Cutaneous metastases as the presenting feature of lung cancer.

Lung cancer is the second most common cancer diagnosed in men and women worldwide with the highest mortality. A rare site of metastases for lung cancer is the skin. Typically, the diagnosis is secure prior to developing cutaneous metastases. We present a case of a man in his mid-70s who presented to dermatology with cutaneous metastases. We outline the presentation, diagnostic workup and management of this case. We also review the literature of cutaneous metastases in lung cancer; highlighting the clinical need for a timely accurate diagnosis and the implication in terms of prognosis.

Gait Characteristics and Cognitive Function in Middle-Aged Adults with and without Type 2 Diabetes Mellitus: Data from ENBIND.

Type 2 Diabetes Mellitus (T2DM) in midlife is associated with a greater risk of dementia in later life. Both gait speed and spatiotemporal gait characteristics have been associated with later cognitive decline in community-dwelling older adults. Thus, the assessment of gait characteristics in uncomplicated midlife T2DM may be important in selecting-out those with T2DM at greatest risk of later cognitive decline. We assessed the relationship between Inertial Motion Unit (IMUs)-derived gait characteristics and cognitive function assessed via Montreal Cognitive Assessment (MoCA)/detailed neuropsychological assessment battery (CANTAB) in middle-aged adults with and without uncomplicated T2DM using both multivariate linear regression and a neural network approach. Gait was assessed under (i) normal walking, (ii) fast (maximal) walking and (iii) cognitive dual-task walking (reciting alternate letters of the alphabet) conditions. Overall, 138 individuals were recruited (n = 94 with T2DM; 53% female, 52.8 ± 8.3 years; n = 44 healthy controls, 43% female, 51.9 ± 8.1 years). Midlife T2DM was associated with significantly slower gait velocity on both slow and fast walks (both p < 0.01) in addition to a longer stride time and greater gait complexity during normal walk (both p < 0.05). Findings persisted following covariate adjustment. In analyzing cognitive performance, the strongest association was observed between gait velocity and global cognitive function (MoCA). Significant associations were also observed between immediate/delayed memory performance and gait velocity. Analysis using a neural network approach did not outperform multivariate linear regression in predicting cognitive function (MoCA) from gait velocity. Our study demonstrates the impact of uncomplicated T2DM on gait speed and gait characteristics in midlife, in addition to the striking relationship between gait characteristics and global cognitive function/memory performance in midlife. Further studies are needed to evaluate the longitudinal relationship between midlife gait characteristics and later cognitive decline, which may aid in selecting-out those with T2DM at greatest-risk for preventative interventions.

DNA transfer to worn upper garments during different activities and contacts: An inter-laboratory study.

Cellular material derived from contact traces can be transferred via many direct and indirect routes, with the manner of contact and the time of transfer (in relation to the alleged crime-event) having an impact on whether DNA is recovered from the surface and a reportable profile generated. In an effort to acquire information on the transfer and recovery of DNA traces from clothing items worn during scenarios commonly encountered in casework, upper garments were worn during a normal working day before individuals were paired to embrace one another ('contact'), go on an outing together ('close proximity'), or individually asked to spend a day in another person's environment ('physical absence'). Each prescribed activity was repeated by sixteen individuals across four countries, and was the last activity performed before the garment was removed. Samples were collected from several areas of the upper garments and processed from DNA extraction through to profiling within the laboratory of the country in which the individual resided. Activities relating to the garment prior to and during wearing, including the prescribed activity, were recorded by the participant and considered during the interpretation of results. In addition to obtaining reference profiles from the wearer and their activity partner, DNA profiles from the wearers' close associates identified in the questionnaire were obtained to assess the impact of background DNA transferred prior to the prescribed activity. The wearer was typically, but not always, observed as the major contributor to the profiles obtained. DNA from the activity partner was observed on several areas of the garment following the embrace and after temporarily occupying another person's space. Particular areas of the garment were more prone to acquiring the hugging partner or office owner's DNA than others, and whether they were observed as the major or minor component was activity dependent. For each of the pairs, no DNA from the activity partner was acquired by the garments during the outing, even though both participants were in close proximity. This study provides empirical data on the transfer, persistence, prevalence and recovery of DNA from clothing items, and enables a better understanding of the mechanisms which lead to the transfer and detectability of DNA traces in different scenarios.

Prevalence of smell and taste dysfunction in a cohort of CoVID19 outpatients managed through remote consultation from a large urban teaching hospital in Dublin, Ireland.

BACKGROUND: Coronaviruses are known to precipitate disorders of smell and taste function. With the emerging global coronavirus disease (CoVID19) pandemic due to the severe acute respiratory syndrome coronavirus 2, early reports suggested that smell and taste dysfunction were clinical features of CoVID19. Our study aimed to investigate the prevalence of smell and taste disturbances in a cohort of CoVID19 positive patients who were isolated at home and being medically managed through telephone consultation. METHODS: This was a retrospective cross-sectional study conducted at St. James's Hospital Dublin, an urban 850 bed tertiary referral centre. 46 out of 50 CoVID19 positive patients, managed through a telephone clinic from the hospital infectious disease department, were assessed for olfactory and gustatory function loss. RESULTS: The median age of participants was 36.5 years (interquartile range (IQR) 27 - 48) and 19 (41%) were male. The majority (31; 67%) never smoked and 17 (37%) reported co-morbidities. Approximately half of patients reported some degree of smell (22; 48%) or taste (25; 54%) function loss. 13 patients (28%) reported a complete loss of sense of smell, 8 (17%) reported a complete loss of sense of taste and 7 (15%) reported simultaneous total loss of both. The median age of patients with any degree of smell disturbance was significantly lower than patients without (median 30.5 years (IQR 24 - 43.25) versus 41 years (IQR 28.25 - 52.75), p = 0.025). The median age of patients with any degree of taste disturbance was lower than those without, but did not reach statistical significance (median 34 years (IQR 24 - 45) versus 40 years (IQR 27.5 - 52.5) p = 0.174). CONCLUSION: Smell or taste dysfunction were present in our defined subgroup of patients. Further research is required in different population cohorts to build the evidence base for smell and taste dysfunction as clinical indicators of CoVID19 disease and to assess if these symptoms persist after disease resolution.

Peripheral Inflammation and Cognitive Performance in Middle-Aged Adults With and Without Type 2 Diabetes: Results From the ENBIND Study.

Midlife Type 2 Diabetes Mellitus (T2DM) is associated with a greater risk of dementia in later life. Peripheral inflammation and its impact on cognition is proposed as one of the pathological mechanisms mediating this link. However, studies have primarily focused on older individuals with established cognitive impairment and a long duration of T2DM. Importantly, knowledge of which individuals with midlife T2DM who are at greatest risk of later cognitive decline is lacking. We examined the cross-sectional relationship between serum levels of 8 pro-inflammatory markers (IL-1ß, IL-6, TNF-α, IL-8, MCP-1, CXCL10, IL-12p70, CRP) and performance on a detailed neuropsychological assessment battery in middle-aged adults with uncomplicated T2DM (N = 89; 52 ± 8.1 years, 47% female) and matched healthy controls (N = 50; 52 ± 8.3 years, 59% female). Linear regression was used to analyze associations between serum markers and cognitive performance in the overall cohort, followed by a T2DM∗protein concentration interaction analysis to identify any T2DM-specific effects. We observed a significant T2DM-specific association between serum TNF-α levels and scores on the Paired Associates Learning (PAL) task (ß: -3.16, SE: 1.32, p = 0.01, Std. Beta: -0.94), a task with significant working memory demands previously implicated in T2DM-related cognitive dysfunction. However, this did not persist on controlling for multiple testing. We provide exploratory evidence for a significant T2DM-specific relationship between serum TNF-α and memory performance. These findings require further replication and longitudinal analysis with the aim of selecting-out individuals with midlife T2DM at risk of future cognitive decline for potential preventative interventions.

Assessment of the transfer, persistence, prevalence and recovery of DNA traces from clothing: An inter-laboratory study on worn upper garments.

Among the various items recovered from crime scenes or persons involved in a crime event, clothing items are commonly encountered and submitted for forensic DNA sampling. Depending on the case circumstances and the activity-of-interest, sampling of the garment may concentrate on collecting DNA from the wearer, or from one or more offenders who have allegedly contacted the item and/or wearer. Relative to the targeted DNA, background DNA already residing on the item from previous contacts, or transferred during or after the crime event, may also be collected during sampling and observed in the resultant DNA profile. Given our limited understanding of how, and from where, background DNA is derived on clothing, research on the transfer, persistence, prevalence, and recovery (TPPR) of DNA traces from upper garments was conducted by four laboratories. Samples were collected from several areas of two garments, each worn on separate working or non-working days and individually owned by four individuals from each of the four laboratories, and processed from DNA extraction through to profiling. Questionnaires documented activities relating to the garment prior to and during wearing, and reference profiles were obtained from the wearer and their close associates identified in the questionnaire. Among the 448 profiles generated, variation in the DNA quantity, composition of the profiles, and inclusion/exclusion of the wearer and their close associates was observed among the collaborating laboratories, participants, garments worn on different occasions, and garment areas sampled.

Sharing data on DNA transfer, persistence, prevalence and recovery: Arguments for harmonization and standardization.

Sharing data between forensic scientists on DNA transfer, persistence, prevalence and recovery (TPPR) is crucial to advance the understanding of these issues in the criminal justice community. We present the results of a collaborative exercise on reporting forensic genetics findings given activity level propositions. This exercise outlined differences in the methodology that was applied by the participating laboratories, as well as limitations to the use of published data on DNA TPPR. We demonstrate how publication of experimental results in scientific journals can be further improved to allow for an adequate use of these data. Steps that can be taken to share and use these data for research and casework purposes are outlined, and the prospects for future sharing of data through publicly accessible databases are discussed. This paper also explores potential avenues to proceed with implementation and is intended to fuel the discussion on sharing data pertaining to DNA TPPR issues. It is further suggested that international standardization and harmonization on these topics will benefit the forensic DNA community as it has been achieved in the past with the harmonization of STR typing systems.

Wearer and non-wearer DNA on the collars and cuffs of upper garments of worn clothing.

During an investigation, the question of interest might be whether or not a person has worn a given garment. Wearer DNA studies have contributed to our knowledge and understanding of DNA transfer and persistence on items of worn clothing. However, there is currently no extensive dataset on DNA profiling outcomes from a variety of upper garments. Therefore we investigated the DNA profiling outcomes from the collars and/or cuffs of forty-four upper garments of worn clothing and determined the quantity of wearer and non-wearer DNA recovered. Interpretable DNA profiles were more likely to be obtained from collars than cuffs (84% versus 71%). The wearer was detected in all interpretable profiles and a major profile corresponding to the wearer was the most common outcome from both collar and cuff samples (48% and 50%, respectively). There was large variation in the amount of wearer DNA recovered and the average recovered was approximately 20 ng. Usually more wearer DNA was found on collars than cuffs of the same garment and, on average, more non-wearer DNA was found on cuffs than collars. No DNA was recovered from the cuffs of two garments despite these garments being worn for three and four hours, respectively. On one occasion a non-wearer contributed more DNA to a cuff sample than the wearer. We found no correlation between wearing time and the amount of wearer DNA recovered which indicates that other factors (e.g. shedder status and/or the manner of contact between the garment and skin) have a greater influence, than wearing time, on the amount of DNA transferred. However, there was a positive correlation between wearing time and the likelihood of obtaining interpretable profiles. This work has generated data to support the evaluation of DNA evidence from clothing and we provide a case example to demonstrate this.

An inter-laboratory comparison study on transfer, persistence and recovery of DNA from cable ties.

To address questions on the activity that led to the deposition of biological traces in a particular case, general information on the probabilities of transfer, persistence and recovery of cellular material in relevant scenarios is necessary. These figures may be derived from experimental data described in forensic literature when conditions relevant to the case were included. The experimental methodology regarding sampling, DNA extraction, DNA typing and profile interpretation that were used to generate these published data may differ from those applied in the case and thus the applicability of the literature data may be questioned. To assess the level of variability that different laboratories obtain when similar exhibits are analysed, we performed an inter-laboratory study between four partner laboratories. Five sets of 20 cable ties bound by different volunteers were distributed to the participating laboratories and sampled and processed according to the in-house protocols. Differences were found for the amount of retrieved DNA, as well as for the reportability and composition of the DNA profiles. These differences also resulted in different probabilities of transfer, persistence and recovery for each laboratory. Nevertheless, when applied to a case example, these differences resulted in similar assignments of weight of evidence given activity-level propositions.

Probability of detection of DNA deposited by habitual wearer and/or the second individual who touched the garment.

Available literature on the detection of transferred DNA does not address the interpretation issues in relation to who wore rather than touched the garment. To acquire a greater knowledge of the rate of detectable wearer, toucher and background DNA, 63 males wore their own underpants for 12h. The inside-waistband was handled by one of 11 female volunteers for 15s. The waist-band was mini-taped and subjected to DNA profiling with the AMPFℓSTR(®) NGM SElect™ kit. The findings show that on worn garments the probability of observing reportable DNA profiles is 61.9%. The wearer was detected as a single profile or part of a mixed profile in 50.8% of samples. When the wearer was present in a mixture, he was always observed as the major contributor. The toucher was detected on 11.1% of underpants. Reportable background DNA (non-wearer and non-toucher) occurs in 14.3% of samples and may affect the assessment of who wore the garment. Greater knowledge of the frequency of detection of reportable wearer DNA and/or toucher allows scientists to evaluate the likelihood of observing a matching profile if an individual wore a garment rather than touched it in disputed case scenarios.

Preeclampsia is associated with compromised maternal synthesis of long-chain polyunsaturated fatty acids, leading to offspring deficiency.

Obesity and excessive lipolysis are implicated in preeclampsia (PE). Intrauterine growth restriction is associated with low maternal body mass index and decreased lipolysis. Our aim was to assess how maternal and offspring fatty acid metabolism is altered in mothers in the third trimester of pregnancy with PE (n=62) or intrauterine growth restriction (n=23) compared with healthy pregnancies (n=164). Markers of lipid metabolism and erythrocyte fatty acid concentrations were measured. Maternal adipose tissue fatty acid composition and mRNA expression of adipose tissue fatty acid-metabolizing enzymes and placental fatty acid transporters were compared. Mothers with PE had higher plasma triglyceride (21%, P<0.001) and nonesterified fatty acid (50%, P<0.001) concentrations than controls. Concentrations of major n-6 and n-3 long-chain polyunsaturated fatty acids in erythrocytes were 23% to 60% lower (all P<0.005) in PE and intrauterine growth restriction mothers and offspring compared with controls. Subcutaneous adipose tissue Δ-5 and Δ-6 desaturase and very long-chain fatty acid elongase mRNA expression was lower in PE than controls (respectively, mean [SD] control 3.38 [2.96] versus PE 1.83 [1.91], P=0.030; 3.33 [2.25] versus 1.03 [0.96], P<0.001; 0.40 [0.81] versus 0.00 [0.00], P=0.038 expression relative to control gene [square root]). Low maternal and fetal long-chain polyunsaturated fatty acid concentrations in PE may be the result of decreased maternal synthesis.

The recovery and persistence of salivary DNA on human skin.

Salivary DNA is encountered in many crimes, such as sexual assaults and murders. In this study, saliva from three male donors was deposited on the skin of three female recipients. The amount of male salivary DNA remaining on the female skin was measured over a 96-h period using the Quantifiler™ Y Human Male DNA Quantification Kit. In eight of the nine experiments, a full male DNA profile matching the donor was obtained even after 96 h. In addition, the study showed that the concentration of salivary DNA varied from donor to donor and from day to day. The efficiency of two recovery methods, wet and dry swabbing and minitaping, was compared. The results indicate the tapelift method gave higher DNA recovery. This study also examined the secondary transfer of salivary DNA from skin to fabrics. Cotton and polyester give higher DNA transfer than leather.

A novel derivative for the assessment of urinary and salivary nitrate using gas chromatography/mass spectrometry.

Previous gas chromatography/mass spectrometry (GC/MS) methods for determining nitrate in biological samples involve either hazardous chemicals or produce multiple isomers that can be difficult to quantitate. Modification of these methods, by the nitration of mesitylene instead of benzene and in the presence of trifluoroacetic anhydride rather than sulphuric acid, should enable simple isotopic quantitation for use in tracer studies, for example, in the measurement of nitric oxide production. Desiccated urine and saliva samples, in addition to aqueous labelled and unlabelled nitrate standards, were treated with trifluoroacetic anhydride and mesitylene at 70 degrees C for 1 h, cooled, then sequentially washed with deionised water and aqueous sodium bicarbonate. The solution of nitromesitylene in mesitylene was separated, dried and analysed by GC/MS. The full mass spectra exhibited abundant ions at m/z 165 and 166 corresponding to the unlabelled and labelled molecular species of nitromesitylene, respectively. Selected ion monitoring of these masses for a series of gravimetrically prepared standards indicated good agreement with isotopic enrichments in the range 0.0625-5 mole % excess, and at nitrate concentrations within the physiological range of 0.078-2 mmol/L. Derivatised samples were stable with respect to isotopic enrichments and nitrate concentrations at -20 degrees C for up to 21 days and exhibited excellent repeatability. Nitration of mesitylene proved to be a simple and rapid method for the measurement of isotope ratios in aqueous nitrates by GC/MS, which has applications in tracer studies and in concentration determinations by isotope dilution techniques for nitric oxide production.

Biosensor developments: application to prostate-specific antigen detection.

Prostate-specific antigen (PSA) is the best serum marker currently available for the detection of prostate cancer and is the forensic marker of choice for determining the presence of azoospermic semen in some sexual assault cases. Most current assays for PSA detection are processed on large analyzers at dedicated testing sites, which require that samples be sent away for testing. This leads to delays in patient management and increased administration costs. The recent emphasis placed on the need for point-of-care patient management has led to the development of novel biosensor detection strategies that are suitable for the miniaturization of assays for various targets including PSA. This review highlights the current and novel analytical technologies used for PSA detection, which will benefit clinicians, patients and forensic workers in the future.

Gene expression profiling of serum- and interleukin-1 beta-stimulated primary human adult articular chondrocytes--a molecular analysis based on chondrocytes isolated from one donor.

In order to understand the cellular disease mechanisms of osteoarthritic cartilage degeneration it is of primary importance to understand both the anabolic and the catabolic processes going on in parallel in the diseased tissue. In this study, we have applied cDNA-array technology (Clontech) to study gene expression patterns of primary human normal adult articular chondrocytes isolated from one donor cultured under anabolic (serum) and catabolic (IL-1beta) conditions. Significant differences between the different in vitro cultures tested were detected. Overall, serum and IL-1beta significantly altered gene expression levels of 102 and 79 genes, respectively. IL-1beta stimulated the matrix metalloproteinases-1, -3, and -13 as well as members of its intracellular signaling cascade, whereas serum increased the expression of many cartilage matrix genes. Comparative gene expression analysis with previously published in vivo data (normal and osteoarthritic cartilage) showed significant differences of all in vitro stimulations compared to the changes detected in osteoarthritic cartilage in vivo. This investigation allowed us to characterize gene expression profiles of two classical anabolic and catabolic stimuli of human adult articular chondrocytes in vitro. No in vitro model appeared to be adequate to study overall gene expression alterations in osteoarthritic cartilage. Serum stimulated in vitro cultures largely reflected the results that were only consistent with the anabolic activation seen in osteoarthritic chondrocytes. In contrast, IL-1beta did not appear to be a good model for mimicking catabolic gene alterations in degenerating chondrocytes.

Hepatocyte growth factor/scatter factor is not a potent regulator of anabolic and catabolic gene expression in adult human articular chondrocytes.

Objective. Hepatocyte growth factor (HGF) has been reported to be present in articular cartilage and to be a potentially important inducing factor of anabolic and catabolic activity in chondrocytes. The aim of this study was to determine the expression levels of full length-functional-hgf and its receptor c-met in normal and osteoarthritic cartilage and the effect of HGF on anabolic and catabolic gene expression in adult human articular chondrocytes. Methods. Isolated adult human articular chondrocytes were stimulated for 48h with HGF (1, 10, and 100ng/ml). Synthesis of proteoglycans was determined by [(35)S]sulfate incorporation. mRNA levels for anabolic and catabolic genes as well as c-met and (functional) hgf were quantified using real-time PCR. Additionally, in situ mRNA expression levels of hgf and c-met were quantitatively measured from RNA directly isolated from normal and osteoarthritic adult human articular cartilage. Results. Proteoglycan synthesis in adult human articular chondrocytes was not stimulated by HGF nor was a selection of catabolic genes (collagenases and aggrecanases). Normal adult articular chondrocytes expressed only very low levels of hgf mRNA. Slightly higher levels of hgf were detected in chondrocytes isolated from osteoarthritic cartilage. Significant c-met expression was detected in both sample types. Conclusion. Despite the expression of its receptor c-met and its presence in articular cartilage, HGF does not appear to be a potent player in cartilage matrix turnover, at least not in terms of anabolic and catabolic gene expression in normal adult articular cartilage.