"Just add small molecules" cell-free protein synthesis: Combining DNA template and cell extract preparation into a single fermentation.

Cell-free protein synthesis (CFPS) is a versatile biotechnology platform enabling a broad range of applications including clinical diagnostics, large-scale production of officinal therapeutics, small-scale on-demand production of personal magistral therapeutics, and exploratory research. The shelf stability and scalability of CFPS systems also have the potential to overcome cost and infrastructure challenges for distributing and using essential medical tests at home in both high- and low-income countries. However, CFPS systems are often more time-consuming and expensive to prepare than traditional in vivo systems, limiting their broader use. Much work has been done to lower CFPS costs by optimizing cell extract preparation, small molecule reagent recipes, and DNA template preparation. In order to further reduce reagent cost and preparation time, this work presents a CFPS system that does not require separately purified DNA template. Instead, a DNA plasmid encoding the recombinant protein is transformed into the cells used to make the extract, and the extract preparation process is modified to allow enough DNA to withstand homogenization-induced shearing. The finished extract contains sufficient levels of intact DNA plasmid for the CFPS system to operate. For a 10 mL scale CFPS system expressing recombinant sfGFP protein for a biosensor, this new system reduces reagent cost by more than half. This system is applied to a proof-of-concept glutamine sensor compatible with smartphone quantification to demonstrate its viability for further cost reduction and use in low-resource settings.

Content Evaluation of Residency Websites for All 159 Anesthesiology ACGME Programs in the USA.

Background: The shift to virtual interviews during the COVID-19 pandemic has elevated the vital role of Accreditation Council for Graduate Medical Education residency program websites in conveying information to applicants. The purpose of our study was to assess the recruitment, education, and diversity and inclusion content on websites for anesthesiology residency programs. Second, we aimed to test the hypothesis that the content scores of websites are higher in programs with more National Institutes of Health funding, in programs that are university-based versus community-based, and in larger programs, as measured by number of residents. Methods: Two independent reviewers evaluated the websites of the 159 anesthesiology residency programs accredited by the Accreditation Council for Graduate Medical Education for the presence (yes/no) of 12 recruitment, 6 education, and 8 diversity and inclusion criteria. Multiple linear regression was used to determine which program factors were most associated with total website content score. Results: Anesthesiology residency program websites contained a mean of 12.9 (SD = 3.4; range, 3-21) of the 26 study-defined criteria. The most common recruitment, education, and diversity and inclusion criteria were, respectively, program description, rotation information, and community demographics. Controlling for program factors, a university-based affiliation (P = .016) was associated with higher website content scores. Conclusions: There is large variation in the recruitment, education, and diversity and inclusion content on anesthesiology residency program websites nationally. Since program websites averaged only half of criteria, this may provide an impetus for programs to modify their websites, which may inform applicant decisions about which programs align with their training and career goals.

Using Pitch-Tracking Data to Identify Risk Factors for Medial Ulnar Collateral Ligament Reconstruction in Major League Baseball Pitchers.

Background: Approximately 25% of Major League Baseball (MLB) pitchers undergo medial ulnar collateral ligament reconstruction (UCLR) during their careers. Purpose: To identify risk factors for UCLR that are specific to 2 subgroups of MLB pitchers: right (RHPs)- versus left-handed pitchers (LHPs) and starting (SPs) versus relief pitchers (RPs). Study Design: Case-control study; Level of evidence, 3. Methods: We included 109 MLB pitchers who had undergone UCLR between 2007 and 2019 and had sufficient preinjury data in the 3 years before surgery (T3, T2, T1). A 2:1 matched control cohort was selected for comparison. Pitch velocity, release location, and ball movement were compared between the UCLR and control cohorts in both subgroups in the years before surgery (RHPs vs LHPs and SPs vs RPs). Binary logistic regression was used to identify independent risk factors for UCLR. Results: The mean horizontal release location for the UCLR group was 5.8 cm more lateral than for the control group (P = .028). For all pitchers, every 2.5-cm lateral shift in release location in the years leading up to UCLR equated to a 3.7% increase in the odds of UCLR. For RPs, this risk was more substantial: a 5.8% increase in odds per 2.5 cm. SPs in the UCLR group demonstrated significantly different T1 horizontal release locations compared with SPs in the control group, though not to a statistically significant change over the 3 years before surgery. However, in the 3 years before surgery, the horizontal release location for RPs in the UCLR group moved 2.1 cm more lateral, as compared with 2.7 cm more medial for RPs in the control group (P = .007). For LHPs, a decrease in mean pitch velocity by 1 mph (1.6 km/h) in the years leading up to surgery increased the odds of UCLR by 45%. Conclusion: Increasing lateralization of release point in the years before surgery increased the risk of UCLR, specifically for relievers. Our findings add to the growing body of evidence that release location is an important variable in analyzing the risk of UCLR in MLB and that risk stratification may be dependent on pitcher characteristics such as position, handedness, and weight.

An Analysis of the Diversity and Inclusion Content Featured on Ophthalmology Residency Program Web Sites.

Introduction Ophthalmology departments have been stated to be among the least diverse clinical departments at United States medical schools. Improvement requires recruiting a pipeline of diverse trainees. Residency program Web sites represent a potential diversity and inclusion recruitment tool. This study assesses how ophthalmology residency program Web sites demonstrate a commitment to diversity and inclusion. Methods We analyzed the diversity and inclusion content of 116 ophthalmology residency program Web sites in April 2021. Main outcome measures were the presence of 12 diversity and inclusion elements on program Web sites, based on prior work and Accreditation Council for Graduate Medical Education guidelines: nondiscrimination statement, diversity and inclusion message, community resources, extended faculty or resident biographies (including hobbies, etc.), faculty photos, resident photos, additional financial resources for trainees, wellness resources, mental health resources, health disparities/community engagement, and diversity council. We used Mann-Whitney U and Kruskal-Wallis tests to assess whether residency program characteristics such as rank, size, university affiliation, and geographic location were associated with commitment to diversity and inclusion on affiliated residency Web sites. Results Ophthalmology residency program Web sites included a mean of 4.4 ± 2.1 diversity elements. Sixteen percent of programs featured more than half (7 + ) of the evaluated diversity elements. The most featured common diversity elements included resident photos (85.3%), faculty photos (78.4%), and community resources (64.3%). Extended faculty biographies (2.6%), mental health resources (9.5%), and diversity council information (11.2%) were less commonly showcased. Top-ranked programs (7.6 ± 1.8, p < 0.0001) and university-based/-affiliated programs (4.7 ± 2.8, p = 0.0039) displayed more diversity elements than lower-ranked (4.1 ± 1.8) and community-based programs (2.8 ± 1.7). Conclusion Most ophthalmology residency program Web sites feature less than half of the 12 diversity and inclusion elements included in this study, suggesting room for improvement. By drawing attention to program diversity and inclusion efforts, Web sites offer a potential tool for residency programs to consider in their recruitment efforts for diverse trainees. Incorporating the diversity and inclusion elements analyzed in this study represents a low-burden way to signal a greater commitment to diversity that could help programs recruit diverse applicants.

Online public interest in common malignancies and cancer screening during the COVID-19 pandemic in the United States.

BACKGROUND AND AIM: The COVID-19 pandemic was declared a national emergency in the United States in March 2020. The Centers for Medicare and Medicaid Services subsequently released recommendations that health-care facilities temporarily delay elective surgeries and non-essential medical procedures. Disruptions to medical care significantly impacted cancer patients, with cancer screenings halted and nonurgent cancer surgeries postponed as health-care facilities shifted resources toward the COVID-19 pandemic. Although it has been reported that cancer screening rates decreased dramatically in the United States in 2020, it is unclear whether this trend was driven by factors related to public interest in cancer and/or cancer screening as opposed to other factors such as clinical backlogs, pandemic-related policies, and/or resource limitations. The purpose of this study was to use the Google Trends tool to evaluate public interest in six common malignancies and four common cancer screening methods during the COVID-19 pandemic. METHODS: We used the Google Trends tool to quantify public interest in six different malignancies (Breast Cancer, Colon Cancer, Lung Cancer, Prostate Cancer, Thyroid Cancer, and Cervical Cancer) and four cancer screening methods (Pap Smear, Lung Cancer Screening, Mammogram, and Colonoscopy) in the United States during the COVID-19 pandemic. Welch's t-tests were used to compare monthly search volumes during the COVID-19 pandemic (2020) to the 4 years before the pandemic (2016 - 2019) for all ten search terms included in our study. We used Benjamini-Hochberg to adjust raw p values to account for multiple statistical comparisons. The level of statistical significance was defined by choosing a false discovery rate of 0.05. RESULTS: Our results indicate significantly reduced interest in all malignancies studied at the beginning of the COVID-19 pandemic. Public interest in ['Breast Cancer'], ['Colon Cancer'], ['Lung Cancer'], ['Thyroid Cancer'], and ['Cervical Cancer'] significantly decreased in the months of March, April, May, and June 2020 when compared with public interest in 2016-2019. Public interest in cancer screening methods such as ['Pap Smear'], ['Lung Cancer Screening'], ['Mammogram'], and ['Colonoscopy'] significantly deceased in the months of April and May compared to 2016 - 2019 values. However, decreased public interest in cancer screening methods was temporary, with Google search volumes returning to pre-pandemic levels in June 2020 - December 2020. CONCLUSION: There was significantly reduced public interest in both common malignancies and cancer screening methods at the beginning of the COVID-19 pandemic in the United States. However, after an initial decline, public interest as indicated by Google search volumes quickly returned to pre-pandemic levels in the second half of the calendar year 2020. In addition, trends in public interest in cancer screening as indicated by Google search volumes aligned with cancer screening uptake rates in the United States during the study period. This finding suggests that Google Trends may serve as an effective tool in gauging the public's interest in cancer and/or cancer screenings in the United States, which makes it a valuable resource that can be used to inform decisions aimed at improving cancer screening rates in the future. RELEVANCE FOR PATIENTS: The Google Trends tool can be used to measure public interest in various malignancies and their associated screening methods. Google Trends data may be used to inform measures aimed at improving cancer screening uptake.

Differentiating Upper Extremity Necrotizing Soft Tissue Infection From Serious Cellulitis and Abscess.

Introduction Necrotizing soft tissue infection (NSTI) of the upper extremity (UE) is a rapidly progressing infection that requires early diagnosis and emergent treatment to decrease risks of loss of limb or life. Clinical presentation, particularly of early NSTI, can appear similar to serious cellulitis or abscess. The purpose of this study was to identify factors that are associated with NSTI rather than serious cellulitis and abscess to differentiate patients with similar clinical presentations. Methods This study uses a retrospective cohort design that compares patients ultimately diagnosed with UE NSTI versus those diagnosed with UE serious cellulitis or abscess. Cohorts were matched using the Laboratory Risk Indicators for Necrotizing Fasciitis (LRINEC) score in the setting of UE soft tissue infection. Laboratory values, vital signs, subjective symptoms, and social factors including substance abuse and domiciled status were recorded. Continuous variables were compared using the Mann-Whitney U test, whereas categorical variables were compared using the chi-squared test or the Fisher exact test (for expected values less than 5). A binary logistic regression for continuous and categorical variables was also performed. Significance was set at p<0.05. Univariate and multivariate analyses were performed. Results Multivariate statistical analysis and clinical interpretation of data identified four factors more associated with a diagnosis of NSTI than serious cellulitis or abscess: elevated lactate on hospital presentation, a patient-reported history of fever, male gender, and homelessness.  Conclusions In patients with upper extremity infections, the clinical presentation of NSTI and serious cellulitis or abscess may appear similar. In this retrospective cohort of patients matched with LRINEC scores, elevated lactate, subjective fever, male gender, and homelessness were significantly associated with NSTI rather than serious cellulitis or abscess.

Google trends as a tool for evaluating public interest in total knee arthroplasty and total hip arthroplasty.

BACKGROUND AND AIMS: There are approximately 1 million total knee arthroplasty (TKA) and total hip arthroplasty (THA) procedures performed annually in the United States. With this number projected to increase, it is vital for orthopedic surgeons and health-care systems to properly anticipate healthcare utilization related to TKA and THA. Google Trends (GT) is a free, open source tool that provides customizable analysis of search terms entered into the Google search engine. We aim to explore the relationship between public interest in TKA and THA as determined by GT data and volume of TKA and THA procedures. METHODS: GT data were compiled for ten search terms related to TKA and ten search terms related to THA from January 2009 to December 2017. Annual case volumes for TKA/THA procedures were obtained from the Healthcare Cost and Utilization Project National Inpatient Sample from 2009 to 2017. Trend analysis was performed using univariate linear regression of GT data and TKA/THA case volumes. RESULTS: There was a statistically significant positive correlation between GT data and procedure volume for 14 of the 20 search terms studied. Seven TKA-related search terms with a positive correlation to procedure volumes include "total knee replacement," "knee replacement," "knee osteoarthritis," "knee ache," "knee swelling," "knee stiffness," and "chronic knee pain." Seven THA-related search terms with a positive correlation to procedure volumes include "hip arthroplasty," "total hip replacement," "hip replacement," "hip osteoarthritis," "hip ache," "hip swelling," and "chronic hip pain." CONCLUSION: GT may provide a high utility as a convenient and informative data set for orthopedic surgeons to analyze public interest in TKA and THA procedures. The data provided by GT have the potential to provide real-time, actionable information that may help surgeons and health systems to characterize public interest in TKA and THA and to best identify and address patient needs. RELEVANCE FOR PATIENTS: The GT tool can be used to measure public interest in TKA/THA, which can inform physician expectations for the patient encounter and lead to the creation of decision aids that better inform the public about the risks and benefits of TKA/THA.

The impact of monthly campaigns and other high-profile media coverage on public interest in 13 malignancies: a Google Trends analysis.

It is estimated that more than 600,000 people will die of cancer in the United States in 2020. Annual cancer diagnoses throughout the country are expected to rise in the coming years, which may further strain the American healthcare system. As such, it is vital that public health campaigns intended to reduce cancer morbidity and mortality are successful. Monthly cancer awareness campaigns have been used in the past to raise awareness and funding for various malignancies. One notable example is the 'Pink October' campaign to raise awareness for breast cancer. There has been limited study, however, on the effectiveness of cancer awareness campaigns for other cancers such as colorectal cancer, prostate cancer and cervical cancer. High-profile media coverage of celebrity cancer diagnoses and/or cancer-related deaths is another method by which knowledge of common cancers is dispersed to the public. In this study, we evaluate the impact of monthly cancer awareness campaigns as well as celebrity cancer diagnoses and/or deaths on Internet search traffic regarding various malignancies. We used the Google Trends database to evaluate public interest in 13 different cancers (and their respective cancer screening methods, when applicable) from January 2010 to June 2020. Public interest in 6 of 13 cancers (cervical cancer, colorectal cancer, skin cancer, ovarian cancer, breast cancer and lung cancer) was significantly higher in their respective awareness months when compared to the rest of the year. Furthermore, peak public interest for 9 of 13 cancers was associated with a media-event such as a monthly awareness campaign or celebrity diagnoses and/or death. Our findings illustrate the important role that the media plays in facilitating public interest in common cancers and their screening methods. Cancer awareness months can serve as an effective tool to increase Internet search traffic regarding a given malignancy. In the future, public health agencies can attempt to utilise increased search traffic to better educate the public, raise funds and improve enrolment in cancer screening programmes that reduce cancer morbidity and mortality.

Invasive penile squamous cell carcinoma.

Invasive penile squamous cell carcinoma (PSCC) is a rare malignancy with considerable morbidity and mortality. Because a delayed or incorrect diagnosis can have a devastating outcome, physicians should maintain a high index of clinical suspicion for PSCC in patients presenting with penile lesions, particularly in young or middle-aged patients in whom presentation of PSCC is uncommon. We report the case of a 27-year-old circumcised man who presented with invasive PSCC following a diagnosis of condyloma 8 years prior. The patient underwent robotic laparoscopic bilateral pelvis and inguinal lymph node dissection and re-excision of the primary PSCC, with one of 15 superficial right inguinal lymph nodes testing positive for squamous cell carcinoma. Given the patient's single-node positivity as well as the risks associated with chemotherapy with respect to his concurrent medical conditions, close follow-up with repeat imaging was planned following surgery. This case highlights the importance of biopsy in any lesion recalcitrant to conventional treatment modalities regardless of age. Early detection and treatment of PSCC can prevent organ dysfunction, loss of organ, or even death.

Investigation of the First Seven Reported Cases of Candida auris, a Globally Emerging Invasive, Multidrug-Resistant Fungus-United States, May 2013-August 2016.

November 11, 2016/65(44);1234-1237. What is already known about this topic? Candida auris is an emerging pathogenic fungus that has been reported from at least a dozen countries on four continents during 2009-2015. The organism is difficult to identify using traditional biochemical methods, some isolates have been found to be resistant to all three major classes of antifungal medications, and C. auris has caused health care-associated outbreaks. What is added by this report? This is the first description of C. auris cases in the United States. C. auris appears to have emerged in the United States only in the last few years, and U.S. isolates are related to isolates from South America and South Asia. Evidence from U.S. case investigations suggests likely transmission of the organism occurred in health care settings. What are the implications for public health practice? It is important that U.S. laboratories accurately identify C. auris and for health care facilities to implement recommended infection control practices to prevent the spread of C. auris. Local and state health departments and CDC should be notified of possible cases of C. auris and of isolates of C. haemulonii and Candida spp. that cannot be identified after routine testing.

The use of Mohs micrographic surgery (MMS) for melanoma in situ (MIS) of the trunk and proximal extremities.

BACKGROUND: Evaluation of the entire surgical margin results in high rates of complete excision, low local recurrence rates, and maximal tissue conservation. Although well recognized for melanoma of the head and neck, few studies have focused exclusively on the trunk and proximal extremities. OBJECTIVE: We sought to evaluate the efficacy of Mohs micrographic surgery for melanoma in situ (MIS) of the trunk and proximal extremities, and determine adequate excision margins for MIS when total margin evaluation is not used. METHODS: Long-term outcomes in 882 cases of MIS treated with Mohs micrographic surgery were analyzed and compared with historical controls. Rates of complete excision were determined for increasing surgical margin intervals. RESULTS: One local recurrence occurred in our cohort (0.1%). Only 83% of MIS were excised with a 6-mm margin. Margins of 9 mm were needed to excise 97% of MIS, statistically equivalent to thin melanomas. LIMITATIONS: We used a nonrandomized, single-institution, retrospective design. CONCLUSION: Mohs micrographic surgery may cure the 17% of MIS that exceed traditional excision margins of 5 mm and is a valuable option for these patients. Surgical margins of at least 0.9 cm should be considered for MIS of the trunk and extremities when total margin evaluation is not used.

The Paramidline Forehead Flap: A Clinical and Microanatomic Study.

BACKGROUND: The traditional paramedian forehead flap (FHF) is an axial flap based on the supratrochlear artery (STA). Doppler examination is often used to ensure inclusion of the STA within the flap. The authors hypothesized that a FHF pedicle design could be simplified to extend from the midline of the glabella to 1.2 cm lateral to the midline without compromising outcomes. OBJECTIVE: To compare clinical outcomes and vessel densities of 2 FHF designs. METHODS: Two FHF designs were used to repair fifty nasal defects. One was based on Doppler identification of the STA; the other on clinical measurements from the glabellar midline (with no effort to identify the STA). Clinical outcomes, complication rates, and pedicle vasculature of both groups were compared. RESULTS: There was no significant difference in flap survival or complication rate. Forehead flaps designed on the paramidline glabella had more arteries within their pedicles than Doppler-based FHFs (p < .05). Small arteries predominated, whereas larger arteries were infrequent in both groups. Size and number of arteries were not related to flap survival. CONCLUSION: A paramidline FHF has equivalent clinical outcomes as a flap based on the STA. A simple and reproducible design of the FHF using only surface landmarks is described.

Nodular amyloidosis: clinical recognition of an unusual entity.

Nodular amyloidosis is a protein deposition disorder that is important to recognize in the clinical setting. Identification and differentiation from primary systemic amyloidosis, which has an identical cutaneous presentation, but serious systemic implications, is of particular significance. Our case report highlights two patients who presented with isolated involvement of the plantar surface and ungual phalanges, each with concomitant tinea pedis. Recognition and diagnosis of cutaneous amyloidosis enables discrimination from systemic disease, and if found, prompt institution of appropriate treatment.

The proximal tubule and albuminuria: really!

Recent data highlight the role of the proximal tubule (PT) in reabsorbing, processing, and transcytosing urinary albumin from the glomerular filtrate. Innovative techniques and approaches have provided exciting insights into these processes, and numerous investigators have shown that selective PT cell defects lead to significant albuminuria, even reaching nephrotic range in animal models. Thus, the mechanisms of albumin reabsorption and transcytosis are undergoing intense study. Working in concert with megalin and cubilin, a nonselective multireceptor complex that predominantly directs proteins for lysosomal degradation, the neonatal Fc receptor (FcRn) located at the brush border of the apical membrane has been implicated as the "receptor" mediating albumin transcytosis. The FcRn pathway facilitates reabsorption and mediates transcytosis by its pH-dependent binding affinity in endosomal compartments. This also allows for selective albumin sorting within the PT cell. This reclamation pathway minimizes urinary losses and catabolism of albumin, thus prolonging its serum half-life. It may also serve as a molecular sorter to preserve and reclaim normal albumin while allowing "altered" albumin to be catabolized via lysosomal pathways. Here, we critically review the data supporting this novel mechanism.

Evaluation of risk factors for vancomycin-resistant Enterococcus bacteremia among previously colonized hematopoietic stem cell transplant patients.

BACKGROUND: Hematopoietic stem cell transplantation (HSCT) recipients colonized with vancomycin-resistant Enterococcus (VRE) may have an increased risk of developing VRE bacteremia. Identification of risk factors for the development of subsequent VRE bacteremia among colonized HSCT recipients is necessary to predict which patients may benefit the most from receiving anti-VRE antibiotic therapy as part of an initial antimicrobial regimen when gram-positive bacteremia is suspected. METHODS: This study was a retrospective chart review conducted from May 2008 to May 2011. Adult HSCT patients admitted to the hospital found to have positive VRE surveillance cultures were included. A multivariate analysis was completed to identify risk factors for the development of VRE bacteremia in the study population. RESULTS: Of 152 patients, 19 (13%) patients developed subsequent VRE bacteremia. Risk factors identified for patients with current VRE colonization for VRE bacteremia were the utilization of vancomycin subsequent to VRE surveillance culture positivity (P = 0.017), prolonged duration of neutropenia (P = 0.001), immunosuppression (P < 0.001), and timing of first VRE surveillance screen positivity at week 1 (P = 0.005). A history of VRE colonization on a prior admission was not an independent risk factor for bacteremia in HSCT patients (P = 1.0). HSCT patients with VRE bacteremia had a 30-day all-cause inpatient mortality rate of 29% (P = 0.001). CONCLUSION: HSCT patients receiving immunosuppressive therapy, who have been exposed to vancomycin subsequent to surveillance culture positivity, have had prolonged neutropenia of >30 days, or first surveillance culture positive at week 1 of admission are potential candidates for early implementation of anti-VRE therapy when a gram-positive bacteremia is suspected.

Evaluating the effectiveness of cognitive-behavioral teletherapy in depressed adults.

Telephone psychotherapy is an emerging form of delivery of care that has recently demonstrated utility and efficacy for adult depression when provided as an adjunct to antidepressant treatment in primary care trials. This study constitutes one of the initial evaluations of cognitive behavioral therapy-telephone treatment (CBT-TT) as a stand-alone treatment for adult depression in specialty care. Thirty adults initiating psychotherapy for depression at a mental health clinic participated in the trial. The majority of participants (69%) were very satisfied with the 8-session CBT-TT, reduction in depression severity was significant over 3 and 6 months, and 42% of participants were considered recovered at termination. These outcomes closely parallel the findings from an earlier primary care trial, despite specialty care participants beginning treatment with more severe depression and without adjunctive antidepressant medication. These findings suggest that CBT-TT for adult depression is feasible and has potential as a stand-alone treatment. Implementation of this telephone-based delivery approach in primary and specialty care settings is discussed.

Urticarial vasculitis: a unique presentation.

Urticarial vasculitis is a relatively rare diagnosis in a patient presenting with urticaria. The process is classically described as a generalized eruption, painful more so than pruritic, lasting longer than 24 hours. Two forms of urticarial vasculitis have been described: ahypocomplementemic form more commonly associated with systemic disease, and a normocomplementemic form that is generally limited to the skin. We report on a uniquely distributed vasculitic eruption restricted mainly to the anterior belt line area in a patient presenting with urticaria and intense pruritus. Urticarial vasculitis as a unique entity is reviewed along with its clinical and histopathologic presentation and the pharmacologic agents used for treatment.