Enzyme Inhibitory and Anti-cancer Properties of Moringa peregrina / Propiedades inhibidoras Enzimáticas y Anticancerígenas de la Moringa peregrina

Background: Moringa peregrina is widely used in the traditional medicine of the Arabian Peninsula to treat various ailments, because it has many pharmacologically active components with several therapeutic effects. Objective: This study aimed to investigate the inhibitory effect of Moringaperegrina seed ethanolic extract (MPSE) against key enzymes involved in human pathologies, such as angiogenesis (thymidine phosphorylase), diabetes (α-glucosidase), and idiopathic intracranial hypertension (carbonic anhydrase). In addition, the anticancer properties were tested against the SH-SY5Y (human neuroblastoma). Results: MPSE extract significantly inhibited α-glucosidase, thymidine phosphorylase, and carbonic anhydrase with half-maximal inhibitory concentrations (IC50) values of 303.1 ± 1.3, 471.30 ± 0.3, and 271.30 ± 5.1 µg/mL, respectively. Furthermore, the antiproliferative effect of the MPSE was observed on the SH-SY5Y cancer cell line with IC50 values of 55.1 µg/mL. Conclusions: MPSE has interesting inhibitory capacities against key enzymes and human neuroblastoma cancer cell line.

Antecedentes: La Moringa peregrina se utiliza ampliamente en la medicina tradicional de la Península Arábiga para tratar diversas dolencias, ya que posee numerosos componentes farmacológicamente activos con varios efectos terapéuticos. Objetivo: Este estudio tenía como objetivo investigar el efecto inhibidor del extracto etanólico de semillas de Moringaperegrina (MPSE) frente a enzimas clave implicadas en patologías humanas, como la angiogénesis (timidina fosforilasa), la diabetes (α-glucosidasa) y la hipertensión intracraneal idiopática (anhidrasa carbónica). Además, se comprobaron las propiedades anticancerígenas frente al SH-SY5Y (neuroblastoma humano). Resultados: El extracto de MPSE inhibió significativamente la α-glucosidasa, la timidina fosforilasa y la anhidrasa carbónica con concentraciones inhibitorias semimáximas (IC50) de 303,1 ± 1,3, 471,30 ± 0,3 y 271,30 ± 5,1 µg/mL, respectivamente. Además, se observó el efecto antiproliferativo del MPSE en la línea celular del cáncer SH-SY5Y con valores de IC50 de 55,1 µg/mL. Conclusiones: MPSE posee interesantes capacidades inhibitorias frente a enzimas clave y línea celular de neuroblastoma canceroso humano.

Carbonic anhydrase IX-based tumor imaging and therapy: a review / 生物工程学报

Carbonic anhydrase IX (CAIX) is a transmembrane protein that is specifically overexpressed on the surface of hypoxic tumor cells. With the function of regulating the acidity of tumor cells both inside and outside, CAIX is closely related to tumor proliferation, invasion and metastasis. Therefore, CAIX is a promising target for tumor imaging and therapy. Herein, we summarized recent advances in CAIX-based tumor imaging, therapy and theranostics, and prospected future applications of using CAIX as an anti-tumor target.

miR-7/TGF-β2 axis sustains acidic tumor microenvironment-induced lung cancer metastasis

Acidosis, regardless of hypoxia involvement, is recognized as a chronic and harsh tumor microenvironment (TME) that educates malignant cells to thrive and metastasize. Although overwhelming evidence supports an acidic environment as a driver or ubiquitous hallmark of cancer progression, the unrevealed core mechanisms underlying the direct effect of acidification on tumorigenesis have hindered the discovery of novel therapeutic targets and clinical therapy. Here, chemical-induced and transgenic mouse models for colon, liver and lung cancer were established, respectively. miR-7 and TGF-β2 expressions were examined in clinical tissues (n = 184). RNA-seq, miRNA-seq, proteomics, biosynthesis analyses and functional studies were performed to validate the mechanisms involved in the acidic TME-induced lung cancer metastasis. Our data show that lung cancer is sensitive to the increased acidification of TME, and acidic TME-induced lung cancer metastasis via inhibition of miR-7-5p. TGF-β2 is a direct target of miR-7-5p. The reduced expression of miR-7-5p subsequently increases the expression of TGF-β2 which enhances the metastatic potential of the lung cancer. Indeed, overexpression of miR-7-5p reduces the acidic pH-enhanced lung cancer metastasis. Furthermore, the human lung tumor samples also show a reduced miR-7-5p expression but an elevated level of activated TGF-β2; the expressions of both miR-7-5p and TGF-β2 are correlated with patients' survival. We are the first to identify the role of the miR-7/TGF-β2 axis in acidic pH-enhanced lung cancer metastasis. Our study not only delineates how acidification directly affects tumorigenesis, but also suggests miR-7 is a novel reliable biomarker for acidic TME and a novel therapeutic target for non-small cell lung cancer (NSCLC) treatment. Our study opens an avenue to explore the pH-sensitive subcellular components as novel therapeutic targets for cancer treatment.

Advances of enzymes related to microbial cement / 生物工程学报

Microbial induced calcium carbonate precipitation (MICP) refers to the natural biological process of calcium carbonate precipitation induced by microbial metabolism in its surrounding environment. Based on the principles of MICP, microbial cement has been developed and has received widespread attention in the field of biology, civil engineering, and environment owing to the merits of environmental friendliness and economic competence. Urease and carbonic anhydrase are the key enzymes closely related to microbial cement. This review summarizes the genes, protein structures, regulatory mechanisms, engineering strains and mutual synergistic relationship of these two enzymes. The application of bioinformatics and synthetic biology is expected to develop biocement with a wide range of environmental adaptability and high performance, and will bring the MICP research to a new height.

Quantitative Characterization of the Chemical Space Governed by Human Carbonic Anhydrases and selenium-containing derivatives of solfonamides

Abstract Due to the fact that different isoforms of carbonic anhydrase play distinct physiological roles, their diseases/disorders involvement are different as well. Involvement in major disorders such as glaucoma, epilepsy, Alzheimer's disease, obesity and cancers, have turned carbonic anhydrase into a popular case study in the field of rational drug design. Since carbonic anhydrases are highly similar with regard to their structures, selective inhibition of different isoforms has been a significant challenge. By applying a proteochemometrics approach, herein the chemical interaction space governed by acyl selenoureido benzensulfonamides and human carbonic anhydrases is explored. To assess the validity, robustness and predictivity power of the proteochemometrics model, a diverse set of validation methods was used. The final model is shown to provide valuable structural information that can be considered for new selective inhibitors design. Using the supplied information and to show the applicability of the constructed model, new compounds were designed. Monitoring of selectivity ratios of new designs shows very promising results with regard to their selectivity for a specific isoform of carbonic anhydrase.

Silencing carbonic anhydrase 1 inhibits the proliferation, invasion and migration of A549 cells in lung cancer / 中国临床药理学与治疗学

AIM: To investigate the effects of silencing carbonic anhydrase 1 (CA1) on proliferation, apoptosis, invasion and migration of human lung cancer A549 cells. METHODS: CA1-specific siRNA (si-CA1 group) and negative control (si-NC group) were transfected into lung cancer A549 cells by lipofection. The A549 cells transfected with empty liposome were used as blank control group. Real-time quantitative PCR (qPCR) and Western blot (Western blot) were used to detect the expression of CA1 mRNA and protein. Cell counting kit method (CCK-8), flow cytometry and Transwell assay were used to detect proliferation and apoptosis of A549 cells, invasion and migration capabilities. RESULTS:qPCR and Western blot showed that the expression levels of CA1 mRNA and protein in A549 cells transfected with CA1 siRNA were significantly down-regulated (P0.05). CONCLUSION: Silencing CA1 can inhibit the proliferation, invasion and migration of lung cancer A549 cells and promote cell apoptosis.

Enzyme inhibition and antibacterial potential of 4-Hydroxycoumarin derivatives

The 4-Hydroxycoumarin derivatives are known to show a broad spectrum of pharmacological applications. In this paper we are reporting the synthesis of a new series of 4-Hydroxycoumarin derivatives synthesized through Knovenegal condensation; they were characterized by using UV-Vis, FT-IR, NMR spectroscopies. The synthesized compounds were evaluated for antibacterial activity against Staphylococcus aureus and Salmonella typhimurium strains. The compounds (2), (3) and (8) showed favorable antibacterial activity with zone of inhibitions 26.5± 0.84, 26.0 ± 0.56 and 26.0 ± 0.26 against Staphylococcus aureus (Gram-positive) respectively. However, the compounds (5) and (9) were found more active with 19.5 ± 0.59 and 19.5 ± 0.32 zone of inhibitions against Salmonella typhimurium (Gram-negative). Whereas, in urease inhibition assay, none of the synthesized derivatives showed significant anti-urease activity; although, in carbonic anhydrase-II inhibition assay, the compound (2) and (6) showed enzyme inhibition activity with IC50 values 263±0.3 and 456±0.1, respectively.

Inhibition of amyloid fibrillation of apo-carbonic anhydrase by flavonoid compounds

Flavonoids are polyphenol compounds abundantly found in plants and reported to have an inhibitory effect on amyloidfibrillation. The number and position of hydroxyl groups, as well as the arrangement of flavonoids rings, may influencetheir inhibitory effects. In this study, we investigate the effect of structural characteristics of flavonoids on amyloid fibrilformation. For this purpose, five compounds (i.e., biochanin A, daidzein, quercetin, chrysin and fisetin) were selected thatrepresent a variety in the number and position of their hydroxyl groups. The inhibitory effect of these flavonoids on theamyloid fibril formation of apo-carbonic anhydrase (apo-BCA), as a model protein, was evaluated using thioflavin T andtransmission electron microscopy. The results showed that fisetin possessed the most significant inhibitory effect. Interestingly, upon apo-BCA acetylation, none of the tested flavonoids could inhibit the fibrillation process, which indicates thatthe interactions of these compounds with the amine groups of lysine residues could be somewhat important

Mining and engineering of microbial carbonic anhydrases for biomimetic carbon dioxide sequestration / 生物工程学报

The increasing atmospheric carbon dioxide levels have been correlated with global warming. Carbonic anhydrases (CA) are the fastest among the known enzymes to improve carbon capture. The capture of carbon dioxide needs high temperature and alkaline condition, which is necessary for CaCO₃ precipitation in the mineralization process. In order to use CAs for biomimetic carbon sequestration, thermo-alkali-stable CAs are, therefore, essential, and polyextremophilic microbes are one of the important sources of these enzymes. The current review focuses on both those isolated by thermophilic organisms from the extreme environments and those obtained by protein engineering techniques, and the industrial application of the immobilized CAs is also briefly addressed. To reduce the greenhouse effect and delay global warming, we think further research efforts should be devoted to broadening the scope of searching for carbonic anhydrase, modifying the technology of protein engineering and developing highly efficient immobilization strategies.

Research advances in hydrogen sulfide donors / 中国药科大学学报

@#Hydrogen sulfide(H2S)is an endogenous gas messenger molecule with extremely broad biological activities including vasodilation, anti-oxidation, anti-inflammation, cardioprotection and anti-tumor. Similar to other gas messenger molecules, the biological activity of H2S is dependent on its location, concentration and duration of exposure. Therefore, the key scientific issue is how to improve the selectivity of H2S donor molecules to release appropriate concentrations of H2S at the target site(commonly pathological place), exerting therapeutic efficacy with limited side-effects. This article reviews the structures and H2S release mechanisms of two classes of H2S donors focusing on the advances in the recently developed H2S donors with controllable release potential of H2S, thus providing new ideas for future H2S-based drug research.

Carbonic anhydrase IX immunohistochemistry has potential to predict renal cell carcinoma outcomes: A systematic review and meta-analyses

@#Introduction: Tissue biomarker carbonic anhydrase IX (CAIX) is purported to have prognostic value for renal cell carcinoma (RCC) but contradicting findings from previous studies have also been documented. This study aims to perform a systematic review and meta-analysis on the role of CAIX in RCC disease progression. Materials and Methods: Following the preferred reporting items for systematic review and meta-analysis (PRISMA) guidelines, online searches of multiple databases were performed to retrieve articles from their inception until December 2017. Inclusion criteria included all English-based original articles of immunohistochemistry (IHC) studies investigating CAIX expression in human RCC tissue. Four articles were finally selected for meta-analysis with a total of 1964 patients. Standard meta-analysis methods were applied to evaluate the role of CAIX in RCC prognosis. The relative risk (RR) and its 95% confidence interval (CI) were recorded for the association between biomarker and prognosis, and data were analysed using MedCalc statistical software. Results: The meta-analysis showed that high CAIX expression was associated with low tumour stage (RR 0.90%, 95% CI 0.849-0.969, p= 0.004), low tumour grade (RR 0.835%, 95% CI 0.732-0.983, p= 0.028), absence of nodal involvement (RR 0.814%, 95% CI 0.712-0.931, p= 0.003) and better ECOS-PS index (RR 0.888%, 95% CI 0.818-0.969, p= 0.007). The high tissue CAIX expression in RCC is hence an indication of an early malignancy with a potential to predict favourable disease progression and outcome. Conclusion: The measurement of this marker may be beneficial to determine the course of the illness. It is hoped that CAIX can be developed as a specific tissue biomarker for RCC in the near future

Aspirin interrupts bile duct carcinoma in rats induced by thioacetamide / 第二军医大学学报

Objective To establish a thioacetamide (TAA)-induced bile duct carcinoma rat model and to observe the effect of aspirin on bile duct carcinoma by interrupting the model. Methods Sprague-Dawley (SD) male rats were given drinking water containing TAA (300 mg/L) for 5 months, and tumor formations of the livers were observed by H-E staining at the 8th, 12th, 16th, and 20th weeks, respectively. The expression of carbonic anhydrase 2 (CA-2) in liver tissues was also detected by immunohistochemistry. SD male rats were also selected and treated with aspirin after exposing to TAA for 12 weeks. And then the tumor formations of the livers were observed by H-E staining at the 3rd and 6th months, and the expression of CA-2 was detected by immunohistochemistry. Bile duct carcinoma cells QBC939 were obtained and treated with aspirin (0 mmol/L and 5 mmol/L), and the expression of CA-2 was detected by Western blotting after culturing for 48 h. Results Obvious fibrosis was found in the livers of some rats at 12th weeks after exposing to TAA; large number of fibrous tissue hyperplasia and microscopic or suspected tumors were found in the livers at the 16th weeks; visible tumors in the livers were found in all the rats at the 20th weeks. The incidence of hepatic tumors or suspected tumors was 28.6% (2/7) in rats treated with aspirin for 6 months, while it was 100% (7/7) in rats without aspirin treatment. The expression of CA-2 in liver tissues was gradually increased with the development of bile duct carcinoma in rats, while the expression of CA-2 in liver tissues was decreased after treating with aspirin. The expression of CA-2 in QBC939 cells treated with 5 mmol/L aspirin for 48 h was significantly decreased versus the untreated control group (P0.01). Conclusion TAA can successfully induce intrahepatic bile duct carcinoma. Aspirin can interrupt the development of bile duct carcinoma and may be used to prevent intrahepatic bile duct carcinoma.

Establishment and application evaluation of the ELISA method for anti-carbonic anhydrase Ⅱ antibody / 国际检验医学杂志

Objective To establish an ELISA detection method for human serum carbonic anhydrase(CA)Ⅱ antibody,and to evaluate the level of serum CA Ⅱ antibody in patients with hypertensive nephropathy, chronic glomerulonephritis,type 2 diabetic nephropathy and healthy people.Methods To establish the ELISA method using CA Ⅱ,anti-CA Ⅱ monoclonal antibody and enzyme labeled secondary antibody,the evaluation of the degree of precision and sensitivity,and stability and anti-interference performance of the ELISA were made.Results The detection accuracy of serum CA Ⅱ antibody ELISA was 6.0%,the precision between the batches was 8.6%,the sensitivity was 0.032,and with favorable anti-interference performance and stability. The level of serum anti-CA Ⅱ antibody was significantly higher in patients with chronic glomerulonephritis and 2-type diabetic nephropathy compared with the normal control(P<0.05).There was no significant differ-ence in serum CAⅡ antibody level between hypertensive nephropathy group and healthy control group(P>0.05).Conclusion It is feasible to establish a serum CAⅡ antibody ELISA method to meet the needs of clini-cal test.CAⅡ antibody may be used as autoantibody to participate in the development of chronic glomerulone-phritis and type 2 diabetic nephropathy.

Different expression patterns of carbonic anhydrase IX in oral lichen planus and leukoplakia

Tumor hypoxia is an important indicator of cancer prognosis. Among the different genes that are upregulated by hypoxia is carbonic anhydrase IX, which combines carbon dioxide and water to form bicarbonate and hydrogen. Although expression of this enzyme is very low in normal tissues, carbonic anhydrase IX is overexpressed in several types of cancer. The aim of the present work was to analyze carbonic anhydrase IX expression in the two most frequent potentially malignant oral disorders: oral lichen planus and oral leukoplakia. Immunohistochemical analysis of oral lichen planus and oral leukoplakia biopsies was performed using anticarbonic anhydrase IX antibody. Samples of normal mucosa served as controls. Statistical analysis was performed by Fischer's exact test. The enzyme was detected in the epithelium of both lesions. The staining was more intense in the basal layer and decreased towards the surface in oral lichen planus. Conversely, the most intense reaction was observed in the superficial layers in leukoplakia, and staining intensity decreased towards the basal membrane. No carbonic anhydrase IX expression was seen in normal mucosa samples. Carbon anhydrase IX expression in lichen and leukoplakia epithelia shows that hypoxia plays a role in the pathogenesis of both lesions. The different distribution patterns provides further evidence of the different biological behavior of these two entities, which under certain circumstances can have similar clinical and histological features (AU)

La hipoxia tumoral es un importante indicador de pronóstico en cáncer. Entre los distintos genes que son activados por hipoxia, uno de los principales es la anhidrasa carbónica IX (CAIX), que combina CO2 con H2O para sintetizar HCO3 y H+. Aunque la expresión de esta enzima es muy baja en tejidos normales, se sobreexpresa en varios tipos de cáncer. La finalidad del presente trabajo fue analizar la expresión de CAIX en las dos lesiones orales potencialmente malignas más frecuentes: el liquen plano y la leucoplasia. Se utilizó una técnica inmuno histoquímica con un anticuerpo específico contra CAIX, en biopsias de liquen plano oral y leucoplasia oral. Se utilizaron mucosas normales como controles. Se realizaron análisis estadísticos utilizando test exacto de Fischer. La identificación de la enzima fue positiva en el epitelio de ambas lesiones. En los líquenes la reacción es más intensa en los estratos basales, disminuyendo hacia la superficie. Inversamente, las leucoplasias mostraron marcación más intensa en estratos superficiales, con disminución hacia la membrana basal. Las mucosas normales resultaron negativas. La expresión de CAIX en el epitelio de líquenes y leucoplasias indica que la hipoxia juega algún papel en la patogenia de ambas lesiones. El diferente patrón de distribución es una evidencia más del diferente comportamiento biológico de dos entidades las cuales en ciertas circunstancias pueden manifestar cuadros clínicos e histológicos semejantes (AU)

Hemoglobinuria paroxística nocturna. el rol de la anhidrasa carbónica-i urinaria en la hemólisis intravascular / Peroxysmal nocturnal hemoglobinuria: the role of urinary carbonic anhydrase-I in intravascular haemolysis

La Hemoglobinuria Paroxística Nocturna (HPN) se caracteriza por hemólisis intravascular crónica mediada por complemento. Cuando se produce la hemolisis se libera a circulación Anhidrasa Carbónica- I (AC-I), una enzima que se halla en alta concentración en el eritrocito y por su bajo peso molecular filtra por el glomérulo. El objetivo del presente trabajo fue detectar la excreción de la AC-I en orina de pacientes con HPN por Electroforesis Bidimensional de Utilidad Clínica (2D UC), y compararla con otras causas de hemólisis, de origen renal y postrenal. Se evaluaron 8 pacientes con HPN sin tratamiento con eculizumab un inhibidor del C5 del complemento, y 5 de ellos postratamiento, 12 orinas de pacientes con nefritis lúpica y 10 orinas de pacientes con hemólisis postrenal. La AC-I puede estar presente en la orina, en los tres grupos, sin embargo la relación AC-I/Hemoglobina en la hemólisis intravascular está invertida en comparación con la hemolisis glomerular y post-renal. Los pacientes con HPN tratados con eculizumab no presentan AC-I, y sería de utilidad en el seguimiento de los pacientes tratados con el inhibidor del C5, para evidenciar posibles escapes hemolíticos.

Paroxysmal Nocturnal Hemoglobinuria (PNH) is characterized by chronic complement mediated haemolysis. In these conditions it might be expected that carbonic anhydrase-I (AC-I) would be liberated into the plasma and excreted in the urine, by its high concentration in the erythrocyte and low molecular weight. The objective of the present study was to detect the urinary excretion of AC-I from patients with PNH by wodimensional clinical utility electrophoresis (2D UC) and to compare it with other causes of renal and post-renal haemolysis. We evaluated 8 patients with PNH without eculizumab, a complement C5 inhibitor, 5 of them posttreatment, 12 urine of patients with lupus nephritis and 10 urine of patients with post-renal hemolysis. AC-I may be present in the urine, in all three groups, however, the AC-I/Haemoglobin ratio in intravascular haemolysis is reversed compared to glomerular and post-renal haemolysis. Patients with PNH treated with eculizumab do not have AC-I and would be useful in monitoring patients treated with the C5 inhibitor to evidence possible haemolytic leaks.

Effect of methazolamide in patients with refractory uveitic macular edema / 国际眼科杂志(Guoji Yanke Zazhi)

AIM:To evaluate the efficacy and safety of methazolamide in treating refractory uveitic macular edema.METHODS: Retrospective self-controlled study was designed.A total of 15 patients (20 eyes) with refractory uveitic macular edema which used methazolamide as adjuvant therapy were enrolled in Shanghai First People`s Hospital from January 2015 to June 2016.The changes of central macular thickness (CMT) and best corrected visual acuity (BCVA) were observed at baseline and 2, 4, 8wk after treatment.We also focused on the incidence of complications and relapse.RESULTS: The CMT was 445.95±154.10μm, 338.83±138.34μm, 251.50±40.20μm, 244.90±35.68μm at baseline, 2, 4 and 8wk after treatment, respectively.The differences among them were statistically significant (F=15.467, P<0.05).The BCVA (log MAR) were 0.40±0.17, 0.28±0.21, 0.19±0.20, 0.18±0.21 at baseline, 2, 4 and 8wk respectively, with a significant difference among them (F=5.208, P<0.05).When the cumulative dose reached to 700mg and 1400mg, no one had methazolamide-related complications;and when it came to 2800mg, 5 patients (33%) had methazolamide-related complication.After the withdrawal of methazolamide 1wk, 1 and 3mo, 3 patients (20%), 5 patients (33%) and 8 patients (53%) relapsed, respectively.CONCLUSION: Methazolamide is beneficial in improving macular edema and vision in 4wk.When the cumulative dose is more than 1400mg, we need pay attention to the complications.After discontinuing methazolamide for 1wk, macular edema relapsed in some patients, and more than half of patients recurred after 3mo.So the patients should be followed closely in 3mo after withdrawal of methazolamide.

Bioassay-guided isolation of novel and selective urease inhibitors from Diospyros lotus / 中国天然药物

Two new dimeric naphthoquinones, 5',8'-dihydroxy-6,6'-dimethyl-7,3'-binaphthyl-1,4,1',4'-tetraone (1; Di-naphthodiospyrol D) and 5',8'-dihydroxy-5,8-dimethoxy-6,6'-dimethyl-7,3'-binaphthyl-1,4,1',4'-tetraone (2; Di-naphthodiospyrol E), along with known naphthoquinones diospyrin (3) and 8-hydroxy diospyrin (4) were isolated from the chloroform fraction of extract of Diospyros lotus roots. Their structures were elucidated by advanced spectroscopic analyses, including HSQC, HMBC, NOESY, and J-resolved NMR experiments. The fractions and compounds 1-4 were evaluated for urease activity and phosphodiesterase-I, carbonic anhydrase-II and α-chymotrypsin enzyme inhibitory activities. Compounds 1 and 2 and their corresponding fractions showed significant and selective inhibitory effects on urease activities. The IC values of 1 and 2 were 260.4 ± 6.37 and 381.4 ± 4.80 µmol·L, respectively, using thiourea (IC = 21 ± 0.11 µmol·L) as the standard inhibitor. This was the first report demonstrating that the naphthoquinones class showed urease inhibition.

Effects of dorzolamide/timolol and tafluprost on intraocular pressure and pupil diameter in healthy dogs / Efeitos da dorzolamida/timolol e da tafluprosta sobre a pressão intraocular e o diâmetro pupilar em cães saudáveis

ABSTRACT: This study aimed to evaluate and compare the effects of the fixed combination of dorzolamide/timolol with those of tafluprost on intraocular pressure (IOP) and pupil diameter (PD) in healthy dogs (n=10). Two experiments were conducted with an interval of 30 days. In both, IOP and PD were assessed at 8, 11, 14, 17, and 20h. Parameters were evaluated during baseline, treatment period of four days, and one day of post-treatment. During treatment phase, IOP decreased by 0.74 (P<0.05), 1.88 (P<0.01), 2.94 (P<0.001), and 3.10mmHg (P<0.01), in dorzolamide/timolol-treated eyes; and by 1.50, 2.18, 2.14, and 2.18mmHg (P<0.001), in tafluprost-treated eyes. PD decreased by 0.24 (P<0.01), 0.32 (P<0.01), 0.49 (P<0.001), and 0.40mm (P<0.001), in dorzolamide/timolol treated eyes; and by 2.31, 2.55, 2.43, and 2.70mm (P<0.001), in tafluprost-treated eyes. Dorzolamide/timolol and tafluprost were able to decrease IOP and PD in healthy dogs. However, a cumulative effect of the fixed combination of dorzolamide/timolol was more effective in reducing IOP, than tafluprost. Comparisons between treatments showed that tafluprost was more effective in reducing PD throughout the treatment phase.

RESUMO: O estudo objetivou avaliar e comparar os efeitos da combinação fixa da dorzolamida/timolol com os da tafluprosta sobre a pressão intraocular (PIO) e o diâmetro pupilar (DP) em cães saudáveis (n=10). Dois experimentos com intervalo de 30 dias foram conduzidos. Em ambos, a PIO e o DP foram avaliados às 8, 11, 14, 17 e às 20h. Os parâmetros foram avaliados durante a fases basal, um período de tratamento de quatro dias, e um dia de pós-tratamento. Durante a fase de tratamento, a PIO dos olhos tratados com dorzolamida/timolol reduziram em 0.74 (P<0.05), 1.88 (P<0.01), 2.94 (P<0.001), e 3.10mmHg (P<0.01); e dos olhos tratados com tafluprosta em 1.50, 2.18, 2.14 e 2.18mmHg (P<0.001). O DP dos olhos tratados com dorzolamida/timolol reduziram em 0.24 (P<0.01), 0.32 (P<0.01), 0.49 (P<0.001) e 0.40mm (P<0.001); e dos olhos tratados com tafluprosta em 2.31, 2.55, 2.43 e 2.70mm (P<0.001). A dorzolamida/timol e a tafluprosta foram capazes de reduzir a PIO e o DP em cães saudáveis. Porém, efeito cumulativo do tratamento com dorzolamida/timolol foi observado, decorridos três dias de tratamento. Por essa razão, a dorzolamida/timolol foi mais efetiva que a tafluprosta na redução da PIO. Comparações entre os tratamentos demonstraram que a tafluprosta foi mais efetiva em reduzir o DP, durante toda a fase de tratamento.

Carbonic Anhydrase Inhibition Affect Antioxidant Enzyme Activity in the Blood of STZ Induced Diabetic Rats.

Background: Carbonic anhydrase is found in the blood of all vertebrate and thus playing a fundamental role in the maintenance of acid-base homeostasis. Erythrocytes are intrinsically prone to oxidative stress because of their exposure to high oxygen tension. Aim: The study aimed to investigate the changes of erythrocytes anti-oxidative enzymes in STZ induced diabetic rats and to determine the antioxidant potential of Cadaba farinosa leaves. Results: The result of the present study showed that inhibition of carbonic anhydrase result in significant decrease in both erythrocyte and plasma catalase activity, whereas erythrocyte and plasma superoxide dismutase activity increased. Conclusion: Carbonic anhydrase inhibition may alter the activity of anti-oxidative enzymes in vivo.

The Expression of Carbonic Anhydrase (CA) IX/XII and Lymph Node Metastasis in Early Breast Cancer / Journal of the Korean Cancer Association, 대한암학회지

PURPOSE: The aim of study was to test by immunohistochemical (IHC) staining whether carbonic anhydrase (CA) 9 and 12 have an effect on sentinel lymph node (SLN) metastasis in early breast cancer and to find clinicopathologic factors associated with SLN metastasis. MATERIALS AND METHODS: Between June 2003 and June 2011, medical records of 470 patients diagnosedwith breast cancer with pT1-2, pN0-2, and M0 were reviewed. Of these 470, 314 patients who underwent SLN biopsy+/-axillary dissection were subjects of this study. Using tissue microarray, IHC staining for CA9 and CA12 was performed. Clinicopathologic factors such as patient age, tumour size, lymphatic invasion, hormone receptor status, and the Ki-67 labeling index were analysed together. RESULTS: The mean age of all patients was 51.7 years. The mean number of harvested SLN was 3.62, and 212 patients (67.5%) had negative SLN. Lymphatic invasion, the Ki-67 labelling index of primary tumours, and CA9 staining of stromal cells, were independent risk factors for SLN metastasis in the multivariate analysis. In 33 patients (10.5%) without the three risk factors, no patient had SLN metastasis. In 80 patients without lymphatic invasion of primary tumours or CA9 staining of stromal cells, only four patients (5%) had positive SLN. CONCLUSION: CA9 staining of stromal cells is an independent risk factor for SLN metastasis as well as lymphatic invasion and a low Ki-67 labelling index of primary tumours in patients with early breast cancer. IHC staining of primary tumours for CA12was not associatedwith SLN metastasis.