Kinetic mechanism of the interaction of D-cycloserine with serine hydroxymethyltransferase

The kinetic mechanism for the interaction of D-cycloserine with serine hydroxymethyltransferase (EC2.1.2.1) from sheep liver was established by measuring changes in the activity, absorbance, and circular dichoism (CD) of the enzyme. The irreversible inhibition of the enzyme was characterized by three detectable steps: an initial rapid step followed by two successive steps with rate constants of 5.4 X s-l and 1.4 X lo4 s-l. The first step was distinguished by a rapid disappearance of the enzyme absorbance peak at 425 nm, a decrease in the enzyme activity to 25% of the uninhibited velocity, and a lowering of the CD intensity at 432 nm to about 65% of the original value. The second step of the interaction was accompanied by a complete loss of enzyme.

Human Torque teno virus: epidemiology, cell biology and immunology

Torque teno virus (TTV) was discovered in 1997 in the serum of a Japanese patient who had a post-transfusion hepatitis of unknown etiology. It is a small virus containing a circular single-stranded DNA genome which is unique among human viruses. Within a few years after its discovery, the TTVs were noted to form a large family of viruses with numerous genotypes. TTV is highly prevalent among the general population throughout the world, and persistent infections and co-infections with several genotypes occur frequently. However, the pathogenicity and the mechanism for the sustained occurrence of the virus in blood are at present unclear. To determine the prevalence of TTV in Finland, we set up PCR methods and examined the sera of asymptomatic subjects for the presence of TTV DNA and for genotype-6 DNA. TTV was found to be highly prevalent also in Finland; 85% of adults harbored TTV in their blood, and 4% were infected with genotype-6. In addition, TTV DNA was detected in a number of different tissues, with no tissue-type or symptom specificity. Most cell-biological events during TTV infections are at the moment unknown. Replicating TTV DNA has, however, been detected in liver and the hematopoietic compartment, and three mRNAs are known to be generated. To characterize TTV cell biology in more detail, we cloned in full length the genome of TTV genotype 6. We showed that in human kidney-derived cells TTV produces altogether six proteins with distinct subcellular localizations. TTV mRNA transcription was detected in all cell lines transfected with the full-length clone, and TTV DNA replicated in several of them, including those of erythroid, kidney, and hepatic origin. Furthermore, the viral DNA replication was shown to utilize the cellular DNA polymerases. Diagnoses of TTV infections have been based almost solely on PCR, whereas serological tests, measuring antibody responses, would give more information on many aspects of these infections. To investigate the TTV immunology in more detail, we produced all six TTV proteins for use as antigens in serological tests. We detected in human sera IgM and IgG antibodies to occur simultaneously with TTV DNA, and observed appearance of TTV DNA regardless of pre-existing antibodies, and disappearance of TTV DNA after antibody appearance. The genotype-6 nucleotide sequence remained stable for years within the infected subjects, suggesting that some mechanism other than mutations is used by this minute virus to evade our immune system and to establish chronic infections in immunocompetent subjects.

Electron- Spin-Resonance studies of phase transitions in double propionates

Detailed ESR investigations of Mn2+ substituting for Ca2+ in Ca2Sr(C2H5COO)6, (DSP) and Ca2Pb(C2H5COO)6, (DLP) and Ca2Ba(C2H5COO)6, (DBP), in single crystals and powders, over the temperature range from 300°C to -180°C have been carried out to study the successive phase transitions in these compounds. Spectra have been analyzed in terms of axial spin Hamiltonians and the temperature dependences of the parameters studied. Across the I-II transition, new physically and chemically inequivalent sites appear indicating the disappearance of the diad axes on which the propionate groups are located, bringing out the connection between the motional states of the propionate groups and the occurrence of ferroelectricity. The II-III transition also causes chemically inequivalent sites to develop, indicating that the transitions may not be isomorphous as believed previously. Similarities and dissimilarities of the ESR spectra of DLP, DSP and DBP are discussed in relation to the phase transitions.

On the Maximum Cloud Zone and the ITCZ over Indian, Longitudes during the Southwest Monsoon

An investigation is presented of the daily variation of the maximum cloud zone (MCZ) and the 7W mb trough in the Northern Hemisphere over the Indian longitudes 70–90°E during April–October for 1973–77. It is found that during June–September there are two favorable locations for a MCZ over these longitudes–on a majority of days the MCZ is present in the monsoon zone north of 15°N, and often a secondary MCZ occurs in the equatorial region (0–10°N). The monsoon MCZ gets established by northward movement of the MCZ occurring over the equatorial Indian ocean in April and May. The secondary MCZ appears intermittently, and is characterized by long spells of persistence only when the monsoon MCZ is absent. In each of the seasons studied, the MCZ temporarily disappeared from the mean summer monsoon location (15–28°N) about four weeks after it was established near the beginning of July. It is reestablished by the northward movement of the secondary MCZ, which becomes active during the absence of the monsoon MCZ, in a manner strikingly similar to that observed in the spring to summer transition. A break in monsoon conditions prevails just prior to the temporary disappearance of the monsoon MCZ. Thus we conclude that the monsoon MCZ cannot survive for longer than a month without reestablishment by the secondary MCZ. Possible underlying mechanisms are also discussed.

The decline of northern malaria and population dynamics of Plamodium vivax

Europe was declared malaria free in 1975. The disappearance of malaria has traditionally been attributed to numerous deliberate actions like vector control, the screening of houses, more efficient medication etc. Malaria, however, disappeared from many countries like Finland before any counter measures had even started. The aim of this thesis is to study the population ecology of P. vivax and its interaction with the human host and the vector. By finding the factors that attributed to the extinction of vivax malaria it might be possible to improve the modern strategy against P. vivax. The parasite was studied with data from Finland, which provides the longest time series (1749-2008) of malaria statistics in the world. The malaria vectors, Anopheles messeae and A. beklemishevi are still common species in the country. The eradication of vivax malaria is difficult because the parasite has a dormant stage that can cause a relapse long after a primary infection. It was now shown that P. vivax is able to detect the presence of a potential vector. A dormant stage is triggered even from a bite of an uninfected Anopheles mosquito. This optimizes the chances for the Plasmodium to reach a mosquito vector for sexual reproduction. The longevity of the dormant stage could be shown to be at least nine years. The parasite spends several years in its human host and the behaviour of the human carrier had a profound impact on the decline of the disease in Finland. Malaria spring epidemics could be explained by a previous warm summer. Neither annual nor summer mean temperature had any impact on the long term malaria trend. Malaria disappeared slowly from Finland without mosquito control. The sociological change from extended families to nuclear families led to decreased household size. The decreased household size correlated strongly with the decline of malaria. That led to an increased isolation of the subpopulations of P. vivax. Their habitat consisted of the bedrooms in which human carriers slept together with the overwintering vectors. The isolation of the parasite ultimately led to the extinction of vivax malaria. Metapopulation models adapted to local conditions should therefore be implemented as a tool for settlement planning and socio-economic development and become an integrated part of the fight against malaria.

ESR studies of phase transitions in double propionates

Detailed ESR investigations of Mn2+ substituting for Ca2+ in Ca2Sr(C2H5COO)6 (DSP), Ca2Pb(C2H5COO)6 (DLP) and Ca2Ba(C2H5COO)6 (DBP), in single crystals and powders, over the temperature range from 200°C to -180°C have been carried out to study the successive phase transitions in these compounds. (DSP: [Tetragonal] ← 8.5°C → [tetragonal, ferroelectric] [tetragonal] ← -169°C → [monoclinic, ferroelectric]; DLP : [tetragonal] ← 60°C → [tetragonal, ferroelectric] ← -71.5°C → [monoclinic, ferroelectric]; [Cubic] ← -6°C → [orthorhombic] ← -75°C → [?]). Spectra have been analysed in terms of axial spin Hamiltonians and the temperature dependences of the parameters studied. In DSP and DLP across the I ↔ II transition, new physically and chemically inequivalent sites appear indicating the disappearance of the diad axes on which the propionate groups are located, bringing out the connection between the motional states of the propionate groups and the occurence of ferroelectricity. The II ↔ III transition also causes chemically inequivalent sites to develop, indicating that the transitions may not be isomorphous as believed previously. In DBP, the -6°C transition leads to (i) a doubling of both physically and chemically inequivalent sites (ii) a small (150 G at -6°C to 170 G at -8°C), but abrupt change in the magnitude of the zero-field splitting tensor D, and (iii) displacements of the orientations of the D tensors. Results are interpreted in terms of alternate rotations of the oxygen octahedra, showing participation of the carboxyl oxygens in the transition. No drastic changes in the parameters occur across the -75°C transition consistent with its second order nature. Similarities and dissimilarities of the ESR spectra of the three compounds in relation to the phase transitions, are discussed.

Vanadate-stimulated NADH oxidation in plasma membrane

The rate of NADH oxidation with oxygen as the acceptor is very low in mouse liver plasma membrane and erythrocyte membrane. When vanadate is added, this rate is stimulated 10- to 20-fold. The absorption spectrum of vanadate does not change with the disappearance of NADH. The reaction is inhibited by superoxide dismutase, and there is no activity under an argon atmosphere. This indicates that oxygen is the electron acceptor and the reaction is mediated by superoxide. The vanadate stimulation is not limited to plasma membrane. Golgi apparatus and endoplasmic reticulum show similar increase in NADH oxidase activity when vanadate is added. The endomembranes have significant vanadate-stimulated activity with both NADH and NADPH. The vanadate-stimulated NADH oxidase in plasma membrane is inhibited by compounds, which inhibit NADH dehydrogenase activity: catechols, anthracycline drugs and manganese. This activity is stimulated by high phosphate and sulfate anion concentrations.

In vitro degradation of hepatotoxic indospicine in indigofera spicata by camel foregut fluid

Indospicine toxicosis was reported in sheep, goats and cattle fed on Indigofera, a leguminous plant rich in indospicine. Recent death report on dogs as a result of dietary ingestion of indospicine contaminated camel meat has raised concern about the distribution of this toxin in camels fed on Indigofera. This in vitro study aimed at measuring the degradability of indospicine in Indigofera spicata by camel-foregut fluid and attempted at explaining indospicine accumulation in meat tissue. In the first experiment, in vitro dry matter digestibility and indospicine disappearance were evaluated by using foregut fluid from 15 feral camels. Foregut fluid was collected post mortem from a nearby abattoir. In the second experiment, a composite foregut fluid obtained from three feral camels was used to examine the time-dependent degradation of indospicine. Results indicated that 99 of the dietary indospicine was degraded after 48 h of incubation. The time-dependent degradation study showed rapid degradation (11 µg/h) during the first 18 h of incubation, followed by a much slower rate (2 µg/h) between 18-48 h. Results demonstrated the ability of the camel microbiota to degrade indospicine and suggest the presence of a by-pass mechanism that enables the toxin to escape degradation and reaches the intestine.

On the Specificity of Carbohydrate-Lectin Recognition The Interaction of a Lectin from Ricinus communis Beans with Simple Saccharides and Concanavalin A

A galactose-specific protein (RC1) isolated from Ricinus communis beans was found to give a precipitin reaction with concanavalin A. Its carbohydrate content amounted to 8–9% of the total protein and was found to be rich in mannose. The interaction of RC1 with galactose and lactose was measured in 0.05 M phosphate buffer containing 0.2 M NaCl (pH 6.8) by the method of conventional equilibrium dialysis. From the analysis of the binding data according to Scatchard method the association constant (Ka) at 5°C was calculated as 3.8 mM−1 and 1.2 mM−1 for lactose and galactose, respectively. In both cases the number of binding sites per molecule of RC1 with molecular weight of 120000 was found to be 2. From the temperature-dependent Ka values for the binding of lactose, the values of –5.7 kcal/mol and –4.3 cal × mol−1× K−1 were calculated for ΔH and ΔS, respectively. The addition of concanavalin A to RC1 or vice versa led to the formation of the insoluble complex RC1· ConA4 containing one molecule of RC1 and one molecule of tetrameric concanavalin A (ConA4) which could be dissociated upon addition of concanavalin A-specific sugars. The complex formation results in a time-dependent appearance of turbidity in the time range from 10s to 10 min. From the measurement of the time-dependent appearance and disappearance of the turbidity the formation (kf) and dissociation (kd) rate constants were calculated as 3 mM−1× s−1 and 0.07 ks−1 respectively. The ratio kf/kd (43μM −1), that corresponds to the association constant of complex RC1· ConA4, is higher than that of mannoside · ConA4 and thereby suggests that protein-protein interaction contributes significantly in stabilising glycoprotein · lectin complexes. The relevance of this finding to the understanding of the chemical specificities that are involved in a model cell-lectin interaction is discussed.

Estrogen-induced synthesis of riboflavin-binding protein in immature chicks kinetics and hormonal specificity

The kinetics of estrogen-induced elevation in the plasma concentration of riboflavin-binding protein, a minor yolk constituent, was investigated in immature male chicks, using a specific and sensitive radioimmunoassay proceudre. Following a single injection of the hormone, the plasma riboflavin-binding protein content was enhanced several-fold at 6 h. reaching peak levels around 48 h and declining thereafter. A two-fold amplication of the response was evident on secondary stimulation with the hormone. A 4-h lag phase prior to onset of induction was noticed during both primary and secondary stimulat ions with the steroid hormone. The magnitude of the response was dependent on the hormonal dose whereas the initial lag phase and the time of peak riboflavin-binding protein accumulation were unaltered within the range of hormonal doses tested. The half-life of riboflavin-binding protein in the circulation was 10 h, as calculated from measurement of the rate of disappearance of exogenously administered 125I-labelled protein. Simultaneous administration of progestrone did bot affect the kinetics of riboflavin-binding protein production. On the other hand, the antiestrogens, cis- and trans-clomiphene citrates, given 30 min prior to estrogen and cycloheximide, effectively countered the hormone-induced riboflavin-binding protein elaboration. Both progesterone and the anti-esterogens per se were completely ineffective in substituting for estrogen in the inductive ptrocess.

Salaisen evankelistan salaliitto : Uusin keskustelu Klemens Aleksandrialaisen kirjeestä Theodorokselle

This thesis is an assessment of the hoax hypothesis, mainly propagated in Stephen C. Carlson's 2005 monograph "The Gospel Hoax: Morton Smith's Invention of Secret Mark", which suggests that professor Morton Smith (1915-1991) forged Clement of Alexandria's letter to Theodore. This letter Smith claimed to have discovered as an 18th century copy in the monastery of Mar Saba in 1958. The Introduction narrates the discovery story of Morton Smith and traces the manuscript's whereabouts up to its apparent disappearance in 1990 following with a brief history of scholarship of the MS and some methodological considerations. Chapters 2 and 3 deal with the arguments for the hoax (mainly by Stephen C. Carlson) and against it (mainly Scott G. Brown). Chapter 2 looks at the MS in its physical aspects, and chapter 3 assesses its subject matter. I conclude that some of the details fit reasonably well with the hoax hypothesis, but on the whole the arguments against it are more persuasive. Especially Carlson's use of QDE-analysis (Questioned Document Examination) has many problems. Comparing the handwriting of Clement's letter to Morton Smith's handwriting I conclude that there are some "repeated differences" between them suggesting that Smith is not the writer of the disputed letter. Clement's letter to Theodore derives most likely from antiquity though the exact details of its character are not discussed in length in this thesis. In Chapter 4 I take a special look at Stephen C. Carlson's arguments which propose that Morton Smith hid clues of his identity to the MS and the materials surrounding it. Comparing these alleged clues to known pseudoscientific works I conclude that Carlson utilizes here methods normally reserved for building a conspiracy theory; thus Carlson's hoax hypothesis has serious methodological flaws in respect to these hidden clues. I construct a model of these questionable methods titled "a boisterous pseudohistorical method" that contains three parts: 1) beginning with a question that from the beginning implicitly contains the answer, 2) considering everything will do as evidence for the conspiracy theory, and 3) abandoning probability and thinking literally that everything is connected. I propose that Stephen C. Carlson utilizes these pseudoscientific methods in his unearthing of Morton Smith's "clues". Chapter 5 looks briefly at the literary genre I title "textual puzzle -thriller". Because even biblical scholarship follows the signs of the times, I propose Carlson's hoax hypothesis has its literary equivalents in fiction in titles like Dan Brown's "Da Vinci Code" and in academic works in titles like John Dart's "Decoding Mark". All of these are interested in solving textual puzzles, even though the methodological choices are not acceptable for scholarship. Thus the hoax hypothesis as a whole is alternatively either unpersuasive or plain bad science.

A Raman study of multiferroic LuMnO3

Magnetic and dielectric measurements confirm the multiferroic nature of LuMnO3. Raman spectra of LuMnO3 have been recorded in the 77-800 K range covering both the antiferromagnetic transition at 90 K and the ferroelectric-paraelectric transition at 750 K. The changes in the phonon modes frequencies and band-widths indicate the presence of phonon-spin coupling in the antiferromagnetically ordered phase. The ferroelectric-paraelectric transition is accompanied by the broadening and disappearance of many of the phonon modes. Some of the phonon modes also show anomalies at the ferroelectric transition.

The Vexing Case of Igor Shafarevich, a Russian Political Thinker

The Russian mathematician, academician and former dissident Igor Shafarevich (b. 1923) is commonly mentioned in Western scholarly studies on perestroika and post-perestroika-era Russian politics as one of the most notable anti-Semites and extreme nationalists of the country. This notoriety owes to Shafarevich’s old samizdat article Russophobia, which was published in 1988. The scandal surrounding Russophobia came to a head when the president of The National Academy of Sciences in the United States asked Shafarevich, its honorary member, to resign. Nothing like this had ever happened in the academy’s history. The present dissertation discusses Shafarevich’s political activities, his texts and ideas as well as their reception. Particular attention is given to Russophobia, whose detailed examination proves very clearly that its reputation as an anti-Semitic text is groundless. The reasons for Russophobia’s hasty but fierce condemnation were many, but only one was that when the Soviet system began to tumble, it was commonly assumed that a vigorous rise in anti-Semitism and extreme nationalism in the Soviet Union/Russia would be just a matter of time. Many observers were highly sensitised to detecting its signs and symptoms. The dissertation also shows that most of those to write the first criticisms of Russophobia and to liken Shafarevich to the ideologues of Nazi Germany were the same people he had criticised in Russophobia for their deterministic view of history and irrational manner of connecting things for the purpose of fanning the flames of distrust between Russia’s Jews and Russians. In retrospect, it is fairly evident that Shafarevich actually managed to effectively “neutralise” the message of many of those obsessed with the Jews among his Russian contemporaries and contributed to the fact that anti-Jewish sentiments have been a great deal less popular in post-communist Russia than so many had feared and expected. The thesis also thoroughly discusses Shafarevich’s other texts and activities before Russophobia’s appearance and after it. In the 1970s, Shafarevich was one of the best-known dissidents in the Soviet Union. He worked together with academician Andrei Sakharov in a dissidents’ unofficial human rights committee and co-operated closely with Aleksandr Solzhenitsyn before Solzhenitsyn’s exile. Then, during the chaotic years of perestroika, Shafarevich defended the basic rights of ordinary citizens and warned that the hype concerning democracy could become counterproductive if the most palpable result of the reforms was the disappearance of citizens’ basic security and elementary social justice. One of the conclusions of the thesis is that even if the world around Shafarevich has changed considerably, his views have remained essentially the same since the late 1960s and early 1970s.

Oestrogen-induced synthesis of thiamin-binding protein in immature chicks. Kinetics of induction, hormonal specificity and modulation

A specific radioimmunoassay procedure was developed to monitor the plasma concentrations of thiamin-binding protein, a minor yolk constituent of the chicken egg. By using this sensitive assay, the kinetics of oestrogen-induced elaboration of this specific protein in immature chicks was investigated. After a single injection of the steroid hormone, with an initial lag period of 4–5h the thiamin-binding protein rapidly accumulated in the plasma, attaining peak concentrations around 75h and declining thereafter. A 4-fold amplification of the response was noticed during the secondary stimulation, and this increased to 9-fold during the tertiary stimulation with the steroid hormone. The magnitude of the response was dependent on the hormone dose, and the initial latent period and the duration of the ascending phase of induction were unchanged for the hormonal doses tested during both the primary and secondary stimulations. The circulatory half-life of the protein was 6h as calculated from the measurement of the rate of disappearance of the exogenously administered 125I-labelled protein. Simultaneous administration of progesterone, dihydrotestosterone or corticosterone did not alter the pattern of induction. On the other hand, hyperthyroidism markedly decreased the oestrogenic response, whereas propylthiouracil-induced hypothyroidism had the opposite effect. The anti-oestrogen E- and Z-clomiphene citrates, administered 30min before oestrogen, effectively blocked the hormonal induction. α-Amanitin and cycloheximide administered along with or shortly after the sex steroid severely curtailed the protein elaboration. A comparison of the kinetics of induction of thiamin- and riboflavin-binding proteins by oestrogen revealed that, beneath an apparent similarity, a clear-cut difference exists between the two vitamin-binding proteins, particularly with regard to hormonal dose-dependent sensitivity of induction and the half-life in circulation. The steroid-mediated elaboration of the two yolk proteins thus appears to be not strictly co-ordinated, despite several common regulatory features underlying their induction.

Underwater Sustainability of the ``Cassie'' State of Wetting

A rough hydrophobic surface when immersed in water can result in a ``Cassie'' state of wetting in which the water is in contact with both the solid surface and the entrapped air. The sustainability of the entrapped air on such surfaces is important for underwater applications such as reduction of flow resistance in microchannels and drag reduction of submerged bodies such as hydrofoils. We utilize an optical technique based oil total internal reflection of light at the water-air interface to quantify the spatial distribution of trapped air oil such a surface and its variation with immersion time. With this technique, we evaluate the sustainability of the Cassie state on hydrophobic surfaces with four different kinds of textures. The textures studied are regular arrays of pillars, ridges, and holes that were created in silicon by a wet etching technique, and also a texture of random craters that was obtained through electrodischarge machining of aluminum. These surfaces were rendered hydrophobic with a self-assembled layer Of fluorooctyl trichlorosilane. Depending on the texture, the size and shape of the trapped air pockets were found to vary. However, irrespective of the texture, both the size and the number of air pockets were found to decrease with time gradually and eventually disappear, suggesting that the sustainability of the ``Cassie'' state is finite for all the microstructures Studied. This is possibly due to diffusion of air from the trapped air pockets into the water. The time scale for disappearance of air pockets was found to depend on the kind of microstructure and the hydrostatic pressure at the water-air interface. For the surface with a regular array of pillars, the air pockets were found to be in the form of a thin layer perched on top of the pillars with a large lateral extent compared to the spacing between pillars. For other surfaces studied, the air pockets are smaller and are of the same order as the characteristic length scale of the texture. Measurements for the surface with holes indicate that the time for air-pocket disappearance reduces as the hydrostatic pressure is increased.