Statistical and frequency-amplitude characteristics of ultra-weak emissions of loach eggs and embryos under normal conditions and on their optic interactions. II. Changes in characteristics of ultra-weak emissions upon optic interaction of groups of embryos of different ages.

We compared the characteristics of ultraweak emissions from groups of loach embryos of different ages in the presence or absence of optic interaction. The percentage of zero values of emission gradually increased during the first hour of optic interaction. The number and height of rare big pulses estimated by the value of kurtosis increased in parallel. In addition, the correlation between the Fourier spectra of optically interacting samples decreased at a higher rate than in the absence of optical contact. Just after the 1-hour optic interaction was terminated, the number of high pulses decreased in a younger interacting group and increased in the older one and the farther away the partner groups were in developmental stages, the more pronounced these differences were. Measurements of the Fourier spectra after long-term (12-22-hour) optic interactions have shown that an "exchange" of autocorrelation characteristics of the spectra took place among the samples: the sums of autocorrelation coefficients were inverted in the vast majority of cases, often with an "overshoot" or, at least, were smoothed over with reference to the control samples. We conclude that the previously described effects of optic interactions between groups of loach embryos of different ages could be due to changes in the frequency spectra of their ultraweak emissions.     

Studies on Survival of Bacteria: Rhythmic Response of Microorganisms to Freeze-Drying Additives

Various materials were mixed with suspensions of Serratia marcescens and other organisms. Samples were removed and frozen at intervals after mixing; the number of cells that survived both freeze-drying and exposure to air varied rhythmically as a function of time between mixing and freezing. When assayed before or immediately after drying there were essentially no fluctuations. The response was evident only when these dried samples were exposed to air. In a typical experiment, the number of cells surviving in the sample frozen 30 sec after adding propyl gallate was at least 10 times that in samples frozen either 20 sec earlier or 20 sec later. Other "peaks" in survival were observed at approximately 125 and 450 sec, but the times at which the peaks were observed were not consistent from one experiment to the next. Although we have been unable to control or predict the time at which maxima in resistance occur, we have shown that the phenomenon does occur with Escherichia coli and Saccharomyces cerevisiae as well as with S. marcescens. Furthermore, a rhythmic response also was obtained after a change in pH or cell concentration. It appears that microorganisms respond physiologically and synchronously to changes in their environment, and some of these responses have survival value.     

On the Horizontal Distribution of the Profundal Bottom Fauna in a Limited Area of Central Lake Malaren,Sweden

The spatial distribution of bottom animals in a large, flat, and unexposed bottom area (about 8 km²) in central Lake Mälaren was investigated. As far as could be seen from preliminary tests the area was homogeneous in most respects. Working from the ice in all 42 tube samples and 6 Ekman samples were obtained from randomly distributed quadrats in a grid measuring 100 × 50 m². A 0.3 mm gauge net was used for sieving. The mud surface area covered with the tube samples was about 40 cm². Different methods of illustration were applied to demonstrate how many samples were actually required to show certain characteristics in the bottom fauna. For most purposes the accumulated mean value (cf. Kajak, 1963) was close enough to the stable mean value already after two or three tube samples for both chironomids and tubificids. In order to get a good idea of the five major constituents amongst tubificids (about 96% of all tubificids) two tube samples were found to be quite enough in this particular test. For the seven major constituents (about 99% of all tubificids) four tube samples were sufficient — or one single Ekman sample. The main constituents were well represented after two tube samples or one Ekman sample. Very little further information about the major constituents was obtained after the minimum number of samples mentioned above had been taken. A statistical study based on the same material was performed by a group of statisticians at the Institute of Statistics, University of Stockholm. Different statistical tests were applied to the material. Due to the relatively small number of observations nothing definitely could be said about the actual distribution of tubificids or chironomids in the area. The chironomids seemed to conform better to the Poisson distribution than to the Negative Binomial distribution, while the tubificids showed greater agreement with the Negative Binomial distribution.     

Statistical and Frequency-Amplitude Characteristics of Ultraweak Emissions of the Loach Eggs and Embryos under the Normal Conditions and upon Their Optic Interactions. 2. Changes in Characteristics of Ultraweak Emissions upon Optic Interaction of Groups of Embryos of Different Ages

We compared the characteristics of ultraweak emissions from groups of loach embryos of different ages in the presence or absence of optic interaction. The percentage of zero values of emission gradually increased during the first hour of optic interaction. The number and height of rare big pulses estimated by the value of kurtosis increased in parallel. In addition, the correlation between the Fourier spectra of optically interacting samples decreased at a higher rate than in the absence of optical contact. Just after the 1-hour optic interaction was terminated, the number of high pulses decreased in a younger interacting group and increased in the older one and the farther away the partner groups were in developmental stages, the more pronounced these differences were. Measurements of the Fourier spectra after long-term (12–22-hour) optic interactions have shown that an exchange of autocorrelation characteristics of the spectra took place among the samples: the sums of autocorrelation coefficients were inverted in the vast majority of cases, often with an overshoot or, at least, were smoothed over with reference to the control samples. We conclude that the previously described effects of optic interactions between groups of loach embryos of different ages could be due to changes in the frequency spectra of their ultraweak emissions.     

Factors influencing the cultivability of lake water bacteria

Counting bacteria in natural water samples by cultivation yields only low recovery efficiencies (ca. 1%), compared to total counts obtained after 4,6-diamidino-2-phenylindol (DAPI) staining. In order to optimize the cultivation of heterotrophic planktonic bacteria from Lake Constance (Germany), selected parameters of the medium composition were modified. The most important factor was the concentration of organic substrate (nutrient broth plus yeast extract), which significantly influenced the "most probable number" obtained in liquid growth medium. Reduced oxygen concentrations (3-12%) lowered the "most probable number". Addition of N-acyl homoserine lactones to the medium increased the cultivability slightly. Low substrate concentrations [0.03-0.06% (w/v)], an incubation atmosphere of 21% oxygen at 16 degrees C for 4 weeks were optimal and increased the cultivability ("most probable number" related to total bacterial counts) to an average cultivability of 18+/-11%, (n=8). The results indicate that cultivabilities of heterotrophic bacteria from lakewater samples can be significantly increased by modifying the cultivation methods.     

Does adipocyte hypercellularity in obesity exist?

Adipose tissue samples were biopsied from three subcutaneous sites in 80 obese and 27 non-obese patients. Additional samples were taken from intra-abdominal sites in 44 of the patients. There was a small increase in the calculated number of fat cells in the more obese patients, but there was no relation between fat-cell number and obesity of childhood onset. Omental fat cells were one-third the size of subcutaneous cells. Thus the calculated number of fat cells, usually based solely on subcutaneous samples, is an underestimate of the true number, and most obese patients can accommodate their fat without needing to recruit new cells. The diagnosis of "hyperplastic" obesity--that is, an excess number of fat cells--is unreliable and its relation to infantile obesity doubtful.     

南亚热带常绿阔叶林取样技术研究

 本文研究了样地取样技术和无样地取样技术在南亚热带常绿阔叶林的应用。认为中心点四分法无样地取样技术具有较高的精度和效率,完全适用于南亚热带常绿阔叶林。 应用中心点四分法无样地取样技术所获得的能反映群落真实特征的中心点数目,称为“最小点数”。评定最小点数,可应用种类数目与中心点数目的相关曲线,即“种一点数曲线”;以及优势种重要值与中心点数的相关曲线,即“重要值一点数曲线’。据研究南亚热带常绿阔叶林的最小点数为20—22点。 “重要值—面积曲线”与“重要值一点数曲线”是评定最小面积与最小点数的测度新方法,在某种意义上较“种一面积曲线”更为理想。     

Temporal Stability of the Salivary Microbiota in Oral Health

Objectives

Saliva is a biological fluid suitable for biomarker analysis, and differences in the salivary microbiota in oral health and disease have been reported. For such comparative analyses, time of sampling is critical since the bacterial composition may vary throughout the day, i.e., diurnal variation. The purpose of this study is to compare the salivary microbiome over time to determine the optimal time for sampling.

Design

Stimulated saliva samples were collected from 5 orally healthy individuals in 4 h intervals for 24 h, and collection was repeated 7 days later (number of samples per person, n = 12, total number of samples, n = 60). Salivary microbiota was analyzed using the Human Oral Microbe Identification using Next Generation Sequencing (HOMINGS), and statistical analysis was performed using the Kruskal-Wallis test with Benjamini-Hochberg’s correction for multiple comparisons, cluster analysis, principal component analysis and correspondence analysis.

Results

From a total of 60 saliva samples, 477 probe targets were collectively identified with a mean number of probes per sample of 207 (range: 153–307). Little or no variation in microbial profiles within subjects was observed over time.

Conclusions

Although there was considerable variation between subjects, microbial profiles within subjects were stable throughout a 24 hour period and after 1 week. Since there is little or no evidence of diurnal variation of the salivary microbiome, time of sampling of saliva is not critical for perturbation or other microbial studies.     

Rapid screening method for determination of Ecstasy and amphetamines in urine samples using gas chromatography-chemical ionisation mass spectrometry

The need for analytical screening tests more reliable and valid to detect amphetamine and related "designer drugs" in biological samples is becoming critical, due to the increasing diffusion of these drugs on the European illegal market. The most common screening procedures based on immunoassays suffer a number of limitations, including low sensitivity, lack of specificity and limited number of detectable substances. This paper describes a screening method based on gas-chromatography-mass-spectrometry (GC/MS) using positive chemical ionisation (PCI) detection. Methanol was used as reactant gas in the ionisation chamber. Molecular ions of different compounds were monitored, allowing a sensitivity of 5-10 ng/ml with high selectivity. The sensitivity of the method gives positive results in samples taken 48-72 h after intake of one dose of 50-100 mg. The method is simple and rapid. Sample preparation was limited to one liquid-liquid extraction, without any hydrolysis and derivatisation. Hydrolysis is critical to identify metabolites excreted as conjugates. Blank urine samples spiked with known amounts of amphetamine (AM), methylamphetamine (MA), methylenedioxyamphetamine (MDA), methylenedioxymethylamphetamine (MDMA), methylenedioxyethylamphetamine (MDEA) and methylenedioxyphenyl-N-methyl-2-butanamine (MBDB) were analysed. The method was successfully tested on real samples of urine from people, whose use of amphetamine was suspected, and results were compared with results obtained with immunoassays.     

Use of drift substrates to characterize marine fungal communities from the west coast of Portugal

This survey reports the occurrence, diversity and similarity of marine fungi associated with five categories of drift substrates (Arundo donax, Phragmites australis, Spartina maritima, "other stems" and driftwood) collected on four sandy beaches of the western coast of Portugal. "Other stems" and driftwood are composite samples with a variety of identified and unidentified pieces of non-woody and woody substrates respectively. Fifty-six taxa were identified, including 38 Ascomycota and 18 anamorphic fungi. Twenty-six taxa were generalists; however several cases of "substrate recurrence" were identified. The very frequent fungi differed among the categories of studied substrates, with the exception of Corollospora maritima, very frequent on four categories. Except for S. maritima, P. australis and driftwood, cases of multiple fungal colonization were rare. S. maritima was the single substrate with five different marine fungi on one sample, as well as with the highest number of very frequent fungi, highest percentage of colonization and average number of fungi per sample. Driftwood presented the highest value of fungal richness (37 taxa) and A. donax the lowest (22 taxa). ANOSIM analysis of similarity showed that all substrates supported different fungal communities with the exception of the pair P. australis/"other stems". The effect of sample size on estimated fungal richness was tested, and the results let us conclude that, although most of the sporadic fungi (<1% occurrence) will be detected only in a very large number of samples, 60 samples of A. donax and "other stems" and 70 samples of all the other substrates may suffice to assess their respective representative marine mycota.     

Identification and comparative analysis of microRNAs in <Emphasis Type="Italic">Pinus massoniana</Emphasis> infected by <Emphasis Type="Italic">Bursaphelenchus xylophilus</Emphasis>

Pine wilt disease seriously endangers pine tree growth and can lead to wilting and death. In this study, Masson pine (Pinus massoniana) was used as the donor plant to study miRNA expression by high-throughput sequencing in needle leaves of trees during the first 3 days after infection by Bursaphelenchus xylophilus. The results showed that the number of differentially expressed miRNAs first increased and then decreased in samples within 3 days of infection, with the highest expression in samples collected on day 2. Analysis of miRNA target genes in needles on different days after infection revealed enrichment in two pathways: plant hormone signal transduction and RNA transport. Further comparison of the samples indicated differential expression of 10 miRNAs on different days after infection, and the corresponding target genes, such as those participating in plant hormone signal transduction, were also significantly enriched. In addition, the concentrations of zeatin and indole acetic acid in needle tissues were significantly lower after infection than in the control sample. The above results indicated that B. xylophilus infection influenced the expression of miRNAs related to plant hormone signal transduction in P. massoniana, decreasing plant hormone synthesis and ultimately affecting the growth of Masson pine.     

The use of infants in drug research when there is no direct benefit to them: a survey of Canadian health care professionals serving on ethics committees.

Canadian health care professionals and lawyers serving on ethics committees were questioned about their views on pharmacokinetic research in newborn infants who are not likely to benefit directly from the results. Of the 50 respondents 13 felt that blood samples should be taken only for therapeutic reasons; 10 of the 13 argued that additional blood samples should not be taken, because there is no direct benefit to the infant; and 8 felt that proxy consent cannot be given for invasive nontherapeutic research. Four of the five participating lawyers would not permit additional blood samples to be taken. Of the 37 respondents who would permit additional blood samples to be taken, 27 felt that the number of samples taken should depend on the researcher''s justification for that number of samples; only 7 of the respondents had a clear idea of what the "upper limit" of the number of blood samples should be.     

Notes on the question of the specificity of reduction sites in the bacterial cell

The number of oxidoreduction sites and the total reducing activityof Proteus vulgaris cells was studied by means of Nitro blue tetrazolium after incubation with two substrates. The second substrate was added after ten minutes’ incubation with the first and the given quantities were determined in both samples. The following systems were used: glucose-formate, formate-glucose, pyruvate-lactate, lactate-pyruvate. In the controls, the initial substrate was re-added. In every case, the addition of another substrate was followed by a significant increase in the number of reduction sites and in the reducing activity of the cells as compared with the control. There are two possible explanations of this; either specific sites exist, for individual substrates, or the bacterial cell leaves a given number of “free places” for further substrates.     

Multiple amounts of DNA related to the size of chloroplasts

Summary Plants containing genetically small or large plastids appear in some euploid and trisomic types of Beta vulgaris.Small tissue samples of the first 6–10 leaves of nearly 30 euploid plants from 7 different generations were incubated in a solution containing ³H-thymidine in dark/light cycles, for up to 72 hr. For semiquantitative autoradiography the chloroplasts were then prepared on the slides with various kinds of isolation media, and fixed with OsO₄, glutaraldehyde, formaldehyde, or ethanol. The specifity of incorporation was tested by observing the extractability of label after differential treatment with acid or nucleases.Chloroplasts in leaves 2–11 cm long preferentially incorporate ³H-thymidine. The silver grains over plastids appear to be in clusters (centres). A relationship between the number of grains and also between the number of centres on the one hand, and the chloroplast size on the other could be found.It is concluded that chloroplasts occur in various degrees of polyenergide organization, as has been described, for example, for blue-green algae. Regarding the presence and degree of polyploidy—the other form of genetic polyvalency —the experiments provided no information. A remarkable variation in chloroplast size (and number of labelled centres) was observed, not only between different plants or between different leaves of a plants, but also within small tissue samples.     

Diversity of the microscopic fungi in the littoral sands of the White Sea

For the first time, the study of the mycobiota of littoral sands of the cold-water White Sea was conducted. Twenty-four samples were collected in July–August 2015 in the Onega Bay, the Kandalaksha Bay, and on the Terskii coast. Culture methods were used to detect the number of colony forming units (CFU) and the fungi diversity. The number of CFUs is small; it ranges from 0 to 57 per 1 cm³. A total of 54 morphotypes were detected, of which 22 were identified to the species, seven to the genus level, and 25 were not identified, because they did not form sporulation. Only two species, Cladosorium sphaerospermum and Paradendryphiella salina, were characterized by the relative abundance of more than 10% and the frequency of occurrence of more than 50%. Another five species were found in more than one sample: Penicillium chrysogenum, Cadophora fastigiata, Acremonium fuci, anamorph of Pseudogymnoascus pannorum, and teleomorph of Pseudeurotium hygrophilum. All other morphotypes were found in a small number in single samples. In general, darkcolored forms predominated in the mycobiota both in diversity and in number.     

The Early Endocrine Stress Response in Experimental Subarachnoid Hemorrhage

Introduction

In patients with severe illness, such as aneurysmal subarachnoid hemorrhage (SAH), a physiologic stress response is triggered. This includes activation of the hypothalamic-pituitary-adrenal (HPA) axis and the sympathetic nervous system. The aim of this study was to investigate the very early responses of these systems.

Methods

A porcine animal model of aneurysmal SAH was used. In this model, blood is injected slowly to the basal cisterns above the anterior skull base until the cerebral perfusion pressure is 0 mm Hg. Sampling was done from blood and urine at -10, +15, +75 and +135 minutes from time of induction of SAH. Analyses of adrenocorticotropic hormone (ACTH), cortisol, aldosterone, catecholamines and chromogranin-A were performed.

Results

Plasma ACTH, serum cortisol and plasma aldosterone increased in the samples following induction of SAH, and started to decline after 75 minutes. Urine cortisol also increased after SAH. Urine catecholamines and their metabolites were found to increase after SAH. Many samples were however below detection level, not allowing for statistical analysis. Plasma chromogranin-A peaked at 15 minutes after SAH, and thereafter decreased.

Conclusions

The endocrine stress response after aneurysmal SAH was found to start within 15 minutes in the HPA axis with early peak values of ACTH, cortisol and aldosterone. The fact that the concentrations of the HPA axis hormones decreased 135 minutes after SAH may suggest that a similar pattern exists in SAH patients, thus making it difficult to catch these early peak values. There were also indications of early activation of the sympathetic nervous system, but the small number of valid samples made interpretation difficult.     

Antagonistic effect of enterocin CCM 4231 from Enterococcus faecium on "bryndza", a traditional Slovak dairy product from sheep milk

"Bryndza" is a traditional Slovak dairy product (type of soft cheese) made from sheep cheese which was ripened for 14 days. Because its manufacture, transporting and/or storing represent conditions which facilitate contamination, the effect of enterocin CCM4231 in "bryndza" was investigated with the aim to reduce the contaminant agents. "Bryndza" was divided into equal portions (50 g). The experimental sample (ES) as well as the control sample one (C1) were inoculated with Listeria innocua Li1 strain. The other control samples C2 and C3 were without Li1 strain. C3 control was selected as a reference control. ES and C2 portions were treated with purified enterocin CCM4231 in a concentration of 6400 AU/ml. Before the experimental inoculation, "bryndza" was checked for the presence of contaminant agents. The experiment lasted 1 week and the samples were stored in the refrigerator at 4 degrees C. Sampling was performed on day 1, on day 4 and on day 7. The control samples C2 and C3 were checked only on day 1 and then after 1 week. The following contaminant agents were detected in "bryndza" before its experimental inoculation with L. innocua Li1 strain: Escherichia coli in the amount 10(3) cfu/ml/g, Staphylococcus aureus (10(2) cfu/ml/g) and enterococci (10(4) cfu/ml/g). In the control sample C2, the number of E. coli was reduced to 10(2) cfu/ml/g. Enterococci and staphylococci were totally eliminated there. Concerning C3 control, natural decrease of bacteria was found and/or their unchanged counts. The value of pH (5) was stable during the whole experiment. In the experimental sample inoculated with Li1 strain, its counts were decreased immediately after enterocin CCM4231 addition approximately by one order of magnitude. This inhibitory effect was also detectable on day 4 by the difference of one order of magnitude between ES and C1. On day 7, 10(3) cfu/ml/g of Li1 strain were detected in both samples (ES, C1). The difference by one order of magnitude indicated, an inhibitory effect of enterocin CCM4231 in "bryndza". However, bacteriocin activity was not determined by laboratory analyses.     

Studies on rats with islet beta cell tumors induced by nicotinamide and streptozotocin.

Islet beta cell adenomata were induced in rats by combined treatment with nicotinamide and streptozotocin. Three weeks after treatment marked alterations in glucose tolerance were noted in animals which later exhibited large beta cell tumors. Eight months after treatment, the rats known to have beta cell tumors on the basis of marked hypoglycemia and later confirmed by autopsy showed variable response to a glucose load. Some tumor-bearing rats showed fast response to glucose load, their blood sugar levels were elevated moderately and returned to normal or below normal levels rapidly; these animals are described as having "fast-acting tumors". Rats with "slow-acting tumors" responded sluggishly to a glucose load; their blood glucose pattern was similar to that of subdiabetic animals. Animals with beta cell tumors exhibited elevated serum insulin levels 30 min after glucose administration. Insulin biosynthesis by beta cell adenomata was demonstrated by in vitro incorporation of [14C]leucine into proinsulin and insulin. In the small number of tumor samples studied, a stimulatory effect of glucose on insulin biosynthesis was observed.     

Accessibility of sulfhydryl groups to 5,5'-dithiobis-2-nitrobenzoic acid and acid-base properties of bovine and walleye pollock rhodopsin preparations]

Both the number of exposed SH-groups and the rate of reaction with 5,5'dithiobis-2-nitrobenzoic acid (DTNB) in walleye pollock and bovine rhodopsin depend on a degree of native structure of the preparation to be investigated. The preparations studied can be arranged in the order of increase of these parameters as follows: ROS less than rhodopsin extracted by digitonin less than triton X-100 less than cetyltrimethylammonium bromide (CTAB) less than sodium dodecylsulphate (SDS). After illumination of ROS and digitonin, triton X-100 and CTAB-solubilized rhodopsin, and increase was observed in the number of modified SH-groups. Dark and bleached samples of walleye pollock rhodopsin exhibited a faster rate reaction and a more number of modified SH-groups as compared to bovine preparation. The differences between bovine and walleye pollock preparation disappeared after complete opsin unfolding as a result ROS solubilization in SDS. Six SH-groups per molecule of rhodopsin were modified in both preparation under these conditions. No differences in the number of cysteine residues (10--11), disulfide groups (2), acid (35--40) and base (25--30) titratable groups per rhodopsin molecule were found between bovine and walleye pollock ROS membranes. The isoelectric point of both rhodopsin preparations was within the pH range 5.2--5.6. After proteolysis of ROS with papain, a fragment with molecular weight 24500 +/- 1000 was detected, which contained the same number of SH-groups and cysteine residues as in the case of intact rhodopsin. The results obtained suggest that, in spite of a similar primary structure, the walleye pollock visual pigment has more "loose" and "fluid" space packing in the ROS membrane than the bovine pigment.     

Stress-induced cortisol is associated with generation of non-negative interpretations during cognitive reappraisal

Background

Enhanced processing of emotional stimuli after stress exposure is reported to be associated with stress-induced cortisol. Because enhanced emotional information processing could make cognitive emotion regulation more difficult, it was hypothesized that stress-induced cortisol would be associated with non-negative interpretation generation associated with the cognitive reappraisal processes.

Methods

A total of 36 participants (Mean age?=?21.3 years, SD?=?1.8) watched video clips of depression-related stressful situations before and after the administration of a stress induction task. They were then asked to generate as many non-negative interpretations as possible to reduce the depressive mood. Saliva samples were obtained before and after the stress induction task to measure change in the cortisol level.

Results

Participants were allocated post-hoc to either a responder (n?=?19) or non-responder group (n?=?17) based on the cortisol response to the stress induction task. The number of non-negative interpretations generated following the stress induction task was reduced only in the cortisol responders. The number of post-stress non-negative interpretations was fewer in the responder group when compared by sex, baseline cortisol level, and the number of pre-stress non-negative interpretations, statistically controlled.

Conclusions

Although baseline cortisol and sex may have impacted the results, the results suggest that stress-induced cortisol is associated with difficulty in non-negative interpretation generation during the cognitive reappraisal process.