Anticoagulant, Fibrinolytic, and Antithrombotic Effects of the Heparin–Insulin Complex

Formation of heparin–insulin complex at the 1 : 10 molar ratio of the components has been demonstrated by spectral methods. The derived complex had anticoagulant, antithrombotic, and fibrinolytic properties of nonenzymatic nature in vitro. Intravenous injection of the complex in the animals (rats) increased anticoagulant and fibrinolytic background and at the same time decreased the plasma coagulation factors fibrinogen and factor XIIIa in the bloodstream. We propose the heparin–insulin complex as a promising antithrombotic drug.     

Accomplishments of the Trustees and laboratory staff of the Biological Stain Commission, 2002–2013

During the 12 years from 2002 to 2013, the Trustees and laboratory personnel of the Biological Stain Commission (BSC) can claim many accomplishments. These accomplishments are itemized under 11 categories: continuous publication of the official journal, Biotechnic & Histochemistry; production of four special issues of Biotechnic & Histochemistry devoted to specific dyes or stains; standardization of staining and dye purity; mechanisms of staining and prediction of dye behavior; publication of books or book chapters; effects of fixation and processing on staining; cancer research; immunohistochemistry; BSC Laboratory activities; miscellaneous publications; and administrative accomplishments.     

The Evolution of the idea of homeostasis: Determinism,stochastics, and chaos–self-organization

Homeostasis is a basic concept in investigating all living systems. Homeostasis and life are synonyms to a certain extent. The concept of homeostasis has been studied and developed for more than 150 years; however, only the 21st century brought us close to understanding homeostasis as a state of a biosystem and a continuous process. The chaos and self-organization theory (CSOT) proves that the conventional views based on determinism (functional analysis) or stochastics (when stochastic uncertainty or certainty occurs) cannot describe homeostasis as defined by W.B. Cannon and his followers. The new CSOT shows a clear boundary between determinism, stochastics (and deterministic chaos), and third-type systems, which have five special properties (principles of self-organization) and can be described in terms of quasiattractors. Kinematics is understood as the motion of quasi-attractors in the phase spaces of states. Complexity is introduced as the rate of evolution of complex biological systems, differing from the definition by Prigogine–Glansdorff. At the same time, concepts of uncertainty of the first and second types and an analog of the Heisenberg calculus are introduced for complexity. According to these concepts, complex biosystems are transferred from the area of traditional science to a new area of the theory of chaos and self-organization.     

The composition,structure, and dynamics of the nekton of the upper epipelagic layer in the Aleutian and Commander basins,western Bering Sea,in the fall seasons of 2002–2013

The data on the composition and abundance of nekton species and their interannual variations within the upper epipelagic layer (0–50 m) in the Aleutian and Commander basins in the western Bering Sea that are considered in the article were collected during complex surveys that were conducted by the Pacific Research Fisheries Center in September and October of 2002–2013. The core species structures of the nekton communities in these two areas were similar: the chum salmon Oncorhynchus keta and the boreopacific gonate squid Boreoteuthis borealis were the most abundant species. Simpson’s dominance index varied synchronously in both areas, with higher values in the Aleutian Basin, until 2009. In the subsequent years, the values of the index for the studied areas became equal and varied asynchronously. An analysis of abundance showed that two types of species structure most often prevailed (with the dominance of either chum salmon or squid); in some years the species structure differed from that in other years. Species of the low-boreal and low-boreal-subtropical complexes were more abundant in the Commander Basin, reaching particularly high proportions in 2006, 2008, and 2012. Certain species showed similarity in the year-to-year dynamics of their abundance; however, these coincidences were frequently accidental. After the climate regime shift in 2006–2007, the total biomass of nekton in the Aleutian Basin decreased from 3241 to 1736 kg/km² (46%); in the Komandorskiye Basin it decreased from 2459 to 1976 kg/km² (20%).     

Biologists and the promotion of birth control research, 1918–1938

Conclusions In spite of these efforts in the 1920s and 1930s to initiate ongoing research on contraception, the subject of birth control remained a problem of concern primarily to the social activist rather than to the research scientist or practicing physician.⁸⁰ In the 1930s, as has been shown, American scientists turned to the study of other aspects of reproductive physiology, while American physicians, anxious to eliminate the moral and medical dangers of contraception, only reluctantly accepted birth control as falling within their professional domain.As a result, the problem of cheap, effective, and safe contraception was not solved by these earliest attempts. Consideration of the subject was initiated afresh by private philanthropy after World War II, sparked by a new wave of interest in population studies.⁸¹ Summarizing such efforts to support research in the reproductive sciences, a recent Ford Foundation study has noted: To initiate and sustain serious research in the reproductive sciences has required for more than half a century concerted effort by interested individuals and private organizations, mainly from outside the mainstreams of the biomedical research community.⁸² The early laboratory research on chemical contraception described in this paper was but one important outcome of the concerted effort made by reformers in the 1920s to eliminate a variety of social problems thought to derive from excessive fertility. Scientific arguments and expertise were employed to advocate reform as well as to define the appropriate solution to such social problems. Scientists were recruited as advocates for the movement, but they were also employed as researchers in laboratory investigations sponsored by these same reformers.Sponsors of these early laboratory studies noted the difficulty of obtaining first-class investigators.⁸³ The routine analyses necessary for such research, as well as the traditional scientific aversion to applied problems, provide only a partial explanation for this response. The real difficulty lay in recruiting investigators to a field (reproduction and human sexuality) that had previously been taboo. Once opened up — first as socially relevant, and finally as scientifically sound — there was much interest in this area, and the appeal to researchers of the scientific issues surrounding fertility and reproduction soon surpassed that of the reforming value of birth control.A survey of the kinds of experimental investigations sponsored by birth control advocates indicates the range of physiological problems explored by contraceptive research. The most definitive work was done on the efficacy and safety of spermicides, but the potential of other contraceptive methods was also examined. Investigators attempted to develop spermatoxins that would effectively immunize women against sperm, and they also tried to elucidate the mechanism of hormonal control of reproduction. In fact, speculations about the possible hormonal manipulation of fertility were expressed at the Seventh International Birth Control Conference held in Zurich in 1930.⁸⁴ In the 1920s, clinical studies were undertaken to assess the effectiveness of the various birth control methods. Laboratory investigators complemented this work by screening spermicides for safety and testing for their ability to kill sperm. There were a variety of birth control preparations on the market (most of which were sold as feminine hygiene products), but no one really knew whether these were effective or even safe.⁸⁵ Although the physiology of other major organ systems was well advanced, the scientific study of reproductive physiology and contraceptive technology was clearly in its infancy in this period. Routine analyses simply could not be conducted, because the fundamental research establishing baselines had not yet been done. Scientists used this fact to redirect attention to basic research on reproduction.Laboratory research on contraception indicated important unexplored areas for physiological investigation. Social activists, who had encouraged prominent scientists to become interested in both the social value and the genetic implications of birth control, found these investigators revising the goals of their research. The biologists had formed their own network and had begun to seek out funding, reformulating the justification for sponsorship of further investigations. The eugenic motivations underlying these studies, which had initially made them theoretically attractive to biologists, were gradually eroded. Concern with human evolution ceded its place to interest in physiological mechanisms. Crew and others began to note that the use of biological theory to justify essentially political decisions had serious limitations. Biologists had become uncomfortable with those very arguments which had originally captured their interest. Recognition of the potential political abuse wrought by applying scientific principles to society was expressed by Crew just one year after the Zurich meeting. Referring to previous assessments of the role of sex in reproduction, he generalized: In the past the biologist has justified feudalism, Manchester Liberalism, socialism and every other type of social organization and political programme by reference to selected biological phenomena.⁸⁶ By 1932 Crew had also begun to question the biological logic of regulated breeding, and had made it clear to his American sponsor that there was no simple correspondence between the practice of birth control and the genetic improvement of the human race.⁸⁷ Biologists further began to recognize, however, that although the hopeful genetic solution to human problems was probably an illusion, contraception still remained one tangible means to alleviate, human misery. Some laboratory scientists, like Crew, acknowledged the applicability of their own particular skills to this problem. For a few brief years, social needs and scientific goals were mutually supportive and closely intertwined. But as laboratory researchers gained interest in the study of reproduction and established their own priorities in this field, they temporarily withdrew from the arena of debate over birth control as an important mechanism for social reform.With the rise of Hitler, the genetic arguments for birth control rapidly lost their appeal. But by that time the scientific problem of how to achieve effective contraception had entered the professional consciousness. Both physicians and scientists began to be aware of birth control as a subject within their domain of expertise, although outside the principal focus of their research. Scientific discussion of birth control permanently altered from a question of justification to a problem of method: How could one achieve reliable and safe contraception? This had been Sanger's and Dickinson's goal from the beginning. Laboratory scientists had indeed been persuaded to undertake this work; this research had in turn affected biologists' perceptions of the whole field of reproductive physiology, encouraging further study of reproductive mechanisms. The promise of new knowledge provided for continued funding of this research, despite the caution by scientists that the social benefits would not be as immediate or as far-reaching as advocates and they themselves had first argued.The activities of birth control activists and their supporting agencies, and the financial backing of private contributors and foundations, notably the Rockefeller philanthropies, provided an important new stimulus to the development of research on the biology of reproduction in the late 1920s and early 1930s. Biologists were able to claim an enlarged realm of issues for scientific study through their activities as advocates and as investigators for the birth control movement. At the same time, they promised as-yet-undiscovered possibilities for regulating human reproduction once its physiology was understood. The knowledge and control that they promised lay in understanding the whole reproductive cycle — not merely in evaluating the toxic effects of presumptive spermicides.Chemical spermicides never summoned the interest of scientists as the contraceptive pill was to do, yet that research did reinforce the widespread perceptions of scientific research as essential to social reform. Spermicide investigations focused research efforts in reproductive biology by challenging traditional taboos, defining problems for further study, and providing laboratory investigators the opportunity to assert the social and scientific value of their own skills. Crew echoed this attitude as he observed in 1934:Man has turned from the adventurous conquest of his environment to the conquest of himself. To-day is the day of biological invention, eagerly used for the control of the undesirable and the unwanted. Sex and reproduction are no longer hedged around by myth and taboo; they are no longer accepted as mysteries that defy understanding. They are matters inviting examination and explanation; they are regarded as expressions of physico-chemical forces, the nature of which is to be displayed. It is accepted that when knowledge is sufficient, control will be absolute, and, though knowledge is not yet sufficient, readers ... must be persuaded to the view that this will not always be so.⁸⁸ The synergism between reproductive biology and social needs has been temporary and sporadic, but recurring, since the 1920s. Scientific research programs have clearly been influenced by issues raised by public debate. Nevertheless, reproductive biologists have continued to assert their own professional goals. For the most part, they have rejected problems without inherent scientific interest and have spurned applied research except as it has had a direct bearing on current research themes. This attitude, apparent among American and British investigators in the 1930s, created the intellectual context for the invention and acceptance of the technically sophisticated oral contraceptive pill. It did not foster the production and improvement of the simple, safe, effective, and cheap vaginal contraceptive desired by early birth control advocates.     

Strontium stratigraphy of the Oligocene–Early Miocene shellbeds of the Kutch Basin,western India,and its implications

The Kutch Basin is unique among the western Indian sedimentary basins because of its near-complete sequence of post-Palaeozoic rocks. Due to extensive marine influence, the Oligocene–Early Miocene formations of the basin, namely Maniyara Fort, Khari Nadi and Chhasra, contain numerous shellbeds. Although age assignments of these formations exist based on foraminiferal biostratigraphy, detailed numerical age of the lithounits are yet to be established. We have identified a total of eleven distinct shellbeds (oldest SB 01 to youngest SB11) from this interval primarily containing bivalve fossils. Using ⁸⁷Sr/⁸⁶Sr of selected oyster and pectinid shells with pristine shell characteristics, we report the age of four shellbeds. The ages of SB 01, SB 04, SB 06 and SB 10 are 24.37, 17.31, 16.85 and 15.38 Ma, respectively. Our dates suggest a Chattian (24.37 Ma) age for SB 01 from the Bermoti Member, validating the previous biostratigraphical estimates from the Maniyara Fort Formation. The Chhasra Formation, however, shows a younger range of ages (17.31–15.38 Ma) characterized by a transition from the Burdigalian (SB 04–SB 06) to the Langhian (SB 10) stages. These dates have important implications in the study of sequence stratigraphy, Palaeobiogeography and tectonic history of the Kutch Basin. A surface with subaerial exposure is found in SB 08 (between 16.85 and 15.38 Ma) that corresponds to a global eustatic sea-level decrease (Mi2). Our new dates will also help evaluate the response of marine fauna to the closure of the Tethyan seaway around 19 Ma due to the formation of ‘Gomphotherium Landbridge’. The dated shellbeds enable us to identify pre- and post-closure fauna and assess the effect of biogeographical separation on these fauna. These dates have important implications in evaluating the regional geological record of western India in the context of various global events.     

Peptide, Peptidomimetic, and Small-molecule Antagonists of the p53–HDM2 Protein–Protein Interaction

Modulation of intracellular protein–protein interactions has been – and remains – a challenging goal for the discovery and development of small-molecule therapeutic agents. Progress in the pharmacological targeting and understanding at the molecular level of one such interaction that is relevant to cancer drug research, viz. that between the tumour suppressor protein p53 and its negative regulator HDM2, is reviewed here. The first X-ray crystal structure of a complex between a small peptide from the trans-activation domain of p53 and the N-terminal domain of HDM2 was reported almost 10 years ago. The nature of this interaction, which involves just three residue side chains in the p53 peptide ligand and a compact hydrophobic binding pocket in the HDM2 receptor, together with the attractive concept of reactivating the anti-proliferative functions of p53 in tumour cells, has spurned a great deal of effort aimed at finding drug-like antagonists of this interaction. A variety of approaches, including both structure-guided peptidomimetic and de novo design, as well as high through-put screening campaigns, have provided a wealth of leads that might be turned into actual drugs. There is still some way to go as far as optimisation and preclinical development of such leads is concerned, but it is clear already now that antagonists of the p53–HDM2 protein–protein interaction have a good chance of ultimately being successful in providing a new anti-cancer therapy modality, both in monotherapy and to potentiate the effectiveness of existing chemotherapies.     

Thermal, mixing, and oxygen regimes of the Salton Sea, California, 1997–1999

The Salton Sea is a shallow (mean depth = 8 m; maximum depth = 15 m), saline (41–45 g l^–1), intermittently mixing, 57 km long, 980 km² lake located in the arid southwestern United States. The Sea is a wind driven system, with predominant winds paralleling the long axis of the lake, being strongest in spring and weakest in summer and fall. The Sea mixed daily or nearly daily between September and January. During this cooling period, moderate to high levels of dissolved oxygen (3–11 mg l^–1) were found throughout the water column. Mean water column temperature ranged from a minimum of 13–14 °C in early January to a maximum of 30–34 °C in July–September. During most of this warming period, the Sea was thermally stratified but subject to periodic wind driven mixing events. Winds were stronger in spring 1998 than in 1997 or 1999, causing more rapid heating of the lake that year and also delaying onset of anoxic conditions in bottom waters. During summer months, mid-lake surface waters were sometimes supersatured with oxygen, and bottom waters were hypoxic or anoxic with sulfide concentrations > 5 mg l^–1. Oxic conditions (> 1 mg O₂ l^–1) often extended a few meters deeper nearshore than they did well offshore as a consequence of greater mixing nearshore. Mixing events in late summer deoxygenated the entire water column for a period of days. Consumption of oxygen by sulfide oxidation likely was the principal mechanism for these deoxygenation events. Sulfide concentrations in surface waters were 0.5–1 mg l^–1 approximately 3 days after one mixing event in mid-August 1999. These mixing events were associated with population crashes of phytoplankters and zooplankters and with large fish kills. In the southern basin, freshwater inflows tended to move out over the surface of the Sea mixing with saline lake water as a function of wind conditions. Salinity gradients often contributed more to water column stability than did thermal gradients in the southeasternmost portion of the lake.     

The Relative Significance of Host–Habitat,Depth, and Geography on the Ecology,Endemism, and Speciation of Coral Endosymbionts in the Genus Symbiodinium

Dinoflagellates in the genus Symbiodinium are among the most abundant and important group of eukaryotic microbes found in coral reef ecosystems. Recent analyses conducted on various host cnidarians indicated that Symbiodinium assemblages in the Caribbean Sea are genetically and ecologically diverse. In order to further characterize this diversity and identify processes important to its origins, samples from six orders of Cnidaria comprising 45 genera were collected from reef habitats around Barbados (eastern Caribbean) and from the Mesoamerican barrier reef off the coast of Belize (western Caribbean). Fingerprinting of the ribosomal internal transcribed spacer 2 identified 62 genetically different Symbiodinium. Additional analyses of clade B Symbiodinium using microsatellite flanker sequences unequivocally characterized divergent lineages, or “species,” within what was previously thought to be a single entity (B1 or B184). In contrast to the Indo-Pacific where host-generalist symbionts dominate many coral communities, partner specificity in the Caribbean is relatively high and is influenced little by the host’s apparent mode of symbiont acquisition. Habitat depth (ambient light) and geographic isolation appeared to influence the bathymetric zonation and regional distribution for most of the Symbiodinium spp. characterized. Approximately 80% of Symbiodinium types were endemic to either the eastern or western Caribbean and 40–50% were distributed to compatible hosts living in shallow, high-irradiance, or deep, low-irradiance environments. These ecologic, geographic, and phylogenetic patterns indicate that most of the present Symbiodinium diversity probably originated from adaptive radiations driven by ecological specialization in separate Caribbean regions during the Pliocene and Pleistocene periods.     

Evolution,palaeoecology, and palaeobiogeography of the Late Ordovician–Early Silurian brachiopod Epitomyonia

Epitomyonia is characterized by various types of dorsal ridges, which may be transverse, longitudinal, or highly convoluted and probably served as skeletal supports for lophophores of various complexity. Multivariate analyses suggest that the Epitomyonia-bearing brachiopod associations lived in relatively shallow-water environment in the Late Ordovician, and inhabited mainly deep-water environments in the early Wenlock. The temporal and spatial change in the faunal distribution may be explained by three alternative scenarios: (1) Epitomyonia followed the broad evolutionary trend of the Palaeozoic Evolutionary Fauna to shift from shallow- to deeper-water settings over time; (2) the dicoelosiid communities could not compete with the large-shelled pentameride communities in continental shelf settings during the Early Silurian; or (3) only the shallow-water Epitomyonia died out in the Late Ordovician mass extinction event, whereas some poorly known deep-water Late Ordovician forms survived into the Early Silurian. Epitomyonia paucitropida n. sp. from the lower Whittaker Formation (late Katian) of the Mackenzie Mountains, northwestern Canada, is reported as the first known Ordovician species of Epitomyonia from the palaeocontinent of Laurentia, characterized by a small shell with weak, transverse dorsal ridges that are most primitive for the genus.     

The carbohydrate–polypeptide linkages, the amino acid sequences of the peptides adjacent to some of these bonds, and the composition and size of the carbohydrate units of α1-acid glycoprotein

1. The glycopeptides derived from a proteolytic digest of sialic acid-free α₁-acid glycoprotein were separated on a DEAE-cellulose column into five main fractions. 2. The average molecular weight of these glycopeptides was 2400, except for one fraction whose molecular weight was 3100. The average molecular weight of the sialic acid-free carbohydrate units was found to be 2200. From these data and the carbohydrate content of the native protein and the assumed molecular weight of 44000, it was concluded that α₁-acid glycoprotein probably possesses five carbohydrate units. The sialic acid-containing carbohydrate units of this glycoprotein have an average molecular weight of 3000, except for one unit the molecular weight of which is significantly higher. 3. The N-, non-N- and C-terminal amino acids of the main glycopeptides were determined. Aspartic acid and threonine occur in most peptides. Alanine, glycine, proline, serine and lysine were present in varying amounts. Traces of other amino acids were also found. 4. The amino acid sequence of three main glycopeptides was established and indicated that these glycopeptides are located at different positions of the polypeptide chain of the glycoprotein. These sequences are: Asp(NH₂)-Pro-Lys; Thr-Asp(NH₂)-Ala; Asp(NH₂)-Gly-Thr. 5. From the results of a series of chemical reactions (periodate oxidation, hydrazinolysis, dinitrophenylation, mild acid hydrolysis) it was shown that the hydroxyl group of the N-terminal threonine and the -amino group of lysine are free and that the β-carboxyl group of aspartic acid is present as amide. It was concluded that this amide group is involved in the carbohydrate–polypeptide linkages of at least four carbohydrate units of α₁-acid glycoprotein. 6. The carbohydrate composition of the sialic acid-free glycopeptides was determined in terms of moles of neutral hexoses, glucosamine and fucose/mole. 7. Fucose, at least to the larger part, is not linked to sialic acid, and its (glycosidic) linkage is significantly more stable toward acid hydrolysis than the bond of the sialyl residues. 8. Heterogeneity of the carbohydrate units of α₁-acid glycoprotein was found with regard to size and to content of fucose and sialic acid.     

Travel costs, oviposition behaviour and the dynamics of insect–plant systems

Insect attack can have major consequences for plant population dynamics. We used individually based simulation models to ask how insect oviposition behaviour influences persistence and potential stability of an herbivore–plant system. We emphasised effects on system dynamics of herbivore travel costs and of two kinds of behaviour that might evolve to mitigate travel costs: insect clutch size behaviour (whether eggs are laid singly or in groups) and female aggregation behaviour (whether females prefer or avoid plants already bearing eggs). Travel costs that increase as plant populations drop lead to inverse density dependence of plant reproduction under herbivore attack. Female clutch size and aggregation behaviours also strongly affect system dynamics. When females lay eggs in large clutches or aggregate their clutches, herbivore damage varies strongly among plants, providing probabilistic refuges that permit plant reproduction and persistence. However, the population dynamics depend strongly on whether insect behaviour is fixed or responds adaptively to plant population size: when (and only when) females increase clutch size or aggregation as plants become rare, refuges from herbivory weaken at high plant density, creating inverse density dependence in plant reproduction. Both herbivore travel costs themselves, and also insect behaviour that might evolve in response to travel costs, can thus create plant density dependence—a basic requirement for regulation of plant populations by their insect herbivores.     

Detection,isolation, and characterization of chikungunya viruses associated with the Pakistan outbreak of 2016–2017

The chikungunya virus(CHIKV) is a mosquito-transmitted alphavirus, which has infected millions of people in Africa, Asia, Americas, and Europe since it reemerged in India and Indian Ocean regions in 2005–2006. Starting in the middle of November 2016, CHIKV has been widely spread, and more than 4,000 cases of infections in humans were confirmed in Pakistan. Here, we report the first isolation and characterization of CHIKV from the Pakistan outbreak. Eight CHIKV strains were newly isolated from human serum samples using a cell culture procedure. A full-length genome sequence and eight complete envelope(E1) sequences of CHIKV from Pakistan were obtained in this study. Alignment of the CHIKV E1 sequences revealed that the eight new CHIKV isolates were highly homogeneous, with only two nonsynonymous substitutions found at generally conserved sites(E99 and Q235). Based on the comparison of 342 E1 sequences, the two nonsynonymous mutations were located in well-recognized domains associated with viral functions such as the cell fusion and vector specificity, suggesting their potential functional importance. Phylogenetic analysis indicated that the CHIKV strains from Pakistan originated from CHIKV circulating in the Indian region. This study helps elucidate the epidemics of CHIKV in Pakistan and also provides a foundation for studies of evolution and expansion of CHIKV in South Asia.     

N-Methylated Analogs of hIAPP Fragments 18–22, 23–27, 33–37 Inhibit Aggregation of the Amyloidogenic Core of the Hormone

Amylin (hIAPP) aggregation leads to the formation of insoluble deposits and is one of the factors in the development of type II diabetes. The aim of this research was to find N-methylated analogs of the aggregating amylin fragments 18–22, 23–27, and 33–37, which would not themselves be susceptible to aggregation and would inhibit the aggregation of the amyloidogenic cores of the hormone. None of the analogs of fragment 18–22 containing one or two N-methylated amino acid residues showed any tendency to aggregate. Only the peptide H−F(N−Me)GA(N−Me) IL−OH ( 6 ) derived from the 23–27 hIAPP hot spot did not form fibrous structures. All analogs of the 33–37 amylin fragment were characterized by the ability to form aggregates, despite the presence of N-methylated amino acids in their structures. N-Methylated peptides 1 – 5 demonstrated inhibitory properties against the aggregation of fragment 18–22. Aggregation of the amyloidogenic core of 23–27 was significantly inhibited by N-methylated peptides 1 – 3 derived from the (18–22) H−HSSNN−OH fragment and by the H−F(N-Me)GA(N−Me)IL−OH ( 6 ) fragment derived from the 23–27 amylin hot spot. Fragment (33–37) H−GSNTY−NH₂ was found to be inhibited in the presence of N-methylated peptides 1 – 3 derived from the 18–22 fragment and by the double methylated peptide H−F(N−Me)GA(N−Me)IL−OH ( 6 ). Research on the possibility of using N-methylated analogs of amyloidogenic amylin cores as inhibitors of hormone aggregation is ongoing, with a focus on finding the minimum concentration of N-methylated peptides capable of inhibiting the aggregation of hIAPP hot spots.     

Characterisation of the thiol–disulphide chemistry of desmopressin by LC, μ-LC, LC-ESI-MS and Maldi-Tof

Summary. To date, the majority of therapeutic peptides and proteins have to be administered via parenteral routes, which are painful and inconvenient. In order to gain sufficient high blood concentrations after oral application, various barriers in the gastrointestinal tract have to be overcome. Apart from a poor membrane uptake and intense enzymatic degradation, this study will demonstrate that thiol–disulphide reactions are an underestimated essential part of the presystemic metabolism. Glutathione, integrative part of the antioxidant defence system in the gastrointestinal tract, may play an important role in the inactivation of orally given peptides and proteins. In order to verify this hypothesis, desmopressin which bears a single disulphide bond was used as model peptide drug. Desmopressin was incubated with GSH in various concentrations, and the extent of thiol/disulphide exchange reactions between the peptide drug and GSH was investigated in dependence on pH and ratio of reactants determined as a function of time via HPLC, LC-MS and Maldi-Tof-MS analyses. Results showed that desmopressin is degraded by 1% reduced glutathione at pH 4 and pH 5.5. In presence of 0.01%, 0.1% and 1% of reduced glutathione 6.1%, 19.4% and 52.1% of desmopressin, respectively, were degraded. The masses of the conjugates after deconvolution measured by liquid chromatography and electrospray ionisation mass spectrometric detection were m/z 1069.67, m/z 1376.50, m/z 1683.40 and m/z 2138. These molecular masses, confirmed by Maldi-Tof-MS analysis, correspond with the masses of conjugates expected in theory. Under defined conditions, these results reveal that thiol–disulphide exchange reactions have a considerable impact on the alteration of peptide drugs and proteins.     

Review of the fauna of Rotatoria,Cladocera, and Copepoda of the basin of the Anadyr’ River

The zooplankton composition is studied in the thermokarst, glacial and meteorite lakes, channels, former riverbeds, and hollows in the basin of Anadyr’. We found 174 taxa: 78, Rotatoria, 55, Cladocera, and 41, Copepoda. The most diverse is the lake fauna: 51 taxa of Rotatoria, 48, Cladocera, and 37, Copepoda. The thermokarst Lake Maiorskoe hosts 68 taxa: 31, Rotatoria, 14, Cladocera, and 23, Copepoda, wheras the cold ultraoligotrophic Lake El’gygytgyn features only one species of Cyclop of the group scutifer Cyclops neymanae Strel., and Rotatoria and Cladocera are present as allochtonous forms. The Copepoda illustrate the relations of the Anadyr’ fauna with those of Europe, North America, and Japan.