Maintenance of bacterial outer membrane lipid asymmetry: insight into MlaA.

The outer membrane (OM) of Gram-negative bacteria acts as an effective barrier to protect against toxic compounds. By nature, the OM is asymmetric with the highly packed lipopolysaccharide (LPS) at the outer leaflet and glycerophospholipids at the inner leaflet. OM asymmetry is maintained by the Mla system, in which is responsible for the retrograde transport of glycerophospholipids from the OM to the inner membrane. This system is comprised of six Mla proteins, including MlaA, an OM lipoprotein involved in the removal of glycerophospholipids that are mis-localized at the outer leaflet of the OM. Interestingly, MlaA was initially identified - and called VacJ - based on its role in the intracellular spreading of Shigella flexneri.Many open questions remain with respect to the Mla system and the mechanism involved in the translocation of mislocated glycerophospholipids at the outer leaflet of the OM, by MlaA. After summarizing the current knowledge on MlaA, we focus on the impact of mlaA deletion on OM lipid composition and biophysical properties of the OM. How changes in OM lipid composition and biophysical properties can impact the generation of membrane vesicles and membrane permeability is discussed. Finally, we explore whether and how MlaA might be a candidate for improving the activity of antibiotics and as a vaccine candidate.Efforts dedicated to understanding the relationship between the OM lipid composition and the mechanical strength of the bacterial envelope and, in turn, how such properties act against external stress, are needed for the design of new targets or drugs for Gram-negative infections.

Predictors of disease severity in SARS-CoV-2 omicron variant XBB sublineages and variants of interest.

The Omicron variant has been reported to present with milder disease compared with Delta, although this may be due to immunity from vaccination and prior exposure. Predictors of severity with recent strains have not been well characterized. We retrospectively examined consecutive cases of moderate-to-severe COVID-19 (defined as requiring supplemental oxygenation, intensive care or mortality) admitted to seven tertiary hospitals across Singapore in April 2023. Whole genome sequencing was performed on each isolate to determine the sublineage, while baseline clinical, laboratory data and outcomes were tabulated. We reviewed 182 patients with moderate-to-severe illness and 466 controls hospitalized at the same time. Advanced age and presence of chronic kidney disease predicted adverse outcome. Previously reported markers such as radiographic evidence of pneumonia, elevated C-reactive protein and serum creatinine levels at presentation also correlated with adverse outcomes. There were no observable differences in outcomes with any specific Omicron XBB sublineage. We did not find any specific Omicron XBB sublineage that was associated with worse outcomes. Larger multinational studies would be important to track the clinical evolution of the virus in its current endemic state.

Cognitive performance of women at various stages of reproductive aging and associated risk factors.

OBJECTIVE: Menopausal transition among midlife women is accompanied by the issues of hot flushes, mood swings as well as sleep disturbances, night sweats, urogenital diminution and cognitive changes. The present cross-sectional study is an attempt to recognize the cognitive differences associated with various stages of reproductive aging. METHODS: The study included women (N = 404) aged between 40 and 65 years from rural areas of Punjab (North India). The Mini-Mental State Examination (Hindi version) was used to evaluate orientation, registration, attention, recall, language and visual spatial skills domains of the global cognitive functioning of all participants. RESULTS: The results of the present study exhibited a decline in cognitive scores across successive menopausal transitional stages in most of the cognitive domains except registration. The results of multivariate regression analysis demonstrated that illiteracy, vegetarian diet, perimenopause and late postmenopause phase were significantly associated with lower global cognitive scores. CONCLUSION: With increased longevity, early identification of potential risk factors associated with cognitive decline among women during their midlife can be beneficial in improving the mental health of postmenopausal women.

Frontal fibrosing alopecia: survey of severity assessment methods in routine clinical practice and validation of the International Frontal Fibrosing Alopecia Cooperative Group measurement guidance.

BACKGROUND: The lack of validated and responsive outcome measures in the management of frontal fibrosing alopecia (FFA) significantly limits assessment of disease progression and treatment response over time. AIM: To understand how FFA extent and progression is currently assessed in UK specialist centres, to validate components of the International FFA Cooperative Group (IFFACG) statement on FFA assessment, and to identify pragmatic advice to improve FFA management in clinic. METHODS: Consultant dermatologists with a specialist interest in hair loss (n = 17) were invited to take part. Preferred FFA assessment methods were explored using questionnaires and clinical scenarios. Participants were asked to identify and mark the current hairline in 10 frontal and 10 temporal hairline images (Questionnaire 1), with assessment repeated 3 months later to assess intraindividual variability (Questionnaire 2) and 12 months later to test whether interindividual accuracy could be improved with simple instruction (Questionnaire 3). RESULTS: All 17 clinicians (100%) completed the questionnaire at each time interval. We identified a wide variation in assessment techniques used by our experts. Measurements were perceived as the most accurate method of assessing frontal recession whereas photography was preferred for temporal recession. Inter-rater reliability between clinicians measuring the frontal hairline scenarios indicated a moderate strength of agreement [intraclass coefficient (ICC) = 0.61; 95% CI 0.40-0.85], yet intrarater reliability was found to be poor with wide limits of agreement (-8.71 mm to 9.92 mm) on follow-up. Importantly, when clear guidance was provided on how the hairline should be identified (Questionnaire 3), inter-rater reliability improved significantly, with ICC = 0.70, suggesting moderate agreement (95% CI 0.51-0.89; P < 0.001). A similar pattern was seen with temporal hairline measurements, which again improved in accuracy with instruction. CONCLUSION: We found that accuracy of measurements in FFA can be improved with simple instruction and we have validated components of the IFFACG measurement recommendations.

Post-trabeculectomy early bleb localization mimicking a 'ring of steel'.

A 54-year-old-man underwent trabeculectomy with mitomycin-C (0.02%). At 4-week follow-up, there was an elevated, well-circumscribed bleb with raised intra-ocular pressure (IOP) of 32-mmHg, with a classic "ring of steel" appearance, typically seen in a tenon's cyst. A sub-tenon needle revision re-established aqueous flow. Two weeks later, the IOP was 8-mmHg without any anti-glaucoma medications. A diffuse well formed bleb was noted. Careful ocular examination helps to identify early post-operative localized bleb that may appear like a "ring of steel" but can be managed by needling, unlike a tenon's cyst, which would require more extensive intervention.

Infant precursors of executive function in Down syndrome.

BACKGROUND: Although early features of infant cognition are predictive of executive function (EF) in typically developing (TD) children, there is little information regarding the developmental origins of EF in neurogenetic conditions, such as Down syndrome (DS). METHODS: The current study compared the performance of infants with and without DS on three dimensions that are hypothesised EF precursors: visual engagement, attention shifting and action planning. Additionally, the relationship between these EF precursors at Time 1 and EF performance at Time 2 (6 months later) was examined in the DS group. Participants were 36 infants with DS, M chronological age = 12.65 months, SD = 2.11; M developmental age = 8.84 months, SD = 2.22, and 36 TD infants, M chronological age = 8.62, SD = 3.06; M developmental age = 8.64 months, SD = 3.40. RESULTS: Infants with DS visually engaged with objects for longer durations and demonstrated challenges with action planning compared with TD infants at Time 1. Attention shifting at Time 1 significantly predicted EF performance at Time 2 in the DS group. CONCLUSIONS: This study provides evidence that an early atypical presentation of EF precursors is detectable during infancy in DS and is predictive of subsequent EF performance. These findings contribute to the identification of areas of early cognitive risk in DS and can inform future interventions in this population.

Duodenal eosinophilia is associated with symptomatic erosive gastro-oesophageal reflux disease, presence of co-morbidities, and ethnicity but not undifferentiated functional dyspepsia: A retrospective Malaysian study.

BACKGROUND: Duodenal eosinophilia is postulated to play a key role in the pathogenesis of functional dyspepsia, a common condition responsible for considerable impairment of quality of life. Our objective was to evaluate the relative strength of the associations between duodenal eosinophilia, functional dyspepsia, symptomatic erosive gastroesophageal reflux disease (GERD), the presence of co-morbidities, and a number of other variables. METHODS: Eosinophil counts of archived endoscopic duodenal biopsies of 289 subjects were determined by a pathologist blinded to the clinical data. Duodenal eosinophilia was defined by a count of more than 15 per 5 high power fields. Clinical charts were reviewed by a gastroenterologist blinded to the histology review. RESULTS: In the study sample, the primary diagnosis was functional dyspepsia (undifferentiated by subtypes) in 45, symptomatic erosive GERD in 29, gall stone disease in 17, irritable bowel syndrome in 23, and an alternative or undetermined diagnosis in 175 subjects, respectively. On logistic regression analyses, eosinophil counts were positively associated with symptomatic erosive GERD (Odds Ratio, OR 1.03, 95% Confidence Interval, 95%CI: 1.00, 1.05; p=0.035) but not functional dyspepsia. Pre-defined duodenal eosinophilia was associated with symptomatic erosive gastro-oesophageal reflux disease (OR 3.36, 95%CI 1.18,-9.60; p=0.023), the presence of co-morbidities (OR 2.00, 95%CI 1.10, 3.62; p=0.022), and Chinese (as compared to Malay and Indian) ethnicity but not with either functional dyspepsia, irritable bowel syndrome, gallstone disease, Helicobacter pylori infection, or gender. CONCLUSION: Duodenal eosinophilia was associated with symptomatic erosive GERD, the presence of co-morbidities, and Chinese ethnicity but not with undifferentiated functional dyspepsia.

Recent developments in sustainably sourced protein-based biomaterials.

Research into the development of sustainable biomaterials is increasing in both interest and global importance due to the increasing demand for materials with decreased environmental impact. This research field utilises natural, renewable resources to develop innovative biomaterials. The development of sustainable biomaterials encompasses the entire material life cycle, from desirable traits, and environmental impact from production through to recycling or disposal. The main objective of this review is to provide a comprehensive definition of sustainable biomaterials and to give an overview of the use of natural proteins in biomaterial development. Proteins such as collagen, gelatin, keratin, and silk, are biocompatible, biodegradable, and may form materials with varying properties. Proteins, therefore, provide an intriguing source of biomaterials for numerous applications, including additive manufacturing, nanotechnology, and tissue engineering. We give an insight into current research and future directions in each of these areas, to expand knowledge on the capabilities of sustainably sourced proteins as advanced biomaterials.

Denoising AVIRIS-NG Data for Generation of New Chlorophyll Indices.

The availability of Airborne Visible and Infrared Imaging Spectrometer-Next Generation (AVIRIS-NG) data has enormous possibilities for quantification of Leaf Chlorophyll Content (LCC). The present study used the AVIRIS-NG campaign site of Western India for generation and validation of new chlorophyll indices by denoising the AVIRIS-NG data. For validation, concurrent to AVIRIS-NG flight overpass, field samplings were performed. The acquired AVIRIS-NG was subjected to Spectral Angle Mapper (SAM) classifier for discriminating the crop types. Three smoothing techniques i.e., Fast-Fourier Transform (FFT), Mean and Savitzky-Golay filters were evaluated for their denoising capability. Raw and filtered data was used for developing new chlorophyll indices by optimizing AVIRIS-NG bands using VIs based on parametric regression algorithms. In total, 20 chlorophyll indices and corresponding 20 models were developed for mapping LCC in the area. SAM identified 17 crop types in the area, while FFT found to be the best for filtering. Performance of these models when checked based on Pearson correlation coefficient (r) and Centered Root Mean Square Difference (CRMSD), indicated that LCC-CCI10 based on normalized difference type index formed through Near Infrared band and blue band is the best estimator of LCC (rcal = 0.73, rval = 0.66, CRMSD = 4.97). The approach was also tested using AVIRIS-NG image of the year 2018, which also showed a promising correlation (r = 0.704, CRSMD = 8.98, Bias = -0.5) between modeled and field LCC.

An endophytic Penicillium oxalicum isolated from Citrus limon possesses antioxidant and genoprotective potential.

AIMS: This study aimed at isolating endophytic fungi from Citrus limon (L.) possessing antioxidative and genoprotective potential. METHODS AND RESULTS: Endophytic fungi were screened for antioxidant activity using 2,2-diphenyl,1-picryl hydrazyl radical scavenging assay and maximum activity (79·70%) was exhibited by culture MP1 identified to be Penicillium oxalicum on the basis of morphological and molecular characteristics. The ethyl acetate extract of MP1 was subjected to silica column chromatography followed by LH 20 column chromatography for purification of active metabolites. The partially purified active fraction of P. oxalicum MP1 possessed good antioxidant activity as detected using various assays. It also exhibited a strong DNA damage protection potential on pUC19 plasmid DNA treated with Fenton reagent. On exposure to active fraction of MP1 significant reduction (P < 0·05) in nuclear deformities (like nuclear buds, micronuclei, nuclear ridges and binucleated cells) was observed in human lymphocytes pretreated with a toxic concentration of H2 O2 . In vivo genoprotectivity studies were conducted in fresh water fish Channa punctatus pretreated with a damaging compound 4-nonyl phenol. The active fraction of P. oxalicum MP1 caused a reduction of 94·7 and 66·60% in micronuclei and aberrant cell formation, respectively. A significant reduction (P < 0·05) in tail length and tail DNA parameters was also observed in comet assay. CONCLUSION: The endophytic P. oxalicum isolated in this study has the potential to produce metabolites possessing antioxidant and genoprotective activities. SIGNIFICANCE AND IMPACT OF THE STUDY: The isolated culture can be exploited in the field of therapeutics by virtue of its in vitro and in vivo genoprotective potential.

Goal-directed action planning in infants with Down syndrome.

BACKGROUND: Down syndrome (DS) is a neurogenetic disorder associated with risk for executive dysfunction, or difficulties with the cognitive processes required for planning volitional, goal-directed behaviour. This study examines the developmental origins of difficulties with goal-directed action planning in infants with DS to inform our understanding of the cognitive phenotype associated with DS and its implications for intervention. METHOD: First, the study compared the performance of infants with DS (n = 44, mean chronological age = 7.5 months, SD = 2.3) and typically developing infants (n = 31, mean chronological age = 7.5 months, SD = 2.9) on plan production and planning efficiency during an early planning task. Next, potential sources of variability in planning behaviour (motor performance and sensory processing) within the DS sample were examined. All infants completed an early planning laboratory task and the Bayley Scales of Infant Development-III Cognitive Scale. The motor and sensory skills of infants with DS were assessed by the motor scales of the Bayley Scales of Infant Development-III and the Infant Sensory Profile-2. DS-related biomedical history information was provided by caregivers for the infants with DS. RESULTS: Between-group differences in planning were observed on the dimensions of strategy production and completion, such that infants with DS were less efficient in their strategy execution than typically developing infants. In the DS group, motor skills and sensory processing were associated with planning efficiency on all components of the early planning task. CONCLUSIONS: Less efficient action planning in infants with DS may disrupt the shaping of goal-directed behaviour, and the identification of early risk factors associated with planning efficiency has important implications for early intervention.

Benefit-risk profile of tofacitinib in patients with moderate-to-severe chronic plaque psoriasis: pooled analysis across six clinical trials.

BACKGROUND: Although existing psoriasis treatments are effective and well tolerated in many patients, there is still a need for new effective targeted treatment options. Tofacitinib is an oral Janus kinase inhibitor that has been investigated in patients with moderate-to-severe chronic plaque psoriasis. OBJECTIVES: To consider the benefits and risks of tofacitinib in patients with moderate-to-severe psoriasis. METHODS: Data were pooled from one phase II, four phase III and one long-term extension study comprising 5204 patient-years of tofacitinib treatment. Efficacy end points included patients achieving Physician's Global Assessments of 'clear' or 'almost clear', ≥ 75% and ≥ 90% reduction in Psoriasis Area and Severity Index (coprimary end points) and improvements in Dermatology Life Quality Index score, Hospital Anxiety and Depression Scale depression score and Itch Severity Item score, at weeks 16 and 52. Safety data were summarized for 3 years of tofacitinib exposure. RESULTS: Tofacitinib 5 and 10 mg twice daily (BID) showed superiority over placebo for all efficacy end points at week 16, with response maintained for 52 weeks of continued treatment. Tofacitinib improved patients' quality of life and was well tolerated. Rates of safety events of interest (except herpes zoster) were similar to those in the published literature and healthcare databases for other systemic psoriasis therapies. Tofacitinib 10 mg BID demonstrated greater efficacy than 5 mg BID. CONCLUSIONS: Tofacitinib has a benefit-risk profile in moderate-to-severe psoriasis consistent with that of other systemic treatments.

Evaluation of genotoxic and cytotoxic effects of ethyl acetate extract of Aspergillus flavus on Spodoptera litura.

AIM: Recent concerns about the possible adverse effects of agricultural chemicals on health and environment have generated a considerable interest in biological alternatives. This study aimed to test the insecticidal potential of fungus Aspergillus flavus and revealed its genotoxic and cytotoxic effects using Spodoptera litura (Fabricius) as a model. METHODS AND RESULTS: The fungus was isolated from the surface of the dead insect and investigated for its insecticidal potential against S. litura by bioassay studies. Significant increase in mortality, prolonged development period and reduced adult emergence in S. litura were observed in larva fed on diet supplemented with fungal extract. In addition, fungus was also found to cause oxidative stress, DNA damage and cell death. Significantly higher percentages of necrotic cells and DNA damage were observed in larvae treated with fungal extract. Furthermore, DNA repair studies predicted the longevity of toxic effects induced by fungus. Phytochemical and ultra-high performance liquid chromatography studies revealed the presence of phenolic compounds in the extract and liquid chromatography-mass spectrometry indicated it to be a non-aflatoxin strain of A. flavus. Fungal extract was less toxic to mammalian cell lines as compared to cytotoxic drug doxorubicin (DOX) in the MTT assay. CONCLUSION: The study highlights the insecticidal potential of A. flavus by revealing its genotoxicity and cytotoxicity causing potential. This is the first report showing the genotoxic and cytotoxic effects of the fungus A. flavus on S. litura. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides a useful insight to explore microbial agents as biopesticides in order to reduce various environmental as well as human health problems due to synthetic pesticides.

A retrospective review of 12 patients with trichotillomania treated in a psychodermatology service.

Trichotillomania is a condition characterized by the pulling of hair from anywhere on the body and is classified as an obsessive-compulsive and related disorder. Patients with hair disorders are commonly referred to psychodermatology services, and can represent a management challenge. Few publications exist that report outcomes for patients with trichotillomania in real clinical practice. We report 12 such patients seen within our own psychodermatology service, who were managed using a variety of treatment strategies. The rate of defaulting of appointments was high, but improvements were seen in patients engaging with services.

Effect of repeated intravenous lipopolysaccharide infusions on systemic inflammatory response and endometrium gene expression in Holstein heifers.

This study aimed to evaluate the effect of repeated intravenous lipopolysaccharide (LPS) infusions in nonlactating heifers on (1) the systemic proinflammatory state as measured by biomarkers in blood and plasma, and (2) endometrial gene expression of candidate transcripts on d 15 of gestation. Our hypothesis was that target transcripts related to a major functional group would be negatively modified in the preimplantation endometrium by the LPS treatments. In the first experiment (n = 13), a systemic proinflammatory state [defined as increased plasma concentrations of tumor necrosis factor (TNF)-α and haptoglobin for 2 wk] was established using 2 different sequential LPS infusion protocols. In the second experiment, heifers (n = 22; 11 mo of age) had their time of ovulation synchronized by a modified Ovsynch protocol and were enrolled in 1 of 2 treatments: control (CON; n = 11), which received sterile saline solution i.v., and LPS treatment (LPS; n = 11), submitted to repeated i.v. LPS injections (0.10, 0.25, 0.50, 0.75, 1.00, and 1.25 µg/kg) starting 2 d after artificial insemination (AI; d 0) and then every other day until d 15 after AI. At each LPS injection, rectal temperatures were measured hourly for 6 h. Blood samples were collected from d -1 to d 13 for analyses of progesterone, TNF-α, and haptoglobin in plasma, along with white blood cell (WBC) count and differential analysis. On d 15, endometrium tissue biopsies were taken and kept at -80°C until quantitative real-time PCR analysis of 30 target transcripts related to the immune system, adhesion molecules, and endometrium receptivity. Data were checked for normality and analyzed by repeated-measures ANOVA using PROC UNIVARIATE and PROC MIXED of SAS (SAS Institute Inc., Cary, NC). After each LPS injection, temperature was greater in the first 4 h in the LPS group compared with CON. Both TNF-α and haptoglobin increased in the LPS treatment with a significant treatment by day interaction. Total leukocyte count did not differ between treatments, but the differential count increased for neutrophils, band cells, and monocytes, and decreased for lymphocytes and eosinophils in LPS compared with CON. Progesterone concentrations in plasma did not differ between treatments during the experimental period. Out of 30 target genes analyzed, 3 transcripts were differentially expressed: indoleamine 2,3-dioxygenase (IDO; fold-change = 0.48) and pentraxin-3 (PTX3; fold-change = 0.38) were downregulated, whereas myxovirus-resistance protein (MX1; fold-change = 2.85) was upregulated in the LPS group. Sequential LPS injections were able to induce a prolonged systemic proinflammatory state, but effects on gene expression were limited to transcripts associated with the immune system. These results suggest that a mechanism for subfertility is linked to a proinflammatory state in dairy heifers.

Association of coronary artery calcium score with calcification and degree of stenosis: An autopsy study.

INTRODUCTION: Coronary artery disease (CAD) is a known cause of major cardiovascular events and calcium score (CS) has been developed as a marker of coronary atherosclerosis. Yet, the relationship between post mortem computed tomography (PMCT) CS with histologically observed calcification and the severity of coronary artery stenosis has not been widely explored and is still unclear. This study aims to determine the association between coronary artery PMCT CS with histologically observed calcification and degree of stenosis of coronary arteries in post-mortem cases. MATERIALS & METHODS: This was a cross-sectional study involving 101 subjects recruited from the National Institute of Forensic Medicine (IPFN) Hospital Kuala Lumpur (HKL) over a period of 15 months, from December 2012 until April 2014. PMCT CS of the coronary arteries was calculated using Agatston-Janowitz score. Histological presence of calcification was observed and the degree of stenosis was calculated using an image analysis technique. RESULTS: PMCT CS increased with increasing severity of stenosis (p<0.001). PMCT CS showed a positive correlation with the presence of calcification (r=-0.82, p<0.001). CONCLUSION: Calcium score is strongly associated with coronary artery calcification and the degree of luminal stenosis in post mortem subjects. Thus, PMCT may be useful as a non-invasive tool in diagnosing CAD in the event that an autopsy is not possible.

A clinicopathological study of upper aerodigestive tract cancers.

BACKGROUND: The upper aerodigestive tract (UAT) includes the nose and paranasal sinuses, oral cavity, pharynx, larynx, and salivary glands. Cancers of the UAT constitute approximately 4% of all malignancies. In this study, the varied nature of the UAT cancers was studied to find out their incidence, etiology, and clinicopathological correlations. MATERIALS AND METHODS: This prospective, observational, and clinicopathological study was conducted on 100 patients who were presented at outdoor in the Department of ENT, Government Medical College/Rajindra Hospital, Patiala, Punjab, India, from October 2016 to October 2018. Proven cases of UAT cancers were taken up and reviewed to gather data on multiple clinicopathological variables, such as age, sex, predisposing factors, and site of pathology. Histopathological differentiation was noted after conducting a biopsy. RESULTS: Most patients of UAT cancers were in the age group of 40-70 years. Maximum incidence was among males (82%) compared to females (28%). The most common predisposing factor was alcohol + smoking (28%), followed by alcohol + chewing tobacco (25%). The most common symptom in the oral cavity was ulcer and odynophagia (38%) each. In oropharyngeal cancers, dysphagia (92%) was the most common symptom. In laryngeal cancers, dyspnea (68%) and hoarseness of voice (32%) were the most common. The most common site involved in UAT cancers was the oral cavity (31%), followed by oropharynx (28%), larynx (22%), hypopharynx (7%), and salivary gland (5%). The most common histopathological type was squamous cell carcinoma (SCC) (90%). Most of the ulceroproliferative and exophytic growth was moderately differentiated SCC on histopathology. CONCLUSION: Studies are essential for education and awareness aimed at reducing exposure to habit-forming substances.

Prenatal profile of Pallister-Killian syndrome: Retrospective analysis of 114 pregnancies, literature review and approach to prenatal diagnosis.

Pallister-Killian syndrome (PKS) is a tissue limited mosaic disorder, characterized by variable degrees of neurodevelopmental delay and intellectual disability, typical craniofacial findings, skin pigmentation anomalies and multiple congenital malformations. The wide phenotypic spectrum of PKS in conjunction with the mosaic distribution of the i(12p) makes PKS an underdiagnosed disorder. Recognition of prenatal findings that should raise a suspicion of PKS is complicated by the fragmentation of data currently available in the literature and challenges in diagnosing a mosaic diagnosis on prenatal testing. Ultrasound anomalies, especially congenital diaphragmatic hernia, congenital heart defects, and rhizomelic limb shortening, have been related to PKS, but they are singularly not specific and are not present in all affected fetuses. We have combined prenatal data from 86 previously published reports and from our cohort of 114 PKS probands (retrospectively reviewed). Summarizing this data we have defined a prenatal growth profile and identified markers of perinatal outcome which collectively provide guidelines for early recognition of the distinctive prenatal profile and consideration of a diagnosis of PKS as well as for management and genetic counseling.

Intramammary infusion of lipopolysaccharide promotes inflammation and alters endometrial gene expression in lactating Holstein cows.

The objective of the current study was to evaluate the effects of 2 intramammary infusions of lipopolysaccharide (LPS) on inflammatory and reproductive parameters and endometrial gene expression of lactating Holstein cows. At 35 ± 7 d in milk, 20 cows were submitted to a Double Ovsynch program and randomly assigned to control (n = 11) and LPS (n = 9) treatments. Cows from the LPS treatment received 2 intramammary infusions of 25 µg of LPS after morning milking on d 5 and 10 post-AI, whereas control cows were infused with only saline. Blood samples were taken and ultrasound scanning of the ovaries was performed during the entire study before and after AI to determine haptoglobin, tumor necrosis factor-α, and progesterone concentrations as well as response to the hormonal protocol and corpus luteum diameter. Milk yield was evaluated and samples were taken for somatic cell count at 0, 10, 24, 34, and 96 h relative to each infusion. Rumen-reticular temperature was recorded using a rumen-reticular bolus logger and summarized hourly. On d 15 post-AI, uterine flushing for conceptus recovery and endometrial biopsies were performed. Samples of endometrium from cows with positive embryo recovery (control = 5; LPS = 6) were submitted to mRNA extraction and quantitative reverse-transcription PCR analysis of 96 target genes. Haptoglobin concentrations in plasma were greater for LPS treatment (control = 0.24 ± 0.07, LPS = 0.89 ± 0.06 optical density), but tumor necrosis factor-α concentrations were similar (control = 0.67 ± 0.11, LPS = 0.46 ± 0.11 ng/mL) between treatments. Lipopolysaccharide reduced milk yield after treatment (control = 34.3 ± 1.5, LPS = 29.4 ± 1.6 kg/d), whereas somatic cell count (log) was greater in LPS-treated cows until 34 h after infusions (control = 2.3 ± 0.1, LPS = 3.3 ± 0.1 cells/mL of milk). Rumen-reticular temperature of LPS cows was elevated between 5 and 10 h after each infusion compared with control cows (control = 39.5 ± 0.1, LPS = 40.1 ± 0.1°C). Progesterone concentration after AI was unaffected by treatment or pregnancy status as well as corpus luteum diameter and conceptus length on d 15. Lipopolysaccharide treatment altered the expression of 13 key genes in the endometrium (mostly upregulated), whereas another 17 tended to be modulated. Modified gene expression included genes related to immune response (PTX3 = 2.34-fold increase; IL6 = 3.42-fold increase; and TCN1 = 2.52-fold increase), adhesion molecules (CADM3 = 1.93-fold increase; MMP19 = 1.49-fold increase; EMMPRIN = 1.20-fold increase; SELL = 1.91-fold increase), Wnt signaling pathway (WNT2, FZD4, and FZD7, all <1.5-fold increase), and interferon-stimulated genes (BMP15 = 0.27-fold decrease; ISG15 = 2.17-fold increase, and MX2 = 2.23-fold increase). In summary, intramammary infusions of LPS were able to trigger an inflammatory response with no effect on corpus luteum diameter and concentration of progesterone in plasma. However, a limited but important set of modulations in the endometrium gene expression at d 15 of gestation was found.

CD10 expression pattern in prostatic adenocarcinoma: Elucidation of differences between Gleason's grades.

CD10, a transmembrane endopeptidase, has been shown to be lost as an early event in prostate cancer. We aimed at evaluating the pattern of expression of CD10 in various Gleason's grades of prostatic adenocarcinoma in comparison with nodular hyperplasia of prostate. This retrospective study included 30 cases of nodular hyperplasia and 30 of prostatic adenocarcinoma of various Gleason's grades. Immunohistochemical staining for CD10 was performed on all cases and positivity evaluated as percentage of cells as well as location (membranous or cytoplasmic or both). Of prostatic adenocarcinomas, grade 3 was seen in 10 foci, grade 4 in 28 and grade 5 in 22 foci. CD10 positivity in carcinoma was lower than in nodular hyperplasia, with the lowest positivity in grade 5. The pattern of expression of CD10 also changed from membranous in grade 3 to cytoplasmic in grade 5. Loss of CD10 expression appears to be associated with increasing tumour grade in carcinoma prostate and this can potentially be useful in stratification of such patients.