The adult shell matrix protein repertoire of the marine snail Crepidula is dominated by conserved genes that are also expressed in larvae.

Mollusca is a morphologically diverse phylum, exhibiting an immense variety of calcium carbonate structures. Proteomic studies of adult shells often report high levels of rapidly-evolving, 'novel' shell matrix proteins (SMPs), which are hypothesized to drive shell diversification. However, relatively little is known about the phylogenetic distribution of SMPs, or about the function of individual SMPs in shell construction. To understand how SMPs contribute to shell diversification a thorough characterization of SMPs is required. Here, we build tools and a foundational understanding of SMPs in the marine gastropod species Crepidula fornicata and Crepidula atrasolea because they are genetically-enabled mollusc model organisms. First, we established a staging system of shell development in C. atrasolea for the first time. Next, we leveraged previous findings in C. fornicata combined with phylogenomic analyses of 95 metazoan species to determine the evolutionary lineage of its adult SMP repertoire. We found that 55% of C. fornicata's SMPs belong to molluscan orthogroups, with 27% restricted to Gastropoda, and only 5% restricted at the species level. The low percentage of species-restricted SMPs underscores the importance of broad-taxon sampling and orthology inference approaches when determining homology of SMPs. From our transcriptome analysis, we found that the majority of C. fornicata SMPs that were found conserved in C. atrasolea were expressed in both larval and adult stages. We then selected a subset of SMPs of varying evolutionary ages for spatial-temporal analysis using in situ hybridization chain reaction (HCR) during larval shell development in C. atrasolea. Out of the 18 SMPs analyzed, 12 were detected in the larval shell field. These results suggest overlapping larval vs. adult SMP repertoires. Using multiplexed HCR, we observed five SMP expression patterns and three distinct cell populations within the shell field. These patterns support the idea that modular expression of SMPs could facilitate divergence of shell morphological characteristics. Collectively, these data establish an evolutionary and developmental framework in Crepidula that enables future comparisons of molluscan biomineralization to reveal mechanisms of shell diversification.

A chromosome-level genome for the nudibranch gastropod Berghia stephanieae helps parse clade-specific gene expression in novel and conserved phenotypes.

BACKGROUND: How novel phenotypes originate from conserved genes, processes, and tissues remains a major question in biology. Research that sets out to answer this question often focuses on the conserved genes and processes involved, an approach that explicitly excludes the impact of genetic elements that may be classified as clade-specific, even though many of these genes are known to be important for many novel, or clade-restricted, phenotypes. This is especially true for understudied phyla such as mollusks, where limited genomic and functional biology resources for members of this phylum have long hindered assessments of genetic homology and function. To address this gap, we constructed a chromosome-level genome for the gastropod Berghia stephanieae (Valdés, 2005) to investigate the expression of clade-specific genes across both novel and conserved tissue types in this species. RESULTS: The final assembled and filtered Berghia genome is comparable to other high-quality mollusk genomes in terms of size (1.05 Gb) and number of predicted genes (24,960 genes) and is highly contiguous. The proportion of upregulated, clade-specific genes varied across tissues, but with no clear trend between the proportion of clade-specific genes and the novelty of the tissue. However, more complex tissue like the brain had the highest total number of upregulated, clade-specific genes, though the ratio of upregulated clade-specific genes to the total number of upregulated genes was low. CONCLUSIONS: Our results, when combined with previous research on the impact of novel genes on phenotypic evolution, highlight the fact that the complexity of the novel tissue or behavior, the type of novelty, and the developmental timing of evolutionary modifications will all influence how novel and conserved genes interact to generate diversity.

Staged circumferential lumbar fusions have less intraoperative complications and shorter operative time with no difference in 30-, 90-, and 1-year complications: a propensity-matched cohort analysis of 190 patients.

BACKGROUND CONTEXT: Circumferential lumbar fusions (cLFs) are becoming more common with increasing and more minimally invasive anterior access techniques. Staging allows reassessment of indirect decompression and alignment prior to the posterior approach, and optimization of OR time management. Safety of staging has been well documented in deformity surgery but has yet to be delineated in less extensive, degenerative cLFs. PURPOSE: The purpose of this study is to compare perioperative complications and outcomes between staged versus single-anesthetic circumferential fusions in the lumbar spine. STUDY DESIGN: Propensity-matched comparative observational cohort. PATIENT SAMPLE: Patients who underwent cLFs for lumbar degenerative disease. OUTCOME MEASURES: In-hospital, 30-day, 90-day, and 1-year complications. METHODS: From 123 patients undergoing single-anesthetic and 154 patients undergoing staged cLF, 95 patients in each group were propensity-matched based on age, sex, BMI, ASA score, smoking, revision, and number of levels. We compared perioperative, 30-day, 90-day, and 1-year complications between the two cohorts. RESULTS: Mean days between stages was 1.58. Single-anesthetic cLF had longer total surgery time (304 vs 240 minutes, p<.001) but shorter total PACU total time (133 vs 196 minutes, p<.001). However, there was no difference in total anesthesia time (368 vs 374 minutes, p=.661) and total EBL (357 vs 320cc, p=.313). Intraoperative complications were nine incidental durotomies in the single-anesthetic and one iliac vein injury in the staged group (9% vs 1%, p=.018). There was no difference of in-hospital (38 vs 31, p=.291), 30-day (16 vs 23, p=.281), 90-day (10 vs 15, p=.391), 1-year complications (9 vs 12, p=.644), and overall cumulative 1-year complications (54 vs 56, p=.883) between the two cohorts. CONCLUSIONS: There is a decrease in total surgical time and intraoperative complications during staged compared with single-anesthetic cLF with no difference in in-hospital, 30-day, 90-day, and 1-year complications between approaches.

Normative Bone Mineral Density Measured on Computed Tomography Scans in Children and Adolescents.

STUDY DESIGN: Retrospective chart review. OBJECTIVE: To provide reference values for Hounsfield unit (HU) measured on computed tomography (CT) scans of children and adolescents. SUMMARY OF BACKGROUND DATA: Spine surgeons increasingly use HU on spine CT as a measure of bone mineral density (BMD). This has not been described in children and adolescents. PATIENTS AND METHODS: Pediatric patients who had a spine CT between 2012 and 2022 were identified. Patients who had more than 1 comorbidity or were syndromic were excluded. Using the bone window, 3 axial images (cephalad, middle, and caudal) of each vertebra were selected. In each axial view, the HU was measured using a "region of interest" (ROI) that included the total cancellous bone area and an ROI excluding the radiolucency present in the posterior vertebral body ("total" vs . "limited"). HU values were compared between total and limited areas and between the axial images at the cephalad, middle, and caudal levels. Each age category data were estimated and stratified. RESULTS: A total of 144 patients (79 females and 65 males) from 2 to 17 years old were included. Mean limited HU was consistently lower than total HU across all images and lumbar levels except for L1. Limited HU taken mid-vertebral body was also consistently lower than those taken cephalad or caudad. Mean limited HU across all ages including all levels was 227 ± 50 (range: 109-357). Stratifying by age showed a gradual decrease in BMD from age 2 to 10 followed by an increase. CONCLUSIONS: This is the first study to measure HU on lumbar CTs in children and adolescents. The technique of measuring BMD in adults should be modified in children by using an ROI that excludes the rarefaction present in the posterior vertebral body. Further studies are needed to evaluate the age-dependent changes in BMD seen in this study.

Opioid-sparing Anesthesia Decreases In-hospital and 1-year Postoperative Opioid Consumption Compared With Traditional Anesthesia: A Propensity-matched Cohort Study.

STUDY DESIGN: Propensity-matched cohort. OBJECTIVE: The aim of this study was to determine if opioid-sparing anesthesia (OSA) reduces in-hospital and 1-year postoperative opioid consumption. SUMMARY OF BACKGROUND DATA: The recent opioid crisis highlights the need to reduce opioid exposure. We developed an OSA protocol for lumbar spinal fusion surgery to mitigate opioid exposure. MATERIALS AND METHODS: Patients undergoing lumbar fusion for degenerative conditions over one to four levels were identified. Patients taking opioids preoperatively were excluded. OSA patients were propensity-matched to non-OSA patients based on age, sex, smoking status, body mass index, American Society of Anesthesiologists grade, and revision versus primary procedure. Standard demographic and surgical data, daily in-hospital opioid consumption, and opioid prescriptions 1 year after surgery were compared. RESULTS: Of 296 OSA patients meeting inclusion criteria, 172 were propensity-matched to non-OSA patients. Demographics were similar between cohorts (OSA: 77 males, mean age=57.69 yr; non-OSA: 67 males, mean age=58.94 yr). OSA patients had lower blood loss (326 mL vs. 399 mL, P =0.014), surgical time (201 vs. 233 min, P <0.001) emergence to extubation time (9.1 vs. 14.2 min, P< 0.001), and recovery room time (119 vs. 140 min, P =0.0.012) compared with non-OSA patients. Fewer OSA patients required nonhome discharge (18 vs. 41, P =0.001) compared with the non-OSA cohort, but no difference in length of stay (90.3 vs. 98.5 h, P =0.204). Daily opioid consumption was lower in the OSA versus the non-OSA cohort from postoperative day 2 (223 vs. 185 morphine milligram equivalents, P =0.017) and maintained each day with lower total consumption (293 vs. 225 morphine milligram equivalents, P =0.003) throughout postoperative day 4. The number of patients with active opioid prescriptions at 1, 3, 6, and 12 months postoperative was statistically fewer in the OSA compared with the non-OSA patients. CONCLUSIONS: OSA for lumbar spinal fusion surgery decreases in-hospital and 1-year postoperative opioid consumption. The minimal use of opioids may also lead to shorter emergence to extubation times, shorter recovery room stays, and fewer discharges to nonhome facilities.

Comparison of Bone Mineral Density in Children and Adolescents on CT Versus DEXA Scan.

STUDY DESIGN: Retrospective cross-sectional. OBJECTIVES: To determine associations between Hounsfield Units (HU) within a Region of interest (ROI) on CT scans and Dual Energy X-ray Absorptiometry (DEXA) measurements in children and adolescents. SUMMARY OF BACKGROUND DATA: HU on CT scans as a proxy for bone mineral density (BMD) is widely used in adults. However, the utility of CT as a proxy for BMD have not been evaluated in children and adolescents. METHODS: Patients younger than 18 years with a lumbar spine CT scan and a DEXA within 6 months of each other were identified. A ROI was used to measure the HU at each lumbar vertebral body on mid-axial cuts. Charts were reviewed for demographics, medical co-morbidities and DEXA reports. Correlation coefficients were calculated between mean DEXA Z-score and HU value. Patients were also stratified by Z-score (≥ -1.0, between -1.0 and -2.0, and ≤-2.0) and matched by age and BMI to a cohort of healthy children and compared. RESULTS: A moderate correlation between mean DEXA Z-score and mean HU on CT was found (r2=0.42, P<0.001). After matching for age and BMI, 21 patients in each group between the age of 4 and 17 years were analyzed. The mean HU value of the control group was 231.69. When stratified by Z-score (≥ -1.0, between -1.0 and -2.0, and ≤-2.0), the mean HU values were 244.59, 216.50, and 176.54, respectively. Patients with a Z-score of ≤-2.0 had a significantly lower mean HU than age matched controls. CONCLUSIONS: HU on lumbar CT in children and adolescents with DEXA Z-scores less than -2.0, were lower when compared to healthy age and BMI matched controls. This study suggests that HU on opportunistic CT scans of the spine may be used as a reasonable proxy for BMD in the pediatric population.

A chromosome-level genome for the nudibranch gastropod Berghia stephanieae helps parse clade-specific gene expression in novel and conserved phenotypes.

How novel phenotypes originate from conserved genes, processes, and tissues remains a major question in biology. Research that sets out to answer this question often focuses on the conserved genes and processes involved, an approach that explicitly excludes the impact of genetic elements that may be classified as clade-specific, even though many of these genes are known to be important for many novel, or clade-restricted, phenotypes. This is especially true for understudied phyla such as mollusks, where limited genomic and functional biology resources for members of this phylum has long hindered assessments of genetic homology and function. To address this gap, we constructed a chromosome-level genome for the gastropod Berghia stephanieae (Valdés, 2005) to investigate the expression of clade-specific genes across both novel and conserved tissue types in this species. The final assembled and filtered Berghia genome is comparable to other high quality mollusk genomes in terms of size (1.05 Gb) and number of predicted genes (24,960 genes), and is highly contiguous. The proportion of upregulated, clade-specific genes varied across tissues, but with no clear trend between the proportion of clade-specific genes and the novelty of the tissue. However, more complex tissue like the brain had the highest total number of upregulated, clade-specific genes, though the ratio of upregulated clade-specific genes to the total number of upregulated genes was low. Our results, when combined with previous research on the impact of novel genes on phenotypic evolution, highlight the fact that the complexity of the novel tissue or behavior, the type of novelty, and the developmental timing of evolutionary modifications will all influence how novel and conserved genes interact to generate diversity.

Risk Factors of Screw Malposition in Robot-Assisted Cortical Bone Trajectory: Analysis of 1344 Consecutive Screws in 256 Patients.

STUDY DESIGN: Retrospective single-center study using prospectively collected data. OBJECTIVE: To describe the incidence of and identify risk factors for intraoperative screw malposition secondary to skive or shift during robot-assisted cortical bone trajectory (RA-CBT) insertion. SUMMARY OF BACKGROUND DATA: RA-CBT screw malposition occurs through two distinct modes, skive or shift. Skive occurs when a downward force applied to the cannula, drill, tap, or screw, causes the instrument to deflect relative to its bony landmark. Shift is a change in position of the robot-assisted system relative to the patient after registration. METHODS: A consecutive series of patients older than 18 years who underwent RA-CBT screw placement between January 2019 and July 2022 were enrolled. Baseline demographic and surgical data, Hounsfield Units (HU) at L1, and vertebral shape related to screw planning were collected. Skive or shift was recorded in the operating room on a data collection form. RESULTS: Of 1344 CBT screws in 256 patients, malposition was recognized intraoperatively in 33 screws (2.4%) in 27 patients (10.5%); 19 via skive in 17 and 14 via shift in 10 patients. These patients had higher BMI than patients without malposition (33.0 kg/m2 vs 30.5 kg/m2, P=0.037). Patients with skive had higher HU (178.2 vs 145.2, P=0.035), compared to patients with shift (139.2 vs 145.2, P=0.935) and patients without screw malposition. More than half of screw malposition was observed at the UIV. At the UIV, if the screw's overlap to the bone surface at the insertion point was decreased, skive was more likely (57% vs 87%, P<0.001). No patients were returned to the operating room for screw revision. CONCLUSIONS: Intraoperative screw malposition occurred in 2.4% of RA-CBT. High BMI was associated with screw malposition, regardless of etiology. Skive was associated with high HU and decreased screw overlap to bone surface at the insertion point.

Quantitative Romberg on a Force Plate: Objective Assessment Before and After Surgery for Cervical Spondylotic Myelopathy.

STUDY DESIGN: Longitudinal Observational Cohort. OBJECTIVES: The purpose of this study was to evaluate the utility of Quantitative Romberg measurements as pre- and post-op balance outcome measures. SUMMARY OF BACKGROUND DATA: Cervical Spondylotic Myelopathy (CSM) is characterized by balance deficiencies produced by impaired proprioception. Evaluation is subjective and binary physical exam findings lack the precision to assess postoperative outcome improvement. METHODS: CSM patients were prospectively enrolled to undergo pre- and postoperative Quantitative Romberg tests on a force plate to record center of pressure (COP) motion for 30 seconds with eyes open followed by eyes closed. Revision cases were excluded. Kinematics of COP movement parameters were compared between pre- and postoperative state for each patient. RESULTS: Twenty-seven CSM patients were enrolled and completed both pre/post testing. Mean age was 60.0 years with 13 (48%) males, 9 (33%) smokers. Mean number of surgical levels was 2.48. The minimum mean follow-up was six months. There was a statistically significant improvement in eyes closed after surgery compared to pre-operative for total COP motion (523.44 cm vs. 387.00 cm, P<0.001), average sway speed (17.41 cm/s vs. 13.00 cm/s, P<0.001) and total lateral COP motion (253.44 cm vs. 186.70 cm, P<0.001). There was no statistically significant improvement in mJOA (13.29 vs. 14.29, P=0.28). CONCLUSION: CSM balance findings on Quantitative Romberg testing significantly improves postoperatively in patients with CSM. These findings support this testing as representative of proprioceptive balance deficiencies seen in CSM. Quantitative Romberg may be used as an objective measure of clinical outcome and assist in stratification of surgical interventions, surgery timing and technique.

Comparison of revision surgery for pseudarthrosis with or without adjacent segment disease after anterior cervical discectomy and fusion.

Background: Patients with a pseudarthrosis after anterior cervical discectomy and fusion (ACDF) may have concurrent adjacent segment disease (ASD). Although prior studies have shown posterior cervical decompression and fusion (PCDF) is effective in repairing pseudarthrosis, improvement in patient reported outcomes (PROs) has been marginal. The aim of this study is to evaluate the effectiveness of PCDF in achieving symptom relief in patients with pseudarthrosis after ACDF and whether that is altered by the additional treatment of ASD. Methods: Thirty-two patients with pseudarthrosis were compared with 31 patients with pseudarthrosis and concurrent ASD after ACDF who underwent revision PCDF with a minimum 1-year follow-up. Primary outcomes measures included the neck disability index (NDI), and numerical rating scale (NRS) scores for neck and arm pain. Secondary measures included estimated blood loss (EBL), operating room (OR) time, and length of stay. Results: Demographics between cohorts were similar, however there was a significantly higher mean body mass index (BMI) in the group with concurrent ASD (32.23 vs. 27.76, p=.007). Patients with concurrent ASD had more levels fused during PCDF (3.7 vs. 1.9, p<.001), greater EBL (165 cc vs. 106 cc, p=.054), and longer OR time (256 minutes vs. 202 minutes, p<.000). Preoperative PROs for NDI (56.7 vs. 56.5, p=.954), NRS arm pain (5.9 vs. 5.7, p=.758), and NRS neck pain (6.6 vs. 6.8, p=.726) were similar in both cohorts. At 12 months patients with concurrent ASD experienced a slightly greater, but not statistically significant, improvement in PROs (Δ NDI 4.40 vs. -1.44, Δ NRS neck pain 1.17 vs. 0.42, Δ NRS arm pain 1.28 vs. 0.10, p=.107). Conclusions: PCDF is a standard procedure for treatment of pseudarthrosis following ACDF, however improvements in PROs are marginal. Slightly greater improvements were seen in patients whose indication for surgery also included concurrent ASD, rather than a diagnosis of pseudarthrosis alone.

Spatial-temporal expression analysis of lineage-restricted shell matrix proteins reveals shell field regionalization and distinct cell populations in the slipper snail Crepidula atrasolea.

Molluscs are one of the most morphologically diverse clades of metazoans, exhibiting an immense diversification of calcium carbonate structures, such as the shell. Biomineralization of the calcified shell is dependent on shell matrix proteins (SMPs). While SMP diversity is hypothesized to drive molluscan shell diversity, we are just starting to unravel SMP evolutionary history and biology. Here we leveraged two complementary model mollusc systems, Crepidula fornicata and Crepidula atrasolea , to determine the lineage-specificity of 185 Crepidula SMPs. We found that 95% of the adult C. fornicata shell proteome belongs to conserved metazoan and molluscan orthogroups, with molluscan-restricted orthogroups containing half of all SMPs in the shell proteome. The low number of C. fornicata -restricted SMPs contradicts the generally-held notion that an animal’s biomineralization toolkit is dominated by mostly novel genes. Next, we selected a subset of lineage-restricted SMPs for spatial-temporal analysis using in situ hybridization chain reaction (HCR) during larval stages in C. atrasolea . We found that 12 out of 18 SMPs analyzed are expressed in the shell field. Notably, these genes are present in 5 expression patterns, which define at least three distinct cell populations within the shell field. These results represent the most comprehensive analysis of gastropod SMP evolutionary age and shell field expression patterns to date. Collectively, these data lay the foundation for future work to interrogate the molecular mechanisms and cell fate decisions underlying molluscan mantle specification and diversification.

Paediatric Day Case Tonsillectomy - Audit of a New Programme.

Aims The primary objective of this audit was to assess 30-day unplanned admission or readmission rates following day case tonsillectomy. Secondary objectives included assessing demographic characteristics of patients and surgical and anaesthetic techniques employed. Methods Retrospective chart review was performed. Results 34 tonsillectomies or adenotonsillectomies were performed for children aged between 5 and 17 years. A total of six patients (17.6%) were either admitted from the day ward or readmitted within 30 days. This was not statistically significantly greater than the maximum acceptable rate of 15% as recommended by ENT UK and the Royal College of Surgeons (95% confidence intervals 4.8% - 30.5%, p = 0.33). Of these admissions, one (2.9%) was due to nausea and inadequate oral intake, one (2.9%) was due to clinical concerns regarding sleep apnoea and four readmissions (11.8%) were due to bleeding. Anaesthesia techniques used varied considerably. Conclusion Further multidisciplinary co-operation and standardisation of care may help to improve this programme and reduce unplanned admission and readmission rates.

Attitudes and Knowledge of Healthcare Professionals Regarding Organ Donation. A Survey of the Saolta University Health Care Group.

Organ donation saves lives and healthcare professionals (HCPs) play a vital role in that process. Therefore, the purpose of this study was to assess the attitudes and level of knowledge of HCPs regarding organ donation. An online anonymous self-administered questionnaire containing 40 questions on organ donation using google forms was created. The survey was distributed to HCPs working in the Saolta University Health Care Group. A hundred and thirty-nine responses were received giving a response rate of 11.8%. HCPs willingness to donate their organs was at 93% compared to 97% willing to receive a transplant. More HCPs understood or had knowledge of the term donation after brain death (64%) than donation after circulatory death (49%). HCPs working in intensive care knew more about the management of brain dead donors than other specialties (p<0.0001). Over 60% of HCPs when asked either disagreed or strongly disagreed with the adequacy of training in organ donation and transplant. Overall, HCPs surveyed had positive attitudes towards organ donation but there was a lack of knowledge particularly among non-intensive care professionals. This study highlights the need to increase awareness along with implementation of educational programmes among HCPs regarding organ donation and transplant.

Liaison psychiatry professionals' views of general hospital care for patients with mental illness: The care of patients with mental illness in the general hospital setting.

OBJECTIVE: Explore the experiences of liaison psychiatry professionals, to gain a greater understanding of the quality of care patients with mental illness receive in the general hospital setting; the factors that affect the quality of care; and their insights on interventions that could improve care. METHODS: A survey questionnaire and qualitative in depth interviews were used to collect data. Data collection took place at the Royal College of Psychiatrists Faculty of Liaison Psychiatry Annual conference. Qualitative analysis was done using thematic analysis. RESULTS: Areas of concern in the quality of care of patients with co-morbid mental illness included 'diagnostic overshadowing', 'poor communication with patient', 'patient dignity not respected' and 'delay in investigation or treatment'. Eleven contributing factors were identified, the two most frequently mentioned were 'stigmatising attitudes of staff towards patients with co-morbid mental illness' and 'complex diagnosis'. The general overview of care was positive with areas for improvement highlighted. Interventions suggested included 'formal education' and 'changing the liaison psychiatry team'. CONCLUSION: The cases discussed highlighted several areas where the quality of care received by patients with co-morbid mental illness is lacking, the consequences of which could be contributing to physical health disparities. It was acknowledged that it is the dual responsibility of both the general hospital staff and liaison staff in improving care.

Micropore closure kinetics are delayed following microneedle insertion in elderly subjects.

Transdermal delivery is an advantageous method of drug administration, particularly for an elderly population. Microneedles (MNs) allow transdermal delivery of otherwise skin-impermeable drugs by creating transient micropores that bypass the barrier function of the skin. The response of aging skin to MNs has not been explored, and we report for the first time that micropore closure is delayed in elderly subjects in a manner that is dependent upon MN length, number, and occlusion of the micropores. Twelve control subjects (25.6±2.8years) and 16 elderly subjects (77.3±6.8years) completed the study. Subjects were treated with MNs of 500µm or 750µm length, in arrays containing 10 or 50 MNs. Impedance measurements made at baseline, post-MN insertion, and at predetermined time points demonstrated that restoration of the skin barrier is significantly slower in elderly subjects under both occluded and non-occluded conditions. This was confirmed via calculation of the total permeable area created by the micropores (which would approximate the area available for drug delivery), as well as calculation of the micropore half-life. This pilot study demonstrates that longer timeframes are required to restore the barrier function of aged skin following MN insertion, suggesting that drug delivery windows could be longer following one treatment with a MN array.

Age-dependent variation in cytokines, chemokines, and biologic analytes rinsed from the surface of healthy human skin.

In the skin, aging is associated with overall epidermal thinning, decreased barrier function, and gradual deterioration of the epidermal immune response. However, the presence and role of cytokines, chemokines, and biologic analytes (CCBAs) in immunosenescence are not known. Here we identified age-related changes in skin properties and CCBAs from stratum corneum of healthy human subjects, providing a means to utilize CCBAs as benchmarks for aging skin health. Transepidermal water loss and a(*) (skin redness) decreased in an age-dependent manner, and were significantly lower (p < 0.05) in Groups 2 (56.6 ± 4.6 years) and 3 (72.9 ± 3.0 years) vs. Group 1 (24.3 ± 2.8 years). In skin wash fluid, 48 CCBAs were detected; seven were significantly lower (p < 0.05) in Groups 2 and 3: EGF, FGF-2, IFNα2, IL-1RA, HSA, keratin-6, and involucrin; cortisol was significantly higher (p < 0.05) in Groups 2 and 3. Our results correspond with the pro-inflammatory shift that occurs with immunosenescence and also provides basis for understanding the inflammatory changes in normal aging skin.

In vitro bioactive properties of intact and enzymatically hydrolysed whey protein: targeting the enteroinsular axis.

Enzymatically hydrolysed milk proteins have a variety of biofunctional effects some of which may be beneficial in the management of type 2 diabetes mellitus. The purpose of this study was to evaluate the effect of commercially available intact and hydrolysed whey protein ingredients (DH 32, DH 45) on markers of the enteroinsular axis (glucagon like peptide-1 secretion, dipeptidyl peptidase IV inhibition, insulin secretion and antioxidant activity) before and after simulated gastrointestinal digestion (SGID). A whey protein hydrolysate, DH32, significantly enhanced (P < 0.05) insulin secretion from BRIN BD11 ß-cells compared to the positive control (16.7 mM glucose and 10 mM Ala). The whey protein hydrolysates inhibited dipeptidyl peptidase IV activity, yielding half maximal inhibitory concentration values (IC50) of 1.5 ± 0.1 and 1.1 ± 0.1 mg mL(-1) for the DH 32 and DH 45, samples respectively, and were significantly more potent than the intact whey (P < 0.05). Enzymatic hydrolysis of whey protein significantly enhanced (P < 0.05) its antioxidant activity compared to intact whey, as measured by the oxygen radical absorbance capacity assay (ORAC). This antioxidant activity was maintained (DH 32, P > 0.05) or enhanced (DH 45, P < 0.05) following SGID. Intact whey stimulated GLP-1 secretion from enteroendocrine cells compared to vehicle control (P < 0.05). This data confirm that whey proteins and peptides can act through multiple targets within the enteroinsular axis and as such may have glucoregulatory potential.

Optimization of impedance spectroscopy techniques for measuring cutaneous micropore formation after microneedle treatment in an elderly population.

PURPOSE: The objective of this study was to optimize a reproducible impedance spectroscopy method in elderly subjects as a means to evaluate the effects of microneedles on aging skin. METHODS: Human volunteers were treated with microneedles at six sites on the upper arm. Repeated impedance measurements were taken pre- and post-microneedle insertion. Two electrode types were evaluated (dry vs. gel), using either light or direct pressure to maintain contact between the electrode and skin surface. Transepidermal water loss (TEWL) was measured as a complementary technique. RESULTS: Five control subjects and nine elderly subjects completed the study. Microneedle insertion produced a significant decrease in impedance from baseline in all subjects (p < 0.05, regardless of electrode type or pressure application), confirming micropore formation. This was supported by a complementary significant increase in TEWL (p < 0.05). The gel*direct condition produced the lowest variability between measurements, as demonstrated by a coefficient of variation of 3.8% and 3.5% (control and elderly subjects, respectively). This was lower than variation between TEWL measurements at the same sites: 19.8% and 21.6% (control and elderly subjects, respectively). CONCLUSIONS: Impedance spectroscopy reproducibly measures micropore formation in elderly subjects, which will be essential for future studies describing microneedle-assisted transdermal delivery in aging populations.

Robotic gastric electrical stimulator placement: technique and literature review.

Gastroparesis in a chronic setting is a disorder that results in diminished quality of life. Laparoscopic gastric electrical stimulator (GES) placement is now being performed in patients with medically refractory gastroparesis. During this procedure, a significant amount of suturing is required to anchor the electrodes to the gastric wall. Robotic surgery may provide surgeons with several technical and ergonomic advantages during this procedure, when compared with a standard laparoscopic approach. The aim of this study is to present a case and review the technique and literature for robotic placement of GES. This report demonstrates the safety and feasibility of robotic GES placement.

Antimicrobial Activity of Chemokine CXCL10 for Dermal and Oral Microorganisms.

CXCL10 (IP-10) is a small 10 kDa chemokine with antimicrobial activity. It is induced by IFN-γ, chemoattracts mononuclear cells, and promotes adhesion of T cells. Recently, we detected CXCL10 on the surface of the skin and in the oral cavity. In the current study, we used broth microdilution and radial diffusion assays to show that CXCL10 inhibits the growth of Escherichia coli, Staphylococcus aureus, Corynebacterium jeikeium, Corynebacterium striatum, and Candida albicans HMV4C, but not Corynebacterium bovis, Streptococcus mutans, Streptococcus mitis, Streptococcus sanguinis, Fusobacterium nucleatum, Aggregatibacter actinomycetemcomitans, Poryphromonas gingivalis, or C. albicans ATCC 64124. The reason for the selective antimicrobial activity is not yet known. However, antimicrobial activity of CXCL10 may be related to its composition and structure, as a cationic 98 amino acid residue molecule with 10 lysine residues, 7 arginine residues, a total net charge of +11, and a theoretical pI of 9.93. Modeling studies revealed that CXCL10 contains an α-helix at the N-terminal, three anti-parallel ß-strands in the middle, and an α-helix at the C-terminal. Thus, CXCL10, when produced on the surface of the skin or in the oral cavity, likely has antimicrobial activity and may enhance innate antimicrobial and cellular responses to the presence of select commensal or opportunistic microorganisms.