The effect of platelet-rich plasma (PRP) combined with a bone allograft on human periodontal ligament (PDL) cells

The prominent purpose of the study was the evaluation of the in vitro mitogenic effect of three different homologous platelet-rich plasma (PRP) preparations (PRPa, PRPb, PRPc) on three different lines of periodontal ligament (PDL) cells (PDL_1,2,3), cultured alone or in combination with a demineralized freeze-dried allograft (DFBA). PDL cell cultures were derived from the mid root of three maxillary caries-free premolars extracted for orthodontic reasons. Cells were grown and reached confluence. To evaluate the mitogenic effect of all exogenous factors (PRPa, PRPb, PRPc and DFBA) on PDL cells, specific number of cells (10.000/well) was cultured in the presence or absence of the above factors. Each PRP preparation (5% v/v) was added in all cell lines, in the absence or presence of 10 mg/ml of DFBA. The cells were also treated with 25 ng/ml bFGF (positive control). The mitogenic effect was evaluated 24 h after incubation, using the Trypan blue exclusion assay. The results revealed that all PRP preparations act as potent mitogens as they significantly induced cell proliferation on PDL_1,2,3 lines. All PRP preparations when added alone in the PDL cell cultures, exhibited a significant advantage over the positive control (bFGF). The addition of DFBA to PRP did not influence significantly cell proliferation in all cell lines, comparatively to PRP alone, at the time -period studied. The findings of this study demonstrate the beneficial role of PRP alone or combined with the bone graft on periodontal ligament cells in vitro, suggesting that it may be considered as a potential biological approach in periodontal regeneration.     

A novel collagen/platelet-rich plasma (COL/PRP) scaffold: preparation and growth factor release analysis

Platelet-rich plasma (PRP) has been widely used in clinical practice for more than 20 years because it causes the release of many growth factors. However, the burst release pattern and short release period of PRP have become obstacles to its application. An optimal controllable release system is an urgent need for researchers. This study investigated whether collagen/PRP (COL/PRP) scaffolds can serve as a vehicle for the controllable release of growth factors. We fabricated a novel scaffold that integrates PRP activated by thrombin or collagen into type I collagen. The mechanical properties, cytotoxicity, and transforming growth factor β1 (TGF-β1), platelet derived growth factor (PDGF), fibroblast growth factor (FGF) and vascular endothelial growth factor (VEGF) content were evaluated. Our results demonstrate that the COL/PRP scaffolds were not cytotoxic to L-929 fibroblasts. The PDGF and FGF content in the thrombin group was at a higher level and lasted for a long period of time. Collagen and thrombin played the same role in the release of TGF-β1 and VEGF. These data suggest that the novel COL/PRP scaffolds provide a carrier for the controllable release of growth factors and may be used in tissue- regenerative therapies.     

A radioimmunoassay for total (unconjugated + conjugated) oestrone in plasma from pregnant women; the response of total oestrone in plasma to an intravenous injection of dehydroepiandrosterone sulphate (DHAS).

A non-chromatographic radioimmunoassay for estimation of total oestrone in plasma from pregnant women is described. A purified Helix pomatia extract is used for the hydrolysis. The antiserum has a high specificity to oestrone. The technical procedure is simple and rapid. Only small amounts of plasma (0.01–0.05) are needed for the analysis. The method has been applied to the measurement of total oestrone in plasma from pregnant women. During normal pregnancy a gradual increase of total oestrone was noticed from about 75 ng/ml in the 33rd–34th week to slightly more than 100ng/ml in the 39th-40th week. Great individual variations were observed. Patients giving birth to small-for-date children showed a tendency towards lower values. In response to an intravenous injection of dehydro-epiandrosterone sulphate (DHAS) a marked increase of total oestrone in plasma was recorded with peak values 360min after the injection. A considerable individual variation in the magnitude of the rise was observed. The rise was generally lower in patients with small-for-date babies. The results do not suggest that measurement of total oestrone before or after an injection of DHAS will be useful for assessment of placental function.     

Circulating plasma alpha-tocopherol following a single injection in a black rhinoceros (Diceros bicornis)

Injectable all rac-alpha-tocopherol, at a dose of 12.4 IU/kg body mass, increased circulating levels of alpha-tocopherol in the black rhinoceros (Bicornis diceros) from 0.18 microgram/ml to 1.47 micrograms/ml within 2 hr. Although the plasma level peaked at Day one (13.07 micrograms/ml) and dropped rapidly, substantial residual effects were seen even 10 days (1.50 micrograms/ml) following a single injection. It appears that parenteral vitamin E administration may be suitable for therapeutic treatment of vitamin E deficiency in the black rhinoceros.     

Surface ultrastructure of uterine epithelial cells in women with premature ovarian failure following steroid hormone replacement

Scanning electron microscopy has been used to study the apical surface of uterine epithelial cells in women with premature ovarian failure following steroid hormone replacement therapy. A variety of ultrastructural characteristics are identified which could indicate a uterus that is receptive for blastocyst implantation.     

A Comparison between Platelet-Rich Plasma (PRP) and Hyaluronate Acid on the Healing of Cartilage Defects

Platelet-rich plasma (PRP) has offered great promise for the treatment of cartilage degradation, and has been proved to have positive effects on the restoration of cartilage lesions. But no comparative work has been done between PRP and hyaluronate acid (HA) concerning their restoring effect on cartilage defect, especially by means of animal experiments and histologic assessments. The purpose of the study was to compare the therapeutic effects of P-PRP and HA on osteoarthritis in rabbit knees. Thirty rabbits were used to establish the animal models by creating a cartilage defect of 5 mm in diameter on the condyles of the femurs, and were randomly divided into three groups: the P-PRP group, HA group and the control group. Then each group was treated with P-PRP, HA or saline solution, respectively. Six and twelve weeks later the rabbits were sacrificed and the samples were collected. The platelet number, the concentrations of growth factors of P-PRP and whole blood, and the IL-1β concentration in the joint fluid were investigated, and the histological assessment of the cartilage were performed according to Mankin''s scoring system. Micro-CT was also used to evaluate the restoration of subchondral bone. The platelet concentration in P-PRP is 6.8 fold of that in the whole blood. The IL-1β level in the P-PRP group was lower than in the HA group (p<0.01) and in the control group (p<0.01). The restoration of the defected cartilage as well as the subchondral bone was better in the P-PRP group than in the HA group or the control group (P<0.05). Our data showed that P-PRP is better than HA in promoting the restoration of the cartilage and alleviating the arthritis caused by cartilage damage.     

Effects of an analog of PGF2(alpha) (ONO-1052) on cows with luteinized ovarian cysts following treatment with GnRH analog (fertirelin acetate)

Effects of PGF2(alpha) analog (ONO-1052) on cows with luteinized cysts following treatment with GnRH analog (fertirelin) in inducing estrus and shortening an interval from treatment to conception were investigated in a field trial. Seventy-five cows with follicular cysts diagnosed by rectal palpation were treated intramuscularly (i.m.) with 400 mug fertirelin (an analog of GnRH). Luteinization of follicular cysts were tentatively judged by rectal palpation. Forty-one cows were considered to have luteinized cysts and were either treated i.m. with ONO-1052 (28 cows) or left untreated as controls (13 cows). Thirty-four other cows were considered to have not responded to fertirelin and retreated with 10,000 MU hCG (19 cows) or fertirelin (five cows). This tentative judgment of luteinization of the cysts by rectal palpation after treatment was later confirmed by determining serum progesterone concentration before and 10 to 14 days after treatment. Only in the cows with luteinized cysts which were confirmed by serum progesterone analysis; increased from a pretreatment level below 1.0 ng/ml to a value of 1.0 ng/ml or higher 10 to 14 days after treatment, effects of ONO-1052 combined with fertirelin were investigated. Of the 18 cows with luteinized cysts thus confirmed following fertirelin injection and treated with ONO-1052, 15 (83.3 %) cows came into estrus within 26 +/- 14 (mean +/- SD) days after the first treatment; eight cows within three to five days and four cows within 22 to 28 days, and 14 cows (77.8 %) conceived within 100 days (42 +/- 26 days). On the other hand, the five control cows with luteinized cysts that were not treated with ONO-1052 required significantly longer to exhibit normal estrus (54 +/- 13 days; P<0.05) and to conceive (54 +/- 13 days). These results indicate that ONO-1052 combined with fertirelin may be useful to shorten the interval from treatment to normal estrus and to conception in cows with follicular cysts.     

Oral Ezatiostat HCl (TLK199) and Myelodysplastic syndrome: A case report of sustained hematologic response following an abbreviated exposure

Treatment options for patients with lower risk non-del(5q) myelodysplastic syndromes (MDS) who fail erythroid stimulating agents are restricted to one of the hypomethylating drugs with an expected response rate of ~50%. Ezatiostat HCl, an agent with the potential for producing multi-lineage responses in this population is currently in clinical investigation phase. This case report describes a 77 year old male who received less than two cycles of therapy with ezatiostat HCl which had to be aborted due to intolerable side effects, but which produced a sustained normalization of all three blood counts. This trilineage response has now lasted for more than a year. Interestingly, the patient began with a del(5q) abnormality and responded briefly to lenalidomide. Upon relapse of the anemia, a bone marrow showed the disappearance of the del(5q) but the appearance of a new clonal abnormality t(2;3). Given that the patient had a complete cytogenetic response to a truncated exposure to lenalidomide followed by a trilineage response to an even briefer course of ezatiostat HCl suggests a potential role for ezatiostat HCl in del(5q) patients who relapse following lenalidomide.     

Mannose-Binding Lectin (MBL) and MBL-associated serine protease-2 (MASP-2) in women with malignant and benign ovarian tumours

Mannose-Binding Lectin (MBL) is a serum pattern recognition molecule, able to activate complement in association with MASP proteases. Serum levels of MBL and MASP-2, activities of MBL–MASP complexes, single nucleotide polymorphisms of the MBL2 and MASP2 genes and/or their specific mRNA expression in ovarian sections were investigated in 128 patients suffering from primary ovarian cancer (OC) and compared with 197 controls (C), encompassing both patients with benign ovarian tumours (n = 123) and others with no ovarian pathology (n = 74). MBL deficiency-associated genotypes were more common among OC patients than among controls. The O/O group of genotypes was associated with ovarian cancer (OR 3.5, p = 0.02). In A/A homozygotes, MBL concentrations and activities were elevated in the OC group and correlated with C-reactive protein. Moreover, high MBL serum levels were associated with more advanced disease stage. No differences in distribution of the MASP2 +359 A>G (D120G) SNP or MASP-2 serum levels were found between cancer patients and their controls. However, the highest frequency of the A/G (MASP2) and LXA/O or O/O (MBL2) genotypes was found among OC patients with tumours of G1–2 grade (well/moderately differentiated). Furthermore, MBL deficiency-associated genotypes predicted prolonged survival. None of the parameters investigated correlated with CA125 antigen or patients’ age. The local expression of MBL2 and MASP2 genes was higher in women with ovarian cancer compared with controls. It is concluded that the expression of MBL and MASP-2 is altered in ovarian cancer, possibly indicating involvement of the lectin pathway of complement activation in the disease.     

Pregnancy outcomes in women with repeated implantation failures after intracytoplasmic morphologically selected sperm injection (IMSI)

Background

The role of serum anti-Müllerian hormone (AMH) as predictor of in-vitro fertilization outcomes has been much debated. The aim of the present study is to investigate the practicability of combining serum AMH level with biological age as a simple screening method for counseling IVF candidates of advanced reproductive age with potential poor outcomes prior to treatment initiation.

Methods

A total of 1,538 reference patients and 116 infertile patients aged greater than or equal to 40 years enrolled in IVF/ICSI cycles were recruited in this retrospective analysis. A reference chart of the age-related distribution of serum AMH level for Asian population was first created. IVF/ICSI patients aged greater than or equal to 40 years were then divided into three groups according to the low, middle and high tertiles the serum AMH tertiles derived from the reference population of matching age. The cycle outcomes were analyzed and compared among each individual group.

Results

For reference subjects aged greater than or equal to 40 years, the serum AMH of the low, middle and high tertiles were equal or lesser than 0.48, 0.49-1.22 and equal or greater than 1.23 ng/mL respectively. IVF/ICSI patients aged greater than or equal to 40 years with AMH levels in the low tertile had the highest cycle cancellation rate (47.6%) with zero clinical pregnancy. The nadir AMH level that has achieved live birth was 0.56 ng/mL, which was equivalent to the 36.4th percentile of AMH level from the age-matched reference group. The optimum cut-off levels of AMH for the prediction of nonpregnancy and cycle cancellation were 1.05 and 0.68 ng/mL, respectively.

Conclusions

Two criteria: (1) age greater than or equal to 40 years and (2) serum AMH level in the lowest tertile (equal or lesser than 33.3rd percentile) of the matching age group, may be used as markers of futility for counseling IVF/ICSI candidates.     

Decrease of plasma somatotropin levels following thyroliberin (TRH) administration in patients with hyperthyroidism.

Thyroliberin was observed to induce a significant decrease of plasma somatotropin levels in patients with untreated hyperthyroidism, except those with very low initial somatotropin levels. These data have confirmed our previous finding where we noted that under the conditions of thyroid hormone excess thyroliberin was able to exert an inhibitory effect on somatotropin release.     

First report on antibody response of Seriola dumerilii (Risso 1810) challenged with Listonella anguillarum

The greater amberjack, Seriola dumerilii (Risso 1810) is a semi-pelagic fish and a worldwide species; it is considered a promising candidate for the diversification of Mediterranean aquaculture. In this paper an experimental injection with Listonella (Vibrio) anguillarum was performed to study the immune response of S. dumerilii. Antibody titres to L. anguillarum O1 were determined with indirect-ELISA at different times over a period of 42 days. Results showed that the antibody levels against L. anguillarum were significantly higher in the challenged fish compared to the control. They started developing since the 5th day reaching the highest peak on day 20 after injection, indicating a fast response of the immune system. The observed antibody titre was very high versus L. anguillarum if compared to other fish species.     

A clinical trial of live attenuated varicella vaccine (Biken) in children with malignant diseases

The live attenuated varicella vaccine (Biken) derived from the Oka strain was used for immunization of childhood cancer patients in remission but on chemotherapy. Thirty nine patients were immunized without any severe adverse effects but 6 recipients had a small number of vesicles. Seroresponses were observed in 90% of recipients examined. During the follow-up period 6 leukemia recipients were suffered from natural varicella in family or community contacts. No vaccinee developed herpes-zoster during this study period.