The more pieces, the better the puzzle: sperm concentration increases gametic compatibility.

The genetic benefits individuals receive from mate choice have been the focus of numerous studies, with several showing support for both intrinsic genetic benefits and compatibility effects on fertilization success and offspring viability. However, the robustness of these effects have rarely been tested across an ecologically relevant environmental gradient. In particular, sperm environment is a crucial factor determining fertilization success in many species, especially those with external fertilization. Here, we test the importance of sperm environment in mediating compatibility-based selection on fertilization using a factorial breeding design. We detected a significant intrinsic male effect on fertilization success at only one of four sperm concentrations. Compatibility effects were significant at the two highest sperm concentrations and, interestingly, the magnitude of the compatibility effect consistently increased with sperm concentration. This suggests that females are able to modify the probability of sperm-egg fusion as the amount of sperm available increases.

Novel beta-D-galactofuranose-containing high-mannose type oligosaccharides in ascorbate oxidase from Acremonium sp. HI-25.

Ascorbate oxidase from the fungus Acremonium sp. HI-25 is a copper-containing glycoprotein that catalyzes the oxidation of ascorbic acid to dehydroascorbic acid. Monosaccharide composition analysis showed that the enzyme contains exclusively N-linked oligosaccharide chains. Following liberation by hydrazinolysis/re-N-acetylation, and fractionation by HPLC on anion exchange. Amide-80 and/or octadecyl silica columns after derivatization with p-aminobenzoic ethyl ester, the structures of the twelve major neutral oligosaccharides were identified by FAB-MS, 400 MHz 1H-NMR, methylation analysis, mild acid hydrolysis, and/or sequential exoglycosidase digestions. Acremonium sp. ascorbate oxidase was found to consist of high-mannose type oligosaccharides (76.3%) having 4 to 9 mannose residues and a series of novel D-galactofuranose-containing high-mannose type oligosaccharides (18.6%) with the following structure.

Application of N-terminally truncated DNA polymerase from Thermus thermophilus (delta Tth polymerase) to DNA sequencing and polymerase chain reactions: comparative study of delta Tth and wild-type Tth polymerases.

N-Terminally truncated DNA polymerase from Thermus thermophilus (delta Tth polymerase) lacking 5'-3' exonuclease activity was used for DNA sequencing and polymerase chain reaction (PCR). In contrast to the high background of the sequencing ladder observed with the wild-type Tth polymerase, delta Tth polymerase gave readable sequencing patterns which extend up to more than 500 bases from the primer site on cycle sequencing and automated sequencing. The delta Tth polymerase was used for the standard and mutagenic PCR, and net amplification of the DNA and the mutations accumulated during PCR were analyzed. Under mutagenic PCR, the mutation rates were 7.0 x 10(-4) (Tth) and 8.3 x 10(-4) (delta Tth) per nucleotide per cycle of amplification, which were 4-9 times higher than the rates under standard PCR.

Nucleotide sequence of the uricase gene from Bacillus sp. TB-90.

The nucleotide sequence of the uricase gene from the thermophilic bacterium Bacillus sp. TB-90 was determined. The primary structure of the uricase deduced from the nucleotide sequence comprised 332 amino acids, with a total molecular mass of 37,994 Da. The molecular mass of the subunit of the uricase produced by the transformant of Escherichia coli agreed well with this value. However, the molecular mass of a subunit of the uricase produced by Bacillus sp. TB-90 was found to be 34,000 Da by SDS-PAGE. The difference between these molecular masses was attributed to processing of the C-terminal 13 amino acid residue in Bacillus sp. TB-90. Comparison of the enzymatic properties of both uricases showed that the thermostability of the uricase produced by the transformant was enhanced by about 10 degrees C in comparison to that produced by Bacillus sp. TB-90.

Cloning of the creatinine amidohydrolase gene from Pseudomonas sp. PS-7.

The gene encoding creatinine amidohydrolase (EC 3.5.2.10) was isolated from Pseudomonas sp. PS-7. The primary structure of creatinine amidohydrolase deduced from the nucleotide sequence showed that the protein is composed of 259 amino acids and has a molecular weight of 28,437. The enzymatic property of creatinine amidohydrolase produced by recombinant E. coli was identical with those by Pseudomonas sp. PS-7.

Genesis of the Austin Flint murmur: relation to mitral inflow and aortic regurgitant flow dynamics.

OBJECTIVES: This study was designed to elucidate the genesis of the Austin Flint murmur. BACKGROUND: The Austin Flint murmur is an apical diastolic rumble associated with significant aortic regurgitation. The precise mechanism of the murmur remains unclear. METHODS: The relation between the Austin Flint murmur and mitral inflow and aortic regurgitant flow dynamics was evaluated nonivasively in 13 patients with moderate to severe aortic regurgitation and 15 control subjects using phonocardiographic and pulsed and color-coded Doppler echocardiographic techniques. The severity of aortic regurgitation was determined by color-coded Doppler echocardiography on the basis of the maximal distance of the regurgitant signal. RESULTS: The direction of aortic regurgitant flow was unrelated to the presence of the Austin Flint murmur. The severity of aortic regurgitation was greater in patients with than in those without this murmur. The peak mitral inflow velocity during early diastole (E) was significantly increased, and both peak mitral inflow velocity at atrial contraction (A) and the A/E ratio were significantly decreased in patients with the Austin Flint murmur compared with values in those without this murmur or in control subjects. However, the maximal amplitude of the Austin Flint murmur did not coincide temporally with the peak mitral inflow velocity. The murmur continued both after rapid mitral inflow had ended and during diastolic mitral regurgitation. CONCLUSIONS: The increased velocity of early diastolic mitral inflow in patients with the Austin Flint murmur is due to aortic regurgitation, but rapid mitral inflow is not an essential requirement for production of the murmur. In some cases, the Austin Flint murmur may be generated by aortic regurgitant flow alone.

Determination of alpha-amylase using a new blocked substrate (3-ketobutylidene beta-2-chloro-4-nitrophenyl-maltopentaoside).

A new substrate, 3-ketobutylidene beta-2-chloro-4-nitrophenylmaltopentaoside (3KB-CNPG5), was used for the determination of alpha-amylase (EC 3.2.1.1) in serum and urine. Under this alpha-amylase assay condition, 3KB-CNPG5 is resistant to glucoamylase and alpha-glucosidase, which are auxiliary enzymes, because the 4- and 6-positions of the non-reducing-end glucose residue are modified by the 3-ketobutylidene group. The assay using 3KB-CNPG5 for alpha-amylase activity is a highly sensitive method that uses 2-chloro-4-nitrophenol (CNP) as an aglycone, and is a stable method for determination of alpha-amylase activity in biological fluids.

[Evaluation of pulmonary venous flow pattern in hypertrophied and dilated hearts: a study with transesophageal pulsed Doppler echocardiography].

In order to evaluate the clinical significance of pulmonary venous flow (PVF) pattern, transesophageal pulsed Doppler echocardiography (TEE) was performed in 25 patients with hypertrophied heart (all with hypertrophic cardiomyopathy), 15 patients with dilated heart (10 with old myocardial infarction and 5 with dilated cardiomyopathy) and 10 normal controls. Parameters obtained from the PVF pattern were compared with those of transmitral flow (MVF) pattern, % fractional shortening (%FS) of left ventricle (LV) and amplitude of mitral anular motion (MAM) during a cardiac cycle. Results were as follows: 1. PVF pattern in cases of sinus rhythm was divided into four components, atrial systolic backward flow (PVA), ventricular systolic (PVS1, PVS2) and diastolic (PVD) forward flows. 2. In patients with dilated heart, peak velocities of PVS1 and PVS2 were markedly decreased compared with those of hypertrophied and normal hearts. 3. Peak velocity of PVD in hypertrophied and dilated hearts was significantly decreased compared with that of normal controls, and PV-D/S (ratio of peak velocity of PVD to PVS2) was significantly lower in hypertrophied heart than in normal controls. 4. Time interval from the first heart sound to the peak of PVS2 (TS) was significantly longer in dilated heart, and time interval from the second heart sound to the peak of PVD (TD) was longer in hypertrophied heart than in the other two groups. 5. MAM and %FS of dilated heart were significantly lower than those in normal and hypertrophied hearts, and peak velocity of PVS2 in dilated heart group correlated well with MAM or %FS. 6. There were significant correlations among the diastolic parameters from PVD of PVF (peak velocity of PVD, PV-D/S) and early diastolic wave (D) of MVF (peak velocity, deceleration time and deceleration of rapid filling). 7. In a case of hypertrophic cardiomyopathy with mid-diastolic wave of MVF, distinct forward wave was observed after PVD of PVF, and this wave coincided in timing with the mid-diastolic wave of MVF. 8. In a case of extensive myocardial infarction with the development of severe LV dysfunction and with "normalized" pattern of MVF, peak velocities of PVD and PVA were markedly increased, and that of PVS2 was decreased. However, the peak velocities of PVD and PVA were decreased, and that of PVS2 was increased with the fair improvement of LV dysfunction and with the compensatory augmentation of atrial contraction wave (A) of MVF.(ABSTRACT TRUNCATED AT 400 WORDS)

[On the mechanisms of mitral regurgitation in rheumatic mitral valve disease: with special reference to the role of mitral valve prolapse].

To assess the role of mitral valve prolapse (MVP) in the pathogenesis of mitral regurgitation (MR) in rheumatic mitral valve disease (RMD), we performed phonocardiography (PCG), transthoracic and transesophageal two-dimensional and color Doppler (CD) echocardiography in 22 patients with RMD including three with pure mitral stenosis (MS), 11 with predominant MS, six with predominant MR and two with pure MR. Results were as follows: 1. Prolapse of the mitral valve (MV) was differentiated from systolic ballooning of the whole MV by the findings that the anterior leaflet's tip (rough zone) protruded into the left atrium with an acute angle between the body (clear zone) and rough zones of the anterior MV and that the posterior leaflet protruded markedly above the level of the mitral ring. 2. MR was detected in six patients (slight MR) by only the CD method and in 13 (mild, moderate or greater MR) by both the PCG and CD methods. 3. MR was absent or slight in five patients (three of pure MS and two of predominant MS) without valve thickening and with systolic ballooning of the whole valve due to commissural fusion. 4. Mitral valve abnormalities related to significant (mild, moderate or severe) MR were dependent on valve thickening (five patients), prolapse of the leaflet's tip toward the left atrium (four), or both (four). 5. An apical systolic click was found in only one of the nine patients with systolic ballooning, but in four of 11 with MVP. 6. The MR murmur in six of the nine patients with valve thickening showed the decrescendo or flat contour, but that in four of the eight patients with MVP showed a crescendo contour. From these results, we concluded that mitral valve prolapse should be considered as one of the important causes of mitral regurgitation in rheumatic mitral valve disease.

[Genesis and clinical significance of an apical diastolic rumble in patients with mitral Björk-Shiley valve: a Doppler echocardiographic study].

The genesis and clinical significance of an apical diastolic rumble after replacement with mitral Björk-Shiley prosthesis were studied in 20 patients by left ventricular (LV) inflow dynamics using Doppler echocardiography. 1. The rumble was audible in 17 of the 20 patients (85%). In all but one patient, its intensity was less than Levine 2/6. Duration of the rumble ranged from 248 to 544 (mean 377) msec. There was no case of prosthetic valve dysfunction. Duration of the slow filling wave (SF interval) of the apexcardiogram in 19 cases ranged from 150 to 541 (mean 317) msec, and it correlated significantly with the duration of the rumble (r = 0.95) in 15 cases. 2. In 20 patients, peak velocity of the LV inflow obtained from continuous wave Doppler echocardiograms ranged from 1.17 to 1.89 (mean 1.56) m/sec, and pressure half time (PHT) ranged from 82 to 233 (mean 134) msec. In 19 patients, the SF interval correlated significantly with PHT (r = 0.87), but not with peak inflow velocity. 3. Peak inflow velocity was significantly slower in cases without a rumble than in cases with a rumble. PHT and the SF interval tended to be shorter in the former than in the latter. 4. There was no significant difference in the SF interval, peak inflow velocity and PHT between eight patients with smaller valve sizes (less than or equal to 25 mm) and 12 with larger valve sizes (greater than or equal to 27 mm). 5. There was no obvious relation between the direction of the main LV inflow jet as assessed by Doppler color flow mapping and the presence of the rumble in 19 patients. 6. In 19 patients, the SF interval correlated significantly with left atrial dimension (r = 0.47), but it did not correlate with LV end-diastolic dimension, fractional shortening of the LV, and the peak lengthening rate of LV internal dimension. 7. Three patients with both the rumble and SF lasting 500 msec had prolonged PHT longer than 190 msec. One of these three had congestive heart failure and suspected prosthetic mitral valve stenosis. From these results, we concluded that an apical diastolic rumble after replacement with mitral Björk-Shiley valve is mainly due to the increase in resistance and velocity of the LV inflow, and that both durations of the rumble and the SF intervals as well as PHT of the Doppler echocardiogram are useful in detecting stenosis of prosthetic mitral valves.

[Relationship between regurgitant flow dynamics and cardiac physical in tricuspid regurgitation: a phono-, mechano- and Doppler echocardiographic study].

To evaluate the relationships between regurgitant flow dynamics of tricuspid regurgitation (TR) and cardiac physical signs, and to clarify the role of atrial function on central venous flow, we investigated physical signs by cardiac auscultation and palpation of the liver. In addition, phonocardiography, jugular venous and hepatic pulse tracings and Doppler echocardiographic recordings were performed. The subjects, 109 patients with Doppler-detected TR, were categorized as an SR group of 42 with sinus rhythm, an Af group of 63 with atrial fibrillation and four with sinus arrest. Thirty-five patients underwent open heart surgery before six months or more. Results were as follows: 1. In the Af group, the maximum systolic flow velocity data in the superior vena cava (SVC) and hepatic vein (HV) correlated well with the maximum tricuspid regurgitant signal area on the color Doppler echocardiogram, and systolic backward flow from the heart was more evident in the HV than in the SVC. In the SR group, however, no correlation was observed between the maximum systolic flow velocity and the TR signal area, and systolic backward flow was not evident even in cases with severe TR. 2. After open heart surgery, systolic flow velocities in the SVC and HV were significantly decreased in the SR group compared to the Af group. 3. There was close correlation between the presence of hepatomegaly and systolic backward flow towards the liver. Hepatomegaly was more marked in the Af group than in the SR group. 4. Jugular venous and hepatic pulse data correlated well with the flow velocity data in the SVC and HV and with the TR signal area. 5. Intensity of the tricuspid regurgitant murmur as estimated by the Levine's classification correlated relatively well with the systolic pressure gradient between the right ventricle and right atrium as calculated by the modified Bernoulli equation, but did not correlate with the TR signal area. From these results, we conclude that the intensity of the tricuspid regurgitant murmur and the jugular venous or hepatic pulse patterns are useful for evaluating tricuspid regurgitant dynamics, when they are applied clinically with precise recognition of their significance and limitations, and that sinus rhythm or atrial fibrillation is also an important factor.

[Assessment of prolapsing pattern of the anterior mitral valve in mitral valve prolapse: new echocardiographic diagnostic criteria].

In the present study, the ballooning pattern of the anterior mitral valve (AMV) in mitral valve prolapse (MVP) was investigated, and new diagnostic criteria for MVP were established using two-dimensional and Doppler echocardiography. The study population consisted of 164 patients with prolapse of the AMV alone, including 86 patients with idiopathic MVP, 52 associated with atrial septal defect (ASD), 17 having chordal rupture and nine associated with connective tissue disorders. There were 60 normal controls. The results were as follows: 1. The AMV was divided into two zones, clear and rough (CZ and RZ), according to the point of insertion of the strut chordae based on two-dimensional long-axis echocardiograms of the left ventricle. The severity of AMV prolapse was determined by an angle between the posterior aortic wall (PAO)-CZ and the CZ-RZ. a) Type A: Apparently there was a transitional point between CZ and RZ, despite normal PAO-CZ and CZ-RZ angles. The RZ showed mild ballooning or prolapse into the left atrium. b) Type B: Although the PAO-CZ angle was normal, the CZ-RZ angle was markedly narrowed. Therefore, prolapse of the RZ was more severe compared with that of type A. c) Type C: An overall zone of the AMV showed ballooning or prolapse into the left atrium due to a narrowed PAO-CZ angle. 2. Type B prolapse was frequently observed in idiopathic MVP (58%), the ASD group (71%) and the chordal rupture group (71%), and type C prolapse in MVP of connective tissue disorders (89%). 3. All of 18 patients (100%) with type A, 48 of 99 (48%) with type B, and 10 of 47 (21%) with type C could not be diagnosed as MVP using Gilbert's criterion. 4. Doppler mitral regurgitation (MR) was detected in 40 of the 47 patients (85%) with type C in 56 of the 99 (59%) with type B, and in seven of the 18 (39%) with type A. These results suggested that classification of the two-dimensional echocardiography of the AMV into two zones, clear and rough (CZ and RZ), could contribute to determining not only the severity of AMV prolapse, but also to the extent of myxomatous changes of the AMV and to evaluating the correlations between the degree of MVP and the incidence of MR.

[Two-dimensional and Doppler echocardiographic studies of the genesis of a high-pitched diastolic rumble in mitral stenosis: contribution of the deformed mitral orifice and eccentric mitral inflow jet].

To clarify the genesis of a high-pitched diastolic rumble in mitral stenosis, 51 patients with mitral stenosis were studied. They were subdivided into two groups based on the pitch of a rumble; six patients with a high-pitched rumble and 45 patients with an ordinary low-pitched rumble. Phonocardiography, and M-mode and two-dimensional (2-D) echocardiography were performed in all patients. Color and continuous wave Doppler echocardiography were performed in four patients with a high-pitched rumble and in 13 with a low-pitched rumble. Results obtained were as follows: 1. Points of the maximum intensity of a rumble: A low-pitched rumble was best heard at the apex in all patients. A high-pitched rumble was best heard at the mesoapical area, except in one patient who had an oval mitral orifice by 2-D echocardiography. 2. Correlation between the pitch of rumble and the shape of the mitral orifice: In five of the six patients with a high-pitched rumble, the mitral orifice had a tadpole-shaped deformity, in which commissural fusion and valvular thickening were more marked anterolaterally than posteromedially. Among 45 patients with oval, slit or pinhole-like valve orifices, only one had a high-pitched rumble. 3. Direction of the left ventricular (LV) inflow jet as observed by color Doppler echocardiography: On the short-axis view at the level of the papillary muscles, the inflow jet was directed toward the medial portion of the LV cavity in the patients with a high-pitched rumble. However, it was directed towards the central portion of the LV cavity in all patients with a low-pitched rumble. On the apical long-axis view, no distinct difference was detected in the direction of the LV inflow jet between the two groups. 4. Other findings: There were no significant differences between the two groups in the mitral orifice area, the peak velocity of LV inflow, fractional shortening of the LV, dimension of the left atrium, Wells' index and the degree of organic change in the subvalvular structures. These results suggest that the deformity of the mitral valve and resultant changes in the direction of the LV inflow jet may play an important role in the mechanism of producing a high-pitched diastolic rumble in mitral stenosis.

[A case of mitral valve prolapse simulating mitral tumor or vegetation].

We observed a case of mitral valve prolapse (MVP) mimicking mitral tumor or vegetation. The patient was a 34-year-old man. He complained of palpitation and precordial oppression. Shaggy and mass echoes of the anterior mitral valve were recorded by M-mode and two-dimensional echocardiograms. The "mass" echo was confirmed to by originated in a markedly myxomatous change of the spongiosa layer of the mitral valve at operation. These problems on echocardiographic diagnosis of MVP and mitral "mass", such as mitral tumor or vegetation, were discussed in detail.

New enzymatic determination of sialic acid in serum.

This enzymatic method for determination of sialic acid involves use of neuraminidase (EC 3.2.1.18), N-acetylneuraminate lyase (EC 4.1.3.3), acylglucosamine 2-epimerase (EC 5.1.3.8), N-acetylhexosamine oxidase (from Pseudomonas sp.), and peroxidase (EC 1.11.1.7). Because the method does not require pyruvic acid in the assay medium, interference by pyruvic acid in serum can be avoided. This simple, accurate assay is little affected by other substances in serum.

Antagonistic effects of alpha-adrenoceptor blocking agents on reticuloruminal hypomotility induced by xylazine in cattle.

The intravenous injection of a standard dose (0.05 mg/kg) of xylazine inhibited reticuloruminal motility in cattle. Pretreatment with adrenoceptor antagonists showing alpha 2-blocking activity, tolazoline (0.5 mg/kg) and yohimbine (0.2 mg/kg), antagonized the xylazine-induced reticuloruminal amotility. Tolazoline was more effective than yohimbine, since an antagonistic effect was not seen at 0.5 mg/kg yohimbine, and yohimbine at 0.2 mg/kg was less effective than tolazoline at 0.5 mg/kg. An adrenoceptor antagonist showing alpha 1-blocking activity, prazosin, did not prevent the inhibition of reticuloruminal motility by xylazine. The xylazine-induced reticuloruminal amotility was also not prevented by either a dopamine receptor antagonist, domperidone, or an opiate receptor antagonist, naloxone. These results suggest that xylazine inhibits bovine reticuloruminal motility through its activation of alpha 2-adrenoceptors, and show that tolazoline can be used as a specific antagonist of xylazine in studies of the alpha-adrenergic influence on reticuloruminal motility in cattle.

[Genesis and clinical significance of the "low-pitched" aortic ejection sound].

The genesis and clinical significance of the aortic ejection sound with a low-frequency predominance and delayed appearance were studied. This is recorded on the phonocardiogram in some patients with left ventricular dysfunction. Subjects studied consisted of 10 patients with a low-pitched ejection sound and seven patients with an ordinary high-pitched aortic ejection sound. No patients had echocardiographic findings suggestive of organic lesions of the aortic valve. Time relationships among the ejection sounds, aortic valve echograms and carotid artery pulses, and then movements of the aortic valve cusps and non-invasively estimated left ventricular systolic function were compared between the two groups. Results were as follows: 1. The low-pitched ejection sound: 1) The beginning of the sound was nearly coincident with the onset of the upstroke of the carotid artery pulse and the initial full opening of the aortic valve cusps. 2) The beginning of the ejection systolic murmur followed immediately after the ejection sound. 3) The amplitude of the sound was closely related to the height of the carotid artery pulse in a case of atrial fibrillation. 2. The low-pitched ejection sound vs the high-pitched ejection sound: 1) The onset of the low-pitched ejection sound was significantly delayed. 2) The amplitude and the velocity of the initial opening of the aortic valve cusps were significantly decreased. 3) The preejection period (PEP) was significantly prolonged; the ejection time (ET) was significantly shortened; and the PEP/ET ratio was significantly increased.(ABSTRACT TRUNCATED AT 250 WORDS)

[Systolic clicks in mitral valve prolapse: their pathophysiological relationship to the grade and causes of prolapse].

To re-evaluate the clinical significance of non-ejection systolic clicks in mitral valve prolapse (MVP), 154 patients with idiopathic MVP (idiopathic group) and 54 patients with secondary MVP associated with atrial septal defect (secondary group) were studied using phonocardiography and two-dimensional echocardiography. There was no significant difference in the incidence of systolic clicks between the idiopathic (30%) and secondary (24%) groups. The severer the degree of prolapse, the higher the incidence of systolic clicks in both the groups. A systolic click was observed independently of age in the idiopathic group. In the secondary group, however, a systolic click was not observed in patients under 30 years old and increased with advancing age in frequency. The intensity of systolic click tended to increase in proportion to the increase of severity of prolapse in both the groups. These results suggested that systolic clicks may appear irrespective of the causes of prolapse and that the production and the intensity of systolic clicks may be related to the degree of changes in the mitral valve-chordal system and the degree of prolapse.