Immune checkpoint inhibitors for POLE or POLD1 proofreading-deficient metastatic colorectal cancer.

BACKGROUND: POLE and POLD1 proofreading deficiency (POLE/D1pd) define a rare subtype of ultramutated metastatic colorectal cancer (mCRC; over 100 mut/Mb). Disease-specific data about the activity and efficacy of immune checkpoint inhibitors (ICIs) in POLE/D1pd mCRC are lacking and it is unknown whether outcomes may be different from mismatch repair-deficient (dMMR)/microsatellite instability-high (MSI-H) mCRCs treated with ICIs. PATIENTS AND METHODS: In this global study, we collected 27 patients with mCRC harboring POLE/D1 mutations leading to proofreading deficiency and treated with anti-programmed cell death-ligand 1 alone +/- anti-cytotoxic T-lymphocyte antigen-4 agents. We collected clinicopathological and genomic characteristics, response, and survival outcomes after ICIs of POLE/D1pd mCRC and compared them with a cohort of 610 dMMR/MSI-H mCRC patients treated with ICIs. Further genomic analyses were carried out in an independent cohort of 7241 CRCs to define POLE and POLD1pd molecular profiles and mutational signatures. RESULTS: POLE/D1pd was associated with younger age, male sex, fewer RAS/BRAF driver mutations, and predominance of right-sided colon cancers. Patients with POLE/D1pd mCRC showed a significantly higher overall response rate (ORR) compared to dMMR/MSI-H mCRC (89% versus 54%; P = 0.01). After a median follow-up of 24.9 months (interquartile range: 11.3-43.0 months), patients with POLE/D1pd showed a significantly superior progression-free survival (PFS) compared to dMMR/MSI-H mCRC [hazard ratio (HR) = 0.24, 95% confidence interval (CI) 0.08-0.74, P = 0.01] and superior overall survival (OS) (HR = 0.38, 95% CI 0.12-1.18, P = 0.09). In multivariable analyses including the type of DNA repair defect, POLE/D1pd was associated with significantly improved PFS (HR = 0.17, 95% CI 0.04-0.69, P = 0.013) and OS (HR = 0.24, 95% CI 0.06-0.98, P = 0.047). Molecular profiling showed that POLE/D1pd tumors have higher tumor mutational burden (TMB). Responses were observed in both subtypes and were associated with the intensity of POLE/D1pd signature. CONCLUSIONS: Patients with POLE/D1pd mCRC showed more favorable outcomes compared to dMMR/MSI-H mCRC to treatment with ICIs in terms of tumor response and survival.

"Nuclear FGF receptor-1 and CREB binding protein: an integrative signaling module".

In this review we summarize the current understanding of a novel integrative function of Fibroblast Growth Factor Receptor-1 (FGFR1) and its partner CREB Binding Protein (CBP) acting as a nuclear regulatory complex. Nuclear FGFR1 and CBP interact with and regulate numerous genes on various chromosomes. FGFR1 dynamic oscillatory interactions with chromatin and with specific genes, underwrites gene regulation mediated by diverse developmental signals. Integrative Nuclear FGFR1 Signaling (INFS) effects the differentiation of stem cells and neural progenitor cells via the gene-controlling Feed-Forward-And-Gate mechanism. Nuclear accumulation of FGFR1 occurs in numerous cell types and disruption of INFS may play an important role in developmental disorders such as schizophrenia, and in metastatic diseases such as cancer. Enhancement of INFS may be used to coordinate the gene regulation needed to activate cell differentiation for regenerative purposes or to provide interruption of cancer stem cell proliferation.

HPV-positive clinically advanced squamous cell carcinoma of the urinary bladder (aBSCC): A comprehensive genomic profiling (CGP) study.

BACKGROUND: Advanced bladder squamous cell carcinoma (aBSCC) is an uncommon form of urinary bladder malignancy when compared with the much higher urothelial carcinoma incidence. We studied the genomic alteration (GA) landscape in a series of aBSCC based on the association with human papilloma virus (HPV) to determine if differences in GA would be observed between the positive and negative groups. METHODS: Using a hybrid capture-based FDA-approved CGP assay, a series of 171 aBSCC were sequenced to evaluate all classes of GA. Tumor mutational burden (TMB) was determined on up to 1.1 Mbp of sequenced DNA and microsatellite instability (MSI) was determined on up to 114 loci. Programmed cell death ligand -1 (PD-L1) expression was determined by IHC (Dako 22C3) with negative expression when PD-L1 was 0, lower expression of positivity set at 1 to 49%, and higher expression set at ≥50% expression. RESULTS: Overall, 11 (6.4%) of the aBSCC were found to harbor HPV sequences (10 HPV16 and 1 HPV 11). HPV+ status was identified slightly more often in women (NS) and in younger patients (P = 0.04); 2 female patients with aBSCC had a prior history of SCC including 1 anal SCC and 1 vaginal SCC. HPV+ aBSCC had fewer GA/tumor (P < 0.0001), more inactivating mutations in RB1 (P = 0.032), and fewer inactivating GA in CDKN2A (P < 0.0001), CDKN2B (P = 0.05), TERT promoter (P = 0.0004) and TP53 (P < 0.0001). GA in genes associated with urothelial carcinoma including FGFR2 and FGFR3 were similar in both HPV+ and HPV- aBSCC groups. MTAP loss (homozygous deletion) which has emerged as a biomarker for PRMT5 inhibitor-based clinical trials was not identified in any of the 11 HPV+ aBSCC cases, which was significantly lower than the 28% positive frequency of MTAP loss in the HPV- aBSCC group (P < 0.0001). MTOR and PIK3CA pathway GA were not significantly different in the 2 groups. Putative biomarkers associated with immunotherapy (IO) response, including MSI and TMB status, were also similar in the 2 groups. PD-L1 expression data was available for a subset of both HPV+ and HPV- cases and showed high frequencies of positive staining which was not different in the 2 groups. CONCLUSIONS: HPV+ aBSCC tends to occur more often in younger patients. As reported in other HPV-associated squamous cell carcinomas, HPV+ aBSCC demonstrates significantly reduced frequencies of inactivating mutations in cell cycle regulatory genes with similar GA in MTOR and PIK3CA pathways. The implication of HPV in the pathogenesis of bladder cancer remains unknown but warrants further exploration and clinical validation.

Healthcare personnel intestinal colonization with multidrug-resistant organisms.

OBJECTIVES: This study aims to assess the association between patient contact and intestinal carriage of multidrug-resistant organisms (MDRO) by sampling healthcare personnel (HCP) and staff without patient contact. METHODS: For this observational study, we recruited 400 HCP who worked in our 200-bed research hospital and 400 individuals without patient contact between November 2013 and February 2015. Participants submitted two self-collected perirectal swabs and a questionnaire. Swabs were processed for multidrug-resistant Gram-negative bacteria and vancomycin-resistant enterococci (VRE). Questionnaires explored occupational and personal risk factors for MDRO carriage. RESULTS: Among 800 participants, 94.4% (755/800) submitted at least one swab, and 91.4% (731/800) also submitted questionnaires. Extended spectrum ß-lactamase-producing organisms were recovered from 3.4% (26/755) of participants, and only one carbapenemase-producing organism was recovered. No VRE were detected. The potential exposure of 68.9% (250/363) of HCP who reported caring for MDRO-colonized patients did not result in a rate of MDRO carriage among HCP (4.0%; 15/379) significantly higher than that of staff without patient contact (3.2%; 12/376; p 0.55). CONCLUSIONS: This is the largest US study of HCP intestinal MDRO carriage. The low colonization rate is probably reflective of local community background rates, suggesting that HCP intestinal colonization plays a minor role in nosocomial spread of MDROs in a non-outbreak setting. TRIAL REGISTRATION: clinicaltrials.gov Identifier: NCT01952158.

Implementation of a pharmacogenomics consult service to support the INGENIOUS trial.

Hospital systems increasingly utilize pharmacogenomic testing to inform clinical prescribing. Successful implementation efforts have been modeled at many academic centers. In contrast, this report provides insights into the formation of a pharmacogenomics consultation service at a safety-net hospital, which predominantly serves low-income, uninsured, and vulnerable populations. The report describes the INdiana GENomics Implementation: an Opportunity for the UnderServed (INGENIOUS) trial and addresses concerns of adjudication, credentialing, and funding.

Clinical Pharmacogenetics Implementation Consortium (CPIC) Guidelines for CYP3A5 Genotype and Tacrolimus Dosing.

Tacrolimus is the mainstay immunosuppressant drug used after solid organ and hematopoietic stem cell transplantation. Individuals who express CYP3A5 (extensive and intermediate metabolizers) generally have decreased dose-adjusted trough concentrations of tacrolimus as compared with those who are CYP3A5 nonexpressers (poor metabolizers), possibly delaying achievement of target blood concentrations. We summarize evidence from the published literature supporting this association and provide dosing recommendations for tacrolimus based on CYP3A5 genotype when known (updates at www.pharmgkb.org).

Changes in the extracellular matrix of the autogenous patellar tendon graft after posterior cruciate ligament reconstruction: a biochemical study in sheep.

The left knee joints of female German sheep were operated, replacing the posterior cruciate ligament by the central third of the patellar tendon. After 2, 6, 16, 26 and 52 weeks the graft of the operated leg as well as the contralateral central third of the patellar tendon and the posterior cruciate ligament were dissected and used for biochemical analysis. The total glycosaminoglycan content in grafts increases within the first year up to 5 fold and is higher than that of the patellar tendon, but is lower than that of the cruciate ligament. The distribution pattern of glycosaminoglycans differ in the posterior cruciate ligament and the patellar tendon. In cruciate ligament the main components are chondroitin sulfate (76%) and hyaluronan (15%). In the patellar tendon a higher portion of dermatan sulfate (approx. 16%) was found, next to 52% chondroitin sulfate and 22% hyaluronan. During the examination time the grafts show changes in the concentration and the distribution pattern of glycosaminoglycans. The chondroitin sulfate content increases during the experimental period from 1.4 +/- 1.2 mumol/g dry weight (d.w.) to 8.7 +/- 2.9 mumol/g d.w. After 1 year the chondroitin sulfate content in the graft does not differ significantly from that of the cruciate ligament. In the grafts the concentration of chondroitin (non sulfated) increases after 6 weeks up to 1.3 +/- 0.6 mumol/g d.w. in comparison to the value after 2 weeks (0.2 +/- 0.1 mumol/g d.w.) and also in the other groups (16, 26 and 52 weeks) it remains significantly increased. After 1 year the dermatan sulfate content in the graft has increased up to the fifth-fold compared to the value after 2 weeks and is higher than in the patellar tendon and in the cruciate ligament. In graft the hyaluronan content (1.0 +/- 0.4 mumol/g d.w. does not differ significantly in the groups 2, 6, 16, 26 and 52 weeks after operation, but in all five groups it is lower than in the cruciate ligament (2.4 +/- 1.0 mumol/g d.w.). Dermatan and heparan sulfate are not or only little detected in all three tissues. The distribution pattern of glycosaminoglycans in graft shows chondroitin sulfate and dermatan sulfate being the major parts of glycosaminoglycans with a slight increase of these components in the different groups during the experimental period. Hyaluronan makes up to 24 +/- 5% of the whole content of glycosaminoglycans after 2 weeks and decrease to 8 +/- 1% after 52 weeks.(ABSTRACT TRUNCATED AT 400 WORDS)

Histochemical aspects of the proteoglycans of patellar tendon autografts used to replace the posterior cruciate ligament.

In the female German black-faced sheep the posterior cruciate ligament was replaced by a free patellar tendon autograft and after 2, 6, 16, 26 and 52 weeks tissue samples of the graft's center (axial region far from bones) were removed for histochemistry and electron microscopy. To localize the proteoglycans Alcian Blue and 0.3 M MgCl2 were added to the fixative solution. The distribution of the proteoglycans in the graft was compared to that of a normal patellar tendon and of a normal posterior cruciate ligament. In the patellar tendon spindle-shaped cells predominated and proteoglycans appeared as short filaments at regular intervals between the collagen fibrils. In the posterior cruciate ligament chondroid cells and long filaments in a net-work-like arrangement were seen. In the patellar tendon autografts short interfibrillar filaments prevailed after 2, 6 and 16 weeks. After 26 weeks and particularly after 52 weeks long filaments also appeared. Digestion with Chondroitinase ABC, AC and Hyaluronidase suggested that the short filaments were PGs containing dermatan sulfate. In grafts, in the early phases the fibroblasts predominated, while in the late phases mainly chondroid cells were observed. The grafts showed aspects of the normal posterior cruciate ligament. However, differences remained, for example the thin collagen fibrils, which could represent one of the reasons for a secondary graft failure.

Localization of carbonic anhydrase IV in rat and human heart muscle.

We investigated carbonic anhydrase IV (CA IV) in rat and human heart with immunohistochemical methods by both light and electron microscopy. In cryosections that were incubated with anti-CA IV/FITC, the capillaries showed a strong reaction for CA IV. In paraffin and semithin sections treated with anti-CA IV/ABC (avidin-biotin-peroxidase complex) blood vessels, capillaries, and sarcolemma (SL) were positively stained. By staining ultrathin sections with anti-CA IV/immunogold, CA IV could also be demonstrated at the latter two locations, including the specialized sarcolemmal structures intercalated discs, and T-tubules. In addition, by this method CA IV was seen to be associated with the sarcoplasmic reticulum (SR). The absence of immunostaining in SR and/or SL with some techniques probably indicates a problem of accessibility of the antigenic sites. In line with the immunohistochemical results, CA IV mRNA expression was visualized in both endothelial and muscle cells by in situ hybridization histochemistry.

Action of FMRFamide-like peptides on porcine gastrointestinal motility in vitro.

Mechanical activity was recorded in circular and longitudinal smooth muscle preparations isolated from extensive regions of the porcine gastrointestinal tract in response to the FMRFamide-like neuropeptides F8Famide and A18Famide. In all preparations, the peptides were about equipotent in producing phasic contractions or enhancing spontaneous activity. The most prominent responses were observed in jejunal longitudinal strips which were on the average 91% (+/- 4% SEM, n = 15; 10(-6) M) of the histamine (10(-5) M) responses. The peptide-induced phasic activity was completely abolished by nifedipine but was unaffected by tetrodotoxin, atropine, phentolamine, yohimbine, phenoxybenzamine, propranolol, methysergide, cimetidine, indomethacin, levallorphane or naloxone. Both peptides enhanced acetylcholine-induced contractions. However, bovine ileum and guinea-pig taenia coli was not affected by these peptides. The results indicate that F8F- and A18F-amide contract porcine gastrointestinal smooth muscle by acting directly via non-opioid receptors on L-type calcium channels. In addition an increase of the sensitivity to cholinergic stimulation occurs.

Large-scale homogeneous molecular templates for femtosecond time-resolved studies of the guest-host interaction.

Self-assembled monolayer films based on iodobenzoyloxy-functionalized resorc[4]arenes were prepared on gold substrates to serve as model systems for future time-resolved studies of molecular recognition, a mechanism of outstanding importance in bioorganic systems. The film properties were tested using X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM) and imaging ellipsometry. An apparatus for time-resolved electron spectroscopy utilizing femtosecond soft X-ray pulses is capable of detecting iodine core-level photolines and the photoinduced dissociation after ultraviolet illumination. The developed technique holds promise for tracking the temporal evolution of chemical shifts of atomic markers as local probes for the dynamics of the guest-host interaction.

Effect of brimonidine tartrate ophthalmic solution 0.2% on pupil size in normal eyes under different luminance conditions.

PURPOSE: To evaluate the effect of brimonidine tartrate ophthalmic solution 0.2% (Alphagan) on pupil size in normal eyes. Three luminance conditions were used to assess the potential use of brimonidine in postoperative refractive patients who experience nighttime vision problems related to large pupil size. SETTING: McDonald Eye Associates, Fayetteville, Arkansas, USA. METHODS: Pupil size was measured in 16 eyes of 16 participants with the Colvard pupillometer under 3 luminance conditions. One drop of brimonidine 0.2% was administered to each patient. Pupil size was then measured using the same technique 30 minutes and 4 and 6 hours after drop administration. RESULTS: Under scotopic conditions, 100% of the pupils showed significant miosis at 30 minutes (P <.05). The effect continued in all eyes for 4 hours. At 6 hours, a miotic effect was still present in 81.3%. However, under photopic luminance, there was no significant effect on pupil size in all 16 eyes (P >.05). The pupil size in 5 eyes (31.2%) was not affected at 30 minutes or 4 or 6 hours. At 6 hours, 15 eyes (93.8%) had returned to their preinstillation size. CONCLUSION: Brimonidine tartrate 0.2% had a significant effect in decreasing pupil size under scotopic conditions. The results indicate that the drug can decrease night-vision difficulties such as halos, star bursts, glare, and monocular diplopia in postoperative refractive patients.

The relationship of mechanical properties to morphology in patellar tendon autografts after posterior cruciate ligament replacement in sheep.

In a sheep model the posterior cruciate ligament (PCL) was replaced by a patellar tendon autograft (PTAG) using the central one-third of the ipsilateral patellar tendon (PT). The sheep were sacrificed at 16, 26, 52 and 104 weeks postoperation. The PTAG, and, as controls, the contralateral PCL and PT were harvested. These were examined using biomechanical testing as well as light and transmission electron microscopy, including immunohistological techniques. The material properties (maximum stress, elastic modulus) were compared to the morphological features. The cellular distribution, the distribution of glycosaminoglycans (GAGs), the collagen fibril diameter and the occurrence of Type III collagen were studied. Prior to transplantation, the PTAG was shown to be superior in maximum stress (57.2 +/- 5.5 MPa vs 41.3 +/- 1.9 MPa) and elastic modulus (368.8 +/- 49.3 MPa vs 172.3 +/- 14.6 MPa) to the PCL. The early decline in material properties of the PTAG (maximum stress 22% and elastic modulus 42% of the control) after free grafting paralleled a cell- and capillary-rich PTAG tissue with remnants of necrosis and a poorly organized extracellular matrix. Two years after implantation, with progressive alignment of the tissue matrix, maximum stress and elastic modulus acquired approximately 60 and 70% of the control, respectively. However, there was also an evidence of degenerative changes characterized by acellular areas, loss of the normal bundling pattern of collagen fibers and abnormal accumulation of GAGs. Ultrastructurally, there was a predominant shift to thin collagen fibrils in the PTAG compared to PCL and PT, both consisting of thick and thin collagen fibrils. Thin fibrils were demonstrated to be, in part, split thick fibrils as well as newly formed fibrils. Most of these thin fibrils revealed a positive reaction with antibodies to Type III collagen.

Alterations of glycosaminoglycans during patellar tendon autograft healing after posterior cruciate ligament replacement. A biochemical study in a sheep model.

In each of 30 skeletally mature sheep, the posterior cruciate ligament was replaced in one knee by a free patellar tendon autograft using the central third of the ipsilateral patellar tendon. The healing autograft was compared with the contralateral posterior cruciate ligament and the patellar tendons and posterior cruciate ligaments of nonoperated animals. The content of glycosaminoglycans, chondroitin sulfate disaccharides, and dermatan sulfate disaccharides was assessed biochemically at six periods during the 2 years after surgery. The total glycosaminoglycans and chondroitin sulfate disaccharides in the native posterior cruciate ligament was threefold that in the native patellar tendon. In contrast, the amount of dermatan sulfate disaccharides was similar in both the native tendon and native ligament. In the autograft, glycosaminoglycans and chondroitin sulfate disaccharides increased significantly to about 144% and 172%, respectively, of the contralateral posterior cruciate ligament at Week 104. The dermatan sulfate disaccharides in the autograft also showed a significant increase up to Week 26, followed by a remarkable but not significant decrease until the end of the study. In the contralateral posterior cruciate ligament, the dermatan sulfate disaccharides increased significantly between Weeks 52 and 104. Thus, the amount of dermatan sulfate disaccharides was similar in both the autograft and the contralateral posterior cruciate ligament after 2 years. This study suggests that the patellar tendon autograft did not completely assume the biochemical properties of the posterior cruciate ligament.

Collagen fibril organization in the patellar tendon autograft after posterior cruciate ligament reconstruction. A quantitative evaluation in a sheep model.

We replaced the posterior cruciate ligament in 30 skeletally mature sheep with a patellar tendon autograft using the central third of the ipsilateral patellar tendon. The healing autograft was compared with the contralateral posterior cruciate ligament and the patellar tendons and posterior cruciate ligaments of nonoperated animals. The collagen fibril diameters were analyzed using transmission electron photomicrographs of fibril cross sections taken at six periods during the 2 years after surgery. The patellar tendon and posterior cruciate ligament were characterized by a broad, nongaussian distribution of collagen fibril diameters. The autografts shifted to a unimodal distribution by an increase of small-diameter collagen fibrils. The frequency of small-diameter fibrils measuring up to 100 nm was 99% after 2 years. At that time, these small-diameter fibrils represented 91.6% of the area covered by collagen fibrils. The mean diameter of the collagen fibrils in the autografts significantly decreased to 45% of the controls at Week 26 and remained at this level until the end of this study. The percentage of area covered by collagen fibrils per 1 micron 2 was 78% of the controls 2 years post-operatively. This study suggests that the patellar tendon autograft could not reproduce the collagen fibril organization of the posterior cruciate ligament. This may be a biologic factor responsible for inconsistent results in posterior cruciate ligament replacement.

Efficacy of a minimal contact version of a multimodal smoking cessation program.

A five-week multimodal smoking cessation program was delivered in two formats: Weekly Contact Group meetings led by graduate student leaders versus a minimal contact By-Mail version consisting of weekly mailings of program materials. By-Mail subjects also had access to a "hotline" number they could call with questions and problems. Of 43 smokers who began the Contact Group program, 37% were abstinent at one-year follow-ups; 41% of the 49 By-Mail subjects were abstinent at one year. These results suggest that cost effective minimal contact cessation programs can benefit the many smokers who are unwilling or unable to visit cessation clinics.

Ultrastructural changes of the patellar tendon as a cruciate ligament substitute (one year and two year results).

In four black-faced sheep, the posterior cruciate ligament was replaced with a free autogenous patellar tendon transplant. Tissue samples from the transplants were investigated by light and electron microscopy 1 year and 2 years after surgery. The normal contralateral posterior cruciate ligament and the normal contralateral patellar tendon were used as controls. The structural differences concerned cells, collagen fibrils, elastic tissue and proteoglycans. Most of the cells of the contralateral patellar tendon were spindle-shaped, whereas those of the transplant were frequently chondroid. In the central region of the transplant as well as in the area far from the bone, cell degenerations, and occasionally hypo- or even acellular zones were found. Measurements of the diameter of collagen fibrils in both contralateral patellar tendon and posterior cruciate ligament showed a more or less pronounced bimodal distribution. A unimodal distribution with mainly thin fibrils (20-60 nm) was demonstrated in the transplant tissue which also revealed some morphological alterations of the collagen fibrils. Thin elastic fibers (microfibrils and amorphous material) were randomly scattered among the collagen fibrils of the control samples, bundles of microfibrils (without amorphous material) characterized the transplant. Staining with Alcian blue in the presence of 0.3 M MgCl2 demonstrated a close relationship between proteoglycans and collagen fibrils as well as elastic components in patellar tendon. This arrangement was lost in the transplant where abundant proteoglycans were revealed which, however, composed a tight irregular network between the collagen fibrils. The results serve as a baseline for understanding the impaired biochemical properties of a free autogenous patellar tendon transplant.

What would Nightingale say?

The letter to Florence Nightingale was written by Bernita Decker as part of a nursing course assignment for our Nurse Educator advisor, Betty Pugh. Florence Nightingale's response to the student was written by Joanne Farley, a Nightingale historian, who recently returned from a trip to England and Ireland to study original Nightingale documents and historic sites.

A comparison of the individualized education plan and the individualized family service plan.

The individualized education plan (IEP) and the individualized family service plan (IFSP) are mandated for children with special needs. Occupational therapists participate in the development of both the IEP and the IFSP. This paper summarizes the similarities and the differences in the mandated components. The components addressed are (a) information about the child's status, (b) information about the family, (c) outcomes for the child and family, (d) intervention services, (e) other services, (f) dates and duration of services, (g) selection of a case manager, and (h) transition plans.