Overcoming a Radical Polarity Mismatch in Strain-Release Pentafluorosulfanylation of [1.1.0]Bicyclobutanes: An Entryway to Sulfone- and Carbonyl-Containing SF5-Cyclobutanes.

The first assortment of achiral pentafluorosulfanylated cyclobutanes (SF5-CBs) are now synthetically accessible through strain-release functionalization of [1.1.0]bicyclobutanes (BCBs) using SF5Cl. Methods for both chloropentafluorosulfanylation and hydropentafluorosulfanylation of sulfone-based BCBs are detailed herein, as well as proof-of-concept that the logic extends to tetrafluoro(aryl)sulfanylation, tetrafluoro(trifluoromethyl)sulfanylation, and three-component pentafluorosulfanylation reactions. The methods presented enable isolation of both syn and anti isomers of SF5-CBs, but we also demonstrate that this innate selectivity can be overridden in chloropentafluorosulfanylation; that is, an anti-stereoselective variant of SF5Cl addition across sulfone-based BCBs can be achieved by using inexpensive copper salt additives. Considering the SF5 group and CBs have been employed individually as nonclassical bioisosteres, structural aspects of these unique SF5-CB "hybrid isosteres" were then contextualized using SC-XRD. From a mechanistic standpoint, chloropentafluorosulfanylation ostensibly proceeds through a curious polarity mismatch addition of electrophilic SF5 radicals to the electrophilic sites of the BCBs. Upon examining carbonyl-containing BCBs, we also observed rare instances whereby radical addition to the 1-position of a BCB occurs. The nature of the key C(sp3)-SF5 bond formation step - among other mechanistic features of the methods we disclose - was investigated experimentally and with DFT calculations. Lastly, we demonstrate compatibility of SF5-CBs with various downstream functionalizations.

Small donor size does not negatively impact outcomes after deceased-donor renal transplantation.

Higher body mass index (BMI) of deceased-donors is associated with poorer outcomes in transplant recipients. The effect of low donor BMI on recipient graft function is not clear. Scientific Registry of Transplant Recipients data on recipients of deceased-donor kidneys from 2000 to 2019 were categorized by donor BMI (donor BMI < 18, 18-27, and >27). Primary outcome was death-censored graft survival. The impact of multiple recipient and donor variables, including low donor BMI and the difference between donor and recipient BMI, was evaluated using a multivariate Cox proportional-hazards model. Low BMI donors (LBD) were more likely to be younger, female, and white (all P < .05). LBD were less likely to be Hispanic, diabetic, or have hypertension (all P < .001). LBD recipients were more likely to be younger and female (both P < .001). Low donor BMI was not significantly associated with recipient graft survival. Donor-recipient BMI difference did not correlate with an increased risk of graft failure. Similar results were obtained when donors were classified using body surface area (BSA). Small donor size in terms of BMI or BSA or a large discrepancy between donor and recipient size should not necessarily preclude transplantation of an otherwise acceptable kidney.

Effects of MAL-PDT, ingenol mebutate and diclofenac plus hyaluronate gel monitored by high-frequency ultrasound and digital dermoscopy in actinic keratosis - a randomized trial.

BACKGROUND: The efficacy for actinic keratosis (AK) clearance of field-directed treatments has been investigated in randomized studies against placebo, but the comparison of results is difficult for several methodological reasons. OBJECTIVES: The present study aims to compare efficacy of MAL-photodynamic therapy (MAL-PDT), ingenol mebutate gel (IMB) and diclofenac plus hyaluronate gel (DHA) on multiple AKs assessing a new performance index of efficacy, the cumulative AK area and evaluating dermoscopical and high-frequency ultrasound (HFUS) changes. METHODS: Patients with ≥5 Olsen II AKs in a 25 cm2 area of the scalp and face were enrolled and randomized to one of the treatment choices. Number of AKs and cumulative area were assessed before and after treatment. Dermoscopy and HFUS were performed on a single AK and surrounding photo-damaged skin in the treatment area. RESULTS: Cumulative AKs area reduced significantly more with PDT compared to other treatment options and with IMB in comparison to DHA. PDT was also the only treatment option that increased at a significant level the dermal density in both target AK and the surrounding skin and decreased significantly the SLEB thickness in the perilesional skin at HFUS. CONCLUSIONS: MAL-PDT is more effective than IMB and DHA for reducing the cumulative AK area which is calculated digitally from 3D pictures and should be the preferred performance index for the evaluation of the efficacy of treatments for AKs, rolling out clinical and dermoscopy evaluation. MAL-PDT improves all HFUS features of chronic photodamages of the dermis of the skin underlying and surrounding the AK spots.

Robotic-assisted Donor Nephrectomy: As Safe as Laparoscopic Donor Nephrectomy.

OBJECTIVES: Robotic-assisted donor nephrectomy has become increasingly popular in recent years. We sought to compare robotic-assisted outcomes to operative outcomes in a historical cohort of laparoscopic donor nephrectomies. MATERIALS AND METHODS: A retrospective review of 58 consecutive donor nephrectomies at a single center by two surgeons from 2015 to 2019 was performed. RESULTS: Robotic-assisted (n = 32) and laparoscopic (n =26) donors were comparable in terms of estimated blood loss (66.4 vs. 62.5 mL; p=0.81) and length of stay (1.6 vs. 1.5 days; p=0.37). The post-operative change in creatinine was not significantly different between the groups (-0.45 vs. -0.45; p=0.97). Warm ischemia time was longer in the robotic group (7.36 vs. 5.15 minutes; p < 0.01). Case duration was significantly longer for robotic-assisted cases (306 vs. 247 minutes; p < 0.01). However, robotic cases were more likely to be right-sided (6/32 (18.8%) vs. 1/26 (3.8%)) and have vascular multiplicity (7/32 (21.9%) vs. 0/26 (0%)). There was one Clavien II or greater complication in the robotic group, and none in the laparoscopic group. CONCLUSIONS: Our data agree with previous reports that robotic-assisted donor nephrectomy is safe and has similar outcomes to a laparoscopic approach. Moreover, cases with multiple renal arteries or veins and right-sided cases were completed safely using the robotic approach. The longer case duration in the robotic group was attributed to the learning curve associated with implementing a new technique, and later robotic operative times approached laparoscopic operative times.

A systematic review of service-learning in medical education: 1998-2012.

UNLABELLED: PHENOMENON: In the United States, the Affordable Care Act has increased the need for community-centered pedagogy for medical education such as service-learning, wherein students connect academic curriculum and reflections to address a community need. Yet heterogeneity among service-learning programs suggests the need for a framework to understand variations among service-learning programs in medical education. APPROACH: A qualitative systematic review of literature on service-learning and medical education was conducted for the period between 1998 and 2012. A two-stage inclusion criteria process resulted in articles (n = 32) on service-learning and Doctor of Medicine or Doctor of Osteopathic Medicine being included for both coding and analysis. Focused and selective coding were employed to identify recurring themes and subthemes from the literature. FINDINGS: The findings of the qualitative thematic analysis of service-learning variation in medical education identified a total of seven themes with subthemes. The themes identified from the analysis were (a) geographic location and setting, (b) program design, (c) funding, (d) participation, (e) program implementation, (f) assessment, and (g) student outcomes. Insights: This systematic review of literature confirmed the existence of program heterogeneity among service-learning program in medical education. However, the findings of this study provide key insights into the nature of service-learning in medical education building a framework for which to organize differences among service-learning programs. A list of recommendations for future areas of inquiry is provided to guide future research.

A randomized clinical trial of photodynamic therapy with methyl aminolaevulinate vs. diclofenac 3% plus hyaluronic acid gel for the treatment of multiple actinic keratoses of the face and scalp.

BACKGROUND: A gel containing diclofenac and hyaluronic acid (DHA) and photodynamic therapy with methyl aminolaevulinate (MAL-PDT) are widely used treatments for actinic keratoses (AKs). OBJECTIVES: The aim of this single-centre, open-label, prospective, nonsponsored, randomized controlled clinical trial was to compare the treatment results and cost-effectiveness of MAL-PDT and DHA. METHODS: Patients with multiple AKs of the face and scalp were randomized to receive MAL-PDT or DHA. After 90 days, the overall complete remission (CR) rate of patients and the CR rate of lesions according to thickness score were assessed, and patients and an investigator scored the cosmetic outcome. In addition, patients scored their overall satisfaction with the treatment. Patients with CR of all lesions were followed up for 12 months. RESULTS: Two hundred patients with a total of 1674 AKs were enrolled. The lesion CR rates at 3 months were 85·9% with MAL-PDT and 51·8% with DHA (P < 0·0001). AKs of all thicknesses were significantly more responsive to MAL-PDT. The patient CR rates at 3 months were 68% with MAL-PDT and 27% with DHA. At the 12-month examination, the number of patients with CR reduced to 37 with MAL-PDT and seven with DHA. Rating of cosmetic outcome was very good or excellent in the vast majority of patients with both treatments. The analysis of cost-effectiveness showed that the costs per patient with CR at 3 months and at 12 months are €566·7 and €1026·2, respectively, with MAL-PDT and €595·2 and €2295·6, respectively, with DHA. CONCLUSIONS: Efficacy, cosmetic outcome and patients' overall satisfaction with MAL-PDT are superior in comparison with DHA. MAL-PDT is more expensive but it is more cost-effective.

Cryotherapy is preferable to ablative CO2 laser for the treatment of isolated actinic keratoses of the face and scalp: a randomized clinical trial.

BACKGROUND: Actinic keratosis (AK) may progress to squamous cell carcinoma. In the case of normal or mildly photodamaged skin, lesion-directed treatments are considered valuable options despite poor published evidence of their therapeutic activity. OBJECTIVES: The aim of this single-centre, open-label, prospective, nonsponsored, randomized, controlled clinical trial was to compare CO2 laser ablation with cryotherapy in the treatment of isolated AKs of the face and scalp. PATIENTS AND METHODS: Patients with isolated (≤ 4) AKs of the face and scalp were randomized to receive CO2 laser ablation or cryotherapy. After 90 days, the overall complete remission (CR) rates of patients and lesions were assessed and correlated with thickness grade. RESULTS: Two hundred patients with a total number of 543 AKs were enrolled. The CR rates of lesions after 3 months were 78·2% with cryotherapy and 72·4% with CO2 laser ablation. Thicker lesions were significantly more responsive to cryotherapy (P = 0·034). Seventy-three patients (71·6%) had CR of all lesions 3 months after cryotherapy and 64 (65·3%) after laser ablation. At 12 months after treatment the number of patients with CR was reduced to 53 with cryotherapy and 14 with laser ablation. CONCLUSIONS: The rate of patients and lesions with CR is similar after 3 months, but more patients remain in stable remission for 12 months after cryotherapy. Cryotherapy is more effective for thick lesions. The cosmetic outcome was good or excellent in almost all patients.

Simple Behavioral Analysis (SimBA) as a platform for explainable machine learning in behavioral neuroscience.

The study of complex behaviors is often challenging when using manual annotation due to the absence of quantifiable behavioral definitions and the subjective nature of behavioral annotation. Integration of supervised machine learning approaches mitigates some of these issues through the inclusion of accessible and explainable model interpretation. To decrease barriers to access, and with an emphasis on accessible model explainability, we developed the open-source Simple Behavioral Analysis (SimBA) platform for behavioral neuroscientists. SimBA introduces several machine learning interpretability tools, including SHapley Additive exPlanation (SHAP) scores, that aid in creating explainable and transparent behavioral classifiers. Here we show how the addition of explainability metrics allows for quantifiable comparisons of aggressive social behavior across research groups and species, reconceptualizing behavior as a sharable reagent and providing an open-source framework. We provide an open-source, graphical user interface (GUI)-driven, well-documented package to facilitate the movement toward improved automation and sharing of behavioral classification tools across laboratories.

Photodynamic therapy for basal cell carcinoma: clinical and pathological determinants of response.

BACKGROUND: Photodynamic therapy (PDT) is increasingly used in the treatment of basal cell carcinoma (BCC). However, scant information is available about the impact of both patient- and lesion-related characteristics on the effectiveness of therapy. Therefore, on the basis of the current data, it is difficult to draw clear-cut indications to use PDT for treatment of BCC in clinical practice. OBJECTIVE: To investigate the clinical and pathological determinants of response of BCC to PDT with methylaminolevulinate (MAL) and red light. METHODS: The clinical and pathological characteristics of 194 BCCs in 135 patients, treated with MAL-PDT, were evaluated. Lesions were treated with MAL-PDT according to established methods and the response was assessed by clinical follow-up of the patients. RESULTS: Complete response to PDT was 62%, with a better response for superficial BCC (95/116, 82%) than nodular BCC (26/78, 33%). When determinants of response were analysed, the nodular type and the location on the limbs emerged as significant clinical predictors of failure. Among the pathological characteristics, the nodular and infiltrative histotypes, as well as ulceration and tumour thickness were associated with a lower response to therapy. Patients' age and gender, as well as the size of the lesions, were not found to be significant predictors. CONCLUSIONS: Optimization of PDT procedure for BCC requires a careful selection of the lesions. In particular, superficial BCCs, preferentially located on the trunk, show the best therapeutic response.

PD-L1 pathway as a novel target in carcinosarcoma of the kidney and renal pelvis.

Carcinosarcoma of the kidney and renal pelvis (CSKP) is a rare and highly-aggressive malignancy characterized by rapid progression and widespread metastases. To date, few studies describe the natural history of the disease. We present a patient placed on pembrolizumab therapy for suspected metastatic colon cancer. The patient was found to have a right renal mass with caval extension on surveillance and ultimately underwent radical surgery revealing carcinosarcoma with positive PD-L1 expression with no evidence of recurrence to date. To our knowledge, this is the first case describing PD-L1 expression in CSKP and presents a novel pathway for future treatment algorithms.

Amygdala-Midbrain Connections Modulate Appetitive and Aversive Learning.

The central amygdala (CeA) orchestrates adaptive responses to emotional events. While CeA substrates for defensive behaviors have been studied extensively, CeA circuits for appetitive behaviors and their relationship to threat-responsive circuits remain poorly defined. Here, we demonstrate that the CeA sends robust inhibitory projections to the lateral substantia nigra (SNL) that contribute to appetitive and aversive learning in mice. CeA→SNL neural responses to appetitive and aversive stimuli were modulated by expectation and magnitude consistent with a population-level salience signal, which was required for Pavlovian conditioned reward-seeking and defensive behaviors. CeA→SNL terminal activation elicited reinforcement when linked to voluntary actions but failed to support Pavlovian associations that rely on incentive value signals. Consistent with a disinhibitory mechanism, CeA inputs preferentially target SNL GABA neurons, and CeA→SNL and SNL dopamine neurons respond similarly to salient stimuli. Collectively, our results suggest that amygdala-nigra interactions represent a previously unappreciated mechanism for influencing emotional behaviors.

Methylaminolaevulinate-based photodynamic therapy of Bowen's disease and squamous cell carcinoma.

BACKGROUND: Photodynamic therapy (PDT) with methylaminolaevulinate (MAL) is an approved noninvasive treatment option for actinic keratosis and Bowen's disease (BD), two precursors of invasive squamous cell carcinoma (SCC). OBJECTIVES: To assess efficacy, prognostic features, tolerability and cosmetic outcome of MAL-PDT for the treatment of BD and SCC. METHODS: In total, 112 biopsy-proven lesions of BD and SCC in 55 subjects were treated in an outpatient setting. MAL cream (160 mg g(-1)) was applied for 3 h prior to illumination with a light-emitting diode source (wavelength range 635 +/- 18 nm; light dose 37 J cm(-2)). A second MAL-PDT session was given 7 days later. Complete response rate at 3 months after the last treatment, recurrence rate at the 24-month follow-up, and cosmetic outcome were recorded. RESULTS: The overall complete response rates were 73.2% at 3 months and 53.6% at 2 years. Clinical thickness, atypia and lesion depth were significant predictors of the response at 3 months when using a univariate analysis (P < 0.001). A multivariate logistic regression model, with robust variance estimation, showed that cell atypia was the only statistically significant independent predictor of the treatment outcome at 3 months. CONCLUSIONS: MAL-PDT may represent a valuable, effective and well tolerated treatment option with good cosmetic outcome for superficial, well-differentiated (Broders' scores I and II) BD and microinvasive SCC. In contrast, its use for superficial SCCs with a microinvasive histological pattern and for nodular, invasive lesions, particularly if poorly differentiated keratinocytes are present (Broders' scores III and IV), should be avoided.

Rapidly reversible hydrophobization: an approach to high first-pass drug extraction.

We have investigated a rapidly reversible hydrophobization of therapeutic agents for improving first-pass uptake in locoregional drug therapy. This approach involves the attachment of a hydrophobic moiety to the drug by highly labile chemical linkages that rapidly hydrolyze upon injection. Hydrophobization drastically enhances cell-membrane association of the prodrug and, consequently, drug uptake, while the rapid lability protects nontargeted tissues from exposure to the highly active agent. Using the membrane-impermeable DNA intercalator propidium iodide, and melphalan, we report results from in vitro cellular internalization and toxicity studies. Additionally, we report in vivo results after a single liver arterial bolus injection, demonstrating both tumor targeting and increased survival in a mouse tumor model.

Initial Experience with the Use of a Robotic Stapler for Robot-Assisted Donor Nephrectomy.

INTRODUCTION: Since the first description of robot-assisted laparoscopic donor nephrectomy for living donor renal transplantation, additional studies have noted outcomes comparable to traditional laparoscopic surgery. Urologists have embraced the technical advantages of the robotic approach, including improved dissection and more accurate graft preservation. With the advent of robotic stapling, we aim to demonstrate the feasibility of the use of robotic stapling for living-donor nephrectomies. MATERIALS AND METHODS: Ten cases of da Vinci® EndoWrist® Stapler use during robot-assisted donor nephrectomy by a single surgeon are reported. Variables examined included patient anatomy, estimated blood loss (EBL), warm ischemia time (WIT), postoperative complications, and graft function at follow-up. The costs of the laparoscopic and robotic staplers at our institution are reported. RESULTS: One right and nine left nephrectomies were performed by a single surgeon. Eight living donors had a single renal artery and vein, while two had multiple renal arteries and renal veins. A single vascular stapler load (45-mm, white cartridge) was used for vessels in all cases. WIT was <6 minutes in each case, and EBL ranged from 13 to 100 mL. Sufficient length on the renal artery and vein was obtained to execute anastomoses in all cases. Only one patient had postoperative complications that were related to anesthesia. Graft function of recipients and postoperative renal function has been adequate to date. The cost of using the robotic stapler was $705 (EndoWrist Stapler), and the cost of the laparoscopic stapler (Endo GIA™ Stapler; Medtronic, Dublin, Ireland) was $494.06. CONCLUSIONS: The use of robotic stapler is feasible in terms of patient safety, technique, postoperative outcomes, and cost in both left- and right-sided living-donor robotic nephrectomies. Further research is needed to confirm the benefit and safety of EndoWrist stapler use in robotic donor nephrectomies compared to laparoscopic stapler use.

HIF2α-Targeted RNAi Therapeutic Inhibits Clear Cell Renal Cell Carcinoma.

Targeted therapy against VEGF and mTOR pathways has been established as the standard-of-care for metastatic clear cell renal cell carcinoma (ccRCC); however, these treatments frequently fail and most patients become refractory requiring subsequent alternative therapeutic options. Therefore, development of innovative and effective treatments is imperative. About 80%-90% of ccRCC tumors express an inactive mutant form of the von Hippel-Lindau protein (pVHL), an E3 ubiquitin ligase that promotes target protein degradation. Strong genetic and experimental evidence supports the correlate that pVHL functional loss leads to the accumulation of the transcription factor hypoxia-inducible factor 2α (HIF2α) and that an overabundance of HIF2α functions as a tumorigenic driver of ccRCC. In this report, we describe an RNAi therapeutic for HIF2α that utilizes a targeting ligand that selectively binds to integrins αvß3 and αvß5 frequently overexpressed in ccRCC. We demonstrate that functional delivery of a HIF2α-specific RNAi trigger resulted in HIF2α gene silencing and subsequent tumor growth inhibition and degeneration in an established orthotopic ccRCC xenograft model. Mol Cancer Ther; 17(1); 140-9. ©2017 AACR.

Genetic immunization for antibody generation in research animals by intravenous delivery of plasmid DNA.

Genetic immunization is an attractive approach to generate antibodies because native proteins are expressed in vivo with normal posttranscriptional modifications, avoiding time-consuming and costly antigen isolation or synthesis. Hydrodynamic tail or limb vein delivery of naked plasmid DNA expression vectors was used to induce antigen-specific antibodies in mice, rats, and rabbits. Both methods allowed the efficient generation of high-titer, antigen-specific antibodies with an overall success rate of Western detectable antibodies of 78% and 92%, respectively. High-titer antibodies were typically present after 3 hydrodynamic tail vein plasmid DNA deliveries, 5 weeks after the initial injection (i.e., prime). For hydrodynamic limb vein plasmid DNA delivery, two deliveries were sufficient to induce high-titer antibody levels. Tail vein delivery was less successful at generating antibodies directed against secreted proteins as compared with limb vein delivery. Material for screening was generated by,transfection of the immunization vector into mammalian cell lines. The cell line (COS-7) that produced the highest level of antigen expression performed best in Western blot analysis screens. In summary, intravenous delivery of antigen-expressing plasmid DNA vectors is an effective genetic immunization method for the induction of antigen-specific antibodies in small and large research animals.

Enhanced activity of hu14.18-IL2 immunocytokine against murine NXS2 neuroblastoma when combined with interleukin 2 therapy.

Established s.c. NXS2 murine neuroblastoma tumors exhibited transient resolution after suboptimal therapy using the hu14.18-IL2 immunocytokine (IC). The hu14.18-IL2 IC is a fusion protein that has linked a molecule of interleukin 2 (IL-2) to the COOH terminus of each of the IgG heavy chains on the humanized anti-GD(2) monoclonal antibody hu14.18. To induce more potent and longer lasting in vivo antitumor effects, we tested hu14.18-IL2 IC in a regimen combining it with constant infusion IL-2 in NXS2 tumor-bearing mice. The addition of the constant infusion IL-2 augmented the antitumor response induced by treatment with the hu14.18-IL2 IC in animals with experimentally induced hepatic metastases and in animals bearing localized s.c. tumors. The combined treatment induced prolonged tumor eradication in most animals bearing s.c. tumors and involved both natural killer cells and T cells. The enhanced ability of this combined treatment to prevent tumor recurrence was not observed when a larger dose of hu14.18-IL2 IC, similar in IL-2 content to the IC plus systemic IL-2 regimen, was tested as single-agent therapy. Animals showing prolonged tumor eradication of established tumors after the combined hu14.18-IL2 plus IL-2 regimen exhibited a protective T-cell-dependent antitumor memory response against NXS2 rechallenge.

The effects of ascorbic acid, vitamin B6, vitamin B12, and folic acid supplementation on the breast milk and maternal nutritional status of low socioeconomic lactating women.

In this double-blind study, the effects of ascorbic acid, vitamin B6, vitamin B12, and folate supplementation on the maternal nutritional status and vitamin content of breast milk in 16 low socioeconomic lactating women were studied. Nine subjects were administered a commercial multivitamin-multimineral supplement and seven were given placebos. Milk samples were collected four times per day at 4-h intervals from 5 to 7 and 43 to 45 days postpartum. Fasting blood samples were taken from all mothers at the end of each milk collection period. Dietary records of all foods consumed were kept from 4 to 7 and 42 to 45 days postpartum. There were no significant differences in milk or plasma levels of ascorbic acid between the unsupplemented and supplemented groups. Both the EGPT index and milk concentration of vitamin B6 were significantly different (p less than 0.01) between the unsupplemented and supplemented groups. The milk concentrations of vitamin B12 increased significantly (p less than 0.05) in the supplemented group, as did the milk concentrations of folate (p less than 0.01). Because of consistently low levels of dietary vitamin B6 and folic acid in this group of low socioeconomic lactating women, either dietary changes or supplements could be necessary to maintain recommended levels of these vitamins in the womens' breast milk.