The use of seclusion on an inpatient crisis intervention unit.

Because of the use of seclusion is controversial, a retrospective study of 50 patients was designed to examine how seclusion is actually being used on a short-term inpatient crisis intervention unit. It was found that 44 per cent of the patients were secluded during their stay. Neither sex nor race seemed to be a factor in whether a patient was secluded, but elderly patients and depressed patients were less likely to be secluded. Most of the seclusions occurred on the first day of hospitalization, with the four most common reasons being agitation, uncooperativeness, anger, and history of violence. The author speculates that the use of seclusion on the crisis unit is related to the characteristics of the patient population as well as to the short duration of patient stay.


Utilization review and resident education.

The reasons for psychiatric hospitalization are not always taught clearly and formally to psychiatric residents; screening criteria employed in utilization review can be a tool for helping residents gather data and make decisions about the admission or continued stay of patients. On the admitting and inpatient units of the Payne Whitney Psychiatric Clinic, residents use sets of criteria for admission and for continued stay, with accompanying guidelines and clinical examples, as part of their training. The authros outline principles used in developing the criteria, including the belief that they should be applicable independent of diagnosis. They suggest that in a teaching hospital screening criteria should be taken as standards, and that they should be developed as part of the educational program and only then introduced into utilization review.


Hospitals should finance medical office buildings.

After the decision to build a medical office building on the hospital campus has been made, a new set of considerations must come to the fore. Although the best decision may be for the hospital to undertake financing of the building, this decision must be made in light of a number of important financial and legal considerations.


Hospital energy conservation grants program now under way.

A three-year, $900 million grant program, a part of the recently passed National Energy Act, will make grant funds available to not-for-profit hospitals and schools to help identify, implement, and install energy conservation measures. The following article describes the basic mechanisms and procedures for gaining access to the grant funds, according to the proposed regulations.


Six steps ease town-gown battle for educators and practitioners.

During the past 30 years, a "town-gown" conflict between health administration practice and education has evolved that parallels the situation between medical education and practice. The conflict has come  about as a result of the change from practitioner-dominated education to education that is controlled by full-time university faculty. Six steps are proposed to bring the practitioners and the educators into harmony, using the medical education model as a base for professional preparation of master's degree graduates in health services administration.


Center provides emergency care without unneeded inpatient units.

When a study showed that a hospital had enough beds for its service area but that one growing segment of this area needed faster access to emergency care, the Ambulatory Care Center/Emergency Services System (ACCESS) was established. Through its program and facility, ACCESS provides immediate ambulatory and emergency care in its community and, through referrals to the hospital's ED, ensures the provision of more extensive emergency or inpatient care, as needed. Telecommunications, coordination with the county rescue services and the hospital's ED, and cooperation with local private practitioners are vital elements of the program.


Auditing dietetic services: first of a series.

Dietetic audits may be used to assess quality of care or to evaluate the efficiency of a food service. Problems can be clearly identified through an audit, and solutions can be found to improve service and nutritional care. An audit also provides a basis for dietitian accountability. In this first of a series of four articles on dietetic audits, the authors describe the basic auditing process.


Custodians climb career ladder through intensive training program.

The training program for custodial staff at the University of Houston provides continuing education and opportunities for advancement for minority women, who make up approximately 90 percent of the staff. The training and opportunities offered may be responsible for the low turnover and absenteeism in the custodial services department.


Planning law deadlines threaten state programs.

By the end of June 1979, many states must have certification-of-need programs in compliance with federal regulations or lose federal funding for them. By the end of September 1980, states must be in compliance or lose federal funding for all state health resource development programs. Because these steps appear to be too politically severe for HEW to implement, Congress will have to take steps in the 96th Congress to modify the impact of these deadlines.


Voluntary effort has goal in sight.

The success of the Voluntary Effort can be credited to the movement's broad-based coalition and the commitment shown at the grass roots level. But much remains to be done, and the VE is gearing up to meet the challenge. In 1979, emphasis will be placed on: a greater role and involvement for hospital trustees, increased efforts by hospitals to develop better ways to inform physicians about the costs of tests and services, and the need to create an even greater awareness and commitment to the VE at the grass roots level.


Operational audit can help ensure long-term cost containment.

Any health care institution can use operational auditing, or operational review, to regularly evaluate the performance of specific units or of the entire institution, to identify conditions that need the most improvement, and, thereby, to select the best cost containment and revenue-enhancing approaches. This article discusses certain features and advantages of operational auditing, such as its use of industry and other substantive standards and the auditors' independence from the institution's internal politics. It also explains some important differences between operational auditing and internal auditing and the details of the operational auditing process.


Affirmative action: what's really fair?

The concept of affirmative action has been the subject of much controversy in the past. Opponents of the concept believe that it provides preferential treatment for minorities and females based upon factors of race and sex that are both unconstitutional and illegal. Proponents believe the concept is needed to make up for past abuses. In this article, the author attempts to illustrate how an objective affirmative action program can enhance a hospital's personnel development system.


The President's NHI proposal: solution or problem?

Are the Carter administration's NHI principles appropriate for resolving health problems in the United States? An examination of the applicability of these principles to the problems--categorized in terms of health status indicators, health care resources, and consumer satisfaction--concludes that evidence in these areas does not indicate a need for NHI. Recommendations for establishing appropriate health care proposals follow.


The energy audit: back to the basics.

Gathering information about the cost of energy and patterns of usage within the hospital is the first step in mounting a program for conserving resources. A survey of the plant or a full-scale audit by an energy team can help administrators plan an energy-saving program.


Samaritan Health Center: an ecumenical merger.

The consolidation of a Protestant and a Catholic hospital has resulted in the first interfaith health care facility in the United States. While the obstacles to merger for hospitals with separate boards of directors and staff are many, the possibilities for success are enhanced by the Christian viewpoints the hospitals espouse.


Hospital-sponsored positive health programs.

Several kinds of positive health programs, ranging from efforts to identify cases of hypertension to the establishment of prenatal and well baby clinics, can be advantageous to the community hospital and to those it serves. Catholic hospitals, with their commitment to social justice, can use their unique positions as community centers of health care to pioneer the development of such programs.


The pathology of adrenocortical neoplasia: a correlated structural and functional approach to the diagnosis of malignant disease.

The structural and functional characteristics of nine functioning adrenocortical tumors (four adenomas and five carcinomas) causing Cushing's syndrome or virilization were studied. All tumors that we considered to show histologic evidence of malignant disease and that subsequently metastasized or recurred also showed in cell culture at least one significant functional or behavioral difference from benign tumors. No single defect was common to all carcinomas, but predominant changes included secretion of precursor steroids, such as 11-deoxycortisol (S) and a blunted or absent response to ACTH. All adenomase examined were normal in these respects in comparison with nondiseased cortical cells in culture. In carcinomas whose functions deviated only minimally from normal the presence of highly differentiated ultrastructural characteristics did not, however, confer a better prognosis.


The ultrastructure of bronchial macrophages and lymphocytes in sarcoidosis.

A physical interaction between macrophages and lymphocytes was observed more frequently in bronchial lavage fluid obtained from patients with sarcoidosis than from normal volunteers, irrespective of their history of smoking. In this spontaneous interaction more than two lymphocytes were commonly seen adhering to a macrophage without evidence of cytoplasmic bridging or membrane fusion. To a greater extent than in normal volunteers, macrophages from patients with sarcoidosis were characterized by the appearance of a more highly irregular cell surface, more membrane bound inclusions, however, was positively correlated with the smoking history of the individual, and the number of surface projections (microvilli) of macrophages from smokers appeared to be reduced. Significant differences were not apparent in the nuclear or cellular diameters of macrophages from sarcoid and normal individuals.


Crohn's disease: a scanning electron microscopic study.

A scanning electron microscopic study of Crohn's disease was done using surgically resected specimens. Grossly normal resection margins as well as nonulcerated portions from diseased areas were selected for study. Scanning electron microscopic findings in Crohn's disease included changes in villous size and shape, villous fusion and epithelial bridge formation, goblet cell hypertrophy and hyperplasia, and increased secretion of mucus. These changes were marked in involved areas, and many were also present in six of seven margins of resection available for study in the ileal group. The abnormalities found in grossly normal margins of resection suggest a more widespread involvement than can be appreciated by gross and light microscopic examination of the specimen. Formation of the increased coat of mucus observed may be stimulated by a number of agents and could contribute to an enhanced barrier function in areas of early involvement in Crohn's disease, thereby decreasing the uptake of toxic or antigenic macromolecules. A decrease in bacterial superinfections would also be facilitated. Later lesions, with severe villous changes and hypersecretion of mucus, may favor the uptake of toxic and antigenic macromolecules as well as aid in the establishment of bacterial superinfections.


Paragangliomas of the head and neck region. A pathologic study of tumors from 71 patients.

The histopathology of 72 paragangliomas originating in the head and neck region of 71 patients is presented. There were 45 carotid body, 13 vagal body, eight jugulotympanic, and three nasal paragangliomas. In addition, two arose in the larynx and one in the area of the aortic arch. Tumors occurred in four unrelated families. The two most important histologic features leading to a diagnosis of paraganglioma were zellballen and the presence of cytoplasmic argyrophil granules in all cases in which staining with the Grimelius technique was carried out. Ultrastructural study of three carotid body and two vagal body paragangliomas revealed both light and dark chief cells. Tumor cells contained membrane bound, electron dense neurosecretory types of granules, which usually ranged in diameter from 120 to 200 nm. Follow-up information was available for 67 patients (94 per cent). Two of the three nasal paragangliomas, 50 per cent of the jugulotympanic paragangliomas, 17 per cent of the vagal body paragangliomas, and 10 per cent of those of the carotid body recurred locally following attempted surgical resection. All patients treated with radiation had persistent tumor. Four (9 per cent) of the carotid body paragangliomas were malignant, all four patients dying with widespread metastases. One vagal body paragangliomas metastasized to regional lymph nodes (the patient was alive and well at five years), and another caused death by direct intracranial extension. In contrast to the benign tumors, malignant paragangliomas tended to show foci of necrosis and vascular invasion. Mitotic figures, which usually were not identified in the benign cases, were seen in all malignant tumors.


Sarcoidosis of the uterus.

Sarcoidosis rarely involves the female genital tract. This report describes a case of sarcoidosis of the uterus that was found by endometrial curettage for menometrorrhagia. Reports of the few previously documented examples of sarcoidosis of the uterus are reviewed with a brief discussion of the differential diagnosis.


Dysplasia of the kidneys, liver, and pancreas: report of a variant of Ivemark's syndrome.

A newborn girl with respiratory distress due to bilateral pneumothorax was found to be anuric, and died at 18 hours of age. Autopsy revealed a large pancreatic cyst, multiple large hepatic cysts, congenital hepatic fibrosis, bilateral dysplastic kidneys, and dysplasia of the pancreas. These findings constitute a variant of Ivemark's syndrome of dysplasia of the pancreas, liver, and kidneys.


Trisomy 10p due to a de novo t(10p;13p).

A new case of trisomy 10p has been identified by means of the GTG-banding technique. The patient is a female child carrying a sporadic translocation, t(10;13)(p11;p11), and affected by microsomatia and microcephaly with facial dysmorphia, retarded growth, weight gain, and psychomotor development, and bilateral talipes.


Trisomy 12p syndrome: de novo occurrence of mosaic trisomy 12p in a mentally retarded boy.

The first case of trisomy of probable 12p mosaicism originated de novo is presented. Comparison of the clinical findings of this patient with those of previously described cases of 12p trisomy derived from translocated chromosomes indicates that the symptoms of 12p trisomy are: (1) normal birth weight and physical development, (2) severe psychomotor retardation and generalized hypotonia, (3) peculiarly round face with prominent cheeks, hypertelorism, epicanthus, broad, flat nasal bridge, short nose with anteverted nostrils, large philtrum, broad, prominent lower lip, and (4) poly(syn)dactyly of feet.


A case of trisomy 3q21 leads to qter syndrome.

An infant with karyotype 46,XY,der(8),t(3;8)(q21;p23) is presented. The presence of trisomy 3q21 leads to qter syndrome is suspected on the basis of comparison of the clinical and laboratory findings of this patient with those of cases that have been reported as partial 3q trisomy. The common phenotypic features of this syndrome include growth failure and mental or developmental retardation, hypotonia, persistent lanugo, distorted head, congenital glaucoma, short and upturned nose, prominent maxilla, micrognathia, short, webbed neck, short limbs, retroflexed third and fourth toes, cutaneous syndactyly of the second, third and fourth toes, and elevated galactose-1-phosphate uridyl transferase activity in the red blood cells.


Translocation C:D involving chromosomes 11 and 14.

A translocation of material from chromosome 11 to chromosome 14 was identified in a 7-month-old male with microcephaly and developmental delay. The break-points appear to be on the long arm of chromosome 11, close to the centromere, and on the short arm of the 14.


An azoospermic male with a Y/autosome translocation.

This report describes an azoospermic male carrying a Y/autosome translocation. The patient had a 46,X,t(Y;10)(q12;p13) chromosome complement in a lymphocyte culture. The cytogenetic study of this patient is described, together with testicular histology, spermiogram, hormone levels, and clinical history.


A woman carrier of two apparently unrelated reciprocal translocations: prenatal diagnosis of normal karyotype in the foetus.

Two reciprocal balanced translocations involving chromosomes 2, 9, 12, and 18 were found in the karyotype of a woman with a child showing several congenital malformations at birth. Prenatal cytogenetic diagnosis, performed when a second pregnancy occurred, showed a normal chromosome constitution in the foetus.


Genetic linkage relations of the sixth component of complement (C6).

Linkage relations between the C6 and 33 other genetic marker loci have been analyzed in Norwegian pedigrees, including 114 matings with 388 informative children, by use of the MOSM computer program. No suggestion of linkage was found. Very close or close linkage (theta less than 0.06) has been ruled out for males between C6 and the following 19 marker loci: GPT, HLA + Bf, Rh, C3, Hp, PGM3, Km, Gm, Fy, Gc, ABO Jk, GLO1, K, MNSs, PTC, ACP1, PGM1, and Pi. For several of the relations even loose-linkage is unlikely.


Clinical and biochemical investigations on patients with partial deficiency of placental steroid sulfatase.

We report on three independent cases with a partial deficiency of placental steroid sulfatase (E.C.3.1.6.2). Upon routine pregnancy monitoring these patients were detected on the basis of low estriol excretion and failing induction of labor. In all three cases a male was delivered and subsequently the diagnosis of partial deficiency of placental steroid sulfatase was confirmed enzymatically in placenta homogenates. In one case, fibroblast cultures were established from skin explants of mother and son. In fibroblasts of the child, as in placental tissue, the activity of steroid sulfatase was only 34% of normal. Similar values were obtained for arylsulfatase C, though this enzyme is clearly separable from steroid sulfatase by electrophoresis. In cells of the mother, enzyme activities were unremarkable.


'GPI Roma', a new glucose phosphate isomerase deficient variant: in vivo occurrence of postsynthetic modifications of the mutant enzyme.

In a 5-year-old Italian girl with severe congenital hemolytic anemia, red cell GPI deficiency was proven, and found to be due to a new variant, 'GPI Roma.' The parents are first cousins and have been proven to be heterozygous for this variant. GPI Roma was slightly unstable to heat and exhibited a slightly increased Michaelis constant for fructose-6-phosphate. A single predominant fast-migrating GPI form existed in the patient's white blood cells, while the electrophoretic pattern in the red cells was composed, in addition to this 'fast band,' of a major band migrating as normal GPI and of an additional slow band. It is shown that this phenomenon may be ascribed to postsynthetic events modifying the charge of the mutant enzyme.


Occurrence of the ACP01 allele in Czechoslovakia.

During a paternity test an unexpected type of red cell acid phosphatase isozyme (ACP1) was found in one family. The mother was of type A and type B was diagnosed in the son. The whole family was then subjected to ACP1 phenotyping and to the enzyme assay. Five members of the family were found to have unexpected types of ACP1 isozymes. The average activity was approx. 50% of normal values. It is presumed that a silent ACP01 allele was found in the family investigated and that the grandfather was its first carrier.


Partial duplication of the short arm of chromosome 9 (p13 leads to p22) in a child with typical 9p trisomy phenotype.

A 3-year-old girl with duplication 9 (p22 leads to p13) is reported. The presence of a classical 9p trisomy phenotype in this patient suggests that this region (or part of it) is responsible for the major, typical clinical stigmata of this partial autosomal trisomy syndrome.


Mosaic 13 trisomy due to de novo 13/13 translocation with subsequent fission. Karyotype: 46,XX, - 13, + t(13;13)(p11;q11)/46,XX,del(13)(p11).

A severely retarded child with multiple malformations was found to present a mosaic karyotype 46,XX, - 13,+t(13;13)(p11;q11)/46,XX,del(13)(p11), which probably originated as the result of a de novo 13/13 translocation in a parental gamete, followed by postzygotic fission of the translocation chromosome.


Partial trisomy 14q.

A dysmorphic female born with partial trisomy of the proximal segment of the long arm of chromosome 14 had 47 chromosomes. The extra one was acrocentric, smaller than the D group, and bigger than the G-chromosome group. By GTG banding it was identified as a deleted chromosome 14, the karyotype being 47,XX,+del 14(q24). Chromosome analysis of the parents was normal.


IgE antibody response to mite antigens in mite infested mice.

Mice infested at birth with the mouse mite Myocoptes musculinus developed positive skin tests to mite antigens at the age of 5 weeks. Serum IgE antibodies directed against mite antigens were first detected at 6 weeks of age and high levels of IgE were present as long as 1 year later. Similar kinetics of IgE formation were observed in mice infected as adults. Mast cell degranulation by mite extract was demonstrated in connective tissue obtained from the skin of mite infested mice.


Production of xenoantiserum to a chemically induced murine tumor: analysis of tumor specificity in different immune tests.

The method of in vivo absorption of xenoantisera raised against murine chemically-induced tumors has been used to obtain specific anti-tumor sera. To assess if the tumor specificity was maintained under different experimental conditions, the sera were employed in in vitro tests based on different immunologic reactions. In vivo absorption results in the production of xenoantisera which are highly specific for the immunizing tumor and that lack antibody activity toward histocompatibility and organ specific antigens and endogenous virus antigens. Thus, in vivo absorbed antisera appear to be valuable reagents for the immunochemical characterization of chemically-induced tumors.


Correlation of tartrazine hypersensitivity with specific serum IgD levels.

The clinical diagnosis of hypersensitivity to the food dye tartrazine in 16 subjects correlated with levels of tartrazine-specific IgD antibodies determined by a solid phase radioimmunoassay inhibition test. This is the first study associating hypersensitivity to a low molecular weight chemical with an antibody response of the IgD class. There was little or no correlation between clinical sensitivity and IgE antibodies.


Attachment to erythrocytes of uniform salt forms of lipopolysaccharides from Salmonella abortus-equi and its inhibition by various animal sera.

Lipopolysaccharides (LPS) readily attach to erythrocyte membranes in vitro, resulting in hemagglutination in the presence of the homologous bacterial antibodies. The attachment of LPS to red blood cells can be prevented by certain, but not all, animal sera. In this study, the attachment of uniform salt forms of LPS from Salmonella abortus-equi to erythrocytes from various animal species was investigated. The uniform salt preparations were significantly more active than the starting material. Similar findings were obtained with erythrocytes from man, sheep, rabbit, and guinea pig. These results were not due to differences in antibody neutralizing capacity, since all preparations showed essentially identical activity. When sera from various animal species were used as inhibitors of LPS attachment, inhibition of erythrocyte modification by equivalent, but not by identical, amounts of LPS was essentially the same.


Multiple effects of a diamidine (propamidine) on complement activation.

Propamidine, one of the diamidines used against infections with babesiae has inhibitory and enhancing effects on complement activation as assessed by immune haemolysis of sensitized sheep red cells. Utilization of C1 is powerfully, that of C3 weakly improved by propamidine while activation and/or fixation of C4, C5 and to a lesser degree of C8 and C9 are inhibited. At low concentrations of propamidine (less than 2 mM) the enhancing effects, at higher concentrations the inhibitory effects predominate. Inhibition is produced, in some cases certainly, in others likely, by interference of propamidine with binding properties of complement components. None of the complement enzymes, C1s, C42 or C3bBb was inhibited in its hydrolytic activity. The possible significance of propamidine actions is discussed.


Interference of propamidine with binding of the fifth component of complement to surface-fixed C3b, and with C5 activation.

The effects of propamidine and their dose dependency, on utilization of the third and fifth complement components in immune haemolysis have been compared. While C3 utilization is not disturbed that of C5 is markedly inhibited by propamidine in concentrations as low as 0.5 mM. Both, binding of C5 to surface-fixed C3b and cleavage of C5 by convertases C42 and C3bBb, are also inhibited in the presence of propamidine. Since neither C3 cleavage by these enzymes nor even C5 cleavage by the cobra venom factor-supported convertase CFVBb is significantly reduced a general convertase-inhibiting effect of propamidine is ruled out. Rather the effect on utilization of C5 is the result of interference with binding of C5 is the result of interference with binding of C5 to C3b and hence impairment of its accessibility to the convertases. These findings thus further support the role of surface fixed C3b in C5 activation proposed earlier.


The effect of age on IgE production in rats.

Inbred BN-rats were immunized at different ages with a standardized amount of ovalbumin and a1(OH)3. The specific IgE antibody response and total serum IgE levels were significantly influenced by age. The IgE antibody response increased to a maximum in 2-month-old rats and decreased progressively at a more advanced age. Total serum IgE levels increased progressively from the age of 1 month to the age of 5 months and decreased again at the age of one year.


The effect of heat inactivation of serum on aggregation of immunoglobulins.

Heating serum at 56 degrees is used to inactivate complement in several immunological assays. During heating, both heat-labile and heat-stable anticomplementary activity (ACA) develop. While heat-labile ACA can be completely inactivated, heat-stable ACA increases progressively with continued heating. Heat-stable ACA develops in deaggregated IgG and in normal, but not in hypogammaglobulinaemic, human and porcine serum heated at 56 degrees suggesting that this ACA is due to formation of immunoglobulin aggregates. These aggregates would produce false-positive tests for immune complexes and could inhibit a variety of cell-mediated reactions in assays which incorporate heat-inactivated serum. Other temperatures were tested to determine whether endogenous haemolytic activity could be destroyed without forming immunoglobulin aggregates. At 53 degrees both endogenous haemolytic activity and heat-labile ACA were inactivated and formation of heat-stable ACA in normal serum was minimal. ACA, however, could be induced in deaggregated IgG at 53 degrees. Moreover, the degree of heat-induced aggregation of IgG in vitro at either temperature was directly proportional to IgG concentrations and inversely related to albumin concentrations. Thus, pathological sera with these protein alterations might form more aggregates during heating than normal sera. These data suggest the following: (1) heat inactivation of complement at 53 degrees for 90 min is preferable to the traditional 56 degrees; (2) in any assay where immunoglobulin aggregates might interfere, normal serum may be an inadequate control and correlations will need to be made between serum IgG and albumin concentrations and the results obtained in these assays.


Isolation and characterization of the third component of rat complement.

The third component of complement from rat plasma was isolated by precipitation with polyethyleneglycol in the presence of benzamidine, followed by chromatography of the precipitate on CM-cellulose, hydroxyapatite, and QAE A50-Sephadex, and gel-filtration on Sephadex G-200 superfine. C3 was isolated in its native form, as assessed by its specific functional activity and its immunoelectrophoretic mobility. The recovery of C3 was between 18 and 26%, and the final material was homogenous on SDS-PAGE analysis. Rat C3 has an apparent molecular weight of 187,000 and it is composed of two non-identical disulphide linked polypeptide chains of 123,000 and 76,000. A monospecific antiserum against rat C3 was produced by immunization of rabbits with purified C3. Zymosan treatment of rat serum resulted in cleavage of C3 and in the appearance of component with a greater mobility during immunoelectrophoresis. The plasma concentration of rat C3 in Wistar rats was 581 +/- 49 microgram/ml (mean +/- 1 S.D.). Interaction of highly purified rat C3 with purified human components B, D, and P resulted in the formation of a C3 convertase suggesting compatibility between human and rat C3.


Immunological unresponsiveness during induction of experimental autoimmune orchitis in guinea-pigs: studies in vivo and in vitro.

Groups of male and female guinea-pigs were immunized with homologous sperm derived from testis (TS) or epididyimis (ES) in Freund's complete adjuvant (FCA). In vivo investigations included skin tests at 2 weeks and development of aspermatogenesis (testis weight) at 4 weeks; in vitro assays were inhibition of migration of peritoneal exudate cells (PEC) and culture of blood leucocytes (lymphocyte transformation) at weekly intervals after immunization. Antigens used were heat-treated extracts of sperm used for immunization (BTS, BES); cells were also cultured simultaneously with PPD. Skin tests revealed anergy in males as compared with females: a larger quantity of antigen which caused partial unresponsiveness in females, caused profound unresponsiveness in males although the aspermatogenesis was less severe. In vitro tests also showed anergy during the active stages of the orchitis. This was non-specific for PEC (specific unresponsiveness was not excluded), but blood leucocytes showed only specific unresponsiveness (to BES). These and previous studies suggest that the unresponsiveness results from a desensitisation by sperm antigens released during the development of aspermatogenesis.


Effects of acquired resistance on infection with Eimeria falciformis var. Pragensis in mice.

Mice immunized with infections of 500, 5,000, or 20,000 oocysts of E. falciformis var. pragensis were reinfected with 20,000 and 100,000 oocysts at 20 and 38 days, respectively, after the initial infection. After the first challenge infection, none of the immunized mice showed clinical signs of coccidiosis; a few mice passed very low numbers of oocysts, and oocyst discharge seemed to correlate negatively with immunizing dose. None of the mice immunized twice passed oocysts after challenge. Mice immunized with three infections were completely immune to challenge for 4 months. The effect of the immune response on the life cycle of the coccidium was determined by histological examination of the intestines of immune and nonimmune mice infected with the parasite. In both the immune and nonimmune groups, sporozoites penetrated absorptive epithelial cells and migrated to crypt epithelial cells during the first 6 to 24 h postinfection. At 48 to 72 h postinfection, the sporozoites developed into mature first-generation schizonts in the nonimmune mice, whereas the developing first-generation schizonts degenerated within the crypt epithelial cells of the immune mice. In nonimmune mice, third-generation merozoites, inoculated intracecally, developed into mature fourth-generation schizonts, whereas in immune mice the developing fourth-generation schizonts degenerated before maturing. The possibility that a cellmediated immune mechanism is responsible for the arrest in schizogony is discussed.


Viral etiology of age-dependent polioencephalomyelitis in C58 mice.

The etiology of immune polioencephalomyelitis (IPE) and the mechanisms of resistance to IPE induction were investigated in C58 mice. IPE was found to be induced by a lipid-solvent-sensitive, filterable replicating agent present in line Ib leukemic cell suspensions. IPE was serially transmitted in immunosuppressed mice with filtered extracts of spleens from diseased animals. The IPE-inducing activity of Ib cell extracts was abolished by chloroform or deoxycholate. Gel filtration of Ib cell extracts showed that the IPE agent has a molecular weight of at least 10(7). Electron microscopy of the active fractions from columns and of spinal cord extracts from mice with IPE revealed a virus-like particle, 40 nm in diameter, which is probably the IPE revealed a virus-like particle, 40 nm in diameter, which is probably the IPE agent. Administration of cyclophosphamide at various times after challenge increased the incidence of IPE in mice, suggesting that IPE is not autoimmune mediated. Immunosuppression resulted in maintenance of high levels of IPE agent in the central nervous system tissue, while immunization resulted in low levels. Moreover, immunized mice produced neutralizing antibodies. These data suggest that antibodies help restrict the amount of IPE agent in the nervous tissue, and that this restriction is required for resistance to IPE induction in C58 mice.


Depression of cell-mediated immunity in cholera.

Cholera toxin may depress cell-mediated immunity by stimulation of adenyl cyclase and production of cyclic AMP in cellular systems or when given parenterally to experimental animals. Whether or not similar effects might be found during clinical infection with Vibrio cholerae was the subject of this study. Delayed hypersensitivity reactions to skin test antigens were found to be markedly depressed in Bengali patients with cholera 24 h after fluid repletion. Skin test response rates were lower in children and in adults with the disease than in both normal adults and children or in adults with an equivalent degree of malnutrition. Patients with equal degrees of dehydration due to noncholera diarrhea were significantly less immunosuppressed. Concurrent depression of other manifestations of cell-mediated immunity was not found.


Clindamycin-associated enterocolitis in guinea pigs: evidence for a bacterial toxin.

Experimental enterocolitis was induced in guinea pigs by intraperitoneal injection of clindamycin. Specimens of feces were collected daily in phosphate-buffered saline (pH 7.0) and pooled every second day. The pooled samples were centrifuged to remove solids, and the supernatant was sterilized by membrane filtration. The sterile fecal supernatants were then dialyzed for 48 h against two 15-liter changes of phosphate-buffered saline and subsequently tested for toxicity in cultured monolayers of mouse adrenal cells. A filterable toxin(s) was found in the fecal supernatants on days 2, 4, and 6 postchallenge and not in pretreatment samples. The toxin(s) caused enterocolitis when administered orogastrically to healthy animals and altered the morphology of cultured mouse adrenal cells. The alteration of adrenal cell morphology was neutralized by specific antitoxin to Clostridium histolyticum.


Interactions between human granulocytes and Blastomyces dermatitidis.

We studied interactions in vitro between human granulocytes and the yeast-like form of Blastomyces dermatitidis, because granulocytes are prominent in the host response to systemic blastomycosis. In Boyden chamber assays, broth culture filtrates of B. dermatitidis contained levels of granulocyte chemotactic activity that were significantly higher than those present in similar culture filtrates of Histoplasma capsulatum and Cryptococcus neoformans, two fungi that characteristically do not elicit granulocytes in infected tissues. Microscopic study, including electron microscopy, demonstrated that granulocytes phagocytosed B. dermatitidis promptly and efficiently. Moreover, granulocytes emitted light (chemiluminescence) at a brisk rate during phagocytosis of B. dermatitidis, indicating activation of intracellular metabolic pathways. However, fungicidal assay showed that granulocytes (1:1 cell-yeast ratio, 10% serum) killed only 29% of the B. dermatitidis inoculum during 3 h of incubation. Taken together, these findings suggest that there is disparity between phagocytosis and intracellular killing of B. dermatitidis by human granulocytes, perhaps because of resistance of this fungus to granulocyte microbicidal mechanisms.


Histocompatibility-linked genetic control of susceptibility to age-dependent polioencephalomyelitis in mice.

Susceptibility to induction of immune polioencephalomyelitis (IPE) was found to be controlled by a gene that is closely linked to the H-2 complex. Whereas mice of the AKR (H-2k) strain were susceptible to IPE induction, H-2-congenic mice, AKR.H-2b (H-2b from C57BL/6) and AKR.M (H-2m), were resistant. However, susceptibility to IPE may be under additional control by a gene(s) outside of the H-2 region, since both C57BL/6 (H-2b) mice and congenic B6.H-2k mice (H-2k from AKR) were resistant to IPE induction. F1 hybrid mice derived from AKR (susceptible) and DBA/2 (resistant) mice were susceptible to IPE induction, indicating that susceptibility is dominant in at least one gene, but susceptibility developed at a later age in the hybrid mice than in AKR mice. B6.PL-Ly-2a Ly-3a/Cy, C57BR, C57L, PL, and RF strain mice were resistant to ipe induction. Thus, of the 12 inbred strains tested so far, only two (C58 and AKR) are susceptible to IPE.


Defective prostaglandin synthesis by C3H/HeJ mouse macrophages stimulated with endotoxin preparations.

Macrophages obtained from C3H/HeN mice produced significant amounts of prostaglandin E when exposed to phenol-extracted lipopolysaccharides (LPS), whereas macrophages from C3H/HeJ mice were unresponsive. The lipid A fraction from phenol-extracted LPS was an effective inducer or prostaglandin synthesis by macrophages from C3H/HeN mice. The polysaccharide portion of the LPS molecule had no effect. In contrast, the C3H/HeJ macrophages did not produce prostaglandin E in response to the lipid A moiety of phenol-extracted LPS. LPS prepared by butanol extraction stimulated the production of prostaglandin E by macrophages from both C3H/HeN and C3H/HeJ mice. The component of butanol-extracted LPS that stimulated the C3H/HeJ macrophages was shown to be a lipid A-associated protein. Further studies demonstrated a correlation between prostaglandin production by the macrophages of these two strains of mice in response to butanol- and phenol-extracted LPS and the lethal effects of the endotoxin preparations.


Large-scale production and partial purification of mouse immune interferon.

Large-scale production of high-titered (10(2.2) to 10(4) U/ml) immune interferon (type II) was carried out in roller cultures of mouse spleen cells by using the T-cell mitogen staphylococcal enterotoxin A. Precipitation of 90% of this interferon by 55 to 80% saturated ammonium sulfate resulted in a 20-fold concentration and a two- to sixfold purification. After application of this interferon to either bovine serum albumin (BSA)-Affi-Gel 10 or hydroxylapatite columns, 100% of the interferon activity was recovered. By BSA-Affi-Gel 10 chromatography, 7% of the recovered activity was not bound, 45% was eluted with pH gradient 5 to 7, and 48% was eluted with 1 M NaCl. The pH- and salt-eluted interferons from the BSA-Affi-Gel 10 column were purified 62- and 390-fold, respectively, when compared with the starting materials. Rechromatography of the pH- and salt-eluted interferon peaks from the BSA-Affi-Gel 10 column did not alter their elution patterns. Stepwise elution of interferon from the BSA-Affi-Gel 10 columns with buffers of various pH and salt contents also resulted in greater than 300-fold purification. Specific activities of up to 2 x 10(5) U of interferon per mg of protein were attained with either elution procedure from BSA-Affi-Gel 10 columns. By hydroxylapatite chromatography, 5% of the recovered activity was not bound, 20% was eluted with a salt gradient, and 75% was eluted with 30% glycerin. Purification was 107- and 16-fold, respectively, for the two fractions. Ultrogel AcA 34 chromatography of the interferon resulted in two peaks of activity, a major one with a molecular weight of approximately 40,000 and a minor peak of molecular weight 70,000 to 90,000. Thus, by different types of chromatography, immune interferon was found to be heterogeneous.


Delayed-type hypersensitivity against Semliki Forest virus in mice.

After intracutaneous immunization with purified inactivated Semliki Forest virus, a delayed-type hypersensitivity without detectable antibodies in serum was obtained in BALB/c mice. Low doses of antigen given intraperitoneally induced antibodies. Intracutaneous immunization with much higher doses induced no specific antibodies, but a footpad swelling was observed after challenge with homolgous antigen. Pretreatment with cyclophosphamide before immunization enhanced footpad swelling. Microscopic examination of footpads from sensitized mice at 24 h after challenge showed a mononuclear infiltrate. The delayed-type reaction could be transferred to syngenic mice with lymph node cells, but not with spleen cells or serum. The biphasic character of the delayed-type hypersensitivity is discussed.


Temperature-sensitive mutants of influenza A virus: evaluation of the A/Victoria/75-ts-1A2 temperature-sensitive recombinant virus in seronegative adult volunteers.

An influenza A virus recombinant bearing the surface antigens of the A/Victoria/3/75 (H3N2) strain and the two ts genes of the A/Udorn/72-ts-1A2 virus was evaluated for attenuation, antigenicity, and protective effect in 42 doubly seronegative adult volunteers (i.e., individuals who lacked detectable serum antibodies for the hemagglutinin and neuraminidase antigens). This recombinant, which had a 37 degrees C shutoff temperature for plaque formation and ts mutations on the genes thought to code for the P1 and P3 polymerase proteins, infected 90% of the volunteers. Of the volunteers, 5% developed mild coryza or rhinitis but other signs or symptoms were not observed, indicating that the A/Victoria/75-ts-1A2 recombinant was more attenuated than the A/Victoria/75-ts-1[E] recombinant. Vaccinees shed virus for a shorter interval and at a lower titer than did the A/Victoria/75-ts-1[E] vaccinees. Each ts-1A2 isolate retained the ts phenotype indicating that the recombinant was stable genetically in doubly seronegative adults. Finally, the ts-1A2 recombinant induced significant resistance to subsequent challenge with A/Victoria/75 wild-type virus.


Cold-adapted variants of influenza A virus: evaluation in adult seronegative volunteers of A/Scotland/840/74 and A/Victoria/3/75 cold-adapted recombinants derived from the cold-adapted A/Ann Arbor/6/60 strain.

Influenza A/Scotland/74 (H3N2) and A/Victoria/75 (H3N2) cold-adapted (ca) recombinant viruses, prepared by mating the A/Ann Arbor/6/60 (H2N2) ca donor virus and influenza A wild-type virus, were evaluated in adult seronegative volunteers (serum hemagglutination-inhibiting antibody titer, </=1:8) for level of attenuation, antigenicity, and genetic stability of the temperature-sensitive and ca phenotypes. At 10(7.0) to 10(7.5) 50% tissue culture infective doses the A/Scotland/74 and A/Victoria/75 ca recombinant viruses were clearly attenuated and antigenic. However, one of eight vaccinees infected with 10(7.5) 50% tissue culture infective doses of the A/Scotland/74 ca recombinant had a febrile reaction (39 degrees C). At a 10-fold higher dose (10(8.5) 50% tissue culture infective doses), 4 of 12 A/Scotland/74 vaccinees had a febrile and/or systemic reaction. Febrile reactions were not observed in volunteers who received the A/Victoria/75 ca recombinant virus, whereas 3 of the 12 vaccinees had mild upper respiratory tract symptoms, in one instance associated with mild systemic manifestations. Significantly, the serum hemagglutination- and neuraminidase-inhibiting antibody responses were comparable to those induced by wild-type virus. Both ca recombinant viruses were shed in low titer for a short period of time. Each isolate retained the temperature-sensitive phenotype. However, there was evidence of genetic instability of the ca marker in that 7 of 24 isolates exhibited some loss of the ca property, and one isolate completely lost the capacity to produce plaques at 25 degrees C. The retention of a low level of residual reactogenicity in the A/Scotland/74 ca recombinant suggests that acquisition of the ca and temperature-sensitive phenotypes by a ca recombinant virus may not always bring about a satisfactory level of attenuation for individuals lacking hemagglutinin immunity.


Cell-free and cell-bound antibody in nasal secretions from infants with respiratory syncytial virus infection.

Twenty-two infants under 9 months of age hospitalized with bronchiolitis or pneumonia due to respiratory syncytial virus (RSV) were serially sampled to determine the pattern of secretory antibody response. Using double labeling techniques, we found several types of immunoglobulin in secretions: cell-free antibody to RSV of the immunoglobulin A (IgA), immunoglobulin G (IgG), and immunoglobulin M (IgM) classes; and immunoglobulins of all three classes bound to RSV-infected cells shed from the nasal epithelium (presumably cell-bound antibody to RSV). IgA attached to RSV-infected epithelial cells was almost always detected in the first available nasal sample (day 1 or 2 of hospitalization). In contrast, cell-free anti-RSV IgA first appeared an average of 3.5 days later at a time when virus antigen was disappearing from the secretion. IgG and IgM attached to RSV-infected cells appeared more irregularly. The titer of cell-free anti-RSV IgM was often higher than that of IgA early in the illness and declined as the infection resolved. Cell-free anti-RSV IgG was usually present earlier than IgA and rose during convalescence.


Chemotactic deactivation of human neutrophils: possible relationship to stimulation of oxidative metabolism.

Neutrophils preexposed to high concentrations of activated complement or synthetic N-formyl methionyl peptides are inhibited in their subsequent spontaneous and chemotactic migratory responses. We have considered the possibility that a part of this nonspecific loss of migratory function may be attributable to the interaction of the leukocytes with reactive forms of oxygen deriving from the cytotaxin-induced burst of oxidative metabolic activity. For these studies we have assessed the effect of preexposure of neutrophils from patients with chronic granulomatous disease to cytotaxins on their subsequent migratory responses. We find that these responses are not altered by preexposure to either cytotaxin. Thus, there appears to be a functional relationship between deactivation and the ability of the normal neutrophil to undergo a cytotaxin-induced respiratory burst.


Tumor induction by Agrobacterium tumefaciens prevented in Vigna sinensis seedlings systemically infected by ribonucleic acid viruses.

Cowpea (Vigna sinensis) seedlings failed to develop tumors after being inoculated with crown gall bacteria (Agrobacterium tumefaciens) if, at times earlier than 1 day later, they were inoculated on the primary leaves with a cowpea mosaic virus that systemically infects them. Inoculation with buffer or with a virus that is restricted to a localized infection, or to which the cowpea is immune, did not interfere with the subsequent development of tumors. The virus infection did not appear to affect directly the titer of A. tumefaciens in the inoculation sites. Nor did mixing of virus particles with A. tumefaciens prevent subsequent appearance of tumors. The influence of virus infection extended across grafts (into tissue that is not susceptible to the virus) and there prevented tumor formation. The sap from infected plants, but not purified virus, decreased tumor formation on carrot disks. Systemic virus infection may induce in cowpeas a translocated substance that prevents tumor induction by A. tumefaciens.


Experimental sporotrichosis in Syrian hamsters.

Syrian hamsters were infected with Sporothrix schenckii by subcutaneous footpad inoculation. Two types of infection could be uniformly induced: a self-limited, lymphatic infection resembling the classical disease in humans, and a generalized nonfatal infection. An infecting dose of approximately 5,300 yeast cells produced the localized subcutaneous-lymphatic disease which was limited to a single limb. In contrast, a 1,000-fold increase in the inoculum temporarily overwhelmed the animals' defense mechanisms, producing a systemic infection involving the liver and spleen. These models were used to demonstrate the development of increased resistance to subsequent infection following either infection or active immunization with ribosomal fractions or trypsinized cell wall antigens of S. schenckii incorporated in Freund complete adjuvant. Agglutination titers were detectable in all animals that were either infected or immunized. In one group of infected animals, the titers persisted for at least 1 year after three booster doses of Formalin-killed S. schenckii. The ability to produce an infection in hamsters which closely resembles the disease seen in humans makes the animal a good model with which to study experimental sporotrichosis.


Route of infection, systemic host resistance, and integrity of ganglionic axons influence acute and latent herpes simplex virus infection of the superior cervical ganglion.

The character of acute and latent herpes simplex virus (HSV) infection of the superior cervical ganglion (SCG) in mice depended on the route by which the virus reached the ganglion, the level of systemic host resistance, and the integrity of postganglionic nerves. Prevention of ganglionic infection by postganglionic neurectomy carried out before intraocular (i.o.) virus challenge established the importance of the neural route in the development of SCG infection. However, hematogenous virus dissemination also led to SCG infection although with reduced frequency compared to that with i.o. inoculation. Enhanced host systemic antiviral resistance had two divergent effects on ganglionic infection depending on the dose and timing of virus inoculation. Thus, both acute and latent ganglionic infections were concomitantly reduced when resistant C57B1/6 mice were challenged with low doses of virus or when less resistant BALB/c mice were actively immunized 1 week before virus challenge. On the other hand, when resistant mice were challenged with high doses of virus or when either active or passive (antibody) immunization was delayed long enough to assure viral access to the ganglion, intraganglionic viral replication during the acute phase of infection was reduced, but the prevalence of subsequent latent infection was either unaffected or actually enhanced. Postganglionic neurectomy, performed after virus had reached the ganglion, altered the course of SCG infection in a direction opposite that of immunization, augmenting the acute phase of viral replication while reducing latency. In athymic nude mice and mice immunosuppressed with cyclophosphamide, intraganglionic viral replication was prolonged. These results emphasize that host factors both extrinsic and intrinsic to the SCG modify the course of ganglionic infection.


Host defenses in murine malaria: evaluation of the mechanisms of immunity to Plasmodium yoelii infection.

The immune response of random-bred mice to infection with a relatively avirulent strain of Plasmodium yoelii was measured in terms of parasitemia, splenomegaly, immediate and delayed hypersensitivity to a P. yoelii antigen preparation, resistance to challenge with a virulent variant of P. yoelii, and nonspecific resistance to L. monocytogenes. Avirulent P. yoelii produced a self limiting infection which resolved in 21 days. Peak parasitemia and splenomegaly were observed at 14 days, and infected mice were resistant to challenge with virulent P. yoelii from 7 days through at least 126 days. Mice infected with avirulent P. yoelii developed humoral immunity as judged by immediate hypersensitivity reactions and the capacity of their serum to passively protect normal mice against virulent P. yoelii. At no time did mice infected with the avirulent P. yoelii display evidence of cell-mediated immunity, as expressed by delayed-type hypersensitivity and increased resistance to L. monocytogenes. In fact, at the height of avirulent P. yoelii infection there was decreased resistance to L. monocytogenes in both liver and spleen, and the macrophages of the undisturbed peritoneal cavity were similarly defective. It was concluded that the defense mechanism of mice against P. yoelii is mediated by humoral factors in the absence of demonstrable cell-mediated immunity.


Human cell-mediated immune responses to chlamydial antigens.

A reproducible method was developed to determine the ability of chlamydial antigens to stimulate lymphocytes from volunteers. In tests repeated 4 to 14 times, the cells from a given volunteer gave a relatively narrow range of responses, but there were great differences in the mean response of different volunteers. In the entire group of 52 volunteers, lymphocyte stimulation was significantly associated with the presence of antibody, but in a given individual results of one test did not aid in predicting the results of the other. A majority of persons with either antichlamydial antibody or elevated lymphocyte stimulation, or both, did not have a history of signs or symptoms within a spectrum of chlamydial diseases. This may reflect the great frequency of asymptomatic infection with these organisms. The lymphocytes of some individuals were stimulated to a significantly greater degree by antigens of one chlamydial species (Chlamydia trachomatis or C. psittaci) than by the other. These and other cell-mediated reactions in human chlamydial infections, and their possible medical significance, are under continued study.


Preparation of glucosyltransferase from Streptococcus mutans by elution from water-insoluble polysaccharide with a dissociating solvent.

Glucosyltransferase (EC 2.4.1.5) was obtained by dissociation from water-insoluble polysaccharide in the presence of 6 M guanidine-hydrochloride. Water-insoluble polysaccharide was synthesized by cell-free culture supernatants from Streptococcus mutans strain 6715. Gel filtration of the glucosyltransferase on a column of 8% agarose in phosphate buffer, followed by filtration on a column of 4% cross-linked agarose in 6 M guanidine-hydrochloride, gave a 23-fold enrichment of the enzyme. The enriched glucosyltransferase preparation contained 22% carbohydrate and eluted at a position corresponding to a molecular weight of 422,000. Polyacrylamide gel (5%) electrophoresis of this preparation revealed two regions which stained for protein, formed water-insoluble polysaccharide in the presence of sucrose, and precipitated with antisera directed to crude glucosyltransferase preparations. The guanidine-eluted enzyme could be primed by 5 X 10(-5) M dextran T10 (molecular weight, 10,000). High-molecular-weight glucan and a possible glucan-binding protein were also obtained after the final gel filtration step (4% cross-linked agarose) in addition to glucosyltransferase.


Common antigens among systemic disease fungi analyzed by two-dimensional immunoelectrophoresis.

The interpretation of immunological results in systemic mycoses has been complicated by cross-reactions among specimens from patients with blastomycosis, coccidioidomycosis, and histoplasmosis. The fungal preparations used in these tests evidently contained one or more antigens in common. Two-dimensional immunoelectrophoresis techniques were used to determine the number of antigens contained in several soluble extracts from Blastomyces dermatitidis and Histoplasma capsulatum that were common with those demonstrable in a coccidiodin-anticoccidioidin reference system. A total of 12 and 10 common antigens were found in preparations from B. dermatitidis and H. capsulatum, respectively. In addition, the crude preparations from each species of fungus exhibited some qualitative and quantitative differences in composition of antigens. Use of two-dimensional immunoelectrophoresis for standardization of fungus extracts, for monitoring separation of species-specific antigens, and for testing humoral antibody response should add further refinement to correlations with clinical disease.


Cryptic peptidoglycan and the antiphagocytic effect of the Staphylococcus aureus capsule: model for the antiphagocytic effect of bacterial cell surface polymers.

The antiphagocytic effect of the Staphylococcus aureus capsule is known to be related to its ability to interfere with opsonization by normal human serum. In this study, evidence is presented with isolated cell surface components which indicates that the capsule hinders opsonization by masking cell wall peptidoglycan. In contrast to intact, encapsulated S. aureus M cells, peptidoglycan particles isolated from the organism were efficiently opsonized by normal human serum and phagocytized by human polymorphonuclear leukocytes. Cell wall particles retaining capsular material were opsonized less efficiently than peptidoglycan. Studies comparing the opsonic capacities of normal, C2-deficient, and heat-inactivated sera led to the conclusion that both the classical and the alternative complement pathways contribute to the opsonization of peptidoglycan in normal human serum. It appears that the capsule interferes with opsonization via both of these complement pathways. Serum from rabbits immunized with S. aureus M had significant heat-stable opsonic activity for the intact organism and cell walls retaining capsular material, but not for peptidoglycan. A general model is proposed to explain how antiphagocytic cell surface polymers may inhibit bacterial opsonization and thereby impede natural immunity.


The nitroblue-tetrazolium test in granulocytes of the cerebrospinal fluid--methodological problems.

It was originally our intention to study the suitability of the nitroblue-tetrazolium (NBT) test for differentiating the causes of granulocytic pleocytosis of the cerebrospinal fluid (CSF). However, several methodological problems were encountered and required extensive preliminary studies. The test was initially performed on 51 CSF samples from 37 patients using the method described by Park et al. in 1968. Although bacterial meningitis was demonstrated in 19 patients, the NBT test resulted in more than 11% NBT positive granulocytes in only four cases. No NBT positive cells were found in 24 of the samples tested. These rather poor results are not surprising if one considers that NBT is only taken up by granulocytes as macrocomplex containing heparin or fibrinogen. CSF does not contain measurable quantities of fibrinogen--either under normal conditions or in cases of viral or bacterial meningitis. A comparative study of the NBT test with and without the addition of heparin was performed using CSF samples from six patients with proven bacterial meningitis. Without heparin there were less than 11% NBT positive granulocytes in five cases, while all samples demonstrated more than 11% positive cells after addition of heparin. Best results were achieved with 70 IU of heparin in each sample. Preparation and evaluation were facilitated by use of 0.1% NBT solution and concentration of the CSF cells in a sedimentation chamber.


[Current chemotherapy of infections caused by Pseudomonas aeruginosa (author's transl)].

In addition to carbenicillin, the newer beta-lactam antibiotics such as ticarcillin and azlocillin are now available for the chemotherapy of Pseudomonas aeruginosa infections. We investigated the in vitro effect of these antibiotics on 233 isolates from clinical material. We were particularly careful, when choosing the experimental material, to exclude copies of the same individual strain, and we achieved this by combining various epidemiological typing procedures. A comparison of carbenicillin, ticarcillin and azlocillin according to the concentrations at which half of the 233 strains were inhibited showed the ticarcillin values to be higher than those of azlocillin by a factor of 2.1, and carbenicillin values to be higher than those of azlocillin by a factor of 4.9. Individual strains also occurred in which the inhibitory concentration for azlocillin was higher than that of carbenicillin (5 strains) or ticarcillin (11 strains). In 17 out of the 233 isolates no therapeutic success would have been within reach even with the newer beta-lactam antibiotics. The use of ticarcillin and azlocillin permits an extension of the indications for therapy with beta-lactam antibiotics in P. aeruginosa infections, from hitherto 76%, to 90% of the cases. If one includes the aminoglycosides gentamycin, tobramycin, sisomycin and amikacin in the therapeutic armoury, then the proportion of in vitro sensitive strains of P. aeruginosa in the material submitted for examination rises to 98%.


Pharmacokinetics of intramuscular carbenicillin.

A 500 mg intramuscular dose of carbenicillin produced peak levels averaging 147 microgram/ml after three hours in the first day full-term newborn infants, and 172 microgram/ml after one to two hours in infants five days of age. The fall of blood levels thereafter was delayed and serum half-life averaged 4.2 and 2.2 hours in both groups, almost four times and twice as long, respectively, as that reported in adults. Absorption from the injected site was also delayed in young newborns, as was shown by the delayed serum peak and small estimated absorption rate constant. This must be taken into consideration if the intramuscular route is chosen in young newborn infants. On the basis of serum half-life, an administration interval of 12 hours was recommended for newborns younger than four days, and eight hours for those five days of age or more.


Lack of penetration of cephacetril into the cerebro-spinal fluid of patients without meningitis.

Six neurosurgical patients were given 2-4 g cephacetril intravenously at six hourly intervals. Serum and cerebrospinal fluid (CSF) concentrations were determined. Cephacetril activity was not detectable in the CSF when the blood-CSF barrier was intact. In two patients with increased CSF cell counts cephacetril was found in the CSF in low concentrations (0.1-0.8 mcg/ml). Our investigations show that cephacetril does not penetrate into the CSF unless there is an inflammation or other impairment of the blood-CSF barrier. Thus we could not confirm the results of some authors who found cephacetril in therapeutically effective concentrations in the CSF of healthy persons.


A defect in neutrophil motility in two siblings with recurrent infections and a remarkable family history.

Two siblings with recurrent infections were found to have impaired neutrophil motility. The same association of infections (otitis media, bronchitis, chronic diarrhoea) has caused seven fatalities in the paternal side of the family, suggesting genetic implications.


Delayed hypersensitivity to soluble antigens in mice. II. Analysis in vitro.

Delayed hypersensitivity to a soluble protein has been analyzed in vitro with lymph node cells from mice. Antigen-dependent cytotoxicity against innocent bystander cells, and incorporation of tritiated thymidine have been used to monitor the response to a subcutaneous tail injection of egg albumin (OVA) in complete Freund's adjuvant. Sensitized lymph node cells from mice with positive 24 h ear tests killed A9 cells in the presence of antigen. This antigen-specific, T cell-mediated cytotoxicity was observed within 4 days of sensitization and still apparent at 7 months. Reactivity to OVA, assessed as an increase in DNA synthesis in the presence of antigen, was also T cell dependent and exhibited similar sensitization kinetics and dose requirements. Both microtechniques require as few as 10(5) lymphocytes and are readily adaptable to the study of delayed hypersensitivity to any soluble protein in mice.


Pregnancy-associated a2-glycoprotein in rheumatoid arthritis.

Pregnancy-associated alpha2-glycoprotein (PAG) was found to be increased in sera and synovial fluids of patients suffering from rheumatoid arthritis tested in the single radial immunodiffusion. The PAG levels in rheumatoid sera correlated well with the activity of arthritis in the patients. The purified PAG inhibited clearly the 3H-thymidine uptake of normal lymphocytes stimulated with phytohemagglutinin and also the spontaneous attachment of sheep erythrocytes to normal lymphocytes. 17 of 19 rheumatoid plasmas inhibited the 3H-thymidine uptake of normal lymphocytes and intensity of their inhibitory effects correlated with serum PAG levels.


Humoral immunoreactivity in chronic active hepatitis: relation to HLA antigens.

43 patients with chronic active hepatitis (CAH) exhibited significantly higher levels of antibodies against measles and polio type 2 und 3 when compared to 43 age- and sex-matched healthy controls. No significant differences were found for antibodies against polio type 1, rubella, herpes simplex, mumps virus, and tetanus. There was no correlation between antibody levels and the presence of HLA-B8 and/or HLA-B12. The antibody response to vaccination with polio vaccine (killed) and tetanus toxoid was not higher in CAH patients carrying HLA-B8 and/or HLA-B12 than in patients without these antigens. The results indicate that CAH is associated with an increased immunoreactivity, which is, however, not linked to HLA-B8 and/or HLA-B12.


A solid-phase radioimmunoassay to detect anti-tuberculin antibodies in the guinea pig.

Despite the importance of the guinea pig as a model test animal for studying the humoral immune response in comparison to the development of cellular immunity after BCG or tuberculosis infection, no precise and sensitive method for the determination of such antibodies is available. We developed an indirect solid-phase radioimmunoassay on microtiter plates for this purpose. All assay conditions have been optimized such as selection of suitable plastic material, use of carrier substances, concentrations and amount of the antigen as well as the serum and second antibody, buffer, incubation times and all steps of the procedures for coating, washing, and measuring. The assay gives a linear relationship between antibody concentration in the serum to be tested and binding of the radiolabelled second antibody. A comparison with four conventional antibody titration methods reveals a 128- to 512-fold higher sensitivity of the RIA. Favorable aspects of this RIA as opposed to the methods hitherto used are discussed.


Lentil-lectin affinity chromatography of surface glycoproteins and the receptor for IgE from rat basophilic leukemia cells.

Surface proteins and glycoproteins of RBL cells were labelled enzymatically with 125I and then solubilized with Nonidet P-40. Analysis by polyacrylamide-gel electrophoresis in SDS on 10% gel revealed 10 distinctive peaks ranging in molecular weights from 17,000 to 200,000 daltons. Mainly components of higher molecular weights were bound by lentil-lectin Sepharose and could be eluted with alpha-methyl mannoside. The receptor for IgE was clearly shown to bind to the lentil-lectin. A second cell surface component which previously had been shown to bind to IgE-Sepharose as well, was found to bind only slightly to lentil-lectin. Thus, it can be concluded that the receptor for IgE is a glycoprotein with mannose and/or N-acetylglucosamine in the carbohydrate moiety(s).


Comparison of the atopic background between allergic and non-allergic beekeepers.

A study was carried out on beekeepers and their families. 34 subjects with a history of bee sting allergy and also a positive radioallergosorbent test (RAST) to honey bee venom were compared with 47 subjects with neither. 16 subjects in the bee allergy group (47%) had a history of atopic diseases as compared to 6 (13%) in the control group (p less than 0.01). 15 in the bee allergy group (44%) had at least one positive RAST to three common inhalant allergens as compared to 5 (11%) in the control group (p less than 0.01). These results suggest that, although bee sting allergy frequently affects non-atopic subjects, sensitization among beekeepers and their families occurs more readily in atopics. It is notable that a large proportion of patients injected with foreign proteins (the control group) either never develop venom-specific IgE antibodies or have suppressed their production.


On the latency period of sensitization: inhibition of passive cutaneous anaphylaxis by a factor in normal rat serum.

Addition of normal rat serum to mouse or rat IgE antibody blocks the passive cutaneous sensitization of rats with either IgE. The inhibition can be obtained with isologous and autologous rat serum. The blocking factor was purified and characterized as a molecule with a MW of 69,000 daltons, a sedimentation rate of 4.5 s and an isoelectric point of pH 4.7. The factor blocks the PCA reaction also at previously sensitized skin sites and seems to act on the secretory phase of mediator release.


Standardization of radioallergosorbent test (RAST) and allergen extracts in terms of the WHO standard for IgE.

An 131I-labelled IgE myeloma, standardized against the WHO reference serum for IgE was bound in varying concentrations to discs bearing anti-human light chain. These were used to construct a calibration curve for the subsequent binding of 125I-labelled anti-IgE, thus relating 125I counts to WHO IgE units. The calibrated anti-IgE could then be used to measure the actual amount of specific IgE bound to an allergen disc in the radioallergosorbent test. Using this standardized system, soluble allergen extracts could then be assayed by their ability to inhibit the binding of a defined amount of specific IgE, thereby enabling their activity to be expressed in WHO IgE units.


Effect of cobra venom factor on the response of Wistar-Furth rats to the Gross-virus-induced (C58NT)D lymphoma.

Wistar-Furth rats were injected with the anticomplementary cobra venom factor at the time of challenge with the Gross-virus-induced (C58NT)D lymphoma. Complement depletion and/or activation of the complement system in these animals led to the following findings: Multiplication of the tumor cells in vivo was accelerated and mortality rates were enhanced. In addition, serum levels of the immunoglobulins which mediate antibody-dependent cellular cytotoxicity were depressed.


Relation between total serum IgE levels and IgE antibody production in rats.

Five different rat strains were immunized with 100 microgram ovalbumin and 1 mg Al (OH)3. A good correlation was found between the total serum IgE level of a rat strain before immunization and the IgE antibody production. Good IgE antibody-producing strains had high total serum IgE levels and vice versa. A discordance between the evolution of the total serum IgE level after immunization and the changes in IgE antibody level was found in all strains.


Half-body x-ray suppresses the acquisition of delayed-type hypersensitivity in the mouse.

800 R half-body x-ray suppressed the acquisition of allergic contact dermatitis in mice (1) if the x-ray field included the lymph nodes regional to the sensitization site, and (2) when x-ray was given during, but not immediately before, induction of the sensitivity.


Skin manifestations and serum IgE levels in levamisole-treated rheumatoid arthritis patients.

Skin manifestations may occur as adverse effects of immunostimulant therapy with Levamisole. They are generally regarded as drug-induced hypersensitivity reactions. Our observations revealed that, in Levamisole-treated rheumatoid arthritis patients, urticaria was accompanied by an elevation of serum IgE, but other types of skin reactions were not. Moreover, elevation of IgE occurred without any skin reactions in some cases. It is suggested that Levamisole - stimulating T lymphocytes - may also stimulate IgE-mediated atopic responses.


Pricing, demanders, and the supply of health care.

This paper is divided into three substantive sections. In the first section the conventional neoclassical paradigm is augmented by consideration of the agency relationship in which the physician is considered not only as the agent who controls the supply of health care, but also as the decision maker who articulates demand because patients forego this role and rely on "expert" advice. The next section is concerned with the effects of pricing on consumer demand and draws on the available empirical evidence to present estimates of price elasticity, cross elasticity, and other characteristics of the choice process. This analysis is completed by integrating the agency relationship into the discussion and arguing that if the policy objectives are expenditure containment and greater efficiency in resource utilization, the price mechanism should be used to affect the behavior of the primary demander and the supplier: the physician. In the final section the implications of this analysis are discussed in the contexts of two competing perspectives: the liberal market perspective and the collective "needology" perspective, and an attempt is made to distinguish some of the characteristics of the two views of the world.


Third World experiences in health planning.

In response to an invitation by the American Public Health Association, the author discusses his experiences in health work with particular reference to the Third World. These cover a period of four decades of activities in many countries, the discussion being primarily directed toward the North American audience attending the panel organized by the International Health Committee at the 104th Annual Meeting of the Association in Miami Beach in October 1976. First the paper deals with the legacy of broad social teaching resulting from the years of international collaboration from the time of Franklin D. Roosevelt to that of Richard Nixon. Public health problems, whether new or old, are essentially social in character and can only be solved in terms of social policy. Attention is directed to the current mistake of placing the emphasis on individual behavior, divorced from its social base, in the work of health professionals servings in Third World countries. The weakness of national average values and the consequent need of measuring the differentials between social groups and classes are widely illustrated. Finally, positive and negative lessons learned by experimenting with health technology consistent with the expected development of countries are examined as a basis for a genuinely emancipatory approach to the health problems in the Third World.


Prevention: rhetoric and reality.

No single strategy is capable of preventing untimely deaths and disabilities. In the United States in particular, the contribution of medical care is limited by inadequate provision of services (particularly to the poor), inappropriate training of physicians, and unnecessary costs. Lack of knowledge about disease universally limits medicine's effectiveness. Among nonmedical strategies, campaigns for life-style change are most likely to succeed in those with the lightest burden of illness. Efforts to increase individual responsibility might well reduce health expenditures, but at the same time the disparity in health between rich and poor will increase. Restrictions on the use of harmful substances and on the manufacture of toxic or hazardous products, along with humanization of the work process itself, would reduce deaths from cancer, heart disease, and violence; however, resistance to these changes is, and will remain, great. The socioeconomic and environmental changes that are necessary to afford each citizen an equal opportunity for optimal health will be adopted slowly, if at all. Considering the limited acceptability of these nonmedical strategies, the prospects for prevention are less than what has been promised. Despite its perfections, medical care can contribute to the prevention of early death and disability. Attempts to disparage it could delay the adoption of the changes needed to improve its effectiveness.


Job alienation and well-being.

This paper analyzes the relationships between alienation conceived as monotonous, repetitive work and alienation as job-worker incongruence to a variety of measures of worker as well-being among a population of workers from Victoria, British Columbia. The data show weak relationships between work perceived as monotonous and general psychological and physical well-being and between alienation as job-worker incongruence and health. While weak in variance-explained terms, the relationships show the predicted patterns are robust and are independent of a large number of control variables. Percentage differences in well-being between the alienated and nonalienated workers are fairly substantial. Some societal implications of the findings are discussed.


Human rights, health, and capital accumulation in the Third World.

This article examines the relationship between human rights and the pattern of capital accumulation in the Third World. The repressive authoritarian State increasingly constitutes the means for enforcing the intensive exploitation of labor in Third World industrial enclaves and commercial agriculture. While the development of center capitalism has evolved toward "the Welfare State" and a framework of liberal sociodemocracy, the "peripheral State" is generally characterized by nondemocratic forms of government. This bipolarity in the state structure between center and periphery is functionally related to the international division of labor and the unity of production and circulation on a world level. The programs and policies of the center Welfare State (health, education, social security, etc.) constitute an input of "human capital" into the high-technology center labor process. Moreover, welfare programs in center countries activate the process of circulation by sustaining high levels of consumer demand. In underdeveloped countries, the underlying  vacuum in the social sectors and the important allocations to military expenditure support the requirements of the peripheral labor process. Programs in health in the center and periphery are related to the bipolarity (qualification/dequalification) in the international division of labor. The social and economic functions of health programs are intimately related to the organic structure of the State and the mechanics whereby the State allocates its financial surplus in support of both capitalist production and circulation.


Demographic growth and health needs in Latin America.

The net effect of the rapid population growth in Latin American countries is an increase in the needs for health services. Nevertheless, the demographic factor is not the only nor the most important consideration in determining how to satisfy these demands in the region. The main contradiction lies between the magnitude of needs for services generated by the adverse living conditions prevalling among the majority of the population, together with a restricted supply of health services, the availability of which varies according to social class. The problem of the increasing demand for medical care, generated by the rapid population growth, should be recognized as originating in the socioeconomic structural conditions prevailing in Latin American countries today which determine, simultaneously, low health levels, deficiencies in the provision of health services, and rapid population growth.


Professional ideology and the health care system in Chile.

Doctors' ideology and the role the medical profession plays in the organization and development of health services of a particular country are subjects of scientific interest. This article examines the case of Chile, where doctors centralized administrative and professional power over the type of health services provided to the population. Changes in this ideology throughout the century have been reflected in changes in the system of medical care. A shift can be traced from a professional ideology emphasizing socialized medical services during the first half of the century toward the current position favoring the abolition of free medical care and a return of "fee per service" medicine. The possible explanation for these changes is that medical ideology has altered through the combined influence of the class position of physicians and the development of the profession itself. Increasing polarization of the political forces in Chile led doctors to act according to their class affiliation, regardless of their original professional positions. Some historical facts are provided at each stage of analysis as a way of clarifying the conflicting influences upon the doctors' position.


Lidocaine plasma concentrations during and after endoscopic procedures.

Plasma lidocaine concentrations were intermittently measured in 8 upper gastrointestinal endoscopy and 12 bronchoscopy patients. The highest individual concentration was 0.98 microgram/ml in the upper gastrointestinal endoscopy patients and 3.79 microgram/ml in the bronchoscopy patients. Highest concentrations were reached at 15 minutes in the gastrointestinal endoscopy patients and at 30 or 60 minutes in the bronchoscopy patients. Thus, since lidocaine does not produce toxic effects at concentrations inferior to 6 microgram/ml, doses of this topical anaesthetic up to 16 mg/kg can be safely given during endoscopic procedures to patients with normal hepatic and cardiovascular functions. However, patients with liver metastases should be considered at high risk even if their liver function tests are normal. Patients at high risk of developing lidocaine toxicity should receive lower doses and be closely watched for at least 60 minutes after the end of the procedure.


Pharmacokinetics of l-carnitine in man following intravenous infusion of dl-carnitine.

The pharmacokinetics of l-carnitine were studied in adult male patients following intravenous infusion of dl-carnitine hydrochloride-L-Carnitine appears to distribute into a rapidly perfused and then a more slowly perfused body space following administration. The overall apparent distribution volume Vd (ss) is consistent with extracellular body water. Serum levels of l-carnitine, from two different dose levels were adequately described by a two-compartment model. 80% of the administered dose was recovered in 24 hour post dose urine.


Effect of adrenergic receptor blockade on cortisol and GH response to insulin-induced hypoglycemia in man.

The effect of several drugs presumably influencing central catecholaminergic receptors on plasma cortisol and GH response to insulin-induced hypoglycemia was studied in healthy adult males. The intravenous infusion of alpha-adrenergic blocking agents tolazoline or phentolamine supressed plasma cortisol and GH response to insulin-induced hypoglycemia. After an infusion of beta-adrenergic antagonist propranolol both hypoglycemia and rise in plasma cortisol and GH were prolonged. Finally, the administration of dopaminergic blocker pimozide failed to affect the plasma cortisol response, but slightly suppressed the enhancement of GH release during hypoglycemia. Caution is recommended before making suggestions about neuroendocrine regulations from the data obtained after systemic administration of drugs. Nevertheless, it may be concluded that the hypothesis on the inhibitory role of the central alpha-adrenergic system on ACTH secretion suggested in rats and dogs was not confirmed by our results obtained in man.


Antihypertensive effect and plasma levels of labetalol. A comparison with propranolol and dihydrallazine.

Seventeen outpatients suffering from essential hypertension were treated in a double-blind cross-over study with labetalol or with a combination of propranolol and dihydrallazine. The doses were increased depending on the response during the six week treatment periods. Both treatments reduced the blood pressure significantly as compared to the placebo, and the combination more than labetalol with the doses used, apparently because of a higher degree of the beta-blockade. Positive linear correlations were found between the dose of labetalol and the concentration in plasma as well as the concentration of labetalol in plasma and the decrease of standing blood pressures.


Placental transfer of digoxin (beta-methyl-digoxin) in man.

The dynamics of placental transfer of digoxin (beta-methyl-digoxin) has been studied in 20 pregnant women. The drug was administered in a single dose, intravenously, during labor. The samples drawn within the first hour after administration showed significantly lower fetal serum digoxin concentrations (SDC) than in the mother. No significant difference appeared in specimens collected after the first hour.


Cephapirin concentrations in prostatic and seminal vesicle tissues.

A 15 min i.v. infusion supplying either 1 or 2 g of sodium cephapirin was administered preoperatively to patients about to undergo either retropubic or transvesical prostatectomy. Tissue samples were obtained 30 min after the end of the infusion. Blood samples were obtained immediately before the start of the infusion and 30 min after the end of it. Cephapirin levels in the blood averaged 25.9 +/- 4.0 microgram/ml after the 1 g dose and 47.5 +/- 5.6 microgram/ml after the 2 g dose. Drug levels in prostatic tissue averaged 24.5 +/- 7.1 and 25.8 +/- 4.4 microgram/g for the 1 and 2 g doses, respectively. Levels in tissue taken from the seminal vesicle, often the focus of infection in bacterial prostatitis, averaged 12.5 +/- 2.3 and 44.8 +/- 14.9 microgram/g for the 1 and 2 g doses, respectively. These results suggest that bactericidal levels of cephapirin against sensitive organisms can readily be achieved in the prostate and seminal vesicles.


Pharmacokinetics of carbamazepine in the neonate and in the child.

1. Pharmacokinetic of carbamazepine were made in 7 new-borns and in 5 children. They were hospitalized for epilepsy and were receiving drugs such as phenobarbital alone or in association with other antiepileptic drugs, but not with carbamazepine. 2. The drug was given by oral route with a mean dose of 17.2 mg.kg-1. 3. The determination of carbamazepine concentration in serum was made by gas liquid chromatography on a 50 microliter sample. 4. A one compartment body model was used to determine the pharmacokinetic constants with first order rate constants for absorption and elimination. 5. Absorption was generally delayed by about half an hour, the maximum concentrations ranging from 3.14 to 10 microgram.ml-1 at 2 and 9 hr after administration. The mean half-life for absorption was 1.42 +/- 0.34 hr. The mean half-life for elimination was 8.76 +/- 0.85 hr. The half-life for elimination was much shorter than those already described even in multiple dosing epileptic adult patients. The pharmacokinetic parameters were used to predict blood levels in chronic treatment in 3 children. The predicted steady state concentrations disagreed with the concentrations measured.


Nummular eczema of statis origin. The backbone of a morphologic pattern of diverse etiology.

A total of 113 patients with nummular eruptions of unknown origin were collected over a period of 8 years. This number was reduced to 82 patients by exclusion of known clinical entities which became unmasked during the study. All 82 patients were found to have varicose veins and/or edema of the lower extremities. Autoeczematization may be the common denominator.


Immunologic and gastrointestinal abnormalities in dermatitis herpetiformis.

Eighty patients with untreated dermatitis herpetiformis (DH) were studied. The final diagnosis was made after completion of immunofluorescence (IF) studies of the skin, iodine provocation tests and gastrointestinal investigations. IgA deposits in the skin and the HLA-DRw3 B-cell alloantigen can be demonstrated in nearly all cases of DH. Positive iodine provocation test and gastrointestinal abnormalities are present in about 78% of the cases. For diagnostic purposes tissue typing and intestinal biopsy is of value in addition to IF studies of the skin.


Zosteriform inflammatory metastatic carcinoma.

A 57-year-old man presented with chest wall lesions and swelling of his left arm. The rapid onset of vesicular lesions in a dermatomal distribution resulted in an initial diagnosis of herpes zoster. Cutaneous biopsy revealed adenocarcinoma and further evaluation revealed a primary source of pulmonary adenocarcinoma. Lymphatic spread of tumor cells is the most likely source of the zosteriform skin lesions, but other possibilities are discussed.


Subcutaneous chromomycotic abscesses caused by Phialophora gougerotii.

Subcutaneous chromomycotic absesses due to phialophora gougerotii in a 50-year-old woman were characterized by subcutaneous abscesses adhering together through fistulous tracts, rupturing and leaving black crusts over multiple sinuses mimicking mycetoma. Reinfection occurred in contaminated excised lesions. Intradermal inoculation in different anemic patients caused abscesses in 14 days and healed without therapy in eight weeks. No spreading to other organs was found after intraperitoneal inoculation of mice for eight weeks.


Enzymatic dtermination of pyridoxal 5'-phosphate with gamma-cyanoaminobutyric acid aposynthase.

A rapid alternative method is presented for the determination of pyridoxal 5'-phosphate (pyridoxal-P). The method involves the colorimetric analysis of thiocyanate liberated from S-cyanohomocysteine (Hcy (CN)) in the presence of cyanide when catalyzed by the pyridoxal-P dependent enzyme, gamma-cyano-alpha-aminobutyric acid (gamma-CNabu)-synthase (Hcy (CN) thiocyano-lyase [adding CN]). The rate of formation of thiocyanate is determined by the increase in absorbance at 470 nm on treatment of the enzymatic reaction mixture with FeCl3.


Side reactions in peptide synthesis. X. Prevention of O-acylation during coupling with active esters.

Acylation of the hydroxyl groups in the side chains of serine, threonine or tyrosine occurs in coupling with active esters. This side reaction, which is quite pronounced in histidine-containing peptides, can be prevented with additives. From a series of compounds tested, 2,4-dinitrophenol and pentachlorophenol were the most effective.


Solid phase synthesis without repetitive acidolysis. Preparation of leucyl-alanyl-glycyl-valine using 9-fluorenylmethyloxycarbonylamino acids.

The utility of repetitive nonhydrolytic base cleavage of alpha-amino protective groups in solid phase peptide synthesis is shown by a preparation of the model tetrapeptide leucyl-alanyl-glycyl-valine on a p-benzyloxybenzyl ester polystyrene--1% divinylbenzene resin support. Nalpha-9-Fluorenylmethyloxycarbonyl (Fmoc: Carpino & Han, 1970, 1972) amino acids were coupled by the symmetrical anhydride procedure, followed by Fmoc group cleavage using 50% piperidine in methylene chloride. Quantitative removal of the Fmoc-tetrapeptide from the solid support was effected by treatment with 55% trifluoroacetic acid in methylene chloride. Homogeneous free tetrapeptide was obtained in 87% overall yield. The procedure is proposed to offer advantages over present solid phase methods which use acidolysis for repetitive alpha-amino group deblocking.


[21-arginine-A] insulin: a biologically active analog.

An analog of sheep insulin which differs from the parent molecule in that the C-terminal amino acid residue of the A chain, asparagine, is replaced by arginine, has been synthesized and isolated in highly purified form. The [Arg21] A chain of sheep insulin was synthesized by the fragment condensation approach and isolated as the  S-sulfonated derivative. Conversion of the latter into the sulfhydryl form and interaction with the S-sulfonated B chain of bovine (sheep) insulin yielded [Arg21-A] sheep insulin, which was purified by chromatography on a carboxymethylcellulose column with an exponential sodium chloride gradient. The [Arg21-A] sheep insulin shows potencies of 10.5--12.5 IU/mg when assayed by the mouse convulsion method and 8.6 IU/mg by the radioimmunoassay method (cf. 23--25 IU/mg for the natural hormone). It has been suggested that in the insulin molecule the A21 asparagine participates in salt bridge- and hydrogen bond-forming interactions which are critical in the biological activity of the hormone. Although the [Arg21-A] analog still retains these interactions, it is only ca. 50% as active as the natural hormone. It is speculated that other factors than the above mentioned interactions come into play, which involve the side chain of the A21 amino acid residue and affect the biological activity of the hormone.


The purification of peptides by partition chromatography based on a hydrophobicity scale.

A study of the efficiency of partition chromatography for the purification of peptides as a function of structure has been undertaken. A series of 19 omission analogs of camel beta-endorphin and of some of its partial sequences have been synthesized with each analog missing only a single amino acid. Their chromatographic properties have been examined with use of the Martin hypothesis and the RM concept, and a hydrophobicity scale for the amino acid side chains was obtained. To a first approximation a correlation with the Tanford hydrophobicity scale for amino acids was found. A decrease in hydrophobicity has been observed with increasing chain length and is discussed in terms of column efficiencies required for the purification of synthetic peptides.


Rabbit lutropin: preparation, characterization of the hormone, its subunits and radioimmunoassay.

The purification of rabbit lutropin is described. A product with a potency of 1.53 X NIH-LH-Sl was obtained as assayed by the ovarian ascorbic acid depletion assay. In a homologous radioimmunoassay, which is described, rabbit lutropin has a potency 4.83 X NIH-LH-Sl. In a radioligand assay, utilizing labeled ovine lutropin as the trace, the relative potency was 0.47 X NIH-LH-Sl measured by 50% inhibition comparison since rabbit lutropin response in this system did not parallel ovine lutropin. A counter-current distribution procedure for separation of rabbit lutropin subunits is described. Amino acid composition of the isolated subunits and intact rabbit lutropin was determined. The carbohydrate composition of the latter is presented; only amino sugar determinations are available for the subunits. The NH2-terminal amino acids are phenylalanine (alpha subunit) and alanine (beta subunit). Preliminary data on COOH-terminal amino acids are provided.


Solid-phase active site inhibitors of proteinases.

Phenylalanine chloromethyl ketone covalently attached to porous glass beads was synthesized to serve as a solid-phase active site directed inhibitor of chymotrypsin-like proteolytic enzymes. The solid-phase reagent inhibited 20 nmol of bovine chymotrypsin per gram of glass and covalently bound 30 nmol of protein per gram of glass. Sepharose-bound lysine chloromethyl ketones were synthesized to serve as inhibitors of trypsin-like enzymes. Sepharose-MethionylLysyl chloromethyl ketone inactivated and bound about 6.8 nmol of enzyme per ml of settled gel. In a preliminary experiment, a cyanogen bromide cleavage of the methionine residues showed that it should be possible to release all peptides but the peptide containing the active-site histidine. The immobilized trypsin was also reduced, carboxymethylated and digested with chymotrypsin. The potential of the solid-phase approach is in the isolation of a specific serine proteinase and in the sequence determination of residues surrounding the active-site histidine.


Synthesis, characterization and fluorescence studies on N-alpha-dansylalanyllysyl trypsino (HIS-46)-methane.

The 1-dimethylamino-5-naphthalene sulfonyl (dansyl) derivative of alanyllsyl chloromethane was synthesized and employed to introduce the fluorescent dansyl moiety specifically into the active site of trypsin via affinity labelling. The potential of dansylalanyllysyl chloromethane lies in its high degree of selectivity and markedly faster rate of enzyme inactivation when compared to previously synthesized, single residue affinity label chromophores. This permits the practical utilization of stoichiometric amounts of the inhibitor to achieve 100% inactivation of trypsin, even at high dilutions. The transfer of energy between the four tryptophan residues of trypsin and the bound dansyl group has been investigated in the fluorescent inhibitor-enzyme conjugate. From transfer efficiency measurements mean distances of 19.0 A and 19.3 A between the point of attachment of the dansyl group and the four tryptophan residues of trypsin have been calculated. These compare well with the mean value of 18.8 A derived from calculations based on crystallographic model studies.


The canine eye: the vessels of aqueous drainage.

Lumenal castings of canine vessels of aqueous drainage, prepared by microvascular injection of room-temperature vulcanizing silicone or Batson's methacrylate corrosion compound No. 17, were viewed with a stereo dissecting microscope and by scanning electron microscopy. The pathways for aqueous drainage in the dog appear to extend radially from the interstices of the filtering meshwork, through the deep intrascleral venous plexus, to the midscleral venous circle of Hovius, which also serves choroidal venous drainage. Venous blood mixed with aqueous humor may egress the globe from the venous plexus of Hovius via the anterior ciliary vein or posterior vortex veins. The canine pathways of aqueous drainage are distinguished from those in the primate by the absence of Schlemm's canal and the intrascleral mergence of drainage routes for aqueous humor and uveal venous blood.