = -0.441, n = 47, P < 0.01), but not plasma glucose values sampled on overnight fasting (r = -0.345, n = 29) or postprandial (r = -0.467, n = 18) conditions and HbA1c (r = -0.211, n = 34), negatively correlated with the f(HMA) values, indicating that mercapt-nonmercapt conversion may not be due to cumulative hyperglycemia over a month, but due to short-term alteration in blood glucose level. The presence or absence of diabetic complications including nephropathy, retinopathy and neuropathy did not affect the f(HMA) values. In conclusion, decreased f(HMA) values in the diabetic patients suggested the presence of a rapidly altered oxidative change of albumin due to hyperglycemia.


Similarity of the constituents between glomerular arteriolar and mesangial lesions in experimental diabetes. In our previous studies in experimental diabetic rats, we have observed close similarities of ultrastructure and accumulation of IgG and IgM between the mesangial expansion and arteriolar hyalinosis of the glomerulus, and have presumed that both diabetic lesions are essentially of similar nature. In the present study, we carried out a further study on the constituents of both these lesions, using the PA-TCH-SP-PD technique for neutral carbohydrates, sialic acid and glycoproteins and the IgG-gold-silver technique for type IV collagen and fibronectin. The above staining and immunolabelings proved to be comparable in both lesions of diabetic glomerulopathies. This argues for the hypothesis of the identity of the two lesions.


A cross-sectional evaluation of cardiovascular risk factors in coronary heart disease associated with type 1 (insulin-dependent) diabetes mellitus. The contribution from lipoproteins, blood pressure, albuminuria and demographic variables to coronary heart disease in 90 adult subjects with and 172 without Type 1 diabetes mellitus was examined in order to investigate whether risk factors were of equivalent importance in diabetic and non-diabetic coronary heart disease. Coronary heart disease (CHD) was present in roughly 25% of subjects in each group. In Type 1 diabetes those with CHD had significantly higher levels of systolic blood pressure, albumin excretion, serum creatinine, triglycerides, VLDL cholesterol and C-peptide, and reductions in serum concentrations of HDL and HDL2 cholesterol, in comparison to those without. However, the prevalence of smokers, and concentrations of Lp(a), ApoB and fibrinogen were comparable. Blood pressure and HDL cholesterol were higher in the CHD group with Type 1 diabetes in comparison to the nondiabetic group with CHD, although LDL concentrations and the prevalence of Lp(a) concentrations > 200 mg/l were lower. Logistic regression analysis revealed the strongest independent predictors of CHD in Type 1 diabetes were serum triglycerides, systolic blood pressure, age, serum LDL cholesterol, and the daily insulin dosage, whereas in the non-diabetic control group HDL2 cholesterol, Lp(a), ApoA1 and ApoB, total serum cholesterol and body mass index were additional predictors. CHD in Type 1 diabetes appears to be most closely associated with increasing age and levels of blood pressure and total serum lipids. Apolipoproteins and albuminuria did not seem to be important independent predictors of CHD in Type 1 diabetes, whereas the former were more clearly associated with CHD in non-diabetic controls.


Morning or bed-time insulin with or without glibenclamide in elderly type 2 diabetic patients unresponsive to oral antidiabetic agents. We studied in a group of elderly (mean age 77 yr) non-obese Type 2 diabetic patients (n = 9) in a randomised, placebo-controlled prospective cross-over study of 8 months duration, the effects of substituting maximal sulfonylurea medication with a single injection of human zinc insulin taken either at bedtime (BTI) or morning (MI). All patients were poorly controlled with oral antidiabetic agents. After a 2-month regimen with either BTI or MI, a glibenclamide (GL, 3.5 mg/day) was given for an additional 2 months. Both insulin regimens decreased mean diurnal blood glucose and glycosylated HbA1c values to a similar extent (2.6-2.7%; p < 0.01-0.05), but with a lower daily insulin dose with BTI (0.30 +/- 0.03 IU/kg) as compared with MI (0.39 +/- 0.05 IU/kg; p < 0.01). A further improvement in metabolic control was observed in both groups after the introduction of GL; the mean reduction in glycosylated HbA1c was 1.4% for BTI and 0.7% for MI (p < 0.01 and 0.05, respectively), In conclusion, a subgroup of poorly controlled elderly Type 2 diabetic patients showed an improvement in metabolic control after a single injection of insulin despite discontinuation of maximal doses of oral antidiabetic agents. After 2 months of insulin treatment, a further improvement was achieved by a low dose of sulfonylurea in these patients who were formerly considered unresponsive to oral antidiabetic agents.


Epidemiology of childhood diabetes in Osaka District, Japan, using the documents from the medical benefits system specific for childhood diabetes. For an epidemiological study of childhood diabetes, application forms for medical benefits filed in Osaka Prefecture were collected and analyzed. A total of 465 patients with childhood diabetes under 18 yr of age were identified during the period between 1978 and 1988; 371 (175 boys and 196 girls) of them were classified as insulin-dependent diabetes mellitus (IDDM), and the others were either non-insulin-dependent diabetes mellitus (NIDDM) or an unidentified type of diabetes. The mean annual incidence of IDDM was 1.68 for boys and 2.00 for girls per 100,000 population, the rate being slightly higher for girls. The rates increased among older children, with a peak in the 10-14-yr age group. The prevalence estimated on the basis of the cumulative incidence in 1988 was 1.29 for boys and 1.60 for girls per 10,000 population, indicating a marked increasing trend with age. A significant seasonal variation was observed in the months of onset for IDDM, with higher rates during the winter. The mean birth weights of the male and female patients with IDDM were slightly lower than the national means. Use of official documents obtained from the medical benefits system was found to be an effective method for identification of cases of IDDM in the community, although further information is required for more accurate confirmation of such cases.


Therapeutical concentrations of tolbutamide, glibenclamide, gliclazide and gliquidone at different glucose levels: in vitro effects on pancreatic A- and B-cell function. In the classical model of isolated perfused rat pancreas four commonly used sulfonylureas--tolbutamide, glibenclamide, gliquidone and gliclazide--were investigated at therapeutical concentrations at three different glucose levels (with 0, 2.22 and 5 mmol/l glucose surrounding) and in the presence of a metabolic stimulus with glucose at 8.33 mmol/l. All the sulfonylureas stimulated the B-cell function. Tolbutamide, gliquidone and gliclazide produced a prompt biphasic hormone release while glibenclamide induced a delayed monophasic insulin secretion. In all cases the amount of insulin released depended on the metabolic condition. As the environmental glucose levels fell, the sulfonylureas' stimulatory effect on the B-cell function decreased. At the therapeutical concentrations we tested, no sulfonylurea influenced A-cell activity whether directly or indirectly via an insulin-mediated paracrine inhibition of glucagon release.


Changes in urinary retinol binding protein excretion and other indices of renal tubular damage in patients with non-insulin dependent diabetes. Changes in urine retinol binding protein (RBP, M(r) 21,000) excretion and other indices of renal tubular damage were investigated in the patients with non-insulin dependent diabetes mellitus (NIDDM). Changes in urine RBP excretion were well paralleled with those of urine NAG excretion. In RBP-negative patients, the subjects with hypertension (systolic blood pressure > or = 140 mmHg or diastolic blood pressure > or = 90 mmHg) showed higher beta 2-microglobulin (beta 2-MG) excretion and albumin (Alb)/Cr ratios than normotensive ones. In addition, both urine beta 2-MG excretions and Alb/Cr ratios were significantly increased in RBP-positive patients. The measurement of urine RBP excretion may have an additional role in the diagnosis of renal tubular dysfunction in diabetic patients.


[Administration of metoprolol in refractory and severe heart failure]. 46 cases of severe heart failure refractory to digitalis glycosides, diuretics and/or vasodilators were studied. First dose of beta-loc (metoprolol) was 6.25 mg twice a day. If no untoward effects, according to patient's condition, the dose was gradually increased to 12.5-25 mg b.i.d. for 4-6 weeks. Of the total 46 cases, improvement was found in 44, among which 23 cases (50%) were excellent and 21 cases (46%) fine. The effective rate reached 96%, which was very satisfactory.


[The effects of nitroglycerin on refractory congestive heart failure by intravenous infusion]. Noninvasive hemodynamic and clinical studies were carried out in 24 patients with congestive heart failure refractory to digitalis, diuretics and vasodilators. Nitroglycerin was given by intravenous infusion for 5-7 days. The results showed that the symptoms, signs and hemodynamic parameters were markedly improved. The overall clinical effective rate was 96% and no serious side effect was noted.


[Cardiopulmonary exercise test for evaluating cardiac reserve in chronic congestive heart failure]. To evaluate whether maximal oxygen consumption (VO2 max) measured by cardiopulmonary exercise test (CAR-PET) reflects cardiac reserve in patients with congestive heart failure (CHF), supine bicycle CAR-PET and exercise hemodynamic measurements were performed simultaneously in 12 patients with CHF of NYHA II-IV. With increasing workload, VO2 and cardiac output elevated gradually, then plateaued, demonstrating that patients with CHF could reach VO2max. According to VO2max, patients were divided into 4 classes: including 2 patients of class A (VO2max: 24.5 +/- 2.29 ml.min-1/kg, mean +/- s mean), 3 of B (17.6 +/- 1.37 ml.min-1/kg), 5 of C (13.6 +/- 0.66 ml.min-1/kg) and 2 of D (6.5 +/- 1.64 ml.min-1/kg). Maximal cardiac indices were 8.79 +/- 2.35 L.min-1/m2 in class A, 5.82 +/- 0.97 L.min-1/m2 in B, 3.53 +/- 0.95 L.min-1/m2 in C and 2.21 +/- 1.56 L.min-1/m2 in D. No significant correlation between supine resting hemodynamic parameters and VO2max/kg was found, suggesting that exercise tolerance could not be predicted by the measurement of resting cardiac performance. Furthermore, VO2max correlated poorly with NYHA classification in these patients. However, cardiac output correlated linearly with VO2 during exercise, suggesting that VO2 max/kg is a good predictor for cardiac reserve in CHF(CI = 0.6809 +/- 0.2748 VO2/kg, n = 40, r = 0.84, P < 0.0001; CO = 1.1618 +/- 7.9065 VO2, n = 40, r = 0.84, P < 0.0001). The results also showed that VO2max/kg did not correlate with the changes of pulmonary capillary wedge pressure (PCWP), indicating that exercise tolerance in CHF depends more on cardiac output than on ventilatory consequence of pulmonary congestion.(ABSTRACT TRUNCATED AT 250 WORDS)


[Catheter closure of patent ductus arteriosus without thoracotomy]. Transcatheter plugging of patent ductus arteriosus with modified Porstmann's method was performed in children of school age and adults (130 cases) in recent 10 years. We devised a set of instrument, self-made plastic Ivalon for tailoring the plugs, developed the technique of retrograde aortic angiography to measure the diameter of the ductus, and determined the criteria of definite lodgement of the plug. 3 cases failed because their femoral arteries were too small to have the catheter sheath introduced. The other 3 cases developed thrombosis of the femoral arteries after the procedure, and the thrombi were removed by surgery.


[Echocardiographic diagnosis and significance of atrial isomerism]. Forty-nine patients with atrial isomerism (right atrial isomerism 27, left atrial isomerism 22) diagnosed by 2DE of the abdominal great vessels were reported in this paper. Compared with the results of high kilovolt filtered chest film and autopsy, atrial situs determined by 2DE was concordant with bronchial morphology in 43 patients, 25 (93%) of right atrial isomerism, 18 (82%) of left isomerism. 25 patients who had angiocardiography or autopsy were associated with other cardiac malformations which were severe and complicated in right atrial isomerism. The results of this series showed that ultrasonography of the abdominal great vessels could be a reliable guide to the diagnosis of atrial isomerism, 2DE diagnosis is useful for angiocardiography.


[Entrainment phenomenon of atrioventricular nodal reentrant and atrioventricular reentrant tachycardias]. Atrioventricular nodal reentrant tachycardia (AVNRT) and atrioventricular reentrant tachycardia (AVRT) were induced by esophageal pacing and their entrainment zones were examined in 20 patients. The results showed that the entrainment phenomenon, which was a common electrophysiological phenomenon in reentrant tachycardia, occurred during overdrive pacing in 19 cases (19/20). The entrainment zone was 10-70 (30.50 +/- 20.85, mean +/- s) ms. The tachycardias could not be interrupted by pacing within the entrainment zone, but they were terminated by burst pacing, with starting pacing cycle length shorter than the shortest entrainment cycle length.


Mink as a predictive model in toxicology. This paper reviewed the biomedical and toxicological database concerning the use of mink as a predictive model of human responses. It is concluded that substantial information exists on the mink genetics, physiology, metabolism, nutritional requirements, and susceptibility to infectious disease; and provides a foundation upon which interspecies extrapolation may be considered. In addition, information on the response of mink to several dozen toxic substances revealed that mink respond in a qualitatively and quantitatively similar manner to other more commonly employed species as well as humans. Our conclusion does not infer that mink should be used routinely in toxicological testing for estimation of human responses. However, it indicates that toxicological data from this species may be a useful complement in risk assessment processes based upon data obtained from traditionally employed models such as rats and dogs.


[Comparative pathogenicity tests of eight pigeon paramyxovirus 1 variant isolates in pigeons, turkeys and chickens]. Eight pigeon paramyxovirus-1 isolates which were isolated from diseased pigeons were comparatively tested for their pathogenicity in chickens, turkeys and racing pigeons. Intramuscular inoculation of all of the eight viruses resulted in all pigeons in clinical signs like polyuria, lameness of wings and in parts also in torticollis. Also, intravenously inoculated chickens developed distinct signs such as apathy, liquid droppings and in part also torticollis. Six of the eight PMV-1 isolates induced in turkeys similar signs as in chickens; inoculation of two isolates yielded no signs in turkeys. Legal sanitary consequences of the disease due to pigeon PMV-1 infection in chickens and in turkeys should be identical to that of velogenic Newcastle disease.


[The aerobic bacterial flora of songbird nests]. In the area around Ober- and Unterschleissheim, a medium term decrease in the singing bird population appears to be happening. Therefore, material from nesting-boxes (used for nesting or sleeping) was examined for bacterial contamination. Mainly gram positive bacteria were isolated which were considered to belong to the normal flora. The occurrence of Streptomyces spp. is described in the nesting material of Passeres probably for the first time. Enterobacteriaceae represent only a fraction of the total isolates, but are demonstrated relatively frequently in the nests of the Nuthatch (Sitta europaea) and nests of other non-identified bird spp., so that colonization of the intestinal tract with Enterobacteriaceae cannot be excluded. Surprisingly, bacteria of the aquatic habitats such as Alcaligenes, Bordetella, Aeromonas, Non-Cholera (NC)-Vibrio were isolated, although in small amounts. The occurrence of NC-Vibrio has not yet been described in singing birds. Nests with high numbers of gram negative rods were successful in most instances, therefore, there was no proof that aerobic bacteria are responsible for the decrease in the population.


[Immunizing against salmonella infections with live and inactivated vaccines]. A live attenuated auxotrophic S. typhimurium (S. tm.)-mutant was used by orally administration via drinking water several times during rearing, combined with 1- or 2-times parenteral injection of an autogenous S. enteritidis (S. e.)/S. tm.-oil emulsion vaccine. In a 8-month period, more than 500,000 birds were vaccinated. The vaccine was safe. Challenge test showed protection in the vaccinates and their offspring. The number of isolates in the farms detected by regular monitoring decreased. The protection of the live-culture mutant lasted about only 8 to 10 weeks. A. S. tm.-mutant more potent for chickens will be tested now. We consider vaccinations as one important factor in a salmonella control program, especially for commercial layers and broiler breeders.


Comparative investigation on different turkey rhinotracheitis (TRT) virus isolates from different countries. The present investigation was carried out to compare the antigenic relationship between TRT virus isolates from different countries. The obtained results showed that all virus isolates shared similar physiochemical properties. In virus neutralisation tests marked two way cross reactions between BUT 1 = 8544 (England), STG 761/88 and STG 854/88 (Germany) could be detected. On the other hand VCO 3 isolate (France) showed only partial reaction. Also the SDS-poly acrylamide gel electrophoresis (SDS-PAGE) profiles of three TRT viruses (one from England and two from Germany) were very similar, while the VCO 3 strain from France showed some variation.


Effect of sample selection from experimentally contaminated hatching eggs and freshly hatched chicks on Salmonella enteritidis detection rate. The conventional culture methods of hatching eggs using shell and/or egg contents for detection of salmonella organisms give mostly unsatisfactory results. The aim of the present study is to evaluate selection of other samples and techniques of culturing hatching eggs and freshly hatched chicks. This current study provides the best evidence of Salmonella enteritidis in artificially contaminated eggs (Layer type) by using enrichment broth in empty egg shell samples in comparison to culturing samples from yolk, albumen or from shell above the air cell (with the outer shell membrane). The isolation rates could be enhanced if empty egg shell was initially filled with Buffered Pepton Water as pre-enrichment broth. Examination of organs from freshly hatched chicks revealed that crop samples give mostly higher reisolation rates.


[The survival ability of salmonella, coccidia oocysts and ascarid eggs in laying hen feces from different housing systems]. The time of survival of Salmonella typhimurium, coccidia oocysts and ascaris eggs in manure of layer was determined in 5 different housing systems and 2 storing places for litter. The experiments were carried out in a stable of experimental station Frankenforst of the university of Bonn with a flock of 2200 hens. The effects of the environment conditions temperature, dry matter content, pH-value and intestinal microflora of the manure have also been studied. The time of survival was different depending on the housing system. A recovery of viable coccidia oocysts was possible after 13-370 days, ascaris eggs 53-347 days and Salmonella typhimurium 2-175 days. The tenacity of the investigated test organism mainly depend on the dry matter content of the manure. The longest period of survival of salmonellas was found in dry environment conditions, were as coccidia oocysts and ascaris eggs have been observed with the shortest period of survival. The possibility of the examined resistant parasite stages to develop was disturbed. Only few of them were able to develop and with a longer development time than those examined in the control suspension. The results of this study indicate that chicken manure, before using it in plant production, should be stored long enough to prevent men or animals from possible infections.


[The tenacity of Newcastle disease virus (LaSota) in the excrement of laying hens in different housing systems]. The survival time of NDV (LaSota) in the excrement of layers was in summer (winter) 22 and 18 days (26 and 36 days) in two cage houses, 14 and 18 days (36 and 33 days) in two floor-pen houses, as well as 8 days (54 and 68 days) in two dropping store places. By one week staying in battery cages and following the storage in dropping store place after 47 or 50 days NDV (LaSota) could not be reisolated. Besides environment factors, temperature, pH and dry matter was the thermic effect very significant. The comparison of the tenacity of NDV (LaSota) in different housing systems was besides of the quantitative determination of survival time supported through the application of a life time distribution test.


[Flock data reported from a poultry slaughter house (broilers)]. Six broiler flocks have been compared with regard to the occurrence of attributes according to an examination protocol. Approved birds were scrutinized per random sample, condemned birds altogether. Parameters of examination were body weight occurring attributes (computed single and cumulated) count of dead and condemned birds in fattening period, transport and on slaughter line as well. Each flock was conspicuous in one or another respect. The graduation of the flocks was possible using the factors losses in the fattening period count of condemnation losses in the total production line coefficient of variation of weights attributes occurring in the meat inspection cumulated count of all attributes leading to condemnation. Results demonstrate, that graduation of different flocks by means of quantitative measurements is possible.


[Nitrogen and chemical oxygen demand burden of waste water caused by trout raising influenced by the protein content of the feed]. A possibility was shown, how to quantify the water content of nitrogen and COD (Chemical Oxygen Demand) from intensive fish production, independent of flow rate and feeding time. The nitrogen excretion could be reduced 50% by feeding a protein reduced fish feed (A: 38.4% XP) compared with an fish feed B contending 47.9% protein (XP). The excretion-compartments were evaluated with the waste water parameter COD. Doing this, it could be shown that the COD input by feed A is reduced 20% in opposite to feed B. Further more the separation of fish faeces would achieve an COD reduction from about 50% to 70%.


[A model trial for prophylaxis in intensive fish culture]. Prophylactic measures concerning stock management, premises (indoor systems), water supply (tap water as make-up water in reuse systems), water quality control and management, fish feed and staff (short term quarantine after contact with other fish or equipment) helped to build up and maintain a disease free stock of rainbow trout (Oncorhychus mykiss) over a period of seven years in a time-limited trial. 60% of fertilized eggs survived, 3 fish were lost out of 1000 reared to pan-size without any medication. Although it must be kept in mind that the trial was a small size undertaking it can be shown that consequent prophylactic measures can prevent disease mediated losses of fish.


[The significance of the weaning age of young rabbits for the development of young animals and their food consumption]. The experiment reported compared the influence of weaning at 25, 28, 31 and 35 days of age on the development of liveweight and food consumption during the pre- and post-weaning period hybrid rabbits. Results of 44 does and 949 offspring showed that late weaning exerted a positive influence on live-weight development in comparison with already weaned kits of the same age. In all weaning age groups a depression in growth occurred shortly after weaning which was compensated thereafter. There was no significant difference between treatments with regard to total food consumption of does and offspring. This lack of difference was due to the high food intake of lactating does after weaning that surpassed maintenance requirements.


[Injection technique and arthrography of the shoulder, elbow and carpal joints of the cat]. Nine cats were available for the present trial. Six thoracic limbs were dissected without any previous treatment. In the remaining twelve thoracic limbs, each articular space was injected with silicone and then fixed with a 5% formalin solution before being submitted to dissection. For the radiological examination ten adult cats were available and arthrographs were taken in two axial planes of each joint. The elbow-joints of six additional cats were injected with six differently formulated contrast media. Radiographs were taken at different time intervals: instantly, five and ten minutes after intraarticular injection. The quality of the contrast and clarity of the structures are discussed. A lateral and a cranial location for injection are described for shoulder-joint, as well as a caudolateral and a craniolateral location for the elbow. A dorsal injection location is recommended for the antebrachiocarpal articulation and the middle carpal articulation. The injection sides described are discussed comparing them with those taken from the literature on dogs and cats. The results obtained from arthrograms are compared with data from literature on dogs. The radiographs of the elbow show clear differences in quality depending on the contrast medium used and on time between injection and exposure. As a result, recommendations can be made for suitable contrast media for arthrograms of the shoulder-, elbow- and carpal-joint in cats. Further, it is recommended that radiographs should be taken not later than five minutes after injection of the contrast media.


[The arthrographic presentation of the hip, knee and tarsal joints of the cat]. The purpose of the present paper was to find out the most suitable localizations to puncture of hip, stifle and hock joints of the domestic cat. These joints are arthrographically presented. Further six contrast mediums in normal commercial usage were tested with regard to there contrast intensity. The only recommendable localization to arthrocentesis of the hip joint is at the cranial border of the greater trochanter of the femur. The stifle joint is approachable either at the medial or lateral border of the Lig. patellae. To puncture the proximal row of the hock joint the recommendable puncture site was found distal of the lateral malleolus between the tendons of the lateral digital extensor muscle and m. extensor digitorum longus. Any arthrocentesis of the distal row of the hock joint isn't recommendable. Contrast mediums with a iodine concentration below 300 mg/ml were not sufficient for arthrography.


Cranial and trunk neural crest cells use different mechanisms for attachment to extracellular matrices. We have used a quantitative cell attachment assay to compare the interactions of cranial and trunk neural crest cells with the extracellular matrix (ECM) molecules fibronectin, laminin and collagen types I and IV. Antibodies to the beta 1 subunit of integrin inhibited attachment under all conditions tested, suggesting that integrins mediate neural crest cell interactions with these ECM molecules. The HNK-1 antibody against a surface carbohydrate epitope under certain conditions inhibited both cranial and trunk neural crest cell attachment to laminin, but not to fibronectin. An antiserum to alpha 1 intergrin inhibited attachment of trunk, but not cranial, neural crest cells to laminin and collagen type I, though interactions with fibronectin or collagen type IV were unaffected. The surface properties of trunk and cranial neural crest cells differed in several ways. First, trunk neural crest cells attached to collagen types I and IV, but cranial neural crest cells did not. Second, their divalent cation requirements for attachment to ECM molecules differed. For fibronectin substrata, trunk neural crest cells required divalent cations for attachment, whereas cranial neural crest cells bound in the absence of divalent cations. However, cranial neural crest cells lost this cation-independent attachment after a few days of culture. For laminin substrata, trunk cells used two integrins, one divalent cation-dependent and the other divalent cation-independent (Lallier, T. E. and Bronner-Fraser, M. (1991) Development 113, 1069-1081). In contrast, cranial neural crest cells attached to laminin using a single, divalent cation-dependent receptor system. Immunoprecipitations and immunoblots of surface labelled neural crest cells with HNK-1, alpha 1 integrin and beta 1 integrin antibodies suggest that cranial and trunk neural crest cells possess biochemically distinct integrins. Our results demonstrate that cranial and trunk cells differ in their mechanisms of adhesion to selected ECM components, suggesting that they are non-overlapping populations of cells with regard to their adhesive properties.


Differential screening of a PCR-generated mouse embryo cDNA library: glucose transporters are differentially expressed in early postimplantation mouse embryos. Differential screening of a cDNA library constructed using PCR amplification techniques from RNA isolated from the distal portion (embryonic ectoderm, mesoderm and visceral endoderm) of 7.5 days post coitum (dpc) mouse embryos led to the isolation of two cDNA clones expressed at higher levels in 7.5 dpc embryos than 12.5 dpc embryos. Nucleotide sequence analysis revealed that each of these clones was a different member of the family of facilitative glucose transporters (Glut genes). The differentially expressed cDNA clones represent mouse Glut-1 and Glut-3. Levels of the Glut-3 mRNA declined 14-fold between days 7.5 and 12.5 of gestation, and were under our limits of detection by 14.5 dpc. The levels of the Glut-1 mRNA declined about 3-fold between days 7.5 and 12.5 of gestation. Analysis of the expression of these genes by in situ hybridization revealed striking differences in transcript localization in early postimplantation mouse embryos. At 7.5 dpc, both transporters were expressed more strongly in extraembryonic tissues than in the embryo proper. While both transporters were expressed in the amnion and chorion, only Glut-1 was expressed in the ectoplacental cone. In the yolk sac, Glut-3 appeared to be expressed only in the endoderm while Glut-1, although expressed in both layers, was expressed more strongly in the mesoderm layer. Thus, the two transporters have relatively reciprocal sites of expression in the developing extraembryonic membranes. Expression of Glut-1 was fairly widespread in the embryo at 8.5 dpc, but by 10.5 dpc expression was down-regulated and was observed in the eye and the spinal cord.(ABSTRACT TRUNCATED AT 250 WORDS)


Restoration of hair growth by surgical implantation of follicular dermal sheath. The capacity of lower follicle dermal sheath to restore hair growth was tested by removing the lower halves of follicles, and then immediately implanting material containing dermal sheath cells from these bases, into the remaining upper epidermal follicle cavity. Over 60% of recipient follicles produced stout emergent vibrissa fibres and some operations resulted in multiple hair production from a single follicle. Histological examination revealed new dermal papillae within large bulb structures which were sited below the level of amputation--a feature that indicated that the new dermal papilla was derived from implanted material. For many follicles, the failure to produce emergent fibres could be accounted for after histological examination. These results provide clear evidence that lower follicle dermal sheath cells are capable of replacing those of the dermal papilla and it shows that they can do so in the context of the upper follicle. However, because elements of lower follicle epidermis were present in the implant material, the interactive sequence of events cannot be established. Dermal sheath cells have immense potential for papilla cell replacement: questions remain as to whether the distinction between sheath and papilla cells is one of context, or whether the transition requires specific external influences.


Expression of Pax-3- and neuroectoderm-inducing activities during differentiation of P19 embryonal carcinoma cells. A P19 embryonal carcinoma stem cell line carrying an insertion of the E. coli LacZ gene in an endogenous copy of the Pax-3 gene was identified. Expression of the Pax-3/LacZ fusion gene in neuroectodermal and mesodermal lineages following induction of differentiation by chemical treatments (retinoic acid and dimethylsulfoxide) was characterized using this line and is consistent with the previous localization of Pax-3 expression in the embryo to mitotically active cells of the dorsal neuroectoderm and the adjacent segmented dermomyotome. Pax-3/LacZ marked stem cells were also utilized as target cells in mixing experiments with unmarked P19 cells that had been differentiated by pretreatment with chemical inducers. Induction of beta-galactosidase and neuroectodermal markers in the target cells demonstrates that: (1) some differentiated P19 cell derivatives transiently express endogenous Pax-3- and neuroectoderm-inducing activities, (2) undifferentiated target stem cells respond to these activities even in the presence of leukemia inhibitory factor and (3) the endogenous activities can be distinguished from, and are more potent than, retinoic acid treatment in inducing neuroectoderm. These observations demonstrate that P19 embryonal carcinoma cells provide a useful in vitro system for analysis of the cellular interactions responsible for neuroectoderm induction in mammals.


The expression of tissue and urokinase-type plasminogen activators in neural development suggests different modes of proteolytic involvement in neuronal growth. Tissue and urokinase-type plasminogen activators are serine proteases with highly restricted specificity, their best characterised role being to release the broad specificity protease plasmin from inactive plasminogen. It has frequently been suggested that these, and similar proteases, are involved in axonal growth and tissue remodelling associated with neural development. To help define what this role might be, we have studied the expression of the plasminogen activators in developing rat nervous tissue. Urokinase-type plasminogen activator mRNA is strongly expressed by many classes of neurons in peripheral and central nervous system. We have analysed its appearance in spinal cord and sensory ganglia, and found the mRNA is detectable by in situ hybridisation very early in neuronal development (by embryonic day 12.5), at a stage compatible with it playing a role in axonal or dendritic growth. Tissue plasminogen activator mRNA, on the other hand, is expressed only by cells of the floor plate in the developing nervous system, from embryonic day 10.5 and thereafter. Immunohistochemical and enzymatic analysis showed that active tissue plasminogen activator is produced by, and retained within, the floor plate. A mechanism is suggested by which high levels of tissue plasminogen activator produced by the stationary cells of the floor plate could influence the direction of growth of commissural axons as they pass through this midline structure.


Multiple gap junction genes are utilized during rat skin and hair development. The expression of four different gap junction gene products (alpha 1, beta 1, beta 2, and beta 3) has been analysed during rat skin development and the hair growth cycle. Both alpha 1 (Cx43) and beta 2 (Cx26) connexins were coexpressed in the undifferentiated epidermis. A specific, developmentally regulated elimination of beta 2 expression was observed in the periderm at E16. Coinciding with the differentiation of the epidermis, differential expression of alpha 1 and beta 2 connexins was observed in the newly formed epidermal layers. alpha 1 connexin was expressed in the basal and spinous layers, while beta 2 was confined to the differentiated spinous and granular layers. Large gap junctions were present in the basal layer, while small gap junctions, associated with many desmosomes, were typical for the differentiated layers. Although the distribution pattern for alpha 1 and beta 2 expression remained the same in the neonatal and postnatal epidermis, the RNA and protein levels decreased markedly following birth. Hair follicle development was marked by expression of alpha 1 connexin in hair germs at E16. Following beta 2 detection at E20, the expression increased for both alpha 1 and beta 2 in developing follicles. A cell-type-specific expression was detected in the outer root sheath, in the matrix, in the matrix-derived cells (inner root sheath, cortex and medulla) and in the dermal papilla. In addition, alpha 1 was specifically expressed in the arrector pili muscle, while sebocytes expressed both alpha 1 and beta 3 (Cx31) connexin. beta 1 connexin (Cx32) was not detected at any stage analysed. The results indicate that multiple gap junction genes contribute to epidermal and follicular morphogenesis. Moreover, based on the utilization of gap junctions in all living cells of the surface epidermis, it appears that the epidermis may behave as a large communication compartment that may be coupled functionally to epidermal appendages (hair follicles and sebaceous glands) via gap junctional pathways.


Expression of the genes for TGF alpha, EGF and the EGF receptor during early pig development. Expression of mRNA for transforming growth factor-alpha (TGF-alpha), epidermal growth factor (EGF) and the epidermal growth factor receptor (EGF-R) during early pig development was evaluated by reverse transcription-PCR. In the unfertilised pig oocyte, maternal transcripts for EGF, but not for TGF alpha or the EGF-R, were detected. Pig conceptuses were analysed at days 7, 8, 10, 12, 15, 17, 18 and 22 of pregnancy. EGF-R mRNA was detected at all stages of conceptus development analysed. Interestingly, TGF alpha mRNA was expressed by the developing blastocyst only at days 8, 10 and 12 of pregnancy, with the highest levels apparent at day 10. In contrast, EGF mRNA was first expressed by the post-elongation conceptus at around day 15 of pregnancy with levels continuing to increase up to day 22. In the day-18 and day-22 conceptuses, this EGF message was shown to be primarily a product of the embryo-amnion and not the placental membranes. Furthermore, EGF was immunolocalised in the day-22 embryo to the developing lung bud, gut loop and amnion. In summary, the expression pattern of TGF alpha mRNA during early pig development is coincident with the onset of blastocyst elongation and suggests a possible role for TGF alpha during this period of cellular remodelling. The temporal and spatial expression of EGF mRNA and protein suggests a possible involvement for EGF in the establishment of the early organ systems.


Commitment along the dorsoventral axis of the sea urchin embryo is altered in response to NiCl2. Few treatments are known that perturb the dorsoventral axis of the sea urchin embryo. We report here that the dorsoventral polarity of the sea urchin embryo can be disrupted by treatment of embryos with NiCl2. Lytechinus variegatus embryos treated with 0.5 mM NiCl2 from fertilization until the early gastrula stage appear morphologically normal until the midgastrula stage, when they fail to acquire the overt dorsoventral polarity characteristic of untreated siblings. The ectoderm of normal embryos possesses two ventrolateral thickenings just above the vegetal plate region. In nickel-treated embryos, these become expanded as a circumferential belt around the vegetal plate. The ectoderm just ventral to the animal pole normally invaginates to form a stomodeum, which then fuses with the tip of the archenteron to produce the mouth. In nickel-treated embryos, the stomodeal invagination is expanded to become a circumferential constriction, and it eventually pinches off as the tip of the archenteron fuses with it to produce a mouth. Primary mesenchyme cells form a ring in the lateral ectoderm, but as many as a dozen spicule rudiments can form in a radial pattern. Dorsoventral differentiation of ectodermal tissues is profoundly perturbed: nickel-treated embryos underexpress transcripts of the dorsal (aboral) gene LvS1, they overexpress the ventral (oral) ectodermal gene product, EctoV, and the ciliated band is shifted to the vegetal margin of the embryo. Although some dorsoventral abnormalities are observed, animal-vegetal differentiation of the archenteron and associated structures seems largely normal, based on the localization of region-specific gene products. Gross differentiation of primary mesenchyme cells seems unaffected, since nickel-treated embryos possess the normal number of these cells. Furthermore, when all primary mesenchyme cells are removed from nickel-treated embryos, some secondary mesenchyme cells undergo the process of "conversion" (Ettensohn, C. A. and McClay, D. R. (1988) Dev. Biol. 125, 396-409), migrating to sites where the larval skeleton would ordinarily form and subsequently producing spicule rudiments. However, the skeletal pattern formed by the converted cells is completely radialized. Our data suggest that a major effect of NiCl2 is to alter commitment of ectodermal cells along the dorsoventral axis. Among the consequences appears to be a disruption of pattern formation by mesenchyme cells.


An X-linked human collagen transgene escapes X inactivation in a subset of cells. Transgenic mice carrying one complete copy of the human alpha 1(I) collagen gene on the X chromosome (HucII mice) were used to study the effect of X inactivation on transgene expression. By chromosomal in situ hybridization, the transgene was mapped to the D/E region close to the Xce locus, which is the controlling element. Quantitative RNA analyses indicated that transgene expression in homozygous and heterozygous females was about 125% and 62%, respectively, of the level found in hemizygous males. Also, females with Searle's translocation carrying the transgene on the inactive X chromosome (Xi) expressed about 18% transgene RNA when compared to hemizygous males. These results were consistent with the transgene being subject to but partially escaping from X inactivation. Two lines of evidence indicated that the transgene escaped X inactivation or was reactivated in a small subset of cells rather than being expressed at a lower level from the Xi in all cells, (i) None of nine single cell clones carrying the transgene on the Xi transcribed transgene RNA. In these clones the transgene was highly methylated in contrast to clones carrying the transgene on the Xa. (ii) In situ hybridization to RNA of cultured cells revealed that about 3% of uncloned cells with the transgene on the Xi expressed transgene RNA at a level comparable to that on the Xa. Our results indicate that the autosomal human collagen gene integrated on the mouse X chromosome is susceptible to X inactivation. Inactivation is, however, not complete as a subset of cells carrying the transgene on Xi expresses the transgene at a level comparable to that when carried on Xa.


The consequences of ubiquitous expression of the wingless gene in the Drosophila embryo. The segment polarity gene wingless has an essential function in cell-to-cell communication during various stages of Drosophila development. The wingless gene encodes a secreted protein that affects gene expression in surrounding cells but does not spread far from the cells where it is made. In larvae, wingless is necessary to generate naked cuticle in a restricted part of each segment. To test whether the local accumulation of wingless is essential for its function, we made transgenic flies that express wingless under the control of a hsp70 promoter (HS-wg flies). Uniform wingless expression results in a complete naked cuticle, uniform armadillo accumulation and broadening of the engrailed domain. The expression patterns of patched, cubitus interruptus Dominant and Ultrabithorax follow the change in engrailed. The phenotype of heatshocked HS-wg embryos resembles the segment polarity mutant naked, suggesting that embryos that overexpress wingless or lack the naked gene enter similar developmental pathways. The ubiquitous effects of ectopic wingless expression may indicate that most cells in the embryo can receive and interpret the wingless signal. For the development of the wild-type pattern, it is required that wingless is expressed in a subset of these cells.


A new member of the spectrin superfamily may participate in the formation of embryonic muscle attachments in Drosophila. Myotube migration and the formation of muscle attachments are crucial events for the proper development of muscle patterning in the Drosophila embryo. This paper describes the identification of a new embryonic muscle-specific protein, MSP-300, in Drosophila. This protein is initially expressed by muscle precursors at muscle-ectoderm and muscle-muscle attachment sites. As development continues, MSP-300 becomes associated with muscle myofibrillar network. Studies of the subcellular localization of this muscle-specific protein in primary embryonic cultured myotubes show that MSP-300 decorates actin filaments, and that it is specifically enriched in sites where actin microfilaments are linked to the plasma membrane. Migrating myotubes exhibit high levels of this protein at their leading edge while, in myotubes that have already developed sarcomeric architecture, the protein is localized mainly at the Z-discs. Sequence of a partial 3.9 kb cDNA clone and molecular analysis of the predicted protein sequence of this protein indicates that it encodes a high relative molecular mass protein (approximately 300 x 10(3), which exhibits at least five spectrin-like repeats. Several properties are shared by MSP-300 and members of the spectrin superfamily: it is associated with actin microfilaments, its sequence exhibits spectrin-like repeats and it is localized at sites where actin is linked to the plasma membrane. This protein could have a developmental role in the formation of muscle-ectoderm attachments and may be involved in myotube migration on the ectoderm.


Multiple cholinergic differentiation factors are present in footpad extracts: comparison with known cholinergic factors. Sweat glands in rat footpads contain a neuronal differentiation activity that switches the phenotype of sympathetic neurons from noradrenergic to cholinergic during normal development in vivo. Extracts of developing and adult sweat glands induce changes in neurotransmitter properties in cultured sympathetic neurons that mimic those observed in vivo. We have characterized further the factors present in the extract and compared their properties to those of known cholinergic factors. When assayed on cultured rat sympathetic neurons, the major activities in footpad extracts from postnatal day 21 rat pups that induce choline acetyltransferase (ChAT) and vasoactive intestinal peptide (VIP) and reduce catecholamines and neuropeptide Y (NPY) are associated with a soluble protein of 22-26 x 10(3) M(r) and a pI of 5.0. These properties are similar to those of ciliary neurotrophic factor (CNTF). Moreover, the purified fraction from footpads has ciliary neurotrophic activity. Antibodies to CNTF that immunoprecipitate all differentiation activity from sciatic nerve extracts, a rich source of CNTF, immunoprecipitate 80% of the cholinergic activity in the footpad extracts, 50% of the VIP and 20% of the NPY activities. Neither CNTF protein nor CNTF mRNA, however, can be detected in immunoblot and northern analysis of footpads even though both CNTF protein and mRNA are evident in sciatic nerve. CNTF-immunoreactivity is associated with a sparse plexus of sensory fibers in the footpad but not with sweat glands or the Schwann cells associated with them. In addition, in situ hybridization studies with oligonucleotide probes failed to reveal CNTF mRNA in sweat glands. Comparison of the sweat gland differentiation activity with the cholinergic differentiation factor from heart cells (CDF; also known as leukemia inhibitory factor or LIF) suggests that most of the cholinergic activity in foot pads is biochemically distinct from CDF/LIF. Further, antibodies that block the activity of CDF/LIF purified from heart-cell-conditioned medium do not block the ChAT-inducing activity present in footpad extracts of postnatal day 8 animals. A differentiation factor isolated from skeletal muscle did not induce cholinergic properties in sympathetic neuron cultures and therefore is unlikely to be the cholinergic differentiation factor produced by sweat glands. Taken together, our data suggest that there are at least two differentiation molecules present in the extracts and that the major cholinergic activity obtained from footpads is related to, but distinct from, CNTF. The second factor remains to be characterized. In addition, CNTF associated with sensory fibers may make a minor contribution to the cholinergic inducing activity present in the extract.


Characterization of a germ-line proliferation mutation in C. elegans. The C. elegans germ line is generated by extensive proliferation of the two germ-line progenitor cells present in newly hatched larvae. We describe genetic and phenotypic characterization of glp-4, a locus whose product is required for normal proliferation of the germ line. glp-4(bn2ts) mutant worms raised at the restrictive temperature contain approximately 12 germ nuclei, in contrast to the 700-1000 present in wild-type adults. The few germ cells present in sterile glp-4 adults appear to be arrested at prophase of the mitotic cell cycle. This cell-cycle disruption prevents the germ cells from entering meiosis and differentiating into gametes. Shifting sterile glp-4 worms to the permissive temperature enables their germ cells to undergo extensive proliferation and form gametes, demonstrating that the bn2-induced cell-cycle arrest is reversible and that proliferation and differentiation of germ cells can be uncoupled from development of the somatic gonad. The glp-4(bn2ts) mutation can be used to generate large populations of worms that are severely depleted in germ cells, facilitating determination of whether any gene of interest is expressed in the germ line or soma or both.


Expression of the zinc finger gene Gli3 is affected in the morphogenetic mouse mutant extra-toes (Xt). Genetic analysis and homology between the phenotypic alterations of the human Greig Cephalopolysyndactyly Syndrome (GCPS) and the mouse mutant extra-toes (Xt) have suggested a dominant mutation in the same gene of both species. Recently, the GLI3 gene, a member of the Krüppel-related zinc finger genes, has been proposed as a candidate gene for GCPS. We examined the expression of the mouse Gli3 gene in both Xt mutant animals and during normal mouse development. Northern and RNAase protection analysis of embryos revealed that Gli3 expression was reduced about 50% in heterozygous Xt/+ mice and completely absent in homozygous Xt/Xt mice. In addition, in situ analysis of wild-type mice documented Gli3 expression in the developing limb and brain, structures affected in Xt mutant mice. This pattern suggests an important function of the Gli3 gene during morphogenesis.


Regulation of expression of fibronectin and its receptor, alpha 5 beta 1, during development and regeneration of peripheral nerve. The extracellular matrix glycoprotein, fibronectin, is a potent promoter of peripheral neurite outgrowth. Interactions of peripheral neurons with fibronectin have been shown to be primarily mediated by the beta 1 class of integrin heterodimers. In the present study, we have examined the expression and regulation of fibronectin and its integrin receptor, alpha 5 beta 1, in developing and regenerating chick peripheral nerve. We show that fibronectin and alpha 5 beta 1 are expressed at comparatively high levels in developing nerve with alpha 5 beta 1 expression on axons and non-neuronal cells. With nerve maturation, both proteins are less prominently expressed and the cellular pattern of alpha 5 beta 1 expression becomes more restricted. Following lesion of mature nerve, both fibronectin and alpha 5 beta 1 are strongly induced with prominent expression of alpha 5 beta 1 on regenerating neurites and Schwann cells. The elevation in fibronectin levels in the regenerating nerve is highest in the vicinity of the lesion, an area undergoing extensive cellular remodeling including Schwann cell migration and growth cone extension. Our results suggest that fibronectin and its receptor, alpha 5 beta 1, may mediate functionally important interactions in the development and regeneration of peripheral nerve.


In vitro solubility of human pulp tissue in calcium hydroxide and sodium hypochlorite. The tissue solvent capacity of a 2% stabilized sodium hypochlorite solution (Milton) and a commercial calcium hydroxide solution (Calasept) was examined under in vitro conditions where autolyzed human pulp fragments weighing approximately 0.0065 g were immersed in these solutions at 37 degrees C for periods of up to 10 days. It appeared that sodium hypochlorite was able to dissolve half the volume of pulp tissue within 1 h and the remaining tissue after 2-2 1/2 h. Calcium hydroxide dissolved half the pulp volume within 2 h, whereas it took 1 week for the remaining tissue to dissolve. These findings support the use of sodium hypochlorite as an irrigation solution during canal preparation and calcium hydroxide as a canal dressing for the purpose of creating a canal free of pulp remnants before root filling.