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In response to a widespread belief within the legal system that cross-examination is instrumental in uncovering the truth, we examined the effect of cross-examination questioning on the reports of children who had-and had not-been coached to lie. A group of children, aged 6-11 years (N = 65), played three computer games with one of their parents. For half of the pairs, the parents-who acted as confederates-coached their children to make lies of commission concerning the occurrence of two target activities. For the remaining pairs, these two target activities actually occurred, and there was no coaching. Immediately afterwards, children were interviewed about the two activities. Those who-correctly or incorrectly-reported that both activities occurred were retained for the final sample (n = 56); these children were then interviewed again with both neutral questions and cross-examination-style challenges. Neither style of questioning elicited responses that discriminated between liars and truth-tellers: although the accuracy of children who were lying increased in response to cross-examination questions, the accuracy of truth-telling children saw a corresponding decrease. When asked neutral questions, children's responses tended to be consistent with their earlier responses, whether or not those responses were lies. These findings raise important questions about the function that cross-examination might serve in trials involving child witnesses. Copyright © 2016 John Wiley & Sons, Ltd., (Copyright © 2016 John Wiley & Sons, Ltd.)

Dynamic analysis of oxygen (O₂) has been limited by the lack of a real-time, quantitative, and biocompatible sensor. To address these demands, we designed a ratiometric optode matrix consisting of the phosphorescence quenching dye platinum (II) octaethylporphine ketone (PtOEPK) and nanocystal quantum dots (NQDs), which when embedded within an inert polymer matrix allows long-term pre-designed excitation through fluorescence resonance energy transfer (FRET). Depositing this matrix on various glass substrates allowed the development of a series of optical sensors able to measure interstitial oxygen concentration [O₂] with several hundred millisecond temporal resolution in varying biological microdomains of active brain tissue., (Published by Elsevier B.V.)

To facilitate in-depth hazard prediction models, we must understand the spectral properties of expulsion plumes from conventional weapon attacks. Precise data on the spectral absorption of three chemical weapon agent simulants, in the infrared regime, are required to properly determine the mass of simulant in expulsion plumes from field demonstrations and small scale tests. Data for triethyl phosphate (a Soman simulant), triethyl phosphite (a Sarin simulant), and tributyl phosphate (a VX simulant) are presented. A thermal evaporation cell was designed and built that incorporated features that are not commercially available.

We set out to evaluate salivary cotinine concentrations to judge tobacco smoke exposure among infants and children, and to examine the results in relation to age and wheezing. This was a case-control study of wheezing children (n = 165) and children without respiratory tract symptoms (n = 106) who were enrolled in the Pediatric Emergency Department at the University of Virginia. The age range of both wheezing and control patients was 2 months to 16 years. Questionnaires were combined with cotinine assays in saliva to evaluate exposure to environmental tobacco smoke (ETS) for each child. The prevalence of exposure to one or more smokers at home was high (68%); and 43% of the children enrolled were exposed to ETS from their mothers. According to the questionnaires, and after adjusting for age and race, a wheezing child in this study was more likely than a control to be exposed to at least one smoker at home (odds ratio = 1.9; 95% CI = 1.1-3.4). However, the odds of exposure to ETS from smoking mothers did not differ significantly between wheezing and control patients, and no significant association was found between the presence of wheezing and salivary cotinine levels. Among children exposed to ETS at home, cotinine levels were significantly higher in saliva from those under the age of two years, and from toddlers aged 2 and 3 years, compared to values from children over age 4 years. Moreover, the number of smokers in the home strongly influenced cotinine levels from children under age 4 years. In addition, higher cotinine levels were observed in saliva from children under age 2 years who were exposed to ETS from their mothers. Cotinine levels were similar and significantly correlated in paired samples of saliva and serum from children under 4 years of age (n = 54), (r = 0.92, P < 0.001). Based on information gathered from questionnaires, the results indicate that wheezing children were more likely than controls to be exposed to ETS at home. However, significant differences in ETS exposure between wheezing and control groups with respect to maternal smoke exposure or comparisons of salivary cotinine levels were not apparent. It was clear that determinations of salivary cotinine for monitoring the prevalence and intensity of household smoke exposure in this study were most valuable during the first 4 years of life., (Copyright 2000 Wiley-Liss, Inc.)

Background: The amount of allergen necessary to sensitise genetically "at risk" children is unclear. The relation between allergen exposure and asthma is also uncertain., Methods: To ensure a wide range of allergen exposures the data from case-control studies of asthma in children aged 12-14 years attending three schools in Los Alamos, New Mexico and Central Virginia were combined. Skin prick tests to indoor and outdoor allergens and bronchial hyperreactivity to histamine were assessed in children with and without symptoms of asthma. The concentration of mite, cat, and cockroach allergens in dust from the children's homes was used as a marker of exposure., Results: Three hundred and thirty two children (157 with asthmatic symptoms and 175 controls) were investigated. One hundred and eighty three were classified as atopic on the basis of allergen skin prick tests and 68 as asthmatic (symptoms plus bronchial responsiveness). The prevalence and degree of sensitisation to mite and cockroach, but not cat, was strongly associated in atopic children with increasing domestic concentrations of these allergens. Asthma was strongly associated with sensitisation to indoor allergens (p<10(-6)) and weakly to outdoor allergens (p = 0.026). There was an association between current asthma and the concentration of mite allergen amongst atopic children (p = 0.008) but not amongst those who were specifically mite sensitised (p = 0.16)., Conclusions: The domestic reservoir concentration of mite and cockroach, but not cat, allergen was closely related to the prevalence of sensitisation in atopic children. However, the prevalence of current asthma had a limited relationship to these allergen measurements, suggesting that other factors play a major part in determining which allergic individuals develop asthma.

This cross-sectional emergency department study of 70 wheezing children and 59 control subjects (2 mo to 16 yr of age) examined the prevalence of respiratory viruses and their relationship to age, atopic status, and eosinophil markers. Nasal washes were cultured for respiratory viruses, assayed for respiratory syncytial virus (RSV) antigen, and tested for coronavirus and rhinovirus RNA using reverse transcription-PCR (RT-PCR). Also evaluated were eosinophil numbers and eosinophil cationic protein (ECP) in both nasal washes and serum, along with total IgE and specific IgE antibody in serum. Respiratory viruses were detected in 82% (18 of 22) of wheezing infants younger than 2 yr of age and in 83% (40 of 48) of older wheezing children. The predominant pathogens were RSV in infants (detected in 68% of wheezing subjects) and rhinovirus in older wheezing children (71%), and both were strongly associated with wheezing (p < 0.005). RSV was largely limited to wheezing children younger than 24 mo of age, but rhinovirus was detected by RT-PCR in 41% of all infants and in 35% of nonwheezing control subjects older than 2 yr of age. After 2 yr of age the strongest odds for wheezing were observed among those who had a positive RT-PCR test for rhinovirus together with a positive serum radioallergosorbent testing (RAST), nasal eosinophilia, or elevated nasal ECP (odds ratios = 17, 21, and 25, respectively). Results from this study demonstrate that a large majority of emergent wheezing illnesses during childhood (2 to 16 yr of age) can be linked to infection with rhinovirus, and that these wheezing attacks are most likely in those who have rhinovirus together with evidence of atopy or eosinophilic airway inflammation.

Pseudomonas aeruginosa, which was resistant to a wide variety of antibiotics, became sensitive to several of these antibiotics when grown and tested at 46 degrees C. Cell wall antibiotics such as penicillin G and ampicillin were only effective when added to cells growing at 46 degrees C prior to a temperature shift to 37 degrees C. Antibiotics which penetrate the cytoplasmic membrane to express their inhibiting action present a pattern different from those which are active against the outer cell wall. In order that these compounds be effective, the permeability of the cytoplasmic membrane must be further altered with agents such as EDTA which allow the penetration of actinomycin D. Inhibitors of protein synthesis, such as streptomycin and chloramphenicol, have increased access to their sites of action in cells grown at 46 degrees C. Cells grown at 46 degrees C have 40% less lipopolysaccharide (LPS) than cells grown at 37 degrees C and the LPS aggregates were of large molecular size in cells grown at 46 degrees C. Growth at 46 degrees C affects the permeability properties of the outer cell wall more than the permeability properties of the cytoplasmic membrane and this was due, in part, to the selective release of LPS of LPS-protein complexes at elevated growth temperatures.

The mechanism of uptake of benzoic and 2,4-dichlorobenzoic acid (2,4-DCBA) by Alcaligenes denitrificans BRI 3010 and BRI 6011 and Pseudomonas sp. strain B13, three organisms capable of degrading various isomers of chlorinated benzoic acids, was investigated. In all three organisms, uptake of benzoic acid was inducible. For benzoic acid uptake into BRI 3010, monophasic saturation kinetics with apparent K(infm) and V(infmax) values of 1.4 (mu)M and 3.2 nmol/min/mg of cell dry weight, respectively, were obtained. For BRI 6011, biphasic saturation kinetics were observed, suggesting the presence of two uptake systems for benzoic acid with distinct K(infm) (0.72 and 5.3 (mu)M) and V(infmax) (3.3 and 4.6 nmol/min/mg of cell dry weight) values. BRI 3010 and BRI 6011 accumulated benzoic acid against a concentration gradient by a factor of 8 and 10, respectively. A wide range of structural analogs, at 50-fold excess concentrations, inhibited benzoic acid uptake by BRI 3010 and BRI 6011, whereas with B13, only 3-chlorobenzoic acid was an effective inhibitor. For BRI 3010 and BRI 6011, the inhibition by the structural analogs was not of a competitive nature. Uptake of benzoic acid by BRI 3010 and BRI 6011 was inhibited by KCN, by the protonophore 3,5,3(prm1), 4(prm1)-tetrachlorosalicylanilide (TCS), and, for BRI 6011, by anaerobiosis unless nitrate was present, thus indicating that energy was required for the uptake process. Uptake of 2,4-DCBA by BRI 6011 was constitutive and saturation uptake kinetics were not observed. Uptake of 2,4-DCBA by BRI 6011 was inhibited by KCN, TCS, and anaerobiosis even if nitrate was present, but the compound was not accumulated intracellularly against a concentration gradient. Uptake of 2,4-DCBA by BRI 6011 appears to occur by passive diffusion into the cell down its concentration gradient, which is maintained by the intracellular metabolism of the compound. This process could play an important role in the degradation of xenobiotic compounds by microorganisms.

Objective: To compare eosinophil counts and concentrations of eosinophil cationic protein (ECP) in serum and nasal wash fluid from wheezing infants and children with those from age-matched children without respiratory tract symptoms., Design: A case-control study of 71 children treated for wheezing and 59 control subjects in the University of Virginia Pediatric Emergency Department. The patients ranged from 2 months to 16 years of age. Eosinophil numbers and ECP concentrations were assessed in serum and nasal washes. Total serum IgE was measured and the radioallergosorbent test was used to measure IgE antibody to common inhalant allergens., Results: Among children less than the age of 2 years, markedly elevated levels of ECP (> 200 ng/ml) were measured in nasal washes from 9 (41%) of 22 wheezing patients and 1 (6%) of 17 control subjects (p < 0.03). None of these children had a positive radioallergosorbent test result for IgE antibody to common aeroallergens or a nasal smear containing 10% eosinophils. Few of the wheezing children under 2 years of age had either increased concentrations of total IgE or ECP in their serum or an elevated total blood eosinophil count. After the age of 2 years, the percentage of patients with nasal ECP levels greater than 200 ng/ml was also significantly higher in wheezing children than in control subjects (p < 0.001), and a positive correlation was observed between ECP concentrations in their nasal washes and other eosinophil responses (total blood eosinophil counts, serum ECP levels, and nasal eosinophil counts)., Conclusion: Increased concentrations of ECP were detected in nasal washes from wheezing infants and children, indicating that eosinophils may contribute to the pathogenesis of airway inflammation in some children who wheeze early in life.

Background: Our objective was to identify the allergens associated with asthma among schoolchildren in an area of the United States where dust mite growth is expected to be poor. Los Alamos, N.M., was chosen because it has low rainfall and is at high altitude (7200 feet) making it very dry. One hundred eleven children (12 to 14 years old) from the middle school who had been previously classified according to bronchial hyperreactivity to histamine (BHR) were studied., Methods: Sera were assayed for IgE antibodies to mite, cat, dog, cockroach, Russian thistle, and grass pollen, with both CAP system fluoroimmunoassay (Kabi Pharmacia, Uppsala, Sweden) and conventional RAST. Allergens were measured in dust samples from 108 homes with two-site assays for mite (Der p 1 and Der f 1), cat (Fel d 1), dog (Can f 1), and cockroach (Bla g 2)., Results: Concentrations of dog and cat allergens were elevated in almost all houses with pets but were also high in a significant proportion of the houses without pets. Levels of mite allergen were less than 2 micrograms/gm in 95% of the houses, and cockroach was undetectable in all but two of the houses. Among the 21 with BHR who had symptoms, 67% had IgE antibody to dog and 62% had IgE antibody to cat. For these allergens IgE antibody was strongly associated with asthma (p < 0.001). By contrast, the presence of IgE antibody to mite, cockroach, or grass pollen was not significantly associated with asthma., Conclusion: The high prevalence of IgE antibody to cat and dog allergens among these children is in keeping with the presence of cat and/or dog allergen in most of the houses. Furthermore, sensitization (as judged by IgE antibodies) to cat and dog allergens was strongly associated with asthma. On the other hand, no clear relationship was found between sensitization or symptoms and the current level of allergen in individual houses. The results show that in this mite-and cockroach-free environment sensitization to domestic animals was the most significant association with asthma.

A survey of the Middle School in Los Alamos, N.M., USA, identified 57 children with symptoms of asthma and 54 controls. Among these children, sensitization to cat and dog allergens was very strongly associated with bronchial reactivity and symptoms. In contrast, sensitization to mites, cockroach, or grass pollen allergens was not significantly associated with symptoms or bronchial reactivity. In keeping with these results, dust from houses in Los Alamos only contained high levels of cat or dog allergens.

The authors report the inter-interviewer reliability of two brief questionnaires developed to measure the effects of innovations in methadone maintenance. The instruments were designed to answer the research questions, but to intrude only minimally into the clinical assessment and treatment processes. The Initial Interview, completed at the time of admission, yielded information on 23 variables, and the Followup Interview, completed as soon as possible after the first anniversary of admission, yielded information on 20 variables. To assess reliability, a repeat interview was conducted by a different interviewer immediately after the first interview was completed. Repeat interviews were conducted with 19 subjects who completed the Initial Interview and 30 who completed the Followup Interview. Exact agreement was found in all the pairs of responses from the Initial Interview for 5 of the 6 categorical variables and 6 of the 17 quantitative variables. For the remaining 11 quantitative variables, the intraclass correlation coefficients ranged from .700 to .999. Exact agreement was found in all pairs of responses from the Followup Interview for 2 of the 4 categorical variables and 8 of the 16 quantitative variables. For each of the remaining categorical variables, the kappa statistic was significant (.73 and .49). For the remaining 8 quantitative variables, the intraclass correlation coefficients ranged from .750 to .999. The findings signify satisfactory interviewer reliability of the instruments. These brief instruments could easily be adapted for use in other treatment evaluation studies where brevity in data collection is considered desirable.

Asthma in children and young adults is strongly associated with immediate hypersensitivity to indoor allergens, notably those derived from the house dust mite. In addition, outdoor air pollution is considered to aggravate existing asthma. We investigated the prevalence of asthma and the pattern of allergen sensitization in a mite-free environment with low levels of outdoor air pollution. A total of 567 children aged between 12 and 14 attending Los Alamos Middle School, NM (altitude 7,200 feet) were screened using a respiratory questionnaire; 120 children (53 control children) underwent allergen skin testing and serum IgE measurement, and their bronchial reactivity to histamine was measured. Dust was collected from 111 homes and the level of indoor mite and cat allergen measured. The prevalence of respiratory symptoms was high (13%), and from the detailed testing it was estimated that 6.3% of the children had asthma (defined as symptomatic bronchial reactivity). Children with asthma had elevated IgE, 367 (179 to 755) versus 38 (23 to 61), and predominant sensitization to cat, 68 versus 20% (p < 0.001). A high number of households (77%) had a pet cat or dog. The concentration of mite allergen was very low (mean 0.18 micrograms Der p milligrams sieved house dust), whereas that of cat allergen was high in homes with a cat (80.8 micrograms Fel d milligrams) but also in homes with no cat (3.2 micrograms Fel d milligrams). The results show that in a mite-free environment with low levels of outdoor air pollution, asthma was still a major cause of morbidity among schoolchildren.(ABSTRACT TRUNCATED AT 250 WORDS)

The computer modem technology being developed by Datascope Corporation for the purpose of off-site intra-aortic balloon pump (IABP) monitoring was clinically evaluated in 30 patients requiring IABP support. The Datascope System 95 model IABP with built-in modem was used on all patients. Remote communications via a personal computer with modem were conducted under both routine and emergency settings. The Datascope PC IABP software was evaluated for its user compatibility, efficiency, diagnostic capability and overall usefulness as a clinical tool. During the eight month evaluation period, 87 remote communications were conducted for both routine and emergency IABP evaluation checks. Adjustments were recommended on 22 occasions relevant to balloon timing, trigger mode selection and augmentation volume settings. Eight communications were initiated in emergency settings due to a variety of patient conditions. Emergency intervention was successful in diagnosing and resolving critical situations including atrial arrhythmias, pacemaker timing, low cardiac output syndrome, loss of trigger source, catheter malpositioning and poor augmentation. The diagnostic capabilities and efficient means of data collection by the computer software provide the clinician with a valuable tool for routine IABP clinical monitoring, as well as emergency problem resolution.

An automated thermal desorption gas chromatography technique has been adapted to analyse traces of volatile compounds in proprietary food-wrapping films. Fourteen brands of polyvinylchloride film, seven brands of polyethylene film and one polyvinylidene chloride film were discriminated. Prior infrared analysis was used to identify the polymer type. The chromatograms showed minor changes in volatiles along the length of a roll of film and major changes in films exposed to daylight or in contact with cannabis resin.

In 1972 a preliminary report described the use of a large-bore (Nos. 12 and 16 Fr.) vinyl catheter, which contained a steel trocar within its lumen, in repair of cystourethrocele in 86 patients. Further experience in an additional 158 patients undergoing cystourethrocele repair, and a variety of other gynecologic procedures, is now evaluated. The prototype of a sharp, beveled, cutting-edge trocar tip proved to increase substantially the ease of insertion. Problems of design and production of the catheter were encountered and corrected. Two points of technique, (1) insertion of the catheter prior to the surigcal procedure and (2) use of the two-hand method, are advocated. The trocar catheter was found to be a simple, practical, and safe instrument for suprapubic bladder drainage.

Twenty-two cases of vaginal vault prolapse were managed by our service over the five-year period ending 12/31/86. Eleven cases were repaired by a transvaginal sacrospinous colpopexy and 11 by a transabdominal suspension. There were no differences in demographic data or preoperative symptoms between these groups. The vaginal procedure required less operative time, had less blood loss, averaged fewer hospital days and overall was more economical. The complication rates and follow-up results of the two techniques were comparable. The abdominal procedure is considered preferable for patients with specific indications for an abdominal approach. The vaginal procedure appears to be more appropriate when the abdominal approach is not otherwise indicated. The experience of the surgeon and individualization of the patient are important factors in choosing the best approach for the repair.

Lysozyme (EC 3.2.1.17) complexes with extracted Pseudomonas aeruginosa LPS in two distinct stages. The initial stage does not produce turbidity detectable by nephelometry (measured as nephelos units (N) per time) but does permit low-speed sedimentation of the lysozyme-lipopolysaccharide (LPS) complex. This association is 100% disrupted by the action of 0.1 M Mg2+. Monovalent cations at equal ionic strength to the Mg2+ concentration used for these studies failed to alter significantly the lysozyme-LPS complex, indicating that the role of Mg2+ was not strictly an ionic one. The study of lysozyme-LPS complexes may provide a model system for investigating in vivo protein-LPS interactions.

Adhesions are the leading cause of small bowel obstruction and a frequent cause of failure of infertility operations. Fibrinolysis is involved in the formation and resolution of adhesions. Although intravenous dextran (Macrodex) is known to augment intravascular fibrinolysis, the effects of intraperitoneal dextran (Hyskon) on fibrinolysis have not been extensively studied. A fibrin plate assay system was used to assess plasminogen activator activity of rabbit peritoneum and plasma after treatment with intraperitoneal or intravenous dextran 70. Hyskon significantly reduced the ability of severe trauma to depress plasminogen activator activity of visceral peritoneum and was capable of direct plasminogen activation. Untraumatized or minimally traumatized peritoneum was not affected, nor was plasminogen activator activity of plasma. Pulmonary effusions and perioperative deaths were significantly associated with the use of Hyskon.

An alkaline phosphatase mutant of Pseudomonas aeruginosa exhibiting both regulatory and catalytic changes was isolated. Under repression conditions (i.e. high inorganic phosphate (Pi)) the mutant culture produced an alkaline phosphatase (APase) displaying significant activity against both beta-glycerol phosphate (betaGP) and p-nitrophenyl phosphate (pNPP), while the wild type displayed no activity directed towards these substrates under the same conditions. In vivo, the mutant enzyme's ratio of specific activities was 45:1 in favour of betaGP versus pNPP, whereas this ratio was reversed to 1:9 betaGP versus pNPP for the same enzyme isolated from mutant cells. In addition, the kinetic parameters and stability requirements for the mutant-derived enzyme was altered in comparison with those of the wild type. A study of lipopolysaccharide (LPS) preparations from both the mutant and wild type indicated the mutant to be deficient in the core region of its LPS. The authors propose that the modifications in the catalytic activity of the mutant enzyme, demonstrated in vivo, are due to a change in the enzyme's microenvironment.

Glutaraldehyde, the biological fixative of choice in the cytochemical localization of the phosphatases, was investigated for its effects on Pseudomonas aeruginosa alkaline phosphatase. Comparative studies on the inactivation of alkaline phosphatase by glutaraldehyde showed significant differences when the purified protein was compared with whole, cell-bound enzyme. The effects of the reagent on the kinetics of the purified enzyme were studied and some conclusions drawn as to the mode of inactivation. The reaction of glutaraldehyde with the cell envelope of P. aeruginosa was also investigated, and it was found not to modify the extraction of lipopolysaccharides from the outer membrane. This study emphasizes the care that must be taken to interpret data, cytochemical or otherwise, obtained when glutaraldehyde is used as a fixative or cross-linking reagent.

Pseudomonas aeruginosa ATCC 9027 grew on 0.5% (v/v) hexadecane as a sole carbon source in a chemically defined medium which required the addition of Fe3+ and Ca2+. There was a variable and extended lag period before an active growth rate was attained. Visible light microscopic evidence revealed that the bacteria did not adhere to hexadecane droplets suggesting the absence of a bioemulsifier. When compared with glucose-grown cells, hexadecane-grown cells produced 75% less lipopolysaccharide (on a total protein basis); this lipopolysaccharide contained 30-40% less carbohydrate, yet 50-75% more 2-keto-3-deoxyoctonate. These chemical changes made the cell surface appear more hydrophobic when tested in a biphasic hydrophobicity index system. Electron microscopy of thin sections and freeze etchings revealed hexadecane-grown cells contained granules which were judged to be polyphosphate by energy dispersive X-ray analysis. There was no apparent major morphological envelope alteration within the two cell types.

A strain of Pseudomonas aeruginosa, which was resistant to 400 mug/ml of chloramphenicol (CM), was isolated. The generation time of the resistant strain was the same in the presence or absence of CM and similar to that of the parent strain growing in the absence of chloramphenicol. Resistance is eliminated by treatment with acridine dyes, mitomycin C, and sodium dodecyl sulfate, suggesting that resistance may be expressed by a plasmid. The resistant strain does not produce the pigment pyocyanine and the addition of pyocyanine to this strain eliminates the resistance factor. A strain sensitive to CM was isolated. This strain does not produce the enzyme acetyl CoA : chloramphenicol transacetylase whereas the resistant strain does. The sensitive strain accumulates 14C-CM at a greater rate and to a greater extent than the resistant strain grown in the presence of CM. The results suggest that the resistant strain inactivates CM by acetylation and, in.addition, develops a "permeability" barrier towards chloramphenicol.

Pseudomonas aeruginosa (ATCC 9027) releases four periplasm-located enzymes, i.e., ribonuclease (EC 3.1.4.22; EC 3.1.4.23), alkaline phosphatase (EC 3.1.3.1), cyclic-2', 3'-phosphodiesterase (EC 3.1.4.d), and 5'-nucleotidase (EC 3.1.3.5) into the medium during growth. Ribonuclease and alkaline phosphatase are classed as enzymes which are readily extracted by osmotic shock and spheroplast formation whereas cyclic-2',3'-phosphodiesterase and 5'-nucleotidase are classed as enzymes which are not readily extracted by these procedures. In view of the relative ease of extraction of the former enzymes it is suggested that the lattter enzymes, cyclic-2',3'-phosphodiesterase and 5'-nucleotidase, are bound and located in the periplasm in a manner different to ribonuclease and alkaline phosphatase.

A new pressure technique for treatment of vaginal agenesis and stenosis, with the use of the bicycle seat stool, specifically designed Lucite dilators, and the patient model, is described. Experience with this method in 24 patients over a 5-year period produced coital function in 10 of 12 patients with vaginal agencies and 10 of 14 patients and various types of stenosis. Favorable experience with this technique, as compared to a previous 5-year period, in which the majority of vaginal agenesis was managed by the Abbe-McIndoe operation, has led to reversal of primary therapy of vaginal agenesis and stenosis from a surgical to a nonsurgical approach.

Alkaline phosphatase (APase) of Pseudomonas aeruginosa exists primarily in the periplasmic region of the cell, i.e., between the cytoplasmic membrane and the outer tripartite layer. The enzyme is also found in the culture filtrate or associated with the outer layer of the cell wall. APase forms a complex with released outer cell wall material, and lipopolysaccharide (LPS) is associated with the complex. Since the enzyme was purified to homogeneity, it became desirable to determine whether complex formation with LPS , or the outer cell wall, affected any properties of the purified phosphatase. The ratio of activities of purified APase with p-nitrophenylphosphate and beta-glycerolphosphate as substrates is about 4:1. The ratio of activities with enzyme complexed with LPS is about 1:1. The energy of activation of sucrose or magnesium released enzyme is 9500 cal/mol whereas the values for purified enzyme plus LPS, purified enzyme, purified enzyme plus phosphatidylethanolamine (PE), and purified enzyme plus LPS plus PE range from 3400 to 8700 cal/mol. These changes occur in the physiological temperature range, 27 to 39C, of this organism. Sucrose-released enzyme in the presence of substrate is inactivated at 47C whereas pure enzyme plus substrate is affected at 41C. The addition of LPS, PE, or a combination of both increases the temperature of inactivation from 45 to 51C. The results suggest that certain properties of the purified enzyme differ from those of the enzyme released from whole cells by either sucrose or magnesium resuspension. The addition of cell wall components such as LPS and PE to purified APase restores these properties. The addition of cell wall components such as LPS and PE to purified APase restores these properties. The evidence suggests that artificial complex formation changes the environment of the enzyme protein such that the environment now resembles that which exists within the whole cell wall.

Neisseria meningitidis SD1C exhibited a low tolerance to penicillin G (0.03 microgram/ml). Loss of viability in the absence of polyvinylpyrrolidone-40 and horse serum was independent of the concentration of antibiotic above the minimum inhibitory concentration, whereas the rate of bacteriolysis was concentration dependent. Penicillin-induced lysis was a secondary event in this organism. At low levels of penicillin G, growth characteristics, i.e., absorbancy changes, respiratory rate, and uptake of Mg2+, appeared normal during the first 90 min in penicillin; however, viability dropped dramatically. Additionally, total cell numbers remained constant while cell mass continued to increase at a rate normal for the population. The increase in cellular mass in the absence of cell division could be observed microscopically. Only one ultrastructural change induced by penicillin correlated with the loss in viability: the loss in continuity of the outer membrane with the peptidoglycan but only at the site of septum formation. This lesion did not occur when cells were grown in media supplemented with the protective agents polyvinylpyrrolidone-40 and horse serum. Under these conditions of growth and with relatively high levels of penicillin, constant viability was maintained, but cell division no longer occurred. Cell populations treated with penicillin in the presence of the protective agents became increasingly more dependent on the presence of these agents for total viability even in the absence of penicillin in the culture.

High-level chloramphenicol (CM) resistant variants of Pseudomonas aeruginosa were isolated after culture of the wild-type (WT) strain in broth containing high concentration of the drug. These variants exhibit reduced ability to accumulate several amino acids. The extent of reduction in transport capacity is a function of the concentration of CM in which the variants are grown. Respiratory activity is not reduced in these strains. Amino acid uptake is not affected by the presence of CM during assay. An isogenic strain carrying a plasmid coding for CM resistance does not show this response to CM. Transport capacity is restored to the WT level in CM-sensitive revertants. These results suggest that the acquisition of CM resistance in P. aeruginosa is associated with a fundamental alteration in membrane permeability which is regulated by metabolism in the presence of the drug. The ramifications of this for the study of CM action and resistance are discussed.

The Ingram method for creating a neovagina in women with vaginal agenesis or stenosis by using an intermittent pressure technique was performed on 45 patients between 1975 and 1982. Poor patient compliance has been the major drawback of previously reported pressure techniques. To overcome this, the Ingram method employs a unique approach using Lucite dilators and a mobile bicycle seat stool. Extensive nursing participation and a successfully treated former patient used as a motivating role model have significantly contributed to the acceptance of this method. Thirty-seven of the 45 patients were able to experience coitus subsequent to treatment.

Twenty-nine years experience with 346 patients with invasive carcinoma of the vulva is presented. More than 90 per cent had squamous carcinoma. The primary mode of treatment was surgical. Two hundred and ninety-six patients were treated primarily with surgical treatment, 120 underwent radical vulvectomy and bilateral groin and pelvic lymphadenectomy, 133 had radical vulvectomy with bilateral groin dissection and 390 receiving nonradical procedures. Thirteen patients had radical operations plus pelvic exenteration for advanced disease. There were no intraoperative deaths, but 16 (5.4 per cent) died within 28 days of the operation. The uncorrected over-all five year survival rate was 66 per cent. In the presence of negative nodes, it was 83 per cent and with positive nodes, it was 38 per cent. Fifty per cent of those treated with exenteration are alive and disease-free at five years or more. Since one-third of the patients presented with advanced disease (Stages III and IV), earlier diagnosis and prompt referral must be encouraged to improve surgical results.

Pseudomonas aeruginosa grows at an apparent reduced rate at 46 C as compared with the rate at 37 C, when growth is measured as an increase in absorbance. Cells at 46 C are long, plasmolyzed, nonmotile filaments. The filaments contain phase-dark material that may be chromosomal in nature. When the 46 C culture is shifted to 37 C, the filaments fragment at polar ends after flagella form, and the final number of cells is equal to the number of chromosomal "packets" observed within the filament. The outer envelope of the filament appears to be structurally complete as determined by biochemical, thin section, and freeze-etch examination. When filaments are treated with lysozyme, they form large spheroplasts, suggesting that the outer wall and the cytoplasmic membrane are continuous within the filament. Filaments produce little or no periplasm-located alkaline phosphatase (APase), but activity appears immediately after a shift to 37 C. Cells grown at 37 C and shifted to 46 C remain as single, nonmotile, rods or doublets, and the APase formed at 37 C remains stable at 46 C. The addition of APase or inorganic phosphate is partially or completely effective as an inducer of filament fragmentation at 46 C. The results suggest that periplasm-located APase is an important enzyme in the final stages of cell division when P. aeruginosa is cultured on inorganic phosphate-limiting media.

Aberrant breast tissue may occur anywhere along the embryonic milk line. Only 17 instances of breast tissue of the vulva have been recorded in the English literature between 1900 and 1976. Review of these cases, as well as features of the patient herein reported, serve as a basis for presenting a clinical profile of this interesting entity, its potential complications, and a rationale for appropriate management.

A review of the developmental research and the clinical utilization of the nonstress test (NST) is presented. In contrast to the oxytocin challenge test (OCT), it is more acceptable to patients, takes less time, is easily reproducible and is less expensive. More important, there are no contraindicatioins to the nonstress test as a primary screening device. The salient points in performance of the nonstress test, developed by other investigators as well as the authors in a level III maternity center, are reviewed. Enhanced sensitivity and specificity of this method for fetal evaluation must be pursued. No one criterion for a reactive nonstress test has been universally accepted. Furthermore, which test (NST or OCT) is the first to determine fetal distress remains unresolved. The "standard of care" for bioelectrical evaluation remains pluralistic, and it should be utilized within a system that uses additional modalities of fetal assessment and sound clinical judgment.