Molecular analysis of 31 cases with fetal skeletal dysplasia.

OBJECTIVES: The aim of this study was to describe the prenatal ultrasound findings of fetuses with skeletal dysplasia and to evaluate the genetic variations by molecular genetic analysis. METHODS: Between August 1, 2018 and March 1, 2023, we conducted a retrospective case series at a tertiary referral center involving patients with fetal skeletal abnormalities. For cases referred for a possible diagnosis of fetal skeletal dysplasia, an ultrasound database and prenatal genetic counseling records were first searched. Terminated cases diagnosed with skeletal dysplasia by pathologic and radiologic findings and cases with skeletal dysplasia proven by postnatal clinical findings were included in the study. RESULTS: Between 2018 and 2023, a total of 64 cases were diagnosed as skeletal dysplasia based on radiologic findings, pathologic findings, and clinical features. The median week of the first ultrasound performed on patients is 19 0/7 weeks, while the median week of the ultrasound in which skeletal dysplasia is suspected is 21 3/7 weeks. Although micromelia was evaluated as a common feature in all cases, the most common concomitant anomaly was thoracic hypoplasia. Exome sequencing analysis was achieved in 31 (48 %) of cases. In 31 cases, in total of 35 pathogenic single gene mutations and 5 VUS (variants of uncertain significance) variants composing of 23 autosomal dominant, 10 autosomal recessive and 2 X linked recessive mutations were determined. CONCLUSIONS: Prenatal ultrasound findings can lead us to specific diagnoses, and with the appropriate molecular analysis method, a definitive diagnosis can be made without wasting time and money.

Perspectives on carboxylates generation from Ecuadorian agro-wastes.

Carboxylates generation from banana (peel and pulp), coffee, and cacao fermentation agro-waste, upon uncontrolled and controlled pHs of 6.6 (heat-driven methanogens inactivation) and 5.2 (pH inactivation), was studied. Regarding volatile fatty acids (VFAs), acetic was the highest for cocoa (96.2 g kg-1TVS) at pH 4.5. However, butyric was relevant for banana pulp (90.7 g kg-1TVS), at controlled pH 6.6. The highest medium chain fatty acid (MCFAs) level was hexanoic (cocoa, 3.5 g kg-1TVS), while octanoic reached a maximum of 2.8 g kg-1TVS for coffee at pH 6.6. At pH 5.2 MCFAs yield was relatively low. Uncontrolled pH conditions, using banana resulted in superior VFAs production compared to controlled conditions. Thus, pH became a determining variable when deciding the time and kind of carboxylic acid to be recovered. The bacterial community at the end of the chain elongation process was dominated by phyla Firmicutes, and Clostridium as the most common genera.

Long-read next-generation sequencing for molecular diagnosis of pediatric endocrine disorders.

Recent advances in long-read next-generation sequencing (NGS) have enabled researchers to identify several pathogenic variants overlooked by short-read NGS, array-based comparative genomic hybridization, and other conventional methods. Long-read NGS is particularly useful in the detection of structural variants and repeat expansions. Furthermore, it can be used for mutation screening in difficultto- sequence regions, as well as for DNA-methylation analyses and haplotype phasing. This mini-review introduces the usefulness of long-read NGS in the molecular diagnosis of pediatric endocrine disorders.

Hepatoprotective Effect of Ethanolic Pomegranate Peel Extract Against Levofloxacin via Suppression of Oxidative Stress, Proinflammation, and Apoptosis in Male Rats.

One of the fluoroquinolone antibiotics, levofloxacin (LEV), is used to treat a variety of illnesses leading to oxidative stress and cellular damage. Peel from Punica granatum is a waste product abundant in phytochemicals with various biological activities. This study aimed to evaluate P. granatum peel extract's (PGPE) potential to mitigate oxidative stress, inflammation, apoptosis, and liver damage caused by LEV. There were four groups of rats: control, PGPE, LEV, and PGPE + LEV, respectively, and they were orally administered their daily treatments for 2 weeks. Results revealed that PGPE has a large number of phytochemical components with high antioxidant activity. PGPE intake alone enhanced the antioxidant status and decreased oxidative stress. On the other hand, pretreatment of the LEV group with PGPE restored oxidative stress, antioxidant enzymes, glutathione content, liver function biomarkers, and hematological parameters. Also, normalization of gene expressions (cyclooxygenase-2, transforming growth factor-beta1, caspase-3, heme oxygenase-1, B cell lymphoma-2, interleukin [IL]-10, and IL-1) and improvement in liver architecture, and immunohistochemical alpha-smooth muscle actin, were seen in comparison to the LEV group. Conclusively, PGPE exhibits strong anti-inflammatory, antiapoptotic, and antioxidant properties that shield rat liver from the damaging effects of LEV and offer a fresh viewpoint on the application of fruit waste products.

Asymmetric Polarization in a Rough Multilayer: Towards the Discrimination of Enantiomer Pairs.

Chirality plays a significant part in many vital processes, and to further our level of understanding, there is a steadily growing interest in enhancing the yield of enantioselective processes. Here, a multilayer with etched grooves is activated in a Kretschmann geometry and consists of alternating platinum Pt, silica SiO2, and silicon Si, as well as a silver Ag layer. Due to the production process, the groove surface exhibits a micrometric roughness, characterized by a typical vibrational mode at ω = 96 MHz. The mode is attributed to a localized acoustic vibration and has been detected as a transmitted signal. The outcomes of the inquiry include plasmonic amplification of the transmitted signal and its wavevector-less nature; in addition, it is shown that the signal is depolarized in reference to the incident beam because of the rough surface. When the Kretschmann scheme is combined with the depolarization brought on by the roughness, a built-in asymmetry results in a higher optical flux of spectrum photons in the depolarized plane than the co-polarized plane, resulting in distinct, enantioselective, and solely polarization-dependent spectral contrast. In conclusion, enantioselectivity is demonstrated for the D,L-penicillamine.

A new species of the genus Yoldiella (Bivalvia, Protobranchia, Yoldiidae) from Haima Cold Seep, South China Sea, China.

In present study, a previously unidentified but frequently encountered species of deep-sea protobranch, Yoldiellahaimaensis sp. nov., is described new to science from the Haima Cold Seep on the northwestern slope of the South China Sea. A morphological analysis confirmed that this species belongs to a previously undescribed species of the genus Yoldiella A.E. Verrill & K.J. Bush, 1897. It differs morphologically from other known species within the genus in its shell shape, degree of inflation, beaks, and number of hinge teeth. Furthermore, we sequenced three gene segments of Y.haimaensis sp. nov., comprising a nuclear ribosomal gene (18S rRNA), a nuclear protein-coding gene (histone H3), and a mitochondrial gene (cytochrome c oxidase subunit I, COI). Our phylogenetic analysis performed on the superfamily Nuculanoidea and family Yoldiidae indicates that the genus Yoldiella is non-monophyletic, and the widely recognized families within the superfamily Nuculanoidea are also not monophyletic. Our results provide molecular insights into the Protobranchia and highlight the necessity for further samples and data to revise the classification of families and genera within the superfamily using an integrative approach that combines morphological analysis and molecular data.

Acquired cystic disease associated renal cell carcinoma: A clinicopathologic and molecular study of 31 tumors.

Acquired cystic disease associated renal cell carcinomas (ACD-RCC) are rare and their molecular and histopathological characteristics are still being explored. We therefore investigated the clinicopathologic and molecular characteristics of 31 tumors. The patients were predominantly male (n = 30), with tumors mainly left-sided (n = 17), unifocal (n = 19), and unilateral (n = 29) and a mean tumor size of 25 mm (range, 3-65 mm). Microscopically, several histologic patterns were present, including pure classic sieve-like (n = 4), and varied proportions of mixed classic sieve-like with papillary (n = 23), tubulocystic (n = 9), compact tubular (n = 4) and solid (n = 1) patterns. Calcium-oxalate crystals were seen in all tumors. Molecular analysis of 9 tumors using next generation sequencing showed alterations in SMARCB1 in 3 tumors (1 with frameshift deletion and 2 with copy number loss in chromosome 22 involving SMARCB1 region), however, INI1 stain was retained in all. Nonrecurrent genetic alterations in SETD2, NF1, NOTCH4, BRCA2 and CANT1 genes were also seen. Additionally, MTOR p.Pro351Ser was identified in one tumor. Copy number analysis showed gains in chromosome 16 (n = 5), 17 (n = 2) and 8 (n = 2) as well as loss in chromosome 22 (n = 2). In summary, ACD-RCC is a recognized subtype of kidney tumors, with several histological architectural patterns. Our molecular data identifies genetic alterations in chromatin modifying genes (SMARCB1 and SETD2), which may suggest a role of such genes in ACD-RCC development.

A new species of the genus Glossobalanus (Hemichordata, Enteropneusta, Ptychoderidae) from China.

A morphological and molecular analyses of a newly discovered species, Glossobalanusweiisp. nov., from Danzhou city, Hainan Island, China is presented. Several morphological characters distinguish this new species, while molecular analyses confirm significant genetic divergence from its recognized congeners (p-distance > 0.25 in mitochondrial genomes). Phylogenetic analyses place the new species in a distinct sister clade to G.polybranchioporus, which is afforded first-class state protection in China. An updated retrieval table is provided for the eight species of Hemichordata found in China. Hemichordate diversity remains underestimated and this new species emphasizes the need for their ongoing conservation in southern China.

Kiwifruit (Actinidia deliciosa) aqueous extract improves hyperglycemia, testicular inflammation, apoptosis, and tissue structure induced by Streptozotocin via oxidative stress inhibition.

Diabetes mellitus (DM) is a well-known hyperglycemic metabolic condition identified by oxidative stress and biological function disruption. Kiwifruit is a valuable source of polyphenols and vitamin C with great antioxidant, nutritional, and health-promoting effects. Therefore, this study was initiated to explore the antioxidant and anti-hyperglycemic effects of kiwifruit aqueous extract (KFE) against oxidative injury and testis dysfunction in rats with diabetes. Twenty-four male Wistar Albino rats (160-170 g) were divided into four groups: Group 1 served as the control, Group 2 supplemented orally with kiwifruit extract (KFE; 1 g/kg/day) for one month, Group 3 was treated with a single streptozotocin dose (STZ; 50 mg/kg ip), and Group 4 where the diabetic rats were administered with KFE, respectively. According to the results, the GC-MS analysis of KFE revealed several main components with strong antioxidant properties. In diabetic rats, lipid peroxidation and hyperglycemia were accompanied by perturbations in hormone levels and sperm characteristics. Antioxidant enzymes, glutathione content, aminotransferase, phosphatase activities, and protein content were decreased. Furthermore, histology, immunohistochemical PCNA expression, and histochemical analysis of collagen, DNA, RNA, and total protein. were altered in rat testis sections, supporting the changes in biochemistry. Furthermore, diabetic rats supplemented with KFE manifested considerable amendment in all the tested parameters besides improved tissue structure and gene expressions (NF-kB, p53, IL-1ß, Bax, IL-10, and Bcl2) relative to the diabetic group. In conclusion, KFE has beneficial effects as it can improve glucose levels and testis function, so it might be used as a complementary therapy in DM.

Monitoring of toxic cyanobacterial blooms in Lalla Takerkoust reservoir by satellite imagery and microcystin transfer to surrounding farms.

Cyanobacterial harmful algal blooms (CyanoHABs) threaten public health and freshwater ecosystems worldwide. In this study, our main goal was to explore the dynamics of cyanobacterial blooms and how microcystins (MCs) move from the Lalla Takerkoust reservoir to the nearby farms. We used Landsat imagery, molecular analysis, collecting and analyzing physicochemical data, and assessing toxins using HPLC. Our investigation identified two cyanobacterial species responsible for the blooms: Microcystis sp. and Synechococcus sp. Our Microcystis strain produced three MC variants (MC-RR, MC-YR, and MC-LR), with MC-RR exhibiting the highest concentrations in dissolved and intracellular toxins. In contrast, our Synechococcus strain did not produce any detectable toxins. To validate our Normalized Difference Vegetation Index (NDVI) results, we utilized limnological data, including algal cell counts, and quantified MCs in freeze-dried Microcystis bloom samples collected from the reservoir. Our study revealed patterns and trends in cyanobacterial proliferation in the reservoir over 30 years and presented a historical map of the area of cyanobacterial infestation using the NDVI method. The study found that MC-LR accumulates near the water surface due to the buoyancy of Microcystis. The maximum concentration of MC-LR in the reservoir water was 160 µg L-1. In contrast, 4 km downstream of the reservoir, the concentration decreased by a factor of 5.39 to 29.63 µgL-1, indicating a decrease in MC-LR concentration with increasing distance from the bloom source. Similarly, the MC-YR concentration decreased by a factor of 2.98 for the same distance. Interestingly, the MC distribution varied with depth, with MC-LR dominating at the water surface and MC-YR at the reservoir outlet at a water depth of 10 m. Our findings highlight the impact of nutrient concentrations, environmental factors, and transfer processes on bloom dynamics and MC distribution. We emphasize the need for effective management strategies to minimize toxin transfer and ensure public health and safety.

Enhancing guided bone regeneration with cross-linked collagen-conjugated xenogeneic bone blocks and membrane fixation: A preclinical in vivo study.

OBJECTIVE: To determine whether combining cross-linked (CL) collagen-integrated xenogeneic bone blocks stabilized with the fixation of resorbable collagen membranes (CM) can enhance guided bone regeneration (GBR) in the overaugmented calvarial defect model. MATERIALS AND METHODS: Four circular defects with a diameter of 8 mm were prepared in the calvarium of 13 rabbits. Defects were randomly assigned to receive one of the following treatments: (i) non-cross-linked (NCL) porcine-derived collagen-embedded bone block covered by a CM without fixation (NCL + unfix group); (ii) NCL bone block covered by CM with fixation using bone-tack (NCL + fix group); (iii) cross-linked (CL) porcine-derived collagen-embedded bone block covered by CM without fixation (CL + unfix group); and (iv) CL bone block covered by CM with fixation using bone-tack fixation (CL + fix group). The efficacy of GBR was assessed through histological and molecular analyses after 2 and 8 weeks. RESULTS: At 2 weeks, there were no significant differences in histologically measured areas of newly formed bone among the groups. At 8 weeks, however, the CL + fix group exhibited a larger area of new bone (5.08 ± 1.09 mm2, mean ± standard deviation) compared to the NCL + unfix (1.62 ± 0.42 mm2; p < .0083), NCL + fix (3.97 ± 1.39 mm2) and CL + unfix (2.55 ± 1.04 mm2) groups. Additionally, the expression levels of tumour necrosis factor-alpha, fibroblast growth factor-2, vascular endothelial growth factor, osteocalcin and calcitonin receptor were significantly higher in the CL + fix group compared to the other three groups (p < .0083). CONCLUSION: Cross-linked bone blocks stabilized with collagen membrane fixation can significantly enhance GBR.

Exploring the interplay between microRNA expression and DNA mutation analysis in AML patients.

MicroRNAs (miRNAs) are key regulators in Acute Myeloid Leukemia AML, affecting gene expression, including that of CD markers and impacting mutations within leukemic cells. Mutations in AML can alter miRNA profiles, which can affect the expression of CD markers and contribute to disease progression by influencing cellular processes such as differentiation, proliferation, and apoptosis. Here, we examined the interplay of cell surface protein expression (CD markers), DNA mutations, and microRNA expression in AML patients. We included 32 recently diagnosed AML patients, and CD marker expression was evaluated using flow cytometry and molecular techniques. This study aims to delve into this relationship within the context of AML, elucidating its potential implications for diagnosis, prognosis, and therapeutic interventions. Mutations were scrutinized in six patients using Whole-Exome Sequencing (WES), while quantitative PCR (qPCR) was employed to investigate the expression levels of nine microRNAs. Subsequently, a comprehensive interaction network was constructed using Cytoscape software, focusing on genes with significant mutations and their corresponding microRNAs. Cell surface protein expression analysis revealed upregulation of CD45, CD99, CD34, HLA-DR, CD38, CD13, CD33, MPO, CD15 and CD117 in AML patients. The molecular analysis results unveiled mutations in specific genes (FLT3, KIT, PTPN11, BCR, DNMT3A, and NRAS) targeted by nine microRNAs. Notably, eight microRNAs exhibited heightened expression levels. Network analysis highlighted interactions between the PTPN11 gene and six scrutinized microRNAs. Understanding the regulatory dynamics between gene mutations and microRNAs in AML patients is pivotal for unraveling the disease's molecular mechanisms and identifying potential therapeutic targets. Further exploration into the functional roles of microRNAs in gene regulation and AML pathogenesis is warranted to validate their potential as therapeutic targets, diagnostic markers, and advanced treatment strategies.

Mass mortality of diadematoid sea urchins in the Red Sea and Western Indian Ocean.

Sea urchins are primary herbivores on coral reefs, regulating algal biomass and facilitating coral settlement and growth.1,2,3,4,5,6,7,8,9,10,11,12 Recurring mass mortality events (MMEs) of Diadema species Gray, 1825 have been recorded globally,13,14,15,16,17,18,19,20,21,22,23 the most notorious and ecologically significant of which occurred in the Caribbean in 1983,14,17,19,20 contributing to the shift from coral to algal-dominated ecosystems.17,24,25 Recently, first evidence of Diadema setosum mass mortality was reported from the eastern Mediterranean Sea.23 Here, we report extensive mass mortalities of several diadematoid species inhabiting the Red Sea and Western Indian Ocean (WIO)26,27,28 including first evidence of mortalities in the genus Echinothrix Peters, 1853. Mortalities initiated in the Gulf of Aqaba on December 2022 and span the Red Sea, the Gulf of Oman, and the Western Indian Ocean (Réunion Island), with population declines reaching 100% at some sites. Infected individuals are characterized by spine loss and tissue necrosis, resulting in exposed skeletons (i.e., tests) and mortality. Molecular diagnostics of the 18S rRNA gene confirm the presence of a waterborne scuticociliate protozoan most closely related to Philaster apodigitiformis in infected specimens-identical to the pathogen found in the 2022 Caribbean mass mortality of Diadema antillarum.13,15,18 Collapse of these key benthic grazers in the Red Sea and Western Indian Ocean may lead to algal dominance over corals, threatening the stability of coral reefs on a regional scale.29,30,31,32 We issue a warning regarding the further expansion of mortalities and call for immediate monitoring and conservation efforts for these key ecological species.

Molecular and SEM studies on Thaparocleidus vistulensis (Siwak, 1932) (Monopisthocotyla, Ancylodiscoididae).

Presenting new molecular and scanning electron microscope (SEM) features, this study gives additional data to the better knowledge of Thaparocleidus vistulensis (Siwak, 1932) (Monopisthocotyla, Ancylodiscoididae), a parasite of the European catfish Silurus glanis Linnaeus, 1758 (Siluriformes, Siluridae) cultured in a commercial fish farm in Hungary. In addition, notes on the early development of sclerotized anchors are also provided. The main morphological difference of T. vistulensis compared to other congeneric species is associated with the male copulatory organ, which exhibits 5-7 loops in the middle of the penis length and a long open V-shaped sclerotized accessory piece, dividing terminally into two parts, securing the terminal part of the penis tube. The present study provides for the first time molecular characterization data based on the 2694 bp long nucleotide sequence of rDNA (ITS1, 5.8S, ITS2, and flanked with partial 18S and partial 28S) submitted in GenBank with the accession number OR916383. A phylogenetic tree based on ITS1 sequences supports a well-defined clade including T. vistulensis, forming a sister group with T. siluri, a species-specific monopisthocotylan parasite to S. glanis. The morphological characterization of T. vistulensis, especially for the male copulatory organ, together with the molecular data in the present study, extends knowledge about this monopisthocotylan species and provides new information for future phylogeny studies.

Co-hydrothermal carbonization of municipal sludge and agricultural waste to reduce plant growth inhibition by aqueous phase products: Molecular level analysis of organic matter.

The organic matter molecular mechanism by which combined hydrothermal carbonization (co-HTC) of municipal sludge (MS) and agricultural wastes (rice husk, spent mushroom substrate, and wheat straw) reduces the inhibitory effects of aqueous phase (AP) products on pak choi (Brassica campestris L.) growth compared to HTC of MS alone is not clear. Fourier-transform ion cyclotron resonance mass spectrometry was used to characterize the differences in organic matter at the molecular level between AP from MS HTC alone (AP-MS) and AP from co-HTC of MS and agricultural waste (co-Aps). The results showed that N-bearing molecules of AP-MS and co-Aps account for 70.6 % and 54.2 %-64.1 % of all molecules, respectively. Lignins were present in the highest proportion (56.3 %-78.5 %) in all APs, followed by proteins and lipids. The dry weight of co-APs hydroponically grown pak choi was 31.6 %-47.6 % higher than that of the AP-MS. Molecules that were poorly saturated and with low aromaticity were preferentially consumed during hydroponic treatment. Molecules present before and after hydroponics were defined as resistant molecules; molecules present before hydroponics but absent after hydroponics were defined as removed molecules; and molecules absent before hydroponics but present after hydroponics were defined as produced molecules. Large lignin molecules were broken down into more unsaturated molecules, but lignins were the most commonly resistant, removed, and produced molecules. Correlation analysis revealed that N- or S-bearing molecules were phytotoxic in the AP. Tannins positively influenced the growth of pak choi. These results provide new insights into potential implementation strategies for liquid fertilizers produced from AP arising from HTC of MS and agricultural wastes.

Molecular methods confirm the presence of the alien mussel Perna viridis Linnaeus, 1758 (Bivalvia, Mytilidae) in Southern Brazil

The mussel Perna viridis, commonly known as the green mussel, is native from the Indo-Pacific region and has been introduced in various sites around the globe. In Brazil, the species has already been recorded in Rio de Janeiro and Ceará states. With the aim of assessing the presence of mussels in the southern region of the country, 14 individuals were collected in the Paranaguá Estuarine Complex, Paraná. The mussels were found attached at a depth of 2 meters on the artificial structure of Ponta do Poço Marina. The DNA was extracted using a commercial kit, the COI gene was amplified through PCR by the primers forward dgLCO-1490 and reverse dgHCO-2198, sequenced by the Sanger method, assembled in CLC Genomics Workbench, and the species was identified through the BOLD Systems. The phylogenetic tree was built on MEGA11 using 28 sequences from three species within the genus Perna. Therefore, the present study represents the first to confirm the occurrence of the exotic species Perna viridis in Brazil through molecular identification and characterizes the third Brazilian state where the mollusk has been recorded. This indicates that Brazilian coastline provides optimal environmental conditions for the establishment and development of the P. viridis

El mejillón Perna viridis, comúnmente conocido como mejillón verde, es nativo de la región del Indo-Pacífico y ha sido introducido en varios lugares alrededor del mundo. En Brasil, la especie ya ha sido registrada en los estados de Río de Janeiro y Ceará. Con el objetivo de evaluar la presencia de mejillones en la región sur del país, se recolectaron 14 individuos en el Complejo Estuarino de Paranaguá, Paraná. Los mejillones fueron encontrados fijados a una profundidad de 2 metros en la estructura artificial de la Marina Ponta do Poço. El ADN fue extraído utilizando un kit comercial, el gen COI fue amplificado mediante PCR con los cebadores forward dgLCO-1490 y reverse dgHCO-2198, secuenciados mediante el método Sanger, ensamblados en CLC Genomics Workbench, y la especie fue identificada a través de los Sistemas BOLD. El árbol filogenético fue construido en MEGA11 utilizando 28 secuencias de tres especies dentro del género Perna. Por lo tanto, el presente estudio representa el primero en confirmar la presencia de la especie exótica Perna viridis en Brasil mediante identificación molecular y caracteriza al tercer estado brasileño donde el molusco ha sido registrado. Esto indica que el litoral brasileño proporciona condiciones ambientales ideales para el establecimiento y desarrollo del P. viridis.

O mexilhão Perna viridis, comumente conhecido como mexilhão-verde, é nativo da região do Indo-Pacífico e foi introduzido em vários locais ao redor do mundo. No Brasil, a espécie já foi registrada nos estados do Rio de Janeiro e Ceará. Com o objetivo de avaliar a presença de mexilhões na região sul do país, foram coletados 14 indivíduos no Complexo Estuarino de Paranaguá, Paraná. Os mexilhões foram encontrados fixados a uma profundidade de 2 metros na estrutura artificial da Marina Ponta do Poço. O DNA foi extraído usando um kit comercial, o gene COI foi amplificado por PCR pelos primers forward dgLCO-1490 e reverse dgHCO-2198, sequenciados pelo método Sanger, montados no CLC Genomics Workbench, e a espécie foi identificada através dos Sistemas BOLD. A árvore filogenética foi construída no MEGA11 usando 28 sequências de três espécies dentro do gênero Perna. Portanto, o presente estudo representa o primeiro a confirmar a ocorrência da espécie exótica Perna viridis no Brasil através de identificação molecular e caracteriza o terceiro estado brasileiro onde o molusco foi registrado. Isso indica que o litoral brasileiro fornece condições ambientais ideais para o estabelecimento e desenvolvimento do P. viridis.

Taxonomic identification and life cycle comparison of two populations of the monostromatic green algae Monostroma nitidum.

Monostroma nitidum, a monostromatic green algae (MGA) with high economic value, is distributed worldwide. Life cycle often serves as a fundamental criterion for taxonomic classification. Most researchers consider the life cycle of M. nitidum to involve dimorphic alternation of generations, although the possibility of a monomorphic asexual life cycle remains unclear. In this study, tufA and 18S rDNA sequences were employed as molecular markers, complemented by morphological analysis, to classify and identify MGA in two distinct habitats: Hailing Island reefs (YJ) and Naozhou Island reefs (ZJ). The results of tufA and 18S rDNA sequence analysis revealed that all samples from YJ and ZJ clustered to the same branch (M. nitidum clade) with high bootstrap support and genetic distances of less than 0.000 and 0.005, respectively. However, morphological observations indicated significant differences in the external morphology of the YJ and ZJ samples, although both initially exhibited a filament-blade form during early development. The life cycle of the ZJ samples exhibited typical dimorphic alternation of generations, whereas the YJ samples only produced biflagellate asexual gametes with negative phototaxis. Gametes of the YJ samples directly developed into new gametophytes without undergoing the sporophyte stage. Consequently, the YJ and ZJ samples were classified as monomorphic asexual and dimorphic sexual M. nitidum, respectively. These findings provide evidence supporting the monomorphic asexual life cycle of M. nitidum for the classification of MGA.

Tissue Elasticity as a Diagnostic Marker of Molecular Mutations in Morphologically Heterogeneous Colorectal Cancer.

The presence of molecular mutations in colorectal cancer (CRC) is a decisive factor in selecting the most effective first-line therapy. However, molecular analysis is routinely performed only in a limited number of patients with remote metastases. We propose to use tissue stiffness as a marker of the presence of molecular mutations in CRC samples. For this purpose, we applied compression optical coherence elastography (C-OCE) to calculate stiffness values in regions corresponding to specific CRC morphological patterns (n = 54). In parallel to estimating stiffness, molecular analysis from the same zones was performed to establish their relationships. As a result, a high correlation between the presence of KRAS/NRAS/BRAF driver mutations and high stiffness values was revealed regardless of CRC morphological pattern type. Further, we proposed threshold stiffness values for label-free targeted detection of molecular alterations in CRC tissues: for KRAS, NRAS, or BRAF driver mutation-above 803 kPa (sensitivity-91%; specificity-80%; diagnostic accuracy-85%), and only for KRAS driver mutation-above 850 kPa (sensitivity-90%; specificity-88%; diagnostic accuracy-89%). To conclude, C-OCE estimation of tissue stiffness can be used as a clinical diagnostic tool for preliminary screening of genetic burden in CRC tissues.

Descriptive Study of the Clinical and Molecular Features of Epidermolysis Bullosa Patients in a Romanian European Reference Network-Skin Affiliated Reference Center.

BACKGROUND: During the last 10 years, in Romania, progress has been made for the welfare of patients suffering from epidermolysis bullosa (EB). In five university hospitals, affiliated with the National Program for the Treatment of Rare Diseases, highly trained specialists diagnose and treat patients with this rare condition. Regarding diagnosis, limitations still exist as immunofluorescence mapping and molecular genetic analysis are not accessible, and generally not reimbursed. Our objective is to present the experience in diagnosing EB patients at Colentina Clinical Hospital, highlighting genotype-phenotype correlations observed in our cohort of patients. METHODS: The records of the patients enrolled between 2012 and 2024 were analyzed considering clinical aspects, and, when available, immunofluorescence mapping, transmission electron microscopy, and genetic molecular analysis. RESULTS: Fifty-six patients were identified, of whom 31 cases were of dystrophic EB, three were of junctional EB, and 11 were of simplex EB. For 11 cases, the EB type could not be determined. Regarding EB simplex, two patients with KRT5 mutations and three patients with KRT14 mutations with various clinical expressions, from mild phenotype to severe forms, were identified. Three severe junctional EB patients were registered in our database and for one of the patients, two previously unreported mutations in the LAMA3 gene were identified. Regarding dystrophic EB, 31 cases were identified, of which 25 were recessive dystrophic cases and six were dominant dystrophic cases. Molecular genetic testing was performed for 15 patients, and the most common variant was c.425A>G, identified in six cases. DISCUSSIONS: Two previously unreported mutations were identified, namely, COL7A1 c.5416G>C, a heterozygous missense variant in a patient with a mild phenotype, mainly with nail involvement, and COL7A1 c.5960del, a variant that generates a frameshift in exon 72 resulting in a premature stop codon; this variant was identified in two siblings with a severe recessive dystrophic. CONCLUSION: Important steps have been made in identifying the correct and complete diagnosis, as well as the characterization of EB patients addressing our reference center. The findings underscore the pivotal role of molecular genetic testing in confirming diagnoses and elucidating inheritance patterns, especially in cases with atypical presentations or de novo mutations.

Molecular analysis of archival diagnostic prostate cancer biopsies identifies genomic similarities in cases with progression post-radiotherapy, and those with de novo metastatic disease.

BACKGROUND: It is important to identify molecular features that improve prostate cancer (PCa) risk stratification before radical treatment with curative intent. Molecular analysis of historical diagnostic formalin-fixed paraffin-embedded (FFPE) prostate biopsies from cohorts with post-radiotherapy (RT) long-term clinical follow-up has been limited. Utilizing parallel sequencing modalities, we performed a proof-of-principle sequencing analysis of historical diagnostic FFPE prostate biopsies. We compared patients with (i) stable PCa (sPCa) postprimary or salvage RT, (ii) progressing PCa (pPCa) post-RT, and (iii) de novo metastatic PCa (mPCa). METHODS: A cohort of 19 patients with diagnostic prostate biopsies (n = 6 sPCa, n = 5 pPCa, n = 8 mPCa) and mean 4 years 10 months follow-up (diagnosed 2009-2016) underwent nucleic acid extraction from demarcated malignancy. Samples underwent 3'RNA sequencing (3'RNAseq) (n = 19), nanoString analysis (n = 12), and Illumina 850k methylation (n = 8) sequencing. Bioinformatic analysis was performed to coherently identify differentially expressed genes and methylated genomic regions (MGRs). RESULTS: Eighteen of 19 samples provided useable 3'RNAseq data. Principal component analysis (PCA) demonstrated similar expression profiles between pPCa and mPCa cases, versus sPCa. Coherently differentially methylated probes between these groups identified ~600 differentially MGRs. The top 50 genes with increased expression in pPCa patients were associated with reduced progression-free survival post-RT (p < 0.0001) in an external cohort. CONCLUSIONS: 3'RNAseq, nanoString and 850k-methylation analyses are each achievable from historical FFPE diagnostic pretreatment prostate biopsies, unlocking the potential to utilize large cohorts of historic clinical samples. Profiling similarities between individuals with pPCa and mPCa suggests biological similarities and historical radiological staging limitations, which warrant further investigation.