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2D cell culture systems have historically provided controlled, reproducible means to analyze host-pathogen interactions observed in the human reproductive tract. Although inexpensive, straightforward, and requiring a very short time commitment, these models recapitulate neither the functionality of multilayered cell types nor the associated microbiome that occurs in a human. Animal models have commonly been used to recreate the complexity of human infections. However, extensive modifications of animal models are required to recreate interactions that resemble those in the human reproductive tract. 3D cell culture models have emerged as alternative means of reproducing vital elements of human infections at a fraction of the cost of animal models and on a scale that allows for replicative experiments. Here, we describe a new 3D model that utilizes transwells with epithelial cells seeded apically and a basolateral extracellular matrix (ECM)-like layer. The model produced tissues with morphologic and physiological resemblance to human cervical and vaginal epithelia, including mucus levels produced by cervical cells. Infection by Chlamydia trachomatis and Neisseria gonorrhoeae was demonstrated, as well as the growth of bacterial species observed in the human vaginal microbiota. This enabled controlled mechanistic analyses of the interactions between host cells, the vaginal microbiota, and STI pathogens. Affordable and semi high-throughput 3D models of the cervicovaginal epithelia that are physiologically relevant by sustaining vaginal bacterial colonization, and facilitate studies of chlamydial and gonococcal infections., (© The Author(s) 2022. Published by Oxford University Press on behalf of FEMS.)

"Leaky gut," or high intestinal barrier permeability, is common in preterm newborns. The role of the microbiota in this process remains largely uncharacterized. We employed both short- and long-read sequencing of the 16S rRNA gene and metagenomes to characterize the intestinal microbiome of a longitudinal cohort of 113 preterm infants born between 24 0/7 and 32 6/7  weeks of gestation. Enabled by enhanced taxonomic resolution, we found that a significantly increased abundance of Bifidobacterium breve and a diet rich in mother's breastmilk were associated with intestinal barrier maturation during the first week of life. We combined these factors using genome-resolved metagenomics and identified a highly specialized genetic capability of the Bifidobacterium strains to assimilate human milk oligosaccharides and host-derived glycoproteins. Our study proposes mechanistic roles of breastmilk feeding and intestinal microbial colonization in postnatal intestinal barrier maturation; these observations are critical toward advancing therapeutics to prevent and treat hyperpermeable gut-associated conditions, including necrotizing enterocolitis (NEC). IMPORTANCE Despite improvements in neonatal intensive care, necrotizing enterocolitis (NEC) remains a leading cause of morbidity and mortality. "Leaky gut," or intestinal barrier immaturity with elevated intestinal permeability, is the proximate cause of susceptibility to NEC. Early detection and intervention to prevent leaky gut in "at-risk" preterm neonates are critical for decreasing the risk of potentially life-threatening complications like NEC. However, the complex interactions between the developing gut microbial community, nutrition, and intestinal barrier function remain largely uncharacterized. In this study, we reveal the critical role of a sufficient breastmilk feeding volume and the specialized carbohydrate metabolism capability of Bifidobacterium in the coordinated postnatal improvement of the intestinal barrier. Determining the clinical and microbial biomarkers that drive the intestinal developmental disparity will inform early detection and novel therapeutic strategies to promote appropriate intestinal barrier maturation and prevent NEC and other adverse health conditions in preterm infants.

Lactobacillus iners is a common member of the human vaginal microbiota, with a genome size smaller than that of other lactobacilli. Here, we report the complete genome sequences of six L. iners strains isolated from different vaginal swab specimens. Three strains were found to harbor ∼100-kbp plasmids, which were not known previously., (Copyright © 2020 France et al.)

Bacterial vaginosis-associated bacterium 1 (BVAB1) is an as-yet uncultured bacterial species found in the human vagina that belongs to the family Lachnospiraceae within the order Clostridiales . As its name suggests, this bacterium is often associated with bacterial vaginosis (BV), a common vaginal disorder that has been shown to increase a woman's risk for HIV, Chlamydia trachomatis , and Neisseria gonorrhoeae infections as well as preterm birth. BVAB1 has been further associated with the persistence of BV following metronidazole treatment, increased vaginal inflammation, and adverse obstetrics outcomes. There is no available complete genome sequence of BVAB1, which has made it difficult to mechanistically understand its role in disease. We present here a circularized metagenome-assembled genome (cMAG) of BVAB1 as well as a comparative analysis including an additional six metagenome-assembled genomes (MAGs) of this species. These sequences were derived from cervicovaginal samples of seven separate women. The cMAG was obtained from a metagenome sequenced with long-read technology on a PacBio Sequel II instrument while the others were derived from metagenomes sequenced on the Illumina HiSeq platform. The cMAG is 1.649 Mb in size and encodes 1,578 genes. We propose to rename BVAB1 to " Candidatus Lachnocurva vaginae" based on phylogenetic analyses, and provide genomic and metabolomic evidence that this candidate species may metabolize D-lactate, produce trimethylamine (one of the chemicals responsible for BV-associated odor), and be motile. The cMAG and the six MAGs are valuable resources that will further contribute to our understanding of the heterogeneous etiology of bacterial vaginosis., (Copyright © 2020 Holm, France, Ma, McComb, Robinson, Mehta, Tallon, Brotman and Ravel.)

Functional MR imaging is being performed with increasing frequency in the typical neuroradiology practice; however, many readers of these studies have only a limited knowledge of the functional anatomy of the brain. This text will delineate the locations, anatomic boundaries, and functions of the cortical regions of the brain most commonly encountered in clinical practice-specifically, the regions involved in movement and language., (© 2019 by American Journal of Neuroradiology.)

Amplification, sequencing, and analysis of the 16S rRNA gene affords characterization of microbial community composition. As this tool has become more popular and amplicon-sequencing applications have grown in the total number of samples, growth in sample multiplexing is becoming necessary while maintaining high sequence quality and sequencing depth. Here, modifications to the Illumina HiSeq 2500 platform are described which produce greater multiplexing capabilities and 300-bp paired-end reads of higher quality than those produced by the current Illumina MiSeq platform. To improve the feasibility and flexibility of this method, a 2-step PCR amplification protocol is also described that allows for targeting of different amplicon regions, and enhances amplification success from samples with low bacterial bioburden. IMPORTANCE Amplicon sequencing has become a popular and widespread tool for surveying microbial communities. Lower overall costs associated with high-throughput sequencing have made it a widely adopted approach, especially for projects that necessitate sample multiplexing to eliminate batch effect and reduced time to acquire data. The method for amplicon sequencing on the Illumina HiSeq 2500 platform described here provides improved multiplexing capabilities while simultaneously producing greater quality sequence data and lower per-sample cost relative to those of the Illumina MiSeq platform without sacrificing amplicon length. To make this method more flexible for various amplicon-targeted regions as well as improve amplification from low-biomass samples, we also present and validate a 2-step PCR library preparation method.

Lactobacillus crispatus is a commonly found bacterium in vertebrate microbiota, particularly the human vagina. We report the first complete genome of a strain isolated from a human vagina, L. crispatus CO3MRSI1.

Vaccination is a key strategy to prevent cervical cancer in developed countries. Lower uptake of human papillomavirus (HPV) vaccine among new immigrants and refugees has been documented, although exploration of underlying reasons remains an understudied area. Semi-structured interviews with eleven immigrant women (ages 18-26 years) were conducted to understand their knowledge, attitudes and barriers regarding HPV vaccination in a western Canadian province. Participants had limited knowledge about HPV and the vaccine. Most women perceived that their risk of HPV was low, however expressed willingness to receive the vaccine if it were recommended by their physician. Greater efforts are needed to increase knowledge about HPV among immigrant and refugee women and support for physicians to discuss and offer vaccination to this underserved population.

Intestinal barrier immaturity, or "leaky gut," is the proximate cause of susceptibility to necrotizing enterocolitis in preterm neonates. However, the impact of intestinal microbiota development on intestinal mucosal barrier maturation has not been evaluated in this population. In this study, we investigated a longitudinally sampled cohort of 38 preterm infants < 33 weeks gestation monitored for intestinal permeability (IP) and fecal microbiota during the first 2 weeks of life. Rapid decrease in IP indicating intestinal barrier function maturation correlated with significant increase in community diversity. In particular, members of the Clostridiales and Bifidobacterium were highly transcriptionally active, and progressively increasing abundance in Clostridiales was significantly associated with decreased intestinal permeability. Further, neonatal factors previously identified to promote intestinal barrier maturation, including early exclusive breastmilk feeding and shorter duration antibiotic exposure, associate with the early colonization of the intestinal microbiota by members of the Clostridiales, which altogether are associated with improved intestinal barrier function in preterm infants.

Cardiac implantable electronic devices are frequently encountered in clinical practice in patients being screened for MR imaging examinations. Traditionally, the presence of these devices has been considered a contraindication to undergoing MR imaging. Growing evidence suggests that most of these patients can safely undergo an MR imaging examination if certain conditions are met. This document will review the relevant cardiac implantable electronic devices encountered in practice today, the background physics/technical factors related to scanning these devices, the multidisciplinary screening protocol used at our institution for scanning patients with implantable cardiac devices, and our experience in safely performing these examinations since 2010., (© 2017 by American Journal of Neuroradiology.)

Administration mode, age, education, and practice effects were examined for the Mental Alternation Test (MAT), a brief orally administered measure of executive function. Participants (N = 135) between the ages of 65 and 85 years completed the MAT twice in person, twice over the telephone, or once in person and once over the telephone. MAT scores did not differ across administration modes. Furthermore, the MAT detected normative decreases in executive ability in later life. The correlation between MAT performance and educational attainment was small. Finally, practice effects, which were influenced by age, were found between administration times. These findings provide important implications for both research and clinical applications of the MAT in older populations.

Background: We evaluated the utility of the telephone-administered Mental Alternation Test (MAT, an oral variant of the Trail-Making Test) for remote assessment of cognitive functioning in older adults. We examined (1) the sensitivity of MAT scores to cognitive change across 4 age groups, (2) practice effects associated with repeat administration, and (3) the uniformity of practice effects across age groups., Methods: Community-dwelling volunteers were recruited randomly and categorized as young-middle-aged (45-54 years; n = 51), middle-aged (55-64 years; n = 58), young-old (65-74 years; n = 43) or old-old (75-85 years; n = 43). The participants completed the MAT twice within 2 weeks. The data were analyzed using mixed ANOVA., Results: We found an effect of age on MAT performance [F(3, 191) = 11.37; p < 0.001], with planned comparisons revealing significantly lower scores in the old-old (p < 0.05). The scores on the second MAT administration were significantly higher than on the first administration [F(1, 191) = 12.82; p < 0.001], but this practice effect did not differ across age groups., Conclusions: The MAT was sensitive to cognitive decline in older adulthood. Practice effects were measurable but uniform across the observed age cohorts. As a brief telephone-administered test, the MAT represents a promising measure of cognitive functioning in older adults that is feasible for use in large-scale epidemiological studies., (Copyright © 2010 S. Karger AG, Basel.)

Background and Purpose: Spinal arteriovenous shunts usually require digital subtraction angiography (DSA) for evaluation. We report a unique time-resolved spinal MR angiographic (TRSMRA) technique with a temporal resolution of 3-6 seconds and spatial resolution of approximately 1 mm(3) that has the potential to noninvasively detect, localize, and follow-up these cases., Materials and Methods: Eleven patients with clinical presentation and/or MR findings suspicious for a spinal arteriovenous shunt were referred for TRSMRA. Patients subsequently underwent spinal DSA to confirm the presence or absence of a shunt or were followed clinically until an alternative diagnosis was found. TRSMRA was also used to predict the level of the shunt in the positive cases. In addition, 2 of these patients as well as a 12th patient referred to us posttreatment received a follow-up TRSMRA to assess treatment outcome., Results: Early venous shunting was identified by using TRSMRA in 6 cases. All 6 were confirmed to have an AV shunt on subsequent spinal DSA. The shunt level predicted by TRSMRA consistently correlated with DSA to within 1 vertebral level. In the 5 patients with a negative screening TRSMRA, DSA or clinical outcome confirmed the absence of an arteriovenous shunt in all of the cases. Posttreatment TRSMRA in 3 patients accurately assessed the success or failure of treatment., Conclusion: Combining acceleration techniques to achieve high frame rate TRSMRA provides sufficient temporal and spatial resolution to identify, localize, and follow patients suspected of having a spinal arteriovenous shunt. Further study in a larger population is warranted to assess the accuracy of this technique.

Elastofibroma, an unusual pseudotumor composed of excessive collagen and abnormal elastic fibers, has rarely been subjected to cytogenetic analysis. Only two cases have been previously defined, both of which demonstrated nonclonal abnormalities. In the present study, three cases of elastofibroma were cytogenetically analyzed. Abnormalities of the short arm of chromosome 1 were seen in all three cases (either clonally or as the most frequently involved region among nonclonal aberrations). In addition, a translocation involving chromosomes 8 and 12 was detected as a clonal rearrangement in one of the three cases. The observation of clonal abnormalities in elastofibroma suggests that this lesion may represent a neoplastic rather than a reactive process.

Gastrointestinal stromal tumors (GISTs), also referred to as "gastrointestinal pacemaker cell tumors (GIPACT)" are mesenchymal neoplasms that are phenotypically similar to the interstitial cells of Cajal (ICC). Cytogenetic studies of this entity are rare and molecular cytogenetic studies utilizing chromosome-specific probes are nonexistent. In the current study, cytogenetic and molecular cytogenetic analysis of 12 histologically and immunohistochemically confirmed GISTs revealed loss of a whole chromosome 14 or region(s) of 14q in 8 tumors evaluated (67%) and loss of a whole chromosome 22 or region(s) of 22q in 8 (67%) patients. Loss of 14q and 22q were observed in histologically benign and malignant GISTs. Structural rearrangements of chromosome 1 were observed in 2 malignant GISTs. These findings indicate that loss of 14q and 22q are nonrandom, early events in GIST tumorigenesis and suggest that tumor suppressor genes responsible for the development of this neoplasm may be located on these chromosomal arms.

Proliferative myositis, a reactive lesion similar to proliferative fasciitis and nodular fasciitis, has only been cytogenetically described in one other report to date. This previously described case showed trisomy 2. Cytogenetic analysis and fluorescence in situ hybridization (FISH) of a proliferative myositis lesion in the present study did not reveal trisomy 2 but the following clonal translocation was observed: 46,XX,t(6;14)(q23;q32).

Tumor-specific chromosomal abnormalities have been identified in several histologic subtypes of sarcomas. Characterization of recurrent chromosomal abnormalities has provided direction for molecular investigations of pathogenetically important genes. Cytogenetic reports of osteosarcoma, the most common primary malignant bone tumor, are relatively rare. In this study, 73 osteosarcoma specimens from 51 patients were cytogenetically analyzed following short-term culture. Clonal chromosomal abnormalities were detected in 47 and included one haploid specimen, 18 near-diploid specimens, 17 near-triploid, 8 near-tetraploid, 1 near-hexaploid, and 2 specimens with multiple clones of different ploidy levels. Examination of the present data and previously published data (111 clonally abnormal osteosarcoma specimens) reveals that chromosomal bands or regions 1p11-13, 1q10-12, 1q21-22, 11p15, 12p13, 17p12-13, 19q13, and 22q11-13 are most frequently rearranged and the most common numerical abnormalities are +1, -9, -10, -13, and -17. Partial or complete loss of the long arm of chromosome 6 also was seen in all cases of the present study and all previously published cases describing structural abnormalities of 6q. Parosteal osteosarcoma, a prognostically favorable osteosarcoma subtype, was characterized by the presence of a ring chromosome accompanied by no or few other abnormalities. Complex karyotypes were seen nearly exclusively in the high-grade lesions. These findings indicate that specific chromosomal bands and/or regions are nonrandomly involved in osteosarcoma and may provide useful clinical information.

Cytogenetic analyses of malignant peripheral nerve sheath tumors (MPNST) and malignant triton tumors (MTT) are few to date. Two separate triton tumor specimens from one patient and a MPNST specimen from another patient, both with peripheral neurofibromatosis (NF-1, von Recklinghausen disease), showed complex near-triploid complements and partial deletion of the short arm of chromosome 1. Notably, a structural abnormality of chromosome 17 was detected in the MPNST, and loss of chromosome 22 was detected in the MTT. The genes for peripheral neurofibromatosis (NF-1) and central neurofibromatosis (NF-2) have been mapped to these two chromosomes respectively.

The occurrence of telomeric associations in giant cell tumor of bone is well recognized. However, there has been little emphasis on clonal structural abnormalities that may be seen in addition to or in the absence of telomeric associations. Two cases of giant cell tumor of bone are presented in this study demonstrating clonal karyotypic anomalies other than telomeric association with a review of previously reported cases showing similar findings.

D-glycero-L-galacto-Octulose and L-glycero-L-galacto-octulose accumulated when leaves of Kenland red clover (Trifolium pratense) were allowed to imbibe solution of D-gulose or D-xylose and L-mannose or L-arabinose, respectively. The octuloses were isolated and identified by paper chromatography and by oxidative degradations to the corresponding lower sugars. Assignments of the D and L configuration were made on the basis of optical rotation. It is suggested that formation of the octuloses from the hexoses and pentoses is mediated through transketolase and aldolase or transaldolase catalysis, respectively.

1. The proteids consisted of albumins and globulins, varying in quantity between the extremes of .9010 grm. and .2592 grm. in the 24 hours, but usually between .7 and .37 grm., the average being .5317 grm. 2. The quantity of the proteids varies directly as the urea, inversely as the external temperature. The relation between the quantity of urine and the quantity of proteid is not constant. The quantity of the proteida is little affected by diuretics; it tends to be increased by certain drugs that act upon the circulation. It is lessened during sleep. It is apt to suffer a sudden temporary increase, returning as suddenly to the usual average. For this phenomenon we have no explanation. 3. From .5449 to .6616 grm. of coagulable proteids a day (.0079 to .0094 grm. per kilo of body-weight) may be excreted through the urine for an indefinite time by an otherwise healthy individual without damage to either kidneys or organism.

Experimental upper respiratory infections similar to "common colds" were transmitted singly and in series through two and four passages in nine out of fifteen persons, by intransal inoculations with bacteria-free filtrates of nasopharyngeal washings obtained from individuals ill with natural "colds." These observations conform with those reported by previous workers and lend further support to the view that the incitant of the "common cold" is a filtrable virus.