Prenatal exome sequencing and chromosomal microarray analysis in fetal structural anomalies in a highly consanguineous population reveals a propensity of ciliopathy genes causing multisystem phenotypes.

Fetal abnormalities are detected in 3% of all pregnancies and are responsible for approximately 20% of all perinatal deaths. Chromosomal microarray analysis (CMA) and exome sequencing (ES) are widely used in prenatal settings for molecular genetic diagnostics with variable diagnostic yields. In this study, we aimed to determine the diagnostic yield of trio-ES in detecting the cause of fetal abnormalities within a highly consanguineous population. In families with a history of congenital anomalies, a total of 119 fetuses with structural anomalies were recruited and DNA from invasive samples were used together with parental DNA samples for trio-ES and CMA. Data were analysed to determine possible underlying genetic disorders associated with observed fetal phenotypes. The cohort had a known consanguinity of 81%. Trio-ES led to diagnostic molecular genetic findings in 59 fetuses (with pathogenic/likely pathogenic variants) most with multisystem or renal abnormalities. CMA detected chromosomal abnormalities compatible with the fetal phenotype in another 7 cases. Monogenic ciliopathy disorders with an autosomal recessive inheritance were the predominant cause of multisystem fetal anomalies (24/59 cases, 40.7%) with loss of function variants representing the vast majority of molecular genetic abnormalities. Heterozygous de novo pathogenic variants were found in four fetuses. A total of 23 novel variants predicted to be associated with the phenotype were detected. Prenatal trio-ES and CMA detected likely causative molecular genetic defects in a total of 55% of families with fetal anomalies confirming the diagnostic utility of trio-ES and CMA as first-line genetic test in the prenatal diagnosis of multisystem fetal anomalies including ciliopathy syndromes.

Lessons Learned from Large-Scale, First-Tier Clinical Exome Sequencing in a Highly Consanguineous Population.

We report the results of clinical exome sequencing (CES) on >2,200 previously unpublished Saudi families as a first-tier test. The predominance of autosomal-recessive causes allowed us to make several key observations. We highlight 155 genes that we propose to be recessive, disease-related candidates. We report additional mutational events in 64 previously reported candidates (40 recessive), and these events support their candidacy. We report recessive forms of genes that were previously associated only with dominant disorders and that have phenotypes ranging from consistent with to conspicuously distinct from the known dominant phenotypes. We also report homozygous loss-of-function events that can inform the genetics of complex diseases. We were also able to deduce the likely causal variant in most couples who presented after the loss of one or more children, but we lack samples from those children. Although a similar pattern of mostly recessive causes was observed in the prenatal setting, the higher proportion of loss-of-function events in these cases was notable. The allelic series presented by the wealth of recessive variants greatly expanded the phenotypic expression of the respective genes. We also make important observations about dominant disorders; these observations include the pattern of de novo variants, the identification of 74 candidate dominant, disease-related genes, and the potential confirmation of 21 previously reported candidates. Finally, we describe the influence of a predominantly autosomal-recessive landscape on the clinical utility of rapid sequencing (Flash Exome). Our cohort's genotypic and phenotypic data represent a unique resource that can contribute to improved variant interpretation through data sharing.

Primary and Secondary Breast Lymphoma: Clinical, Pathologic, and Multimodality Imaging Review.

Breast lymphoma is a rare hematologic neoplasm that originates in the breast lymphoid tissue and includes primary breast lymphoma (PBL) and secondary breast lymphoma (SBL). PBL involves the breast lymphoid tissue in the absence of previously identified extramammary lymphoma and widespread disease. SBL is the most common metastasis to the breast, accounting for 17% of metastatic disease to the breast. PBL and SBL usually demonstrate imaging phenotypes that overlap with those of primary breast carcinoma, which makes a prospective diagnosis of breast lymphoma challenging. These nonspecific imaging features include an iso- to hyperdense oval mass or masses at mammography, a hypoechoic or mixed-echogenicity hypervascular mass at US, an enhancing mass with type II kinetics at MRI, and high fluorine 18-fluorodeoxyglucose avidity at PET. In cases of suspected lymphoma, reviewing the clinical history, using appropriate biopsy techniques, and evaluating for multiplicity, bilaterality, and distant disease are critical for diagnosis and management. A patient with PBL generally has an earlier clinical presentation with a palpable abnormality and a solitary imaging finding. In contrast, multiple masses in an older patient and an occult clinical presentation favor an SBL diagnosis. ©RSNA, 2019.

Incorporation of zinc sulfide nanoparticles, Acinetobacter pittii and Bacillus velezensis to improve tomato plant growth, biochemical attributes and resistance against Rhizoctoniasolani.

Green nanobiotechnology and beneficial bacterial strains as biofertilizers are crucial in agriculture to achieve food security. Both these strategies have been individually studied in improving plant resistance against phytopathogens along with enhancing plant productivity. Therefore, objective of this study was to explore the eco-friendly and cost-effective approach of utilizing plant growth promoting and disease suppressing bacterial strains and nanoparticles, individually as well as in combination, as bio-stimulants to improve plant growth, antioxidant defense system, nutrition and yield of tomato. A pot experiment was conducted to investigate the zinc sulfide nanoparticles (ZnS NPs) synthesized by using Jacaranda mimosifolia flower extracts (JFE), Acinetobacter pittii and Bacillus velezensis either individually or in combinations to check their potential against Rhizoctonia solani in tomato to suppress root rot infection and improve growth and yield. Among all the combinations the JFE-ZnS NPs + B. velezensis compared to untreated infected plants showed minimum disease incidence and maximum significant protection (66%) against R. solani instigated root rot that was followed by JFE-ZnS NPs + A. pittii and individual application of JFE-ZnS NPs by 58%. The same treatment showed maximum significant increase in plant fresh and dry biomass. B. velezensis significantly increased the photosynthetic pigments when applied individually. However, JFE-ZnS NPs alone and in mixed treatments with B. velezensis efficiently improved total soluble protein, sugar and phenolic contents. The same interactive application of JFE-ZnS NPs + B. velezensis improved the tomato plant nutrition (silicon (Si), magnesium (Mg), calcium (Ca) and potassium (K)) and redox quenching status by improving the activity of antioxidant defense enzymes. Overall, the interactive use of JFE-ZnS NPs with A. pittii and B. velezensis very appropriately prepared the host plant to fight against the negative effects of root rot pathogen in tomato. Advancements in interactively investigating the nanoparticles with beneficial plant growth promoting bacterial strains importantly can contribute in resolving the challenges of food security. According to our information, this is a pioneer report for implying JFE-ZnS NPs in synergism with A. pittii and B. velezensis to hinder the root rot in tomatoes.

Clinico-radiologic features and management of hematological tumors in the breast: a case series.

Hematological tumors arising in the breast are uncommon and require different treatment modalities dependent upon tumor type. Current treatment options include surgical excision, chemotherapy, and radiotherapy. Management of these breast malignancies are poorly outlined in the literature. The purpose of this case series is to report five cases consisting of extranodal marginal zone lymphoma, lymphoplasmacytic lymphoma, and extramedullary plasmacytoma occurring in the breast. The cases illustrate heterogeneous radiologic findings and varying management approaches to these tumors. The case series underscores the importance of having a wide differential at diagnosis and recognizes management of disease should be taken on an individual basis with consideration of prognosis and first-line treatment options.

Novel CARMIL2 Mutations in Patients with Variable Clinical Dermatitis, Infections, and Combined Immunodeficiency.

Combined immunodeficiencies are a heterogeneous collection of primary immune disorders that exhibit defects in T cell development or function, along with impaired B cell activity even in light of normal B cell maturation. CARMIL2 (RLTPR) is a protein involved in cytoskeletal organization and cell migration, which also plays a role in CD28 co-signaling of T cells. Mutations in this protein have recently been reported to cause a novel primary immunodeficiency disorder with variable phenotypic presentations. Here, we describe seven patients from three unrelated, consanguineous multiplex families that presented with dermatitis, esophagitis, and recurrent skin and chest infections with evidence of combined immunodeficiency. Through the use of whole exome sequencing and autozygome-guided analysis, we uncovered two mutations not previously reported (p.R50T and p.L846Sfs) in CARMIL2. Real-time PCR analysis revealed that the biallelic frameshift mutation is under negative selection, likely due to nonsense-mediated RNA decay and leading to loss of detectable protein upon immunoblotting. Protein loss was also observed for the missense mutation, and 3D modeling suggested a disturbance in structural stability due to an increase in the electrostatic energy for the affected amino acid and surrounding residues. Immunophenotyping revealed that patient Treg counts were significantly depressed, and that CD4+ T cells were heavily skewed towards the naïve status. CD3/CD28 signaling impairment was evidenced by reduced proliferative response to stimulation. This work broadens the allelic heterogeneity associated with CARMIL2 and highlights a deleterious missense alteration located outside the leucine-rich repeat of the protein, where all other missense mutations have been reported to date.

Diagnosis of sarcoidosis from a biopsy of a dilated mammary duct.

Sarcoidosis is an immunologic disease of unknown etiology that manifests most frequently within the lungs or associated lymph nodes. Sarcoidosis involving the breast is seen in <1% of cases and usually is diagnosed in patients with multisystem disease. The clinical and imaging presentations of sarcoidosis of the breast can be variable. Though uncommon, sarcoidosis should be considered in the differential diagnosis of a breast lesion, and given that imaging characteristics cannot distinguish between sarcoidosis and malignancy, all breast lesions in patients with sarcoidosis should be biopsied. Our case study demonstrates a diagnosis of sarcoidosis in an asymptomatic patient presenting with a single dilated duct and associated filling defect within the right breast.

Comparative analysis of bioactive N-alkylamides produced by tissue culture raised versus field plantlets of Spilanthes ciliata using LC-Q-TOF (HRMS).

Spilanthes ciliata (S. ciliata) is a perennial herb of global importance owing to its luscious source of bioactive fatty acid derived amides known as N-alkylamides. It finds application in skin creams, mouth gels and toothpastes. Despite multifaceted applications, a major limitation associated for its commercial application is the scarcity of contamination free plant source, fluctuations in active metabolites due to variation in extraction procedures, and lack of rapid qualitative method for alkylamide profiling. In the current work, attempts were made to 1) optimize conditions for mass propagation of contamination free plants of S. ciliata by tissue culture using leaf discs as explants, 2) establish an optimum extraction ratio of plant/solvent (w/v) for maximum elution of alkylamides and 3) develop a rapid method for qualitative estimation of alkylamide from in vitro raised plants in comparison with that of the field grown counterpart by using LC-Q-TOF (HRMS). To the best of our knowledge, this is the first qualitative report on alkylamide profile of micropropagated whole plant of Spilanthes. The correlation pattern reported in this study may form the basis for using tissue culture raised plantlets of S. ciliata as potential source of bioactive alkylamides on industrial scale.

Isolation, characterization and HPLC quantification of compounds from Aquilegia fragrans Benth: Their in vitro antibacterial activities against bovine mastitis pathogens.

ETHNO-PHARMACOLOGICAL RELEVANCE: The underground parts of Aquilegia fragrans are traditionally used for the treatment of wounds and various inflammatory diseases like bovine mastitis. However, there are no reports on the phytochemical characterization and antibacterial studies of A. fragrans. AIM OF THE STUDY: To isolate compounds from the methanol extract of the underground parts of A. fragrans and determine their antibacterial activity against the pathogens of bovine mastitis. The study was undertaken in order to scientifically validate the traditional use of A. fragrans. MATERIALS AND METHODS: Five compounds were isolated from the methanol extract of the underground parts of A. fragrans using silica gel column chromatography. Structural elucidation of the isolated compounds was done using spectral data analysis and comparison with literature. High performance liquid chromatography (HPLC) was used for the qualitative and quantitative determination of isolated compounds in the crude methanol extract. The methanol extract and isolated compounds were evaluated for antibacterial activities against mastitis pathogens using broth micro-dilution technique. RESULTS: The five isolated compounds were identified as (1) 2, 4-dihydroxyphenylacetic acid methyl ester (2) ß-sitosterol (3) Aquilegiolide (4) Glochidionolactone-A and (5) Magnoflorine. A quick and sensitive HPLC method was developed for the first time for qualitative and quantitative determination of four isolated marker compounds from A. fragrans. The crude methanol extract and compound 5 exhibited weak antibacterial activities that varied between the bacterial species (MIC=500-3000 µg/ml). CONCLUSIONS: The above results show that the crude methanol extract and isolated compounds from A. fragrans exhibit weak antibacterial activities. Further phytochemical and pharmacological studies are required for proper scientific validation of the folk use of this plant species in the treatment of various inflammatory diseases like bovine mastitis.

Impact of Henda's law on the utilization of screening breast magnetic resonance imaging.

Female breast tissue is composed of variable proportions of fat and fibroglandular tissue, and in general, an increased ratio of fibroglandular tissue to fat corresponds to increased mammographic density. Studies suggest that mammographic density is an independent risk factor for breast cancer, and the sensitivity of mammography can be lower with heterogeneously dense or extremely dense breasts. Nineteen states have legal statutes requiring that patients be notified if they have dense breasts, including the state of Texas. Henda's law, mandated on January 1, 2012 in Texas, suggests that patients with dense breasts could benefit from additional screening tests such as breast magnetic resonance imaging (MRI). Our study examined the impact of Henda's law by comparing the number of screening breast MRIs performed for dense breasts before and after the law's implementation. Results showed a 23-fold increase in the number of dense breast MRIs in the 2 years that this new legislation was in effect. This increase could have substantial implications for the health care economy, and further studies are needed to determine the cost-effectiveness of this additional screening tool.

Unusual presentation of metastatic ovarian carcinoma as an enlarged intramammary lymph node.

Metastasis to the breast most commonly arises from a contralateral primary breast malignancy; however, metastatic disease can also result from extramammary malignancies by hematogenous or lymphatic dissemination. This case report reviews an unusual presentation of primary ovarian carcinoma with metastasis to an intramammary lymph node.

Clinical utility of positron emission mammography.

Several imaging modalities have been introduced over recent years to better screen for and stage breast cancer. Positron emission mammography (PEM) has been approved by the US Food and Drug Administration and introduced into clinical use as a diagnostic adjunct to mammography and breast ultrasonography. PEM has higher resolution and a more localized field of view than positron emission tomography-computed tomography and can be performed on patients to stage a newly diagnosed malignancy. Review of mammograms together with magnetic resonance or PEM images improves detection of disease.

Percutaneous tattoo pigment simulating calcific deposits in axillary lymph nodes.

The isolated finding of calcific deposits within axillary lymph nodes on mammography suggests a broad range of differential diagnoses, from benign causes such as granulomatous reaction secondary to previous histoplasmosis infection to malignancies such as breast cancer and metastatic disease from extramammary primary malignancies. Therefore, the isolated finding of intranodal calcium may warrant biopsy for a definitive diagnosis when a benign etiology is not apparent. We present a patient with isolated axillary lymph node densities on mammography and chest computed tomography, which were subsequently proven to represent deposition of tattoo pigment.

The radial scar of the breast diagnosed at core needle biopsy.

The radial scar (RS) or complex sclerosing lesion (CSL) of the breast represents a management dilemma on diagnosis at breast core needle biopsy because of the risk of associated malignancy identified only upon surgical excision. To determine our experience, we retrospectively reviewed core needle biopsies performed at the Darlene G. Cass Breast Imaging Center from 2006 to 2011, identifying 67 patients with RS or CSL, and correlated histology at excisional biopsy with core biopsy results. Of the 67 cases, 6 (9%) were associated with malignancy at surgical excision. The average size of the RS or CSL was 1.42 cm. In conclusion, RS or CSL diagnosed at core needle biopsy still warrants surgical excision because of the significant percentage (9%) of cases with associated malignancy.