Eur J Clin Invest 2010; 40 (8): 756–758Background Thyroid‐stimulating hormone (TSH) measurement ... more Eur J Clin Invest 2010; 40 (8): 756–758Background Thyroid‐stimulating hormone (TSH) measurement plays a major role in the diagnosis of thyroid disorders. Despite the good quality of immunochemical tests measuring TSH levels, the presence of interfering substances can sometimes alter the TSH results.Design We reported the case of a 79‐year‐old man affected by primary autoimmune hypothyroidism hospitalized for pneumonia. A TSH value > 100 mIU L‐1 (reference: 0.44 mIU L‐1) was found at admission. No signs and symptoms of hypothyroidism were found upon clinical examination and serum concentration of the free thyroxine (FT4) was normal.Results Serum treatment in heterophile antibody blocking tubes did not change the TSH result in our assay, while normal levels were found in a different immunoassay method. An abnormal pattern was found in protein electrophoresis at admission, with IgG / j and IgM / k monoclonal bands proved in immunofixation. Interestingly, the disappearance of mono...
International Journal of Peptide and Protein Research, 2009
1. In mesophilic cells (37°C culture) of B. stearothermophilus a high molecular weight aminopepti... more 1. In mesophilic cells (37°C culture) of B. stearothermophilus a high molecular weight aminopeptidase (APIm) was found in the “membrane” fraction after lysozyme treatment and ultrasonication. APIm, which is also present in 55°C cells but in a 30-fold lower amount, could be extracted from the “membranes” by carbonate-bicarbonate buffer pH 9.6 and was further purified. APIm has a mol. wt. of about 400,000 and has an oligomeric structure (mol. wt. of subunits 46,000 ± 3,000). The “membrane-bound” and the soluble enzyme differ in thermostability: The former bound-enzyme is more thermostable. Specificity: Dipeptides are poor and tri- and tetrapeptides are good substrates with Gly, Leu, Ala or Met at the N-terminus. In tripeptides the enzyme splits preferentially the first (N-terminal) peptide bond. 2. Aminopeptidase III (APIII), which is synthesized in 37°C cultures of B. stearothermophilus in a 10-fold higher amount than in 55°C cultures, was purified to homogeneity by a four-step purification procedure. Mol. wt. of APIII: 108,000 and of the subunits 47,500 ± 3,000 (dimer). APIII is not thermostable at 60–70°C. The amino acid composition of APIII for some amino acids (Lys, His, Thr, Pro, Ala, Met, Ile, Tyr, Phe) is very similar to the composition of APII, and for Lys, His, Arg, Thr, Glu, Pro, Phe it is similar to that for thermophilic API. The most striking feature of this enzyme is its specificity as aminotripeptidase.
Proceedings of the National Academy of Sciences, 1980
Iodination of toxin II from the sea anemone Anemonia sulcata gives a labeled monoiododerivative t... more Iodination of toxin II from the sea anemone Anemonia sulcata gives a labeled monoiododerivative that retains 80% of the original neurotoxicity. This derivative binds specifically to rat brain synaptosomes at 20 degrees C and pH 7.4 with a second-order rate constant of association ka = 4.6 x 10(4) M-1 sec-1 and a first-order rate constant of dissociation kd = 1.1 x 10(-2) sec-1. The binding occurs on the Na+ channel at a binding site distinct from that of other gating system toxins like batrachotoxin, veratridine, grayanotoxin, aconitine, and pyrethroids. The maximal binding capacity Bmax is 3.2 pmol/mg of protein (i.e., about two sea anemone toxin binding sites per tetrodotoxin binding site) and the Kd is 240 nM for the monoiododerivative and 150 nM for the native toxin. Corresponding binding parameters for the association of a 125I-labeled derivative of toxin II from the scorpion Androctonus australis Hector are Bmax = 0.3 pmol/mg of protein and Kd = 1 nM, whereas the Kd of the unm...
The purification of axonal membranes of crustaceans was followed by measuring enrichment in [3H]t... more The purification of axonal membranes of crustaceans was followed by measuring enrichment in [3H]tetrodotoxin binding capacity and in Na+, K+-ATPase activity. A characteristic of these membranes is their high content of lipids and their low content of protein as compared to other types of plasmatic membranes. The axonal membrane contains myosin-like, actin-like, tropomyosin-like, and tubulin-like proteins. It also contains Na+, K+-ATPase and acetylcholinesterase. The molecular weights of these two enzymes after solubilization are 280,000 and 270,000, respectively. The molecular weights of the catalytic subunits are 96,000 for ATPase and 71,000 for acetylcholinesterase. We confirmed the presence of a nicotine binding component in the axonal membrane of the lobster but we have been unable to find [3H]nicotine binding to crab axonal membranes. The binding to axonal membranes og of the sodium channel, has been studied in detail. The dissociation constant for the binding of [3H]tetrodotoxin to the axonal membrane receptor is 2.9 nM at pH 7.4. The concentration of the tetrodotoxin receptor in crustacean membranes is about 10 pmol/mg of membrane protein, 7 times less than the acetylcholinesterase, 30 times less than the Na+, K+-ATPase, and 30 times less than the nicotine binding component in the lobster membrane. A reasonable estimate indicates that approximately only one peptide chain in 1000 constitutes the tetrodotoxin binding part of the sodium channel in the axonal membrane. Veratridine, which acts selectively on the resting sodium permeability, binds to the phospholipid part of the axonal membrane. [3H]Veratridine binding to membranes parallels the electrophysiological effect. Veratridine and tetrodotoxin have different receptor sites. Although tetrodotoxin can repolarize the excitable membrane of a giant axon depolarized by veratridine, veratridine does not affect the binding of [3H]tetrodotoxin to purified axonal membranes. Similarly, tetrodotoxin does not affect the binding of [3H]veratridine to axonal membranes. Scorpion neurotoxin I, a presynaptic toxin which affects both the Na+ and the K+ channels, does not interfere with the binding of [3H]tetrodotoxin or [3H]veratridine to axonal membranes. Tetrodotoxin, veratridine, and scorpion neurotoxin I, which have in common the perturbation of the normal functioning of the sodium channel, act upon three different types of receptor sites.
Biochimica et Biophysica Acta (BBA) - Biomembranes, 1981
Freeze-fracture electron microscopy was used to follow morphological changes induced by Naja moss... more Freeze-fracture electron microscopy was used to follow morphological changes induced by Naja mossambica mossambica venom cardiotoxins on crab axonal membranes and their lipids. It was shown that the extent of morphological changes depended drastically on the purity of cardiotoxin preparations and on their nature. Highly purified cardiotoxin induced mainly fusion of membrane or lipid vesicles. The extent of fusion and other morphological changes depended on the nature of cardiotoxin used: VII4 cardiotoxin induced only fusion while VII1 led to further modifications of membranes and liposomes. The most spectacular morphological changes were observed with axonal membranes treated with cardiotoxin containing traces of venom phospholipase A2. At low cardiotoxin concentration (10-7-10-s M) important intramembrane particle aggregation was observed and at higher concentrations (more than 10-4 M) intramembrane particles disappeared from the membrane and were found in solution. The membrane vesicles, devoid of intramembrane particles, were observed to fuse rapidly into liposome-like aggregates. These morphological changes are interpreted as being due to the removal of intrinsic membrane proteins from the membrane by the combined action of cardiotoxin and phospholipase A2.
Biochimica et Biophysica Acta (BBA) - Biomembranes, 1981
The Pfitzner-Moffatt oxidation procedure has been used to prepare two new photoactivable derivati... more The Pfitzner-Moffatt oxidation procedure has been used to prepare two new photoactivable derivatives of tetrodotoxin that have been synthesized with high specific radioactivities (17.5 Ci/mmol and 30 Ci/mmol). They specifically bind to axonal membranes with affinities of 5.2-14.2 nM. They dissociate from their membrane complex with half-lives of 10.8 and 20 min. In the dark, these compounds give a reversible block of the sodium channels. After ultraviolet irradiation, they induce an irreversible blockade of the nerve channels.
To study the effect of peritoneal (PF) and follicular fluids (FF from the same woman) as well as ... more To study the effect of peritoneal (PF) and follicular fluids (FF from the same woman) as well as of given volumetric combinations thereof on sperm motility and acrosomal reactivity. Prospective. Peritoneal fluid and FF were incubated separately or in given volumetric combinations (PF/FF = 100/0, 75/25, 50/50, 25/75, 0/100; vol/vol) with swim-up sperm suspensions. In vitro fertilization and general infertility clinic and laboratories. Women participants of the gamete intrafallopian transfer program (motility study, n = 20; acrosomal reaction study, n = 14). Sperm donors of the artificial insemination program and men with given sperm parameters. Hormonal stimulation. Laparoscopy. Progressive velocity and percentage of motile gametes measured with multiple-exposure photography. Acrosomal reactivity measured by an immunofluorescent technique. Follicular fluid always influenced progressive motility and also sustained the number of motile gametes, as function of time, better than PF or the PF/FF mixtures (P less than 0.05). The acrosomal reactivity of sperm incubated in the various PF/FF combinations was low; after 5 hours only the FF-sperm suspensions showed a significant enhancement of acrosomally reacted gametes. At ovulation, FF transmit positive (motility- and acrosomal reactivity-enhancing) signals to sperm, whereas PF may transmit positive, neutral, or negative signals (noise signals). The volumetric combination of FF and PF in the tubal environment, which may differ from cycle to cycle and from woman to woman, could therefore result in synergic (or antagonistic) effects on the sperm fertility potential.
Forty-three follicular fluids (FFs) obtained during laparoscopy were tested in vitro for their ef... more Forty-three follicular fluids (FFs) obtained during laparoscopy were tested in vitro for their effect(s) on sperm motility using gametes obtained by the swim-up procedure. Both the proportion of motile sperm and the velocity distribution patterns were evaluated as function of time by multiple-exposure photography technique. At the various incubation periods considered, all FFs maintained or then enhanced sperm motility as compared with the paired control suspension incubated with a sperm survival medium. The results of the sperm contact test for FFs from women who achieved pregnancy versus FFs from women who remained infertile were not significantly different for both parameters measured. Comparing these with our previously reported results, we may hypothesize that FF released at ovulation into the peritoneal cavity may counteract some sperm-immobilizing effect of peritoneal fluid, thereby increasing the fertility potential of the male gametes. Fertil Steril 55:619, 1991 At present, follicular fluid (FF) is considered one of the fluids of the female genital tract that could be involved in sperm capacitation, hyperactivation, and acrosome reaction. 1-4 Such phenomena appear to be species-specific, 1 • 5 correlated with particular FF fractions, 3 • 5 • 6 and dose-dependent. 2 • 4 Furthermore, incorporation of native FF in sperm preparation procedures has been claimed to minimize false-negative results in the sperm penetration assay (SPA) 7 and to increase pregnancy rates (PRs) in assisted reproduction methods. 8 • 9 Concerning the effect of FF on sperm motility, there are at present a limited number of studies, and often their results are not in agreement. The scope
Seminal plasma (n = 12) from cystic fibrosis (CF) patients were analyzed by gel-electrophoresis u... more Seminal plasma (n = 12) from cystic fibrosis (CF) patients were analyzed by gel-electrophoresis using seminal plasma and expressed prostatic secretion from fertile men as controls. Heavy precipitation at the entering position of the gel and streaking in the gel matrix was observed, demonstrating a reduced solubility of seminal proteins in CF. Comparison of the protein patterns evidenced that CF-seminal plasma (CF-SP) mainly consisted of prostatic components. Although lactoferrin was undetectable in all samples, trace amounts of low molecular weight proteins were observed in two patients. This latter finding could imply that CF-SP may contain proteolytic fragments of prostatic and/or vesicular proteins or de novo synthesized components.
International Journal of Gynecology & Obstetrics, 1990
The aim of this study was to analyze in vitro the effect(s) of peritoneal fluid (PF) on sperm mot... more The aim of this study was to analyze in vitro the effect(s) of peritoneal fluid (PF) on sperm motility. Seventy PFs obtained during laparoscopy were tested on motile-rich sperm suspensions. Proportion of motile sperm and velocity distribution were evaluated by multiple-exposure photography technique. At time (t) = 0, PFs increased both sperm parameters as compared with control (P < 0.01). Maximum effect was observed at t = 5 hours: 32 (45%) PFs increasedand5 (7%) decreased the proportion of motile sperm, while 8 (11 %) PFs increased and 4 (6%) decreased sperm velocity. No correlation was found between a particular infertile group and a definite negative effect. However, 70% of PFs from fertile women maintained or increased the proportion of motile sperm at t = 5 hours, compared with 36% in the total infertile group. Comparison of the sperm motility effect(s) of a given PF on different ejaculates revealed that the effects observed also were influenced by the sperm sample tested. In conclusion, PFs can maintain or increase the motility of spermatozoa as function of time. However, some PFs can inhibit sperm motility and these effect(s) can be influenced by the sperm sample. Fertil Steril52:113, 1989
Photoaffinity labelling by 8-N3-/32P/cAMP of human sperm homogenates with normal pathology and no... more Photoaffinity labelling by 8-N3-/32P/cAMP of human sperm homogenates with normal pathology and normal progressive motility of the gametes (N) revealed the presence of 2 specific CAMP-binding activities with MW 47 000 (R-I) and 52 000 (R-11). Similar amounts of R-I and R-I1 were found in sperm homogenates obtained from semen samples with either reduced progressive motility (A50: 30-5076 progressive motility and A30: < 30% progressive motility) or abnormal morphology of the gametes (T: 5-30% normal heads). In contrast, the seminal plasma of the respective semen samples of N, A50, A30 and T incorporated the 8-azido CAMP photolabel in similar quantities into 2 additional CAMP-binding proteins with MW 42000 and 37000. Quantitative analysis of the photoaffinity labelling of the seminal plasma obtained from normal (N) or subnormal semen (A50, A30, T) showed that CAMP was predominantly bound to the 37 000 protein. Furthermore, the cAMPdependent protein kinase and CAMP-binding activities of sperm homogenates were quantitatively comparable within the 4 groups. These results provide the first evidence that neither quantitative nor qualitative differences in the molecular and/or enzymatic properties of cAMPdependent protein kinases can be detected between normal human sperm (N) and those with either reduced progressive motility (A50, A30) or abnormal morphology (T).
It was recently demonstrated that the protein kinase activity of human seminal plasma was cAMP-in... more It was recently demonstrated that the protein kinase activity of human seminal plasma was cAMP-independent (Majumder, 1978). However, seminal plasma is not an intrinsic component of this fluid, that it is not produced by the accessory male gland secretions, and that it may originate from the spermatozoa during semen liquefaction.
Objective: To determine the effects of peritoneal and follicular fluids (PFs, FFs) on sperm acros... more Objective: To determine the effects of peritoneal and follicular fluids (PFs, FFs) on sperm acrosomal reaction (AR). Design: Prospective, randomized study. Setting: Three hospital-based infertility units. Patients: Twenty-three women participating in GIFT programs; 23 men participating in AIH programs. Interventions: Hormonal stimulation after buserelin desensitation; laparoscopy 36 hrs after hCG injection. Main outcome measure: Percentage of acrosomally-reacted sperm. Results: Compared with Earle's medium (control), moderate but significant increases of ARs were observed as function 1) of the relative content of FF in the incubation medium and 2) as function of time (these increases were constantly lower than those registered for the respective positive control, i.e. 2X frozen/thawed sperm). In contrast, when PF alone was present in the incubation medium, no such effects on AR were registered. Conclusions: FF and mixtures of PF and FF-but not PF alone-seem to induce some rapid and time-dependent processes which finally lead to an AR. Therefore, and independently on the infertlility cause (tubal, male-dependent, unexplained infertlity), PF seems able to exert effects on sperm motility (Revelli et al., Fertil. Steril 57, 654-60 [1992]) while maintaining an unreacted sperm status.
Eur J Clin Invest 2010; 40 (8): 756–758Background Thyroid‐stimulating hormone (TSH) measurement ... more Eur J Clin Invest 2010; 40 (8): 756–758Background Thyroid‐stimulating hormone (TSH) measurement plays a major role in the diagnosis of thyroid disorders. Despite the good quality of immunochemical tests measuring TSH levels, the presence of interfering substances can sometimes alter the TSH results.Design We reported the case of a 79‐year‐old man affected by primary autoimmune hypothyroidism hospitalized for pneumonia. A TSH value > 100 mIU L‐1 (reference: 0.44 mIU L‐1) was found at admission. No signs and symptoms of hypothyroidism were found upon clinical examination and serum concentration of the free thyroxine (FT4) was normal.Results Serum treatment in heterophile antibody blocking tubes did not change the TSH result in our assay, while normal levels were found in a different immunoassay method. An abnormal pattern was found in protein electrophoresis at admission, with IgG / j and IgM / k monoclonal bands proved in immunofixation. Interestingly, the disappearance of mono...
International Journal of Peptide and Protein Research, 2009
1. In mesophilic cells (37°C culture) of B. stearothermophilus a high molecular weight aminopepti... more 1. In mesophilic cells (37°C culture) of B. stearothermophilus a high molecular weight aminopeptidase (APIm) was found in the “membrane” fraction after lysozyme treatment and ultrasonication. APIm, which is also present in 55°C cells but in a 30-fold lower amount, could be extracted from the “membranes” by carbonate-bicarbonate buffer pH 9.6 and was further purified. APIm has a mol. wt. of about 400,000 and has an oligomeric structure (mol. wt. of subunits 46,000 ± 3,000). The “membrane-bound” and the soluble enzyme differ in thermostability: The former bound-enzyme is more thermostable. Specificity: Dipeptides are poor and tri- and tetrapeptides are good substrates with Gly, Leu, Ala or Met at the N-terminus. In tripeptides the enzyme splits preferentially the first (N-terminal) peptide bond. 2. Aminopeptidase III (APIII), which is synthesized in 37°C cultures of B. stearothermophilus in a 10-fold higher amount than in 55°C cultures, was purified to homogeneity by a four-step purification procedure. Mol. wt. of APIII: 108,000 and of the subunits 47,500 ± 3,000 (dimer). APIII is not thermostable at 60–70°C. The amino acid composition of APIII for some amino acids (Lys, His, Thr, Pro, Ala, Met, Ile, Tyr, Phe) is very similar to the composition of APII, and for Lys, His, Arg, Thr, Glu, Pro, Phe it is similar to that for thermophilic API. The most striking feature of this enzyme is its specificity as aminotripeptidase.
Proceedings of the National Academy of Sciences, 1980
Iodination of toxin II from the sea anemone Anemonia sulcata gives a labeled monoiododerivative t... more Iodination of toxin II from the sea anemone Anemonia sulcata gives a labeled monoiododerivative that retains 80% of the original neurotoxicity. This derivative binds specifically to rat brain synaptosomes at 20 degrees C and pH 7.4 with a second-order rate constant of association ka = 4.6 x 10(4) M-1 sec-1 and a first-order rate constant of dissociation kd = 1.1 x 10(-2) sec-1. The binding occurs on the Na+ channel at a binding site distinct from that of other gating system toxins like batrachotoxin, veratridine, grayanotoxin, aconitine, and pyrethroids. The maximal binding capacity Bmax is 3.2 pmol/mg of protein (i.e., about two sea anemone toxin binding sites per tetrodotoxin binding site) and the Kd is 240 nM for the monoiododerivative and 150 nM for the native toxin. Corresponding binding parameters for the association of a 125I-labeled derivative of toxin II from the scorpion Androctonus australis Hector are Bmax = 0.3 pmol/mg of protein and Kd = 1 nM, whereas the Kd of the unm...
The purification of axonal membranes of crustaceans was followed by measuring enrichment in [3H]t... more The purification of axonal membranes of crustaceans was followed by measuring enrichment in [3H]tetrodotoxin binding capacity and in Na+, K+-ATPase activity. A characteristic of these membranes is their high content of lipids and their low content of protein as compared to other types of plasmatic membranes. The axonal membrane contains myosin-like, actin-like, tropomyosin-like, and tubulin-like proteins. It also contains Na+, K+-ATPase and acetylcholinesterase. The molecular weights of these two enzymes after solubilization are 280,000 and 270,000, respectively. The molecular weights of the catalytic subunits are 96,000 for ATPase and 71,000 for acetylcholinesterase. We confirmed the presence of a nicotine binding component in the axonal membrane of the lobster but we have been unable to find [3H]nicotine binding to crab axonal membranes. The binding to axonal membranes og of the sodium channel, has been studied in detail. The dissociation constant for the binding of [3H]tetrodotoxin to the axonal membrane receptor is 2.9 nM at pH 7.4. The concentration of the tetrodotoxin receptor in crustacean membranes is about 10 pmol/mg of membrane protein, 7 times less than the acetylcholinesterase, 30 times less than the Na+, K+-ATPase, and 30 times less than the nicotine binding component in the lobster membrane. A reasonable estimate indicates that approximately only one peptide chain in 1000 constitutes the tetrodotoxin binding part of the sodium channel in the axonal membrane. Veratridine, which acts selectively on the resting sodium permeability, binds to the phospholipid part of the axonal membrane. [3H]Veratridine binding to membranes parallels the electrophysiological effect. Veratridine and tetrodotoxin have different receptor sites. Although tetrodotoxin can repolarize the excitable membrane of a giant axon depolarized by veratridine, veratridine does not affect the binding of [3H]tetrodotoxin to purified axonal membranes. Similarly, tetrodotoxin does not affect the binding of [3H]veratridine to axonal membranes. Scorpion neurotoxin I, a presynaptic toxin which affects both the Na+ and the K+ channels, does not interfere with the binding of [3H]tetrodotoxin or [3H]veratridine to axonal membranes. Tetrodotoxin, veratridine, and scorpion neurotoxin I, which have in common the perturbation of the normal functioning of the sodium channel, act upon three different types of receptor sites.
Biochimica et Biophysica Acta (BBA) - Biomembranes, 1981
Freeze-fracture electron microscopy was used to follow morphological changes induced by Naja moss... more Freeze-fracture electron microscopy was used to follow morphological changes induced by Naja mossambica mossambica venom cardiotoxins on crab axonal membranes and their lipids. It was shown that the extent of morphological changes depended drastically on the purity of cardiotoxin preparations and on their nature. Highly purified cardiotoxin induced mainly fusion of membrane or lipid vesicles. The extent of fusion and other morphological changes depended on the nature of cardiotoxin used: VII4 cardiotoxin induced only fusion while VII1 led to further modifications of membranes and liposomes. The most spectacular morphological changes were observed with axonal membranes treated with cardiotoxin containing traces of venom phospholipase A2. At low cardiotoxin concentration (10-7-10-s M) important intramembrane particle aggregation was observed and at higher concentrations (more than 10-4 M) intramembrane particles disappeared from the membrane and were found in solution. The membrane vesicles, devoid of intramembrane particles, were observed to fuse rapidly into liposome-like aggregates. These morphological changes are interpreted as being due to the removal of intrinsic membrane proteins from the membrane by the combined action of cardiotoxin and phospholipase A2.
Biochimica et Biophysica Acta (BBA) - Biomembranes, 1981
The Pfitzner-Moffatt oxidation procedure has been used to prepare two new photoactivable derivati... more The Pfitzner-Moffatt oxidation procedure has been used to prepare two new photoactivable derivatives of tetrodotoxin that have been synthesized with high specific radioactivities (17.5 Ci/mmol and 30 Ci/mmol). They specifically bind to axonal membranes with affinities of 5.2-14.2 nM. They dissociate from their membrane complex with half-lives of 10.8 and 20 min. In the dark, these compounds give a reversible block of the sodium channels. After ultraviolet irradiation, they induce an irreversible blockade of the nerve channels.
To study the effect of peritoneal (PF) and follicular fluids (FF from the same woman) as well as ... more To study the effect of peritoneal (PF) and follicular fluids (FF from the same woman) as well as of given volumetric combinations thereof on sperm motility and acrosomal reactivity. Prospective. Peritoneal fluid and FF were incubated separately or in given volumetric combinations (PF/FF = 100/0, 75/25, 50/50, 25/75, 0/100; vol/vol) with swim-up sperm suspensions. In vitro fertilization and general infertility clinic and laboratories. Women participants of the gamete intrafallopian transfer program (motility study, n = 20; acrosomal reaction study, n = 14). Sperm donors of the artificial insemination program and men with given sperm parameters. Hormonal stimulation. Laparoscopy. Progressive velocity and percentage of motile gametes measured with multiple-exposure photography. Acrosomal reactivity measured by an immunofluorescent technique. Follicular fluid always influenced progressive motility and also sustained the number of motile gametes, as function of time, better than PF or the PF/FF mixtures (P less than 0.05). The acrosomal reactivity of sperm incubated in the various PF/FF combinations was low; after 5 hours only the FF-sperm suspensions showed a significant enhancement of acrosomally reacted gametes. At ovulation, FF transmit positive (motility- and acrosomal reactivity-enhancing) signals to sperm, whereas PF may transmit positive, neutral, or negative signals (noise signals). The volumetric combination of FF and PF in the tubal environment, which may differ from cycle to cycle and from woman to woman, could therefore result in synergic (or antagonistic) effects on the sperm fertility potential.
Forty-three follicular fluids (FFs) obtained during laparoscopy were tested in vitro for their ef... more Forty-three follicular fluids (FFs) obtained during laparoscopy were tested in vitro for their effect(s) on sperm motility using gametes obtained by the swim-up procedure. Both the proportion of motile sperm and the velocity distribution patterns were evaluated as function of time by multiple-exposure photography technique. At the various incubation periods considered, all FFs maintained or then enhanced sperm motility as compared with the paired control suspension incubated with a sperm survival medium. The results of the sperm contact test for FFs from women who achieved pregnancy versus FFs from women who remained infertile were not significantly different for both parameters measured. Comparing these with our previously reported results, we may hypothesize that FF released at ovulation into the peritoneal cavity may counteract some sperm-immobilizing effect of peritoneal fluid, thereby increasing the fertility potential of the male gametes. Fertil Steril 55:619, 1991 At present, follicular fluid (FF) is considered one of the fluids of the female genital tract that could be involved in sperm capacitation, hyperactivation, and acrosome reaction. 1-4 Such phenomena appear to be species-specific, 1 • 5 correlated with particular FF fractions, 3 • 5 • 6 and dose-dependent. 2 • 4 Furthermore, incorporation of native FF in sperm preparation procedures has been claimed to minimize false-negative results in the sperm penetration assay (SPA) 7 and to increase pregnancy rates (PRs) in assisted reproduction methods. 8 • 9 Concerning the effect of FF on sperm motility, there are at present a limited number of studies, and often their results are not in agreement. The scope
Seminal plasma (n = 12) from cystic fibrosis (CF) patients were analyzed by gel-electrophoresis u... more Seminal plasma (n = 12) from cystic fibrosis (CF) patients were analyzed by gel-electrophoresis using seminal plasma and expressed prostatic secretion from fertile men as controls. Heavy precipitation at the entering position of the gel and streaking in the gel matrix was observed, demonstrating a reduced solubility of seminal proteins in CF. Comparison of the protein patterns evidenced that CF-seminal plasma (CF-SP) mainly consisted of prostatic components. Although lactoferrin was undetectable in all samples, trace amounts of low molecular weight proteins were observed in two patients. This latter finding could imply that CF-SP may contain proteolytic fragments of prostatic and/or vesicular proteins or de novo synthesized components.
International Journal of Gynecology & Obstetrics, 1990
The aim of this study was to analyze in vitro the effect(s) of peritoneal fluid (PF) on sperm mot... more The aim of this study was to analyze in vitro the effect(s) of peritoneal fluid (PF) on sperm motility. Seventy PFs obtained during laparoscopy were tested on motile-rich sperm suspensions. Proportion of motile sperm and velocity distribution were evaluated by multiple-exposure photography technique. At time (t) = 0, PFs increased both sperm parameters as compared with control (P < 0.01). Maximum effect was observed at t = 5 hours: 32 (45%) PFs increasedand5 (7%) decreased the proportion of motile sperm, while 8 (11 %) PFs increased and 4 (6%) decreased sperm velocity. No correlation was found between a particular infertile group and a definite negative effect. However, 70% of PFs from fertile women maintained or increased the proportion of motile sperm at t = 5 hours, compared with 36% in the total infertile group. Comparison of the sperm motility effect(s) of a given PF on different ejaculates revealed that the effects observed also were influenced by the sperm sample tested. In conclusion, PFs can maintain or increase the motility of spermatozoa as function of time. However, some PFs can inhibit sperm motility and these effect(s) can be influenced by the sperm sample. Fertil Steril52:113, 1989
Photoaffinity labelling by 8-N3-/32P/cAMP of human sperm homogenates with normal pathology and no... more Photoaffinity labelling by 8-N3-/32P/cAMP of human sperm homogenates with normal pathology and normal progressive motility of the gametes (N) revealed the presence of 2 specific CAMP-binding activities with MW 47 000 (R-I) and 52 000 (R-11). Similar amounts of R-I and R-I1 were found in sperm homogenates obtained from semen samples with either reduced progressive motility (A50: 30-5076 progressive motility and A30: < 30% progressive motility) or abnormal morphology of the gametes (T: 5-30% normal heads). In contrast, the seminal plasma of the respective semen samples of N, A50, A30 and T incorporated the 8-azido CAMP photolabel in similar quantities into 2 additional CAMP-binding proteins with MW 42000 and 37000. Quantitative analysis of the photoaffinity labelling of the seminal plasma obtained from normal (N) or subnormal semen (A50, A30, T) showed that CAMP was predominantly bound to the 37 000 protein. Furthermore, the cAMPdependent protein kinase and CAMP-binding activities of sperm homogenates were quantitatively comparable within the 4 groups. These results provide the first evidence that neither quantitative nor qualitative differences in the molecular and/or enzymatic properties of cAMPdependent protein kinases can be detected between normal human sperm (N) and those with either reduced progressive motility (A50, A30) or abnormal morphology (T).
It was recently demonstrated that the protein kinase activity of human seminal plasma was cAMP-in... more It was recently demonstrated that the protein kinase activity of human seminal plasma was cAMP-independent (Majumder, 1978). However, seminal plasma is not an intrinsic component of this fluid, that it is not produced by the accessory male gland secretions, and that it may originate from the spermatozoa during semen liquefaction.
Objective: To determine the effects of peritoneal and follicular fluids (PFs, FFs) on sperm acros... more Objective: To determine the effects of peritoneal and follicular fluids (PFs, FFs) on sperm acrosomal reaction (AR). Design: Prospective, randomized study. Setting: Three hospital-based infertility units. Patients: Twenty-three women participating in GIFT programs; 23 men participating in AIH programs. Interventions: Hormonal stimulation after buserelin desensitation; laparoscopy 36 hrs after hCG injection. Main outcome measure: Percentage of acrosomally-reacted sperm. Results: Compared with Earle's medium (control), moderate but significant increases of ARs were observed as function 1) of the relative content of FF in the incubation medium and 2) as function of time (these increases were constantly lower than those registered for the respective positive control, i.e. 2X frozen/thawed sperm). In contrast, when PF alone was present in the incubation medium, no such effects on AR were registered. Conclusions: FF and mixtures of PF and FF-but not PF alone-seem to induce some rapid and time-dependent processes which finally lead to an AR. Therefore, and independently on the infertlility cause (tubal, male-dependent, unexplained infertlity), PF seems able to exert effects on sperm motility (Revelli et al., Fertil. Steril 57, 654-60 [1992]) while maintaining an unreacted sperm status.
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Papers by Marco Balerna