Earlier Onset of δ-Retrovirus-Induced Leukemia after Splenectomy

Infection by δ-retroviruses such as human T-lymphotropic virus type 1 (HTLV-1) and bovine leukemia virus (BLV) is mostly asymptomatic. Indeed, only a minority (<5%) of δ-retrovirus infected hosts will develop either lymphoproliferative or neurodegenerative diseases after long latency periods. In fact, the host immune response is believed to tightly control viral replication but this assumption has not been definitely proven in vivo. Here, we provide direct experimental evidence demonstrating that integrity of the spleen is required to control pathogenesis. In the BLV model, we show that asplenia decreases efficiency of the immune response and induces an imbalance in cell dynamics resulting in accelerated onset of leukemia. These observations enlighten a potential threat in splenectomized HTLV-1 carriers and justify a regular preventive evaluation.     

On the course of thermal denaturation of deoxyribonucleic acids after γ-irradiation

Summary The changes which are caused by action of-irradiation onDNAs of various origin were followed by spectrophotometric method at differing thermal denaturation curves. It was found that all measured bacterialDNAs as well asDNA isolated from calf thymus, irradiated by exposures higher that 5×10⁴ R, produced significantly decreasedT _m values with concomittantly decreased hyperchromic effect and changed transition intervals obtained in 10^–2 M sodium phosphate, 10^–3 M EDTA medium at pH 7. It was also observed that higher-irradiation exposures caused the loss of renaturation ability ofEscherichia coli DNA.Abbreviations used DNA deoxyribonucleic acid - G guanine - C cytosine - E ₂₆₀ extinction (absorbancy) measured at 260m - T _m the temperature corresponding to the midpoint of the absorbance rise - 2/3 the transition interval of the denaturation curve corresponding to the temperature interval between 17–83% of the total absorbancy increment of the denaturation curve - SSC standard saline citrate buffer (0.15 M NaCl, 0.015 M sodium citrate, pH 7) - PE 10^–2 M sodium phosphate buffer [molarity related to (PO₄)] - PE 10^–3 M EDTA, pH 7. 1.000 ml prepared as follows: 0.608 g NaH₂PO₄.2 H₂O, 0.218 g Na₂HPO₄.12 H₂O, 0.372 g disodium salt of EDTA, 1.0 ml 1 M NaOH. Concentration of Na ions — 0.02 M.     

Absence of α-methyldopamine in rat striatum after chronic administration of d-amphetamine

Direct measurement by gas chromatography methane chemical ionization mass spectrometry of α-methyldopamine and α-methylnorepinephrine in rat striatum has shown the failure of these compounds to be accumulated in vivo after chronic administration of d-amphetamine despite the accumulation of α-methyltyramine, an immediate in vitro precursor. Further, both α-methyldopamine and α-methyltyrosine. These data suggest striatum after administration of α-methyltyrosine. These data suggest that, after administration of α-methyltyrosine, α-methyldopamine is formed via decarboxylation of α-methyldopa and not from hydroxylation of α-methyltyramine. Finally, our results indicate that α-methyldopamine does not play a role in the development of tolerance to d-amphetamine.     

COUMARIN THERAPY—Prothrombin Activity after Termination of Treatment

Twelve patients receiving coumarin type hypoprothrombinemic agents were studied before, during and after termination of therapy, the prothrombin proconvertin method having been used to assay the prothrombin activity complex.In no instance was post treatment “rebound” demonstrated.Prothrombin activity levels returned to pretreatment values only after ten days following termination of coumarin or Dicumarol administration.If a reactivation of thrombotic tendency occurs following discontinuance of anticoagulant therapy, it would not appear to be related to a “rebound” of prothrombin activity above that which is “normal” for the individual patient.Patients tend to return to the same level of prothrombin activity present before initiation of coumarin therapy.     

Frequency of chromosomal aberrations after exposure to γ-radiation of human chorionic villi

We investigated the chromosomal damage induced by in vitro exposure to γ-rays of uncultured first trimester chorionic villi. Frequency and types of chromosomal aberrations at increasing doses of radiation have been evaluated on cytotrophoblast spontaneous metaphases obtained after a short term incubation. Our results indicate a direct correlation between radiation dose and aberration frequency.     

Biosynthetic capacity of hepatocytes after storage at 4°C

Capacity to synthesize glucose, urea, and ketone bodies is well maintained in hepatocytes after storage for at least 24 h at 4 degrees C. Substrates and albumin are the only requirements.     

Localization of lipid peroxidation products in liposomes after γ-irradiation

The distribution of lipid peroxidation products in liposomes after γ-irradiation at various doses was studied. Increases in thiobarbituric-acid-reactive substances, in the absorbance at 232 nm and in hydroperoxides were observed mainly in liposomal membranes after relatively low doses of irradiation, while carbonyl compounds were distributed both inside and outside the membranes. After higher doses of irradiation, however, the absorbance at 232 nm and the amount of hydroperoxides reached a maximal level in the membrane portion and then decreased when the decomposition products were released from the membranes. Under this condition, malondialdehyde and other carbonyl compounds were increased mainly in the medium of liposomal suspension. These results are discussed with reference to the lipid peroxidation process which is induced quantitatively by ionizing radiation.     

Morphinome – proteome of the nervous system after morphine treatment

Summary. Proteome is a natural consequence of the post-genome era when the HUGO project (Human Genome Organization) has almost been completed. Here, a specifically aimed proteome in drug dependence – morphinome, is described, including tasks, strategies and pitfalls of the methodology.     

Relief of pain by infusion of morphine after operation: does tolerance develop?

To see whether continuous intravenous infusion of opiates provides more effective postoperative relief of pain than conventional intramuscular injection these regimens were compared in a prospective double blind trial. Thirty patients undergoing elective cholecystectomy were allocated randomly to receive an infusion of morphine or an infusion of placebo (control group) for 24 hours. Both groups were allowed supplementary morphine boluses as requested. During the first 48 hours after operation the degree of pain was almost identical between the groups. Surprisingly, the group that was given the infusion of morphine received as much supplementary morphine as the control group during the first 24 hours and appreciably more during the 24 hours after the infusion had been withdrawn. Nausea and vomiting were more prevalent among the patients given the infusion of morphine. These results suggest that continuous infusion of morphine may be an inferior regimen to intermittent bolus administration in the relief of postoperative pain. This may be explained by the development of tolerance in patients who received the infusion of morphine.     

Investigate of AQP gene expression in the liver of mice after ischemia–reperfusion

Ischemia–reperfusion (IR) injury usually occurs during liver transplantation. Aquaporins (AQPs) are transmembrane channels that facilitate water permeability through cell membranes and are essential for the regulation of water homeostasis. Changes in the AQPs expression have been correlated with several inflammatory diseases. Less is known about AQPs expression in hepatic ischemia reperfusion injury. To clarify the roles of AQPs in IR injury, in this current study we examined the gene expression patterns of AQP1, 8 and 9 in the liver after IR injury. Male balb/c mice were exposed to partial (70%) hepatic ischemia for 65 min and then randomized into five groups of reperfusion [0 h (A), 8 h (B), 1 day (C), 3 days (D), and 7 days (E)]. A surgical group was also selected as the sham group. Serum and liver tissue samples were collected for evaluation of alanine aminotransferase (ALT), aspartate aminotransferase (AST) and liver histopathology. Real time PCR was performed to evaluate the AQPs expression. I/R injury resulted in a significant increase in ALT and AST (p?<?0.05) compared to sham mice in each group. The gene expression of AQPs was significantly increased in the IR group compared with the sham group (p?<?0.05). AQP8 and AQP1 after 8 h (group B) showed the highest gene expression in comparison with other groups, but the highest level of AQP9 gene expression was observed after 1 day (group C). Pathologic changes in the liver after reperfusion were confirmed the IR. In the IR group cytoplasmic vacuolization, inflammatory cell infiltration and focal necrosis were detected. In conclusion, our findings indicated that the damage caused by ischemia–reperfusion in the liver can change the expression of AQP genes, which can interfere with hepatocellular homeostasis and their function. Upregulation of AQP1, 8 and 9 could contribute to the development of hepatocellular swelling after hepatic IR injury.     

Long-term antidiabetic activity of vanadyl after treatment withdrawal: Restoration of insulin secretion?

In its vanadate (V⁵⁺) or vanadyl (V⁴⁺) forms, vanadium has been demonstrated to possess antidiabetic activity. Oral treatment of streptozotocin (STZ)-diabetic animals with either form is associated with correction of hyperglycemia, and prevention of diabetes-induced complications, although weight gain is unaffected. Vanadium treatment of non-diabetic animals lowers plasma insulin levels by reducing insulin demand, as these animals remain normoglycemic. These results suggest that vanadium hasin vivo insulin-mimetic or insulin-enhancing effects, in agreement with severalin vitro observations.Chronic treatment with vanadium has also been shown to result in sustained antidiabetic effects in STZ-diabetic animals long after treatment has ceased. Thus, at 13 weeks after withdrawal from treatment, corrected animals had normalized glucose and weight gain, and improved basal insulin levels. In addition, near-normal glucose tolerance was found despite an insignificant insulin response. Since vanadium accumulates in several tissue sites (e.g. bone, kidney) when pharmacological doses are administered, it is possible that stored vanadium may be important in maintaining near-normal glucose tolerance at least in the short-term following withdrawal from treatment. Recently, following withdrawal of vanadyl treatment up to 30 weeks, diabetic animals which had remained normoglycemic and had normalized glucose tolerance showed improvements in plasma insulin levels both in the basal state and in response to oral glucose, as compared to those which had reverted to hyperglycemia. The observed significant improvements in insulin capacity over the long-term (>3 months) suggests that a restored and/or preserved insulin secretion may be essential for maintained reversal of the diabetic state over a prolonged period after treatment is withdrawn.     

Isolation of α and β Brain Tubulin Subunits after Alkaline Treatment of the Protein

We demonstrate here that brain purified tubulin can be dissociated into and subunits at pH > 10 and that the subunits can be separated by using the Triton X-114 phase separation system. After phase partition at pH > 10, tubulin but not tubulin behaves as a hydrophobic compound appearing in the detergent rich phase. After three extractions of the alkaline aqueous phase with Triton X-114, about 90% of the tubulin was recovered in the detergent rich phase. The hydrophobic behavior observed for tubulin after its dissociation at pH 11.5 was not due to an irreversible change of the protein, because when the detergent rich phase containing tubulin was diluted with a buffer solution at pH 7.3 and the solution allowed to partition again, -tubulin is recovered in the aqueous phase. The detergent in the aqueous phase of the and tubulin preparations can be removed up to 90% by 12 h dialysis. The and subunits of tubulin from kidney and liver behave, in this phase separation system, like those of brain tubulin.     

Structure of actinidin,after refinement at 1.7 Å resolution

The structure of the sulphydryl protease, actinidin, after refinement at 1.7 Å resolution, is described. The positions of most of the 1666 atoms have been determined with an accuracy better than 0.1 Å; only two residues (219 and 220) and the side-chain of a third (87) cannot be seen. In addition, the model contains 272 solvent molecules, all taken as water, except one which may be an ammonium ion. Atomic B values give a good indication of the mobility of different parts of the structure. Actinidin has a double domain structure, with one domain mostly helical in its secondary structure, and the other domain built around a twisted β-sheet. The geometry of hydrogen bonds in helices, β-structure and turns has been analysed. All are significantly non-linear, with the angle N-?…O ~160 °. Carbonyl groups are tilted outwards from the axis of each helix, the tilting apparently unaffected by whether or not additional hydrogen bonds are made (e.g. to water or side-chain atoms). Each domain is folded round a substantial core of non-polar side-chains, but the interface between domains is mostly polar. Interactions across this interface involve a network of eight buried water molecules, the buried carboxyl and amino groups of Glu35, Glu50, Lys181 and Lys17, other polar side-chains and a few hydrophobic groups. One other internal charged side-chain, that of Glu52, is adjacent to a buried solvent molecule, probably an ammonium ion. Other side-chain environments are described. One proline residue has a cis configuration. The sulphydryl group is oxidized, probably to SO₂^?, with one oxygen atom clearly visible but the other somewhat less certain. The active site geometry is otherwise compatible with the mechanism proposed by Drenth et al. (1975,1976) for papain. The positions of the 272 solvent molecules are described. The best-ordered water molecules are those that are internal (total of 17), in surface pockets, or in the intermolecular contact regions. These generally form three or four hydrogen bonds, two to proton acceptors and one or two to proton donors. Other water molecules make water bridges on the surface, sometimes covering the exposed edges of non-polar groups. Intermolecular contacts involve few protein atoms, but many water molecules.     

β-endorphin levels after intrathecal infusion of iodinated contrast media

Modifications in β-endorphin levels in cerebrospinal fluid have been described following lumbar puncture and metrizamide injection. Cerebrospinal fluid (CSF) samples were obtained from 19 patients before and after lumbar myelography. Two radioimmunoassays were used. One was a commercial kit; and the other one (developed in our laboratory) used a chromatographic removal from β-lipotrophin. No definite variation of β-endorphin was observed after myelography, using either the commercial kit or a more sophisticated procedure. Some controls were prepared by adding metrizamide or Iopamidol in vitro to CSF samples in order to evaluate a non specific effect of these contrast media. The results obtained with these controls suggest that the discrepancy of results may be explained simply by assay artifacts due to drug interferences when using the commercial methods.     

Does the chromophore's ring move after photoexcitation of rhodopsin?

By comparing the shift of the absorption maxima when a visual pigment is converted to its lumirhodopsin photointermediate for two classes of pigments, we can infer whether or not the pigment's beta-ionone ring has left its binding site. We compare this shift for the long-wavelength sensitive visual pigment of chicken iodopsin (lambdamax = 571 nm), which has polar residues in the ring binding site that interact with the ring, with that for three pigments, which do not. We conclude that by the time the Lumi product of the pigment is formed, the ring has moved away from the ring binding site.     

Rate of Synthesis of Non-Ribosomal RNA in Castrate Uterus after Oestradiol-17β Stimulation

OESTROGENS have been reported to stimulate preferentially the synthesis of ribosomal RNA in the castrate uterus^1–4. Thus it has been suggested that 50% or more of the RNA that is synthesized in the oestrogen-stimulated uterus is ribosomal precursor RNA^1,2,4. The concept is supported by the reports of enhanced ribosome formation during early oestrogen action^5,6. It has also been shown, however, that during the first 6 h after oestrogen administration there is no increase in total uterine RNA in the rat uterus^4,7 and also the castrate mouse uterus⁸. These findings seem to be incompatable with the idea that much of the RNA that is synthesized during this first 6 h is ribosomal precursor RNA, most of which accumulates as new stable rRNA. Determination of the absolute rates of total RNA synthesis in vivo should provide some insight into the amounts of various species of RNA that are synthesized after oestrogen administration. Data presented here for the rate of total RNA synthesis strongly suggest that all except a small portion of the RNA that is being synthesized at 4 h after oestrogen stimulation is unstable in vivo and hence is not ribosomal precursor RNA.     

Perturbation of the vesicle cycle in reticulospinal synapses of lamprey after presynaptic microinjections of GTPγS

The synaptic vesicle cycle sustains neurotransmission and keeps exo- and endocytosis in synapses in dynamic equilibrium. GTP-binding proteins function as key regulators of this cycle. The large GTPase dynamin is implicated in the fission of clathrin-coated vesicles from presynaptic membrane during endocytosis. The present study addresses the effect of the nonhydrolysable GTP analog GTPγS on assembly of the dynamin fission complex in situ. Intraaxonal microinjections of GTPγS induced the following distinct ultrastructural changes in the synapses: the number of synaptic vesicles in a cluster decreased while the number of the docked vesicles at the active zone increased; at the same time, the clathrin-coated intermediates also increased in number, indicating the inhibition of synaptic vesicle recycling. Unusual clathrin-coated intermediates were found. At low concentrations of GTPγS, they were presented by long tubules wreathed with a dynamin helix (spiral) and topped with a clathrin-coated vesicle. At high concentrations of GTPγS the tubular structures were much shorter and branched, with each branch topped with a clathrin-coated vesicle. The spiral pitch and the tubule diameter were significantly reduced as the concentration of GTPγS built up (23.1 ± 0.4 and 26.6 ± 0.4 nm, respectively, at low and 19.0 ± 0.5 and 23.3 ± 0.4 nm at high concentration of GTPγS, p < 0.001). We suggest that these ultrastructural changes reflect different steps in dynamin-mediated fission of clathrin-coated vesicles and propose a model for this process. The model implies that at first, GTP hydrolysis leads to a fast elongation of the helix due to a straightening of its dynamin dimmers. This entails an increase both in a pitch and a diameter of the dynamin helix. The shift in diameter disrupts local hydrophobic interactions between the inner and the outer lipid layers of the membrane at the sites of dynamin binding. Concurrent stretching of the helix and the clathrin-coated vesicle’s neck disintegrates the neck membrane and results finally in a release of the clathrincoated vesicle.