The present study evaluated in-vitro toxicity of organophosphate insecticide, malathion on nuclea... more The present study evaluated in-vitro toxicity of organophosphate insecticide, malathion on nuclear maturation, fertilization and subsequent embryo development of the oocytes obtained from abattoir ovaries of Nili-Ravi Buffalo (Bubalus bubalis). The cumulus-oocyte complexes (COCs) in culture medium were exposed to four different regimens of malathion, 1.56 µmol, 3.12 µmol, 6.25 µmol and 12.5 µmol. Results demonstrated dose dependent significant difference (P < 0.05) in nuclear competence, oocyte maturation and degeneration. Cumulus expansion showed significant increase (P < 0.05) while oocyte diameter increased non significantly (P > 0.05). The COCs that attained nuclear maturation (MII) and were subjected to IVF and, subsequent embryonic development also showed dose dependent non-significant (P > 0.05) decrease in fertilization and embryonic development, however, hatched blastocyst formation was significantly compromised (P < 0.05). The blastocyst development with inn...
SUMMARY The effects of equilibration times (E1, 2 h; E2, 4 h; E3, 6 h), freezing rates (FR1, manu... more SUMMARY The effects of equilibration times (E1, 2 h; E2, 4 h; E3, 6 h), freezing rates (FR1, manual, 5 cm above liquid nitrogen (LN 2) for 10 min, plunging in LN 2 ; FR2, programmable ultra-fast, holding at +4 °C for 2 min, from 4 to À10 °C at À10 °C/min, from À10 to À20 °C at À15 °C/min, from À20 to À120 °C at À60 °C/min, holding at À120 °C for 30 sec, plunging in LN 2), and thawing rates (T1, 37 °C for 30 sec; T2, 50 °C for 15 sec; T3, 70 °C for 7 sec) were evaluated on quality of buffalo bull spermatozoa. Progressive motility (%), rapid velocity (%), average path velocity (VAP, lm/s), straight line velocity (VSL, lm/s), and mitochondrial transmembrane potential (%) were higher (p &lt; 0.05) with E2, FR2, and T3 compared to other groups. Sperm curved line velocity (VCL, lm/s) was higher (p &lt; 0.05) with E2 and FR2 compared to other groups. Sperm straightness (%) and linearity (LIN, %) were higher (p &lt; 0.05) with E2 compared to other groups. Sperm LIN was affected (p &lt; 0.05) with T3 compared to other groups. Supravital-plasma membrane integrity (%), viability and acrosome integrity (%) of spermatozoa were higher (p &lt; 0.05) with E2 and FR2 compared to other groups. Sperm DNA integrity (%) was higher (p &lt; 0.05) with FR2 and T1 compared to other groups. We concluded that inclusion of 4 h-equilibration time, programmable ultra-fast freezing rate, and rapid thawing at 70 °C for 7 sec in cryopreservation protocol improves the post-thaw quality of buffalo bull spermatozoa.
Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermato... more Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermatozoa. Semen from each of the five bulls (n = 3 replicates, six ejaculates/bull, a total of 30 ejaculates) was diluted in Tris-citric acid extender containing curcumin (0.5, 1.0, 1.5 or 2.0 mM) or control. At pre-freezing and post-thawing, total antioxidant contents (μM/L) and lipid peroxidation levels (μM/ml) were higher (p &lt; .05) and lower (p &lt; .05) respectively, with 1.5 and 2.0 mM compared to 0.5 and 1.0 mM curcumin and control. At post-thawing, progressive motility (PM, %) and rapid velocity (RV, %) were higher (p &lt; .05) with 1.5 mM compared to other doses of curcumin and control (except in case of RV, 1.5 was similar with 1.0 mM). Kinematics (average path velocity, μm/s; straight-line velocity, μm/s; curved-line velocity, μm/s; straightness, %; linearity, %), in vitro longevity (%, PM and RV) and DNA integrity (%) at post-thawing were higher (p &lt; .05) with 1.5 mM compared to control. At post-thawing, supravital plasma membrane integrity (%) and viable spermatozoa with intact acrosome (%) were higher with 1.5 compared to 2.0 mM curcumin and control. We concluded that freezability of water buffalo spermatozoa is improved with the addition of 1.5 mM curcumin in extender.
This study was primarily designed to evaluate the effect of different concentrations of ultraviol... more This study was primarily designed to evaluate the effect of different concentrations of ultraviolet (UV)-C-irradiated chicken egg yolk plasma (EYP; v:v; 10%, P1; 15%, P2; 20%, P3) or 20% (v:v) of whole chicken egg yolk (WCEY) in tris-citric acid (TCA) extender on water buffalo sperm quality during cryopreservation (postdilution, PD; postequilibration, PE; post-thawing, PT). Also the effect of best evolved concentration of UV-C-irradiated EYP in extender on in vivo fertility of buffalo spermatozoa was evaluated. At PE and PT, computer-assisted sperm analysis progressive motility (PM, %) was significantly higher in P3 compared with P1 and WCEY. Rapid velocity (RV, %) was higher (P < 0.05) in P3 compared with P1 and WCEY during cryopreservation (PD, PE, and PT). Average path velocity (μm/s) and straight line velocity (μm/s) were higher (P < 0.05) in P2 and P3 than WCEY at PE and PT. The decline percentage (%, longevity) in PM and RV was lower (P < 0.05) in P3 compared with WCE...
Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermato... more Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermatozoa. Semen from each of the five bulls (n = 3 replicates, six ejaculates/bull, a total of 30 ejaculates) was diluted in Tris-citric acid extender containing curcumin (0.5, 1.0, 1.5 or 2.0 mM) or control. At pre-freezing and post-thawing, total antioxidant contents (μM/L) and lipid peroxidation levels (μM/ml) were higher (p < .05) and lower (p < .05) respectively, with 1.5 and 2.0 mM compared to 0.5 and 1.0 mM curcumin and control. At post-thawing, progressive motility (PM, %) and rapid velocity (RV, %) were higher (p < .05) with 1.5 mM compared to other doses of curcumin and control (except in case of RV, 1.5 was similar with 1.0 mM). Kinematics (average path velocity, μm/s; straight-line velocity, μm/s; curved-line velocity, μm/s; straightness, %; linearity, %), in vitro longevity (%, PM and RV) and DNA integrity (%) at post-thawing were higher (p < .05) with 1.5 mM compared...
This study was primarily designed to evaluate the effect of different concentrations of ultraviol... more This study was primarily designed to evaluate the effect of different concentrations of ultraviolet (UV)-C-irradiated chicken egg yolk plasma (EYP; v:v; 10%, P1; 15%, P2; 20%, P3) or 20% (v:v) of whole chicken egg yolk (WCEY) in tris-citric acid (TCA) extender on water buffalo sperm quality during cryopreservation (postdilution, PD; postequilibration, PE; post-thawing, PT). Also the effect of best evolved concentration of UV-C-irradiated EYP in extender on in vivo fertility of buffalo spermatozoa was evaluated. At PE and PT, computer-assisted sperm analysis progressive motility (PM, %) was significantly higher in P3 compared with P1 and WCEY. Rapid velocity (RV, %) was higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P3 compared with P1 and WCEY during cryopreservation (PD, PE, and PT). Average path velocity (μm/s) and straight line velocity (μm/s) were higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P2 and P3 than WCEY at PE and PT. The decline percentage (%, longevity) in PM and RV was lower (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P3 compared with WCEY during 2 hours incubation under in vitro condition at PT. Supravital plasma membrane integrity (%) was higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P2 and P3 compared with control at different stages (PE and PT). Mitochondrial transmembrane potential (%) was higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P2 and P3 compared with P1 and WCEY at different stages (PD and PT). Percentage of viable sperm with intact acrosome, and sperm DNA integrity (%) were higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P2 and P3 compared with WCEY at PT. The in vivo fertility rate (%) was significantly higher with P3 compared with WCEY (76.61 vs. 64.49). In conclusion, WCEY (20%) can be replaced with UV-C-irradiated chicken EYP (20%) in TCA extender for cryopreservation of water buffalo spermatozoa.
The receptors for sulphonylurea (SURs) are known to be expressed in the mouse kidney, but their e... more The receptors for sulphonylurea (SURs) are known to be expressed in the mouse kidney, but their expression in the ureter is undefined. Owing to the physiological and pharmacological significance of SURs, the localization of SUR in ureters of adult mice and rats was investigated through immunohistochemistry. Animals were perfused transcardially with 4% paraformaldehyde and tissues were processed for immunohistochemistry using polyclonal antisera against SUR2A and SUR2B (SUR1) receptor proteins. Sections were incubated with primary and secondary antisera and developed with aminoethylcarbazole as a chromogen. A differentiated localized staining pattern of SUR proteins in rat and mouse ureters is demonstrated. In the mouse, immunoreactivity of SUR2A was predominantly confined to the cytoplasmic portion of epithelial cells and blood vessels, with comparatively low-level staining found in smooth muscle. In contrast, SUR2B (SUR1) immunoreactivity was absent in mouse ureters. In rats, SUR2A...
Pfl�gers Archiv European Journal of Physiology, 1999
The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glib... more The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glibenclamide, which are widely used as oral hypoglycaemic agents. Sulphonylureas have also been shown to affect urine flow and salt excretion by the kidney; therefore, the use of these drugs may have important implications for the pharmacological manipulation of renal salt handling. The purpose of the present investigation was to increase our understanding of the possible role of SUR in the regulation of renal function by determining the distribution of SUR isoforms within mouse kidney. Immunostaining with anti-SUR antisera revealed specific staining of SUR2B in distal nephron segments of mouse kidney. A diffuse, low level staining was observed in proximal tubules in the inner cortical region. No evidence was found for the presence of SUR2B in intra-renal blood vessels. Reverse-transcription polymerase chain reaction and Western blotting experiments indicated that SUR2B is the only known isoform expressed. These data demonstrate that SUR2B in mouse kidney is expressed in tubule regions that are critical in determining renal salt excretion.
The receptors for sulphonylurea (SURs) are known to be expressed in the mouse kidney, but their e... more The receptors for sulphonylurea (SURs) are known to be expressed in the mouse kidney, but their expression in the ureter is undefined. Owing to the physiological and pharmacological significance of SURs, the localization of SUR in ureters of adult mice and rats was investigated through immunohistochemistry. Animals were perfused transcardially with 4% paraformaldehyde and tissues were processed for immunohistochemistry using polyclonal antisera against SUR2A and SUR2B (SUR1) receptor proteins. Sections were incubated with primary and secondary antisera and developed with aminoethylcarbazole as a chromogen. A differentiated localized staining pattern of SUR proteins in rat and mouse ureters is demonstrated. In the mouse, immunoreactivity of SUR2A was predominantly confined to the cytoplasmic portion of epithelial cells and blood vessels, with comparatively low-level staining found in smooth muscle. In contrast, SUR2B (SUR1) immunoreactivity was absent in mouse ureters. In rats, SUR2A immunoreactivity was localized only in the blood vessels, while SUR2B (SUR1) immunoreactivity was localized in the epithelial cell cytoplasm. Tissue specificity of SUR is demonstrated in the two species of rodents and suggests a role of SUR proteins in urinary metabolism pertaining possibly to salt handling and maintenance of the smooth muscle tone.
Pflugers Archiv-European Journal of Physiology, 1999
The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glib... more The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glibenclamide, which are widely used as oral hypoglycaemic agents. Sulphonylureas have also been shown to affect urine flow and salt excretion by the kidney; therefore, the use of these drugs may have important implications for the pharmacological manipulation of renal salt handling. The purpose of the present investigation was to increase our understanding of the possible role of SUR in the regulation of renal function by determining the distribution of SUR isoforms within mouse kidney. Immunostaining with anti-SUR antisera revealed specific staining of SUR2B in distal nephron segments of mouse kidney. A diffuse, low level staining was observed in proximal tubules in the inner cortical region. No evidence was found for the presence of SUR2B in intra-renal blood vessels. Reverse-transcription polymerase chain reaction and Western blotting experiments indicated that SUR2B is the only known isoform expressed. These data demonstrate that SUR2B in mouse kidney is expressed in tubule regions that are critical in determining renal salt excretion.
The objective of the study was to devise a cryoprotection synergism between glycerol and dimethyl... more The objective of the study was to devise a cryoprotection synergism between glycerol and dimethyl sulfoxide (DMSO) for water buffalo sper-matozoa. Additionally, the effect of best evolved concentrations of glycerol and DMSO in extender was assessed on in vivo fertility of buffalo spermatozoa. Ejaculates (n = 30) were equally distributed into five aliquots; first aliquot was diluted at 37 °C in extender having 7 % glycerol (control); second aliquot was diluted at 37 °C as well as at 4 °C in extender having 3.5 % DMSO (Group 1); third aliquot was diluted at 37 °C in extender having 3.5 % glycerol and then at 4 °C in extender having 3.5 % DMSO (Group 2); fourth aliquot was diluted at 37 °C in extender having 3.5 % DMSO and then at 4 °C in extender having 3.5 % glycerol (Group 3); fifth aliquot was diluted in extenders having 1.75 % glycerol and 1.75 % DMSO at 37 as well as at 4 °C (Group 4). At post thawing, sperm progressive motility (%), rapid velocity (%), average path velocity (lm/s), curved line velocity (lm/s), in vitro longevity (%), structural and functional integrity of plasmalemma (%), mitochondrial transmembrane potential (%) and viable sperm with intact acrosome (%) were higher (P \ 0.05) in Group 4 compared to other treatment groups and control. Regarding sperm DNA integrity (%); it was higher (P \ 0.05) in Group 4 compared to Group 1, 3 and control. The in vivo fertility (%) of buffalo sperma-tozoa was significantly higher with Group 4 compared to control (69.45 vs. 59.81). In conclusion, synergism exists between glycerol and DMSO (Group 4) in improving the quality and in vivo fertility of cryop-reserved water buffalo spermatozoa.
The present study evaluated in-vitro toxicity of organophosphate insecticide, malathion on nuclea... more The present study evaluated in-vitro toxicity of organophosphate insecticide, malathion on nuclear maturation, fertilization and subsequent embryo development of the oocytes obtained from abattoir ovaries of Nili-Ravi Buffalo (Bubalus bubalis). The cumulus-oocyte complexes (COCs) in culture medium were exposed to four different regimens of malathion, 1.56 µmol, 3.12 µmol, 6.25 µmol and 12.5 µmol. Results demonstrated dose dependent significant difference (P < 0.05) in nuclear competence, oocyte maturation and degeneration. Cumulus expansion showed significant increase (P < 0.05) while oocyte diameter increased non significantly (P > 0.05). The COCs that attained nuclear maturation (MII) and were subjected to IVF and, subsequent embryonic development also showed dose dependent non-significant (P > 0.05) decrease in fertilization and embryonic development, however, hatched blastocyst formation was significantly compromised (P < 0.05). The blastocyst development with inn...
SUMMARY The effects of equilibration times (E1, 2 h; E2, 4 h; E3, 6 h), freezing rates (FR1, manu... more SUMMARY The effects of equilibration times (E1, 2 h; E2, 4 h; E3, 6 h), freezing rates (FR1, manual, 5 cm above liquid nitrogen (LN 2) for 10 min, plunging in LN 2 ; FR2, programmable ultra-fast, holding at +4 °C for 2 min, from 4 to À10 °C at À10 °C/min, from À10 to À20 °C at À15 °C/min, from À20 to À120 °C at À60 °C/min, holding at À120 °C for 30 sec, plunging in LN 2), and thawing rates (T1, 37 °C for 30 sec; T2, 50 °C for 15 sec; T3, 70 °C for 7 sec) were evaluated on quality of buffalo bull spermatozoa. Progressive motility (%), rapid velocity (%), average path velocity (VAP, lm/s), straight line velocity (VSL, lm/s), and mitochondrial transmembrane potential (%) were higher (p &lt; 0.05) with E2, FR2, and T3 compared to other groups. Sperm curved line velocity (VCL, lm/s) was higher (p &lt; 0.05) with E2 and FR2 compared to other groups. Sperm straightness (%) and linearity (LIN, %) were higher (p &lt; 0.05) with E2 compared to other groups. Sperm LIN was affected (p &lt; 0.05) with T3 compared to other groups. Supravital-plasma membrane integrity (%), viability and acrosome integrity (%) of spermatozoa were higher (p &lt; 0.05) with E2 and FR2 compared to other groups. Sperm DNA integrity (%) was higher (p &lt; 0.05) with FR2 and T1 compared to other groups. We concluded that inclusion of 4 h-equilibration time, programmable ultra-fast freezing rate, and rapid thawing at 70 °C for 7 sec in cryopreservation protocol improves the post-thaw quality of buffalo bull spermatozoa.
Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermato... more Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermatozoa. Semen from each of the five bulls (n = 3 replicates, six ejaculates/bull, a total of 30 ejaculates) was diluted in Tris-citric acid extender containing curcumin (0.5, 1.0, 1.5 or 2.0 mM) or control. At pre-freezing and post-thawing, total antioxidant contents (μM/L) and lipid peroxidation levels (μM/ml) were higher (p &lt; .05) and lower (p &lt; .05) respectively, with 1.5 and 2.0 mM compared to 0.5 and 1.0 mM curcumin and control. At post-thawing, progressive motility (PM, %) and rapid velocity (RV, %) were higher (p &lt; .05) with 1.5 mM compared to other doses of curcumin and control (except in case of RV, 1.5 was similar with 1.0 mM). Kinematics (average path velocity, μm/s; straight-line velocity, μm/s; curved-line velocity, μm/s; straightness, %; linearity, %), in vitro longevity (%, PM and RV) and DNA integrity (%) at post-thawing were higher (p &lt; .05) with 1.5 mM compared to control. At post-thawing, supravital plasma membrane integrity (%) and viable spermatozoa with intact acrosome (%) were higher with 1.5 compared to 2.0 mM curcumin and control. We concluded that freezability of water buffalo spermatozoa is improved with the addition of 1.5 mM curcumin in extender.
This study was primarily designed to evaluate the effect of different concentrations of ultraviol... more This study was primarily designed to evaluate the effect of different concentrations of ultraviolet (UV)-C-irradiated chicken egg yolk plasma (EYP; v:v; 10%, P1; 15%, P2; 20%, P3) or 20% (v:v) of whole chicken egg yolk (WCEY) in tris-citric acid (TCA) extender on water buffalo sperm quality during cryopreservation (postdilution, PD; postequilibration, PE; post-thawing, PT). Also the effect of best evolved concentration of UV-C-irradiated EYP in extender on in vivo fertility of buffalo spermatozoa was evaluated. At PE and PT, computer-assisted sperm analysis progressive motility (PM, %) was significantly higher in P3 compared with P1 and WCEY. Rapid velocity (RV, %) was higher (P < 0.05) in P3 compared with P1 and WCEY during cryopreservation (PD, PE, and PT). Average path velocity (μm/s) and straight line velocity (μm/s) were higher (P < 0.05) in P2 and P3 than WCEY at PE and PT. The decline percentage (%, longevity) in PM and RV was lower (P < 0.05) in P3 compared with WCE...
Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermato... more Effects of curcumin as antioxidant in extender were evaluated on freezability of buffalo spermatozoa. Semen from each of the five bulls (n = 3 replicates, six ejaculates/bull, a total of 30 ejaculates) was diluted in Tris-citric acid extender containing curcumin (0.5, 1.0, 1.5 or 2.0 mM) or control. At pre-freezing and post-thawing, total antioxidant contents (μM/L) and lipid peroxidation levels (μM/ml) were higher (p < .05) and lower (p < .05) respectively, with 1.5 and 2.0 mM compared to 0.5 and 1.0 mM curcumin and control. At post-thawing, progressive motility (PM, %) and rapid velocity (RV, %) were higher (p < .05) with 1.5 mM compared to other doses of curcumin and control (except in case of RV, 1.5 was similar with 1.0 mM). Kinematics (average path velocity, μm/s; straight-line velocity, μm/s; curved-line velocity, μm/s; straightness, %; linearity, %), in vitro longevity (%, PM and RV) and DNA integrity (%) at post-thawing were higher (p < .05) with 1.5 mM compared...
This study was primarily designed to evaluate the effect of different concentrations of ultraviol... more This study was primarily designed to evaluate the effect of different concentrations of ultraviolet (UV)-C-irradiated chicken egg yolk plasma (EYP; v:v; 10%, P1; 15%, P2; 20%, P3) or 20% (v:v) of whole chicken egg yolk (WCEY) in tris-citric acid (TCA) extender on water buffalo sperm quality during cryopreservation (postdilution, PD; postequilibration, PE; post-thawing, PT). Also the effect of best evolved concentration of UV-C-irradiated EYP in extender on in vivo fertility of buffalo spermatozoa was evaluated. At PE and PT, computer-assisted sperm analysis progressive motility (PM, %) was significantly higher in P3 compared with P1 and WCEY. Rapid velocity (RV, %) was higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P3 compared with P1 and WCEY during cryopreservation (PD, PE, and PT). Average path velocity (μm/s) and straight line velocity (μm/s) were higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P2 and P3 than WCEY at PE and PT. The decline percentage (%, longevity) in PM and RV was lower (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P3 compared with WCEY during 2 hours incubation under in vitro condition at PT. Supravital plasma membrane integrity (%) was higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P2 and P3 compared with control at different stages (PE and PT). Mitochondrial transmembrane potential (%) was higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P2 and P3 compared with P1 and WCEY at different stages (PD and PT). Percentage of viable sperm with intact acrosome, and sperm DNA integrity (%) were higher (P &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05) in P2 and P3 compared with WCEY at PT. The in vivo fertility rate (%) was significantly higher with P3 compared with WCEY (76.61 vs. 64.49). In conclusion, WCEY (20%) can be replaced with UV-C-irradiated chicken EYP (20%) in TCA extender for cryopreservation of water buffalo spermatozoa.
The receptors for sulphonylurea (SURs) are known to be expressed in the mouse kidney, but their e... more The receptors for sulphonylurea (SURs) are known to be expressed in the mouse kidney, but their expression in the ureter is undefined. Owing to the physiological and pharmacological significance of SURs, the localization of SUR in ureters of adult mice and rats was investigated through immunohistochemistry. Animals were perfused transcardially with 4% paraformaldehyde and tissues were processed for immunohistochemistry using polyclonal antisera against SUR2A and SUR2B (SUR1) receptor proteins. Sections were incubated with primary and secondary antisera and developed with aminoethylcarbazole as a chromogen. A differentiated localized staining pattern of SUR proteins in rat and mouse ureters is demonstrated. In the mouse, immunoreactivity of SUR2A was predominantly confined to the cytoplasmic portion of epithelial cells and blood vessels, with comparatively low-level staining found in smooth muscle. In contrast, SUR2B (SUR1) immunoreactivity was absent in mouse ureters. In rats, SUR2A...
Pfl�gers Archiv European Journal of Physiology, 1999
The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glib... more The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glibenclamide, which are widely used as oral hypoglycaemic agents. Sulphonylureas have also been shown to affect urine flow and salt excretion by the kidney; therefore, the use of these drugs may have important implications for the pharmacological manipulation of renal salt handling. The purpose of the present investigation was to increase our understanding of the possible role of SUR in the regulation of renal function by determining the distribution of SUR isoforms within mouse kidney. Immunostaining with anti-SUR antisera revealed specific staining of SUR2B in distal nephron segments of mouse kidney. A diffuse, low level staining was observed in proximal tubules in the inner cortical region. No evidence was found for the presence of SUR2B in intra-renal blood vessels. Reverse-transcription polymerase chain reaction and Western blotting experiments indicated that SUR2B is the only known isoform expressed. These data demonstrate that SUR2B in mouse kidney is expressed in tubule regions that are critical in determining renal salt excretion.
The receptors for sulphonylurea (SURs) are known to be expressed in the mouse kidney, but their e... more The receptors for sulphonylurea (SURs) are known to be expressed in the mouse kidney, but their expression in the ureter is undefined. Owing to the physiological and pharmacological significance of SURs, the localization of SUR in ureters of adult mice and rats was investigated through immunohistochemistry. Animals were perfused transcardially with 4% paraformaldehyde and tissues were processed for immunohistochemistry using polyclonal antisera against SUR2A and SUR2B (SUR1) receptor proteins. Sections were incubated with primary and secondary antisera and developed with aminoethylcarbazole as a chromogen. A differentiated localized staining pattern of SUR proteins in rat and mouse ureters is demonstrated. In the mouse, immunoreactivity of SUR2A was predominantly confined to the cytoplasmic portion of epithelial cells and blood vessels, with comparatively low-level staining found in smooth muscle. In contrast, SUR2B (SUR1) immunoreactivity was absent in mouse ureters. In rats, SUR2A immunoreactivity was localized only in the blood vessels, while SUR2B (SUR1) immunoreactivity was localized in the epithelial cell cytoplasm. Tissue specificity of SUR is demonstrated in the two species of rodents and suggests a role of SUR proteins in urinary metabolism pertaining possibly to salt handling and maintenance of the smooth muscle tone.
Pflugers Archiv-European Journal of Physiology, 1999
The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glib... more The sulphonylurea receptor (SUR) is the site of action for sulphonylurea derivatives such as glibenclamide, which are widely used as oral hypoglycaemic agents. Sulphonylureas have also been shown to affect urine flow and salt excretion by the kidney; therefore, the use of these drugs may have important implications for the pharmacological manipulation of renal salt handling. The purpose of the present investigation was to increase our understanding of the possible role of SUR in the regulation of renal function by determining the distribution of SUR isoforms within mouse kidney. Immunostaining with anti-SUR antisera revealed specific staining of SUR2B in distal nephron segments of mouse kidney. A diffuse, low level staining was observed in proximal tubules in the inner cortical region. No evidence was found for the presence of SUR2B in intra-renal blood vessels. Reverse-transcription polymerase chain reaction and Western blotting experiments indicated that SUR2B is the only known isoform expressed. These data demonstrate that SUR2B in mouse kidney is expressed in tubule regions that are critical in determining renal salt excretion.
The objective of the study was to devise a cryoprotection synergism between glycerol and dimethyl... more The objective of the study was to devise a cryoprotection synergism between glycerol and dimethyl sulfoxide (DMSO) for water buffalo sper-matozoa. Additionally, the effect of best evolved concentrations of glycerol and DMSO in extender was assessed on in vivo fertility of buffalo spermatozoa. Ejaculates (n = 30) were equally distributed into five aliquots; first aliquot was diluted at 37 °C in extender having 7 % glycerol (control); second aliquot was diluted at 37 °C as well as at 4 °C in extender having 3.5 % DMSO (Group 1); third aliquot was diluted at 37 °C in extender having 3.5 % glycerol and then at 4 °C in extender having 3.5 % DMSO (Group 2); fourth aliquot was diluted at 37 °C in extender having 3.5 % DMSO and then at 4 °C in extender having 3.5 % glycerol (Group 3); fifth aliquot was diluted in extenders having 1.75 % glycerol and 1.75 % DMSO at 37 as well as at 4 °C (Group 4). At post thawing, sperm progressive motility (%), rapid velocity (%), average path velocity (lm/s), curved line velocity (lm/s), in vitro longevity (%), structural and functional integrity of plasmalemma (%), mitochondrial transmembrane potential (%) and viable sperm with intact acrosome (%) were higher (P \ 0.05) in Group 4 compared to other treatment groups and control. Regarding sperm DNA integrity (%); it was higher (P \ 0.05) in Group 4 compared to Group 1, 3 and control. The in vivo fertility (%) of buffalo sperma-tozoa was significantly higher with Group 4 compared to control (69.45 vs. 59.81). In conclusion, synergism exists between glycerol and DMSO (Group 4) in improving the quality and in vivo fertility of cryop-reserved water buffalo spermatozoa.
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Papers by I. Z. Qureshi