Synthesis and biological activity of anticoagulant heparan sulfate glycopolymers

Heparin has been used as an anticoagulant drug for more than 70 years. The global distribution of contaminated heparin in 2007, which resulted in adverse clinical effects and over 100 deaths, emphasizes the necessity for safer alternatives to animal-sourced heparin. The structural complexity and heterogeneity of animal-sourced heparin not only impedes safe access to these biologically active molecules, but also hinders investigations on the significance of structural constituents at a molecular level. Efficient methods for preparing new synthetic heparins with targeted biological activity are necessary not only to ensure clinical safety, but to optimize derivative design to minimize potential side effects. Low molecular weight heparins have become a reliable alternative to heparin, due to their predictable dosages, long half-lives, and reduced side effects. However, heparin oligosaccharide synthesis is a challenging endeavor due to the necessity for complex protecting group manipulation and stereoselective glycosidic linkage chemistry, which often result in lengthy synthetic routes and low yields. Recently, chemoenzymatic syntheses have produced targeted ultralow molecular weight heparins with high-efficiency, but continue to be restricted by the substrate specificities of enzymes.

To address the need for access to homogeneous, complex glycosaminoglycan structures, we have synthesized novel heparan sulfate glycopolymers with well-defined carbohydrate structures and tunable chain length through ring-opening metathesis polymerization chemistry. These polymers recapitulate the key features of anticoagulant heparan sulfate by displaying the sulfation pattern responsible for heparin’s anticoagulant activity. The use of polymerization chemistry greatly simplifies the synthesis of complex glycosaminoglycan structures, providing a facile method to generate homogeneous macromolecules with tunable biological and chemical properties. Through the use of in vitro chromogenic substrate assays and ex vivo clotting assays, we found that the HS glycopolymers exhibited anticoagulant activity in a sulfation pattern and length-dependent manner. Compared to heparin standards, our short polymers did not display any activity. However, our longer polymers were able to incorporate in vitro and ex vivo characteristics of both low-molecular-weight heparin derivatives and heparin, displaying hybrid anticoagulant properties. These studies emphasize the significance of sulfation pattern specificity in specific carbohydrate-protein interactions, and demonstrate the effectiveness of multivalent molecules in recapitulating the activity of natural polysaccharides.

Tunable heparan sulfate glycomimetics for modulating chemokine activity

Heparan sulfate (HS) glycosaminoglycans participate in critical biological processes by modulating the activity of a diverse set of protein binding partners. Such proteins include all known members of the chemokine superfamily, which are thought to guide the migration of distinct subsets of immune cells through their interactions with HS proteoglycans on endothelial cell surfaces. Animal-derived heparin polysaccharides have been shown to reduce inflammation levels through the inhibition of HS-chemokine interactions; however, the clinical usage of heparin as an anti-inflammatory drug is hampered by its anticoagulant activity and potential risk for side effects, such as heparin-induced thrombocytopenia (HIT).

Here, we describe an expedient, divergent synthesis to prepare defined glycomimetics of HS that recapitulate the macromolecular structure and biological activity of natural HS glycosaminoglycans. Our synthetic approach uses a core disaccharide precursor to generate a library of four differentially sulfated polymers. We show that a trisulfated glycopolymer antagonizes the chemotactic activities of pro-inflammatory chemokine RANTES with similar potency as heparin polysaccharide, without potentiating the anticoagulant activities of antithrombin III. The same glycopolymer also inhibited the homeostatic chemokine SDF-1 with significantly more efficacy than heparin. Our work offers a general strategy for modulating chemokines and dissecting the pleiotropic functions of HS/heparin through the presentation of defined sulfation motifs within multivalent polymeric scaffolds.

Insight on how fishing bats discern prey and adjust their mechanic and sensorial features during the attack sequence

Several insectivorous bats have included fish in their diet, yet little is known about the processes underlying this trophic shift. We performed three field experiments with wild fishing bats to address how they manage to discern fish from insects and adapt their hunting technique to capture fish. We show that bats react only to targets protruding above the water and discern fish from insects based on prey disappearance patterns. Stationary fish trigger short and shallow dips and a terminal echolocation pattern with an important component of the narrowband and low frequency calls. When the fish disappears during the attack process, bats regulate their attack increasing the number of broadband and high frequency calls in the last phase of the echolocation as well as by lengthening and deepening their dips. These adjustments may allow bats to obtain more valuable sensorial information and to perform dips adjusted to the level of uncertainty on the location of the submerged prey. The observed ultrafast regulation may be essential for enabling fishing to become cost-effective in bats, and demonstrates the ability of bats to rapidly modify and synchronise their sensorial and motor features as a response to last minute stimulus variations.

The effect of organotin and copper sulfate on the late development and presettlement behavior of the hard clam Mercenaria mercenaria

The effect of organotin compounds and copper, commonly used as antifouling agent, were studied on Mercenaria mercernaria larvae. They were reared under usual hatchery conditions until they reached 190 um in diameter. The larvae were subjected to four compounds, tributylin chloride (TBT), monobutyltin chloride (MBT), trimethyltin chloride (TMT), cupric sulfate (CuSo4) plus control. Mortality was measured at 24, 48 h, and 96h. Behavioral and/or metamorphic changes were recorded in triplicate at 24-48 and 96 h. The appearance in swimming larvae of a functional foot was considered a sign of competence to set and was recorded as a "pediveliger". Swimming larvae were considered as larvae that have not yet reached their total development and they were recorded as "swimming". Larvae that did not show foot or swimming activity and were static but alive on the bottom were recorded as "bottom". TBT was found to completely inhibit swimming activity at sublethal concentrations throughout the period of observation. Copper and MBT inhibited swimming from 48 h, TMT did not inhibit swimming activity at any of the times recorded. The four compounds ranked in order of decreasing toxicity were TBT>TMT>CU>MBT.

On acid sulfate soils of the coastal aquaculture ponds of Bangladesh

With the stimulus of the very high international market value of penaeid shrimp, new pond areas for shrimp farming are rapidly being added in Bangladesh. Unfortunately, this expansion is occurring with the loss of some natural mangrove forests and with soils and sediments that are far from ideal for aquaculture. In this study, two representative shrimp farming areas were surveyed and pH, in profile depth, was recorded. It was found that the shrimp farming areas of the Chakaria Sundarban are more acidic than those of the Khulna-Satkhira region due to the acid sulfate soils.

Studies on arsenical creosote as a wood preservative for marine structures. Part 2. Observation on leaching, corrosion and resistance to borer attack

A detailed study on arsenical creosote with reference to leaching, corrosion and anti-borer properties was carried out. Results showed that aging had very little effect on the preservative which suggested better fixation of the preservative into the wood. Corrosion of mild steel, galvanised iron, aluminium-magnesium alloy (M57S) and copper panels in the preservative was found to be negligible. Normal creosote and low temperature creosote of Regional Research Laboratory, Hyderabad, both fortified with arsenic trioxide resisted borer damage on wooden panels for a period of over five months in the port of Cochin. The performance of low temperature creosote fortified with arsenic was found to be equally satisfactory when compared to normal creosote fortified in the same manner. A loading of 208.6 Kgs/ml³ for Haldu (Adina cordifolia) and 138 Kgs/m³ for Mango (Mangifera indica) in the case of normal creosote and 177 Kgs/m³ for Mango the case of RRL creosote were found to be sufficient for treating the wood.

Tolerance of Penaeus monodon larvae to cupric sulfate added in bath

Copper is used to deter the growth of bacterial, fungal and protozoan disease organism in fishes. Zoeae (Z SUB-1 ), myses (M SUB-1 ) and postlarvae (P SUB-1 ) were exposed to copper sulfate at concentrations of 0 . 025, 0 . 05, 0 . 75, 0 . 1 and 0 . 2 ppm from 24 to 96 hours. The number of surviving larvae were counted at the end of each 24-hour period and the percentage of survival is determined for each dose level. The LC SUB-50 for each of the larval stages was interpolated from the data whenever possible. Three trials with 2 replicates per trial were conducted. The physico-chemical characteristics of the bath taken before and at the end of the experimental period show insignificant differences between initial and final values in each trial. Results indicate that mortality rates of all larval stages increased with exposure time and that mortality rates of the experimental group is higher than the control. Interpolation of the LC SUB-50 is possible only for the 48-h and 72-h exposure times for both zoeae and myses and for the 48-h exposure time for the postlarvae. This is due to the high survival percentage of the 24-h group and the low survival percentage (below 50%) of the larvae exposed for 96 hours. The 48-hour LC SUB-50 for Z SUB-1 , M SUB-1 and P SUB-1 are 0 . 225, 0 . 350 and 0 . 125 ppm respectively. Postlarvae seem to be more sensitive than either of the 2 larval stages having a lower 48-h LC SUB-50 and a low survival rate after 72 hours. The larvae were observed to lose their balance and were lethargic, producing few swimming movements so that they were mostly confined to the bottom of the aquaria. Moribund larvae observed under the microscope had a faster but weak heartbeat compared to healthy larvae. Slight or complete loss of feeding ability indicated by empty guts and delayed molting of Z SUB-1 to Z SUB-2 were also noted.

Halichondria sulfonic acid, a new HIV-1 inhibitory guanidino-sulflonic acid, and halistanol sulfate isolated from the marine sponge Halichondria rugosa Ridley & Dendy

A new sulfur-containing guanidino derivative, halichondria sulfonic acid (1) showing anti-HIV-1 activity, and halistanol trisulfate (2) with anti-tumor activity have been isolated from the marine sponge Halichondria rugosa Ridley & Dendy collected in the