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1.
目的选择一种最优势、合理的固定方式以提高对睾丸组织制片效果,以配合不同种类的科学研究。方法选用10%甲醛溶液、NBF—Bouin’s、Bouin’s和改良Davidson’s四种不同的固定液对大鼠睾丸进行充分固定后,制作石蜡切片,进行HE染色,比较不同固定液中的睾丸组织学形态的差异;利用糖原特殊染色(PAS),探讨不同固定液对睾丸糖原观察的影响;采用免疫组织化染色,测评睾丸组织内雄激素受体的固定效果。结果改良Davidson’8固定液较NBF—Bouin’s引起的曲细精管萎缩轻,形态更为清晰,用免疫组织化学方法检测雄激素更为敏感,并且改良Davidson's固定液在需要对精子发生进行分期时,其PAS染色的效果与Bouin's液固定后等同。结论与苴守圈常浦相№曲冉David0Rnn浦对女宙奥由的圈索特罩掂杯  相似文献   

2.
The effect of preservation methods on dry weight (DW) and ash-free dry weight (AFDW) of Radix peregra (Gastropoda), Asellus aquaticus (Isopoda), Erpobdella octoculata (Hirudinea) and Glyptotendipes sp. (Chironomidae) was studied. Ethanol, formaldehyde, and Bouin were used as preservative. In case of preservation of macro-invertebrates in ethanol substancial changes in DW and AFDW biomass were observed. In the four different taxa the loss in DW varied between 7.2–21.9% after a 3 month preservation period in 70% ethanol. A comparatively small range in AFDW loss (16.2–19.7%) was found. Changes in DW and AFDW biomass during preservation were significantly affected by the duration of the preservation, by temperature, light conditions and the volume of the preservative. The changes in AFDW were also significantly affected by the concentration of the preservatives. Preservation in 10% formaldehyde did not cause significant changes in DW and AFDW biomass.Contribution nr. 42 of the nymphalid project.  相似文献   

3.
Olive trees have a plentiful bloom but a low percentage of normal fruit set. To improve fruit set, numerous investigations have sought to identify the obstacles that prevent full production. In this work, flower development in five DOP Umbria cultivars (Leccino, Frantoio, Moraiolo, Dolce Agogia and San Felice) was studied throughout different developmental phases, from before microsporogenesis and megasporogenesis to post-anthesis, by morphological and cyto-histological observations. Dolce Agogia was the most precocious cultivar, while full flowering was simultaneous in Leccino, Frantoio, Moraiolo and San Felice. Frantoio and Leccino were also good pollen producers, having the highest percentage of pollen viability and germinability. Dolce Agogia can also be considered a good pollen producer in terms of the high quantity of released pollen, but it had the lowest levels of pollen viability and germinability and the highest percentage of aborted flowers and ovaries. Morphological and cyto-histological observations on the number of flowers per inflorescence and the number of aborted flowers and ovaries suggest that fruit set was not influenced by the number of flowers per inflorescence, but rather by the number of inflorescences, which depends on the global fruiting potential of the tree.  相似文献   

4.
Paecilomyces variotii IRI017 was isolated as a formaldehyde-resistant fungus from wastewater containing formaldehyde. The fungus grew in a medium containing 0.5% formaldehyde and had consumed formaldehyde completely after 5 days. Alcohol oxidase was purified from the fungus grown on methanol. A 20-fold purification was achieved with a yield of 44%. The molecular mass of the purified enzyme was estimated to be 73 and 450 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and gel filtration chromatography, respectively, suggesting that the enzyme consists of six identical subunits. The N-terminal amino acid sequence of the subunit was TIPDEVDIII. The enzyme showed an absorption spectrum typical of a flavoprotein and had a noncovalently bound flavin different from FAD, FMN, and riboflavin. The pH optimum of the enzyme activity was pH 6–10. The enzyme was stable in the pH range of pH 5–10. The enzyme retained full activity after incubation at 50°C for 30 min. The enzyme oxidized not only methanol but also lower primary alcohols and formaldehyde. The K m values for methanol, ethanol, and formaldehyde were 1.9, 3.8, and 4.9 mmol l−1, respectively.  相似文献   

5.
The presence and localization of nerve growth factor receptors (NGFr) in the choroid plexus of the adult rat has been investigated immunohistochemically using an anti-rat NGFr monoclonal antibody (192-IgG). A moderate to strong immunoreaction was observed in the epithelial cells of the choroid plexus, whereas the choroidal blood vessels and connective tissue remained unlabelled. Moreover, no sex-differences were encountered in the NGFr immunoreaction intensity and Bouin fixative was more effective than 10% formaldehyde evidenciating the NGFr immunostain. Occasionally, ependymal cells displaying NGFr immunoreactivity were observed. Present data demonstrate that the choroid plexus of the rat contain NGFr, probably low-affinity NGFr, and suggest an involvement of NGF in the regulation of cerebrospinal fluid secretion, but the importance of these findings, if any, must be investigated in future studies.  相似文献   

6.
Formaldehyde is used routinely in the hot-water treatment (HWT) of narcissus bulbs for the control of stem nematode (Ditylenchus dipsaci) and basal rot (caused by Fusarium oxysporum f.sp. narcissi). Formaldehyde is unpleasant for operators to use, and does not kill all of the thick-walled chlamydospores of F. oxysporum and so less hazardous but more effective materials are being sought. Peroxyacetic acid (as Jet 5, a commercial disinfectant containing 5% peroxyacetic acid) was evaluated in vitro and in a field trial as a possible alternative to formaldehyde. In laboratory studies, peroxyacetic acid (as 1% Jet 5) was as effective as formaldehyde (as 0.5% commercial formalin containing 38 to 40% formaldehyde) in killing free-swimming stem nematodes and nematodes in the wool stage. Peroxyacetic acid (as 0.5% Jet 5) killed F. oxysporum chlamydospores within 1 h, whereas total kill was not achieved with formaldehyde (concentration as above) after 4 h. In a 2 year field trial, there was no evidence of detrimental effects on a healthy narcissus stock due to using peroxyacetic acid. In an infested, diseased stock, bulbs were virtually destroyed by stem nematode within 2 years when HWT was not given. The greatest reduction in nematode symptoms, and the highest bulb yields, were found when formaldehyde or the higher rates of peroxyacetic acid were used in combination with thiabendazole.  相似文献   

7.
The solvent-tolerant bacterium Pseudomonas putida S12 was engineered to efficiently utilize the C1 compounds methanol and formaldehyde as auxiliary substrate. The hps and phi genes of Bacillus brevis, encoding two key steps of the ribulose monophosphate (RuMP) pathway, were introduced to construct a pathway for the metabolism of the toxic methanol oxidation intermediate formaldehyde. This approach resulted in a remarkably increased biomass yield on the primary substrate glucose when cultured in C-limited chemostats fed with a mixture of glucose and formaldehyde. With increasing relative formaldehyde feed concentrations, the biomass yield increased from 35% (C-mol biomass/C-mol glucose) without formaldehyde to 91% at 60% relative formaldehyde concentration. The RuMP-pathway expressing strain was also capable of growing to higher relative formaldehyde concentrations than the control strain. The presence of an endogenous methanol oxidizing enzyme activity in P. putida S12 allowed the replacement of formaldehyde with the less toxic methanol, resulting in an 84% (C-mol/C-mol) biomass yield. Thus, by introducing two enzymes of the RuMP pathway, co-utilization of the cheap and renewable substrate methanol was achieved, making an important contribution to the efficient use of P. putida S12 as a bioconversion platform host.  相似文献   

8.
Neurons of Aplysia californica are naturally pigmented and the pigment accumulates with age. In the present study the pigment was examined in the same neuron from Aplysia of three postmetamorphic ages: young, sexually mature, and old. The large central neuron, R2, was examined by light and electron microscopy to determine if the pigment possessed properties similar to lipofuscin pigment seen in aging mammalian neurons. We used the same microscopic techniques that demonstrate the presence of lipofuscin in mammalian neurons. Light microscopic studies demonstrated a regional correlation between autofluorescence, staining with Sudan Black, and the naturally occurring pigment in old R2s. Electron microscopic studies revealed the presence of large vacuolated and lamellated membrane-bound bodies in the peripheral cytoplasm of old R2s, similar to those found in mammalian neurons. The bodies were located in the same region in which autofluorescence and Sudan Black staining were observed. Although the naturally occurring pigment accumulates with age, it acquires characteristics of lipofuscin pigment in the neurons of older sexually mature animals. The presence of these pigment characteristics can be used as an index of aging in Aplysia neurons as they are in mammalian neurons.  相似文献   

9.
Summary Four strains ofPseudomonas putida and two unidentifiedPseudomonas species that were resistant to hexahydro-1,3,5-triethyl-s-triazine (HHTT) were shown to be resistant to formaldehyde as well. Conjugation experiments revealed that: (a) HHTT and formaldehyde resistance was cotransferred in every case where exconjugants were recovered; (b) in every case HHTT resistance and formaldehyde resistance were expressed to the same level in the exconjugant as in the donor; (c) resistance to either HHTT or formaldehyde alone was never observed; and (d) in instances where HHTT and formaldehyde resistance in the exconjugants was unstable, the exconjugants lost resistance to both agents simultaneously and never to one agent alone. Resistant organisms (e.g.P. putida 3-T-152) had high levels of formaldehyde dehydrogenase and this enzyme appeared to be constitutively expressed. It was concluded that resistance to HHTT was due to resistance to its degradation product, formaldehyde, via detoxification of formaldehyde by formaldehyde dehydrogenase. HHTT- and formaldehyde-sensitive organisms had barely detectable levels (most likely repressed levels) of formaldehyde dehydrogenase. Although speculative, it is possible that formaldehyde resistance may be due to a mutation resulting in derepression of the gene coding for formaldehyde dehydrogenase. While it could not be discerned whether HHTT resistance and formaldehyde resistance were carried on two separate but closely linked genes or if only one gene was involved, the evidence suggested that only one gene was involved. Similarly, it could not be determined whether HHTT and formaldehyde resistance was encoded by chromosomal or plasmid genes.  相似文献   

10.
Abstract Paraffin sections from animal or human tissues fixed in different fixatives were submitted to immunostaining with the mouse monoclonal antibody 19A2, developed by Ogata et al. (1987a) against cyclin/PCNA. Detection of the bound antibody was performed by the indirect method with biotinylated sheep antibody and streptavidin-biotin-peroxidase complexes. No, or faint, nuclear staining was seen in material fixed in ethanol, Bouin, Bouin-Hollande, Carnoy or formaldehyde, whereas readily detectable immunocytochemical reaction was constantly observed over nuclei of methanol-fixed tissues. Hydrolysis with 2 N HCl prior to immunocytochemistry (as currently performed to render incorporated BrdU accessible to antibodies) somewhat improved the results with Bouin or Carnoy and markedly augmented the intensity of the peroxidase reactions in formaldehyde and in methanol-fixed tissues. The distribution of the positive nuclei in the two latter cases coincided with the proliferative compartment. On the other hand, double labelling with [3H]-thymidine and with the cyclin/PCNA antibody revealed that in methanol-fixed tissues the cyclin/PCNA labelling index did not differ by more than 6% from the [3H]-thymidine index. Besides the two labels overlapped in a proportion of labelled cells that was in reasonable agreement with expectation considering cells flow in and out of S phase since the time of [3H]-thymidine injection. This indicates that both labels recognize the same cells in this material. In contrast, in formaldehyde-fixed tissues, the cyclin/PCNA labelling index markedly exceeded the [3H]-thymidine labelling index. From this it is concluded that cyclin/PCNA immunostaining can be used:
  • 1 In formaldehyde-fixed tissues (including existing material stored as paraffin blocks): for defining and mapping the proliferative (or germinative) compartment.
  • 2 In methanol-fixed tissues as a substitute to the [3H]-thymidine autoradiographic labelling index.
From this, a method is proposed (derived from classical ‘double-labelling’technique) for measuring S phase duration in tissues fixed at a known interval time after a single labelling with [3H]-thymidine (or BrdU) and submitted to cyclin/PCNA immunocytochemical detection and to autoradiography (or to BrdU immunostaining).  相似文献   

11.
Pseudomonas putida F61-a defective in formaldehyde dehydrogenase was derived from the parent strain (F61). The bacterial strain grown on a nutrient broth supplemented with 1% glucose exhibited high formaldehyde dismutase activity. The dismutation and cross-dismutation of aldehydes occurred stoichiometrically in the resting-cells reaction. Many kinds of aliphatic and aromatic aldehydes that are scarcely soluble in water were utilized in these reactions as well as soluble ones. Formaldehyde at an extremely high concentration (0.5 M) was almost completely converted to equimolar amounts of methanol and formic acid by the resting-cells, which could be used three times without a loss of activity. The cross-dismutation between acrolein and formaldehyde occurred efficiently in the resting-cells reaction with 0.1 M each substrate. The alcohol: aldehyde oxidoreduction of the short-chain substrates was also shown by the resting-cells of a mutant (M6) unable to grow on n-propanol.  相似文献   

12.
【背景】甲基营养菌(Methylobacterium)是一类能够以单碳或非C-C键低碳化合物(如甲烷、甲醇、甲醛等)为底物生长,并可生产多种代谢产物如氨基酸、工业酶和辅助因子、多羟基烷酸酯(polyhydroxyalkanoates,PHA)、多糖和类胡萝卜素等的革兰氏阴性细菌。【目的】通过突变甲基营养菌MB200的mutS基因,在胁迫条件下定向诱导,以获得可以耐受高浓度甲醇和甲醛的生产菌株。【方法】利用三亲本结合构建mutS基因缺失的高突变菌株MB200sTB,逐步提升培养液中甲醇、甲醛的浓度进行定向诱导突变,对获得的高耐受性突变株进行回补,分析菌株的生长情况。【结果】构建了mutS基因的缺失突变体MB200sTB,并且得到了高耐受甲醇和甲醛的菌株MB200sHBc和MB200sHBq。MB200sHBc与野生株MB200相比其甲醇耐受性得到了极显著的提高,甲醇耐受浓度从8g/L提升到44g/L,但生长量不受影响。MB200sHBq在以甲醛为0.45g/L的碳源条件下,生长量相较于野生型MB200提高了1.69倍。【结论】通过定向诱导缺失mutS基因的突变体,可获得具有生产应用潜力的...  相似文献   

13.
The influence on the area and numerical density of nuclei was investigated in 5-mm-thick slices of guinea pig liver for different fixatives and variations in tissue processing: delay in fixation, air drying, degree of acidity of 10% formalin (= 4% formaldehyde), Bouin and mercury-formalin fixatives, acetone and ethanol dehydration and understretching and overstretching of the paraffin-embedded sections. Air drying (either forced or as a result of delayed fixation), the type of fixative and the degree of acidity affected the nuclear area. Regarding the latter, nuclear area was approximately 25% lower for pH less than or equal to 3 as compared with pH greater than 5. In comparison with the standard tissue processing used, the nuclear density was higher after all of the variations studied (air drying, acetone dehydration and fixation). These findings indicate that nuclear area, in contrast to other tissue components, is relatively insensitive to variations in tissue processing. However, it is essential to regularly measure the pH of the fixative: deviations from pH = 7 should be carefully avoided in order to keep nuclear area variations as a result of tissue processing within acceptable limits.  相似文献   

14.
Summary The mechanism of antimicrobial action of hexahydro-1,3,5-triethyl-s-triazine (HHTT) was studied using the HHTT-resistant isolate,Pseudomonas putida 3-T-152, its HHTT-sensitive, novobiocin-cured derivative,P. putida 3-T-152 11:21,P. putida ATCC 12633,Pseudomonas aeruginosa PA01 andEscherichia coli J53 (RP4). HHTT was oxidized byP. putida 3-T-152, while respiration ofP. putida 3-T-152 11:21 was inhibited by HHTT. Chemical assays showed that HHTT released formaldehyde.P. putida 3-T-152 was highly resistant to formaldehyde, whileP. putida 3-T-152 11:21 was highly sensitive to formaldehyde. Both HHTT and formaldehyde acted similarly to inhibit proline uptake in bacterial cells and to inhibit the synthesis of the inducible enzymes, -galactosidase and glucose-6-phosphate dehydrogenase. HHTT did not have uncoupler-like activity.P. putida 3-T-152 used either HHTT or ethylamine, a component of HHTT, as a nitrogen source for growth, but neither HHTT, ethylamine or formaldehyde served as a carbon and energy source for growth. We concluded that a major mechanism of antimicrobial action of HHTT was through its degradation product, formaldehyde.  相似文献   

15.
Degradation of microbiodies in the methanolutilizing yeastCandida boidinii was mainly studies by electron microscopical observation. The yeast cells precultured on methanol medium contained five to six microbodies per section and showed high activities of alcohol oxidase, catalase, formaldehyde dehydrogenase and formate dehydrogenase. When the precultured cells were transferred into an ethanol medium the number of microbodies and concomitantly the activities of alcohol oxidase and catalase decreased. After 6 h of cultivation microbodies were hardly detected. Also the activity of alcohol oxidase was not measurable and catalase activity was reduced to one tenth, whereas the activities of formaldehyde dehydrogenase and formate dehydrogenase decreased only to about 70%. Experiments with methanol-grown cells transferred into an ethanol medium without nitrogen source indicated that the inactivation of alcohol oxidase and catalase does not require protein synthesis. However, the reappearance of these enzymes is presumably due to de novo protein synthesis as shown by experiments with cycloheximide.  相似文献   

16.
Incubations of Arthrobacter P1 in batch culture in media with mixtures of acetate and methylamine resulted in sequential utilization of the two carbon substrates, but not in diauxic growth. Irrespective of the way cells were pregrown, acetate was the preferred substrate and subsequent studies showed that this is due to the fact that acetate is a strong inhibitor of the methylamine transport system and amine oxidase in Arthrobacter P1. An analysis of enzyme activities in cell-free extracts showed that synthesis of amine oxidase occurred already in the first growth phase with acetate, whereas rapid synthesis of hexulose phosphate synthase was only observed once methylamine utilization started. It is therefore concluded that in Arthrobacter P1 the synthesis of the enzymes specific for methylamine oxidation is not regulated co-ordinately with those involved in formaldehyde fixation, but induced sequentially by methylamine and formaldehyde, respectively.During growth of Arthrobacter P1 on the same mixture in carbon- and energy source-limited continuous cultures both substrates were used simultaneously and completely at dilution rates below the max on either of these substrates. Addition of methylamine, in concentrations as low as 0.5 mM, to the medium reservoir of an acetate-limited continuous culture (D=0.10 h-1) already resulted in synthesis of both amine oxidase and hexulose phosphate synthase. In the reverse experiment, addition of acetate to the medium reservoir of a methylamine-limited continuous culture (D=0.10 h-1), acetate was initially only used as an energy source. Synthesis of the glyoxylate cycle enzymes, however, did occur at acetate concentration in the feed above 7.5–10 mM. This indicates that at acetate concentrations below 10 mM the metabolism of the C1 substrate methylamine is able to cause a complete repression of the synthesis of the enzymes involved in carbon assimilation from the C2 substrate acetate.Abbreviations HPS Hexulose phosphate synthase - MS mineral salts - RuMP ribulose monophosphate  相似文献   

17.
Summary A 3.7 kb DNA fragment of yeast chromosome IV has been sequenced that contains the SFA gene which, when present on a multi-copy plasmid in Saccharomyces cerevisiae, confers hyper-resistance to formaldehyde. The open reading frame of SFA is 1158 by in size and encodes a polypeptide of 386 amino acids. The predicted protein shows strong homologies to several mammalian alcohol dehydrogenases and contains a sequence characteristic of binding sites for NAD. Overexpression of the SFA gene leads to enhanced consumption of formaldehyde, which is most probably the reason for the observed hyper-resistance phenotype. In sfa:LEU2 disruption mutants, sensitivity to formaldehyde is correlated with reduced degradation of the chemical. The SFA gene shares an 868 by divergent promoter with UGX2 a gene of yet unknown function. Promoter deletion studies with a SFA promoter-lacZ gene fusion construct revealed negative interference on expression of SFA by upstream sequences. The upstream region between positons – 145 and – 172 is totally or partially responsible for control of inducibility of SFA by chemicals such as formaldehyde (FA), ethanol and methyl methanesulphonate. The 41 kDa SFA-encoded protein was purified from a hyper-resistant transformant; it oxidizes long-chain alcohols and, in the presence of glutathione, is able to oxidize FA. SFA is predicted to code for a long-chain alcohol dehydrogenase (glutathione-dependent formaldehyde dehydrogenase) of the yeast S. cerevisiae.  相似文献   

18.
The phenology of citricola scale, Coccus pseudomagnoliarum (Kuwana), and its associated parasitoid complex were studied on citrus in the San Joaquin Valley of central California over the period April 1995–March 1997. A total of 10,237 parasitoid specimens of 10 species were collected. Two of these species, Marietta mexicana (Howard) and Encyrtus lecaniorum (Mayr), each recovered from individually isolated scales, represent new parasitoid records for citricola scale. A third species, Encarsia citrinus citrinus (Craw), may represent a new parasitoid record, but this requires further confirmation because a single (male) specimen was recovered from individually isolated scales. The three most dominant parasitoid species, Coccophagus lycimnia (Walker), Metaphycus helvolus (Compere), and Metaphycus luteolus (Timberlake), accounted for the majority (>97%) of the specimens recovered. In contrast to the situation on citrus in southern California, where citricola scale is under effective biological control and is very rarely seen, citricola scale on citrus in the San Joaquin Valley is reemerging as a major pest, especially in groves employing integrated pest management with minimal use of broad-spectrum insecticides. Possible reasons uncovered in this study for the lack of effective biological control of citricola scale in the San Joaquin Valley include: (i) reduced presence of Metaphycus spp. because of hyperparasitism by the heteronomous hyperparasitoid C. lycimnia; (ii) absence of alternate hosts for those species of Metaphycus present; and (iii) absence of hosts of suitable size for Metaphycus at critical times of the year. Recommendations for improving the level of biological control in the San Joaquin Valley are discussed.  相似文献   

19.
The influence of nitrogen limitation on the regulation of the methanol oxidizing enzymes alcohol oxidase, catalase, formaldehyde dehydrogenase and formate dehydrogenase in the two methylotrophic yeastsHansenula polymorpha andKloeckera sp. 2201 was studied in continuous culture. When shifted from carbon-limited growth conditions (with a mixture of glucose and methanol as carbon sources) to a nitrogen-limited environment both cultures were found to go through a transition phase where neither enhanced residual concentrations of the nitrogen source nor of one of the two carbon sources could be detected in the supernatant. As soon as nitrogen became a limiting substrate an immediate reorganisation of the cell composition was initiated: protein content of the cells dropped to approximately 40% of its initial value, glycogen was synthesized and the enzyme composition of the cells was changed. The peroxisomal enzymes alcohol oxidase and catalase in both organisms and the two dehydrogenases for formaldehyde and formate in cells ofKloeckera sp. 2201 were subject to degradation (catabolite inactivation). The measured rates of inactivation indicated that in cells ofH. polymorpha this process might be limited to peroxisomes, whereas inKloeckera sp. 2201 the degradation was found to affect peroxisomal as well as cytoplasmic enzymes. In contrast to methanol dissimilating enzymes the net rate of synthesis of hexokinase, glucose-6-phosphate dehydrogenase and 6-phosphogluconate dehydrogenase was not affected by this process but those enzymes were synthesized with increased rates.  相似文献   

20.
The earliest record in western North America of Trochammina hadai Uchio, a benthic foraminifer common in Japanese estuaries, is from sediment collected in Puget Sound in 1971. It was first found in San Francisco Bay in sediment samples taken in 1983, and since 1986 has been collected at 91% of the sampled sites in the Bay, constituting up to 93% of the foraminiferal assemblage at individual sites. The species is also present in recent sediment samples from 12 other sites along the west coast of North America. The evidence indicates that T. hadai is a recent introduction to San Francisco Bay, and is probably also not native to the other North American sites. Trochammina hadai was probably transported from Japan in ships' ballast tanks, in mud associated with anchors, or in sediments associated with oysters imported for mariculture. Its remarkable invasion of San Francisco Bay suggests the potential for massive, rapid invasions by other marine microorganisms.  相似文献   

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