Synthesis,carbonic anhydrase I and II inhibition studies of the 1,3,5-trisubstituted-pyrazolines

4-(3-(4-Substituted-phenyl)-5-phenyl-4,5-dihydro-1H-pyrazol-1-yl) benzenesulfonamides (9–16) were successfully synthesized and their chemical structures were confirmed by ¹H NMR, ¹³C NMR, and HRMS spectra. Carbonic anhydrase I and II inhibitory effects of the compounds were investigated. K_i values of the compounds were in the range of 316.7?±?9.6–533.1?±?187.8?nM towards hCA I and 412.5?±?115.4–624.6?±?168.2?nM towards hCA II isoenzymes. While K_i values of the reference compound Acetazolamide were 278.8?±?44.3?nM and 293.4?±?46.4?nM towards hCA I and hCA II izoenzymes, respectively. Compound 14 with bromine and compound 13 with fluorine substituents can be considered as the leader compounds of the series because of the lowest K_i values in series to make further detailed carbonic anhydrase inhibiton studies.     

Synthesis and biological evaluation of aminomethyl and alkoxymethyl derivatives as carbonic anhydrase,acetylcholinesterase and butyrylcholinesterase inhibitors

Compounds containing nitrogen and sulfur atoms can be widely used in various fields such as industry, medicine, biotechnology and chemical technology. Therefore, the reactions of aminomethylation and alkoxymethylation of mercaptobenzothiazole, mercaptobenzoxazole and 2-aminothiazole were developed. Additionally, the alkoxymethyl derivatives of mercaptobenzoxazole and 2-aminothiazole were synthesized by a reaction with hemiformals, which are prepared by the reaction of alcohols and formaldehyde. In this study, the inhibitory effects of these molecules were investigated against acetylcholinesterase (AChE), butyrylcholinesterase (BChE) enzymes and carbonic anhydrase I, and II isoenzymes (hCA I and II). Both hCA isoenzymes were significantly inhibited by the recently synthesized molecules, with K_i values in the range of 58–157?nM for hCA I, and 81–215?nM for hCA II. Additionally, the K_i parameters of these molecules for BChE and AChE were calculated in the ranges 23–88 and 18–78?nM, respectively.     

Rhesus macaque KIR bind human MHC class I with broad specificity and recognize HLA-C more effectively than HLA-A and HLA-B

Human killer cell immunoglobulin-like receptors (KIR) recognize A3/11, Bw4, C1, and C2 epitopes carried by mutually exclusive subsets of human leukocyte antigen (HLA)-A, -B, and -C allotypes. Chimpanzee and orangutan have counterparts to HLA-A, -B, and -C, and KIR that recognize the A3/11, Bw4, C1, and C2 epitopes, either individually or in combination. Because rhesus macaque has counterparts of HLA-A and -B, but not HLA-C, we expected that rhesus KIR would better recognize HLA-A and -B, than HLA-C. Comparison of the interactions of nine rhesus KIR3D with 95 HLA isoforms, showed the KIR have broad specificity for HLA-A, -B, and -C, but vary in avidity. Considering both the strength and breadth of reaction, HLA-C was the major target for rhesus KIR, followed by HLA-B, then HLA-A. Strong reactions with HLA-A were restricted to the minority of allotypes carrying the Bw4 epitope, whereas strong reactions with HLA-B partitioned between allotypes having and lacking Bw4. Contrasting to HLA-A and -B, every HLA-C allotype bound to the nine rhesus KIR. Sequence comparison of high- and low-binding HLA allotypes revealed the importance of polymorphism in the helix of the α₁ domain and the peptide-binding pockets. At peptide position 9, nonpolar residues favor binding to rhesus KIR, whereas charged residues do not. Contrary to expectation, rhesus KIR bind more effectively to HLA-C, than to HLA-A and -B. This property is consistent with major histocompatibility complex (MHC)-C having evolved in hominids to be a generally superior ligand for KIR than MHC-A and MHC-B.     

The adaptor function of SHP-2 downstream of the prolactin receptor is required for the recruitment of p29, a substrate of SHP-2

SHP-2, a cytosolic protein tyrosine phosphatase with two SH2 domains and multiple tyrosine phosphorylation sites, contributes to signal transduction as an enzyme and/or adaptor molecule. Here we demonstrate that prolactin (PRL) stimulation of the PRL-responsive Nb2 cells, a rat lymphoma cell line, and T47D cells, a human breast cancer cell line, lead to the complex formation of SHP-2 and growth factor receptor-bound protein-2 (grb2). Using transient co-overexpression studies of the prolactin receptor (PRLR) and several tyrosine to phenylalanine mutants of SHP-2, we show that grb2 associates with SHP-2 through the C-terminal tyrosine residues of SHP-2, Y(546) and Y(584). Furthermore, in this study, we found a highly phosphorylated, 29-kDa protein (p29), a substrate of SHP-2. The recruitment of p29 to SHP-2 requires the carboxy-terminal tyrosine residues of SHP-2 (Y(546) and Y(584)). Together, our results indicate that SHP-2 may function as an adaptor molecule downstream of the PRLR and highlight a new recruitment mechanism of SHP-2 substrates.     

Lithostratigraphy and carbonate microfacies across the Permian–Triassic boundary near Julfa (NW Iran) and in the Baghuk Mountains (Central Iran)

Permian–Triassic boundary sections in the Julfa (NW Iran) and Abadeh (Central Iran) regions display a succession of three characteristic rock units, (1) the Paratirolites Limestone with the mass extinction horizon at its top, (2) the ‘Boundary Clay’, and (3) the earliest Triassic Elikah Formation with the conodont P–Tr boundary at its base. The carbonate microfacies reveals a facies change, in the sections near Julfa, within the Paratirolites Limestone with an increasing number of intraclasts, Fe–Mn crusts, and biogenic encrustation. A decline in carbonate accumulation occurs towards the top of the unit with a sponge packstone in the sections, and finally resulting in a complete demise of the carbonate factory. The succession of the ‘Boundary Clay’ differs in the two regions; thin horizons of sponge packstone are present in the Julfa region and ‘calcite fans’ of probably inorganic origin in the Abadeh Region. The skeletal carbonate factory of the Late Permian was restored with the deposition of microbial carbonates at the base of the Elikah Formation, where densely laminated bindstone, floatstone with sparry calcite spheres, and oncoid wackestone/floatstone predominate.     

PDCD10 Gene Mutations in Multiple Cerebral Cavernous Malformations

Cerebral cavernous malformations (CCMs) are vascular abnormalities that may cause seizures, intracerebral haemorrhages, and focal neurological deficits. Familial form shows an autosomal dominant pattern of inheritance with incomplete penetrance and variable clinical expression. Three genes have been identified causing familial CCM: KRIT1/CCM1, MGC4607/CCM2, and PDCD10/CCM3. Aim of this study is to report additional PDCD10/CCM3 families poorly described so far which account for 10-15% of hereditary cerebral cavernous malformations. Our group investigated 87 consecutive Italian affected individuals (i.e. positive Magnetic Resonance Imaging) with multiple/familial CCM through direct sequencing and Multiplex Ligation-Dependent Probe Amplification (MLPA) analysis. We identified mutations in over 97.7% of cases, and PDCD10/CCM3 accounts for 13.1%. PDCD10/CCM3 molecular screening revealed four already known mutations and four novel ones. The mutated patients show an earlier onset of clinical manifestations as compared to CCM1/CCM2 mutated patients. The study of further families carrying mutations in PDCD10/CCM3 may help define a possible correlation between genotype and phenotype; an accurate clinical follow up of the subjects would help define more precisely whether mutations in PDCD10/CCM3 lead to a characteristic phenotype.     

Environmental Profile of Brazilian Green Coffee (6 pp)

Goal, Scope and Background

Brazil is the world's biggest producer of coffee beans with approx. a 30% market share. Depending on climate conditions, approx. 30 million bags of coffee beans are exported annually from Brazil, while domestic consumption is around 10 million bags, which makes Brazil the world's third largest coffee-consuming country. Therefore, the goal of this paper is to present the LCA of green coffee produced in Brazil for the reference crops 2001/02 and 2002/03 in order to generate detailed production inventory data as well as to identify the potential environmental impacts of its tillage in order to realize how to reduce those impacts and increase the environmental sustainability of this product. Only the inputs and outputs relative to the coffee tillage were considered. The production of fertilizers, correctives and pesticides were not included in the boundary, but only their amounts. The functional unit selected for this study was 1,000 kg of green coffee destined for exportation.

Methods

The LCI was performed according to the ISO 14040 standard series. All information considered in this study (use of water, fossil based energy, fertilizers and chemicals) were taken up in in-depth data collection and evaluation by questionnaires applied on a farm level and/or received by mail. Four Brazilian coffee producer regions were evaluated: Cerrado Mineiro, South of Minas Gerais State, the Marília and Alta Mogiana regions in São Paulo State. These regions have the following geographic coordinates: 44 to 50° W longitude and 18 to 24° S latitude. The data refer to a production of 420,000 coffee bean bags and a productive area of approx. 14,300 ha. The varieties of coffee beans considered in this study were Mundo Novo, Catuaí (yellow and red), Icatu (yellow and red), Catucaí (yellow and red) and Obatã. Farm specific data along with agricultural production data have been combined to elaborate a coffee cultivation inventory, which will be applied in an emissions estimation.

Results and Conclusion

The production of 1,000 kg of green coffee in Brazil requires approx. 11,400 kg of water, 94 kg of diesel, 270 kg of fertilizers as NPK, 900 kg of total fertilizers, 620 kg of correctives, 10 kg of pesticides and 0.05 hectare of annual land use. Outputs related to these functional units are approx. 3,000 kg of waste water from coffee washing, 8,500 kg of waste water from the wet method and 750 kg of organic residue that is reincorporated to the tillage as fertilizer. The publication of an LCI of agricultural products is a fundamental step for understanding the potential environmental impacts of each tillage and then establishes the basis for product sustainability. In this way, this work is the first Brazilian initiative for applying LCA to coffee cultivation.

Recommendation and Perspective

Different agricultural practices demonstrate different environmental profiles. The amount of agricultural pesticide is directly related to agricultural practices as tillage rotation, density of plants, etc. This study supplied important results for a better correlation of the agricultural practices and potential environmental impacts of coffee. Future updates of this study will show the evolution of the natural resource management such as land use, new agricultural practices, lower fertilizers and chemicals use.