Difficulties in the diagnosis of bacterial meningitis. Evaluation of antibiotic pretreatment and causes of admission to hospital. In 104 patients with bacterial meningitis admitted to departments of general medicine, the diagnosis before admission and the effect of previous antibiotic treatment were studied. Antibiotic therapy begun before admission in 30 patients did not affect the ability to make a bacteriological diagnosis. The duration of illness before admission was longer in the treated than in the untreated group. About 50% of the patients were admitted with diagnoses other than meningitis, although half these patients had distinct meningeal signs on arrival at hospital. In the whole series, 76% had meningeal signs on arrival. On the other hand, 95% of 108 patients with lymphocytic meningitis were admitted with a diagnosis of meningitis. The only significant clinical finding in the wrongly diagnosed group was a temperature higher than 40 degrees C on admission to hospital. The diagnosis before admission was not related to the treatment given. The findings illustrate the difficulties of diagnosing bacterial meningitis in the home.


[Acute pancreatitis. Aspects of the pathophysiology (author's transl)]. Three aspects of the pathophysiology of acute pancreatitis are discussed: 1. the initiating mechanisms, 2. the mechanisms of the fat necrosis, 3. the processes leading to shock phenomena. It is pointed out that the intraglandular activation of the precursors for both lipolytic and proteolytic enzymes seems to be essential for the initiating mechanisms of the disease. The role of the hormone dependent lipolytic enzyme of the fat tissue is discussed in relation to the occurrence of extrapancreatic fat necrosis. The role of the vaso-active compounds, such as plasma kinins and histamine for the occurrence of shock during acute hemorrhagic pancreatitis is pointed out.


Inhibition of oxidation by peroxidase of human serum proteins. The oxidation of essential serum proteins, albumin and gamma globulin, by the enzyme peroxidase can be partially inhibited by compounds, such as EDTA and 2,4-pentanedione, that complex with the iron ion in peroxidase. The importance of such inhibition lies in the circumstance that the oxidations in question might be a possible causative factor in tissue aging.


Increase of serum very low density lipoproteins in rats after administration of alpha-hexachlorocyclohexane. After enteral administration of 200 mg/kg alpha-hexachlorocyclohexane (alpha-HCH) female Wistar rats develop a hyperlipemia. 48 h after administration of alpha-HCH, serum triglycerides are increased by 300%, whereas both serum cholesterol and serum total phospholipids only increase by about 45%. Serum free fatty acids are not significantly altered. Fractionation of the serum lipoproteins by ultracentrifugation shows that the hyperlipemia is due to a fivefold increase in serum very low density lipoproteins. Hepatic triglyceride secretion, calculated after i.v. injection of Triton WR 1339, is increased in animals pretreated wtih alpha-HCH. Corresponding to this observation, drugs known to diminish the triglyceride secretion of the liver, such as actinomycin D, cycloheximide; glucagon, orotic acid, CFT 1201, and CFT 1042 reduce the alpha-HCH-induced hyperlipemia. We concluded from the results that hyperlipoproteinemia after alpha-HCH is due to an increased hepatic very low density lipoprotein secretion. At the same time, the blood sugar level was decreased in fasting animals after treatment with alpha-HCH. Earlier experiments suggest that this effect is due to a decreased gluconeogenesis in the liver.


Effect of hypophysectomy on the stimulation of liver growth by alpha-hexachlorocyclohexane, phenobarbital, and partial hepatectomy in the rat. alpha-Hexachlorocyclohexane (alpha-HCH) or phenobarbital (PB) elicit growth and cell multiplication in rat liver. In hypophysectomized rats, alpha-HCH and PB induce an increase in liver mass, but no increase in liver DNA. Hypophysectomy without additional treatment results in a decrease of liver size and RNA, while the DNA content remains unchanged, thereby leading to a relative DNA surplus. 1/3-hepatectomy in hypophysectomized animals leads to a small increase of hepatic DNA only; after 2/3-hepatectomy 75-80% of the original liver DNA are restored. In rats with intact hypophysis losses of liver DNA are known to be restored completely. The findings suggest that the relative DNA surplus in hypophysectomized rats prevents the stimulation of DNA synthesis by weak growth stimuli such as alpha-HCH, PB, and 1/3-hepatectomy. If the relative DNA surplus is eliminated by partial hepatectomy, the inducers do produce DNA multiplication. It is concluded that the induction of liver growth and cell multiplication by alpha-HCH and PB does not require the presence of the hypophysis or one of its hormones.


[Pulmonary fibrosis due to bleomycin].

[Pulmonary fibrosis due to bleomycin]. Being little myelotoxic, Bleomycin (BLM) is more and more used in association with other anti-neoplastic drugs; its pulmonary toxicity is not to be overlooked and here is a recall of the risks of pulmonary fibrosis during treatment by BLM. This review is based on a personal experience of 45 cases, of which 6 were cases of respiratory insufficiency. It derived also from a study of the literature which expressed the frequency of respiratory accidents in figures varying from 2 to 94% according to the criteria of tolerance used by the authors. BLM toxicity for the pulmonary tract being important some precautions should be taken in its use: specially supervising aged patients or those with a pastpathological respiratory story; it should particularly concern the functional side, using CO ductance to trace the first signs of toxicity A total dose of 450 mg should not be exceeded.


[Severe acute pancreatitis; course and response to intensive therapy (author's transl)]. Two cases of severe pancreatitis and deep shock are reviewed. In the first case measures to relieve the shock state were unsuccessful. Laparatomy was, therefore, performed and revealed acute haemorrhagic pancreatitis with diffuse purulent peritonitis. After insertion of a drain and irrigation of the abdominal cavity the incision was closed. Twice-daily lavage of the pancreatic fistula via a drain was continued until, after 5 months, the fistula healed spontaneously. In the second case intensive therapy succeeded after 9 days in controlling the acute stage of the disease. Conservative treatment was continued for 6 weeks and the patient was then discharged from hospital. He was re-admitted 3 weeks later because of suspicious clinical and biochemical signs of obstructive jaundice. Laparatomy disclosed inflammatory stenosis of the distal portion of the common bile duct and Vater's papilla and also a pancreatic pseudocyst the size of a child's head. The latter was removed and a drain was inserted. There were no postoperative complications.


alpha-Fetoprotein is not a component of the estradiol receptor of the rat uterus. In high-salt medium, cytosol from immature rat uteri displays two main high-affinity estradiol-binding peaks after ultracentrifugation in a sucrose gradient. The two components are the estradiol receptor which has a sedimentation coefficient of 5.5 S, and the alpha-fetoprotein which sediments at 4.5 S. The dissociation rate constants (k-1) of plasma alpha-fetoprotein-estradiol complexes measured at 0 degrees in the absence or presence of 0.4 M KCl were found to be 7 X 10(-5) and 8 X 10(-5) sec-1, respectively. The half-time of dissociation of these hormone-plasma protein complexes is 100-200 times more rapid than that of the  estradiol-receptor complexes. These data led to the use of two "differential dissociation" methods for the measurement of the hormone-binding protein complexes. In a high-salt cytosol, the charcoal technique measured selectively the receptor binding sites; the hydroxylapatite technique measured the sum of the alpha-fetoprotein plus receptor binding sites. Under these conditions, binding specificity studies provided evidence that alpha-fetoprotein is not a subunit of the receptor. This was confirmed by binding specificity studies in high-salt medium of the receptor separated from alpha-fetoprotein by ultracentrifugation.


Terminally repeated sequences in the avian sarcoma virus RNA genome. The initiation of DNA synthesis in vitro by RNA-directed DNA polymerase (deoxynucleosidetriphosphate: DNA deoxynucleotidyltransferase, EC 2.7.7.7) of avian oncornaviruses requires a tRNAtrp primer molecule located close to the 5' end of the viral RNA genome. DNA transcripts, 100 nucleotides in length, initiated on the tRNAtrp primer molecule contain nucleotide sequences complementary to a large (25 nucleotides) RNase T1 oligonucleotide, T-13, located at the 5' terminus of the avian sarcoma virus RNA genome. tRNAtrp-initiated DNA transcripts with a length of about 70 nucleotides contain substantially fewer nucleotide sequences complementary to this 5'-terminal oligonucleotide, suggesting that the tRNAtrp primer associated with the avian sarcoma virus RNA is located approximately 100 nucleotides from the 5' end of the RNA. In addition, we present evidence to demonstrate that DNA transcribed from avian sarcoma virus RNA sequences located at the 3' end, immediately adjacent to the poly(A), contains nucleotide sequences that are complementary to the 5'-terminal T1 oligonucleotide T-13. These data indicate that the 5' end of the viral genome contains nucleotide sequences that are repeated at the 3' end of the genome. We conclude that the avian oncornavirus RNA genome is terminally redundant.


Structure of the gramicidin A channel: discrimination between the piL,D and the beta helix by electrical measurements with lipid bilayer membranes. Measurements with different chemically modified gramicidins in lipid bilayer membranes were used to discriminate between the dimeric pi(L,D) helix proposed by Urry and the dimeric parallel or antiparallel helices proposed by Veatch and Blout. Evidence for the pi(L,D) helix was obtained on the basis of the different actions of a negatively charged O-pyromellitylgramicidin and a negatively charged N-pyromellityldesformylgramicidin on lipid bilayer membranes. O-Pyromellitylgramicidin forms ionic channels in lipid membranes when it is applied to both sides of the membrane. In contrast to unmodified gramicidin, O-pyromellitylgramicidin is inactive when it is applied only to one side of the membrane. N-Pyromellityldesformylgramicidin does not form ionic channels in lipid bilayer membranes whether it is applied to one or both sides of the membrane. These results support the view that the gramicidin channel is formed by two pi(L,D) helices. Dimer formation by head-to-head association of two pi(L,D) helices needs six intermolecular hydrogen bonds, which are located at the formyl end of the molecule and which occur deep within the lipid membrane. In the head-to-head associated pi(L,D) helix the absence of the formyl group leads to an inactivation of the peptide, whereas in a parallel or antiparallel double-stranded helix the absence of the formyl group should have only minor effects.


Isolation of a type C RNA virus from an established human histiocytic lymphoma cell line. A type C RNA virus has been detected in the culture fluids of the SU-DHL-1 human histiocytic lymphoma cell line previously established in this laboratory. In electron micrographs, the virus closely resembled other typical mammalian type C RNA tumor viruses in size and morphology. Viral RNA-dependent DNA polymerase activity has been demonstrated in particles (densities of 1.15 and 1.22 g/ml) in the microsomal cytoplasmic fraction and in pellets of culture fluids. The enzyme is partially inhibited by antibodies to the RNA-dependent DNA polymerases of simian sarcoma virus and RD-114 virus but not by antibody to the polymerase of murine leukemia virus, suggesting some degree of relatedness to type C viruses of subhuman primate origin. Typical syncytial microplaques were induced when SU-DHL-1 cells were cocultivated with rat XC cells. Although no focus formation was noted in similarly cocultivated mouse UC1-B cell cultures, the numbers of foci induced in rat embryo fibroblasts by murine sarcoma virus were significantly increased by coinfection with the virus from SU-DHL-1 cell culture fluids. No other evidence of infectivity, inducibility, or capacity for helper rescue of defective murine sarcoma virus genomes has been detected to date in cocultivation studies with a spectrum of fibroblastic and other nonlymphoid indicator cell lines of human and other species of origin.


A defense of the use of metaphor in analytic theory formation. This study of psychoanalytic theory formation is based on the epistemology of Cassirer and Langer. It postulates the need of all sciences to operate with symbols of various levels of abstractions, including, in a very prominent way, metaphors. There is not just one scientific method. The current wave of criticism against psychoanalytic theory from within and without, and especially of its use of metaphor in theory formation, is based on a philosophy of radical empiricism which cannot do justice to the science of psychoanalysis.


[Benign and malignant tumors of the prostate (author's transl)]. Etiology, symptoms, diagnosis and treatment of carcinoma of the prostate and benign prostatic adenoma are discussed. In prostatic adenoma the treatment of choice is either open surgery or transurethral resection. Conservative medical treatment has not been successful. The decision between radical prostatectomy, radiotherapy, or hormonal treatment in prostatic carcinoma depends on the staging results of the disease including-if necessary-diagnostic lymphadenectomy.


[Prostatic gland arteriography. Vascular supply, diagnosis and differential diagnosis of adenoma and carcinoma of the prostate (author's transl)]. Vascular supply and architecture of the prostate are studied by 100 arteriographic surveys of the pelvis. The prostatic arterial supply can be differentiated in urethral and capsular branches. In a further group of 38 patients with prostate gland disease selective bilateral arteriographies of the internal iliac arteries were performed. In 18 patients with the clinical diagnosis of prostatic adenoma, arteriography detected a carcinoma of the prostate gland. These diagnoses were proven by microscopy. In 10 patients with clinically suspected carcinoma, 5 were diagnosed by arteriography as malignant and also confirmed microscopically. The predominant angiographic criteria of prostatic carcinoma are irregular spotted stain of the involved parenchyma, hypervascularity of the urethral vascular branches, and pathological vessels.


Evidence for two series of B-cell antigens in man and their comparison with HLA-D. Population and family studies of five sera with antibodies against B-cell determinants show an excellent fit of some of them with the HLA-D determinants, suggesting that the HLA-D determinants themselves or closely linked structures can be recognized by serology. One serum, Ag, might be coded for by a locus different from HLA-D, which could be located between HLA-A and -B. A two-color fluorescence test to detect B-cell antigens was used and found to save time and reagents and to give very good results.


Detection and quantitation of human Ia-type antigens with iodinated protein A and specific purification of antibodies against Ia-type alloantigens. Antibodies directed against specific human Ia-type antigens can easily be detected and quantitated by an improved radioimmunoassay using iodinated protein A bound to a specific antibody-Ia-antigen complex on the surface of freshly drawn peripheral human leukocytes, cultured human cell lines, or lymphoid cells fixed with glutardialdehyde or formaldehyde. The same principle can also be used for the detection of Ia alloantigens on human lymphocytes when testing them with specific antisera known to contain antibodies against transplantation antigens. These anti-Ia-alloantigen antibodies had been purified by a two-step procedure involving ion-exchange chromatography on DEAE-cellulose at pH 6.3 and the specific absorption on formaldehyde-fixed Ia-alloantigen-carrying homozygous cell lines, followed by elution of these antibodies with isotonic citrate buffer at pH 3.0. In this way an about 90-fold purification could be achieved. After such a purification the highly enriched antibody fraction still reacted selectively with one specificity of the Ia antigen system.


New groups and segregant series among B-cell alloantigens of the Merrit system. A study of leukemia cells, peripheral B cells, and lymphoblastoid cell lines. On the basis of reactions with a chronic lymphatic leukemia cell panel, evidence for 6 new specificities of the Merrit B-cell alloantigenic system in man is presented, bringing the number of provisionally defined specificities to 19. These can be roughly divided into two segregant series. The system is well represented on both homozygous and heterozygous B-type lymphoblastoid cell lines. In the cell lines some specificities show a suggestive but inexact correlation with HLA-D locus factors. This correlation is represented also on peripheral blood B cells, on the non-T variety of acute lymphatic leukemia cells, and on acute myeloid leukemia cells. Although great similarities exist, each of the cell populations may manifest differences other than or in addition to mere differences in B-cell antigen frequencies.


The study of human Aa-like specificities using antibody eluates from human lymphoblastoid cell lines. Five sera from multiparous women were absorbed and eluted from the Daudi cell line devoid of HLA antigens. These eluates recognized a specificity, Ly-Lil, of the Ly-Li system. Further absorption of one eluate in Bjab or Raji cell line subdivided this specificity. Family studies showed that the five eluates from the Daudi cell line segregate into two distinct groups.


Relation between HLA-DW and the b-lymphocyte specificities. Homozygous DW typing cells were tested for six B-lymphocyte specificities. All four of the second locus specificities of B lymphocytes were strongly associated with the DW specificities. DW1 typing cells were B group 6, DW2 were B4, DW3 were B5, and LD107 were B3. The first B-cell locus antigens 1 and 2 tended to be uniform within the DW groups. From an analysis of the typing responses of a panel of cells to the homozygous typing cells, it has become apparent that the first B-locus specificity present on the homozygous typing cells also plays a role in determining whether a typing response is obtained or not. Thus, the DW3 typing cells were themselves B2 and B5, and cells having B2-B5 were most frequently nonreactive to DW3 in mixed lymphocyte culture. Homozygous typing cells therefore mainly detect the second B-cell locus antigens and, to a lesser degree, the first locus specificities. Stated another way, homozygous typing cells do not define a single specificity, but rather the presence of two B-lymphocyte specificities, even though their responses often reflect matching of only the second B-locus specificity.


Human Ia-like antigens associated with HLA-d. Antisera raised with HLA-A- and -B-compatible, HLA-D-disparate combinations were cytotoxic to B lymphocytes from the immunizing donor's HLA-D phenotype. Four antisera recognized structures closely associated with the HLA-D determinants Dw2, Dw3, Dw4, and LD 108. One antiserum had a broad reactivity pattern, including Dw3, Dw6, and some unknown specificity(ies). In population and family studies these B-lymphocyte antigens behaved as if they were governed by one genetic locus in the B-D region of the HLA complex. Furthermore, the antisera were cytotoxic to a minor concavalin-A-reactive T-cell subpopulation. The antisera had previously been shown to inhibit the stimulating cells in mixed lymphocyte culture and to be capable of inhibiting the Fc receptor in the EA rosette assay. We conclude that the antisera produced by this method recognize Ia-like antigens closely associated with the HLA-D determinants.


B- and T-cell-specific alloantigens in man. Screening for B-cell-specific antibodies in unabsorbed pregnancy sera preselected for weak reactivity in regular HLA-A, -B, and -C screening yielded a relatively high proportion (35/81) of B-cell-specific antibodies. Most B-cell-specific antibodies react broadly, showing inclusion phenomena suggesting 'cross-reactivity' analogous to that observed in HLA-A and -B serology. In control experiments with T-cell-enriched suspensions three antisera reacted with T cells and not with B cells. These antisera are highly associated with HLA-A2 in the unrelated population and segregate with HLA haplotypes in families and with HLA-A in a family with HLA-A, B recombination, Thus it appears that the human equivalents of Ia antigens may include--in analogy to the murine Ia antigens--B cell- as well as T-cell-specific alloantigens.


Cytotoxic effect of anti-H-2 and anti-Ia antisera on human B and T cells. Anti-H-2 and anti-Ia alloimmune mouse antisera were tested by the microcytotoxicity test on human peripheral blood B- and T-cell preparations. Anti-H-2 antisera react by a higher titer and/or cytotoxicity scoring grade on B cels than on T cells. Anti-Ia antisera react practically only with B cells. It was assumed that anti-H-2 antisera contain two components. One component reacts specifically with certain HLA-A- or -B-locus antigens or other closley linked gene products. The other component reacts predominantly or only with B-cell determinants; the specificity of the latter component has not yet been studied in sufficient detail.


Cellular distrubtion, purification, and molecular nature of human Ia antigens. Human Ia antigens were extensively purified (1390-fold increase in specific activity) in 32% yield from BRI 8 cells, a lymphoblastoid B-cell line. Purification was monitored by using allogeneic antisera arising by foetal-maternal stimulation. The product, a glycoprotein fraction, contained the Ia antigens, the HLA-A and -B antigens, and a glycoprotein of unknown function. The glycoprotein fraction was composed of four glycosylated polypeptides with molecular weights of 43,000, 39,000, 33,000, and 28,000, and beta2-microglobulin; no polypeptide was linked to another by disulphide bridges. The A and B antigens only were absorbed by antibody against beta2-microglobulin. The Ia antigens comprised one each of the 33,000 and 28,000 molecular weight glycosylated polypeptides noncovalently linked together. Thus, only these chains were absorbed by xenogeneic anti-Ia antisera and were cross-linked by dimethyl-3-3'-dithiobispropionimidate dihydrochloride. The dimeric molecule bound deoxycholate (0.26 g/g of protein) and, when solubilized in deoxycholate, has a molecular weight of 77,000. The Ia allo- and xeno-antigenic activities were labile to heating and proteolysis and are probably determined by the polypeptide structure. Xenogeneic specific anti-Ia antisera were raised in rabbits and mice by immunizing with the glycoprotein fraction. These antisera reacted with B lymphocytes and monocytes but not T lymphocytes and fibroblasts. Their Fab fragments blocked the cytotoxicity of the allogeneic antisera for B lymphocytes and were potent inhibitors of the mixed lymphocyte reaction.


Typing an unrelated panel with PLT cells: association with DW clusters. Primed LD typing (PLT) cells prepared in one Laboratory (Madison) were shipped in the frozen state and tested in Tübingen on a separate panel that had been typed with homozygous typing cells. Those PLT cells that had been grouped, on the basis of their reaction with test cells of the Madison panel, as defining an HLA PL antigen showed identical or nearly identical patterns of reactivity with the Tübingen panel. Clear association between certain PL antigens and DW clusters as defined with homozygous typing cells could be demonstrated. Of particular interest may be combinations of certain PLT reactions with D-locus-typed cells, where the primed cells do not react as expected from the target's HLA-D type.


Relationship between Fc receptors and Ia alloantigens: analysis with a sensitive radioimmunoassay. This paper describes the successful use of 125I-labeled CgG aggregate to detect Ia-type alloantibodies in pregnancy sera. The blockade of aggregate uptake of a variety of normal mononuclear and leukemic cell types by anti-Ia alloantibodies is analyzed. Fc receptors and Ia alloantigens are clearly two distinct moleculr entities. The association between Fc receptors and Ia alloantigens on a quantitative level seems to depend on a ligand-binding mechanism to control their interaction rather than the presence of a topographical molecular tandem arrangement.


Primed LD typing: reagent preparation and definition of the HLA-D-region antigens. Primed LD typing cells were prepared against single HLA haplotypes within families and used to type a random panel of 48 individuals. PLT cells could be grouped on the basis of highly correlated responses with the panel test cells; the 21 different PLT cells could be used to define 6 different PL antigens. One of these, PL 3, was split by the responses of two of the 21 PLT cells which measured an antigen called PL 3.1.


Response of primed LD typing cells to homozygous typing cells. At least two different methods using cellular responses have been described for defining the determinants of the HLA-D region: typing with HLA-D homozygous cells and primed LD typing. Primed LD typing cells were generated in one-haplotype-different combinations and grouped on the basis of two or more cells appearing to define the same HLA-D-region-determined PL antigen. Such cells were restimulated with homozygous typing cells for several of the presently known HLA DW clusters. A very strong correlation was noted: PLT cells defining the antigen PL1 were restimulated with homozygous typing cells for DW3, those PLT cells defining the antigen PL2 were restimulated by homozygous typing cells for DW2, and those defining PL5  were restimulated by homozygous typing cells for DW1.


Detection of HLA-D clusters using primed LD typing. Using a set of 17 primed LD typing (PLT) cells tested on a panel of 35 unrelated cells, we showed that certain groups of PLT cells tended to detect similar unrelated cells. The PLT cells were grouped into seven clusters and these tended to correlate with the seven HLA-D specificities represented on the panel, as determined by HTC testing. These data suggest that the antigens that cause restimulation in PLT are similar to those HLA-D antigens detected by the homozygous typing cell (HTC) test or, alternatively but more unlikely, that the two typing methods are detecting genes in close linkage disequilibrium with the HLA-D region.


Cellology of HLA. I. The apparent homogeneity of HLA-D. We have demonstrated that there is a similarity in the distribution of restimulation responses of primed lymphocyte typing (PLT) cells raised between related and unrelated individuals, thus leading to the suggestion that HLA-D is a homogeneous determinant. It is proposed that within HLA a hitherto unrecognized heterogeneous structure may exist that is closely associated with HLA-D and which may give rise to intermediate reactions in primary mixed lymphocyte reactions and PLT.


Lack of restimulation by non-HLA antigens in primed lymphocyte typing. We have studied the role of non-HLA antigens in restimulation of PLT cells by the following protocols. Responder cells were sensitized to cells from family members differing by one HLA haplotype. The PLT cells were restimulated by family members HLA- or HLA-D-identical to the responder. In none of these cases did we find significant restimulation, indicating that it is unlikely that there is a restimulatory non-HLA locus with even two alleles (p less than 0.05) of equal frequency. Other protocols we have used are consistent with this interpretation. This indicates that antigens determined by genes segregating independently of HLA do not play a major role in the PLT restimulation.


Detection by three cellular immunological techniques of the antigenic determinants of the Ly-Li system, expressed on human B lymphocytes. A clear correlation was observed between the presence of an Ia-like antigenic B-cell system Ly-Li, detected serologically, and three cellular immunological techniques: [1] mixed lymphocyte reaction (MLR) inhibition by an anti-Li antiserum; [2] level of restimulation of anti-Ly-Li in-vitro-primed lymphocytes; and [3] detection of HLA-D alleles by homozygous typing cells. These results suggested that the allelic products detected serologically may be identical to those detected by the first two techniques, namely MLR inhibition and in-vitro-primed lymphocyte typing, and, possibly, HLA-D typing using homozygous typing cells, although the correlation was repeatedly found to be less clear for the last technique.


Selection of haploid spermatozoa and its application to HLA-D typing. The expression of HLA-A, -B, and -D determinants on the surface of spermatozoa is haploid. Using cytotoxicity and appropriate HLA antisera and complement, we were able to select haploid spermatozoa. The surviving sperms behaved similarly to homozygous lymphoid cells when cultured in mixed sperm lymphocyte culture (SLC). This method is being applied to define new HLA-D specificities, to improve typing and matching for bone marrow transplantation and kidney transplantation and to study their association with various diseases.


Population data on two new HLA-D determinants, EI and RE. Homozygous typing cells (HTC) for two new HLA-D determinants, EI and RE, defined by family studies, are described. The HLA-D typing experiments among more than 300 unrelated individuals showed phenotype frequencies of 0.045 for EI and 0.098 for RE. Since the tested population was also typed for its HLA-A and -B alleles, linkage disequilibrium parameters could be calculated: HLA-D type EI was statistically significantly associated with HLA-B13 and Bw17, HLA-D type RE with HLA-Bw40. These data support the working hypothesis that both EI and RE are new alleles of the HLA-D series.


MLC and CML studies in the family of a pair of HLA haploidentical chimeric twins. A pair of dizygotic twins of opposite sex were incidentally found to be erythrocyte chimeras. The chimerism was found to include the lymphocyte population as well, since the twins differed for the maternal while sharing one paternal HLA haplotype. In addition to lymphocytes carrying antigens controlled by the inherited maternal haplotype, each of the twins had a small fraction--about 15%--of lymphocytes of the other twin's type. Both twins are healthy and have normal children. Investigations of the family by mixed lymphocyte culture (MLC) and cell-mediated lympholysis (CML) tests showed that the two haplotypes by which the twins differed carried different HLA-D determinants; even so the lymphocytes of each twin were completely nonresponsive in MLC and CML to those of the other twin. No 'blocking' factors could be demonstrated in the serum of the twins. These results support Burnet's theory of clonal selection in fetal life.


Apparent lack of gene dosage effect in the PLT assay. In this paper the results of primed lymphocyte typing (PLT) tests are reported which show the response to HLA-D-specific stimulator and to unrelated cells. The aim was to find out the sensitivity of the PLT method for evaluating not only qualitative HLA-D-region differences but also gene dosage effects. In addition, the primed cells were restimulated with cells from HLA-D-homozygous individuals mutually negative with the specific stimulator in the usual 5-day primary mixed lymphocyte culture incubation. No gene dosage effect in PLT experiments could be observed.


HLA variants of cultured human lymphoid cells: evidence for mutational origin and estimation of mutation rate. Variants of a diploid lymphoid cell line that show a loss of HLA-B27 antigen occur randomly in time and independently of exposure to the alloantiserum used for their isolation. From these and previous findings of variant stability, inducibility by mutagens, and the absence of linked variation, we colclude that most HLA variants arise by mutation. The mutation rate for HLA-B27 loss is 8 x 10(-7) per cell per generation.


Autoantibodies to zona pellucida: a possible cause for infertility in women. Human and pig ovaries were tested by agar gel diffusion and found to contain several cross-reacting (common) antigens. At least one common antigen was located in the zona pellucida as determined by indirect immunofluorescence. Serum samples from 22 infertile women were tested on pig eggs by immunofluorescence, and six of these samples produced strong and nine produced moderate reactions with the zona pellucida. The autoantibodies may be responsible for infertility in these women.


Role of simple mastectomy in treating patients with breast cancer. From 1945 to 1971, 473 patients had simple mastectomies as the primary treatment for their breast cancer (421 had either a simple mastectomy alone [26%], a simple mastectomy followed by postoperative radiation therapy [64%], or preoperative irradiation followed by simple mastectomy [10%]). More than one half of these patients were "inoperable" by standard surgical criteria. The local recurrence rate was 24%, which compares favorably with the reported 48% local recurrence rate for "inoperable" patients treated by radical mastectomy. These recurrences were on the chest wall in two thirds of the patients and in the regional lymph nodes of the other third. No statistical differences in survival or recurrence rate could be correlated with the site of the primary cancers in the breasts or among the three treatment groups.


Evaluation of DursbanR and Dowco 214 as mosquito larvicides in rice-fields. Emulsifiable concentrates of DursbanR (chlorpyrifos) and Dowco 214 (chlorpyrifos-methyl) were tested as mosquito larvicides using Hudson knapsack sprayers on small plots of rice-fields on Penang Island. The mosquitoes found in these rice-fields are predominantly Anopheles campestris and Culex tritaeniorhynchus. At dosages of 14, 28 and 56 gm hectare-1, Dursban is effective in maintaining the rice-fields free of Anopheles larvae for at least 2, 3 and 7 days respectively. Dowco 214 at 56 gm hectare-1 is able to keep the fields free of all mosquito larvae for at least 2 days.


Elevated CSF gamma globulins with cerebral "glioma.". A patient with a six-year history of seizures and a shorter  history of a progressive hemiparesis, had a glioma. A partial frontal lobectomy was performed. The cerebrospinal fluid, 38 days post-operatively, prior to irradiation, had an increase in gamma globulins. Following cranial irradiation the CSF gamma globulin level returned to normal and the hemiparesis disappeared. The significance of these CSF changes is discussed.


Effects of thrombin, chymotrypsin and aggregated gamma-globulins on the proteins of the human platelet membrane. Analysis of platelet membrane proteins and glycoproteins by SDS polyacrylamide gel electrophoresis was carried out before and after treatment with thrombin. Extended incubation with thrombin  (in the presence of EDTA or adenosine, which inhibit aggregation) produced extensive changes in the bands observed. With incubation times of a few minutes however, the changes were restricted to a glycopeptide, GP IV (approx. 90,000 Daltons) and one or two polypeptides of low molecular weight, in particular polypeptide 16 (approx. 23,000 Daltons). At 0--3 degrees C only polypeptide 16 was still hydrolyzed. Chymotrypsin, which does not activate platelets, attacked glycopeptides I, II, III but no changes were apparent in GP IV and polypeptide 16. When chymotrypsin-treated platelets were further incubated with thrombin, only GP IV and one to two low molecular weight polypeptides, especially polypeptide 16, were affected. As polypeptide 16 appears to be an integral membrane component it is possible that it, either by itself or in combination with GP IV, represents the primary thrombin substrate involved in platelet activation. Aggregated IgG, which also activates platelets, does not modify the membrane glycoproteins but does change the low molecular weight region in particular band 16.


The influence of chlormadinone acetate treatment on the concentration of some steroids in the blood, on the ovarian activity, and on the sexual behaviour of the mare. The effect of orally administered chlormadinone acetate (CAP), 10 mg daily for a period of 16 days, was investigated in the case of four mares with an irregular oestrous pattern accompanied by low ovarian activity (group 1), four mares which did not show oestrous symptoms at all and which had also low ovarian activity (group 2), and two ovariectomized and two ovario-hysterectomized mares (group 3). In all mares of group 1 and in the two ovariectomized mares of group 3 oestrus symptoms became apparent during treatment. Two mares of group 2 came into heat 8 and 11 days after the cessation of treatment. The two other mares of group 2 and the ovario-hysterectomized mares of group 3 showed no oestrous symptoms at all. In all except the two ovario-hysterectomized mares the progesterone concentration was found to increase in the peripheral blood from less than 1 ng/ml up to at least 4-6 ng/ml. In the mares which came into oestrus this progesterone increase occurred about three days before symptoms of heat became apparent. It is assumed that the uterus played an important role in the increase of the progesterone concentration in the blood during or after CAP treatment. A similar rise of the progesterone level was also found in the ovariectomized mares, indicating that this progesterone did not originate in the ovaries.


Reduction of error rates in the microlymphocytotoxicity test. The overall error rate of the microlymphocytotoxicity test has been reduced from 1.08% in 1971 to approximately 0.35%, based on comparisons of 882 replicate test pairs (different bleedings) and on family studies, with 0.11% attributable to "reading" errors. This reduction was attributed to automation for lymphocyte adding, better incubation temperature control, better technical preparation of cells and stronger, more monospecific sera. Three types of errors contribute to the overall error rate: random, sera-related (affected by strength and specificity), and lymphocyte-related (possible B-cell reactivity and donor/lymphocyte physiological condition). Strongly skewed error distributions, which seem to be lymphocyte-related, occurred in less than 4% of the test pairs.


HLA-D antigens in the Japanese population. A Japanese population was typed for HLA-D antigens for the first time using HLA-D homozygous typing cells Dw1 through Dw4 and the homozygous cells LD HO and LD AH, newly found in the Japanese population. Dw3 was absent from the Japanese population which also lacks B8. Dw1, Dw2 and Dw4 were found in this population, but these did not show significant linkage disequilibria with any alleles of the B or C loci. LD HO and LD AH were common in Japanese but absent in Caucasians. LD HO was found to be in strong linkage disequilibrium with Bw35. LD AH showed significant association with Bw22J.


Characteristic differences in inhibitory effects of oligodeoxyribonucleotides from DNA on human systemic lupus erythematodes (SLE) sera and rabbit anti-DNA sera. The difference in the antigenic determinant size of DNA for sera from patients with SLE and rabbit anti-DNA sera were investigated. Haptenic inhibition studies were carried out by measuring the inhibition of [3H]DNA-antibody binding by three different types of oligonucleotides which were prepared from formic acid-diphenylamine digests, hydrazinolyzed digests and pancreatic DNase digests, respectively. Oligonucleotides from DNase I digests showed potent inhibitory activity with both SLE sera and rabbit sera. However, the inhibitory activities of purine and pyrimidine oligonucleotides were more potent for SLE sera than for rabbit anti-DNA sera. The determinant size estimated for rabbit sera was in the range of tetra-to heptanucleotide, while in SLE sera it was in the range of di-and trinucleotide.


Chemical stabilization of Golgi silver chromate impregnations. Blocks of neural tissue were processed by a modified Golgi-Kopsch procedure and by the rapid Golgi method. Following the impregnation, the blocks were embedded in celloidin, sectioned at 100 micrometer, and collected in 70% alcohol. The sections were then processed as follows: 1) rinsed in distilled water; 2) substituted with 0.4M sodium bromide for five minutes; 3) reduced in Kodak D-19 developer; and 4) treated in 0.5M sodium thiosulfate. The silver chromate deposits within the impregnated cells are converted successively to silver bromide and to reduced silver by this procedure. Sections so treated resist decomposition of the Golgi impregnation, and they may be counterstained with conventional aqueous cresyl violet to demonstrate the cytoarchitecture of the Golgi-impregnated tissue.


Influence of mounting media on the fading of basic aniline dyes in epoxy embedded tissues. A simple cytophotometric technique is used to quantitate stain fading of basic aniline dye-stained epoxy-embedded tissues mounted in six different commonly used mountants. Significant fading was detected with all six mountants, although rates varied. The lowest rate of fading was observed with immersion oil and the highest rate of fading with Canada balsam. No significant differences in fading rates of four synthetic mounting preparations were observed.


Avidin binds to condensed chromatin.

Avidin binds to condensed chromatin. The protein avidin has been conjugated with fluorescein isothiocyanate and then used to treat cells and chromosome preparations. Fluorescence microscopy reveals that this protein is bound by areas of the specimen containing condensed chromatin.


Immunofluorescent localization of beta2-microglobulin in the human kidney. Using antisera to human beta2-microglobulin and an immunofluorescent technique, beta2-microglobulin was found to be localized along tubular and glomerular basement membranes of renal bipsies studied. Since beta2-microglobulin is a subunit of HLA preparations, it may also serve as an indirect marker for the presence of HLA antigens in these structures.


[Increase of the concentration of effector T-lymphocytes specific to antigens H2. 1. Elution of T-lymphocytes from the allogeneic target cells]. The technique of elution by pronase of murine lymph node cells adherent to relevant allogeneic target cells has been developed for enrichment of effectory T cell population. A fraction was obtained of killer cells possessing cytotoxic activity which exceeds that of the initial immune lymphocyte population by a factor of 6--8 as judged by the number of lymphocytes required for a 50% cytotoxic effect. The gain in CE is fairly reproducible in different H-2 systems and under various lymphocyte-target incubation conditions.


[Antigenic structure of the plasma membranes of mouse liver cells. III. Lipid haptens]. The total glycolipids, gangliosides, neutral lipids and some individual lipids of these fractions were isolated from the whole lipid extract of the mouse liver cell suspension. The antigenic activity of the isolated lipids was studied with the complement fixation test using the rabbit antisera against the liver cell ghosts. Active lipids were: glycosphingolipids, gangliosides, cardiolipin and fraction containing polar glyco- and phospholipid mixture. The cardiolipins isolated from animal cells of some species differed in their antigenic activity. With the immunofluorescence method, the absorbtion of antighost sera by lipid haptens resulted in a decreased fluorescence of plasma membranes of cells in the liver sections. This fact may evidence for the membrane localization of the studied haptens.


[Problem of the infertility of hybrids of the muscovy (Cairina moschata L.) and the domestic duck (Anas platyrhynchos domestica)]. Analysis of blood serum proteins electrophoretic spectra for perching and domestic ducks resulted in revealing interspecific differences of prealbumin and transferrin types showing incomplete identity of loci in analogous sites of some homologous chromosomes in the hybridizants. This confirms the genetic nature of sterility in this crossing combination hybrids whose blood serum proteins electrophoretic spectrum displays a complex hybrid phenotype.


Serum proteins in prostatic cancer VI. Reduction of the suppressive ("blocking"?) properties of serum on in vitro parameters of cell-mediated immunologic responsiveness following cryosurgery. Preliminary studies of sera from prostatic cancer patients have indicated a reduction in the presence of suppressive ('blocking'?) properties of in vitro parameters of cell-mediated immunologic responsiveness induced by a non-specific mitogen (phytohaemagglutinin) in association with a decrease in the level of alpha2-globulin and favourable clinical response following cryosurgery. The origin of the immunosuppression factor(s) migrating on electrophoresis in the alpha2-globulin fraction of serum remains to be identified. Earlier demonstration of suppression of leucocyte migration by factors elaborated from tumour cells and recent observations of the suppression of lymphocytic reactivity by seminal plasma and coaguloprostatic fluid suggest that suppression and reduction or abrogation of the suppressive properties of serum following cryosurgical destruction of tumour may be attributed to a reduction in soluble prostatic tumour-associated antigen shed into the circlation by previously viable tumour. Such antigen while not at a sufficient concentration to engender an immunologic response in the aging and tumour-burdened host, may, however, have been sufficient to pre-empt the effector limb of cell-mediated responsiveness contributing to the observed suppression of lymphocytic reactivity. Cryosurgery, resulting in necrosis and cell death with depletion of the primary source of antigen might thereby have permitted a previously overwhelmed host to respond, viz., the favourable clinical response observed.


Cytochrome staining of leukocytes in urine sediment. Assessment of inflammatory state. Urinary sediments with significant pyuria were supravitally stained to assess the inflammatory state. Clinic patients showed a significant correlation between acute urinary infection, inflammation, and the predominance of live polymorphonuclear leukocytes. Post-transurethral prostatic resection patients demonstrated to show a predominance of live polymorphonuclear leukocytes, which may be clinically usefull in a more careful selection before relying fully on bacteriologic cultures.


[Effect of blood serum proteins of healthy, newborn and carcinomatous rabbits on oxygen uptake by myocardium mitochondria]. Oxygen uptake by myocardium mitochondria in healthy rabbits and those suffering from cancer was studied as affected by blood sera and beta-globulin of newborn rabbits, pregnant female rabbits, healthy and carcinomatous rabbits. Respiration of myocardium mitochondria in healthy rabbits is inhibited under the effect of sera and beta-globulin of newborn rabbits and pregnant female rabbits. But the mentioned additives produce an opposite effect on oxygen uptake by myocardium mitochondria in the rabbit suffering from cancer as compared to myocardium mitochondria in the healthy rabbit. The difference in the protein action on oxygen uptake is levelled in the animals of different groups for mitochondria destructed under the effect of temperature and supersound.


[Intensity renewal of skin and serum soluble proteins in rats with vitamin A deficiency]. Intensity of skin and blood serum soluble proteins renewal under conditions of vitamin A deficiency in animal organism was studied with application of the label incorporation into the electrophoretic fractions of proteins 1,3 and 24 h after administration of 14C-lysine. Metabolism of collagen proteins, amino acid composition of skin soluble proteins, content of free amino acids and nucleic acid in skin were also examined. With vitamin A deficiency the intensity of 14C-lysine incorporation into the globulin fractions of serum proteins is 2-4 times as high and into the corresponding fractions of skin proteins is 3-5 times as low as in the control. Metabolism of gamma-globulins and collagen proteins of skin under conditions of vitamin A deficiency is found to be slow. The content of DNA and RNA in skin of the avitaminous animals is reliably 17 and 23% lower, respectively. In the skin extracts obtained by means of 0.15 M NaCl the content of free amino acids with vitamin A deficiency increases. No significant changes are found in the amino acid composition of skin proteins soluble in 0.15 M NaCl, with the exception for a 20% decrease in the content of cystine together with cystein.


A simple economical way to make Diazochrome slides ("blue slides") using a regular roentgen ray view box. A simplified method of making the so-called blue slide (Diazochrome slide) is described. Our method consists of five steps: (1) copying the original material with Kodalith film, (2) developing the Kodalith film, (3) exposing the Diazochrome film, in contact, emulsion to emulsion, with the Kodalith negative on the regular chest roentgen ray view box, (4) developing the Diazochrome film in the ammonia jar, and (5) mounting the finished film. A suggested exposure time for the Kodalith negative is one second at f/8 to f/11. A suggested exposure time for Diazochrome film is one hour, and development time for Diazochrome film is five minutes. One should, however, establish his own best exposure time of these films for his own setup. Our method is relatively simple, fast, economical, and convenient.


[Evolution of hepatocyte population in the process of chemical carcinogenesis]. By means of two different markers of differentiation, using immunofluorescene method, the authors have characterized changes in the population of hepatic cells 1-8 weeks following the start of 3'-methyl-4-dimethyl-aminoazobenzene or 2-acetyl-aminofluorene action. Alpha-fetoprotein served as a marker of embryonic hepatocyte differentiation; while ligandin-as a marker of high-differentiated mature hepatocytes. The toxic effect of the carcinogens on hepatic stem cells was accompanied with a decrease of ligandin content in centrilobular hepatocytes. Among newly proliferating elements "oval" cells, cells of bile tract epithelium and most of basophilic hepatocyte-like cells fail to contain either alpha-fetoprotein or ligandin. Small groups of basophilic hepatocyte-like cells would contein alpha-fetoprotein. In cells of high cylinder-shaped epithelium of intestinal type ligandin was found, but alpha-fetoprotein was not found. The latter was absent in oxyphilous hepatocytes of hyperplastic nodules. In terms of ligandin content three types of morphologically identical nodules were differentiated; a) ones not containing this protein, b) ones containing it in amounts common to normal mature hepatocyte, and c) hyperdifferentiated nodules containing abnormally high concentrations of ligandin. Within one nodule cell all cells were identical in ligandin content. Thus, it is shown that at early stages of chemical carcinogenesis there occure in the liver multiple foci of differentiation of various kind. These intensive processes are assumed to be essential for tumor evolution in the tissue.


[Binding of glucocorticoids by a transcortin-like protein in the cytoplasmic fraction of rat myocardium]. Myocardial cytosol of adult adrenalectomized rats bound specifically 3H-corticosterone and 3H-hydrocortisone and did not bind the synthetic 9alpha-fluoroglucocorticoid (dexamethasone) in vitro at 0-4 degrees. The specific binding of dexamethasone was not also found in the cytosol of neonatal rats. Heating of myocardial cytosol of adrenalectomized rats within 10 min at 60 degrees led to the complete loss of the glucocorticoid-binding activity. Treatment of the cytosol within the same period but at the temperature lower than 55 degrees did not affect its binding capacity. 3H-corticosterone-cytoreceptor complex was precipitated by ammonium sulphate at 40-60% of saturation. The complex was eluted from a column of Sephadex G-150 by solutions with both low (0.015 M KCl) and high (0.4 M KCl) ionic strength as the alone peak. This fraction corresponded by molecular weight to the transcortin complex. The data obtained and the results of our previous investigations suggest that myocardial glucocorticoid-binding cytoplasmic protein is related to transcortin-like cytoreceptors.


[Physicochemical characteristics of a specific beta 1-globulin in the blood serum of pregnant women]. Some physico-chemical properties of beta1-globulin (BGG) (the protein specific for pregnancy) were estimated. BGG was shown to be glycoprotein, possessing the antigenic individuality. The heterogeneity of the protein studied was observed in salt fractionation, polyacrylamide gel electrophoresis, ion exchange chromatography and isoelectrofocusing; this demonstrates the existence of various BGG fractions.


[Synthesis, physico-chemical and antigenic properties of the conjugated antigens of xanthine- and theophylline proteins]. Conjugated antigens of xanthine- and theophylline-protein were synthesized by the reaction of diazosalts of 8-aminoxanthine and 8-aminotheophylline with bovine serum albumin. UV-spectra of the synthesized preparations were studied and content of xanthine and theophylline, covalently bound with the substances, was estimated. Immunization of animals with the conjugates was found to cause the occurrence of antibodies in blood serum, which bound specifically various derivatives of purine.


[State of the kinin system and level of serum proteinase inhibitors in latent nephritis and the nephrotic syndrome of different etiology]. Main components of kinin system, the arginine-esterase activity and proteinase inhibitors were estimated in blood serum of patients with nephrotic syndrome of various etiology (glomerulonephritis, amyloidosis, systemic lupus erythematous) and also in patients with latent nephritis and in healthy donors. Content of all the kinin system components (kallikreinogen, kininogen and kininase 1) proved to be increased in all the forms of nephropathy studied. Free kallikrein was found in blood serum of patients with nephrotic syndrome as distinct from healthy persons and patients with latent nephritis. The arginine-esterase activity, which shows the level of trypsin-like proteinases, was altered dissimilarly, depending on the nephrotic syndrome etiology: it was maximally increased in nephrotic syndrome of amyloid genesis and decreased in patient with systemic lupus erythematosus. High content of kallikrein and kininase I with simultaneous decrease in kininogen was typical for patients with severe form of nephrotic syndrome. Impairment of kidney in nephrotic syndrome was also characterized by an increase in alpha1-antitrypsin and in the total antitryptic activity, which reached the maximal value in nephrotic syndrome of the I degree and decreased at the II degree of the disease. In nephrotic syndrome content of alpha2-macroglobulin was maximally increased at the II degree of nephrotic syndrome and decreased in severe form of the disease. The primary alteration in content of proteinase inhibitors and high level of kinin system components were assumed to determine the conditions for activation of kinin system in blood serum and to impair the nephrotic syndrome pathogenesis, which was complicated by systemic manifestations. High content of kinin system components was apparently determined by the increased synthesis in liver tissue in response to inflammation and massive proteinuria; kininase I and alpha2-macrolgobulin, as proteins with high molecular weight, were likely to be selectively retained in blood circulation when the capillary penetration was increased.


[Mechanism of effect and clinical use of antiandrogens]. It is reported on the possibilities of the application of anti-androgenics, especially of cyproterone acetate. The indication extends to hirsutism, sexual deviations, growth disturbances in pubertas praecox as well as diseases of the prostate. Particularly strong standard are to be applied in the treatment of fertile women, as there exists the danger of an intrauterine feminisation of male foetuses, when a pregnancy was not absolutely excluded. Side-effects and results of animal experiments are mentioned. The therapeutic mechanism of the anti-androgenics can be explained with the help of a concurrency mechanism at the androgen receptor or acceptor.


A fluorescence phase contrast study of the interaction between Toxoplasma gondii and lysosomes in living cells. Lysosomes of peritoneal macrophages, selectively pinpointed by aminoacridine-induced fluorescence, do not fuse with vacuoles containing viable Toxoplasma gondii RH strain.


[The influence of loss of blood and alcohol intoxication on early vital wound reactions (author's transl)]. The influence of loss of blood and alcohol intoxication on wound reactions in the first 12 hours after incisions into the skin of the back of guinea pigs was examined. As the most important result we found differences in the cellular and zymohistochemical reactions: while begin and progression of the dermal leucocytosis were not influenced, the parallel running activation of different structure bound enzymes was significantly delayed and decreased. Considering the first result and the haematogenous origin of the leucocytes there is reason to believe that in the hierarchy of the biological control circuit-at least in guinea pigs-the local reflex hyperaemia of the wound area certainly is primary to the dermal vasoconstriction during collapse. The animals with haemorrhagic shock showed a retardation in the liberation of histamine and serotonin. This decrease of the local mediators of the stromae dissociation is considered the reason of the impaired initial enzyme activities in the mesenchymal tissue.


[Distribution of blood in the kidney by death from exsanguination (author's transl)]. The distribution of blood in the human kidney in acute and protracted exsanguination has been examined by staining sections (30-50micro) with benzidine. After bleeding to death within a few minutes arteries, aterioles, glomeruli, and the capillary network in the renal cortex are empty. In protracted bleeding to death glomeruli are filled with blood, the vessels of renal cortex are empty except for some little arteries and some vasa afferentia. It may be possible to utilige the intrarenal blood distribution to reach an opinion as to the duration of the fatal hemorrhage and even to distinguish single or multiple episodes of hemorrhaging during the course of fatal exsanguination.


Investigation on the carcinogen burden by air pollution in man. XV. Polycyclic aromatic hydrocarbons in automobile exhaust gas--an inventory. Polycyclic aromatic hydrocarbons (=PAH) emitted by two different vehicles were separated by gas chromatography and characterized by mass spectrometry after enrichment from exhaust gas condensate. Gasoline was investigated also using this technique. The quantities of PAH with boiling points higher than 338 degrees C contained in the exhaust gas were compared to those present in the fuel. About 150 different PAH have been characterized by mass spectrometry, 75 of these were identified by means of comparison with authentic samples. Six compounds of the PAH-group consisting of 4-7 rings, which are suspected of possessing carcinogenic activity, are described here for the first time. Structures of these hitherto unidentified PAH are proposed by means of MS and UV spectral informations (11H-cyclopenta(qrs)benzo(e)-pyrene, 10H-cyclopenta(mno)benzo(a)pyrene, benzo(ghi)cyclopenta(pqr)perylene, cyclopentenopyrene). According to the different PAH-profiles of the fuel and of the exhaust gas, it is concluded that the main part of the PAH emitted is produced de novo during the combustion in the engine and furthermore that the PAH contained in the fuel are combusted for the most part.


Effect of fuel composition on the emission of phenols in the exhaust gas from a European car. The emission of phenols from a European car working with leaded and unleaded fuels with different percentage of aromatics has been considered. Fuels having the same aromatic content, but with a different composition of aromatic fraction, have also been taken into account. The results obtained showed that the emission of phenols increases with the increase of the aromatic content of fuel and also when unleaded instead of leaded fuels are used. The type of aromatic present in fuels was found to be important in forming the amount of both total and individual phenols emitted in the exhaust gas and in determining the number of phenolic compounds formed during combustion, although the phenol and isomer cresols were produced by combustion of all the fuels tested. The quantitative determination of individual phenols has been carried out on the benzene extract of the aqueous condensate and of the particulate matter of exhaust gas by the NaOH-extraction-GC-chromatographic method.


A new post-staining fixation technique for Acridine Orange. A technique for Acridine Orange staining of cells for automated flow analysis involving post-staining fixation with Millonig's glutaraldehyde buffer is presented. Preliminary data indicate that with this protocol there is decreased background fluorescence and increased signal to noise ration in flow. In addition, stained cells may be stored for several months without alteration in cellular morphology or fluorescence characteristics. Finally, nuclear and cytoplasmic fluorescence do not appear to vary significantly with respect to cell concentration or distribution on a slide or filter when cells are stained according to the glutaraldehyde protocol.


B and T cell markers of bone marrow and peripheral blood lymphoid cells in patients with paraproteinaemia. Studies have been carried out on B and T cells in bone marrow and peripheral blood from patients with paraproteinaemia. The peripheral blood of patients with multiple myeloma showed a significant increase of B cells, mainly lymphoid cells bearing immunoglobulins corresponding to the paraproteins, while in patients with benign monoclonal gammopathy only a slight increase of B cells and a moderate decrease of T cells have been found. As to the bone marrow, the B cell population was significantly raised in patients with multiple myeloma, but it remained unchanged in patients with benign monoclonal gammopathy. Our findings may offer a new possibility to distinguish between these two diseases and provide further data to their pathogenesis.


Treatment of chronic active hepatitis with either prednisolone or corticotrophin: a controlled trial. A total of 54 patients hospitalized for chronic active hepatitis were randomly treated, 29 with prednisolone (maintenance dose 15 mg/day) and 25 with depot synthetic corticotrophin (maintenance dose 1 mg/week, i.m.) and were followed up for 6 to 24 months or longer. In this series, young males predominated, the incidence of serum HBsAg positivity approached 80% in both treatment and no patient had initial evidence of cirrhosis or had autoimmune associated diseases. With either drug SGOT levels showed a decrease during the initial 12 months of therapy (p less than 0.05); initial jaundice, when present, had disappeared by the 3rd month of treatment. With both treatments globulins and gamma-globulins decreased significantly after 12 to 24 months of therapy. Serum HBsAg persisted in all but two cases. Serum liver biopsies showed the following evolutions of histological activity: 12 cases (22%) improved to the "inactive phase" (8 with prednisolone and 4 with corticotrophin); 19 (35%) improved to a lesser extent (8 with prednisolone and 11 with corticotrophin); 17 (32%) remained unchanged (11 with prednisolone and 6 with corticotrophin); 6 (11%) worsened (2 with prednisolone and 4 with corticotrophin). Morphological features of cirrhosis appearently developed in 15 patients (8 treated with prednisolone and 7 with corticotrophin) of whom 7 achieved improvement of histological aggressiveness concurrently. Differences between treatments were not significant. Side effects suggesting drug discontinuation occurred only in 6 cases.


[Alpha fetoproteins in serum of a patients with Hodgkin's disease]. Presence of alfa-fetoprotein was found in serum of patient in whom primary hepatoma was suspected. The post-mortem microscopical examination did not confirm the clinical diagnosis ut showed changes typical for Hodgkin's disease.


Observations on the morphology at the transition between the peripheral and the central nervous system in the cat. V. A light microscopical and histochemical study of S1 dorsal rootlets in developing kittens. The postnatal development of the transitional region (TR) i.e. the proximal free part of a spinal rootlet that contains both PNS and CNS tissue, was studied light-microscopically in semi-thin sections and after histochemical staining according to the Marchi and OTAN methods for the demonstration of degenerating myelin and according to the Gomori method for the demonstration of acid phosphatase activity. In the newborn kitten the PNS tissue extended well up to the spinal cord surface and the rootlets lacked a transitional region. The CNS tissue entered the root during the second postnatal week, and a trasitional region was fully established at the beginning of the second month. The degree of myelination in the group of large fibres differed on the two sides of the PNS-CNS borderline: well myelinated PNS fibres were transformed into poorly myelinated or apparently unmyelinated CNS-fibres. PNS and CNS myelin sheaths of large fibres appeared to be of equal thickness in the 4 week old kitten. During the first postnatal month large amounts of Marchi positive material and a high acid phosphatase activity occurred in complex paranodes and very short internodes in the PNS compartment just distally to the PNS-CNS borderline. In the adult cat Marchi positive bodies were numerous in the CNS compartment just proximally to the PNS-CNS borderline. The results are discussed against previous studies on focal demyelination as found during the normal development of the feline peripheral nervous system.


Histopathologic findings in surgical specimens of endolymphatic sac in Meniere's disease. Two types of abnormality in the endolymphatic sac acquired at shunt surgery are presented. One was deep brown pigmentation in the subepithelial connective tissue. The pigment did not react to Prussian blue, thus ruling out hemosiderin. The other, in seven out of eight specimens, was the perisaccular fibrosis. The fibrotic foci in four cases were assumed, from the findings under ordinary illumination and polarized light microscopy, to be reorganized scar tissue.


Klinefelter syndrome in the aged.

Klinefelter syndrome in the aged. Three cases of Klinefelter syndrome diagnosed late in life are reported with the clinical and post-mortem findings. The diagnosis was suspected owing to the absence of testes and the presence of associated varied clinical features. The urinary follicle-stimulating hormone levels were not elevated and were very low in two of the cases. Bronchopneumonia, ascending pyelonephritis and cystitis were the main causes of death but there were varied pathological findings in the prostate of benign hyperplasia, carcinoma and prepubertal gland. Marked atherosclerosis of the aorta and the lower-limb vessels were present but the coronary systems were little affected. It is suggested that, because of the presence of two X chromosomes and the absence of testes, this condition may give a female pattern of longevity, many examples being overlooked in old age on account of inadequate clinical examination. Some support for this hypothesis will be found if its incidence in the aged is shown to be greater than in the young.


His bundle electrocardiography in manifest and concealed right bundle branch extrasystoles. His bundle electrocardiography was helpful in the diagnosis of impulse formation in the right bundle branch. Ten patients with narrow QRS complexes had ectopic beats with an "incomplete" left bundle branch pattern and almost simultaneous activation of His bundle and ventricles. Both QRS morphology and H- - V intervals depended on the more proximal or distal location of the ectopic focus. In four patients with "complete" right bundle branch block the morphology of ectopic ventricular complexes and H- - V intervals also depeneded on the presence or absence of retrograde block and differential degrees of forward and/or retrograde conduction delays. Nine patients with "complete" right bundle branch block and four with "complete" left bundle branch block had premature beats which could have originated in the proximal right bundle branch, proximal left bundle branch, or distal His bundle. In one patient with "complete" left bundle branch block, "concealed" His bundle depolarizations (probably originating in an ectopic focus located in the right bundle branch) produced pseudo Type II (Mobitz) A-V block. Although lidocaine appeared to have been more effective in patients with bundle branch block than in those with narrow QRS complexes, further studies are necessary to corroborate this impression.


Percutaneous transthoracic needle aspiration of the lung. Diagnosing Pneumocystis carinii pneumonitis. Percutaneous transthoracic needle aspiration was performed on 228 occasions to obtain lung specimens from 202 patients with suspected Pneumocystis carinii pneumonitis. In 121 patients the diagnosis was established by identifying P carinii organisms in lung aspirates. Six patients whose aspirates did not contain P carinii were found to have the organism at autopsy. Findings from toluidine blue O and Gomori methenamine silver nitrate stains were equally satisfactory for detecting P carinii, but the percentage of specific diagnosis was higher when specimens were stained with both. Pneumothorax that required a thoracotomy tube occurred in 39 patients. Other infectious agents, either bacteria or fungi, were found in only four patients. Percutaneous pulmonary needle aspiration--when performed under fluoroscopic guidance--is a rapid and effective method for the diagnosis of P carinii pneumonitis.


Cytochemical "normal" and "abnormal" eosinophils in acute leukemias. Eosinophilic leukocytes may accompany a great variety of disorders and different types of acute leukemias. The most striking morphologic feature of eosinophils is their specific granules, but morphology alone often is insufficient to differentiate normal from abnormal eosinophils. Cytochemically, the eosinophils were considered "normal" when they did not contain alkaline phosphatase, chloroacetate esterase, toluidine blue metachromasia, Astra blue positivity, and specific PAS-positive granules, but did have peroxidase and cyanide-resistant peroxidase activities, Sudan black positivity and moderate naphthol-AS esterase or alpha-naphthyl esterase and acid phosphatase positivities. In seven cases of acute leukemias (two acute myeloblastic and five myelomonocytic), in contrast with their normal behaviour, the eosinophils show "abnormal" cytochemical positivities consisting of chloroesterase activity, PAS and Astra blue positivities of the specific granules, toluidine blue metachromasia, and cyanide-resistant peroxidase of a few specific granules. Cytochemical investigations may provide additional criteria for evaluating the abnormality of the eosinophilic cell in leukemias.


Prognostic factors for advanced diffuse histiocytic lymphoma following treatment with combination chemotherapy. Combination chemotherapy can dramatically improve the survival of those patients with advanced diffuse histiocytic lymphoma who enter complete remission. The age, sex, stage, constitutional symptoms, sites of disease and tumor mass of 56 consecutive patients with advanced diffuse histiocytic lymphoma were analyzed as prognostic factors. Twenty-four patients received cytoxan, vincristine, procarbazine and prednisone (C-MOPP) and 32 patients received bleomycin, adriamycin, cytoxan, vincristine and prednisone (BACOP). In 26 of 56 (46 per cent) a complete remission was achieved, in 21 of 56 (38 per cent) a partial remission and in nine of 56 (16 per cent) no response. Median survival with a complete remission was not reached at three years with 82 per cent still alive, but was significantly greater than with a partial remission (7.6 months median) or no response (3.2 months median). Survival of those with a partial remission and no response did not differ statistically. Stage IV disease, bone marrow involvement, gastrointestinal involvement and a tumor mass greater than 10 cm in diameter in a single location were all poor prognostic factors. No other analyzed factor significantly affected survival in this study. Complete response rates and survival curves for patients treated with C-MOPP were similar to those treated with BACOP.


Designing instructional media for severely retarded adolescents: a theoretical approach to trait-treatment interaction research. Moderately and severely retarded adolescents (N=70) individually viewed a cartoon, a live action film, and a pixilation film in color or black and white. A distractor method was used to measure frequency and duration of eye movement to visual stimuli. Content of visual stimuli was controlled, presentation of a distractor randomized, and all eye movement monitored to the nearest second. Black and white visual presentations had greater attentional value than color presentations and use of real human beings appeared to be more salient as an attention-holding technique than was use of animated characters. Implications of the results for use with severely retarded adolescents were discussed.


Isolation from human serum of an inactivator of bacterial lipopolysaccharide. By a series of chromatographic procedures involving precipitation by salt, gel filtration, anionic exchange, and hydroxyapatite elution, a protein--termed the lipopolysaccharide inactivator (LPS-I)--has been isolated from normal human serum. As a result of treatment of bacterial lipopolysaccharide (LPS) by LPS-I, the treated LPS loses its toxicity for mice and reactivity in the Limulus assay and appears to be irreversibly disaggregated. The inactivation of the LPS by the purified LPS-I is temperature and time dependent and is not blocked by the addition of irreversible inhibitors of serine esterases. The LPS inactivator migrates as an alpha-globulin in whole serum and has a sedimentation velocity of approximately 4.5S. Characteristics of the inactivated LPS are briefly described using internally labeled LPS.


The mast cell and eosinophil in nasal secretion. With an improvement of Hansel staining it became possible to identify mast cells, eosinophils and other cells in nasal smears. By this improved method 87% of children and 95% of adult patients who visited our allergy clinic proved to have mast cells in their nasal smears.


Acid phosphatases of the rat epididymis. II. Biochemical characteristics, subcellular distribution and histochemical localization. After separation of three epididymal acid phosphatases their biochemical properties were differently studied. With appropriate substrate and inhibitor selection the distribution of the enzymes in different segments as well as the subcellular fractions of the rat epididymis was also demonstrated. The same biochemical differences were also utilized in the histochemical localization of the enzymes. It was found that Enzyme I had a pH-optimum at 5.0, a molecular weight of 97 000 and Km-constant of 0.901 mM. It was highly sensitive to tartrate and fluoride and it was localized in lysosomes as well as in the epididymal spermatozoa. Enzyme II had an optimum at pH 5.7, a molecular weight of 67 000 and Km-constant of 0.806 mM. It was also inhibited by fluoride but more resistant to tartrate. Its subcellular site was also particulate, but it was also found in the epididymal fluid. Enzyme III had an optimum at pH 5.2, a molecular weight of 135 000 and Km-constant of 0.685 m. It was resistant to low concentrations of fluoride and tartrate but sensitive to heavy metal ions. The enzyme was soluble and it behaved incoherently in thermal inactivation. All enzymes revealed the highest activity in the thin middle segments of the epididymis. Histochemical naphthol substrates gave a diffuse reaction in the epididymal epithelial cells. With the lead salt methods glycerophosphates and p-nitrophenylphosphate gave somewhat different results depending on their specificity as substrates for the epididymal enzymes. Both substrates gave a strong reaction supranuclearly in the Golgi area of the chief cells. This activity was inhibited by tartrate and was most probably due to Enzyme I. The epididymal corpus and cauda showed additionally a very strong apical activity in the chief cells with p-nitrophenylphosphate. This activity was resitant to tartrate but sensitive to fluoride. It was concluded that this enzyme represents Enzyme II activity. Similar activity was also found in the dissolving "holocrine" cells of the corpus and the cauda. The activity of the soluble Enzyme III could not be revealed with the present methods and the spermatozoa in the tubular lumina remained unstained.


Analysis of serum proteins, using agarose electrophoresis in normal dogs and in dogs naturally infected with Dirofilaria immitis. The electrophoretic pattern of 130 serum samples from clinically normal dogs was evaluated, using agarose as the supporting matrix. The relative mobility of each globulin fraction in relationship to the mobility of albumin was determined in 70 dogs, and nomenclature based on the mobilities (Rf values) was proposed. Biuret protein determinations were done, and relative and absolute values of each serum protein fraction were determined. Changes in the fractions of serum proteins were evaluated in regard to sex and age of the dogs and the presence of microfilariae of Dirofilaria immitis, as determined by modified Knott tests.


Analysis of serum proteins and cerebrospinal fluid in clinically normal horses, using agarose electrophoresis. Using agarose as a supporting matrix, electrophoresis was conducted on 50 serum samples and 20 cerebrospinal fluid samples from clinically normal horses (n = 50) of various ages and breeds. The technique was shown to be reliable. A positive correlation between age and gamma-globulin concentration was found in young horses. Features of the electrophoretograms of serum and cerebrospinal fluid samples are discussed, and a nomenclature based on Rf values is proposed.


[Processes of spore formation and gramicidin C formation by Bacillus brevis var. G.B]. Correlation between gramidicin C biosynthesis and sporulation in the process of Bac. brevis var. G.B. cultivation under various aeration conditions was studied. It was shown that biosynthesis of gramicidin C was characteristic of the young cells and its level was the highest during the culture active growth. The time of the sporulating forms appearance depended on the aeration rate which defined the quantitative composition of the population during  the phase of the culture active growth and the stationary phase. Under the optimal aeration conditions the spore formation started during the phase of the culture active growth after some decrease in the maximum level of the cell productivity with respect to the antibiotic. When the aeration rate was increased the spore formation was shifted to later periods of the culture development, i.e. the stationary phase and the phase of the cell autolysis, the gap between the highest levels of gramicidin C buosynthesis and the beginning of sporulation being increased. Under certain aeration conditions the spore formation was not observed, while gramicidin C was synthesized. A conclusion has been made that there is no correlation between gramacidine C biosynthesis and sporualtion in Bacillus brevis var. G.B.


[Source of valine for protein V biosynthesis in a producer of actinomycin C]. The specific activity of 14C-valine in valyl-tRNA formed during incubation of the actinomycin C-producing organism with 14C-valine was constant and lower than that of the whole cell pool. The constancy of the valyl-tRNA was indicative of the presence of a separate compartment for the valine pool used for protein biosynthesis. A lower specific activity of valine in valyl-tRNA as compared to that of the whole cell pool may be indicative of a low rate of valine metabolism in such separate compartment with exogenic 14-valine or a higher concentration of free valine in it as compared to the specific activity of this amino acid at average per cell.


The direct radioimmunoassay technique for the determination of human alpha1-fetoprotein. The direct radioimmunoassay procedure for the detection of human alpha-fetoprotein is described. The technique requires 20 microliter of sample for a single estimation and includes ammonium sulfate precipitation. The whole assay can be completed in 18 hours. AFP concentrations ranging from 2 ng/m1 to 500 ng/m1 can be quantified with reproducibility sufficient for clinical as well as experimental purposes.


Alpha-globulin-like antigens in bacteria. Seventy-two strains of bacteria of the families Bacillaceae, Enterobacteriaceae and Micrococcaceae were investigated for antigens resembling Gm (1), Gm (2) and Inv (1) human immunoglobulin group factors. Antigens similar to these factors were encountered in some strains of the genus Bacillus. Staph. epidermidis strains had antigenic structures resembling Gm (1) and Inv (1) factors, and strains of Enterobacteriaceae structures resembling Gm (2) and Inv (1). Occurrence among various species of bacteria of antigens resembling human gamma-globulin group factors suggests immunization by bacterial infection as one of the causes of presence of anti-Gm and anti-Inv antibodies in human beings.


Preparation of anti-cea and anti-goat gamma-globulin sera for radioimmunologic assay of carcinoembryonic antigen. Goats were immunized with purified carcinoembryonic antigen, and the suitability of the antisera for clinical assays of carcinoembryonic antigen was characterized. Reactivity of equine sera to goat gamma-globulin as a precipitating factor in the radioimmunologic double antibody technique was also evaluated.


Cell culture studies with a cytopathic bovine rotavirus. The growth characteristics in MDBK cells of a calf rotavirus isolated in Northern Ireland are described. Of a range of cell cultures tested, the virus grew in secondary bovine kidney and MDBK cells, but consistently produced a CPE only in MDBK cells. The CPE consisted of cytoplasmic vacuolation, development of eosinophilic intracytoplasmic inclusions, and degeneration and detachment of cells from the monolayer. The onset of CPE was more rapid and its effects more severe in rolled cultures than stationary cultures. Immunofluorescent staining showed that the rotavirus was antigenically unrelated to avian and mammalian reoviruses. There was no significant difference in virus growth over the pH range 6.5 to 8.4.


Isolation of a phage lytic for several Brucella species following propagation of Tbilisi phage in the presence of mitomycin C. A new brucella-phage, designated D, was isolated from a batch of Tbilisi phage grown on Brucella abortus strain 544 in the presence of mitomycin C. Although resembling Tbilisi phage in morphology, it differed in its host range, adsorption properties, chemical stability and antigenic specificity.


Axoplasmic transport in ocular hypotony and papilledema in the monkey. Orthograde and retrograde axoplasmic transport were studied in optic nerve heads of seven hypotensive Macaca fascicularis eyes. Orthograde transport was studied by radioautography after intravitreal radioisotope injections. Retrograde transport was studied in the same eyes by horseradish peroxidase injection into the dorsal lateral geniculate nuclei or optic tracts. Three eyes had developed marked papilledema before injections. Orthograde axoplasmic transport was blocked in swollen axons of the optic disc anterior to Bruch membrane and in the lamina scleralis. Retrograde transport was blocked in axons within the lamina scleralis along the posterior edges of transverse scleral beams and in axons in the choroidal portion of the nerve head posterior to Bruch membrane. These results support the general concept that axoplasmic transport in the optic nerve head is sensitive to alterations in intraocular pressure, either increases or decreases. The edges of Bruch membrane and the openings in the lamina scleralis may constrict axon bundles in ocular hypotony.


Optic disc edema in raised intracranial pressure. IV. Axoplasmic transport in experimental papilledema. Tritiated leucine was injected intravitreously into the eyes of rhesus monkeys that had developed papilledema secondary to implantation of intracranial balloons. Autoradiographic studies of the optic nerve head showed that six hours after intravitreous injection of the isotope the fast component of axoplasmic transport accumulated in the regions of the lamina choroidalis and lamina scleralis. The slow component arrived at the optic nerve head two to four days after injection, and the swollen axons of the entire optic nerve head were filled with radioactive isotopes. Twelve days after injection of isotope, the axons in the optic nerve head were still diffusely labeled. Disturbance of axoplasmic transport was one of the primary events resulting in swelling of axons in papilledema. The pattern of axoplasmic disturbances in papilledema secondary to raised intracranial pressure was similar to that observed in papilledema secondary to ocular hypotony or increased intraocular pressure. Ocular hypotony, raised intracranial pressure, and increased intraocular pressure appear to share a final common pathway. All these conditions apparently converge into this final common pathway of disturbance of axoplasmic transport to give rise to papilledema.


Combined effects of bleomycin and X-rays on DNA synthesis in asynchronous Ehrlich ascites cells in suspension. Separate and combined effects of bleomycin and X-rays on rates of uptake of [14C]thymidine into Ehrlich ascites cells were assessed for extracellular drug concentrations of 12 micron (20 microgram/ml) and radiation doses of 2-5 krad. Rates of DNA synthesis were followed by monitoring the activity of the acid-insoluble portion of the asynchronous culture. The 14C activity of the acid-soluble pool was assessed in determining the rate of passage of 14C-TdR across the cell membrane. The results reveal that whilst the effects of each agent on TdR uptake rates are markedly different, they both inhibit DNA synthesis. Combined studies with both agents, 2-5 krad X-rays plus 20 microgram/ml bleomycin before, simultaneously or after exposure to X-rays, produced additive or less than additive effects on rates of incorporation of TdR into DNA. However, when the drug dose is split 2 X 10 microgram/ml before and after exposure to 2-5 krad X-rays, a synergistic effect on inhibition of DNA synthesis is observed.


The Duffy blood group determinants: their role in the susceptibility of human and animal erythrocytes to Plasmodium knowlesi malaria. Duffy blood group negative erythrocytes from blacks are refractory to invasion by Plasmodium knowlesi merozoites in vitro, and blacks with this genotype are resistant to infection by P. vivax in vivo. In order to evaluate in a direct manner the role of Duffy blood group determinants in invasion by P. knowlesi merozoites, we studied erythrocytes from three rare non-black Duffy negative individuals, Fy(a-b-), in whom the Duffy negative phenotype probably represents a mutation and not the introduction of the black Fy gene. These cells were resistant to invasion by P. knowlesi in vitro indicating that resistance to invasion is mediated by the FyFy genotype and not another closely linked factor. The erythrocyte receptors for invasion, however, may not be the Fya or Fyb Duffy antigens themselves, or at least not restricted to these determinants, since refractory Duffy negative human erythrocytes were invaded after treatment with trypsin or neuraminidase although these enzyme-treated cells still lacked Fy a and Fy b determinants. Furthermore, new world monkey erythrocytes and chymotrypsinized chimpanzee and kra monkey erythrocytes were invaded, although there was no serologic evidence of Fya or Fyb determinants on these cells.


Maternal serum alpha-fetoprotein and spontaneous abortion. The relationship between spontaneous abortion and maternal serum alpha-fetoprotein (AFP) levels was investigated between 9 and 25 weeks of pregnancy. Seven out of 126 (5-6 per cent) women who had spontaneous abortions had raised maternal serum AFP levels at their antenatal booking visit compared to 4 out of 247 (1-6 per cent) control patients who were delivered of single liveborn infants, a statistically significant differences. The raised AFP concentrations were, however, associated with spontaneous abortion only if the serum samples had been taken immediately before, or at sometime after the abortion was first clinically suspected. This suggests that high levels do not predict the development of abortion in women who have not already threatened to abort. It is therefore unlikely that women who have not already threatened to abort. Therefore, when maternal serum AFP levels are used to screen for fetal neural tube defects, women referred for a diagnostic amniocentesis on account of a high level are unlikely to have been selected on the basis of a tendency to abort.


Screening for fetal neural tube defects by maternal plasma alpha-fetoprotein determination. A total of 5539 consecutive pregnant patients at three maternity units in the City and Hackney District of London were screened for fetal neural tube defect by measurement of maternal plasma alpha-fetoprotein (AFP) levels. Only 25-7% of women booked at 16 to 22 weeks, the optimum time for this screening test; 54-1% booked before 16 weeks and 20-2% after 22 weeks. Of the women tested before 23 weeks, 300 had elevated levels of AFP in plasma and 14 of them had fetuses with abnormalities known to cause a rise in AFP levels (12 fetuses had a neural tube defect and 2 had alimentary tract abnormalities). Of women examined before 23 weeks, half of those with twin pregnancies had elevated levels of plasma AFP as did 16-7% of those who ultimately had a spontaneous abortion.


Evaluation of measurement of maternal plasma alpha-fetoprotein levels as a screening test for fetal neural tube defects. Maternal plasma alpha-fetoprotein (AFP) levels were estimated on 1844 consecutive patients at the time of their first hospital antenatal visits. There were 28 pregnancies with an 'abnormal' outcome: 19 patients had twins, 3 had an anencephalic fetus, and 6 a fetus with a spina bifida (4 of these had an open neural tube defect). Evaluation of maternal plasma AFP levels as a screening test for neural tube defects at 15 to 19 weeks gestation indicated a sensitivity of 50% taking the 'cut-off' point as the 95th centile of the plasma AFP levels in pregnancy with a normal outcome.


Omphalocoele and raised alpha-fetoprotein in maternal serum. Case report. We describe a pregnant patient with raised serum AFP levels. The infant had multiple congenital abnormalities which included an omphalocoele.


Serum proteinase inhibitors and other serum proteins in protein-energy malnutrition. 1. The concentrations of serum protein albumin, prealbumin and transferrin were determined in twenty-eight cases of protein-energy malnutrition (PEM) with infection, together with the levels of serum proteinase inhibitors (PI), alpha1-antitrypsin (AT), alpha1-antichymotrypsin (Ach), alpha2-macroglobulin (alpha2M) and inter-alpha-trypsin inhibitor (IalphaI). 2. Albumin, prealbumin and transferrin concentrations, as well as the levels of PI, IalphaI and alpha2M were found to be lower in cases of PEM associated with infection than the corresponding values for a group of healthy Thai preschool children and a group of newborn Thai children, but despite starvation AT and Ach values generally were increased. 3. The results provide support for the hypothesis that PI, especially AT and Ach might limit the synthesis of albumin, prealbumin and transferrin in PEM associated with infection, via the inhibition of the mobilization of body's own protein.


Proton nuclear magnetic resonances study of bleomycin in aqueous solution. Assignment of resonances. The 1H NMR spectrum of the glycopeptide antineoplastic antibiotic bleomycin has been examined in D2O solution (Fourier transform nuclear magnetic resonance, 270 MHZ) and in H2O solution (correlation nuclear magnetic resonance, 250 MHZ). Resonances have been assigned to specific hydrogens of the two most abundant congeners, bleomycin-A2 (BLM-A2) and bleomycin-B2 (BLM-B2), on the basis of (1) homonuclear spin decoupling, (2) comparison of the spectra of BLM-A2, BLM-B2, fragments of these antibiotics, and the related antibiotic phleomycin, and (3) the pH dependence of chemical shifts. Resonance assignments are presented for all the CH protons of BLM-A2 and BLM-B2 except for the saccharide groups, for which only the anomeric proton assignments are given. All of the NH protons have been identified with specific resonances except for the two primary amide groups, which yield four well-resolved peaks, whose specific assignment was not attempted. This study serves as a basis for future investigations of the conformation of bleomycin and its interaction with metals and nucleic acids.


Bleomycin-DNA interactions: fluorescence and proton magnetic resonance studies. The interaction of bleomycin A2 with DNA has been examined by fluorescence spectroscopy and proton magnetic resonance techniques. Fluorescence bands observed at 353 and 405 nm in the spectrum of bleomycin were assigned to the bithiazole and 4-aminopyrimidine rings, respectively. Quenching of bithiazole fluorescence by DNA was used to determine apparent equilibrium constants for the complex which, in 2.5 mM tris(hydroxymethyl)aminomethane buffer, pH 8.4, are 1.2 X 10(5) M-1 for bleomycin and 1.4 X 10(5) M-1 for tripeptide S, a partial acid hydrolysis product of the antibiotic. Uner these conditions, one molecule of bleomycin binds for every five to six base pairs in DNA. In the proton magnetic resonance spectrum of bleomycin, resonances emanating from the bithiazole rings and dimethylsulfonium groups are preferentially broadened and reduced in intensity in the presence of DNA, suggesting that these moieties bind most tightly to the polymer.


Quercetin: a novel inhibitor of Ca2+ influx and exocytosis in rat peritoneal mast cells. The effect of the transport ATPase inhibitor, quercetin on histamine secretion from antigen sensitized mast cells was examined. At micromolar concentrations, quercetin had an immediate inhibitory effect on histamine secretion mediated by antigen, concanavalin A and ATP but it had little effect on release induced by the ionophores A23187 and X537A. Quercetin exerts its effect after the binding of the releasing ligands and the distinction between its effect on ligand induced and A23187 induced secretion suggests that it affects the normal path of Ca2+ entry into the cell. The inhibitory effects of quercetin were compared with those of the structurally related anti-allergic drugs cromoglycate and AH7725.


Immunochemical studies on the subunits of rabbit-intestinal sucrase-isomaltase complex. Purified sucrase-isomaltase complex sucrose alpha-glucohydrolase, EC 3.2.1.48 - dextrin 6-alpha-glycanohydrolase, EC 3.2.1.10) solubilized by papain from rabbit intestine was dissociated by citraconylation into its subunits, sucrase and isomaltase, which were then isolated in a form active immunologically as well as enzymatically by affinity chromatography on Sephadex G-200 and gel-filtration on Bio-gel P-300. Antibodies against the purified complex inhibited isomaltase but not sucrase and formed precipitation lines, crossing each other, with isolated sucrase and isomaltase, showing that the two enzymes differ in antigenicity from each other. By absorbing the antibodies with isolated sucrase and isomaltase, antibodies specific for isomaltase and sucrase, respectively, were obtained. Like the original antibodies, both of the specific antibodies quantitatively agglutinated microvillous vesicles. Sucrase was inhibited by neither of the antibodies. In contrast, isomaltase was greatly inhibited by the isomaltase-specific antibodies, but not by the sucrase-specific ones.


New formamidase having substrate specificity for o-formylaminoacetophenone in pig liver. The supernatant (13 000 x g, 20 min) of pig liver homogenate was filtered with Sephadex G-200 and formamidase (aryl-formylamine amidohydrolase, EC 3.5.1.9)activity in each fraction was measured. When formylkynurenine was used as substrate, two peaks of formamidase activity were observed but, with formylaminoacetophenone as substrate, only one peak was observed. Formamidase in the lower molecular weight fraction is known as kynurenine formamidase (FA I), formamidase found here in the higher molecular weight fraction has not been previously reported. This form, designated FA II has been purified about 160-fold from pig liver. The formamidase obtained has substrate specificity for o-formylaminoacetophenone only and could not hydrolyze formylkynurenine. The optimal pH was 8.5 and the Km for o-formylaminoacetophenone was 1.66-10(-3) M. This formamidase was considered to be a new enzyme and was different from FA I in molecular weight and substrate specificity. This new formamidase was present in pig, rabbit and guinea pig liver and not present in rat or mouse liver.