Effect of levofloxacine on cell elements of the lung tissue and on the growth of drug resistant Mycobacteria tuberculosis]

Levofloxacin effect on morphokinetic parameters of the murine lung tissue culture was investigated. This model allowed also to evaluate the levofloxacin activity against drug-resistant intracellular and extracellular Mycobacterium tuberculosis: MBC was 0.5-1.0 mg/l. Bactericidal effect started on the 3d day of incubation and was maximal on 7th day. Tissue cells vitality was not changed. Combination of levofloxacin with antimycobacterial drugs of first choice was investigated: isoniazid demonstrated synergistic effect, pyrazinamide also demonstrated synergistic effect. Combination of levofloxacin with rifampicin was antagonistic.     

Review of the in vitro activity and potential clinical efficacy of levofloxacin in the treatment of anaerobic infections

The activity of levofloxacin against aerobic bacteria has been well documented both in vitro and clinically, but its anaerobic activity has been infrequently studied. This new fluoroquinolone exhibits good in vitro activity (MIC(S) < or =2.0 microg/mL) against many anaerobic pathogens associated with acute sinusitis, bite wounds, and other soft-tissue infections. It is less active against Bacteroides fragilis (MIC (90)=2-4 microg/mL ) and has poor inhibitory activity against non-fragilis B. fragilis group species that are associated with gastrointestinal and genitourinary tract infections. Levofloxacin does not antagonize the in vitro activity of clindamycin and metronidazole and often provides additive or synergistic activity against anaerobic bacteria with these agents. In pharmacodynamic models, levofloxacin exhibits rapid bactericidal activity at 2-4 times the MIC of anaerobic bacteria. Prolonged killing is observed when the area-under-the concentration-time-curve to MIC ratio is greater than 40. In clinical efficacy trials, levofloxacin has been effective in the treatment of patients with gynecologic, skin and skin-structure, and bone infections involving anaerobic pathogens. Both micro-biologic and pharmacodynamic studies support further evaluations of levofloxacin in the treatment of selective mixed aerobic/anaerobic infections.     

Bactericidal activity of peripheral blood neutrophils during the oestrous cycle and early pregnancy in the mare

The oestrous cycles of 20 mixed-breed mares were synchronized with daily injections of 10 mg oestradiol-17 beta and 150 mg progesterone given i.m. for 10 days. On the 10th day, 10-15 mg prostaglandin F-2 alpha was administered i.m. to induce oestrus. Neutrophils were isolated from jugular blood on the 2nd or 3rd day of oestrus, Days 5 and 7 after ovulation or during early pregnancy (Days 18-34 of pregnancy). Neutrophils were challenged with Staphylococcus aureus and their bactericidal activity examined after 30 and 120 min of incubation for a reduction of colony forming units. Bactericidal activity increased with the time of incubation (P less than 0.01) but did not differ for the oestrous cycle or pregnancy (P greater than 0.05).     

Resistance to antibiotics and histamine production at the bacteria, isolated from the stomatopharynx of the children with bronchial asthma]

1549 strains of bacteria (gram-positive and gram-negative bacteria and Candida) were isolated from the 117 children (65 with bronchial asthma and 52 healthy one) aged from 3 to 14 years old. Susceptibility of 1213 strains to 20 antibiotics was determined by disk-diffusion method. It was shown that 336 strains produce histidine decarboxylase and for 40 strains the quantity of the produced histamine was measured on the Moeller medium. Histamine-modifying activity was investigated at 32 strains in vitro by fluorometric method. Reduced colonization resistance of the pharynx of the children with bronchial asthma was shown. Spectrum of bacteria with histidine decarboxylase activity was more wide at the children with asthma. Histamine production was about 1 mcg/mL at streptococci, staphylococci and hemophilic bacteria, from 1 to 3 mcg/mL - at bacilli, corynebacteria and Candida, from 1 to 10 mcg/mL - at lactobacilli, enterobacteria, pseudomonades, acinetobacteria. Ability for partial histamine destruction in virowas demonstrated for some strains of lactobacilli, enterobacteria and pseudomonades (initial histamine concentration reduced on 19.4-35.8 per cent after 48-hour incubation).     

The development of the neurons of the glossopharyngeal (IX) and vagal (X) sensory ganglia in chick embryos

The timetable of cell generation, neuronal death and neuron numbers in the fused proximal glossopharyngeal (IX) and vagal (X) ganglion and distal IX and X ganglia were studied in normal and nerve growth factor (NGF) treated chick embryos. 3H-thymidine was injected between the 3rd and 7th days of incubation and embryos sacrificed on the 11th day. Neurons in the distal IX and X ganglia were generated between the 2nd and 5th days of incubation, the peak mitotic activity occurring on the 4th and 3rd days, respectively. Neurons of the proximal IX and X ganglion were generated between the 4th and 7th days, with maximum neuron generation on the 5th day of incubation. Counts of neurons in the 3 ganglia between the 5th and 18th days of incubation showed a maximum of 22,000 on the 8th day in the proximal IX and X ganglion and this decreased to 12,000 by the 13th day. In the distal IX ganglion, the neuron number decreased by 44% from 4,500 on the 6th day to 2,500 by the 11th day. A similar decrease of 43% was found in the distal X ganglion, the neuron number falling from 11,500 on the 7th day to 6,500 by the 11th day of incubation. Neuronal cell death in these ganglia extended from the 5th to the 12th day of incubation, maximum cell death occurring at or after the cessation of mitotic activity. NGF administration from the 5th to the 11th day of incubation did not have a measurable effect on the neurons of proximal IX and X and distal IX ganglia, but increased neuronal survival by 30% in the distal X ganglion.(ABSTRACT TRUNCATED AT 250 WORDS)     

Circadian rhythms in the incidence of apoptotic cells and number of clonogenic cells in intestinal crypts after radiation using normal and reversed light conditions

Variations in the number of radiation-induced morphologically dead or dying cells (apoptotic cells) in the crypts in the small intestine of the mouse have been studied throughout a 24-h period under a normal light regimen (light on, 07.00-19.00 h; light off, 19.00-07.00 h). A clear circadian rhythm was displayed in the apoptotic incidence 3 or 6 h after irradiation for each gamma-ray dose studied (range 0.14-9.0 Gy). The most prominent circadian rhythm was obtained after 0.5 Gy. The peak time of day for inducing apoptosis was 06.00-09.00 h, and the trough occurred at 18.00-21.00 h. Some mice were also transferred to a room with the light cycle reversed, and were irradiated on different days after the transfer. The apoptosis induced by 0.5 Gy or 9.0 Gy, or the number of surviving crypts (microcolonies) after 11.0 Gy or 13.0 Gy was examined. The transition point for reversal (i.e. the switch time from the normal-light pattern to the reversed-light pattern) of the circadian rhythm in apoptosis (after 0.5 Gy) occurred 7 days after the transfer and the rhythm was reversed by 14 days. The rhythm for crypt survival (i.e. for clonogenic cell radiosensitivity) was disturbed on 1 day and the transition point for reversal occurred 3 days after the transfer. The rhythm became reversed by 7 days. These observations are discussed in relation to the identity of clonogenic cells, (functional) stem cells, proliferating transit cells and the cells sensitive to small doses of radiation (i.e. hypersensitive cells) in the crypt.     

Disorders in the development of the crystalline lens in chick embryos following exposure to insulin]

Inection of insulin into the chicken egg-white in dosage of 4 i. u. during the 1st - 14th days of incubation results in degenerative changes in the lens. Injection of insulin during the 1st day of incubation (before the beginning of eye differentiation) fails to prevent development of the eye vesicle and determination of the lens. The morphogenesis of the lens goes normally up to the 10-13th days of incubation. From this moment abrupt degenerative alterations make their appearance as well as destruction of the lens fibres. When the lens is affected by insulin the destruction is observed on the following day. The mechanism of insulin effects upon the lens seems to be as follows: Exogeneous insulin remains in the embryo tissues for along time. When the tissues are preparing to accept endogeneous insulin and become susceptible to it (the 10th-11th days of incubation) exogeneous insulin begins its pathological influence on the lens fibres. After the 13th-14th days of incubation endogeneous insulin comes into the blood. Against the background of exogeneous insulin it results in excessive increase of concentration of insulin in the embryo tissues and great degenerative changes in the lens.     

Precocious induction of luteal activation and termination of delayed implantation in mink with the dopamine antagonist pimozide

The effects of the dopamine antagonist pimozide on the preimplantation delay phase of mink gestation were investigated in field and laboratory trials. Three doses of 0.1 mg pimozide in acetic acid administered on the 7th, 9th and 11th days after mating abbreviated gestation in Pastel kit female mink to a mean (+/- SEM) of 45.5 +/- 0.5 days, 10 days less than that observed in mink treated with vehicle only (55.6 +/- 0.6 days). In laboratory trials, four doses of 0.1 mg pimozide on the 7th, 9th, 11th and 13th day after mating resulted in embryo implantation at a mean of 25 +/- 4.3 days after mating while vehicle-treated control animals had mean preimplantation delay of 37 +/- 3.1 days. Luteal activation in the pimozide-treated group, as indicated by a rapid increase in circulating progesterone, began within 2 days after the first pimozide injection. No increase was observed in vehicle-treated mink until 6 or more days after the initiation of injections or 13 days after mating. It was concluded that pimozide, presumably by permitting endogenous secretion of prolactin, can induce precocious luteal activation and embryo implantation in the mink.     

Complex of recombinant heat shock protein with lipopolysaccharide induces rapid protection of mice against Salmonella typhimurium and avirulent for humans avian influenza virus H5N2

Protective effect of immunization with heat shock protein (HSP) against bacterial and viral infections in mice was studied. Recombinant HSP 70 kDa of Mycobacterium tuberculosis contaminated with lypopolysaccharide (0.185 mcg/ml) was used for experiments. One intraperitoneal injection of 100 or 400 mcg of HSP induced rapid protection against intraperitoneal challen e with 125 LD50 of Salmonella typhimurium (on 3rd-6th day) and against intranasal challenge with 10 LD50 of avirulent for humans avian influenza virus H5N2 (A/ mallard/Pennsylvania/10218/84) (on 5th-8th day). Three daily injections with 10 mcg of HSP induced rapid, significant and long-term protection against S. typhimurium. Immunization with HSP protected 100% of mice during 3 days after the challenge, 50% of immunized animals survived during 21 days (duration of the study). All nonimmunized mice died on 6th day.     

The influence of biosynthetic human growth hormone on biomechanical properties and collagen formation in granulation tissue

Biomechanical properties and collagen formation in the granulation tissue of cellulose sponges, implanted subcutaneously in male rats for 7, 10 and 16 days, were tested after treatment with biosynthetic human growth hormone given subcutaneously in a dose of 0.5 mg/kg body weight/day. At each implantation period, one group started hormone treatment at the day of implantation and another group started hormone treatment 7 days prior to implantation. After 7 days of implantation, increases in maximum stress (36 per cent), relative failure energy (48 per cent) and strain at maximum stress (25 per cent) were found when treatment was started 7 days prior to implantation. After 10 days of implantation an increase in relative failure energy (60 per cent) was found when treatment was started 7 days prior to implantation. No differences were found after 7 and 10 days of implantation when treatment was started at the day of implantation. After 16 days of implantation, no influence on mechanical strength was found in any of the hormone treated groups. The collagen deposition after 7, 10 and 16 days did not differ in any of the hormone treated groups compared to controls.     

Ecdysterone and an analogue of juvenile hormone on the autophagy in the cells of fat body of mamestra brassicae.

The effect of ecdysterone and a juvenile hormone analogue (JHa) on autophagy and heterophagy was investigated in the fat body cells of the last larval instar of Mamestra brassicae. In the course of normal development autophagic vacuoles and protein granules of heterophagic origin begin to accumulate in these cells, on the 4th and 5th day of the last larval stage respectively. When ecdysterone (10 mug/g body weight) was administered to the larvae for 24 h either on the 1st or on the 2nd day of the last larval stage, autophagic and heterophagic vacuoles appeared in the cells as early as on the 2nd or 3rd days. Autophagy was also observed in the cells of one-or two-day-old last larval fat body after a 5 h incubation in a medium containing 10 mug/ml ecdysterone, in vitro. Ligation of the last thoracic segment resulted in inhibition of metamorphic changes in the fat body lobules of the isolated abdomen. Injection of 10 mug ecdysterone into the isolated abdomen resulted in an appearence of autophagic vacuoles in these cells, too. JHa treatment, when started on the 2nd or 3rd day of the last larval stage, inhibited both auto- and heterophagy and the fat bodies maintained their larval character. Treatment started on the 4th or 5th day proved either ineffective or lethal. It is concluded that the auto- and heterophagy taking place in the larval fat body cells are stimulated by ecdysterone and inhibited by JHa. Experiments performed in vitro or on ligated animals in vivo provided evidence for a direct action of ecdysterone at the cellular level.     

Detection of viable, but non-culturable Pseudomonas fluorescens DF57 in soil using a microcolony epifluorescence technique

Abstract Kanamycin-resistant Pseudomonas fluorescens DF57-3 cells (Tn5 modified) inoculated in soil microcosms rapidly lost their culturability, as defined by visible colony formation on Kings B agar supplemented with kanamycin. Thus, after 40 days only 0.02–0.35% of the initial inoculum was culturable. A microcolony epifluorescence technique was developed to determine the viable, but non-culturable subpopulation. A suspension of bacteria from the soil was prepared in salt solution after a sonication procedure and a sample was filtered onto a 0.2 μm Nuclepore filter. The filter was then placed for 3–4 days on the surface of Kings B agar before staining with acridine orange for epifluorescence microscopy. By staining and washing the filters carefully, disruption of microcolonies could be avoided. A majority of the microcolonies resulted from 2–3 cell divisions during the first 2 days of the incubation period, after which the cell divisions stopped. These microcolonies were taken to represent a population of viable, but non-culturable cells and comprised about 20% of the initial inoculum. A similar recovery was obtained when the filters were incubated on the surface of citrate minimal medium or soil extract medium. A few microcolonies showed continued growth on the filters, however, and their number corresponded well with that of visible macrocolonies. Observation by microscopy of a few (2–3) cell divisions (microcolony epifluorescence technique) is proposed for determination of subpopulations of viable, but non-culturable bacteria in soil.     

Effects of thyroxine on mitochondrial morphology and uptake of 35S in embryonic chicken livers

Ultrastructural analyses of reactions of mitochondria in hepatocytes of chicken embryos to low levels of exogenous thyroxine (T4) reveal that such reactions (overall swelling accompanied by disruption of crest geometry) first take place at about 10 days of incubation, T4 having been administered on the 6th day. Physically altered mitochondria may be seen after 11-12 days of incubation but are no longer evident by 13 days. Correlated with the initial evidence of T4 effects on mitochondria at 10 days of incubation is a spurt in hepatocyte proliferation. The time correlation observed between T4 induced mitochondrial changes in morphology and abrupt increases in rates of cell proliferation, suggests that liver nuclear receptors for thyroxine are unavailable prior to 9-10 days of incubation. Golgi complexes within the hepatocytes appear to be especially active in the production of electron-opaque vesicles from at least the 8th day of incubation to 11-12 days. Uptake of 35S (probably into chondroitin sulphates) was found to be fifteen times greater on the 8th day of incubation than at 15 days. This correlates with the period of heightened activity of the Golgi complex. In livers exposed to T4 on the 6th day, uptake of 35S was higher on the 9th and 10th days of incubation as compared to controls.     

Effects of exposure time using preliminary concentrations on the mutagenic effect of the basic action of one-center and dual-center mutagens

The effect of exposition with pretreatment for thiophosphamide and dipin of human lymphocytes at Go phase was investigated. There were used 5 low concentrations of mutagens: 2, 0, 2; 2.10(-2); 2.10(-3), 2.10(-4) mcg/ml with different exposure: 1/4 hr, 1/2 hr, 1 hr and 4 hr and high concentration of 20 mcg/ml by which cell have been treated. There was discovered the dependence of the "protective" concentration on the exposition: the increase of exposition of pretreatment induced the decrease of "protective" concentration and vice versa.     

Effect of dimethyl sulfoxide on transformed rat Schwann cells

Cultured rat Schwann cells transformed by Simian Virus 40 (SV40) have previously been shown to retain their ability to synthesize myelin-associated galactosylceramide and sulfatide. Little is known about the mechanism regulating galactosphingolipid synthesis in Schwann cells. We have found that growing the transformed Schwann cells in the presence of dimethyl sulfoxide (DMSO) markedly inhibits the incorporation of [35S]sulfate into sulfatide, in a time- and dose-dependent manner. The concentration of DMSO which resulted in a half-maximal inhibition after 6 days of incubation was 0.5%, and the incubation time required for a half-maximal effect at 1.0% DMSO was approximately 4 days. In contrast, DMSC did not affect the incorporation of [35S]sulfate into glycosaminoglycans. In addition, DMSO treatment has little effect on the synthesis of cellular DNA, proteins and lipids. When transformed Schwann cells were treated with DMSO, a substantial decrease in the incorporation of [3H]galactose into galactosylceramide was observed. The concentration of DMSO which resulted in a half-maximal inhibition of galactosylceramide synthesis was approximately 0.5%, similar to the concentration required for a similar effect on sulfatide synthesis. However, the incubation time required for a half-maximal inhibitory effect on galactosylceramide synthesis at 1.0% DMSO was less than 1 day, which was substantially shorter than the time required for the inhibition of sulfatide synthesis at this concentration. This finding is consistent with the interpretation that treatment with DMSO inhibits the synthesis of galactosylceramide, a precursor of sulfatide, which results in a decrease in the synthesis of sulfatide during a prolonged incubation of DMSO.     

Are spontaneous activity and supraspinal control co-responsible for functional development of the generator of embryonic motility?

The effect of chronic administration of phenobarbital (from the 4th to the 16th day of incubation) combined with chronic spinal decentralization (decapitation at stage 11-13) on the development of spontaneous motor activity (recorded on the 17th day of incubation) was studied in 4 to 17-day-old chick embryos. 1. Combination of the two experimental treatments led to summation of their negative effect on development; spontaneous motor activity fell to 15.2% of the control value (to 26.9% after the isolated administration of phenobarbital and to 30.5% after isolated decapitation). 2. Metrazol activation of motor activity (100 mg/kg egg weight) after combination of the two factors was relatively no different from the results after their isolated use. 3. The acute administration of GABA (100 mg/kg e.w.) likewise induced relatively the same depression of spontaneous motility as after isolated chronic decapitation and the isolated chronic administration of phenobarbital. The results confirm the hypothesis of the significance of spontaneous activity of the spinal motoneurones for their survival and their actual functional development during the embryonic period.     

黄瓜去顶苗直接成花的形态学研究

本文对黄瓜0—7天幼苗及去顶后0—8天诱导花芽分化苗与诱导营养芽分化苗的子叶节进行了系统石蜡切片观察,未发现0—7天幼苗的子叶叶腋存在潜伏芽。去顶后1—2天在子叶叶柄基部与切口之间的表皮下细胞分裂形成突起,去顶后6天诱花苗与诱芽苗的突起表现出形态差异,诱花苗突起的上端变钝,而诱芽苗突起的上端成尖锥状。去顶后8天诱花苗在子叶叶柄与切口之间形成完整的花芽。另发现有少量花芽起源于切口处细胞。对去顶后0—6天诱花苗与诱芽苗的子叶节还进行了电镜扫描观察,观察结果与石蜡切片基本一致。     

The acute and the chronic effects of imipramine on the spontaneous motility in chick embryos

The effect of imipramine on the spontaneous motility and development of chick embryos was studied from the 4th to the 19th day of incubation. On acute administration (a single dose of 12.5 of 25 mg/kg egg weight), imipramine already induced significant depression of spontaneous motility in 11-day embryos--an effect which increased significantly after the 15th day of incubation. The similar effect of imipramine in spinal embryos testifies to its direct action on the spinal cord and draws attention to certain details of the role of supraspinal structures of the CNS in the acute effect of imipramine. The chronic administration of imipramine showed that it had an almost 100% lethal effect from 4th to the 7th day of incubation. Between the 8th and the 10th day it caused longlasting depression of spontaneous motility. When it was administered between the 11th and 16th day of incubation, no significant effect on the development of spontaneous motor activity was found in chick embryos.     

Feline ocular chlamydiosis: clinical and microbiological effects of topical and systemic therapy

Conjunctival swabs taken from a two-month-old kitten showing ocular discharge were found to be positive for Chlamydophila felis by PCR and isolation. The cat was treated with topical 1% tetracycline ophthalmic ointment twice a day for 60 days. At the end of the treatment, the cat showed no ocular signs and conjunctival swabs resulted PCR and isolation negative for C. felis. Forty days later, the ocular discharge recurred and C. felis was isolated from conjunctival swabs taken from both the cat's eyes. Twenty days of doxycycline systemic treatment at 10 mg/kg once daily was started. The treatment resulted in a complete clinical recovery after a few days. C. felis was not isolated or amplified on the 10th day after beginning the treatment. The cat's conjunctival swabs were also PCR and isolation negative on the 10th, 30th, 60th, 90th, 120th and 240th days after the end of therapy.     

蛋白激酶与细胞分化—两种分化诱导剂对酪氨酸蛋白激酶的抑制

在报道视黄酸(RA)是人肝癌细胞株SMMC-7721分化诱导剂的基础上,本文继续报道8-溴-环磷酸腺苷对该细胞也有分化诱导作用,两者都抑制细胞的增殖,降低γ-谷氨酰转肽酶(γ-GT)比活力和升高(ALP)碱性磷酸酶的比活力。在10μmol/LRA和0.5mmol/L-8-Br-cAMP处理细胞1、3、5天后,胞液和膜性组分中的酪氨酸蛋白激酶(TPK)的比活力均降低,其中RA对胞液TPK的作用在早期较明显,约降低30％,而对膜性TPK的影响则随培养天数而逐渐增加,至第5天下降达50％以上。8Br-cAMP则相反,对胞浆TPK的抑制主要发生在3天以后;约抑制43—53％,而对膜性组分则抑制率逐日降低,在第一天较为明显。因TPK是一个细胞增殖恶变的标志,故RA和8-溴-cAMP对TPK的抑制进一步证明这两种分化诱导剂对SMMC-7721细胞的逆转作用。