Radical scavenging activity of Caesalpinia spinosa.

OBJECTIVES: To assess protective effects of the tara (Caesalpinia spinosa) extract against hyaluronan (HA) degradation evoked by cupric ions and ascorbate. METHODS: Uninhibited/inhibited HA degradation was assayed by a decrease in dynamic viscosity of the HA solutions, whereas as a method rotational viscometry was used. To determine radical scavenging capacity of the tara extract, the ABTS and DPPH assays were performed. RESULTS: The results of rotational viscometry showed remarkable protective effects of the tara extract against the degradation of HA. In the ABTS and DPPH assays the IC50 values of the tara extract 1.59 and 30.8 µg/mL indicated quite high radical scavenging properties. CONCLUSION: The tara extract is an efficient antioxidant as demonstrated by the methods used.

Germination Study of Some Protein-Based Gels Obtained from By-Products from the Leather Industry on Tomato and Pepper Seeds.

This study aimed to evaluate the biostimulant effects of three protein-based gels, GHC 1-B (20% gelatin (GPU-B) obtained by thermal hydrolysis from residual untanned leather and 80% collagen hydrolysates (HCE-B) obtained by alkaline-enzymatic hydrolysis from residual bovine-tanned leather), GHC 2-B (40% keratin hydrolysate (HKU-B) obtained by alkaline-enzymatic hydrolysis from sheep wool + 40% HCE-B + 20% GPU-B), and GHC 3-B (20% GPU-B + 80% hydrolyzed collagen (HPU-B) obtained by thermal and enzymatic hydrolysis from residual untanned leather). A germination study was carried out on pepper and tomato seeds at concentrations of 1%, 3%, and 10%. As a result of the study, it was found that all three protein-based gels showed a stimulatory effect on the tomato seeds at a 1% concentration, where the Gi (germination index) was ˂100%. The GHC 2-B variant had the highest stimulatory effect (Gi-190.23%). Pepper seeds have proven to be more sensitive to the gel's composition. The concentration at which it proved to be non-inhibitory (Gi-88.29%) was 1% in the case of GHC 2-B. It was found that the presence of hydrolyzed keratin in the composition can be a plus compared to the other two protein gels tested due to its composition, which is richer in phytonutrient compounds (e.g., sulfur molecules).

Innovative Protein Gel Treatments to Improve the Quality of Tomato Fruit.

This study aims to establish the effect of biostimulatory protein gels on the quality of tomato. One of the most consumed vegetables, tomato (Lycopersicon esculentum Mill.) is a rich source of healthy constituents. Two variants of protein gels based on bovine gelatin and keratin hydrolysates obtained from leather industry byproducts were used for periodical application on the tomato plant roots in the early stage of vegetation. The gels were characterized by classical physicochemical methods and protein secondary structure was obtained by FTIR band deconvolution. After ripening, tomato was analyzed regarding its content of quality indicators (sugars and organic acids) and antioxidants (lycopene, ß-carotene, vitamin C, polyphenols). The results emphasized the positive effects of the protein gels on the quality parameters of tomato fruit. An increase of 10% of dry matter and of 30% (in average) in the total soluble sugars was noted after biostimulant application. Also, lycopene and vitamin C recorded higher values (by 1.44 and 1.29 times, respectively), while ß-carotene showed no significant changes. The biostimulant activity of protein gels was correlated with their amino acid composition. Plant biostimulants are considered an ecological alternative to conventional treatments for improving plant growth, and also contributing to reduce the intake of chemical fertilizers.

Collagen and Keratin Hydrolysates to Delay the Setting of Gypsum Plaster.

Leather and wool waste represent a high concern due to the low level of valorization and circular economy demands for upcycling of biomass resources. Both biomasses can be easily processed as protein hydrolysates and used as functional additives due to the amphiphilic and tunable properties of collagen and keratin proteins. The chemical, physical, and structural investigations of collagen and keratin hydrolysate properties showed that the chelating abilities due to carboxylic groups can be exploited for gypsum retardant additives. The molecular weights and amino acid compositions of three different hydrolysates showed only slight influences on the setting time of gypsum; all three proteins delayed the setting time of gypsum between 60 and 120 min, as compared to the commercial plaster with a 30 min setting time. Higher molecular weight and more carboxylic active groups showed slight improvements in the setting time of mortars. The improved properties of keratin hydrolysate as compared to low molecular collagen hydrolysate were attributed to foaming and conductive properties. The mechanism of mortar setting delaying through calcium ions complexation by protein hydrolysates was shown by electric conductivity evolution of plasters with and without protein additives over time, supported by foaming properties, amino acid, and functional groups' composition. Lower bending strength values for the higher concentration of proteins do not reduce the potential to use the protein hydrolysates as retardant additives in mortar fabrication.

Wool Keratin Hydrolysates for Bioactive Additives Preparation.

The aim of this paper was to select keratin hydrolysate with bioactive properties by using the enzymatic hydrolysis of wool. Different proteolytic enzymes such as Protamex, Esperase, and Valkerase were used to break keratin molecules in light of bioactive additive preparation. The enzymatic keratin hydrolysates were assessed in terms of the physico-chemical characteristics related to the content of dry substance, total nitrogen, keratin, ash, cysteic sulphur, and cysteine. The influence of enzymatic hydrolysis on molecular weight and amino acid composition was determined by gel permeation chromatography (GPC) and gas chromatography-mass spectrometry (GC-MS) analyses. Antimicrobial activity of keratin hydrolysates was analysed against Fusarium spp., a pathogenic fungus that can decrease the quality of plants. The bioactivity of enzymatic hydrolysates was tested on maize plants and allowed us to select the keratin hydrolysates processed with the Esperase and Valkerase enzymes. The ratio of organised structures of hydrolysate peptides was analysed by attenuated total reflectance-Fourier transform infrared (ATR-FTIR) deconvolution of the amide I band and may explain the difference in their bioactive behaviour. The most important modifications in the ATR spectra of maize leaves in correlation with the experimentally proven performance on maize development by plant length and chlorophyll index quantification were detailed. The potential of enzymatic hydrolysis to design additives with different bioactivity was shown in the case of plant growth stimulation.

The perioperative effect of anesthetic drugs on the immune response in total intravenous anesthesia in patients undergoing minimally invasive gynecological surgery.

BACKGROUND: The specific mechanism of action of each anesthetic drug on the immune system is still incompletely known. It is important to know how the various anesthetics used in minimally invasive surgery (MIS) act on the inflammatory response because the choice of the anesthetic agent can influence the patient's immune system. AIM: Evaluation of the effect of anesthetic drugs used for total intravenous anesthesia (Propofol and Midazolam) on the inflammatory response after minimally invasive gynecological surgery. PATIENTS, MATERIALS AND METHODS: The inflammatory response in 20 female patients who underwent minimally invasive gynecological surgery under which intravenous anesthesia was performed. Depending on the combination of anesthetics used, we subdivided the study group into two groups, Group 1 consisting of the patients (n=10) who were given for total intravenous anesthesia, the combination with Midazolam+Fentanyl, and Group 2 (n=10) the patients who received the combination of Propofol+Fentanyl, respectively. Surgical interventional procedures included day surgery: diagnostic and operative hysteroscopy, endometrial ablation, surgical treatment of vulvar disorders. Serological profiling of patients was performed by dosing the serum concentration of nucleotide-binding domain (NOD) and leucine-rich repeat protein 3 (NLRP3) inflammasomes, interleukin (IL)-6, tumor necrosis factor-alpha (TNF-α), IL-10 before and two hours after the surgical procedure. RESULTS: In our study, we found that in both groups of patients (Midazolam+Fentanyl - Group 1, Propofol+Fentanyl - Group 2), NLRP3 and cytokines concentrations in the serum were higher after MIS than those before MIS. CONCLUSIONS: It appears that both Midazolam and Fentanyl and Propofol and Fentanyl have an immunomodulatory action due to the anti-inflammatory effect of both anesthetics. Therefore, anesthesiologists must choose an anesthetic method that uses individualized anesthetic agents, depending on the patient's immune status and disease.

Reversal of brain aging by targeting telomerase: A nutraceutical approach.

Telomeres, the protective caps of chromosomes, shorten with age, as telomerase, the enzyme responsible for the compensation of telomere erosion, is inactive in the majority of cells. Telomere shortening and subsequent cell senescence lead to tissue aging and age­related diseases. Neurodegenerative disorders, characterized by the progressive loss of neurons among other hallmarks of aged tissue, and poor cognitive function, have been associated with a short telomere length. Thus, telomerase activity has emerged as a therapeutic target, with novel agents being under investigation. The present study aimed to examine the effects of a novel natural telomerase activator, 'Reverse™', containing Centella asiatica extract, vitamin C, zinc and vitamin D3 on the brains of 18­month­old rats. The administration of the 'Reverse™' supplement for 3 months restored telomerase reverse transcriptase (TERT) expression in the brains of rats, as revealed by ELISA and immunohistochemistry. In addition, the findings from PCR­ELISA demonstrated an enhanced telomerase activity in the cerebellum and cortex cells in the brains of rats treated with the 'Reverse™' supplement. The histopathological findings confirmed a structural reversibility effect close to the differentiation observed in the young control group of rats treated with two capsules/kg body weight of the 'Reverse™' supplement. On the whole, the findings of the present study provide a strong indication that an increased telomerase activity and TERT expression may be achieved not only in the postnatal or embryonic period, but also in the brains of middle­aged rats through nutraceutical supplementation. The use of the 'Reverse™' supplement may thus contribute to the potential alleviation of a number of central nervous system diseases.

The utility of p16, E-cadherin and Ki67 in cervical squamous intraepithelial lesions diagnosis.

In this study, we included 26 cases diagnosed as squamous intraepithelial lesions, which were examined histopathologically, and in terms of p16, E-cadherin and Ki67 immunoexpression. In low-grade lesions, p16 expression was limited to one third below the epithelium, E-cadherin has a membranous pattern and Ki67 proliferation index had low values. In high-grade lesions, the p16 diffuse stain was present in two thirds or all epithelium layers, E-cadherin expression became aberrant, with membranous and cytoplasmic pattern and Ki67 proliferation index was high. These biomarkers have proven useful to accurately assess the extent of lesions and to identify lesions with high risk of progression.

Ovarian ectopic pregnancy: the role of complex morphopathological assay. Review and case presentation.

Ovarian ectopic pregnancy (OEP) represents the rarest type of ectopic pregnancy, accounting for 1-3% of this pathology. The diagnosis of this pathology is challenging due to the non-specific clinical aspects and the ultrasound examination hampered by the lack of visible gestational sac in the presence of hematocele and hemoperitoneum. The purpose of the extended histopathological (HP) examination was to identify particular aspects of the OEP trophoblast and to highlight potential local ovarian modifications which can determine pregnancy fixation at this level. The patient presented local favorable conditions for intraovarian nidation, conditions confirmed by the HP classical examination and by the immunohistochemical evaluation. We identified, using classical Hematoxylin-Eosin, Masson's trichrome and Periodic Acid-Schiff (PAS)-Hematoxylin, necrotic hemorrhage, accentuated vascular thrombosis and high density lymphoplasmocytary infiltrate. These modifications increased local adhesivity and cell destruction through hypoperfusion. Anti-cluster of differentiation antibodies (CD34, CD38, tryptase) revealed the low number of intravillous vessels and the high number of macrophages and mastocytes involved in the local inflammatory process heighten. We identified the presence of trophoblast tissue in the ovarian structure using anti-cytokeratin AE1∕AE3 (CK AE1∕AE3)/anti-cytokeratin 7 (CK7) antibodies. The anti-alpha-smooth muscle actin (α-SMA) and anti-vimentin (VIM) antibodies displayed the density of myofibroblasts and intravillous stromal cells and with the aid of anti-progesterone receptor (PR) antibody, we identified the corpus luteum hormonal response in the OEP. The placental villosities present a blocked multiplication process at the anti-apoptotic B-cell lymphoma 2 (BCL2) protein, confirmed by the Ki67 cell proliferation and tumor protein 63 (p63) immunomarkers. Anti-neuron specific enolase (NSE), anti-calretinin and anti-inhibin A antibodies showed the particular aspects of the granulosa and internal theca cells, which may be involved in oocyte release blockage, intraluteal and extraluteal fecundation of the OEP.

New Nanofibers Based on Protein By-Products with Bioactive Potential for Tissue Engineering.

Concentrated collagen hydrolysate (HC10CC), rabbit collagen glue (RCG), and keratin hydrolysate (KH) were investigated in terms of their extraction from mammalian by-products and processing by electrospinning. The electrospun nanofibers were characterized by scanning electron microscopy coupled with the energy dispersive X-ray spectroscopy (SEM/EDS), attenuated total reflectance Fourier transform infrared spectroscopy (ATR-FTIR), differential scanning calorimetry (DSC), and indentation tests. The cytotoxicity of the electrospun nanofibers was conducted on L929 fibroblast cells using MTT and LDH assays and cell morphology observations. The electrospun RCG and KH nanofibers morphology showed an average size of nanofibers ranging between 44 and 410 nm, while the electrospun HC10CC nanofibers exhibited higher sizes. The ATR-FTIR spectra performed both on extracted proteins and electrospun nanofibers showed that the triple helix structure of collagen is partially preserved. The results were in agreement with the circular dichroism analysis for protein extracts. Furthermore, the viscoelastic properties of electrospun KH nanofibers were superior to those of electrospun RCG nanofibers. Based on both in vitro quantitative and qualitative analysis, the electrospun nanofibers were not cytotoxic, inducing a healthy cellular response. The results of new electrospun protein-based nanofibers may be useful for further research on bioactive properties of these nanofibers for tissue engineering.

Tuberous sclerosis complex: report of two intrafamilial cases, both in mother and daughter.

Tuberous sclerosis complex (TSC) is a multisystem syndrome characterized by neurological symptoms and tumors in multiple organs including kidney, brain, skin, eyes, heart and lung. Kidney and brain are the two most frequently affected organs in TSC. TSC is an autosomal disorder with extensive clinical variability. We described TSC in a family at a mother and her daughter. We emphasized the importance of Computed Tomography in the discovery of some asymptomatic organic involvement as bilateral renal angiolipoma in the mother.

Correlations between clinical and placental histopathological and immunohistochemical features in women with and without hereditary thrombophilia.

AIM: The primary objective of this study was to correlate hereditary thrombophilia (high- or low-risk) with specific placental histopathological (HP) and∕or immunohistochemical (IHC) changes, for confirming∕ruling out a possible linkage between these two biological parameters. PATIENTS, MATERIALS AND METHODS: We present a 3-year prospective study conducted between 2016 and 2019 that enrolled 90 women registered in two Clinics of Obstetrics and Gynecology in Craiova, Romania, with personal thrombotic and/or pathological obstetrical history. The HP and IHC analysis of the placenta was performed using monoclonal anti-cluster of differentiation 34 (CD34) antibody, anti-hypoxia-inducible factor-1 alpha (HIF-1α) and anti-endothelial nitric oxide synthase (eNOS) antibody. RESULTS: There was a high incidence of all thrombophilia (TPh) mutations in Caucasian women with thrombotic and obstetrical complications. Among them, both HP and IHC examination revealed significant changes. These were more severe in the placentas of patients with homozygous Factor V Leiden (FVL) gene mutation and double heterozygous FVL∕PII gene mutation. Multiple placental infarctions with massive fibrinoid necrosis and an increase in syncytial knots are common findings. In the same group, we found by means of IHC examination - intense positive HIF-1α and eNOS immunoexpression, and low positive CD34 expression, especially in fibrinoid necrosis and thrombosis areas. We found no correlation between clinical, HP and IHC changes in patients with low-risk TPh or without TPh. CONCLUSIONS: Among patients with obstetric and thrombotic complications, there is a high prevalence of TPh. It appears that hypercoagulability reported in high-risk thrombophilia (HR-TPh) has major effects on placental tissue (fibrinoid necrosis, multiple thromboses, hypoxia and oxidative stress). Significant placental changes were found predominantly in women with HR-TPh. Strategies for TPh screening based on HP/IHC pattern would be, most probably, more cost-effective compared with the extended TPh testing offered in large populations. This way, a smaller number of patients will be tested and in this group a higher proportion of patients will be found as having HR-TPh mutations.

Triple immunohistochemistry for assessing the inflammatory, vascular and progression of adenomyosis.

Adenomyosis is a benign pathology, common to both women at reproductive age as well as later during menopause. This condition is accompanied by a strong symptomatology, which has induced intense research on this topic. From a morphological point of view, it is represented by the existence of endometrial glands and, sometimes, of the periglandular stroma (endometriosis) in the structure of the myometrium, at a significant distance from the normal endometrium. Various inflammatory, vascular and mechanical factors accentuate the symptoms and evolution of this pathology. Our study included a total number of 32 patients, eight cases for each of the following histopathological subtypes: endometrium - proliferative phase, endometrium - secretory phase, myometrium with endometrial glands (adenomyosis), and myometrium with hyperplastic transformation of endometrial glands (hyperplastic adenomyosis), respectively. We have conducted clinical, morphological and morphopathological studies of the structures in question. Using the classical histological technique (Hematoxylin-Eosin), we identified the glandular structures; utilizing immunohistochemistry, we have labeled the endometrial epithelium with the anti-cytokeratin 7 (CK7) antibody and we analyzed the periglandular cell types of the immune system: T-lymphocytes using anti-cluster of differentiation (CD) 3 antibody, macrophages using anti-CD68 antibody, mast cells using anti-tryptase antibody, periglandular vascularization with the reaction using anti-CD34∕anti-CD31 antibodies, thus demonstrating their involvement in the development of adenomyosis. The interesting aspect of this study is the technique of simultaneously labeling of the inflammatory, vascular and epithelial elements.

Morphological and ultrasound findings in multiple pregnancy placentation.

The incidence of multiple pregnancy has significantly increased over the past decades, reaching different statistics to double, triple, or even overcome these numerical orders globally. Zygosity and chorionicity are the key elements in the multiple pregnancy but the placentation issue should be correlated primarily with zygosity, unlike chorionicity that should be correlated with the outcome and complications of multifetal gestation. Multiple pregnancy is by itself a special maternal-fetal condition, and the monochorionic one, moreover, due to specific complications. These aspects make early assessment of chorionicity and amnionicity a priority. Ultrasound is essential in pregnancy but pathological placental examination after delivery is complementary, in order to have a complete overview of potential mechanisms and pathogenesis affecting twin gestation. In this review, we highlight both ultrasound aspects specific to multifetal placentation, complemented by macro and microscopic morphological aspects, which underpin the obstetric imaging.

Status thymicolymphaticus: real or fake?

Sudden infant death syndrome (SIDS) is the sudden, unexpected death of an infant less than one year of age that remains unexplained after a full investigation. SIDS is the most frequent cause of death of infants between two weeks and one year of age, explaining 35% to 55% of all deaths in this age group. We report a newborn male who died soon after birth. The newborn was cyanotic, bradycardic at first, and then asystolic; without any vesicular murmur, apneic, low amplitude thorax movements, even under conditions of positive pressure ventilation on the endotracheal tube. The microscopic aspect thymus highlighted a corticomedullary ratio quite high in favor of the cortical, rich in lymphocyte population, with the dilated subcapsular sinuses. In this report, we considered that cardiorespiratory failure, which was the immediate cause of death, could have been caused by the thymus hypertrophy. This hypertrophy can be a complication of an intrapartum preexistent condition, most probably of hepatic nature.

Placental findings in pregnancies complicated with IUGR - histopathological and immunohistochemical analysis.

OBJECTIVE: Placental lesions and placental ischemia are typical elements of intrauterine growth restriction (IUGR). The aim of this study is to analyze histological and immunohistochemical (IHC) changes in the placentas of IUGR fetuses. MATERIALS AND METHODS: In this prospective study, 126 placentas from small for gestational age (SGA) pregnancies (newborns with birth weight <10th percentile) that formed the study group and 31 placentas from pregnancies without SGA representing control group, were included. Placentas were examined according to standard protocol. Histopathological and IHC examinations of placentas were performed for analysis. RESULTS: A certain type of lesion of placental injury is increased in placentas from SGA pregnancies. These placental lesions were placental infarction (over 5%), increased syncytial knots, intervillous fibrinoid deposition, villous thrombohematoma. Other common placental lesions were probably related to fetal adaptation to placental ischemia or represent a placental change characteristic of pregnancy evolution. CONCLUSIONS: It seems that although IUGR∕SGA fetuses are more commonly associated with histological placental abnormalities, it cannot be established whether these abnormalities certainly contribute to IUGR, as there are no specific placental lesions in SGA placentas. Pseudo-angiomatous aspect, associated with increased syncytial knots, was specific for vascular hypoxia. Especially the magnitude of modifications of the placental structure beyond the qualitative modifications, which also lead to functional changes, are involved in this pathology of pregnancy, the onset of lesions being triggered at the level of stem villi.

Clinical, morphological and immunohistochemical survey in different types of endometriosis.

Endometriosis is a benign pathology, commonly found in women at reproductive age. It is represented by the ectopic presence of the endometrial glandular epithelium in several tissues and organs. This ectopically located tissue can display premalignant or even malignant changes under the influence of certain factors that affect cell structure, function and proliferation. Our study includes a total of 28 patients, with endometriosis of different localizations: ovarian or pelvic endometriosis, adenomyosis or endometriosis of the abdominal wall. We performed a clinical and statistical analysis upon the collected clinical and laboratory data, together with the results obtained by using classical histological and immunohistochemical (IHC) profiling. The classical staining revealed the existence of the ectopic glandular epithelium, while the IHC reactions obtained with the anti-cytokeratin (CK) 7∕anti-CK20, anti-estrogen receptor alpha (ERα)∕anti-progesterone receptor (PR) antibodies, ascertained that these tissues were of endometrial origin. The environmental, hormonal or inflammatory factors influence these areas, so that the ER∕PR scores may be modified, the cellular proliferation might be increased (Ki67+ marker), the anti-apoptotic B-cell lymphoma 2 (BCL2) protein expression and phosphatase and tensin homolog (PTEN) may also be modified. Moreover, tumor protein 53 (p53) was positive in cases with atypia, density of inflammatory cells clearly increased compared to the adjacent normal endometrium, respectively with cluster of differentiation (CD) 3+, CD20+, CD68+, CD79a+, and tryptase+ cells, all of which may influence the cellular structure, histological architecture of the surrounding microenvironment and cause premalignant or even malignant changes in endometriosis outbreaks.

[Conjunctival stamp in diabetic patients]. / Amprenta conjunctivala la bolnavii diabetici.

UNLABELLED: The authors have suggested to reveal conjunctiva stamp changes in diabetic patients. A set of patients aged between 50 - 80 and having been diagnosed with diabetes mellitus were available for our study, therefore they have been ophthalmologically examined; both Schrimer and BUT tests have been performed. We removed conjunctiva stamps which, after being fixed and Giemsa stained, they were investigated by photonic microscopy. As to compare them we removed conjunctiva stamps from a group of control which was also Schrimer and BUT tested. RESULTS AND CONCLUSIONS: Both Schrimer and BUT tests pleaded for lacrimal hyposecretion in most of the diabetic patients; their conjunctiva stamps presented changes at the levels of both epithelial and calyciforme cells. Conjunctiva stamps were significantly changed in the old patients with a long lasted diabetes evolution pleading for the diagnosis of lacrimal hyposecretion syndrome (dry eye syndrome) in those patients.

Adenosarcoma of the uterine cervix: positive and differential diagnosis.

Adenosarcoma is considered as being a variety of mixed Müllerian tumor having a small grade of malignity. Present case refers to a perimenopause patient examined for a structure of bleeding endocervical polypoid. Diagnostic evaluation performed after biopsy revealed a cervical leiomyosarcoma followed by operation. The range of occurrence is interesting, having in mind positive and differential diagnosis problems that occurred.

The uterine carcinosarcoma - a case report.

The carcinosarcoma is a malignant mixed müllerian tumor with a highly malignant, biphasic tumor consisting of both epithelial and mesenchymal components. The presented case refers to a patient in climax with a vaginal bleeding. The Doppler echography highlights a polypoid mass, which prolapses in the cervical channel. The histopathological and immunohistochemical analysis of the surgically resected piece allowed the carcinosarcoma diagnosis. The uterine carcinosarcoma's incidence is rare, that is why this case is interesting taking in consideration the biphasic pattern of the tumor.