Genetic and Epidemiological Similarities, and Differences Between Postoperative Intraperitoneal Adhesion Development and Other Benign Fibro-proliferative Disorders.

Intraperitoneal adhesions complicate over half of abdominal-pelvic surgeries with immediate, short, and long-term sequelae of major healthcare concern. The pathogenesis of adhesion development is similar to the pathogenesis of wound healing in all tissues, which if unchecked result in production of fibrotic conditions. Given the similarities, we explore the published literature to highlight the similarities in the pathogenesis of intra-abdominal adhesion development (IPAD) and other fibrotic diseases such as keloids, endometriosis, uterine fibroids, bronchopulmonary dysplasia, and pulmonary, intraperitoneal, and retroperitoneal fibrosis. Following a literature search using PubMed database for all relevant English language articles up to November 2020, we reviewed relevant articles addressing the genetic and epidemiological similarities and differences in the pathogenesis and pathobiology of fibrotic diseases. We found genetic and epidemiological similarities and differences between the pathobiology of postoperative IPAD and other diseases that involve altered fibroblast-derived cells. We also found several genes and single nucleotide polymorphisms that are up- or downregulated and whose products directly or indirectly increase the propensity for postoperative adhesion development and other fibrotic diseases. An understanding of the similarities in pathophysiology of adhesion development and other fibrotic diseases contributes to a greater understanding of IPAD and these disease processes. At a very fundamental level, blocking changes in the expression or function of genes necessary for the transformation of normal to altered fibroblasts may curtail adhesion formation and other fibrotic disease since this is a prerequisite for their development. Similarly, applying measures to induce apoptosis of altered fibroblast may do the same; however, apoptosis should be at a desired level to simultaneously ameliorate development of fibrotic diseases while allowing for normal healing. Scientists may use such information to develop pharmacologic interventions for those most at risk for developing these fibrotic conditions.

Is There a Genetic Predisposition to Postoperative Adhesion Development?

Adhesions are permanent fibrovascular bands between peritoneal surfaces, which develop following virtually all body cavity surgeries. The susceptibility to develop, and the severity, of adhesions following intra-abdominal surgery varies within and between individuals, suggesting that heritable factors influence adhesion development. In this manuscript, we discuss the pathophysiology of adhesion development from the perspective of genetic susceptibility. We restrict our discussion to genes and single-nucleotide polymorphisms (SNPs) that are specifically involved in, or that cause modification of, the adhesion development process. We performed a literature search using the PubMed database for all relevant English language articles up to March 2020 (n = 186). We identified and carefully reviewed all relevant articles addressing genetic mutations or single-nucleotide polymorphisms (SNPs) that impact the risk for adhesion development. We also reviewed references from these articles for additional information. We found several reported SNPs, genetic mutations, and upregulation of messenger RNAs that directly or indirectly increase the propensity for postoperative adhesion development, namely in genes for transforming growth factor beta, vascular endothelial growth factor, interferon-gamma, matrix metalloproteinase, plasminogen activator inhibitor-1, and the interleukins. An understanding of genetic variants could provide insight into the pathophysiology of adhesion development. The information presented in this review contributes to a greater understanding of adhesion development at the genetic level and may allow modification of these genetic risks, which may subsequently guide management in preventing and treating this challenging complication of abdominal surgery. In particular, the information could help identify patients at greater risk for adhesion development, which would make them candidates for anti-adhesion prophylaxis. Currently, agents to reduce postoperative adhesion development exist, and in the future, development of agents, which specifically target individual genetic profile, would be more specific in preventing intraperitoneal adhesion development.

Glyphosate Induces Metaphase II Oocyte Deterioration and Embryo Damage by Zinc Depletion and Overproduction of Reactive Oxygen Species.

Glyphosate is the most popular herbicide used in modern agriculture, and its use has been increasing substantially since its introduction. Accordingly, glyphosate exposure from food and water, the environment, and accidental and occupational venues has also increased. Recent studies have demonstrated a relationship between glyphosate exposure and a number of disorders such as cancer, immune and metabolic disorders, endocrine disruption, imbalance of intestinal flora, cardiovascular disease, and infertility; these results have given glyphosate a considerable amount of media and scientific attention. Notably, glyphosate is a powerful metal chelator, which could help explain some of its effects. Recently, our findings on 2,3-dimercapto-1-propanesulfonic acid, another metal chelator, showed deterioration of oocyte quality. Here, to generalize, we investigated the effects of glyphosate (0 - 300 µM) on metaphase II mouse oocyte quality and embryo damage to obtain insight on its mechanisms of cellular action and the tolerance of oocytes and embryos towards this chemical. Our work shows for the first time that glyphosate exposure impairs metaphase II mouse oocyte quality via two mechanisms: 1) disruption of the microtubule organizing center and chromosomes such as anomalous pericentrin formation, spindle fiber destruction and disappearance, and defective chromosomal alignment and 2) substantial depletion of intracellular zinc bioavailability and enhancement of reactive oxygen species accumulation. Similar effects were found in embryos. These results may help clarify the effects of glyphosate exposure on female fertility and provide counseling and preventative steps for excessive glyphosate intake and resulting oxidative stress and reduced zinc bioavailability.

Successful Oocyte Retrieval, Fertilization, and Clinical Pregnancy with Low Serum ß-hCG on the Day of Oocyte Collection: A Reappraisal of the Definition of the Empty Follicle Syndrome.

OBJECTIVE: To describe a case of successful oocyte retrieval, fertilization and clinical pregnancy despite very low ß-hCG level, twelve hours after ovulation trigger. DESIGN: Case report. Setting. Academic medical center. Patient. A 38-year-old patient inadvertently administered 2,000 IU hCG for final oocyte maturation; serum hCG twelve hours later was 16 IU/L. Interventions. Effort to obtain and administer a booster dose of hCG over the next twenty-seven hours failed. Main Outcome. Successful oocyte retrieval. RESULTS: Fourteen oocytes were retrieved of which twelve were in metaphase II and nine fertilized after intracytoplasmic sperm injection (ICSI). Of these, eight embryos survived to day 5 and were subjected to preimplantation genetic screening (PGS) by comparative genomic hybridization (CGH). Results were available the next day, three of the embryos were euploid and one was transferred on day 6. Pregnancy was confirmed twelve days later and currently the patient has an ongoing singleton intrauterine pregnancy. CONCLUSION: Reproductive Endocrinology and Infertility specialists should be aware that final oocyte maturation could occur following injection of a lower dose of hCG with excellent fertilization rate and embryo development.

Hypochlorous acid reversibly inhibits caspase-3: a potential regulator of apoptosis.

Caspase-3 is involved in apoptosis. Here, we examine whether hypochlorous acid (HOCl), a final product of myeloperoxidase (MPO), is a modulator of caspase-3 at relatively low concentrations and also its application on metaphase II mouse oocytes. We utilised caspase-3 activity assay, TUNEL assay, the CellEvent caspase 3/7 fluorescent assay, and the MPO/hydrogen peroxide (H2O2) system on mouse oocytes with and without cumulus cells to examine whether low concentrations of HOCl mediate apoptosis by inhibition of caspase-3. A UV-visible spectrophotometer was used to study caspase-3 activity. To determine whether HOCl mediates apoptosis in mouse oocytes, two different concentrations (10 and 100 µM) of HOCl generated by the MPO/H2O2 system were used as treatments (10 µM had little effect on oocyte quality, while 100 µM showed significant deterioration). Induction of apoptotic cell death was determined by TUNEL Assay and the CellEvent caspase 3/7. HOCl mediates caspase-3 inactivation in a dose dependent manner. Subsequent addition of dithiothreitol caused recovery of caspase-3 activity indicating involvement of the oxidation of the Cys-thiol group. Accumulation of HOCl generated by MPO in the presence of caspase-3 also inhibits MPO but requires higher HOCl concentrations, indicating specificity of lower HOCl concentrations to inhibition of caspase-3. Exposure of oocytes to lower HOCl concentrations generated by MPO-H2O2 system prevents MPO-mediated apoptosis whereas exposure to higher HOCl (100 µM) showed apoptosis. Similar results were observed by using the CellEvent caspase 3/7 assay. Low concentrations of HOCl inhibit caspase-3 activity, and may play a role in regulating apoptosis, thus affecting oocyte quality.HighlightsCaspase-3 is involved in apoptosis pathway and loss of this regulation is seen in several diseases.These conditions are associated with inflammation and higher myeloperoxidase (MPO) activity.We examined whether hypochlorous acid (HOCl), generated by MPO, is a modulator of caspase-3.Caspase-3 activity showed a dose dependent decrease with HOCl and this reaction was reversible.HOCl modulates caspase-3 activity and may play a physiological role in regulating apoptosis.

Catalase prevents myeloperoxidase self-destruction in response to oxidative stress.

Catalase (CAT) and myeloperoxiase (MPO) are heme-containing enzymes that have attracted attention for their role in the etiology of numerous respiratory disorders such as cystic fibrosis, bronchial asthma, and acute hypoxemic respiratory failure. However, information regarding the interrelationship and competition between the two enzymes, free iron accumulation, and decreased levels of non-enzymatic antioxidants at sites of inflammation is still lacking. Myeloperoxidase catalyzes the generation of hypochlorous acid (HOCl) from the reaction of hydrogen peroxide (H2O2) and chloride (Cl-). Self-generated HOCl has recently been proposed to auto-inhibit MPO through a mechanism that involves MPO heme destruction. Here, we investigate the interplay of MPO, HOCl, and CAT during catalysis, and explore the crucial role of MPO inhibitors and HOCl scavengers in protecting the catalytic site from protein modification of both enzymes against oxidative damage mediated by HOCl. We showed that CAT not only competes with MPO for H2O2 but also scavenges HOCl. The protective role provided by CAT versus the damaging effect provided by HOCl depends in part on the ratio between MPO/CAT and the affinity of the enzymes towards H2O2 versus HOCl. The severity of such damaging effects mainly depends on the ratio of HOCl to enzyme heme content. In addition to its effect in mediating protein modification and aggregation, HOCl oxidatively destroys the catalytic sites of the enzymes, which contain porphyrin rings and iron. Thus, modulation of MPO/CAT activities may be a fundamental feature of catalysis, and functions to down-regulate HOCl synthesis and prevent hemoprotein heme destruction and/or protein modification.

Acrolein, a commonly found environmental toxin, causes oocyte mitochondrial dysfunction and negatively affects embryo development.

Recent studies have revealed that acrolein, a commonly found toxin and a potent metabolite of cyclophosphamide (CTX), can cause deterioration of mouse oocyte quality through a mechanism involving the generation of reactive oxygen species (ROS). We extend these studies to evaluate the effects of acrolein, in varying concentrations, on the oocyte mitochondrial membrane and oocyte apoptosis and its effect on embryo development in vitro. Metaphase II mouse oocytes were exposed for 45 minutes to acrolein and CTX (10 & 25 µM) and mitochondrial dysfunction, a major source of ROS overproduction, was evaluated by the 5,5,6,6-tetrachloro-1,1,3,3-tetraethyl-ß-benzimidazolylcarbocyanine iodide (JC-10) mitochondrial membrane potential assay. Treatment with acrolein led to mitochondrial membrane damage as well as induction of apoptosis compared to untreated control (p < 0.05). Similar results were obtained when oocytes were exposed to CTX (p < .05). Subsequently, the effect of acrolein exposure was evaluated by observing in vitro development of embryos after exposure. Acrolein treatment caused higher proportions of arrested and poor-quality embryos, evidenced by irregular cleavage, severe asymmetry of blastomeres, presence of large percentage of anuclear fragments, and dark granularity of the cytoplasm. Development at various durations in culture revealed that optimal embryo growth was significantly inhibited in a dose dependent manner, when compared to control (p < .05). A global model that links acrolein accumulation, generation of ROS, and mitochondrial dysfunction and their effect on oocyte and embryo quality is discussed further. Collectively, understanding the mechanism by which CTX and acrolein impact fertility is helpful in finding potential alternative or supplemental treatment options.

Melatonin prevents hypochlorous acid-mediated cyanocobalamin destruction and cyanogen chloride generation.

Hypochlorous acid (HOCl) is a potent cytotoxic oxidant generated by the enzyme myeloperoxidase (MPO) in the presence of hydrogen peroxide (H2 O2 ) and chloride (Cl- ). Elevated levels of HOCl play an important role in various pathological conditions through oxidative modification of several biomolecules. Recently, we have highlighted the ability of HOCl to mediate the destruction of the metal-ion derivatives of tetrapyrrole macrocyclic rings such as hemoproteins and vitamin B12 (VB12 ) derivatives. Destruction of cyanocobalamin, a common pharmacological form of VB12 mediated by HOCl, results in the generation of toxic molecular products such as chlorinated derivatives, corrin ring cleavage products, the toxic blood agents cyanide (CN- ) and cyanogen chloride (CNCl), and redox-active free cobalt. Here, we show that melatonin prevents HOCl-mediated cyanocobalamin destruction, using a combination of UV-Vis spectrophotometry, high-performance liquid chromatography analysis, and colorimetric CNCl assay. Identification of several melatonin oxidation products suggests that the protective role of melatonin against HOCl-mediated cyanocobalamin destruction and subsequent CNCl generation is at the expense of melatonin oxidation. Collectively, this work highlights that, in addition to acting as an antioxidant and as a MPO inhibitor, melatonin can also prevent VB12 deficiency in inflammatory conditions such as cardiovascular and neurodegenerative diseases, among many others.

Association of the very early rise of human chorionic gonadotropin with adverse outcomes in singleton pregnancies after in vitro fertilization.

OBJECTIVE: To determine if very early serum hCG, a marker of trophoblast differentiation, is associated with adverse perinatal outcomes in singleton pregnancies. DESIGN: Retrospective cohort study. SETTING: University fertility program. PATIENT(S): A total of 360 singleton IVF live births. INTERVENTION(S): Serial hCG measurements were used to determine the within-woman slope for hCG (hCG rise). MAIN OUTCOMES MEASURE(S): Primary outcomes included birth weight and gestational age at delivery. Statistical comparisons used t test, chi-square test, and linear and logistic regressions as appropriate. RESULT(S): hCG rise was positively associated with birth weight but not gestational age at delivery. Infant sex, gestational age, and type of embryo transfer (fresh vs. frozen/thawed) were significantly associated with birth weight and confounded the associations of interest. hCG rise was slower among subjects delivering an infant with low birth weight (slope 0.386 ± 0.05 vs. 0.407 ± 0.06) or small for gestational age (slope 0.371 ± 0.07 vs. 0.406 ± 0.06). Analysis of hCG rise by quartile showed that, compared with the first quartile (slowest), subjects with a rate of hCG rise in the fourth quartile (fastest) had a significantly decreased risk of delivering an infant of low birth weight. No relationship was noted between hCG rise and hypertensive disorders of pregnancy. CONCLUSION(S): Slower very early first-trimester hCG rise is associated with low birth weight but not gestational age at delivery among singleton IVF conceptions. The rate of increase in serum hCG may reflect early trophoblast differentiation and placentation and, possibly, may predict subsequent development.

Isolated Recurrent Hematotrachelos After Abdominal Myomectomy A Case Report.

BACKGROUND: Isolated hematotrachelos is an ex- tremely rare condition. More commonly it is associated with hematometra and/or hematocolpos. It may devel- op secondary to congenital anomalies of the reproductive tract or may be an acquired condition following cervical surgery or manipulation. No case has been reported fol- lowing abdominal myomec- tomy. CASE: A 40-year-old, nul- ligravid woman was diag- nosed with isolated hemato- trachelos when she presented with severe abdominal pain following ovulation induction with clomiphene citrate. After the diagnosis was obtained, she recounted 2 similar presentations occurring months earlier. Both episodes had been attributed to gastroenteritis, but each had re- solved with "explosive" onset of menses. All 3 episodes plus 1 later recurrence happened within a 36-month period following an abdominal myomectomy. CONCLUSION: Acquired hematotrachelos is a rare condition, and the diagnosis is often missed. To our knowledge, this is the first reported case of isolated, recurrent hematotrachelos following the use of a uterine manipulator cannula in association with an abdominal myomectomy.

Reproductive hormones and postpartum mood disturbances in Greek women.

AIM: Postpartum mood disturbances are common among Greek women, with postpartum depression (PPD) being as high as 19%. This study aimed to investigate whether sex steroid hormone levels affect the incidence of postpartum mood disturbances. MATERIALS AND METHODS: Fifty-seven women were evaluated for postpartum mood disturbances using the Postpartum Blues Questionnaire and the Edinburgh Postnatal Depression Scale on the 1st and 6th week. Serum estradiol, progesterone and testosterone concentrations were measured upon admission for delivery and daily until the fourth postpartum day. We then studied the association between hormone levels and the scores in the two psychometric scales. RESULTS: Testosterone was the only hormone that was marginally associated with psychometric scoring in simple regression analysis. (Postpartum Blues during days 1-4: b = 4.291, 95% C.I. -0.796 to 9.377 and p-value = 0.096). Women with lower testosterone drops had higher scores in Postpartum Blues Questionnaire. This association, however, lost statistical significance in the multivariable analysis after adjusting for pregnancy duration. In multiple regression analysis, only pregnancy duration had the most constant adverse effect on psychometric scores: The shorter the duration of pregnancy, the higher the scores for Postpartum Blues. (r = -0.39, p < 0.01). CONCLUSIONS: Our findings do not indicate an association between the occurrence of postpartum mood disorders and sex steroid hormone levels. Preterm labour may be associated with a higher risk of postpartum mood disturbances.