Single-cell analysis of the physiology of mechanosensation in bacteria

Escherichia coli is one of the best studied living organisms and a model system for many biophysical investigations. Despite countless discoveries of the details of its physiology, we still lack a holistic understanding of how these bacteria react to changes in their environment. One of the most important examples is their response to osmotic shock. One of the mechanistic elements protecting cell integrity upon exposure to sudden changes of osmolarity is the presence of mechanosensitive channels in the cell membrane. These channels are believed to act as tension release valves protecting the inner membrane from rupturing. This thesis presents an experimental study of various aspects of mechanosensation in bacteria. We examine cell survival after osmotic shock and how the number of MscL (Mechanosensitive channel of Large conductance) channels expressed in a cell influences its physiology. We developed an assay that allows real-time monitoring of the rate of the osmotic challenge and direct observation of cell morphology during and after the exposure to osmolarity change. The work described in this thesis introduces tools that can be used to quantitatively determine at the single-cell level the number of expressed proteins (in this case MscL channels) as a function of, e.g., growth conditions. The improvement in our quantitative description of mechanosensation in bacteria allows us to address many, so far unsolved, problems, like the minimal number of channels needed for survival, and can begin to paint a clearer picture of why there are so many distinct types of mechanosensitive channels.

A dynamic CGE modelling approach for analyzing trade-offs in climate change policy options: the case of Green Climate Fund

38 p.

Comparative investigations on the respiratory physiology of euryhaline gammarids with special reference to salinity adaptation. [Translation from: Helgolaender Wissenschaftliche Meeresuntersuchungen 23(4) 485-534, 1972.]

Members of the family Gammaridae are very closely interrelated. There arises the question as to how far they also differ amongst themselves through physiological characteristics. Comparative respiratory and physiological experiments were made on the five euryhaline species Gammarus locusta, G. oceanicus, G. salinus, G. zaddachi and G. duebeni. The respiratory measurements carried out within the framework of this experiment were occupied with the relationships between oxygen consumption and body size depending on salinity. They also had the object of determing the variations in metabolic intensity after an abrupt change in the salt content of the external medium, and to establish the period of time for the process of adaptation. As the experiments were carried out polarographically in a testing plant with continuous flow-through, and the method which was applied permitted continuous recording over prolonged intervals, there could also be carried out comparisons between metabolism at rest and under activity, and the alterations of oxygen consumption during the process of moulting could be measured.

Physiology of nitrate reduction in Pseudomonas aeruginosa. [Translation from: Z.allg.Mikrobiol. 13, 83-94, 1973.]

Pseudomonas aeruginosa has for some time been known as a denitrifier. Pseudomonas aeruginosa was chosen for further studies, because P. aeruginosa occurs abundantly in Plusssee and moreover there are contradictory assertions on the gas products of denitrification by this bacteria. In experimental research the pattern of growth and gas production of Pseudomonas aeruginosa on nutrient broth was studied.

The effects of environmental stress on the physiology of growth in rainbow trout, Salmo gairderi Richardson

There is little doubt that both mammalian and teleost growth hormones can accelerate growth and increase food conversion efficiency in all commonly-reared species of salmonid fish. In those vertebrates that have been closely studied (predominantly mammals), the pituitary hormone somatotropin (GH or growth hormone) is a prime determinant of somatic growth. The hormone stimulates protein biosynthesis and tissue growth, enhances lipid utilization and lipid release from the adipose tissues (a protein-sparing effect) and suppresses the peripheral utilization of glucose. The present study is a prerequisite for future work on growth hormone physiology in salmonids and should contribute to our understanding of the mechanisms of growth suppression in stressed fish. Plasma growth hormone (GH) levels were measured in rainbow trout using a radioimmunoassay developed against chinook salmon growth hormone.

Feeding trials of green seaweed Ulva fasciata

For the nutritional evaluation of green seaweed Ulva fasciata, a feeding trial was performed in albino rats. The results indicated that the 20% replacement of seaweed U. fasciata instead of carrot or lettuce in rad diet causes no harmful effects, as evident by a non-significant change in blood constituents and serum enzyme levels. The weight gain observed in rats with U. fasciata diet was same as that of control diet. The true digestibility ratio of U. fasciata was 80.20% with carrot and 83.4% with lettuce. The data suggest that the green seaweed U. fasciata could be used as an alternative dietary component in animal fodder.

Culture of green algae Chlorella ellipsoidea in inexpensive media

The culture of the of green alga Chlorella ellipsoidea was conducted under natural conditions at the same place simultaneously in five different media, viz., medium-I (inorganic medium), medium-II (powdered whole-pulse medium), medium-III (medium of pulse bran), medium-IV (mixed medium = 50% inorganic medium + 50% whole-pulse powder medium), medium-V (mixed medium = 50% inorganic medium + 50% pulse bran medium). The culture was done in 500 ml conical flask. Growth rates of C. ellipsoidea in five different media were different and reached maximum cell densities of 0.63 x 10^6 cells per ml in 8 days in medium-I, 4.02 x 10^6 cells per ml in 10 days in medium-II, 3.62 x 10^6 cells per ml in 9 days in medium-III, 4.38 x 10^6 cells per ml in 11 days in medium-IV and 4.36 x 10^6 cells per ml in 11 days in medium-V. The range of air temperature was 20 to 33°C and that of culture media was 24 to 32°C and light intensity was 2000 to 7000 lux during the culture period. The inexpensive culture media were found to be significantly useful for algal culture.

Effects of hydrocarbons on reproductive physiology of the pearl oyster, Pinctada fucata

There are a lot of evidence that show hvdrocarbones cause some defect in reproduction and growth of bivalves. Bivalves are filter-feeder, thus accumulate more hydrocarbones in their tissue. In this study adult pearl producing oysters (Pinctada fucata) are used for all experimens. Samples of oysters, water and sediment from four natural beds; Nakhiloo (clean), Hendurabi (semipolluted), Lavan 1 (semipolluted) and Lavan 2 (polluted) were gatherd for 13 succesive months. Temperature, salinity, pH, oxygen and turbidity were recorded in each sampling. Oysters were kept in laboratory for adapation and then their length (DVM) were measured. Hemolymph samples were collected by insuline syring. Sediments and soft tissues of oysters were dissolved in carbon tetrachloride and when heated to extract oil hydrocarbones. UV, GC and IR were used to assay oil hydrocarbones. Accumulation of hydrocabones in soft tissue were as follows : Kakhilooof Lavan 1 and Kakhiloo station to the coast pollution was less than Hendurabi. Results showed that there is not meaningful difference (P<0.05) between sexual hormones in males and femals. Concerning progesterone cycle, there are two peak of spawning in oysters, major one in late spring and minor one in mid fall. Little elevation of progesterone start gamete and a lot of elevation release gametes_ Esteradiols gradually increase during gametogenesis and reach to maximum level during vitellogenesis. Testosterone have a synergestic role which esteradiol during vitellogenesis and also is effective in male sexuality. Multi regression test showed that there isn't meaning relationship (P<0.05) between hydrocarbones pollution and esteradiols. However in Lavan 2 due to hormone concentration of hydrocarbones had some effect in sexual hormonal cycles. Turbidity was the most effective factor for releasing of progesterone. Progesterone was a stimulating factor for releasing estradiol also release of testosterone was corrolated with oxygen and depth of beds. One way analysis of variance showed that there is not significant relasionship (P<0.05) between different factory in stations.

Reproductive physiology of ribbon fish (Trichiurus lepturus)

The ribbon fishes ‘of the family Trichiuridac are represented as one of the most important food resources in Indian ocean. High density of the dominant species of ribbon fish (Trichiurus lepturus) in Oman sea and the 'Tillable catch in last yeas (more than 7000 tones per year) makes a trust area for studing their population biolog and stock assessment. As our knowledge on reproductive biology of this species has an important role on their fisheries management, as well as conservation of this stock from decline or over fishing, this research was held to determine some aspects of reproductive physiology of ribbon fish and the effects of environmental factors in gonadal cycle. The goals of the present thesis is to determine some aspects of reproductive physiology such as gonadosomatic index (GSI) , hepatosomatic index (HSI), condition factor (Ko, fecundity, sex ratio, size at first maturity, size at maturity (LM5O) and their relative hormonal & biochemical fluctuations. In this regards annual variation of sex hormones ic. estradiol 17-B, progestron, cortisol, testostrone and gonadotropins FSH (GTH-I) , LH (GTH-ll)I were measured ; gonadal histological studies were done by light & electron micrography. The research was carried out from April 1995 to January 19% in Ras Nleidani in the north part of Oman sea, and the environmental factors such as temperature, salinity, oxygen, rainfall and pH were measured. The effects of these parameters on reproductive cycle and hormonal fluctuationswere discussed by using correlation and principle component analysis (PCA). Female Ribbon fish reproductive strategy shows the same paterns of nonguarder marine teleosts. T. lepturus has more than one spawning season (existance of egges in different size in each month) and therfore it must have asynchronous ovaries and belong to continious spawners. GSI and HSI are good evidences for this type of reproductive patern. The testis of the lobular type , which is typical of most teleosts , is composed of numerous lobules which are separated from each other by a thin layer of fibrous connective tissue. GSI fluctuations revealed prolong- spawning time in males. There is significant increase in 17-13 estradiol. progestrone , cortisol and gonadotropins with maturity and prespawning period of female T lepturus. Plasma concentration of E2 and GTH II incresaed along with water temperature increasing (3300).. Spawning was observed from Nov. 1995 to Apr. 1996 in this species. Progestrone increased significantly with increasing rainfall in this season (P<0.01). Plasma cortisol levels increased with maturation and vitelpgenesis and also with the peak of spawning. From lenght-weight frequency and size distribution in each age groups and also minimum size at first maturity (52a cm) it would he concluded that T. lepturus must be matured at 2 years of age. Serum cholestrol and triglicerides significantly increased when maturation occured in this species. The relationship between alkaline phosphatase activity and hormonal fluctuations with maturity and vitelogenesis were discussed. Proximate compostion (muscle) shows significant variation with spawning period and maturity. Absolute individual fecundity (17420-159150) increased with body length and weight. Ultrastructural observations show dramatic variation in cell membrane (0ocyte membrane), yolk vesicles and, nucleolus dispersal in relation to maturity stages. fluctuations of gonadal hormones were discused in relation with vitelogenesis. Testosterone increased in males from Nov: to Mar. due to environmental impacts and spawning time. Sex ratio in different depth (10-40 m ,80-110 m) shows significnt differences in this ratio for two depths. In 10-40 m depth female shows dominant abundance to male in each months that may be due to their reproductive migration behaviour. The effects of temperature photoperiod and rainfall to maturity and spawning were discussed. According to -pawning period of T. leptunts in our sampling area it could be suggested that ribbon fish fi,theries must be restricted in the peak of spawning seasons (Feb. to Mar.) and in the spawning grounds (under 40 m depths). Other suggestions for population conservation have been mentioned.