Aortic root morphometry revisited-Clinical implications for aortic valve interventions.

The complex anatomy of the aortic root is of great importance for many surgical and transcatheter cardiac procedures. Therefore, the aim of this study was to provide a comprehensive morphological description of the nondiseased aortic root. We morphometrically examined 200 autopsied human adult hearts (22.0% females, 47.9 ± 17.7 years). A meticulous macroscopic analysis of aortic root anatomy was performed. The largest cross-section area of the aortic root was observed in coaptation center plane (653.9 ± 196.5 mm2), followed by tubular plane (427.7 ± 168.0 mm2) and basal ring (362.7 ± 159.1 mm2) (p < 0.001). The right coronary sinus was the largest (area: 234.3 ± 85.0 mm2), followed by noncoronary sinus (218.7 ± 74.8 mm2) and left coronary sinus (201.2 ± 78.08 mm2). The noncoronary sinus was the deepest, followed by right and left coronary sinus (16.4 ± 3.2 vs. 15.9 ± 3.1 vs. 14.9 ± 2.9 mm, p < 0.001). In 68.5% of hearts, the coaptation center was located near the aortic geometric center. The left coronary ostium was located 15.6 ± 3.8 mm above sinus bottom (within the sinus in 91.5% and above sinutubular junction in 8.5%), while for right coronary ostium, it was 16.2 ± 3.5 mm above (83.5% within sinus and 16.5% above). In general, males exhibited larger aortic valve dimensions than females. A multiple forward stepwise regression model showed that anthropometric variables might predict the size of coaptation center plane (age, sex, and heart weight; R2 = 31.8%), tubular plane (age and sex; R2 = 25.6%), and basal ring (age and sex; R2 = 16.9%). In conclusion, this study presents a comprehensive analysis of aortic-root morphometry and provides a platform for further research into the intricate interplay between structure and function of the aortic root.

Pulmonary valve morphometry revisited: Clinical implications for valvular and supravalvular interventions.

In this cadaver-based study, we aimed to present a novel approach to pulmonary valve (PV) anatomy, morphometry, and geometry to offer comprehensive information on PV structure. The 182 autopsied human hearts were investigated morphometrically. The largest PV area was seen for the coaptation center plane, followed by basal ring and the tubular plane (626.7 ± 191.7 mm2 vs. 433.9 ± 133.6 mm2 vs. 290.0 ± 110.1 mm2 , p < 0.001). In all leaflets, fenestrations are noted and occur in 12.5% of PVs. Only in 31.3% of PVs, the coaptation center is located in close vicinity of the PV geometric center. Similar-sized sinuses were found in 35.7% of hearts, in the remaining cases, significant heterogeneity was seen in size. The mean sinus depth was: left anterior 15.59 ± 2.91 mm, posterior: 16.04 ± 2.82 mm and right anterior sinus: 16.21 ± 2.81 mm and the mean sinus height: left anterior 15.24 ± 3.10 mm, posterior: 19.12 ± 3.79 mm and right anterior sinus: 18.59 ± 4.03 mm. For males, the mean pulmonary root perimeters and areas were significantly larger than those for females. Multiple forward stepwise regression model showed that anthropometric variables might predict the coaptation center plane (sex, age, and heart weight; R2  = 33.8%), tubular plane (sex, age, and BSA; R2  = 20.5%) and basal ring level area (heart weight and sex; R2  = 17.1%). In conclusion, the largest pulmonary root area is observed at the coaptation center plane, followed by the basal ring and tubular plane. The PV geometric center usually does not overlap valve coaptation center. Significant heterogeneity is observed in the size of sinuses and leaflets within and between valves. Anthropometric variables may be used to predict pulmonary root dimensions.

Radiological Features for Frailty Assessment in Patients Requiring Emergency Laparotomy.

INTRODUCTION: As the number of elderly patients requiring surgical intervention rises, it is believed that frailty syndrome has a greater impact on perioperative course than on chronological age. The aim of this study was to evaluate the efficacy of various imaging features for frailty assessment in patients undergoing emergency laparotomy. METHODS: The study included all patients that qualified for emergency surgery with preoperative CT scans between 2016 and 2020 in the Second Department of General Surgery. Multiple trauma patients were excluded from the analysis. The modified frailty index and brief geriatric assessment were used in the analysis. CT images were reviewed for the assessment of osteopenia, sarcopenia, sarcopenic obesity, renal volume and abdominal aorta calcification rate. RESULTS: A total of 261 patients were included in the analysis. Multivariate logistic regression identified every next ASA class (OR: 4.161, 95%CI: 1.672-10.355, p = 0.002), intraoperative adverse events (OR: 12.397, 95%CI: 2.166-70.969, p = 0.005) and osteopenia (OR: 4.213, 95%CI: 1.235-14.367, p = 0.022) as a risk factor for 30-day mortality. Our study showed that every next ASA class (OR: 1.952, 95%Cl: 1.171-3.256, p = 0.010) and every point of the BGA score (OR: 1.496, 95%Cl: 1.110-2.016, p = 0.008) are risk factors for major complications. CONCLUSIONS: Osteopenia was the best parameter for perioperative mortality risk stratification in patients undergoing emergency surgical intervention. Sarcopenia (measured as psoas muscle area), sarcopenic obesity, aortic calcifications and mean kidney volume do not predict poor outcomes in those patients. None of the radiological markers appeared to be useful for the prediction of perioperative morbidity.

The persistent median artery and its vascular patterns: A meta-analysis of 10,394 subjects.

The presence of a persistent median artery (PMA) has been implicated in the development of compression neuropathies and surgical complications. Due to the large variability in the prevalence of the PMA and its subtypes in the literature, more awareness of its anatomy is needed. The aim of our meta-analysis was to find the pooled prevalence of the antebrachial and palmar persistent median arteries. An extensive search through the major databases was performed to identify all articles and references matching our inclusion criteria. The extracted data included methods of investigation, prevalence of the PMA, anatomical subtype (antebrachial, palmar), side, sex, laterality, and ethnicity. A total of 64 studies (n = 10,394 hands) were included in this meta-analysis. An antebrachial pattern was revealed to be more prevalent than a palmar pattern (34.0% vs. 8.6%). A palmar PMA was reported in 2.6% of patients undergoing surgery for carpal tunnel syndrome when compared to cadaveric studies of adult patients in which the prevalence was 8.6%. Both patterns of PMA are prevalent in a considerable portion of the general population. As the estimated prevalence of the PMA was found to be significantly lower in patients undergoing surgery for carpal tunnel syndrome than those reported in cadaveric studies, its etiological contribution to carpal tunnel syndrome is questionable. Surgeons operating on the forearm and carpal tunnel should understand the anatomy and surgical implications of the PMA and its anatomical patterns.

Microanatomy of the myocardial extensions of the pulmonary valve in light of modern catheter ablation methodology.

INTRODUCTION: The muscular sleeves (or myocardial extensions) derived from the right ventricle infundibulum myocardium are considered the true anatomic substrate for right ventricular outflow tract arrhythmias. METHODS: Pulmonary valve specimens obtained from 65 donors (24.6% females, mean age 45.9 ± 15.8 years) were investigated micro-anatomically. Specimens were histologically processed, stained with Masson's Trichrome, and examined under a light microscope. RESULTS: The myocardial extensions were present in the left anterior pulmonary valve sinus in 86.2% of cases, in the right anterior sinus in 89.2% of cases and in 90.7% of cases in the posterior sinus (p = .699). In 69.2% of examined hearts, the myocardial extensions were present in all sinuses. The mean height of the extensions was 4.12 ± 1.76 (left anterior) versus 3.69 ± 1.47 (right anterior) versus 4.28 ± 1.73 mm (posterior) (p = .137). The myocardial extensions occupied an average of 28.9 ± 10.4% of the left anterior sinus, 26.7 ± 11.2% of the right anterior sinus, and 31.9 ± 11.3% of the posterior sinus (p = .044). Sleeves extending beyond the fibro-arterial transition zone were present in at least one sinus in 33.8% of hearts (in 7.7% (5/65) of the left and right anterior sinuses and 21.5% (14/65) of posterior sinus, p = .021). CONCLUSIONS: The myocardial extensions of the pulmonary valve are common anatomical entities. Although the length of the myocardial sleeves is similar in all pulmonary valve sinuses, their relative extent is greatest in the posterior sinus. Long sleeves that spread beyond the fibro-arterial transition zone were observed in one-third of hearts, predominantly in the posterior sinus. Myocardial and fibrous tissue layer thicknesses varied considerably.

Morphology of the Left Atrial Appendage: Introduction of a New Simplified Shape-Based Classification System.

BACKGROUND: The left atrial appendage (LAA) is a heart structure with known prothrombogenic and pro-arrhythmogenic properties. AIM: The aim of this study was to evaluate the specific anatomy of the LAA and to create a simple classification system based on the shape of its body. METHOD AND RESULTS: This study investigated 200 randomly selected autopsied human hearts (25.0% females, 46.6±19.1 years old). Three (3) types of LAAs were distinguished: the cauliflower type (no bend, limited overall length, compact structure [36.5%]); the chicken wing type (substantial bend in the dominant lobe [37.5%]), and the arrowhead type (no bend, one dominant lobe of substantial length [26.0%]). Additional accessory lobes were present in 55.5% of all LAAs. Significant variations between category types were noted in LAA length (chicken wing: 35.7±9.8 mm, arrowhead: 30.8±10.1 mm, cauliflower: 22.3±9.6 mm [p<0.001]) and in the thickness of pectinate muscles located within the LAA apex (arrowhead: 1.2±0.7 mm; cauliflower: 1.1±0.6 mm; chicken wing: 0.9±0.6 mm [p<0.001]). Left atrial appendage volume and orifice size were not affected by the type of LAA shape. The age of the donor was positively correlated with LAA volume (r=0.29, p=0.005), body length (r=0.26, p=0.012), and area of the orifice (r=0.36, p<0.001). Donors with an oval LAA orifice were significantly older than those with round orifices (50.2±16.6 vs 43.7±20.4 years [p=0.014]) and had significantly heavier hearts (458.2±104.8 vs 409.6±114.1g [p=0.002]). CONCLUSIONS: This study delivered a new simple classification system of the LAA based on its body shape. An increase in age and heart weight was associated with LAA enlargement and a more oval-shaped orifice. Results of current study may help to estimate the different thrombogenic properties associated with each LAA type and be an assistance during planning and performing interventions on LAA.

Identification of CD34+/PGDFRα+ Valve Interstitial Cells (VICs) in Human Aortic Valves: Association of Their Abundance, Morphology and Spatial Organization with Early Calcific Remodeling.

Aortic valve interstitial cells (VICs) constitute a heterogeneous population involved in the maintenance of unique valvular architecture, ensuring proper hemodynamic function but also engaged in valve degeneration. Recently, cells similar to telocytes/interstitial Cajal-like cells described in various organs were found in heart valves. The aim of this study was to examine the density, distribution, and spatial organization of a VIC subset co-expressing CD34 and PDGFRα in normal aortic valves and to investigate if these cells are associated with the occurrence of early signs of valve calcific remodeling. We examined 28 human aortic valves obtained upon autopsy. General valve morphology and the early signs of degeneration were assessed histochemically. The studied VICs were identified by immunofluorescence (CD34, PDGFRα, vimentin), and their number in standardized parts and layers of the valves was evaluated. In order to show the complex three-dimensional structure of CD34+/PDGFRα+ VICs, whole-mount specimens were imaged by confocal microscopy, and subsequently rendered using the Imaris (Bitplane AG, Zürich, Switzerland) software. CD34+/PDGFRα+ VICs were found in all examined valves, showing significant differences in the number, distribution within valve tissue, spatial organization, and morphology (spherical/oval without projections; numerous short projections; long, branching, occasionally moniliform projections). Such a complex morphology was associated with the younger age of the subjects, and these VICs were more frequent in the spongiosa layer of the valve. Both the number and percentage of CD34+/PDGFRα+ VICs were inversely correlated with the age of the subjects. Valves with histochemical signs of early calcification contained a lower number of CD34+/PDGFRα+ cells. They were less numerous in proximal parts of the cusps, i.e., areas prone to calcification. The results suggest that normal aortic valves contain a subpopulation of CD34+/PDGFRα+ VICs, which might be involved in the maintenance of local microenvironment resisting to pathologic remodeling. Their reduced number in older age could limit the self-regenerative properties of the valve stroma.

The interaction of laser radiation with tissue in the aspect of generating the process of decellularization in the preparation of animal origin autologous tissue.

PURPOSE: The aim of the work was to create an appropriate substrate for organ transplantation using bioactive tissue-based scaffold populated by cells of the graft recipient. The purpose of the modeling was to investigate the mechanical effects of wave loading of aortic and pulmonary tissue material. METHODS: The biological properties of tissues of aortic and pulmonary valves were modified by the process of decellularization. The host cells were removed by various physical methods with focus on minimal degradation of the extracellular matrix. Thus, the decellularization process was controlled by histological methods. The tissue decellularization process was simulated by finite element modelling. RESULTS: The mechanical results represented by a displacement at the center of the sample were coherent and the heterogeneity of the distribution of the caves on the surface of the samples was confirmed, both by experiment and in the simulation by the alternate occurrence of local minima and maxima. The latter results from the uneven removal of cells from the effect of the wave causing decellularization were also predicted by the numerical model. Laser radiation had a destructive effect on the components of the extracellular matrix (e.g., collagen and elastic fibers), mainly depending on the fluence and number of pulses in a single exposure. CONCLUSIONS: The differences between the valve tissue materials were shown, and the impact of the process of decellularization on the properties of the tissues was analyzed. It should be emphasized that due to low absorption and high scattering, laser radiation can deeply penetrate the tissue, which allows for effective decellularization process in the entire volume of irradiated tissue.

Anatomy of the left atrial ridge (coumadin ridge) and possible clinical implications for cardiovascular imaging and invasive procedures.

BACKGROUND: The left atrial ridge is a structure located in the left atrium between the left-sided pulmonary veins ostia and the orifice of the left atrial appendage. Since it was commonly misdiagnosed as a thrombus, the ridge is also known as the "coumadin" or "warfarin" ridge. The left atrial ridge is a potential source of arrhythmias and can be an obstacle in ablation procedures. This study aimed to provide information about the occurrence and spatial morphometric characteristics of the left atrial ridge. METHODS AND RESULTS: The macroscopic morphology of the left atrial ridge was assessed in 200 autopsied human hearts. The ridge was observed in 59.5% of specimens and was absent in the remaining 40.5% of cases. The mean length of the ridge was 22.4 ± 5.1 mm. It was wider at its inferior sector when compared to its superior sector (9.1 ± 5.0 vs 7.9 ± 3.2 mm; P = .028). The total wall thickness measured at the cross section of the ridge was significantly larger in the inferior than in superior sector (6.2 ± 3.5 vs 4.3 ± 1.8 mm; P < .001), although the myocardial thickness was significantly larger at the superior sector (3.1 ± 1.4 vs 1.9 ± 0.9 mm in inferior sector, P < .001). CONCLUSION: The left atrial ridge is a variable structure, present in 59.5% of humans. The ridge is significantly wider and thicker at its inferior sector, although the actual myocardial layer present within the ridge is thinner at this location. Knowledge about the left atrial ridge morphology is key in avoiding unnecessary interventions or complications during invasive procedures.

Quadricuspid pulmonary valve: macroscopic and microscopic morphometric examination.

A quadricuspid pulmonary valve obtained upon autopsy of a 26-year-old male was examined. The macroscopic evaluation revealed three normal (posterior, right anterior and left anterior) leaflets and one additional leaflet of the pulmonary valve. Except that, the heart showed neither other anatomical variabilities nor any signs of heart disease. The additional leaflet was located between the left anterior and right anterior leaflets and was significantly smaller in size. Under the microscope, all leaflets showed preservation of the typical, layered structure. The thickness and extracellular matrix composition of the particular layers differed between the leaflets. Right ventricular myocardium (myocardial sleeves) exceeded the level of the hinge line in all three normal leaflets, which was not observed in the additional leaflet. Autonomic nerves and ganglia were not seen in the perivalvular epicardial adipose tissue surrounding the additional leaflet. The sinus wall of all the leaflets revealed typical organization of collagen bundles as well as elastic fibers and showed no signs of disruption. The abnormality seen in the structure of the pulmonary valve is likely to be a result of disturbed embryonic development and may affect the clinical management of patients with such variation.

The median nerve penetrating through the radial head of the flexor digitorum superficialis muscle: Case report.

The forearm is a body region of numerous anatomical variations. Due to its favorable anatomy flexor digitorum superficialis muscle (FDS) is commonly used in tendon transfer surgeries. In this study a unique combination of abnormalities was found in a single forearm: the flexor digitorum superficialis muscle penetrated by the median nerve, one of the flexor digitorum superficialis tendons early division and absence of the palmaris longus muscle. Described variation potentially may lead to the clinical manifestation of the median nerve compression and should be also considered during FDS surgery.

Unusual branching pattern of celiac trunk associated with supernumerary hepatic arteries and abnormal adrenal venous drainage - case study and review of the literature.

Celiac artery (trunk) is one of the three major arteries which arise from abdominal aorta. It's variations not seem to be very uncommon. A routine dissection of a male cadaver at Department of Anatomy Jagiellonian University revealed unusual branching pattern of the celiac trunk with numerous supernumerary hepatic arteries. Additionally unusual venous drainage of the adrenal glands was found. A review of current literature has shown that a changed branching pattern may be important from clinical point of view, with special respect to endovascular procedures, laparoscopic surgery or radiology.

Angiogenesis and pro-angiogenic factors in uterine fibroids - facts and myths.

Uterine leiomyomata present major problem for females. Although they are benign tumors their frequency is associated with many symptoms like infertility, abdominal pain, menorrhagia. Authors based on their own morphological studies and review of the literature try to indicate main factors causing angiogenesis within leiomyomata and its influence on tumor growth. The strongest proangiogenic factor seems to be hypoxia, which stimulates up- and down-regulation of numerous genetically determined substances. Also mechanical pressure acting upon newly growing vessels is one of the factors which may determine formation of so called "vascular pseudocapsule" around the lesion.

Chemical genomic screening of a Saccharomyces cerevisiae genomewide mutant collection reveals genes required for defense against four antimicrobial peptides derived from proteins found in human saliva.

To compare the effects of four antimicrobial peptides (MUC7 12-mer, histatin 12-mer, cathelicidin KR20, and a peptide containing lactoferricin amino acids 1 to 11) on the yeast Saccharomyces cerevisiae, we employed a genomewide fitness screen of combined collections of mutants with homozygous deletions of nonessential genes and heterozygous deletions of essential genes. When an arbitrary fitness score cutoffs of 1 (indicating a fitness defect, or hypersensitivity) and -1 (indicating a fitness gain, or resistance) was used, 425 of the 5,902 mutants tested exhibited altered fitness when treated with at least one peptide. Functional analysis of the 425 strains revealed enrichment among the identified deletions in gene groups associated with the Gene Ontology (GO) terms "ribosomal subunit," "ribosome biogenesis," "protein glycosylation," "vacuolar transport," "Golgi vesicle transport," "negative regulation of transcription," and others. Fitness profiles of all four tested peptides were highly similar, particularly among mutant strains exhibiting the greatest fitness defects. The latter group included deletions in several genes involved in induction of the RIM101 signaling pathway, including several components of the ESCRT sorting machinery. The RIM101 signaling regulates response of yeasts to alkaline and neutral pH and high salts, and our data indicate that this pathway also plays a prominent role in regulating protective measures against all four tested peptides. In summary, the results of the chemical genomic screens of S. cerevisiae mutant collection suggest that the four antimicrobial peptides, despite their differences in structure and physical properties, share many interactions with S. cerevisiae cells and consequently a high degree of similarity between their modes of action.

Humanized mouse model used to monitor MUC gene expression in nasal polyps and to preclinically evaluate the efficacy of montelukast in reducing mucus production.

OBJECTIVES: To determine whether MUC gene expression could be down-regulated in nasal polyps by the leukotriene receptor antagonist montelukast, we developed a system in which nondisrupted human nasal polyps could be successfully implanted into severely immunocompromised mice, and in which the histopathology of the original nasal polyp tissue could be preserved for long periods. In addition, the histopathologic changes in the human nasal polyps were carefully examined to determine the origin of the submucosal glands (SMGs) that develop in true nasal polyps found in the anterior third of the nose. METHODS: Small, nondisrupted pieces of human nasal polyp tissues were subcutaneously implanted into NOD-scid IL-2rgamma(null) mice. Xenograft-bearing mice were treated with either montelukast or saline solution. Xenografts at 8 to 12 weeks after implantation were examined histologically, and expression of MUC genes 4, 5AC, and 7 was studied in the polyps before implantation and in the 8-week xenograft. Alzet pumps were inserted into the mice, and montelukast (Singulair) was continuously delivered to determine its effect on goblet cell hyperplasia, mucus production, and the enlargement of nasal polyps over an 8-week period. RESULTS: The xenografts were maintained in a viable and functional state for up to 3 months and retained a histopathology similar to that of the original tissue, but with a noticeable increase in goblet cell hyperplasia and marked mucus accumulation in the SMGs. MUC4 and MUC5AC were significantly increased in the xenograft 8 weeks after implantation, but MUC7 was significantly decreased compared to the preimplantation polyps. Inasmuch as MUC7 is found exclusively in serous glands, the findings suggest that serous glands are not found in polyps in the anterior third of the nose. The histopathologic findings confirm the original findings of Tos et al suggesting that the SMGs are derived from pinching-off of the epithelium of the enlarging polyp following inflammatory changes. These SMGs have the same epithelium as surface epithelium and consist of multiple goblet cells that secrete periodic acid Schiff stain-positive mucin into the interior of the SMGs. A progressive increase in the volume of the xenografts was observed, with little or no evidence of mouse cell infiltration into the human leukocyte antigen-positive human tissue. An average twofold increase in polyp volume was found 2 months after engraftment. Montelukast did not decrease the growth of the xenograft in the 8-week NOD-scid mice, nor did it affect MUC gene expression. CONCLUSIONS: The use of innate and adaptive immunodeficient NOD-scid mice homozygous for targeted mutations in the IL-2 gamma-chain locus NOD-scid IL-2r gamma(null) for establishing engraftment of nondisrupted pieces of human nasal polyp tissues represents a significant advancement in studying chronic inflammation over a long period of time. In the present study, we utilized this humanized mouse model to confirm our prediction that MUC genes 4 and 5AC are highly expressed and significantly increased over those of preimplanted polyps. The overexpression of these 2 MUC genes correlates with both the goblet cell hyperplasia and the excessive mucus production that are found in nasal polyp xenografts. MUC7, which is primarily associated with the submucosa, as opposed to MUC4 and MUC5AC, which are primarily expressed in the epithelium, was significantly decreased in the nasal polyp xenografts. Montelukast had no significant effect on MUC gene expression in the xenografts. In addition to the MUC gene expression patterns, the histology of the xenografts supports the concept that mucinous glands that are characteristic of true nasal polyps are significantly different from those in the mucosa found in the lateral wall of the nose in patients with chronic sinusitis without nasal polyps. The mucinous glands seen in nasal polyps (which appear to be derived from an invagination of hyperplastic epithelial mucosa containing large numbers of goblet cells) are histologically distinct from the seromucinous glands found in the submucosa of hyperplastic middle turbinates. The data presented here establish a humanized mouse model as a viable approach to study nasal polyp growth, to assess the therapeutic efficacy of various drugs in this chronic inflammatory disease, and to contribute to our understanding of the pathogenesis of this disease.

Antimicrobial peptide MUC7 12-mer activates the calcium/calcineurin pathway in Candida albicans.

MUC7 12-mer is a cationic antimicrobial peptide derived from the N-terminal region of human low-molecular-weight salivary mucin. In order to gain new insights into the modes of action of the 12-mer against opportunistic fungal pathogen Candida albicans, we examined changes in the gene expression profile of C. albicans upon exposure to this peptide. Cells at an early logarithmic phase were exposed to 6 muM peptide and grown until an OD(600 nm) of approximately 0.4 was reached. Changes in gene expression were determined by microarray analysis and showed that 19 out of the total of 531 genes, whose expression was elevated in response to the peptide, are regulated by the calcium/calcineurin signalling pathway. Inactivation of this pathway by deletions, or by FK506, caused hypersensitivity to the peptide, demonstrating the importance of this pathway to the defense of C. albicans against the MUC7 peptide. Other differentially expressed genes that were detected include those encoding subunits of proteasome, and genes involved in cell stress, iron metabolism, cell wall maintenance and small-molecule transport. The presented results suggest that the calcium/calcineurin signalling pathway plays a role in the adaptation of C. albicans to the MUC7 antimicrobial peptide.

Exploring the mode of action of antimicrobial peptide MUC7 12-mer by fitness profiling of Saccharomyces cerevisiae genomewide mutant collection.

The MUC7 12-mer (RKSYKCLHKRCR) is a cationic antimicrobial peptide derived from the human salivary mucin MUC7. To study its effect/mechanism of action on fungi, we performed a fitness screen of a tagged, diploid, homozygous gene deletion mutant pool of the yeast Saccharomyces cerevisiae grown in the presence of the MUC7 peptide. Forty-five strains exhibiting reduced fitness and 13 strains exhibiting increased fitness (sensitivity or resistance, respectively) were identified by hybridization intensities to tag arrays. The strongest fitness defects were observed with deletions in genes encoding elements of the RIM101 signaling pathway (regulating response to alkaline and neutral pH and other environmental conditions) and of the endosomal sorting complex required for transport (ESCRT; functioning mainly in protein sorting for degradation, but also required for activation of the RIM101 pathway). Other deletions identified as conferring fitness defect or gain are in genes associated with a variety of functions, including transcription regulation, protein trafficking, transport, metabolism, and others. The results of the pool fitness screen were validated by a set of mutant strains tested individually in the presence of the MUC7 12-mer. All tested RIM101-related deletion strains showing fitness defects confirmed their sensitivities. Taken together, the results led us to conclude that deletions of genes associated with the RIM101 pathway confer sensitivity to the peptide by preventing activation of this pathway and that this stress response plays a major role in the protection of S. cerevisiae against damage inflicted by the MUC7 12-mer peptide.

Proteomic and metabolic characterization of a Candida albicans mutant resistant to the antimicrobial peptide MUC7 12-mer.

MUC7 12-mer is a cationic peptide derived from the N-terminal portion of human mucin MUC7, exhibiting potent antibacterial and antifungal properties. To advance our knowledge regarding the mechanisms of action of MUC7 peptide against an opportunistic fungal pathogen Candida albicans, we sought to develop and characterize mutant(s) resistant to this peptide. One of the selected mutants, designated #37, was much less susceptible to the MUC7 12-mer in a killing assay than the parental strain (ED(50)>40 vs. c. 6 microM, respectively). Difference gel electrophoresis (DIGE) analysis of the mutant revealed elevation of several glycolytic enzymes. The mutant also exhibited lowered ATP contents along with a relatively lower rate of oxygen consumption, as well as inability to grow on nonfermentable carbon sources. These properties are likely to be associated with changes in metabolic regulation, rather than lack of functional mitochondria, as determined by rhodamine 123 staining. Analysis of interaction between fluorescently labeled peptide and cells of both strains revealed that resistance of the mutant #37 is associated with changes in the process of transition between surface-bound state of the peptide to its internalization marking cell death.

Persistent regional downregulation in mitochondrial enzymes and upregulation of stress proteins in swine with chronic hibernating myocardium.

Hibernating myocardium is accompanied by a downregulation in energy utilization that prevents the immediate development of ischemia during stress at the expense of an attenuated level of regional contractile function. We used a discovery based proteomic approach to identify novel regional molecular adaptations responsible for this phenomenon in subendocardial samples from swine instrumented with a chronic LAD stenosis. After 3 months (n=8), hibernating myocardium was present as reflected by reduced resting LAD flow (0.75+/-0.14 versus 1.19+/-0.14 mL x min(-1) x g(-1) in remote) and wall thickening (1.93+/-0.46 mm versus 5.46+/-0.41 mm in remote, P<0.05). Regionally altered proteins were quantified with 2D Differential-in-Gel Electrophoresis (2D-DIGE) using normal myocardium as a reference with identification of candidates using MALDI-TOF mass spectrometry. Hibernating myocardium developed a significant downregulation of many mitochondrial proteins and an upregulation of stress proteins. Of particular note, the major entry points to oxidative metabolism (eg, pyruvate dehydrogenase complex and Acyl-CoA dehydrogenase) and enzymes involved in electron transport (eg, complexes I, III, and V) were reduced (P<0.05). Multiple subunits within an enzyme complex frequently showed a concordant downregulation in abundance leading to an amplification of their cumulative effects on activity (eg, "total" LAD PDC activity was 21.9+/-3.1 versus 42.8+/-1.9 mU, P<0.05). After 5-months (n=10), changes in mitochondrial and stress proteins persisted whereas cytoskeletal proteins (eg, desmin and vimentin) normalized. These data indicate that the proteomic phenotype of hibernating myocardium is dynamic and has similarities to global changes in energy substrate metabolism and function in the advanced failing heart. These proteomic changes may limit oxidative injury and apoptosis and impact functional recovery after revascularization.