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1.
In Vitro Cell Dev Biol Anim ; 56(10): 859-865, 2020 Dec.
Article in English | MEDLINE | ID: mdl-33197035

ABSTRACT

Spontaneous in vitro hatching of human blastocysts starts with the formation of a tunnel through the zona pellucida (ZP) by cellular projections of trophoblast cells. Our aim was to identify the proteins that are upregulated in these initially hatching cells as compared to trophectoderm (TE) cells from blastocysts that had not yet hatched. Forty seven women that underwent assisted reproduction treatment donated their ICSI-derived polyploid blastocysts for the study. In polyploid blastocysts that started spontaneous hatching, hatched clusters of cells were collected from the outer side of the ZP. Liquid chromatography mass spectrometry was applied to determine the proteins that were upregulated in these cells as compared to TE cells obtained from inside the ZP. Whole non-hatched polyploid blastocysts were used as controls. Overall 1245 proteins were identified in all samples. Forty nine proteins were significantly upregulated in hatching cells and 17 in the TE cells. There was minimal overlap between hatching and TE samples; only serine protease inhibitors (SERPINS) and lipocalin were detected in both samples. Myosin and actin were highly upregulated in the hatching cells as well as paraoxonase, N-acetylmuramoyl alanine amidase, and SERPINS clade A and galectin. In the TE cells, gamma butyrobetaine dioxygenase, lupus La protein, sialidase, lysosomal Pro-X carboxypeptidase, phospholipase b, and SERPINS clade B and A were among the most highly upregulated proteins. These findings may contribute to the basic knowledge of the molecular behavior of the specific cells that actively perforate the glycoprotein matrix of the ZP.


Subject(s)
Blastocyst/cytology , Proteomics/methods , Cell Aggregation , Ectoderm/cytology , Humans , Mass Spectrometry , Up-Regulation
2.
Isr Med Assoc J ; 20(10): 604-607, 2018 Oct.
Article in English | MEDLINE | ID: mdl-30324775

ABSTRACT

BACKGROUND: Sepsis is a common cause of hospitalization, particularly in intensive care units (ICUs), and is a major cause of morbidity and mortality. Diagnosis is often difficult due to the absence of characteristic clinical signs (e.g., fever and leukocytosis); therefore, additional markers, in addition to C-reactive protein (CRP) and white blood cell (WBC) count, are needed. OBJECTIVES: To prospectively link resting energy expenditure (REE) with CRP, WBC count, and sequential organ failure assessment (SOFA) scores in ICU patients. Such a correlation may suggest REE measurement as an additional parameter for sepsis diagnosis. METHODS: Our study comprised 41 ventilated consecutive patients > 18 years of age. Patient demographic data, height, actual body weight, and SOFA scores were collected at admission. REE was measured by indirect calorimetry. REE, CRP, and WBC measurements were collected at admission, on day three after admission, and 1 week later or as clinically indicated. RESULTS: Comparison of the REE and CRP changes revealed a significant correlation between REE and CRP changes (r = 0.422, P = 0.007). In addition, CRP changes also correlated with the changes in REE (r = 0.36, P = 0.02). Although no significant correlations in REE, WBC count, and SOFA score were found, a significant trend was observed. CONCLUSIONS: To the best of our knowledge, this is the first study to link REE and CRP levels, indicative of severe infection. Further study is needed to establish these findings.


Subject(s)
C-Reactive Protein/metabolism , Energy Metabolism/physiology , Intensive Care Units , Respiration, Artificial , Sepsis/epidemiology , Acute Disease , Aged , Aged, 80 and over , Biomarkers/metabolism , Calorimetry, Indirect/methods , Female , Humans , Leukocyte Count , Male , Middle Aged , Organ Dysfunction Scores , Prospective Studies
3.
Fertil Steril ; 106(6): 1386-1391, 2016 Nov.
Article in English | MEDLINE | ID: mdl-27567431

ABSTRACT

OBJECTIVE: To evaluate the ability to predict pregnancy outcomes of single-blastocyst transfers by measuring the ratio of inner cell mass (ICM) diameter to blastocyst diameter using time-lapse images. DESIGN: Retrospective cohort study. SETTING: University-affiliated medical center. PATIENT(S): One hundred twenty-seven women undergoing a total of 129 blastocyst transfers with intracytoplasmic sperm injection. INTERVENTION(S): Embryo monitoring by time-lapse microscopy. MAIN OUTCOME MEASURE(S): The ratio of ICM diameter to blastocyst diameter in single-blastocyst transfers and clinical pregnancy rates. RESULT(S): In phase I of the study, 63 women underwent 65 single blastocyst transfers that resulted in 25 pregnancies (40% of the women). The successfully implanted blastocysts had an average ICM/blastocyst diameter ratio of 0.487 ± 0.086, whereas the average ICM/blastocyst ratio of nonimplanted blastocysts was significantly lower (0.337 ± 0.086). The live-birth rate was 29% (18/63). In phase II, 64 single-blastocyst transfers were performed in 64 women. The ICM/blastocyst diameter ratio was measured, and blastocysts with the highest ratios were chosen for transfer. Forty-three women (67%) with an average ICM/blastocyst diameter ratio of 0.46 achieved pregnancy, and 36 of the 43 pregnancies (84%) resulted in the delivery of a healthy baby. In the 21 women (33%) who failed to achieve pregnancy, the average ICM/blastocyst ratio was 0.45. The resultant positive predictive value was 74%, and the negative predictive value was 70%. CONCLUSION(S): The ICM-to-blastocyst diameter ratio is a predictor of implantation and live birth in single-blastocyst transfers, offering a simple, noninterfering method to select blastocysts with high developmental capacity.


Subject(s)
Blastocyst Inner Cell Mass/cytology , Blastocyst/cytology , Infertility/therapy , Single Embryo Transfer , Sperm Injections, Intracytoplasmic , Time-Lapse Imaging , Academic Medical Centers , Adult , Embryo Culture Techniques , Embryo Implantation , Female , Fertility , Humans , Infertility/diagnosis , Infertility/physiopathology , Live Birth , Predictive Value of Tests , Pregnancy , Pregnancy Rate , Retrospective Studies , Single Embryo Transfer/adverse effects , Sperm Injections, Intracytoplasmic/adverse effects , Treatment Outcome
4.
J Assist Reprod Genet ; 32(12): 1811-5, 2015 Dec.
Article in English | MEDLINE | ID: mdl-26537929

ABSTRACT

PURPOSE: This is a retrospective analysis of the morphokinetics, prevalence, and implantation potential of embryos with irregular first and second cleavages as identified by time-lapse microscopy. METHODS: The study included 253 women who underwent 387 assisted reproduction treatments with intracytoplasmic sperm injection (ICSI). Each patient was assigned to one of three groups based on embryo cleavage results. In group I, one to two embryos per cycle showed irregular cleavage; group II, at least three embryos with abnormal cleavage; and in group III (the control group), all embryos cleaved normally. The number of embryos that cleaved from 1 to ≥3 cells or from 2 to ≥5 cells for each patient was recorded. Their prevalence and association with women's characteristics and pregnancy outcome were evaluated. RESULTS: The prevalence of irregular cleavage was 15.6 % among 1772 ICSI embryos. In 101 cycles, 1-2 embryos per cycle showed irregular cleavage (group I). In 32 cycles, at least 3 embryos showed abnormal cleavage (group II). In 254 cycles, all embryos cleaved normally (group III). The average age of the women in group II was significantly lower in comparison with groups I and III (32.5 ± 4.2 vs. 35.1 ± 4.9 and 35.5 ± 5.1, respectively, p < 0.02). In comparison of groups I and II, the odds ratio for ≥3 embryos with irregular cleavage in women younger than 35 was 3.48 (95 % CI, 1.28 to 9.46). Embryos with irregular cleavage were transferred in 16 women. Three live births were achieved following the transfer of single blastocysts derived from embryos with irregular cleavage from two to five cells. CONCLUSIONS: Early embryos with irregular cleavage are significantly more prevalent in younger women. When these embryos develop to the blastocyst stage, they may have normal implantation potential, leading to the birth of healthy babies.


Subject(s)
Blastocyst/cytology , Cleavage Stage, Ovum/cytology , Adult , Blastocyst/ultrastructure , Cell Division , Cleavage Stage, Ovum/ultrastructure , Embryo Culture Techniques , Embryo Implantation , Embryo Transfer , Female , Humans , Pregnancy , Pregnancy Outcome , Pregnancy Rate , Retrospective Studies
7.
Hum Reprod ; 19(1): 104-6, 2004 Jan.
Article in English | MEDLINE | ID: mdl-14688165

ABSTRACT

BACKGROUND: The complex regulation of endometrial receptivity and embryo implantation involves cytokines, several of which are stimulators of the acute-phase reactant C-reactive protein (CRP). The purpose of this study was to evaluate the concentrations of serum CRP in women treated by IVF. METHODS: Seventy-two women who underwent IVF treatment were prospectively studied. The levels of serum CRP were evaluated on the following days: oocyte retrieval, embryo transfer, 5, 6 or 7 days afterwards and 12 days after embryo transfer. RESULTS: CRP levels increased from 6.8 +/- 9.5 mg/l on oocyte retrieval day to 14.6 +/- 12.5 mg/l on days 5-7 post-transfer (P<0.0001). The ratios of CRP levels for transfer day/pick-up day were 1.2 in women who conceived versus 2.5 in the non-pregnant group (P=0.01). CONCLUSION: In women treated by IVF the concentrations of CRP in blood increase significantly during the first week following oocyte retrieval. Successful outcome is associated with a relative small increment in CRP on the day of embryo transfer.


Subject(s)
C-Reactive Protein/metabolism , Fertilization in Vitro , Adult , Embryo Transfer , Female , Fertilization , Humans , Oocytes , Osmolar Concentration , Pregnancy , Prospective Studies , Time Factors , Tissue and Organ Harvesting
8.
Clin Biochem ; 36(3): 229-31, 2003 May.
Article in English | MEDLINE | ID: mdl-12726934

ABSTRACT

OBJECTIVES: Positive readings of blood in dipstick urinalysis may indicate trauma or imbalance in hemostasis associated with drug treatment. We evaluated the possibility that the presence of semen in urine may cause false-positive hematuria. DESIGN AND METHODS: Semen specimens obtained from 25 healthy men were directly applied on urine dipsticks for evaluation of the presence of blood. Isolated sperm cells and seminal fluid were also tested. Dipstick analyses were further performed with semen samples diluted in normal urine. Four healthy male volunteers provided urine samples before and immediately after having had sexual relationships. These samples were dipstick tested for the presence of blood. RESULTS: Semen, spermatozoa and seminal fluid gave false positive results for microhematuria following direct application of samples on dipsticks as well as after their suspension in urine (p < 0.00001).Three out of four postcoital urine specimens yielded positive results for blood. CONCLUSIONS: In men, postcoital urine may be falsely "positive" for microhematuria. This may have implication on the management of male patients in emergency situations such as acute coronary syndromes.


Subject(s)
Hematuria/diagnosis , Reagent Strips , Semen , Spermatozoa , Adult , Coitus , False Positive Reactions , Hematuria/urine , Humans , Male , Middle Aged , Reproducibility of Results , Urinalysis
9.
Am Heart J ; 145(2): 248-53, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12595841

ABSTRACT

BACKGROUND: Elevation of C-reactive protein (CRP), among other markers of inflammation, is associated with an increased risk for cardiac events in patients with known coronary diseases and in apparently healthy individuals. Moreover, in patients with acute coronary syndromes, elevated serum levels of CRP are strongly predictive of the risk for death from cardiac causes. The purpose of this study was to investigate whether mechanical rupture of an atherosclerotic coronary plaque during elective stent implantation in patients with stable coronary artery disease (CAD) at low risk will cause a significant increase in serum levels of CRP. METHODS AND RESULTS: We measured serum CRP levels in 40 patients. Group 1 consisted of 12 consecutive patients with stable coronary disease who were at low risk, before and after elective coronary stent implantation. We compared the results in these patients to those of patients in 2 control groups: group 2 consisted of 12 consecutive patients with non-ST-segment elevation acute coronary syndrome (NSTSE ACS) who were undergoing coronary stent implantation, and group 3 included 16 consecutive patients with stable or unstable CAD who were undergoing diagnostic coronary angiography only without PCI. Peripheral blood samples for CRP level testing were withdrawn before percutaneous coronary intervention or angiography at the completion of the procedure, and 6, 20, and 48 hours thereafter. All patients with stable CAD (group 1) had a significant and uniform increase in serum CRP levels after elective stent implantation. The low mean baseline serum CRP levels increased 4.9 +/- 4.1-fold 20 hours after coronary intervention (2.1 +/- 1.2 before, 7.9 +/- 3.4 after, P <.002). The baseline CRP level was much higher in the patients with unstable coronary syndromes (group 2). In this group, only a 2.1-fold increase in mean CRP level was observed after stent implantation (7.4 +/- 5.5 before, 14.1 +/- 9.6 after, P <.004). Also, the response in this group was less uniform when compared with that in the stable CAD group. By contrast, in patients undergoing diagnostic coronary angiography, the mean baseline CRP level was higher than in the patients in the group with stable CAD; however, the mean CRP after the procedure was not significantly elevated in this group (4.5 +/- 3.6 before, 5.5 +/- 3.7 20 hours after, P = not significant). CONCLUSIONS: Mechanical disruption of an atherosclerotic coronary plaque during elective coronary stent implantation in patients with stable CAD who are at low risk causes a systemic inflammatory response expressed by marked elevation in CRP concentration.


Subject(s)
C-Reactive Protein/metabolism , Coronary Disease/blood , Coronary Disease/therapy , Stents , Aged , Angina Pectoris/blood , Angina Pectoris/therapy , Angioplasty, Balloon, Coronary , Anticoagulants/therapeutic use , Aspirin/therapeutic use , Biomarkers/blood , Coronary Angiography , Female , Heparin/therapeutic use , Humans , Male , Middle Aged , Statistics, Nonparametric , Time Factors
10.
Eur J Obstet Gynecol Reprod Biol ; 106(2): 148-50, 2003 Feb 10.
Article in English | MEDLINE | ID: mdl-12551781

ABSTRACT

OBJECTIVES: To evaluate coagulation parameters in healthy pregnant grand grand multiparous women. STUDY DESIGN: Prothrombin time (PT), activated partial thromboplastin time (PTT), fibrinogen, D-dimer, albumin and blood lipids were prospectively determined in 50 pregnant grand grand multiparas (> or =9 deliveries) and 50 age-matched pregnant women with low parity (< or =3 deliveries). RESULTS: The average concentrations of fibrinogen in the grand grand multiparas were 4.0+/-0.6 as compared to 4.7+/-0.7 g/l in the control group (P<0.001). In both groups D-dimer levels were >1000 ng/ml and the concentrations of albumin, triglycerides, cholesterol, high-density lipoprotein (HDL) were not significantly different. CONCLUSIONS: In healthy pregnant grand grand multiparous women fibrinogen levels are lower than in age-matched women with low parity. These changes are not directly associated with lipids metabolism during pregnancy.


Subject(s)
Fibrinogen/metabolism , Parity/physiology , Adult , Cholesterol/blood , Female , Fibrin Fibrinogen Degradation Products/metabolism , Hemoglobins/metabolism , Humans , Leukocyte Count , Partial Thromboplastin Time , Pregnancy , Prospective Studies , Prothrombin Time , Serum Albumin/metabolism
11.
Hum Reprod ; 17(12): 3170-2, 2002 Dec.
Article in English | MEDLINE | ID: mdl-12456619

ABSTRACT

BACKGROUND: Electroejaculation has become an accepted form of semen procurement in men suffering from anejaculation. However, sperm in these ejaculates often exhibit low motility. The aetiology of asthenozoospermia remains uncertain. It may be related to primary low-grade motility of sperm from anejaculatory men or to detrimental effects of the electroejaculation process itself. The aim of this study was to determine whether the procedure of electroejaculation alters the quality of the semen by comparing the characteristics of the electroejaculates with those of the normal ejaculates of the same patients. METHODS: Nine men suffering from ejaculation disorders underwent 14 procedures of electroejaculation. The sperm concentration and motility of the 14 samples from electroejaculation were compared with 20 collected at a later date after successful ejaculation by the same patients. RESULTS: The mean +/- SD concentration of the electroejaculates was (52.3 +/- 42.1)x10(6)/ml and of the natural samples (67.4 +/- 38.1)x10(6)/ml. No statistical difference was found between the two groups. The mean percentage of sperm showing progressive motility following electroejaculation was 23.1 +/- 18.8% and 36.3 +/- 16.7% after spontaneous ejaculation. No statistical difference was found between the groups (P = 0.082). Likewise, the quality of sperm motility was similar after both methods of procurement. CONCLUSIONS: In our small-sized study group (nine men with ejaculation disorders), the concentration and the motility of sperm obtained by electroejaculation were not significantly different from sperm obtained naturally.


Subject(s)
Ejaculation , Electric Stimulation , Erectile Dysfunction/physiopathology , Semen/physiology , Adult , Erectile Dysfunction/therapy , Humans , Male , Sperm Count , Sperm Motility
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