Activity and Properties of Macrophage Migration Inhibitory Factor produced by Mixed Lymphocyte Cultures

THE macrophage migration test is an in vitro demonstration of delayed hypersensitivity. Supernatant fluids of sensitive lymphocytes cultured for 24 h in the presence of specific antigen contain migration inhibitory factor (MIF) that arrests the migration of macrophages of unsensitized animals in vitro¹,². In vivo, it induces delayed skin reactions³. The use of the macrophage migration test, based on differences of transplantation antigens in donor and recipient, to show histocompatibility has been suggested⁴. The test was also recommended as an indicator of immunological reactivity after organ transplantation, to demonstrate impending rejection⁵. It can demonstrate homograft sensitivity, for migration of peritoneal exudate cells (containing lymphocytes and macrophages) of CBA mice previously sensitized by grafts from A/Jax donors was inhibited when they were mixed with peritoneal exudate cells of the donor strain. However, histocompatibility was not demonstrated, for mixtures of peritoneal exudate cells of ungrafted CBA mice and A/Jax mice migrated regularly during the 24 h test⁶.     

Macrophage Migration Inhibition by Serum from Desensitized Animals Previously Sensitized with Tubercle Bacilli

MIGRATION of peritoneal exudate cells removed from guinea-pigs or mice exhibiting delayed hypersensitivity is inhibited by specific antigen^1–3. This in vitro macrophage migration inhibition has been regarded as a useful immunological test for delayed skin hypersensitivity^4,5. Studies of the mechanism of this phenomenon revealed that, in contact with specific antigen, lymphocytes from sensitized animals released into the medium a specific substance (migration inhibitory factor; MIF) capable of inhibiting the migration of normal macrophages^6,7. When injected intradermally into normal guinea-pigs, MIF elicits inflammatory reactions characterized by induration, erythema and mononuclear cell infiltration⁸.     

Detection of Antigen-binding Cells by Combined Rosette Formation and Autoradiography

DETERMINATION of the frequency of chromosome aberrations in cultured blood lymphocytes may provide a means of measuring ionizing radiation doses, at least after whole body exposure. Much work has been done with human blood irradiated in vitro^1,2, but before these results can be applied to radiation exposure in vivo, the difference between in vitro and in vivo exposure must be shown to be quantitatively negligible.     

Transfer of Immunity to Tumour Isografts by the Systemic Administration of Xenogeneic “Immune” RNA

SPECIFIC immunoreactivity can be conferred on lymphoid cells by incubation with RNA-rich extracts prepared from lymphoid tissues exposed to specific antigens in vivo¹ and in vitro^2,3. We have shown transfer of immunity to tumour specific antigens in vivo⁴ and in vitro⁵ by incubation of syngeneic spleen cells in vitro with RNA extracted from the lymphoid tissues of xenogeneic or syngeneic animals immunized with the tumour to be treated. Administration of these spleen cells to normal animals decreased the development and growth of isografts of the same tumour.     

Growth Response to Hormones by a New Rat Ovary Cell Line

SEVERAL endocrine cell lines established in recent years show a functional response to hormones in vitro¹ but, except for one mammary cell line², none of them exhibits the normal hormone requirement for growth in vivo. We have now isolated a rat ovarian cell line whose growth in vitro is markedly stimulated by bovine luteinizing hormone (LH-NIH-B7), a pituitary gonadotrophin and by dexamethasone, a synthetic glucocorticoid. This cell line provides the first permanent in vitro system for studying the growth stimulation of gonadal cells by hormones.     

<Emphasis Type="BoldItalic">In vivo</Emphasis> conditions influence the coding of stimulus features by bursts of action potentials

The functional role of burst firing (i.e. the firing of packets of action potentials followed by quiescence) in sensory processing is still under debate. Should bursts be considered as unitary events that signal the presence of a particular feature in the sensory environment or is information about stimulus attributes contained within their temporal structure? We compared the coding of stimulus attributes by bursts in vivo and in vitro of electrosensory pyramidal neurons in weakly electric fish by computing correlations between burst and stimulus attributes. Our results show that, while these correlations were strong in magnitude and significant in vitro, they were actually much weaker in magnitude if at all significant in vivo. We used a mathematical model of pyramidal neuron activity in vivo and showed that such a model could reproduce the correlations seen in vitro, thereby suggesting that differences in burst coding were not due to differences in bursting seen in vivo and in vitro. We next tested whether variability in the baseline (i.e. without stimulation) activity of ELL pyramidal neurons could account for these differences. To do so, we injected noise into our model whose intensity was calibrated to mimic baseline activity variability as quantified by the coefficient of variation. We found that this noise caused significant decreases in the magnitude of correlations between burst and stimulus attributes and could account for differences between in vitro and in vivo conditions. We then tested this prediction experimentally by directly injecting noise in vitro through the recording electrode. Our results show that this caused a lowering in magnitude of the correlations between burst and stimulus attributes in vitro and gave rise to values that were quantitatively similar to those seen under in vivo conditions. While it is expected that noise in the form of baseline activity variability will lower correlations between burst and stimulus attributes, our results show that such variability can account for differences seen in vivo. Thus, the high variability seen under in vivo conditions has profound consequences on the coding of information by bursts in ELL pyramidal neurons. In particular, our results support the viewpoint that bursts serve as a detector of particular stimulus features but do not carry detailed information about such features in their structure.     

Mutant tRNA<Superscript>His</Superscript> Ineffective in Repression and Lacking Two Pseudouridine Modifications

TRANSFER RNA has been implicated in the regulation of a number of amino-acid biosynthetic operons^1–4. Histidyl-tRNA^His has been shown to be involved in regulation of the histidine operon by analysis of six genes (hisO, hisR, hisS, hisT, hisU, hisW), mutation of which causes derepression of the enzymes of the histidine biosynthetic pathway in Salmonella typhimurium^5–7. A class of derepressed mutants (hisR) has only about 55% as much tRNA^His as the wild type⁴ and in the one example sequenced, contains tRNA^HIS with a structure identical to that of the wild type⁸. Studies of mutants of the gene for histidyl-tRNA synthetase (hisS) indicated that the derepressed phenotype was associated with defects in the charging of tRNA^HISin vitro². The amounts of charged and uncharged tRNA^His present in vivo during physiological derepression of the wild type and in the six classes of regulatory mutants, have been determined⁹. This work has shown that repression of the histidine operon is correlated directly with the concentration of charged histidyl-tRNA^Hisin vivo and not with the ratio of charged to uncharged or the absolute amount of uncharged tRNA^His. The derepression observed in mutants, of hisS (the gene for histidyl-tRNA synthetase), hisR (the presumed structural gene for the single species of tRNA^His) and hisU and hisW (genes presumably involved in tRNA modification) may be explained by the lower cellular concentration of charged tRNA^His which these mutants contain.     

Studies on regulation of chorionic gonadotropin secretion in primates

The regulation of secretion of chorionic gonadotropin in primates has been studied using bothin vivo andin vitro models.In vivo studies using the pregnant bonnet monkey revealed that at the doses tested, the administration of progesterone or estradiol 17Β in combination or alone did not result in any appreciable change in the duration or magnitude of serum chorionic gonadotropin levels. However, administration of lutropin-releasing hormone by intravenous route resulted in significant increase in chorionic gonadotropin levels within 30–60 min and the extent of stimulation seemed to depend on the state of pregnancy. Forin vitro studies, explants or cells prepared from first trimester human placenta has been used. The functional integrity of these cells has been established by demonstrating the binding of [¹²⁵I]-labelled human chorionic gonadotropin antibody to the cells as well as the synthesis of [³H]-labelled human chorionic gonadotropin.In vitro studies using the cells revealed that addition of lutropin-releasing hormone caused a significant increase in chorionic gonadotropin and estradiol 17Β secreted into the medium. Thus bothin vivo andin vitro results suggest that lutropin-releasing hormone could be one of the factors involved in regulation of chorionic gonadotropin secretion in primates.     

Effect of Rifampicin on the Development of Tumours induced by Adenovirus in Male Hamsters

INITIAL in vitro studies established that rifampicin, one of a group of rifamycin SV derivatives^1,2, prohibits bacterial growth and phage replication by binding to a polypeptide component of the microbial DNA-dependent RNA polymerase^3–7. The trachoma agent and related psittacosis-lymphogranuloma agents are also inhibited in vitro and in embryonated eggs by this drug⁸. Further studies have shown that rifampicin is active against a number of bacteria in vivo, both after parenteral and oral administration^1,9,10. It also inhibits malaria in mice¹¹ and trachoma in monkeys^12,13 and is of special value in the treatment of human tuberculosis^14–16. The low toxicity of the rifamycins in mammals¹⁷ has been attributed to the observed relative insensitivity of mammalian RNA polymerase to the rifamycins in vitro^3,18.     

Stereochemistry of Intercalation: Interaction of Daunomycin with DNA

DAUNOMYCIN^1–3, a glycosidic anthracycline antibiotic from Streptomyces peucetius⁴, is being used in the treatment of acute leukaemia and solid tumours in man^5,6. The biological activity seems to be due to complex formation with the DNA of deoxyribonucleoprotein⁴. In vivo, daunomycin inhibits both RNA and DNA synthesis^7,8 and, in vitro, DNA-dependent RNA polymerase and DNA polymerase^7–9.     

Cell Interactions in Humoral and Cell-mediated Immunity

THYMUS-derived lymphocytes (T-cells) play an important role in the initiation of both humoral and cell-mediated immunity^1–3. We have investigated whether the helper function and the cell-mediated killer function of lymphocyte populations are performed by the same cells, by assaying thymus-derived lymphocytes both for their capacity to cooperate in vitro with bone marrow-derived lymphocytes (B-cells) in the induction of plaque-forming cells and for their capacity to cause in vitro complement independent lysis of target cells.     

Effects of α- and β-Ecdysone on <Emphasis Type="Italic">in vitro</Emphasis> Diploid Cell Multiplication in <Emphasis Type="Italic">Drosophila melanogaster</Emphasis>

THE moulting hormone of insects is sometimes referred to as a growth and differentiation hormone. The steroid ecdysone and ecdysone analogues were shown to promote differentiation both in vivo and in vitro^1–3 but their action on cell multiplication is not certain^4–10. I used a diploid cell line, established from Drosophila melanogaster embryos by Echalier and Ohanessian^11,12, to assay insect hormones.     

Lidocaine Transdermal Patch: Pharmacokinetic Modeling and <Emphasis Type="Italic">In Vitro</Emphasis>–<Emphasis Type="Italic">In Vivo</Emphasis> Correlation (IVIVC)

The present study aims to develop the correlation between in vitro and in vivo skin permeation of lidocaine in its transdermal patch. In order to minimize the run-to-run variability during in vitro skin permeation studies, release normalized cumulative percent (%Ct _n) was calculated. A suitable polynomial mathematical model was used to establish a correlation between time and %Ct _n. Percent in vivo absorbed was calculated by using numerical deconvolution (NDC) and non-compartmental analysis (NCA) methods. Pharmacokinetic (PK) parameters such as AUC _last and C _max were predicted with the established in vitro–in vivo correlation (IVIVC) models. The minimum prediction errors in NDC method for C _max were found to be ?30.9 and ?25.4% for studies I (in vivo study in human volunteers with one batch of Lidoderm patch; internal validation) and II (in vivo study in human volunteers with another batch of Lidoderm patch; external validation), respectively, whereas minimum prediction errors in NCA method were relatively low (3.9 and 0.03% for studies I and II, respectively) compared to those in NDC method. The prediction errors for AUC _last were found to be less than 2% for both methods and studies. The established method in this study could be a potential approach for predicting the bioavailability and/or bioequivalence for transdermal drug delivery systems.     

Characterisation of<Emphasis Type="Italic">fusarium graminearum</Emphasis> and<Emphasis Type="Italic">F. culmorum</Emphasis> isolates by mycotoxin production and aggressiveness to wheat

A total of 27Fusarium culmorum isolates from Germany and 41F. graminearum isolates from Kenya were investigated for aggressiveness and mycotoxin production on wheat ears. In addition, ergosterol content of the kernels from ears inoculated withF. graminearum was determined and theF. culmorum isolates were tested for mycotoxin productionin vitro. For both pathogens, isolates markedly differed in aggressiveness. 59% and 37% of theF. culmorum isolates produced NIV and DON, respectively,in vivo andin vitro. The DON-producing isolates also produced 3-acDONin vitro. The more aggressive isolates produced mainly DON while the less aggressive isolates produced mainly NIV. 12% and 85% of theF. graminearum isolates produced NIV and DON, respectively. The highly aggressive isolates produced higher amounts of DON, aggressiveness being highly correlated to DON content in the kernels. NIV-producing isolates were less aggressive. Ergosterol content of kernels was moderately correlated to aggressiveness but highly correlated to DON content. Disease severity was associated with kernel weight reduction.     

Fusion <Emphasis Type="Italic">in vitro</Emphasis> of Membranes from Animal Cells

THE biological implications of membrane fusion are of general importance and several publications deal with the theoretical and practical aspects of this phenomenon^1–3. In this report, we describe experiments on the in vitro association or fusion of membranes. Because fusion was observed under physiological conditions it is possible that similar interactions also occur in vivo.     

<Emphasis Type="Italic">In Vivo</Emphasis> Evaluation of a PEO-Gellan Gum Semi-Interpenetrating Polymer Network for the Oral Delivery of Sulpiride

In this study, an optimized epichlorohydrin-crosslinked semi-interpenetrating polymer network xerogel matrix system (XePoMas) for the controlled delivery of sulpiride was prepared. The ability of XePoMas to sustain drug release was determined by in vitro and in vivo drug release experiments. Swelling of the xerogel over the 24-h experimental period ranged from 346 to 648%; swelling was observed to increase exponentially over the initial 8 h. In vitro drug release depicted a linear zero order drug release profile with an R ² value of 0.9956. The ability of the fabricated XePoMas to sustain drug release and enhance bioavailability of sulpiride in vivo was investigated by evaluating the plasma drug concentration over 24 h in the large pig model. The optimized XePoMas formulation was shown to increase intestinal absorption of sulpiride to a greater extent than the marketed product in vivo, with a C _max of 830.58 ng/mL after 15 h.     

On the Mechanism of Action of <Emphasis Type="Italic">lac</Emphasis> Repressor

Chen et al. have proved conclusively that lac repressor and RNA polymerase bind independently to wild type lac DNA in vitro. To explain the lacp ^s mutation, which causes competitive binding between repressor and polymerase, they suggest that a new promoter site has been created near the lac operator.     

Inhibition of Adenosine Uptake by Platelets

ADENOSINE inhibits platelet aggregation induced by adenosine diphosphate both in vivo and in vitro^1,2. In platelet-rich plasma, inhibition by adenosine first increases and then decreases^1–6, presumably because adenosine is either incorporated into the platelet and converted to nucleotides, or degraded in the plasma by adenosine deaminase (ADA) to inosine and then hypoxanthine^6,7.