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Transcervical artificial insemination in sheep: effects of a new transcervical artificial insemination instrument and traversing the cervix on pregnancy and lambing rates.

Cervical anatomy limits the use of transcervical intrauterine artificial insemination (TC AI) in sheep. We have developed an instrument to cope atraumatically with the cervix; although this instrument has not affected fertilization rate or pregnancy rate through Day 3, the effects on sperm transport and pregnancy after Day 3 are not known. The objective of the present study was to determine whether our TC AI instrument affected sperm transport, pregnancy rates, or lambing rate. In Experiment 1, ewes were assigned to two treatments: TC AI using the new TC AI instrument (n=10) or AI via laparotomy using a laparoscopic AI instrument (n=10). Twenty hours after artificial insemination, the uterine horns and oviducts were recovered and flushed to collect spermatozoa. Sperm transport did not differ (P>0.05) between the two treatments. In Experiment 2, ewes were assigned to three treatments: TC AI using the new TC AI instrument+sham intrauterine AI via laparotomy (n=29); sham TC AI+intrauterine AI via laparotomy using a laparoscopic AI instrument (n=29); and sham TC AI+intrauterine AI via laparotomy using the new TC AI instrument (n=30). On Day 14 after AI, uteri were collected and flushed to recover blastocysts. Transcervical deposition of semen reduced (P<0.05) Day 14 pregnancy rate (17.2% versus 61%), but intrauterine deposition of semen using the TC AI instrument via midventral laparotomy increased (P<0.05) Day 14 pregnancy rate (76.6% versus 44.8%). In Experiment 3, ewes were assigned to two treatments: sham cervical manipulation (n=40) or cervical manipulation to mimic TC AI (n=40). Immediately after treatment, each ewe was mated with a ram and watched until the ram mounted and ejaculated into the ewe. Treatment did not affect Day 30 or 50 pregnancy rate (67.5 and 66.2%, respectively), determined ultrasonically, or lambing rate (62.5%). The differences between Days 30 and 50 pregnancy rates and lambing rate were not significant. In Experiment 4, ewes were assigned to two treatments: TC AI (n=99) or laparoscopic AI (n=99). Transcervical AI reduced (P<0.01) Day 30 (TC AI versus laparoscopic AI; 5.0% versus 46.0%) and Day 50 pregnancy rates (4.0% versus 41.0%), determined ultrasonically, and lambing rate (4.0% versus 41.0%). Although the TC AI procedure significantly reduced pregnancy and lambing rates, large numbers of spermatozoa deposited at natural insemination seemed to compensate. Because our TC AI procedure has all but eliminated any visual evidence of trauma, and because the procedure does not seem to affect sperm transport or embryonal survival until Day 3, we speculate that cervical manipulation associated with TC AI may activate pathways that interrupt pregnancy between Days 3 and 14.

Animals↗

Development of a new transcervical artificial insemination method for sheep: effects of a new transcervical artificial insemination catheter and traversing the cervix on semen quality and fertility.

The difficulty of traversing the cervix severely limits transcervical artificial insemination (TC AI) in sheep. Cervical trauma and poorly designed instruments can reduce fertility after AI. To overcome problems associated with TC AI, we developed a new TC AI catheter. Three bench experiments were conducted to determine the effects of the new TC AI catheter on semen quality independent of the effects of moving the catheter through the cervix. In each of the three bench experiments, the standard laparoscopic instrument for intrauterine AI in sheep was used as the control for the TC AI catheter. In Experiment 1, the total volume of semen extender expelled and void volumes for both types of AI instrument (TC versus laparoscopic) were determined. In Experiment 2, the effects of each type of AI instrument (TC versus laparoscopic) on semen quality, estimated as percentage motility and percentage forward progressive motility, of frozen-thawed semen was determined. In Experiment 3, the effects of both types of AI instrument (TC versus laparoscopic) on number of spermatozoa expelled was determined. The type of AI instrument affected neither semen quality nor the number of spermatozoa expelled. However, void volume differed (P < 0.01) between the two instruments. After differences in void volume were taken into account, an in vivo experiment was conducted to determine whether using our new TC AI catheter for TC or surgical intrauterine AI affected fertilization and pregnancy rates. For this, ewes were assigned to one of three treatments: (1) TC AI using the new TC AI catheter + sham AI via laparotomy (n = 9); (2) sham TC AI + AI via laparotomy using a laparoscopic AI instrument (n = 8); and (3) sham TC AI + AI via laparotomy using the new TC Al catheter (n = 10). To synchronize estrus, progestogenated pessaries were inserted and left in place for 12 days. On Day 5 after pessary insertion, PGF2alpha (15 mg) was given i.m. At pessary removal, 400 IU of eCG were administered i.m. Ewes were inseminated 48-52 h after pessary removal using fresh diluted semen (200 x 10(6) to 350 x 10(6) spermatozoa per 0.2 ml) pooled from the same four rams each day during the experiment. At 72 h after AI, uteri were collected postmortem and flushed. Oocytes and embryos were recovered and evaluated. Treatments did not affect (P > 0.01) ovum and embryo recovery rate (mean = 87.3%), fertilization rate (59.3%), or Day 3 pregnancy rate (mean = 66.6%). We conclude from these data that the use of our new TC AI catheter for TC AI or intrauterine AI should not impair the success of AI in sheep.

Animals↗

Effect of time of insemination on number of accessory sperm, fertilization rate, and embryo quality in nonlactating dairy cattle.

Two experiments were conducted to determine the effect of insemination time on number of accessory sperm per embryo (ovum), fertilization rate, and embryo quality. Semen was collected from three fertile Holstein bulls and cryopreserved in egg yolk-citrate-glycerol. In experiment 1, cows were continuously monitored for behavioral estrus by the HeatWatch estrous detection system and were artificially inseminated (AI) with one 0.5-ml straw (25 x 10(6) sperm) at the onset of estrus (AI 0 h), 12 h after onset (AI 12 h), or received natural service at 0 h (Nat 0 h) from one of three bulls. From 150 inseminations, 115 embryos and ova (AI 0 h: n = 39; AI 12 h: n = 39; Nat 0 h: n = 37) were recovered 6 or 7 d after insemination. Fertilization rates differed between treatments (AI 0 h: 67%; AI 12 h: 79%; Nat 0 h: 98%). Median accessory sperm per embryo (ovum) also differed (AI 0 h: 1; AI 12 h: 10; and Nat 0 h: 27) and paralleled the fertilization rate. Embryo quality was not affected by insemination time or natural service. In experiment 2, cows received AI at 0, 12, or 24 h (AI 24 h) after the onset of estrus as determined by HeatWatch. From 154 inseminations, 117 embryos and ova (AI 0 h: n = 39; AI 12 h: n = 39; AI 24 h: n = 39) were recovered 6 or 7 d after insemination. Fertilization rates did not differ in experiment 2 (AI 0 h: 66%; AI 12 h: 74%; AI 24 h: 82%); however, a trend toward a higher fertilization rate accompanied AI 24 h. Median accessory sperm values increased from AI 0 h (1) to AI 24 h (4). Embryo quality declined with AI at increasing intervals after onset of estrus, as percentages of excellent and good, fair and poor, and degenerate embryos were as follows: 77, 15, 8; 52, 38, 10; and 47, 19, 34 for the 0-, 12-, and 24-h inseminations, respectively. Results indicate AI 12 h after the onset of estrus provides a compromise between potential fertilization failure (AI 0 h) and embryo failure (AI 24 h), despite increased accessory sperm per embryo (ovum) after AI 24 h. Artificial insemination 12 h after onset of estrus should optimize fertility of dairy cattle through an acceptable fertilization rate, number of accessory sperm per embryo, and desirable embryo quality.

Animals↗

Trucking stress at breeding does not lower conception rate of beef heifers.

The goal of this study was to determine whether 1 h of trucking stress before or after artificial insemination (AI) altered the conception rate of beef heifers. Estrus was synchronized in heifers with prostaglandin F(2alpha). The 3 treatment groups consisted of 1) AI (control heifers, n = 93); 2) Truck + AI (trucked for 1 h immediately before AI, n = 81); and 3) AI + Truck (trucked for 1 h immediately after AI, n = 82). All heifers were artificially inseminated by a single technician with semen from a single ejaculate. Blood samples were collected for cortisol measurement 1 h before AI, immediately before and after AI, and 1 h after AI in the AI (n = 6), Truck + AI (n = 9), and AI + Truck (n = 8) groups Pregnancy in heifers was confirmed either at slaughter or by palpation per rectum. Trucking before AI elevated (P < 0.01) serum cortisol concentrations. Artificial insemination alone increased (P < 0.01) serum cortisol concentrations in AI heifers. The increase in serum cortisol concentrations caused by trucking after AI was not significant (P > 0.05). Areas under the cortisol curves in Truck + AI heifers are greater (P < 0.05) than in AI heifers. The conception rates of AI heifers (50.5%), Truck + AI heifers (51.9%) and AI + Truck heifers (58.5%) are not different (P > 0.05). This study demonstrates that 1 h of trucking stress either before or after AI did not lower the conception rate of heifers.

Journal Article↗

The effect of intravaginal applied GnRH-agonist on the time of ovulation and subsequent reproductive performance of weaned multiparous sows.

In order to prove the effect of 'fixed time insemination' and insemination at standing oestrus after post-weaning application of GnRH, in a Croatian large breeding unit, 502 sows were assigned to three groups and were artificially inseminated (AI) at their first post-weaning oestrus as many times as they stand, in 24-h intervals. The groups were treated as follows: group 1 (control, n = 160) were AI during their standing reflex; group 2 ['GnRH-fixed time insemination' (GnRH-FT-AI), n = 175] were AI, independent of detection of oestrus and following administration of GnRH-agonist at 96 h post-weaning; group 3 [GnRH insemination at standing oestrus (GnRH-OE-AI), n = 167] the animals were GnRH-agonist treated as group 2 and were AI at their standing reflex. Pre-trial daily average lactational feed intake, average daily feed intake from weaning to oestrus, oestrus within 6 days post-weaning (%), ovulation within 6 days post-weaning (%), weaning-to-oestrus interval (h), duration of oestrus (h), follicle size (mm), interval from oestrus to ovulation (h), subsequent day 24 pregnancy rate (%), farrowing rate (%) and total pigs born were evaluated. Pre-trial average daily lactational voluntary feed intake was 7.1 +/- 0.08 kg in group 1, 7.0 +/- 0.07 kg in group 2 and 7.1 +/- 0.17 kg in group 3 (p > 0.05). Average voluntary daily feed intake from weaning to oestrus was 5.1 +/- 0.3 kg in group 1, 5.2 +/- 0.5 kg in group 2 and 5.2 +/- 0.19 kg in group 3 (p > 0.05). Oestrus was detected within 6 days post-weaning in 134 (83.8%) in control, 164 (93.7%) in GnRH-FT-AI and 155 (92.8%) animals in GnRH-OE-AI groups (p = 0.05). Follicle size did not differ (p > 0.05) among the groups. In control 82.8%, in GnRH-FT-AI 91.5% and in GnRH-OE-AI 91.0% of the sows ovulated within 6 days post-weaning (p = 0.04), and had 80.6, 90.9 and 89.7% 24-day pregnancy rates (p = 0.16), respectively. In GnRH-FT-AI group 90.2%, in GnRH-OE-AI sows 89.7%, in control animals 79.9% farrowing rates were recorded (p = 0.17). Weaning to oestrus interval was 113.1 h in control, 114.1 h in GnRH-FT-AI and 112.6 h GnRH-OE-AI (p > 0.05). Duration of oestrus was significantly shorter in GnRH-FT-AI (44.9 h) and GnRH-OE-AI (48.1 h) animals, compared with the control (62.9 h) sows (p = 0.001). Similarly, the interval from oestrus to ovulation revealed significant (p = 0.004) differences between the groups (control 44.1 h, GnRH-OE-AI 34.1 h and GnRH-FT-AI 32.9 h). GnRH-FT-AI (12.5) and GnRH-OE-AI (12.6) sows had significantly higher (p = 0.01) number of total pigs born (n = 10.4) compared with control sows. GnRH-agonist-gel treatment to the sow shortens duration of oestrus, the interval from oestrus to ovulation, and may eliminate the need for oestrus detection in the hands of skilled personnel.

Administration, Intravaginal↗

Cortical synthesis of azimuth-sensitive single-unit responses with nonmonotonic level tuning: a thalamocortical comparison in the cat.

1. Azimuth and sound pressure level (SPL) tuning to noise stimulation was characterized in single-unit samples obtained from primary auditory cortex (AI) and in areas of the medial geniculate body (MGB) that project to AI. The primary aim of the study was to test the hypothesis that AI is an important site of synthesis of single-unit responses that exhibit both azimuth sensitivity (tendency for directionally restricted responsiveness) and nonmonotonic (NM) level tuning (tendency for decreased responsiveness with increasing SPL). This was accomplished by comparing the proportions of such responses in AI and MGB. 2. Samples consisted of high-best-frequency (BF) single units located in MGB (n = 217) and AI (n = 216) of barbiturate-anesthetized cats. The MGB sample was obtained mainly from recording sites located in two nuclei that project to AI, the ventral nucleus (VN, n = 118) and the lateral part of the posterior group of thalamic nuclei (Po, n = 84). In addition, a few MGB units were obtained from the medial division (n = 8) or uncertain locations (n = 7). Each unit's responses were studied using noise bursts presented from azimuthal sound directions distributed throughout 180 degrees of the frontal hemifield at 0 degrees elevation. SPL was varied over an 80-dB range in steps of < or = 20 dB at each location. Similarities and differences in azimuth and level tuning were evaluated statistically by comparing the AI sample with the entire MGB sample. If they were found to differ, the AI, VN, and Po samples were compared. 3. Azimuth function modulation was used as a measure of azimuth sensitivity, and its mean was greater in AI than in MGB. NM strength was defined as the percentage reduction in level function value at 75 dB SPL and its mean was greater in AI (showing a greater tendency for decreased responsiveness) than in MGB. Azimuth-sensitive (AS) NM units were identified by jointly categorizing each sample according to both azimuth sensitivity (sensitive and insensitive categories) and NM strength (NM and monotonic categories). AS NM units were much more common in the AI sample than in any of the MGB samples, suggesting that some such responses are synthesized in AI. 4. A vast majority of AI NM units have been reported to be AS, showing a preferential association (linkage) between these two response properties. This finding was confirmed in AI, but was not found to be the case in MGB. This suggests that a linkage between these response properties emerges in the cortex, presumably as a result of synthesis of NM AS responses. Although the functional significance of the linkage is unknown, NM responses may reflect excitatory/inhibitory antagonism that provides AS AI neurons with sensitivity to stimulus features beyond that which is present in MGB. 5. Breadth of azimuth tuning of AS cells was measured as the portion of the frontal hemifield over which azimuth function values were > 75% of maximum (preferred azimuth range, PAR). PARs were broadly distributed in each structure, and mean PAR was narrower in AI than in MGB. A preferred level range (PLR) was defined for NM level functions as the range over which values were > 75% of maximum, and mean PLRs were similar in each sample. There was a weak, but significant, positive correlation between PARs and PLRs in AI but not in MGB. This further suggests a linkage between azimuth and level tuning in AI that does not exist in MGB. 6. Best azimuth (midpoint of the PAR) was used to classify cells as contralateral preferring, ipsilateral preferring, midline preferring, or multipeaked. Samples from AI and MGB exhibited similar distributions of these categories. Contralateral-preferring cells represented a majority of each sample, whereass midline-preferring, ipsilateral-preferring, and multipeaked cells each represented smaller proportions. This suggests that the azimuth preference distribution in AI largely reflects that in MGB. 7. A best SPL was defined as the midpoint of the PLR. This wa

Animals↗

Prognostic significance of apoptotic index in completely resected non-small-cell lung cancer.

PURPOSE: To evaluate the significance of apoptotic index (AI) as a prognostic factor after surgery for non-small-cell lung cancer (NSCLC). PATIENTS AND METHODS: A total of 236 patients who underwent surgery for previously untreated pathologic stage I to IIIa NSCLC between 1985 and 1990 were reviewed. AI was defined as the number of apoptotic cells, detected by terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate-biotin nick end-labeling, per 1,000 tumor cells. Proliferative index (PI) and aberrant p53 expression were also evaluated immunohistochemically. RESULTS: The 5-year survival rate for the lowest-AI group (AI < 5.0) was 74.7%; those for the lower-AI group (5.0 < or = AI < 11.0) and the higher-AI group (11.0 < or = AI < 25.0) were 51.6% and 57.8%, respectively. These survival rates were significantly lower than that of the lowest-AI group (P =.021 and P =.043, respectively). The highest-AI group (25.0 < or = AI), however, showed the most favorable prognosis, with a 5-year survival rate of 83.2%. Multivariate analysis confirmed that a moderate AI (5.0 < or = AI < 11.0 or 11.0 < or = AI < 25.0) was a significant factor to predict poor prognosis. The PIs for the lowest-, the lower-, the higher-, and the highest-AI groups were 32.3%, 48.0%, 54.3%, and 50.7%, respectively. The lowest-AI group showed a favorable prognosis because of its low PI, whereas the lower- and the higher-AI groups had a poor prognosis caused by increased cancer-cell proliferation. The highest-AI group showed the most favorable prognosis because apoptotic cell death overcame cell proliferation. No significant correlation was observed between AI and aberrant p53 expression. CONCLUSION: AI proved to be an independent prognostic factor in NSCLC.

Adenocarcinoma↗

Double insemination and gonadotropin-releasing hormone treatment of repeat-breeding dairy cattle.

Our objective was to determine if double inseminations during the same estrous period of dairy cattle eligible for their third or fourth service (repeat breeders) would improve pregnancy rates equivalent to injections of GnRH given at the time of AI. Repeat-breeding, lactating cows from six herds (five herds in the San Joaquin Valley of central California and one herd in northeast Kansas) were assigned randomly to four treatment groups when detected in estrus: 1) single AI plus no injection, 2) single AI plus 100 micrograms GnRH at AI, 3) double AI plus no injection, or 4) double AI plus 100 micrograms of GnRH at AI. Inseminations were performed according to the a.m.-p.m. rule. The second AI for the double AI treatment was given 12 to 16 h after the first AI. Injections of GnRH were given intramuscularly immediately following the single AI or the first AI of the double AI. Pregnancy rates of cows given a single AI and hormone injection were numerically higher in all six herds than those of their herdmates given only a single AI. In five of six herds, the pregnancy rates of cows given a double AI and hormone injection were numerically higher than pregnancy rates of their herdmates given only a double AI. Overall pregnancy rates for the four treatments were 1) 112/353 (32.1%), 2) 165/406 (41.6%), 3) 119/364 (33.5%), and 4) 135/359 (37.5%). Gonadotropin-releasing hormone increased pregnancy rates of repeat breeders compared with controls given only a single AI. No further benefit beyond the single AI was accrued from the double AI treatment, with or without concurrent hormone administration.

Animals↗

Diagnosis of misery perfusion using noninvasive 15O-gas PET.

UNLABELLED: To avoid arterial blood sampling and complicated analyses in 15O-gas PET studies, we evaluated a noninvasive technique using the count-based method for measuring asymmetric increases in oxygen extraction fraction (OEF) in cerebrovascular disease. METHODS: Eighteen patients (mean age +/- SD, 61 +/- 16 y) with atherothrombotic large-cerebral-artery disease were studied for the measurement of hemodynamic parameters using the 15O-gas steady-state method with inhalation of 15O2, C15O2, and C15O. All patients also underwent H2(15)O PET with the bolus injection method. Count-based ratio images of 15O2/C15O2 and (15)O2/H2(15)O were calculated, and asymmetry indices (AIs) were obtained (cbOEF(SS)-AI and cbOEF(BO)-AI, respectively) using regions of interest drawn bilaterally on the cerebral cortices. These AIs were compared with the AIs of absolute OEF (qOEF-AI) and with those after cerebral blood volume (CBV) correction. A contribution factor for this correction was defined as a variable alpha, and the effect of the correction was evaluated. RESULTS: cbOEF(SS)-AI underestimated qOEF-AI significantly, especially with a greater AI (P < 0.05). cbOEF(BO)-AI linearly correlated well with qOEF-AI. CBV correction improved the slopes of regression lines between qOEF-AI and cbOEF(SS)-AI, and the optimal alpha was defined as 0.5. On the other hand, cbOEF(BO)-AI fairly estimated qOEF-AI without CBV correction. Correlation between qOEF-AI and cbOEF(BO)-AI was adversely affected, and the mean bias was increased, with a greater alpha. CONCLUSION: cbOEF(BO)-AI can fairly estimate the AI of OEF without CBV correction, whereas cbOEF(SS)-AI might require CBV correction for better estimation. The examination time and stress to patients would be reduced with the count-based method because it is noninvasive.

Aged↗

Fertility after deep intra-uterine artificial insemination of concentrated low-volume boar semen doses.

UNLABELLED: Boar semen can be successfully frozen - highly packed - in small containers (medium-straw, MS or MiniFlatPack, MFP). The use of deep intra-uterine artificial insemination (DIU-AI) can make possible the deposition of small volumes of this thawed, non re-extended semen deeply intra-uterine, close to the sperm reservoir. The present experiments studied the fertility achieved after single or double DIU-AI per oestrus, with special attention to the interval between AI and spontaneous ovulation. Semen from two boars of proven fertility was frozen in MS or MFP holding 1 x 10(9) total spermatozoa. Multiparous (2-5 parity, n=42) crossbred sows were checked for oestrous behaviour after weaning and the occurrence of spontaneous ovulation was checked with transrectal ultrasonography (TUS) to establish the mean interval between onset of oestrus (OO) and ovulation which was found to be when approximately 2/3 of the oestrus period has passed. The sows were, in the following standing oestrus, subjected to DIU-AI using thawed semen from either MS (n=20) or MFP (n=22), inseminated without further re-extension. The sows were randomly allotted to one of three groups: (1) single DIU-AI 8 h before expected ovulation (control group, n=19); (2) single DIU-AI 4 h before expected ovulation (treatment group S, n=15); and (3) double DIU-AI 12 and 4 h before expected ovulation (treatment group D, n=8). Occurrence of spontaneous ovulation was confirmed by TUS, performed as during the first oestrous period and used to determine the real interval of DIU-AI and ovulation. Pregnancy was also confirmed by TUS 28 days after OO in those sows not returning to oestrus. These sows were slaughtered (30-45 days of pregnancy), and the appearance of the reproductive tract and ovaries, the number of live and dead foetuses, of implantation sites and of corpora lutea (CL) were recorded. Sows (n=9) returning to oestrus ("open") were re-inseminated (either once [n=4] or twice [n=5]) the following oestrus with either MFP (n=5) or MS (n=4) and slaughtered 12-14 h post-ovulation for recovery of tubal oocytes and of spermatozoa from the uterotubal junctions (sperm reservoir), to assess the degree of effectiveness of sperm transport. Post-thaw sperm motility was 44.3+/-3.21% in MFP and 42.8+/-0.72% for MS (LSmean+/-S.E.M., n.s.), and did not significantly change from thawing to AI. The DIU-AI could be performed in all sows, but insertion was difficult (slow >5 min) in 5/42 sows. Four of these sows returned to oestrus. Pregnancy rate averaged 35% (group D: 25%, group S: 40%, control: 36%, n.s.). The interval between DIU-AIs and spontaneous ovulation varied largely, ranging from -13 to -3 h for group C, for group S from -11 to +3 h and for group D from -17 to -4 h. Pregnancy rates were clearly related to the interval DIU-AI and ovulation, being highest (60%, 12/20) when AI occurred between 8 and 4 h before spontaneous (not expected) ovulation. The number of implantation sites ranged 6-22 (n.s. among groups), and the number of alive foetuses 2-11 (n.s. among groups). Implantation rate (total number of implantations/CL) ranged 48.0-69.7% being highest in the D-group (P<0.05). The examination of the "open" sows slaughtered 12-14 h post-ovulation revealed few recovered oocytes were fertilized (approximately 10%). Only 40% of oocytes had spermatozoa bound to the zona pellucida, not more than two spermatozoa per oocyte. Moreover, low sperm numbers (approximately 4000) were found in the sperm reservoirs (UTJs), irrespective of using single or double DIU-AI (n.s.). The highest values (P<0.05) for these variables were recorded when DIU-AI (either single or double [second AI]) occurred 4-8 h before ovulation, especially when MFP-semen was used (P<0.05). IN CONCLUSION: (1) DIU-AI can be easily performed in most sows; (2) pregnancies can be obtained by the DIU-AI of low volumes of highly concentrated frozen-thawed boar semen, once or twice during oestrus, but fertility is still low, probably owing to an unsatisfactory sperm transport when expected and real ovulation differ; and (3) fertility is related to the interval DIU-AI and ovulation which should be -8 to -4 h of spontaneous ovulation and to the package, MFP having shown better results in vivo. The results stress the need for careful, and frequent, control of oestrus signs.

Animals↗

Deferoxamine test and bone disease in dialysis patients with mild aluminum accumulation.

Aluminum bone disease is a frequent complication of dialysis patients. The deferoxamine (DFO) test has been advocated as a noninvasive procedure for the diagnosis of AI bone lesion. However most of these studies have been performed in symptomatic patients with significant AI bone disease. Whether this test may provide similar data at an earlier stage of AI toxicity is not known. The present study evaluates prospectively 28 patients with mild AI load. Patients studied ranged in age from 21 to 65 years; duration of dialysis was 5.6 +/- 3.2 years; deferoxamine, 40 mg/kg body weight, was infused at the end of dialysis. Serum AI was measured before DFO administration and before the next dialysis treatment. Bone biopsies were performed in all patients. Cortical bone AI was determined biochemically; trabecular and cortical bone AI were also determined histochemically. Mean basal serum AI (43.2 +/- 30.8 micrograms/L) and cortical bone AI (25.7 +/- 35.2 micrograms/g) were moderately increased. Basal serum AI correlated (r = 0.77) with the increment in serum AI after DFO infusion. After DFO, stainable trabecular and cortical bone AI correlated in a similar manner with both basal serum AI and increment in serum AI. Only biochemically determined cortical bone AI was not significantly related to basal serum AI. Nineteen of the 28 patients had evidence of osteitis fibrosa on bone biopsy. Stained AI surfaces but not trabecular AI were different in patients with low and patients with high bone formation rates. The bone findings, assessed as bone formation rates and resorption surfaces, did not correlate with biochemically or histochemically determined bone AI.(ABSTRACT TRUNCATED AT 250 WORDS)

Aluminum↗

Reduced atherosclerotic lesions in mice deficient for total or macrophage-specific expression of scavenger receptor-A.

The absence of the scavenger receptor A (SR-A)-I/II has produced variable effects on atherosclerosis in different murine models. Therefore, we examined whether SR-AI/II deficiency affected atherogenesis in C57BL/6 mice, an inbred strain known to be susceptible to diet-induced atherosclerotic lesion formation, and whether the deletion of macrophage SR-AI/II expression would modulate lesion growth in C57BL/6 mice and LDL receptor (LDLR)(-/-) mice. SR-AI/II-deficient (SR-AI/II(-/-)) female and male mice on the C57BL/6 background were challenged with a butterfat diet for 30 weeks. No differences were detected in plasma lipids between SR-AI/II(-/-) and SR-AI/II(+/+) mice, whereas both female and male SR-AI/II(-/-) mice had a tremendous reduction (81% to 86%) in lesion area of the proximal aorta compared with SR-AI/II(+/+) mice. Next, to analyze the effect of macrophage-specific SR-AI/II deficiency in atherogenesis, female C57BL/6 mice were lethally irradiated, transplanted with SR-AI/II(-/-) or SR-AI/II(+/+) fetal liver cells, and challenged with the butterfat diet for 16 weeks. In a separate experiment, male LDLR(-/-) mice were reconstituted with SR-AI/II(-/-) or SR-AI/II(+/+) fetal liver cells and challenged with a Western diet for 10 weeks. No significant differences in plasma lipids and lipoprotein profiles were noted between the control and experimental groups in either experiment. SR-AI/II(-/-)-->C57BL/6 mice, however, had a 60% reduction in lesion area of the proximal aorta compared with SR-AI/II(+/+)-->C57BL/6 mice. A similar level of reduction (60%) in lesion area was noted in the proximal aorta and the entire aorta en face of SR-AI/II(-/-)-->LDLR(-/-) mice compared with SR-AI/II(+/+)-->LDLR(-/-) mice. These results demonstrate in vivo that SR-AI/II expression has no impact on plasma lipid levels and that macrophage SR-AI/II contributes significantly to atherosclerotic lesion formation.

Animals↗

Transcervical artificial insemination of Australian Merino ewes with frozen-thawed semen.

We compared conventional methods for laparoscopic and cervical artificial insemination (AI) to a transcervical AI procedure (Guelph System for Transcervical AI; GST-AI) for use with frozen semen in Merino ewes. The GST-AI procedure was performed by an experienced operator in Experiment 1 (771 ewes) and by 2 inexperienced operators in Experiment 2 (555 ewes). In Experiment 1, intrauterine insemination by GST-AI was achieved in 76% of the ewes. The pregnancy rate at Day 70 for ewes inseminated by laparoscopy (48%, 120 251 ) was higher (P<0.01) than for ewes inseminated by either intrauterine GST-AI (32%, 64 201 ) or cervical AI (9%, 24 256 ). The overall (intrauterine and intracervical) pregnancy rate for GST-AI was 26% (68 264 ) and was unaffected by depth of insemination within the cervix. Pregnancy rates were unaffected by ram or day of insemination. In Experiment 2, the operators achieved intrauterine inseminations by GST-AI in 43% (78 182 ) of the ewes, with a significant operator effect (P<0.01) on depth of cervical penetration. The pregnancy rate to intrauterine GST-AI (40%, 31 78 ) did not differ from that to laparoscopic insemination. The total pregnancy rate for GST-AI in Experiment 2 (19%, 34 182 ) was lower (P<0.05) than that for laparoscopic AI (39%, 72 187 ) but superior (P<0.05) to that for cervical AI (1%, 1 186 ). The GST-AI pregnancy rates were affected by depth of AI (P<0.01) and by operator (P<0.05). It is concluded that GST-AI is superior to cervical AI, and may have application in Merinos if cervical penetration rates can be improved.

Journal Article↗

Comparison of fertility data from vaginal vs intrauterine insemination of frozen-thawed dog semen: a retrospective study.

Fertility data from 327 artificial inseminations (AIs) using frozen-thawed dog semen are presented here. The AIs were performed in 274 bitches using semen from 185 males of 76 breeds. The data cover all AIs conducted during 1983 through 1995 at Cryogenetic Laboratories (CLONE) in the United States with AKC-registered and research bitches, and all AIs carried out at the Department of Obstetrics and Gynecology at the Swedish University of Agricultural Sciences in Uppsala, Sweden, using semen frozen by CLONE, in 0.5-mL straws. Semen was frozen using a standardized, three-step liquid nitrogen vapor freezing method. Whelping rates > 70% were obtained when post-thaw motility was 40% or higher. The inseminations were made either directly into the uterus using transcervical catheterization with the Norwegian catheter (NIU; 167 AIs) or a fiberoptic endoscope (EIU; 19 AIs), or in the cranial vagina (VAG; 141 AIs). Resulting whelping rates were 84.4% (NIU), 58.9% (VAG; P < 0.001), and 57.9% (EIU). Increasing the number of VAG AIs per cycle from 1 to 2 enhanced the whelping rate (P < 0.05). The mean interval from the first AI to whelping was 61.8 +/- 2.4 d, and was longer for VAG AIs (62.7 +/- 2.7 d) than for NIU AIs (61.2 +/- 2.1 d; P < 0.001). The mean interval from the last AI was 60.1 +/- 1.9 d, and did not differ between VAG AIs (60.2 +/- 2.2 d) and NIU AIs (60.0 +/- 1.6 d). Gestation length was not influenced by breed or litter size. A total of 1158 pups resulted from the 327 AIs. Litter size was 5.4 +/- 3.0 (NIU), 4.0 +/- 2.7 (VAG; P < 0.001), and 6.0 +/- 2.1 (EIU). Litter size was also influenced by breed (P = 0.006) and, for VAG AIs, by the number of inseminations performed per cycle (P = 0.009). This study is the largest that has been carried out on frozen-thawed dog semen AI. It shows that using a good method for cryopreservation, together with nonsurgical intrauterine AI employing the Norwegian catheter, can yield whelping rates and litter sizes similar to those reported from well-controlled natural matings. Furthermore, this is the first study to show that intrauterine deposition of frozen-thawed dog semen results in a significantly higher whelping rate and larger litter size than vaginal deposition.

Animals↗

A preliminary retrospective treatment and pharmacoeconomic analysis of asthma care provided by allergists, immunologists, and primary care physicians in a teaching hospital.

Allergy immunology specialists (AIs) differ from primary care physicians (PCP) in their treatment of asthma. A limited retrospective chart review of several visits over a 1-year period in 1997 evaluating the quality of asthma care by AIs vs. PCPs was conducted in an academic center. Data concerning quality, effectiveness and cost of asthma care was randomly collected from 15 AIs and 15 PCPs from charts at 3-month intervals over a 1-year period. Information obtained from data collection forms revealed that asthma patients evaluated by AIs had more visits and received a greater quantity of medication compared to those treated by PCPs. All 15 patients with persistent asthma followed by AIs were treated with inhaled corticosteroids at each visit in contrast to only 80% of those treated by PCPs. The total numbers of controller medications (i.e., inhaled corticosteroids, salmeterol, cromolyn, and theophylline) that were utilized, as recommended, by the National Asthma Expert Panel (NAEP) of the National Heart, Lung, and Blood Institute (NHLBI) guidelines were 70 by AIs vs. 24 by PCPs over three visits. Cromolyn was prescribed five times over three visits by AIs and not at all by PCPs. Recognition and treatment of coexisting allergic rhinitis was evident in only 13% of patients treated by PCPs as compared to 80% in those treated by AIS. (p < 0.0001). However, all patients treated by AIs were skin tested to explore the presence of allergic triggers, while no patients treated by PCPs were evaluated for IgE-mediated reactions. Treatment cost for allergic rhinitis was therefore higher, at $2039, for AIs as compared to $741 for PCPs. There were no peakflow values in charts obtained from PCPs. However, all charts from AIs had peakflow values, which improved during the course of therapy in 33% of patients. Total medication costs for asthma were higher for AIs @ $5,646.30 vs. $1,932.25 for PCPs. Total medication costs for allergic rhinitis plus asthma were higher for AIs @ $7615 vs. $2681 for PCPs. However, patients treated by AIs had more severe asthma and required more frequent visits. Ipratropium bromide was prescribed a total of four times over several visits by PCPs vs. only once by AIs. In comparing asthma care between AI specialists and PCPs, it was found that AI specialists treat more severe asthmatics, provide more frequent follow-up visits, utilize peak flow rates, prescribe more controller medications, and more often recognize and treat comorbid conditions such as allergic rhinitis that impact on asthma care. Thus, although treatment costs for AIs are higher, these costs are justified by a quality of care that is more consistent with national (NHLBI) guidelines.

Adult↗

Axonal injury in falls.

Amyloid precursor protein (APP) immunocytochemistry was used as a marker for axonal injury (AI) in a series of 16 cases of head trauma associated with fatal falls. Nine cases were falls from not more than the person's own height (falls from < or = own height) and seven cases were falls from a distance greater than the person's own height (falls from > own height). AI was recorded on a series of line diagrams of standard brain sections divided into 116 sectors. AI around focal lesions (infarcts, hemorrhages, contusions) was distinguished from nonfocal axonal injury that was distant from any focal area of damage. The percentage of sectors showing focal AI provided the Focal Axonal Injury Score (FAIS) and the percentage showing nonfocal AI the Non-Focal Axonal Injury Score (NFAIS). The FAIS is a measure of secondary AI and the NFAIS of diffuse axonal injury (DAI). The percentage of sectors involved with AI (focal and nonfocal) provided the cumulative Axonal Injury Score (AIS). A semiquantitative grading system was also used to assess the severity of axonal injury in each sector and the sum of the grades from all sectors was expressed as a percentage to provide the Axonal Injury Severity Score (AISS). Widespread AI was present in all cases irrespective of the height of the fall. AI was present in the midbrain (94%), pons (94%), corpus callosum (100%), central grey matter (100%), and cerebral hemispheric white matter (94%). AIS ranged from 10 to 94 in falls from < or = own height (mean 73) and from 38 to 92 in falls from > own height (mean 82). AISS ranged from 6 to 95 in falls from < or = own height (mean 65) and 28 to 95 in falls from > own height (mean 72). There was no statistically significant difference in AIS or AISS between the two groups. The extent and severity of AI cannot be predicted from biomechanical data, such as the height of the fall, as the total AI in a given case is a variable mixture of Nonfocal AI (DAI) and Focal AI arising by secondary mechanisms, and APP immunostaining is unable to distinguish primary from secondary AI. However, the combination of the Hypoxic-Ischemic Score (HIS) defined as the percentage of sectors showing any hypoxic-ischemic damage ranging from neuronal "red cell change" to infarction in conjunction with the FAIS and NFAIS provided a measure of the relative contribution of primary and secondary AI in a given brain.

Accidental Falls↗

Accommodative insufficiency is the primary source of symptoms in children diagnosed with convergence insufficiency.

PURPOSE: Accommodative insufficiency (AI) and convergence insufficiency (CI) have been associated with similar symptomology and frequently present at the same time. The severity of symptomology in CI has been linked to the severity of the CI, suggesting a dose-dependent relationship. However, with increasing severity of CI also comes increased comorbidity of AI. AI alone has been shown to cause significant symptomology. We hypothesize that AI drives the symptoms in CI with a comorbid AI condition (CIwAI) and that it is the increased coincidence of AI, rather than increased severity of CI, which causes additional symptomology. METHODS: Elementary school children (n = 299) participated in a vision screening that included tests for CI and AI and the CISS-V15 symptom survey. They were categorized into four groups:1) normal binocular vision (NBV); 2) AI-only; 3) CI-only; and 4) CIwAI. One hundred seventy elementary school children fell into the categories of interest. RESULTS: Pairwise comparison of the group means on the symptom survey showed: 1) children with AI-only (mean = 19.7, p = 0.006) and children with CIwAI (mean = 22.8, p = 0.001) had significantly higher symptom scores than children with NBV (mean = 10.3); and 2) children with CI-only (mean = 12.9, p = 0.54) had a similar symptom score to children with NBV. Using a two-factor analysis of variance (AI and CI), the AI effect was significant (AI mean = 21.56; no AI mean = 11.56, p < 0.001), whereas neither the CI effect (p = 0.16) nor the CI by AI interaction effect (p = 0.66) were significant. CONCLUSION: CI is a separate and unique clinical condition and can occur without a comorbid AI condition, our CI-only group. Past reports of high symptom scores for children with CI are the result of the presence of AI, a common comorbid condition. When AI is factored out, and children with CI only are evaluated, they are not significantly more symptomatic than children with NBV.

Accommodation, Ocular↗

Modulation of luminescence operon expression by N-octanoyl-L-homoserine lactone in ainS mutants of Vibrio fischeri.

Population density-dependent expression of luminescence in Vibrio fischeri is controlled by the autoinducer N-3-oxohexanoyl-L-homoserine lactone (autoinducer 1 [AI-1]), which via LuxR activates transcription of the lux operon (luxICDABEG, encoding the putative autoinducer synthase [LuxI] and the luminescence enzymes). We recently identified a novel V. fischeri locus, ainS, necessary for the synthesis of a second autoinducer, N-octanoyl-L-homoserine lactone (AI-2), which via LuxR can activate lux operon transcription in the absence of AI-1. To define the regulatory role of AI-2, a luxI ainS double mutant was constructed; in contrast to the parental strain and a luxI mutant, the luxI ainS mutant exhibited no induction of luminescence and produced no detectable luminescence autoinducer, demonstrating that V. fischeri makes no luminescence autoinducers other than those whose synthesis is directed by luxI and ainS. A mutant defective only in ainS exhibited accelerated luminescence induction compared with that of the parental strain, indicating that AI-2 functions in V. fischeri to delay luminescence induction. Consistent with that observation, the exogenous addition of AI-2 inhibited induction in a dose-dependent manner in V. fischeri and Escherichia coli carrying the lux genes. AI-2 did not mediate luxR negative autoregulation, alone or in the presence of AI-1, and inhibited luminescence induction in E. coli regardless of whether luxR was under the control of its native promoter or a foreign one. Increasing amounts of AI-1 overcame the inhibitory effect of AI-2, and equal activation of luminescence required 25- to 45-fold-more AI-2 than AI-1. We conclude that AI-2 inhibits lux operon transcription. The data are consistent with a model in which AI-2 competitively inhibits the association of AI-1 with LuxR, forming a complex with LuxR which has a markedly lower lux operon-inducing specific activity than that of AI-1-LuxR. AI-2 apparently functions in V. fischeri to suppress or delay induction at low and intermediate population densities.

Bacterial Proteins↗