Trained Immunity and Reactivity of Macrophages and Endothelial Cells.

Innate immune cells can develop exacerbated immunologic response and long-term inflammatory phenotype following brief exposure to endogenous or exogenous insults, which leads to an altered response towards a second challenge after the return to a nonactivated state. This phenomenon is known as trained immunity (TI). TI is not only important for host defense and vaccine response but also for chronic inflammations such as cardiovascular and metabolic diseases such as atherosclerosis. TI can occur in innate immune cells such as monocytes/macrophages, natural killer cells, endothelial cells (ECs), and nonimmune cells, such as fibroblast. In this brief review, we analyze the significance of TI in ECs, which are also considered as innate immune cells in addition to macrophages. TI can be induced by a variety of stimuli, including lipopolysaccharides, BCG (bacillus Calmette-Guerin), and oxLDL (oxidized low-density lipoprotein), which are defined as risk factors for cardiovascular and metabolic diseases. Furthermore, TI in ECs is functional for inflammation effectiveness and transition to chronic inflammation. Rewiring of cellular metabolism of the trained cells takes place during induction of TI, including increased glycolysis, glutaminolysis, increased accumulation of tricarboxylic acid cycle metabolites and acetyl-coenzyme A production, as well as increased mevalonate synthesis. Subsequently, this leads to epigenetic remodeling, resulting in important changes in chromatin architecture that enables increased gene transcription and enhanced proinflammatory immune response. However, TI pathways and inflammatory pathways are separated to ensure memory stays when inflammation undergoes resolution. Additionally, reactive oxygen species play context-dependent roles in TI. Therefore, TI plays significant roles in EC and macrophage pathology and chronic inflammation. However, further characterization of TI in ECs and macrophages would provide novel insights into cardiovascular disease pathogenesis and new therapeutic targets. Graphic Abstract: A graphic abstract is available for this article.

Aberrant Exon 8/8a Splicing by Downregulated PTBP (Polypyrimidine Tract-Binding Protein) 1 Increases CaV1.2 Dihydropyridine Resistance to Attenuate Vasodilation.

OBJECTIVE: Calcium channel blockers, such as dihydropyridines, are commonly used to inhibit enhanced activity of vascular CaV1.2 channels in hypertension. However, patients who are insensitive to such treatments develop calcium channel blocker-resistant hypertension. The function of CaV1.2 channel is diversified by alternative splicing, and the splicing factor PTBP (polypyrimidine tract-binding protein) 1 influences the utilization of mutually exclusive exon 8/8a of the CaV1.2 channel during neuronal development. Nevertheless, whether and how PTBP1 makes a role in the calcium channel blocker sensitivity of vascular CaV1.2 channels, and calcium channel blocker-induced vasodilation remains unknown. Approach and Results: We detected high expression of PTBP1 and, inversely, low expression of exon 8a in CaV1.2 channels (CaV1.2E8a) in rat arteries. In contrast, the opposite expression patterns were observed in brain and heart tissues. In comparison to normotensive rats, the expressions of PTBP1 and CaV1.2E8a channels were dysregulated in mesenteric arteries of hypertensive rats. Notably, PTBP1 expression was significantly downregulated, and CaV1.2E8a channels were aberrantly increased in dihydropyridine-resistant arteries compared with dihydropyridine-sensitive arteries of rats and human. In rat vascular smooth muscle cells, PTBP1 knockdown resulted in shifting of CaV1.2 exon 8 to 8a. Using patch-clamp recordings, we demonstrated a concomitant reduction of sensitivity of CaV1.2 channels to nifedipine, due to the higher expression of CaV1.2E8a isoform. In vascular myography experiments, small interfering RNA-mediated knockdown of PTBP1 attenuated nifedipine-induced vasodilation of rat mesenteric arteries. CONCLUSIONS: PTBP1 finely modulates the sensitivities of CaV1.2 channels to dihydropyridine by shifting the utilization of exon 8/8a and resulting in changes of responses in dihydropyridine-induced vasodilation.

Metabolic Reprogramming in Immune Response and Tissue Inflammation.

Innate and adaptive immunity participate in and regulate numerous human diseases. Increasing evidence implies that metabolic reprogramming mediates immune cell functional changes during immune responses. In this review, we present and discuss our current understanding of metabolic regulation in different immune cells and their subsets in response to pathological stimuli. An interactive biochemical and molecular model was established to characterize metabolic reprogramming and their functional implication in anti-inflammatory, immune resolution, and proinflammatory responses. We summarize 2 major features of metabolic reprogramming in inflammatory stages in innate and adaptive immune cells: (1) energy production and biosynthesis reprogramming, including increased glycolysis and decreased oxidative phosphorylation, to secure faster ATP production and biosynthesis for defense response and damage repair and (2) epigenetic reprogramming, including enhanced histone acetylation and suppressed DNA methylation, due to altered accessibility of acetyl/methyl group donor and metabolite-modulated enzymatic activity. Finally, we discuss current strategies of metabolic and epigenetic therapy in cardiovascular disease and recommend cell-specific metabolic and gene-targeted site-specific epigenetic alterations for future therapies.

Ly6C+ Inflammatory Monocyte Differentiation Partially Mediates Hyperhomocysteinemia-Induced Vascular Dysfunction in Type 2 Diabetic db/db Mice.

OBJECTIVE: Hyperhomocysteinemia (HHcy) is a potent risk factor for diabetic cardiovascular diseases. We have previously reported that hyperhomocysteinemia potentiates type 1 diabetes mellitus-induced inflammatory monocyte differentiation, vascular dysfunction, and atherosclerosis. However, the effects of hyperhomocysteinemia on vascular inflammation in type 2 diabetes mellitus (T2DM) and the underlying mechanism are unknown. Approach and Results: Here, we demonstrate that hyperhomocysteinemia was induced by a high methionine diet in control mice (homocysteine 129 µmol/L), which was further worsened in T2DM db/db mice (homocysteine 180 µmol/L) with aggravated insulin intolerance. Hyperhomocysteinemia potentiated T2DM-induced mononuclear cell, monocyte, inflammatory monocyte (CD11b+Ly6C+), and M1 macrophage differentiation in periphery and aorta, which were rescued by folic acid-based homocysteine-lowering therapy. Moreover, hyperhomocysteinemia exacerbated T2DM-impaired endothelial-dependent aortic relaxation to acetylcholine. Finally, transfusion of bone marrow cells depleted for Ly6C by Ly6c shRNA transduction improved insulin intolerance and endothelial-dependent aortic relaxation in hyperhomocysteinemia+T2DM mice. CONCLUSIONS: Hyperhomocysteinemia potentiated systemic and vessel wall inflammation and vascular dysfunction partially via inflammatory monocyte subset induction in T2DM. Inflammatory monocyte may be a novel therapeutic target for insulin resistance, inflammation, and cardiovascular complications in hyperhomocysteinemia+T2DM.

[HIV infection status and characteristics of non-remunerated blood donors in Hangzhou, 2008-2017].

Objective: To understand HIV infection status and characteristics of non-remunerated blood donors in Hangzhou. Methods: HIV antibody test were conducted for non-remunerated blood donors in Hangzhou and their demographic and epidemiological information were collected from 2008-2017. χ(2) test for trend (liner by liner association chi square test) was used for the comparison of the HIV infection trends in each year. Results: A total of 1 461 129 non-remunerated blood donors were surveyed in Hangzhou during 2008-2017, and 260 blood donors were HIV positive. Most HIV infected blood donors were males (96.5%, 251/260) and aged 18-34 years (72.7%, 189/260). Among 260 HIV positive blood donors, those reporting repeated non-remunerated blood donation accounted for 36.9% (96/260), those reporting homosexual transmission accounted for 53.5% (139/260) and those reporting heterosexual transmission accounted for 44.6% (116/260). The HIV infected persons reporting homosexual behaviors were mainly aged 18-34 years (82.0%, 114/139) and unmarried (71.2%, 99/139). Most HIV infected students reported homosexual transmission (88.4%, 23/26). The crude HIV positive rate was 0.8/10 000-2.5/10 000, the differences in annual HIV positive rate had no significance (trend χ(2)=2.355, P=0.125). The crude HIV positive rate in male blood donors aged 18-24 years increased from 1.1/10 000 in 2008 to 3.7/10 000 in 2017, the difference was significant (trend χ(2)=5.175, P=0.023). Standardized HIV positive rate was 0.9/10 000-2.4/10 000. Conclusions: HIV infection rate was low in non-remunerated blood donors in Hangzhou during 2008-2017. Most HIV infected persons were males and aged 18-34 years. Heterosexual and homosexual contacts were the major transmission routes. The HIV positive rate in males aged 18-24 years showed an increase trend.

[Study on the mechanism of serotype conversion of historical isolates of Shigella flexneri].

Objective: The serotype screening of Shigella flexneri from 1934 to 1965 preserved by the National Center for Medical Culture Collections was carried out, and the molecular characteristics of the serotype conversion strains were studied. Methods: Serotyping of Shigella flexneri in this study was conducted by slide agglutination and multiplex PCR, respectively. The gtrⅡ gene sequence alignment and pulsed field gel electrophoresis typing were performed on the serotype conversion strains. Results: Among the 255 strains of Shigella flexneri preserved in CMCC (B) from 1934 to 1965, 79 were carrying gtrⅡ gene, of which 19 strains and 1 strain were agglutinated with the Y serotype and X serotype, respectively, and furthermore, the multiplex PCR assays results showed serotypes 2a and 2b, respectively, and the strains were considered to have serotype conversion. The 20 strains carrying the gtrⅡ gene showed multiple nucleotide mutations. Besides 3 strains of 3 amino acid mutations, the amino acid sequences of the other 17 strains showed a stop codon in advance, resulting in functional inactivation of gtrⅡ. PFGE analysis revealed that the similarity between the serotype Y strain carrying the gtrⅡ gene and the serotype 2a strain was 75.8%-100%, and the similarity between the serotype X strain carrying the gtrⅡ gene and the serotype 2b strain was 81.6%-100%. Conclusion: Mutations in the gtrⅡ gene are more complicated in serotype-transforming Shigella flexneri serotype Y or X strains. Molecular typing suggests that the serotype-transforming Shigella flexneri serotype Y or X strains may be derived from the Shigella flexneri serotype 2a or 2b, and advance the serotype conversion to 1949.

Z-Score transformation of ADC values: A way to universal cut off between malignant and benign lymph nodes.

PURPOSE: To determine the possibility of a universal cut off value between benign and malignant lymph nodes in patients with tumour by Z-Score transformation method. MATERIALS AND METHODS: Diffusion weighted imaging, ADC measurements of malignant and benign lymph nodes of 6 studies (4 body parts), conducted for 5 times, in two institutions with variable technical details were analyzed in their original value as well as the standardized Z-Score value. The standardized Z-Score value was obtained by subtracting the population mean of the control group from an individual raw score and then dividing the difference by the population standard deviation of the control group. General cut off values were obtained by both Mega-analysis by receiver operator characteristic curve analysis, when data from the 6 studies were combined and Meta-analysis with weighting coefficients and cut off values of the six individual studies. Sensitivity, specificity and accuracy with cut offs from individual studies, meta-analysis and mega-analysis were calculated. Kappa test was performed to assess the consistency of diagnostic test accuracy, between optimized cut offs of individual studies and the proposed universal cut offs obtained from meta-analysis and mega-analysis. RESULTS: The ADC values of benign and malignant lymph nodes are significantly different, but with large overlap across the studies. The overlap can be minimized by Z-Score transformation. The result of ROC analysis of the collective Z-Score transformed ADC values of 6 studies was superior to that of the collective original ADC values (sensitivity: 87.4% versus 67.2%, specificity: 90.5% versus 87.9%, accuracy: 89.6% versus 81.4%). The universal Z-Score cut off from Meta-analysis is also better than the original ADC cut off (sensitivity: 82.8% versus 76.3%, specificity 92.6% versus 62.9%, accuracy 89.6% versus 67.1%). Applied to the individual studies, the universal transformed Z-Score cut offs produced superior consistency with the individual optimal cut offs (individual and meta Z-Score: 0.7228-0.9793; individual and mega Z-Score: 0.7111-0.9169) compared with the universal original ADC cut offs (individual and meta ADC: 0.3030-1.0000; individual and mega ADC 0.3268-0.9618). CONCLUSION: Z-Score transformation could minimize inter-study variations due to heterogeneity of MR systems and sequence parameters, and provide a more consistent universal cut off value between benign and malignant nodes across studies.

MicroRNA-185-5p modulates chemosensitivity of human non-small cell lung cancer to cisplatin via targeting ABCC1.

OBJECTIVE: MicroRNA-185-5p (miR-185-5p) dysregulation is found in various human cancers. Our purpose is to investigate the association of miR-185-5p expression with the sensitivity of non-small cell lung cancer (NSCLC) to cisplatin. MATERIALS AND METHODS: Real-time PCR or Western blot assay was performed to detect the expression of mature miR-185-5p and ATP-binding cassette, subfamily C, member 1 (ABCC1) protein. Cell lines with abnormal expression of miR-185-5p were generated using miR-185-5p inhibitor and mimics. The viabilities of treated cells were analyzed using MTT assay. Cell apoptosis was evaluated by TUNEL assay. Apoptosis-related protein expressions were tested by Western blot. Dual-luciferase assay was applied to assess the target gene of miRNA. RESULTS: The expression level of miR-185-5p in A549 cell line was significantly higher than that in A549/DDP cell line (p < 0.05). Transfection of miR-185-5p mimics increased the sensitivity of A549 cells to cisplatin and the expression of an apoptosis-related factor, and restrained cell proliferation. MiR-185-5p inhibitor promoted cisplatin resistance and cell growth in A549 cells, and declined apoptosis-related factor levels. ABCC1 was verified as the target gene of miR-185-5p. MiR-185-5p exhibited negative correlation with ABCC1 in A549/DDP cells. CONCLUSIONS: The results of the present study demonstrated that inhibition of miR-185-5p was involved in chemo-resistance of NSCLC cells to cisplatin via down-regulating ABCC1.

Predictors of active cancer thromboembolic outcomes: validation of the Khorana score among patients with lung cancer.

UNLABELLED: Essentials Venous thromboembolism (VTE) prevention strategies require effective risk assessment models. We sought to validate the Khorana Risk Score (KRS) in patients with lung cancer. A high KRS was not predictive of VTE but was independently associated with all-cause mortality. Our findings stress the need for a lung cancer-specific VTE risk assessment model. SUMMARY: Objectives Lung cancer is strongly associated with venous thromboembolism (VTE), but primary prevention against VTE is not a validated management strategy. Risk assessment models will be necessary for efficient implementation of preventative strategies. Materials and methods Utilizing a prospectively collected lung cancer database, we aimed to validate the Khorana Risk Score (KRS) in the prediction of VTE among patients with lung cancer. VTE events were retrospectively identified by reviewers unaware of the clinical prediction score calculation. The association between KRS and the risk of VTE was examined using cumulative incidence function with competing risk models. Mortality prediction was evaluated as a secondary outcome. Results We included 719 patients in our review. The patients were predominantly older men with non-small cell lung cancer and 40% had metastatic disease at inception. The median follow-up was 15.2 months. There were 83 VTEs (11.5%) and 568 (78.8%) patients died. A high KRS (cumulative incidence, 12.4%; 95% confidence interval [CI], 6.4-20.5%) was not associated with VTE compared with an intermediate score (cumulative incidence, 12.1%; 95% confidence interval, 9.5-15.0%) in both univariate and multivariable analyses. However, a high KRS was a predictor of mortality (hazard ratio, 1.7; 95% CI, 1.4-2.2). Conclusions Among patients with lung cancer, the KRS did not stratify the patients at the highest risk of VTE. Improved risk stratification methods are needed for this group of patients prior to implementing a primary prevention strategy.

Positive selection sites in tertiary structure of Leguminosae chalcone isomerase 1.

Isoflavonoids and the related synthesis enzyme, chalcone isomerase 1 (CHI1), are unique in the Leguminosae, with diverse biological functions. Among the Leguminosae, the soybean is an important oil, protein crop, and model plant. In this study, we aimed to detect the generation pattern of Leguminosae CHI1. Genome-wide sequence analysis of CHI in 3 Leguminosae and 3 other closely related model plants was performed; the expression levels of soybean chalcone isomerases were also analyzed. By comparing positively selected sites and their protein structures, we retrieved the evolution patterns for Leguminosae CHI1. A total of 28 CHI and 7 FAP3 (CHI4) genes were identified and separated into 4 clades: CHI1, CHI2, CHI3, and FAP3. Soybean genes belonging to the same chalcone isomerase subfamily had similar expression patterns. CHI1, the unique chalcone isomerase subfamily in Leguminosae, showed signs of significant positive selection as well as special expression characteristics, indicating an accelerated evolution throughout its divergence. Eight sites were identified as undergoing positive selection with high confidence. When mapped onto the tertiary structure of CHI1, these 8 sites were observed surrounding the enzyme substrate only; some of them connected to the catalytic core of CHI. Thus, we inferred that the generation of Leguminosae CHI1 is dependent on the positively selected amino acids surrounding its catalytic substrate. In other words, the evolution of CHI1 was driven by specific selection or processing conditions within the substrate.

Arterial embolization of massive hepatocellular carcinoma with lipiodol and gelatin sponge.

BACKGROUND: Transarterial chemoembolization (TACE) has been used to treat unresectable massive hepatocellular carcinoma (HCC). Lots of embolic agents have been applied in embolization because of it can decrease patient discomfort and side-effects. AIM: The aim was to evaluate the clinical efficacy and safety of TACE with lipiodol and gelatin sponge. MATERIALS AND METHODS: A total of 109 patients with massive HCC (the size of tumor >10 cm and unresectable) from January 2011 to August 2014 in our institution was divided into group A and group B based on the different embolitic agents. Before and about 1-month after each case of TACE, clinical and biological data such as tumor size, Child-Pugh stage, serum alpha-fetoprotein (AFP), complications, were recorded at the same time. RESULTS: In group A, the diameter of the tumor reduced from 12.57 ± 1.26 cm to 9.04 ± 0.89 cm. No patient was complete response (CR), partial response (PR) 36, stable disease (SD) 7 and PD 6; in group B, the diameter of tumor decreased from 12.08 ± 1.42 cm to 8.43 ± 1.05 cm, CR 0, but PR 27, SD 18 and PD 15. RR in group A was significantly higher than in group B (P < 0.05).The change of Child-Pugh stage and AFP pre- and post-operative in group A can be found significantly better than in group B. CONCLUSIONS: TACE with lipiodol and gelatin sponge is a highly effective for massive HCC.

Transcatheter arterial embolization of acute gastrointestinal tumor hemorrhage with Onyx.

BACKGROUND: Endovascular embolization has been used to control gastrointestinal tumor bleeding. Lots of embolic agents have been applied in embolization, but liquid embolic materials such as Onyx have been rarely used because of concerns about severe ischemic complications. AIM: To evaluate the clinical efficacy and safety of transcatheter arterial embolization (TAE) with Onyx for acute gastrointestinal tumor hemorrhage. MATERIALS AND METHODS: Between September 2011 and July 2013, nine patients were diagnosed as acute gastrointestinal tumor hemorrhage by clinical feature and imaging examination. The angiographic findings were extravasation of contrast media in the five patients. The site of hemorrhage included upper gastrointestinal bleeding in seven cases and lower gastrointestinal bleeding in two cases. TAE was performed using Onyx in all the patients, and the blood pressure and heart rate were monitored, the angiographic and clinical success rate, recurrent bleeding rate, procedure related complications and clinical outcomes were evaluated after therapy. The clinical parameters and embolization data were studied retrospectively. RESULTS: All the patients (100%) who underwent TAE with Onyx achieved complete hemostasis without rebleeding and the patients were discharged after clinical improvement without a second surgery. No one of the patients expired during the hospital course. All the patients were discharged after clinical improvement without a second surgery. Postembolization bowel ischemia or necrosis was not observed in any of the patients who received TAE with Onyx. CONCLUSIONS: TAE with Onyx is a highly effective and safe treatment modality for acute gastrointestinal tumor hemorrhage, even with pre-existing coagulopathy.

Comparison of patients undergoing laser vaporization of the prostate versus TURP using the ACS-NSQIP database.

BACKGROUND: The introduction of laser therapies for the management of bladder outlet obstruction in men with BPH has challenged the gold standard treatment, TURP. We sought to compare the changing clinical characteristics of patients undergoing TURP and laser vaporization of the prostate (LVP) over time. METHODS: The American College of Surgeons National Surgical Quality Improvement Program (ACS-NSQIP) database was queried for men who underwent TURP and LVP from 2007 to 2012. Patient demographics, clinical and intraoperative characteristics and 30-day postoperative outcomes were analyzed. RESULTS: In all, 12,645 men met inclusion criteria, of whom 65% underwent TURP and 35% underwent LVP. Overall, men undergoing TURP were more likely to be scheduled as an emergency (3% vs. 1%, P<0.001), have shorter operative times (53 vs. 56 min, P<0.001), longer hospital stays (2.4 vs 1.0 days, P<0.001), more frequent blood transfusions (2.1% vs. 0.6%, P<0.001) and more postoperative complications including: pneumonia (0.5% vs. 0.3%, P=0.02), septic shock (0.3% vs. 0.1%, P=0.045), and reoperation within 30 days (2.2% vs. 1.4%, P=0.06). However, between 2007 and 2012, there was a significant trend for men undergoing TURP to have increased functional independence (93-96%, P<0.01) and American Society of Anesthesiology (ASA) Physical Class I categorization (0.6-5.1%, P<0.001). In contrast, over the same time period, there was a trend for men undergoing LVP to be significantly older (71-73 years, P<0.001) and have an increased hospital stay (0.50 days to 1.30 days, P=0.03). CONCLUSIONS: Statistically significant differences in clinical characteristics of patients undergoing TURP and LVP have historically existed. However, since 2007, the characteristics of patients undergoing LVP and TURP have changed significantly. Further studies are required to compare these patient characteristics with specific urologic variables and to evaluate clinically significant changes in these cohorts.

Complication prevalence following use of tutoplast-derived human acellular dermal matrix in prosthetic breast reconstruction: a retrospective review of 203 patients.

Use of human acellular dermal matrix (ADM) during prosthetic breast reconstruction has increased. Several ADM products are available produced by differing manufacturing techniques. It is not known if outcomes vary with different products. This study reports the complication prevalence following use of a tutoplast-derived ADM (T-ADM) in prosthetic breast reconstruction. We performed a retrospective chart review of 203 patients (mean follow-up times 12.2 months) who underwent mastectomy and immediate prosthetic breast reconstruction utilizing T-ADM, recording demographic data, surgical indications and complication (infection, seroma, hematoma, wound healing exceeding three weeks and reconstruction failure). During a four-year period, 348 breast reconstructions were performed Complications occurred in 16.4% of reconstructed breasts. Infection occurred in 6.6% of breast reconstructions (3.7% - major infection, requiring intravenous antibiotics and 2.9% minor infection, requiring oral antibiotics only). Seromas occurred in 3.4% and reconstruction failure occurred in 0.6% of breast reconstructions. Analysis suggested that complication prevalence was significantly higher in patients with a BMI >30 (p = 0.03). The complication profile following T-ADM use is this series is comparable to that reported for with other ADM products. T-ADM appears to be a safe and acceptable option for use in ADM-assisted breast reconstruction.

ConfMatch: automating electron-density map interpretation by matching conformations.

Building a protein model from the initial three-dimensional electron-density distribution (density map) is an important task in X-ray crystallography. This problem is computationally challenging because proteins are extremely flexible. The algorithm ConfMatch is a global real-space fitting procedure in torsion-angle space. It solves this 'map-interpretation' problem by matching a detailed conformation of the molecule to the density map (conformational matching). This 'best-match' structure is defined as one which maximizes the sum of the density at atom positions. ConfMatch is a practical systematic algorithm based on a branch-and-bound search. The most important idea of ConfMatch is an efficient method for computing accurate bounds. ConfMatch relaxes the conformational matching problem, a problem which can only be solved in exponential time, into one which can be solved in polynomial time. The solution to the relaxed problem is a guaranteed upper bound for the conformational matching problem. In most empirical cases, these bounds are accurate enough to prune the search space dramatically, enabling ConfMatch to solve structures with more than 100 free dihedral angles. Experiments have shown that ConfMatch may be able to automate the interpretation of density maps of small proteins.

Knowing and approaching hope as human experience: implications for the medical-surgical nurse.

Hospitalized patients frequently share the mystery of their hopes and dreams. Through a deepened understanding and an alternative approach in practice, nurses can come to know patients' hopes and work with them in ways that enhance their quality of life.

Developing a concept of hope from a human science perspective.

This article explores concept development from a human science perspective and uses concept inventing, a method described by Parse, for developing the concept of hope as inspired by the Taiwanese folk song "Mending a Torn Fish Net." The synthesized definition is hope is resolute picturing of the possibilities amid the restrictions-opportunities of communion-aloneness while creatively metamorphosing. This definition is explicitly connected to the three principles of Parse's theory of human becoming. Reflections on research and practice are provided.

Uncv (uncovered): a new mutation causing hairloss on mouse chromosome 11.

A pair of mutant mice with a first sparse coat appeared spontaneously in the production stock of BALB/c mice with a normal coat. After being sib-mated, they produced three phenotypes in their progeny: mice with normal hair, mice with a first sparse coat and then a fuzzy coat, and uncovered mice. Genetic studies revealed the mutants had inherited an autosomal monogene that was semi-dominant. By using 11 biochemical loci--Idh, Car2, Mup1, Pgm1, Hbb, Es1, Es10, Gdc, Ce2, Mod1 and Es3--as genetic markers, two-point linkage tests were made. The results showed the gene was assigned to chromosome 11. The result of a three-point test with Es3 and D11Mit8 (microsatellite DNA) as markers showed that the mutation was linked to Es3 with the recombination fraction 7.89 +/- 2.19%, and linked to D11Mit8 with the recombination fraction 26.30 +/- 3.57%. The recombination fraction between Es3 and D11Mit8 was 32.90 +/- 3.81%. It is suggested that the mutation is a new genetic locus that affected the skin and hair structure of the mouse. The mutation was named uncovered, with the symbol Uncv. Further studies showed the mutation affected not only the histology of skin and hair but also the growth and reproductive performance of the mice. The molecular characterization of the Uncv locus needs to be further studied.