Effect of Removing Seminal Plasma Using a Sperm Filter on the Viability of Refrigerated Stallion Semen

Cooling of equine semen obtained from some stallions results in lower seminal quality and viability when the seminal plasma (SP) is present. The objective of this study was to evaluate the effect of the removal of SP using a Sperm Filter on the viability of cooled stallion semen. For this purpose, 31 stallions were used. Their ejaculates were divided into three groups: CN, semen was diluted with an extender; FLT, SP was removed by filtration; and CT, SP was removed by centrifugation and cooled to 15°C for 24 hours. Sperm kinetics and plasma membrane integrity were evaluated immediately after collection (T0) and after 24 hours of refrigeration (T1). No difference (P > .05) was noted at T1 for total sperm motility (TM), progressive sperm motility, or plasma membrane integrity when semen samples from all the stallions were analyzed. However, when samples from stallions termed bad coolers were analyzed (TM = <30% at T1), a difference was observed in TM and progressive sperm motility for CN compared with FLT and CT at T1. Sperm recovery was greater when SP was removed using the filter (FLT) to that when the SP was removed by centrifugation (CN) (89% vs. 81%). Thus, we concluded that filtering with a Sperm Filter is an efficient and practical method for removal of SP from stallion ejaculates, with lower sperm loss than centrifugation. We also found that the presence of SP reduces the quality and viability of cooled semen from stallions whose semen is sensitive to the process of refrigeration. © 2013 Elsevier Inc.

Graft incorporation and implant osseointegration following the use of autologous and fresh-frozen allogeneic block bone grafts for lateral ridge augmentation

Objectives: To compare autogenous bone (AT) and fresh-frozen allogeneic bone (AL) in terms of histomorphometrical graft incorporation and implant osseointegration after grafting for lateral ridge augmentation in humans. Materials and methods: Thirty-four patients were treated with either AL (20 patients) or AT (14 patients) onlay grafts. During implant installation surgery 6 months after grafting, cylindrical biopsies were harvested perpendicularly to the lateral aspect of the augmented alveolar ridge. Additionally, titanium mini-implants were installed in the grafted regions, also perpendicularly to the ridge; these were biopsied during second-stage surgery. Histological/histomorphometric analysis was performed using decalcified and non-decalcified sections. Results: Histological analysis revealed areas of necrotic bone (NcB) occasionally in contact with or completely engulfed by newly formed vital bone (VB) in both AT and AL groups (55.9 ± 27.6 vs. 43.1 ± 20.3, respectively; P = 0.19). Statistically significant larger amounts of VB (27.6 ± 17.5 vs. 8.4 ± 4.9, respectively; P = 0.0002) and less soft connective tissue (ST) (16.4 ± 15.6 vs. 48.4 ± 18.1, respectively; P ≤ 0.0001) were seen for AT compared with AL. No significant differences were observed between the groups regarding both bone-to-implant contact (BIC) and the bone area between implant threads (BA) on the mini-implant biopsies. Conclusion: Allogeneic bone block grafts may be an option in cases where a limited amount of augmentation is needed, and the future implant can be expected confined within the inner aspect of the bone block. However, the clinical impact of the relatively poor graft incorporation on the long-term performance of oral implants placed in AL grafts remains obscure. © 2013 John Wiley & Sons A/S.

Identification of sperm head subpopulations with defined pleiomorphic characteristics in ejaculates of captive Goeldi's monkeys (Callimico goeldii)

The aim of this study was to evaluate the incidence of pleiomorphisms and its influence on the distribution of sperm morphometric subpopulations in ejaculates from the vulnerable Goeldi's monkey (Callimico goeldii) by using a combination of computerized analysis system and Principal Component Analysis (PCA) methods Each sperm head was measured for four primary spermatozoal head dimensional parameters (area [A (μm2)], perimeter [P (μm)], length [L (μm)] and width [W (μm)]) and three head shape derived parameters (ellipticity [(L/W)], elongation [(L-W)/(L+W)] and rugosity [(4πA/P2)]) Six separate subpopulations (SPs) were identified: SP1, constituted by very large, narrow and very elliptical spermatozoa (A=16.85±1.56μm2, W=2.75±0.42μm and ellipticity=2.16±0.24); SP2, characterized by average sized, short, wide and round spermatozoa (A=15.00±1.92μm2, L=5.06±0.49μm, W=3.51±0.31μm and ellipticity=1.44±0.15); SP3, represented by small, wide and slightly round spermatozoa (A=14.95±1.75μm2, W=3.47±0.29μm and ellipticity=1.48±0.14); SP4 included very small, short and very round spermatozoa (A=14.15±2.38μm2, L=4.90±0.57μm and elongation=0.18±0.05); SP5 consisted of average sized and slightly elliptical spermatozoa (A=15.14±1.72μm2 and ellipticity=1.49±0.14); and SP6 included large and round spermatozoa (A=16.30±1.62μm2 and elongation=0.19±0.04) There were differences in the sperm subpopulation distribution (P<0.001) among the five donors analyzed In conclusion, the results of the current study confirmed that the use of computer sperm analysis methods combined with PCA cluster analyses are useful methods to identify, classify, and characterize different sperm head morphometric subpopulations in neotropical primates Broadening our knowledge of C goeldii sperm morphometric abnormalities as well as developing reliable techniques for sperm evaluation may be essential for ex situ conservation of this threatened species © 2012 Elsevier B.V.

Fate of autologous and fresh-frozen allogeneic block bone grafts used for ridge augmentation. A CBCT-based analysis

Objectives: To evaluate dimensional changes in autologous (AT) and fresh-frozen allogeneic (AL) block bone grafts 6 months after alveolar ridge augmentation. Material and methods: Twenty-six partially or totally edentulous patients treated either with fresh-frozen AL bone or AT bone onlay block grafts prior to implant placement (13 patients in each group), were included in this analysis. Patients received CBCT (i-CAT Classic) examinations prior to surgery and 14 days and 6 months after grafting. Differences in alveolar ridge area among the various observation times were evaluated by planimetric measurements on two-dimensional CBCT images of the grafted regions. Nineteen grafted blocks from each group were evaluated. Results: Significant increase in alveolar ridge dimensions, allowing implant placement, was obtained with both types of grafts 6 months after grafting; no significant differences in alveolar ridge area were observed between the groups at the various observation times. However, graft resorption in the AL group was significantly larger compared to that in the AT group at 6 months. Conclusions: Larger bone graft resorption was seen in patients treated with fresh-frozen AL bone than in those treated with AT bone 6 months following alveolar ridge augmentation. © 2011 John Wiley & Sons A/S.

Effect of Storage Time and Temperature of Equine Epididymis on the Viability, Motion Parameters, and Freezability of Epididymal Sperm

The recovery of sperm from the epididymal cauda may be the last chance to obtain genetic material when sudden death or serious injuries occur in valuable stallions. However, the lack of technical knowledge regarding the storage and transportation of the epididymis often prevents the preservation of the sperm. Therefore, the aim of this study was to compare sperm parameters of sperm obtained immediately after orchiectomy with sperm recovered from epididymal cauda at different times after storage at 5°C and at room temperature (RT). For that, 48 stallions of different breeds were used. In group 1 (control group), eight stallions were used, and the harvest of the epididymal sperm was performed immediately after orchiectomy. In group 2, 40 stallions were used, which were divided into five groups according to the storage time of the epididymis after orchiectomy (6, 12, 18, 24, or 30 hours), making a total of eight stallions per group. One epididymis of each stallion was stored at 5°C, and the contralateral epididymis was stored at RT, both for the same period. The sperm parameters of total motility, progressive motility, progressive linear velocity, curvilinear velocity, percentage of rapid sperm, and plasma membrane integrity were evaluated in all the groups after sperm recovery, resuspension in a sperm freezing diluent, and thawing. In conclusion, the storage of the testis-epididymis complex at 5°C provided better preservation of epididymal sperm than the storage at RT, and regardless of the temperature, the progressive motility is the sperm parameter that is most sensitive to storage time. © 2013 Elsevier Inc.

High incidence of 'Dag-like' sperm defect in the domestic cat

The occurrence of a high incidence of sperm tail defects in a male domestic cat resembling the known 'Dag-like' defect is reported. Sperm analyses were performed in ejaculated samples collected by an artificial vagina and in testicular and epididymal sperm cells after castration. The following alterations were observed using transmission electron microscope: heavily coiled sperm tails containing several axonemal units enclosed in the same common cell membrane; aberrations in the axonemal main structure; and swollen and unevenly distributed mitochondria in the midpiece. Abnormal modifications in the mitochondrial sheath were also found in sperm cells retrieved from testes and epididymides. Considering these findings, we can conclude that this is the Dag-like defect, described previously in other domestic species and a testicular origin may be involved. © ISFM and AAFP 2012.

Is peripheral blood cell balanced altered by the use of fresh frozen bone block allografts in lateral maxillary ridge augmentation?

Background: The relationship between the immune response and red and white blood cell homeostasis is cited in literature, but no studies regarding the balance of these cell populations following maxillary bone-graft surgeries can be found. Aim: The aim of this study was to evaluate the possible impairments in the blood cell balance following fresh-frozen allogeneic bone-graft augmentation procedures in patients who needed maxillary reconstruction prior to implants. Material and Methods: From 33 patients elected to onlay bone grafting procedures, 20 were treated with fresh-frozen bone allografts and 13 with autologous bone grafts. Five blood samples were collected from each patient in a 6-month period (baseline: 14, 30, 90, and 180 days postsurgery), and the hematological parameters (erythrogram, leukogram, and platelets count) were accessed. Results: All evaluated parameters were within the reference values accepted as normal, and significant differences were found for the eosinophils count when comparing the treatments (30 days, p=.035) and when comparing different periods of evaluation (allograft-treated group, baseline×180 days, p≤.05 and 90×180 days, p≤.01; autograft-treated group, 30×90 days, p≤.05 and 30×180 days, p≤.05). Conclusions: Both autologous and fresh-frozen allogeneic bone grafts did not cause any impairment in the red and white blood cell balance, based on quantitative hemogram analysis, in patients subjected to maxillary reconstruction. © 2011 Wiley Periodicals, Inc.

Changes during chilled storage of whole tilapia and short-term frozen storage of tilapia fillets

This study evaluated the physicochemical changes in Nile tilapia (n = 82, 373.71 ± 61.91 g) refrigerated for up to 92 h and in the frozen fillets. The tilapias were captured with nets, slaughtered by ice and water shock (1:1) in a temperature of approximately 2°C for 30 min, and stored refrigerated at 4°C in polystyrene boxes containing ice. The fish were filleted, and filets were weighed and frozen. The drip loss and protein were determined after 23 days of frozen storage. After 4 h of storage, all fish were in full rigor mortis. The pH of the muscles decreased for up to 45 h of the storage period. The fillets obtained from tilapia stored for more than 72 h lost more weight and protein. Thus, the filleting or processing of tilapia should be done before 72 h of cold storage, since deterioration of the fish starts to occur after this period. Copyright © Taylor & Francis Group, LLC.

Healing of fresh frozen bone allograft with or without platelet-rich plasma: A histologic and histometric study in rats

Purpose: This study histomorphometrically analyzed the effect of autogenous platelet-rich plasma (PRP) on healing of fresh frozen bone allograft (FFBA) in bony defects in rat calvaria. Materials and Methods: A 5mm-diameter defect was created in the calvarium of 30 rats. Animals were divided into three groups: C (defect was filled by blood clot only), FFBA (defect was filled with 0.01mL of FFBA), and FFBA/PRP (defect was filled with 0.01mL of FFBA combined with 100μL of PRP). All animals were euthanized at 30 days postoperatively. Histomorphometry and histology analyses were performed. Data were statistically analyzed (analysis of variance, Tukey, p<.05). Results: FFBA had a statistically smaller new bone area than groups FFBA/PRP and C. No statistically significant differences were observed between groups FFBA and FFBA/PRP with regard to remaining bone graft particle area. Conclusion: It can be concluded that (1) PRP improved the incorporation of FFBA, increasing the amount of new bone formed; (2) PRP has not influenced the resorption of nonviable particles of the FFBA; and (3) presence of remaining FFBA particles might have accounted for the smaller amount of new bone observed in group FFBA when compared with control group. © 2011 Wiley Periodicals, Inc.

Slimmer or Fertile? Pharmacological Mechanisms Involved in Reduced Sperm Quality and Fertility in Rats Exposed to the Anorexigen Sibutramine

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of Pentoxifylline on Equine Epididymal Sperm

This study evaluated whether pentoxifylline (PTX) present in the flushing extender influenced the function of equine epididymal spermatozoa after recovery and after thawing. For this experiment, 58 testicles from 29 Brazilian Jumping Horses were used. Cauda epididymides of each stallion were separated and flushed with a skim milk extender, with or without 7.18 mM PTX and then subjected to the freezing process. Samples flushed with the extender containing PTX showed a significant increase in total motility, progressive motility, straight line velocity, curvilinear velocity, and percentage of rapid sperm immediately after the recovery of epididymal sperm and after 15 minutes of incubation at 37°C (P < .05). However, the presence of PTX in the flushing extender did not affect the post-thaw motility parameters or plasma membrane integrity (P > .05). The results of this study showed that the PTX present in the flushing extender improved motility parameters of recently recovered epididymal sperm and had no deleterious effects on plasma membrane integrity and freezability of equine epididymal sperm. © 2013 Elsevier Inc. All rights reserved.

Clinical similarities and histological diversity comparing fresh frozen onlay bone blocks allografts and autografts in human maxillary reconstruction

Background: In the absence of autologous bone for harvesting, fresh-frozen bone allografts turned into an alternative for bone reconstruction procedures. Purpose: The purpose of this study was to make a histological analysis of fresh-frozen onlay bone allografts (ALs), compared with autografts, in patients who needed maxillary reconstruction prior to dental implants placement. Materials and Methods: Twelve patients with bone deficiencies (width inferior to 4mm) in the sites where the implants were planned were enrolled in the study. From these, six were elected to be treated with autogenous (AT) bone grafts and six with fresh-frozen bone AL. This last group included the patients who had absence of a convenient amount of bone in donor sites. Each patient received from one to six graft blocks, totalling to 12 ATs and 17 ALs. Seven months after grafting procedures, biopsies of the grafts were made using 2-mm internal diameter trephine burs, and processed for histological analysis. One biopsy was retrieved from each patient. Results: Clinically, all grafts were found to be firm in consistency and well-incorporated to the receptor bed. Histological analysis showed a large amount of necrotic bone surrounded by few spots of new-formed bone in the AL group, suggesting low rate of graft remodeling. In the AT group, an advanced stage of bone remodeling was seen. Conclusions: Human fresh-frozen bone block AL showed clinical compatibility for grafting procedures, although associated to slow remodeling process. Further studies are needed to define, at long term, the remodeling process chronology the clinical longitudinal results for fresh-frozen bone AL. Copyright © 2013 Wiley Periodicals, Inc.

Immunological aspects of fresh-frozen allogeneic bone grafting for lateral ridge augmentation

Objectives: To present some immunological aspects of fresh-frozen allogeneic bone grafting for lateral bone augmentation, based on the quantitative evaluation of IL-10, IL-1β, IFN- γ and TNF- α in patients sera. Material and methods: Thirty-three partially or totally edentulous patients received fresh-frozen allogeneic bone (AL - 20 patients) or autologous bone onlay block grafts (AT - 13 patients) prior to oral implant placement. Blood samples were collected from each patient at various time-points during a 6 month-period (baseline, 14, 30, 90 and 180 days postoperatively). Quantitative evaluation of IL-10, IL-1β, IFN- γ and TNF- α was performed by enzyme linked immunosorbent assay (ELISA). Results: For all evaluated markers and at all evaluated periods, inter-group comparisons showed no statistically significant differences between the groups, while the observed values were within normal levels. For AL-treated patients, intra-group evaluation showed statistically significant increase of TNF-α from baseline to 90 (P < 0.001) and 180 (P < 0.01) days, and from 14 to 90 (P < 0.01) and 180 (P < 0.05) days. IFN- γ showed intercalated results, with a decrease from baseline to 14 days (P < 0.05), and increase from 14 to 90 days (P < 0.001) and 180 (P < 0.05) days. No differences between the periods of evaluation were found for the AT group. Conclusions: AL grafting for lateral bone augmentation, similar to AT grafting, does not seem to challenge the immune system significantly. © 2012 John Wiley & Sons A/S.

Cooling of ejaculated and epididymal stallion sperm

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

The relationships between scrotal surface temperature, age and sperm quality in stallions

In horses, spermatogenesis normally occurs at an average intratesticular temperature of 35. °C; therefore, mechanisms for testicular thermoregulation are essential. Measuring the scrotal surface temperature by thermography is one of the methodologies used to evaluate the effectiveness of testicular thermoregulation. The objective of this study was to determine the relationship between the control of scrotal surface temperature and sperm quality in horses of different ages. In total, 24 Quarter Horse stallions were divided into three groups: YS (young stallions), AS (adult stallions) and OS (old stallions). Initially, we calculated the testicular volume (TV) and evaluated various aspects of the semen (sperm kinetics, plasma membrane integrity and sperm morphology) for all the animals. We also evaluated rectal temperature (RT), body surface temperature (BST,) and average scrotal surface temperature in the testicular region (SST) before (M0) and after sun exposure (M1). Differences were observed (p<0.05) between the RT and BST before and after sun exposure in all three groups. However, there were no differences (p>0.05) in the SST values at these two time points, thus demonstrating the efficiency of the mechanisms for testicular thermoregulation. The SST was similar (p>0.05) among all three groups. Based on these results, we conclude that fertile stallions of different age groups are able to maintain SST and measuring the heat radiating from the scrotum using a digital infrared thermographer. We can also conclude that measuring the heat radiating from the scrotum using a digital infrared thermographer is a practical and efficient tool for monitoring SST in horses. © 2013 Elsevier B.V.