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The numbers and kinds of fungi that are known to cause human mycoses have increased greatly in the past decade. We report the first case (to our knowledge) of disseminated Phialophora parasitica infection. The patient, a 92-year-old woman, had known chronic subcutaneous infection with P parasitica. The systemic nature of her disease was diagnosed at autopsy, when the organism was identified in cultures obtained from a thrombus within an abdominal aortic aneurysm and was also histologically demonstrated within sections obtained from the aneurysm wall.

An enzyme-linked immunosorbent assay (ELISA) kit for herpes simplex virus developed by Ortho Diagnostic Systems, Inc., was evaluated. In phase I experiments, 263 clinical specimens from genital lesions were extracted into serum-free medium and then tested by ELISA for herpes simplex virus antigen. The results were compared with those obtained by conventional viral culture. Of 83 specimens, 65 were positive by ELISA (sensitivity, 78.3%). In phase II experiments, 249 clinical specimens were tested for herpes simplex virus antigen in direct specimen and in cell cultures (MRC-5 and rabbit kidney) incubated for 2, 4, and 7 days. Of 63 specimens, 40 were positive by ELISA in the direct specimen (sensitivity, 63.5%), and by 7 days incubation, 100% of the cultures positive by viral cell culture were also positive by ELISA. The ELISA was reproducible, and when both the direct detection and amplification culture were used, the sensitivity of ELISA paralleled the diagnosis of herpes simplex virus infections by viral cytopathic effect.

The p-nitro-alpha-acetylamino-beta-hydroxypropiophenone (NAP) differential test for the identification of Mycobacterium tuberculosis recovered from clinical specimens was evaluated by two laboratories and found to be a rapid and accurate procedure with a specificity exceeding 99%.

A case of mixed acute leukaemia with T lymphoblastic, myeloblastic, and monocytic components is described. The use of immunological markers, ultrastructural morphology, cytochemistry, immunochemistry, and combined techniques, simultaneously detecting two markers in individual cells, made it possible to define the different blast cell populations.

A profile scoring system has recently been developed as a standardized method of early identification and severity assessment of pregnancy-induced hypertension (PIH). The purpose of this study is twofold: (1) to compare the PIH profile scores of patients demonstrating mild PIH without proteinuria with those of patients demonstrating mild PIH with proteinuria, and (2) to introduce the PIH profile graph as a standardized clinical assessment tool. Serial PIH profile data after 24 weeks' gestation from 46 term primigravid patients with mild PIH (group 1 = 19 patients without proteinuria; group 2 = 27 patients with proteinuria) were plotted on PIH profile graphs and compared. Profile scores from ten normotensive primigravidas at term served as controls. Serial profile scores for groups 1 and 2 showed similar patterns on the profile graph. Based on these data, we believe that PIH without proteinuria is clinically and biochemically a manifestation of true preeclampsia before the onset of proteinuria. Furthermore, the PIH profile graph allows one to identify subtle changes in the disorder and to anticipate the development of severe preeclampsia.

Reovirus mRNA synthesis in vitro by the virion-associated RNA polymerase was only slightly (10 to 15%) diminished in the presence of 2 mM S-adenosylethionine. However, methyl group transfer from S-adenosylmethionine (0.05 mM) to the 5'-terminal cap structure, m7GpppGm in this mRNA was markedly inhibited (80%) under these conditions. Replacement of S-adenosylmethionine by S-adenosylethionine (5 mM) yielded mRNAs containing mainly (70%) 5'-terminal e7GpppGe and e7GpppG, but some of the products were unalkylated (5'-GpppG, ppG). The ethylated mRNAs, but not the unalkylated molecules, bound to wheat germ ribosomes and were translated essentially as well as the corresponding methylated mRNAs in wheat germ extracts and in nuclease-treated rabbit reticulocyte lysates. Protein synthesis directed by ethylated mRNAs in wheat germ extract was 80% decreased by 0.1 mM m7GMP. Under conditions of limited initiation, methylated mRNA bound to wheat germ ribosomes preferentially as compared to ethylated mRNA. The results document for the first time the synthesis of ethylated mRNA and support the hypothesis that N7-alkylation of the 5'-guanosine in caps, rather than methylation itself, is important for the enhancing effect of cap on the initiation of eukaryotic protein synthesis.

Inosine 5'-triphosphate (ITP) can be utilized in place of guanosine 5'-triphosphate (GTP) for both the initiation and the elongation steps of reovirus transcription, resulting in the synthesis of mRNAs containing 5'-terminal m7IpppIm and internal pI. The apparent molecular weights of the I-substituted products were altered as a consequence of the absence of G-C base pairs and accompanying loss of ordered structure. The migration of I-substituted RNAs in agarose gels and glycerol gradients was similar to glyoxal-treated transcripts; i.e., it decreased 2-fold as compared to the corresponding untreated G-containing mRNAs. 7-Methylinosine-capped (m7I-capped), I-substituted transcripts readily attached to wheat germ 80S ribosomes. Unlike native G-containing mRNAs, they also formed heavier complexes that sedimented faster than 80S complexes even in the presence of the nonhydrolyzable ATP analogue AMPP(NH)P and elongation inhibitor sparsomycin. I-substituted molecules that were capped posttranscriptionally to form m7G-capped 5' ends yielded mostly 80S monosomes, consistent with a strong influence of 5'-terminal structure on initiation of translation. Under limited conditions of initiation, I-substituted RNAs outcompeted G-containing transcripts for ribosome attachment. Although the results are consistent with enhanced binding and freer movement of ribosomes on unstructured templates, synthesis of acid-precipitable polypeptides in wheat germ extract directed by I-substituted RNAs was 15-fold less than with G-containing mRNAs.

We examined the hypothesis that initiation of eukaryotic protein synthesis involves ATP-dependent melting of 5'-cap-proximal secondary structure in mRNA by eukaryotic initiation factors 4A and 4B. In reticulocyte lysate depleted of ribonucleoside triphosphates by pretreatment with hexokinase/glucose, initiation complex formation by native reovirus mRNA showed a strict requirement for ATP. The corresponding mRNA synthesized with ITP in place of GTP to minimize secondary structure also required ATP for binding to 40 S ribosomal subunits in complexes characteristic of initiation. In a partial reaction without ribosomes, purified eukaryotic initiation factors 4A and 4B bound and cross-linked to the capped 5'-end of oxidized mRNA. This interaction was ATP-dependent with inosine-substituted or bromouridine-containing reovirus RNAs as observed previously with native mRNA. The results indicate that if initiation involves ATP-dependent denaturation of mRNA, the effect must occur after initiation factor-mediated attachment of mRNA to the 40 S ribosomal subunit.

Eukaryotic mRNA cap binding proteins were purified from ribosomal salt wash in the presence of protease inhibitors by sucrose gradient sedimentation and m7GDP-Sepharose affinity chromatography. Rabbit reticulocyte and erythrocyte proteins with sedimentation constants of less than 6 S yielded a approximately 24,000-dalton cap binding protein. It stimulated capped mRNA translation in extracts of uninfected HeLa cells but did not restore capped mRNA function in extracts prepared from poliovirus-infected cells. Restoring and stimulatory activities both were associated with a larger, approximately 8-10 S complex that included the approximately 24,000-dalton polypeptide and several higher molecular mass components. The same two translational activities were also obtained in a slightly smaller approximately 5-7 S complex from uninfected HeLa cells but were absent from poliovirus-infected cell preparations.

Roots of decapitated maize seedlings (Zea mays L.) were exposed for 12 hours to 1.0 millimolar KNO(3) (98.5 atom per cent (15)N) in the presence and absence (control) of 0.1 millimolar p-fluorophenylalanine (FPA), an analog of the amino acid phenylalanine. FPA decreased nitrate uptake but had little effect on potassium uptake. In contrast, accumulation of both ions in the xylem exudate was greatly restricted. The proportion of reduced (15)N-nitrogen that was translocated at each time was also restricted by FPA. These observations are interpreted as indicating that synthesis of functional protein(s) is required for nitrate uptake and for transport of potassium, nitrate, and reduced-(15)N from xylem parenchyma cells into xylem elements. The effect of FPA on nitrate reduction is less clear. Initially, FPA limited nitrate reduction more than nitrate uptake, but by 8 hours the cumulative reduction of entering nitrate was similar ( approximately 35%) in both control and FPA-treated roots. A relationship between nitrate uptake and nitrate reduction is implied. It is suggested that nitrate influx regulates the proportion of nitrate reductase in the active state, and thereby regulates concurrent nitrate reduction in decapitated maize seedlings.

A variety of 5'-3H-methyl-labeled, oxidized viral mRNAs were used as probes for detecting in wheat germ initiation complexes proteins that interact with, and can be cross-linked to, the 5'-cap structure. A limited and reproducible set of specific proteins was obtained with the different mRNAs. The binding of these proteins to the 5'-end of mRNA apparently results in protection against nucleotide pyrophosphatase digestion of the cap even in initiation complexes in which the 5'-end is susceptible to pancreatic RNase digestion. Cross-linked proteins from mammalian initiation complexes comigrated with several of the subunits of similarly treated eIF-3. A model for cap binding protein interaction with mRNA cap during initiation of translation is suggested.

The optimal management of stage II carcinoma of the endometrium remains to be established. We reviewed our experience in treating 42 patients with stage II endometrial cancer by surgery, radiation, or combined radiation and surgery at the Hospital of the University of Pennsylvania. The overall 5-year survival was 47.6%. The 5-year survivals of patients treated by surgery only, radiation only, or combination radiation and surgery were 68.5, 36.5, and 46.1%, respectively, which were not significantly different. Histologic grade was found a significant prognostic factor but type of cervical involvement was not. Major complication rates were similar in each treatment group. We conclude that the majority of patients with stage II endometrial carcinoma are best treated by combination radiation and surgery, but in a select subset of patients, radical hysterectomy and lymphadenectomy constitute a reasonable treatment option.

A 13-year-old girl with signs and symptoms of an acute abdomen was found to have uterus didelphys, unilateral hematocolpos, ipsilateral renal agenesis and menses. A review of the literature in English revealed 115 reported cases. This complex congenital anomaly is seen most commonly in adolescents with dysmenorrhea of progressive severity, abdominal pain and a pelvic mass.

Capped ribopolymers lacking a sequence complementary to the common 3' end of the influenza virion RNA segments effectively stimulated transcription of these RNAs by the virion-associated transcriptase. Thus, stimulation of transcription results not from hydrogen bonding between the capped RNA and the 3' end of the virion RNA but presumably from a specific interaction of the capped RNA with protein(s) in the transcriptase complex. Although no specific nucleotide sequence was required for priming activity, capped mRNAs with diminished secondary structure were preferred as primers. Inosine-substituted or bisulfite-modified capped reovirus mRNAs were at least 3- to 5-fold more effective as primers than were the native capped mRNAs. On the other hand, inosine substitution or bisulfite treatment of the uncapped form of reovirus mRNAs converted them from essentially inactive species to potent inhibitors of the transcriptase reaction primed by either ApG or globin mRNA. These effects of reduced secondary structure also most probably reflect an interaction of the exogenous RNAs with transcriptase protein(s). The results obtained from screening a series of native uncapped ribopolymers were consistent with inhibitory activity requiring the absence of most hydrogen bonding in the ribopolymer and also suggested that specific structural feature(s) of the nucleotides in the chain were important.

Although a latex agglutination test (StaphAurex) and a hemagglutination test (Staphyloslide) correctly identified all strains of Staphylococcus aureus that were susceptible or had intermediate susceptibility to oxacillin, 17 of 73 (23%) and 18 of 73 (25%) strains of oxacillin-resistant S. aureus were not identified by StaphAurex and Staphyloslide, respectively. All strains not detected were resistant to trimethoprim-sulfamethoxazole and rifampin.

Influenza viral complementary RNA (cRNA), i.e., viral mRNA was radioactive when purified from the cytoplasmic fraction of cordycepin-treated canine kidney cells that were incubated with [methyl-3H]methionine during infection. Approximately 55 to 60% of the methyl-3H radioactivity was in internal N6-methyladenosine, a feature distinguishing this mRNA from those viral mRNA's that are known to be synthesized in the cytoplasm. The remaining methyl-3H radioactivity was in 5'-terminal cap structures that consisted of 7-methylguanosine in pyrophosphate linkage to 2'-o-methyladenosine, N6, 2'-O-dimethyladenosine, or 2'-O-methylguanosine. Methylated adenosine was the predominant penultimate nucleoside in caps, suggesting that cRNA synthesis in infected cells initiates preferentially with adenosine at the 5' end. In contrast to cRNA, influenza virion RNA segments extracted from purified virus contained mainly 5'-terminal ppA and no detectable cap structures.

Eukaryotic initiation factor 4A (eIF-4A) has been purified (to apparent homogeneity) from rabbit reticulocyte lysate. It is a single polypeptide accounting for at least 90% of the Coomassie blue staining material when subjected to gel electrophoresis in the presence of sodium dodecyl sulfate. The molecular weight was determined by gel electrophoresis under denaturing conditions at two different bisacrylamide to acrylamide ratios, by gel filtration under native conditions and by sedimentation equilibrium at three different protein concentrations. Additional physical properties of the polypeptide were also determined. In an attempt to characterize the function of eIF-4A, a protein specifically required for mRNA translation, an assay was developed which measures the protein-dependent retention of radiolabeled hemoglobin mRNA on nitrocellulose filters. These studies led to the discovery of an ATP-stimulated binding of mRNA which is dependent on the presence of eIF-4A and eIF-4B that also contains the 24,000-dalton cap binding protein. The reaction apparently requires ATP hydrolysis since a nonhydrolyzable analogue of ATP, adenosine 5'-(beta, gamma-imino)triphosphate, does not stimulate mRNA binding and GTP cannot substitute for ATP. In addition, ATP-stimulated binding of mRNA can be inhibited by an analog of the mRNA 5' terminus, m7GMP, suggesting recognition of the capped 5' end of hemoglobin mRNA. Consistent with this suggestion, ATP also stimulated the covalent cross-linking of eIF-4A and eIF-4B to the cap of oxidized reovirus mRNA, an interaction that was inhibited by m7GDP.

Benzotrichloride (BTC) is used in the synthesis of benzoyl chloride and benzoyl peroxide. Epidemiological data suggest that BTC is a human lung carcinogen. In the present study, BTC was evaluated for its ability to induce lung adenomas in strain A/J mice. Four groups of 15 male and 15 female A/J mice were injected i.p. with either tricaprylin or BTC in tricaprylin three times a week for 8 weeks. BTC groups received doses totaling 1440 mg/kg, 719 mg/kg or 287 mg/kg. The mean number of lung tumors per mouse was 127 87 +/- 5.81, 43 +/- 2.44, and 17.73 +/- 1.09 in the groups treated with either 1440 mg/kg, 719 mg/kg, or 287 mg/kg, respectively. Tricaprylin-vehicle controls had a mean number of 0.46 +/- 0.15 lung tumors per mouse. Therefore, BTC produced a significant (P less than 0.001) and dose-related increase in the lung tumor response when compared to tricaprylin controls and is a potent carcinogen in the strain A mouse lung tumor bioassay.

The objective of this study was to develop a standardized method of identifying fetal-pelvic disproportion by comparing fetal head and abdomen circumferences with the respective maternal pelvic inlet and midpelvis circumferences. Fetal ultrasound and maternal x-ray pelvimetry measurements were performed on 75 term pregnant women with indications for a trial of labor. By means of individually measured fetal and maternal pelvic diameters, circumferences of the fetal head, fetal abdomen, maternal pelvic inlet, and maternal midpelvis were computed. Based on four circumference differences between the fetus and maternal pelvis (fetal head--maternal pelvic inlet, fetal head--maternal midpelvis, fetal abdomen--maternal pelvic inlet, and fetal abdomen--maternal midpelvis), a fetal pelvic index number was derived from the sum of the two most positive values. Of the 27 patients requiring cesarean sections for failure to progress in labor, all but four had positive fetal pelvic index values (sensitivity = 85%). Of the 48 patients who were delivered vaginally, all but four had negative fetal pelvic index values (specificity = 92%).

The fetal-pelvic index was recently introduced as an accurate method of prospectively identifying the presence or absence of fetal-pelvic disproportion. In that report the ability to detect fetal-pelvic disproportion in patients with macrosomic fetuses was not specifically addressed. The objective of this study is to compare the efficacy of three methods used to identify the presence or absence of fetal-pelvic disproportion (the fetal-pelvic index, Colcher-Sussman x-ray pelvimetry, and estimated fetal weight greater than or equal to 4000 gm) in patients delivered of neonates weighing greater than or equal to 4000 gm after an adequate trial of labor (N = 34). Of the 18 patients requiring operative intervention, all but one had a positive fetal-pelvic index value (sensitivity = 94%). Of the 16 patients in whom vaginal deliveries were spontaneous, all but one had a negative value (specificity = 94%). On the basis of these data, the fetal-pelvic index is an accurate method of predicting the presence or absence of fetal-pelvic disproportion in patients delivered of neonates weighing greater than or equal to 4000 gm.

Fibronectin, a major component of the extracellular matrix and basement membranes throughout the body, is thought to maintain the integrity of both the reticulo-endothelial system and microvasculature. In this study, plasma fibronectin levels were assayed by nephelometry in nine pre-eclamptic gravid women, nine normotensive gravid women and ten non-gravid women. The mean plasma fibronectin level (+/-S.E.M.) in pre-eclamptic gravidas (1687 +/- 101 micrograms/ml) is significantly higher than that of either normotensive gravidae (1129 +/- 99 micrograms/ml) or non-gravid women (897 +/- 60 micrograms/ml). Although the mechanism for elevated levels of plasma fibronectin in patients with pre-eclampsia is not clear, it may serve as an early biochemical marker for this disorder.

Protein synthesis initiation factors prepared from rabbit reticulocyte and mouse ascites ribosomes were tested for the ability to crosslink to the 5' cap of mRNA. Crosslinking of one polypeptide of apparent molecular weight 24,000 was inhibited by the cap analogs, m7GMP and m7GDP, indicating a specific interaction with the cap. Although specific crosslinking of the 24,000 molecular weight polypeptide was found with eukaryotic initiation factor 3 and to a lesser extent with initiation factor 4B, both of these factors contained less than stoichiometric amounts of this polypeptide. The crosslinking method provides a highly sensitive and specific assay for cap-binding proteins and should facilitate their purification for functional studies.

The absorption and assimilation patterns of (15)NO(3) (-) supplied as the Ca(2+) and Mg(2+) salts to intact ryegrass (Lolium perenne) seedlings were compared. No statistically significant effect of ambient cation on the amounts of (15)NO(3) (-) absorbed was observed in the initial six hours, but during the subsequent six hours, absorption from Ca((15)NO(3))(2) exceeded that from Mg ((15)NO(3))(2).Lower rates of (15)NO(3) (-) assimilation were found in roots exposed to Mg((15)NO(3))(2) than in those exposed to Ca((15)NO(3))(2). It was proposed that Mg(2+) initiated a restriction in (15)NO(3) (-) reduction in roots, probably as a consequence of a Mg(2+)-induced physiological Ca(2+) deficiency. Lower (15)N translocation rates were also observed from Mg((15)NO(3))(2). These effects of Mg(2+) in depressing (15)NO(3) (-) assimilation and translocation occurred prior to an effect on (15)NO(3) (-) uptake.In shoots, larger amounts of reduced (15)N products occurred with Ca((15)NO(3))(2) than with Mg((15)NO(3))(2). It was concluded that this was due to enhanced translocation of (15)NO(3) (-) (and possibly its reduced products) in presence of Ca(2+) rather than to specific cation effects on (15)NO(3) (-) assimilation in the shoots.

Experiments with intact plants of Lolium perenne previously grown with (14)NO(3) (-) revealed significant efflux of this isotopic species when the plants were transferred to solutions of highly enriched (15)NO(3) (-). The exuded (14)NO(3) (-) was subsequently reabsorbed when the ambient solutions were not replaced. When they were frequently replaced, continual efflux of the (14)NO(3) (-) was observed. Influx of (15)NO(3) (-) was significantly greater than influx of (14)NO(3) (-) from solutions of identical NO(3) (-) concentration. Transferring plants to (14)NO(3) (-) solutions after a six-hour period in (15)NO(3) (-) resulted in efflux of the latter. Presence of Mg(2+), rather than Ca(2+), in the ambient (15)NO(3) (-) solution resulted in a decidedly increased rate of (14)NO(3) (-) efflux and a slight but significant increase in (15)NO(3) (-) influx. Accordingly, net NO(3) (-) influx was slightly depressed. A model in accordance with these observations is presented; its essential features include a passive bidirectional pathway, an active uptake mechanism, and a pathway for recycling of endogenous NO(3) (-) within unstirred layers from the passive pathway to the active uptake site.