Rhinophyma: review and update

Learning Objectives: After studying this article, the participant should be able to discuss: 1. Clinical features and anatomy of rhinophyma. 2. The etiology and epidemiology of rhinophyma. 3. Associated diagnosis that can complicate rhinophyma. 4. Common nonsurgical and surgical therapies for rhinophyma. 5. A safe and integrated treatment plan for the patient with rhinophyma.     

Rhinophyma: dispelling the myths

Rhinophyma is a relatively common condition in the west of Scotland. The Canniesburn Plastic Surgery Unit receives 12 to 13 new patients per year for surgical treatment. The reported incidence of simultaneous carcinoma in the setting of rhinophyma is on the order of 15 to 30 percent. There are conflicting reports about the association between alcohol and rhinophyma in the literature, and these are supported with little or no statistical evidence. Retrospective epidemiologic data on 45 cases of rhinophyma are presented. An audit of case notes was performed to examine histology and also alcohol consumption in these cases. The authors found no coincidental malignancies at the time of surgery, which is contrary to many previous publications. The alcohol consumption of the rhinophyma cases was compared with that of a control group that consisted of 48 men presenting for blepharoplasty. The series did not demonstrate a positive association between alcohol and rhinophyma when compared with a similar cohort of patients presenting for blepharoplasty surgery (p > 0.20) or with statistics available from the Scottish Health Survey.     

Antimicrobial effect of silver-impregnated cellulose: potential for antimicrobial therapy

Background  

Silver has long been known to have antimicrobial activity. To incorporate this property into multiple applications, a silver-impregnated cellulose (SIC) with low cytotoxicity to human cells was developed. SIC differs from other silver treatment methods in that the leaching of silver particles is non-existent and the release of ionic silver is highly controlled.     

Effect of electrocautery of nonovulated day 1 follicles on subsequent morphological variation among day 11 porcine embryos

One hundred and thirteen crossbred gilts were used in three experiments to examine the relationship between the pattern or sequence of ovulation and subsequent variation in the morphology of Day 11 embryos. In the first experiment, the percentage of follicles that had ovulated was determined in individual gilts at 26, 30, 34, or 38 h after the onset of estrus (n = 20) and 39, 41, 43, 45, or 47 h post-injection of human chorionic gonadotropin (n = 25; hCG, 1000 IU). The second experiment consisted of observing the percentage of follicles ovulated in 52 additional gilts at 34 h after the onset of estrus (Day 0). In the third experiment, the morphological variation among littermate embryos was compared on Day 11 between sham-operated control gilts (n = 8) and gilts whose nonovulated follicles were destroyed by electrocautery (n = 8) on Day 1. Results of these experiments indicated that the pattern of ovulation in gilts was skewed (p less than 0.01). Ovulation, induced with hCG, appeared to occur in a majority of follicles during a short period of time, whereas the remaining ovulations occurred over a longer interval. Of the 57 gilts observed at 34 h after natural estrus, ovaries of 25 gilts contained corpora hemorrhagica (CH) and follicles; one gilt had 1 CH and 17 follicles, and 24 others had 10-17 CH with 1-4 follicles remaining. Destruction of these nonovulated follicles resulted in a more (p less than 0.01) uniform group of Day 11 embryos and with fewer (p less than 0.05) small embryos. These data demonstrated that the pattern of ovulation may affect morphological variation in embryonic development such that some of the later ovulating follicles may represent smaller embryos within a litter.     

Experimental study on the treatment of intracerebral glioma xenograft with human cytokine-induced killer cells

Objective. To investigate the phenotype changes and proliferation activities of cytokine-induced killer cells (CIKs) and lymphokine-activated killer cells (LAKs) from healthy donor, and the cytotoxicities of CIKs and LAKs to human in vitro glioma cell lines U251 and U87. Therapy of CIK intratumoral injection was evaluated in nude mouse models. Methods. CIK cells were induced from peripheral blood mononuclear cells (PBMC) of healthy donors with multiple cytokines. Phenotype analysis of CIKs and LAKs was performed with flow cytometer (FCM). The specific cytotoxicities of CIKs and LAKs against cell line U251 and U87 were determined by LDH method. After intracerebral injection of CIKs, the distribution of CIKs and the inflammatory reaction of their surrounding brain tissue were observed through continuous pathological sections. In vivo anti-tumor activity of CIKs was evaluated in athymic nude mice with intracerebral xenotransplanted U251 glioma by MRI. Results. Amount of CIKs was increased (49.83+/-2.04) times and double positive cells, CD3(+)/CD56(+) cells, were increased from (3.36+/-1.85%) to (44.07+/-14.14%) with elevated absolute amount over 1000 times after 2 week culture. In vitro experiments demonstrated that compared with LAK, CIKs possessed more obvious cytotoxic activity to U251 and U87. In vivo experiments showed that there was no severe inflammatory reaction in brain tissue. CIKs can markedly inhibit intracranial xenotransplanted glioma growth by intracranial injection (P<0.01). Conclusion. CIKs are a kind of highly effective immune cells which have a strong suppressive effect on growth for in vitro and in vivo glioma. Local injection of CIKs does not produce severe damage to normal brain tissue and is likely to be used in clinical adoptive immunotherapy of intracerebral glioma.     

Analysis of the bending behaviour of porcine xenograft leaflets and of neutral aortic valve material: bending stiffness, neutral axis and shear measurements

Flexibility of the materials used in the construction of bioprosthetic heart valves is essential for proper valve operation. We therefore examined the bending behaviour of glutaraldehyde treated porcine aortic valve cusps in comparison with fresh aortic valve tissue. We repeatedly bent a total of 35 strips of fresh and treated tissue to curvatures ranging from 0.2 to 2.2 mm-1. We compared the stiffness of the two materials between circumferential and radial bending, natural and reverse curvatures and constant or variable tensile stress (0.8-40 kPa). Our results showed a weak positive relationship between bending stiffness and applied tensile stress and a strong positive dependance of stiffness on tissue thickness (t). For the fresh tissue, the bending stiffness increased in proportion to t1.14 while for the glutaraldehyde treated tissue it increased with t2.18. Fourteen strips of fresh and treated tissue were also histologically processed, sectioned and examined with polarized light microscopy. Collagen fiber wavelengths and shear deformations were measured utilizing the tissue banding patterns produced by polarized light microscopy. The neutral axis of bending was found to lie very close to the outer surface of the tissue, suggesting that aortic leaflets have a very low compressive elastic modulus. The shear strains measured in fresh tissue were 10 +/- 2.7% vs 3 +/- 4.4% for the treated, indicating a stiffening of the tissue following glutaraldehyde fixation. We conclude that both natural and bioprosthetic valve cusps have a complex flexural behaviour that cannot be modeled using simple bending principles, although the bioprosthetic material more closely approximates the simple beam than does the fresh. The non-linear elastic modulus, high compressibility and shearing between fiber layers are likely responsible for the observed behaviour of the fresh tissue, while the cross-linking and dehydrating effects of glutaraldehyde are believed to be responsible for the alteration in bending properties observed in the treated tissue. Our study suggests that bioprosthetic valve material does not adequately mimic the mechanics of the natural valve tissue, and that the current glutaraldehyde fixation process eliminates many of the beneficial, stress-reducing properties of the aortic leaflet.     

Isolation of transferrin from porcine gastric mucosa: comparison with porcine serum transferrin

1. An iron-binding glycoprotein has been purified to homogeneity from porcine gastric mucosa. 2. The molecular weight (80,000), amino acid composition, carbohydrate content, N-terminal amino acid sequence, tryptic map, stoichiometry of iron binding (2 mol/mol), visible absorption spectrum of the ferric complex and chromatographic behaviour of the gastric protein are all strikingly similar to the corresponding properties of porcine serum transferrin. 3. The quantity of the gastric protein (1.3 mg/g wet weight) present in the gastric mucosa suggests that it is not serum transferrin (plasma concentration 1.8 mg/ml) contaminating the tissue. 4. A role for transferrin in the uptake of dietary iron by the gastrointestinal tract is proposed.     

Chemotherapeutic treatment of xenograft Spirocerca lupi-associated sarcoma in a murine model

To date, data are not available concerning the effectiveness of chemotherapy in the treatment of Spirocerca lupi-associated esophageal sarcomas. In the present study, we compared the effectiveness of 4 chemotherapeutic agents against S. lupi-associated osteosarcoma, using a xenograft murine model created in our lab. Samples of xenografted osteosarcoma were inoculated subcutaneously into 5 groups (n = 10 each) of 6-wk-old male and female NOD/SCID mice. Tumor-bearing mice were divided into treatment and control groups. The treatment groups were injected with either pegylated liposomal doxorubicin (6 mg/kg, intravenously, n = 9), doxorubicin (6 mg/kg, intravenously, n = 8), carboplatin (60 mg/kg, intraperitoneally, repeated twice at 1-wk intervals for a total of 2 doses, n = 9), or cisplatin (6 mg/kg, intraperitoneally, n = 8). The control group was injected with buffered saline (n = 9). Tumor size was determined by caliper measurements. Compared with the control group, the pegylated liposomal doxorubicin- and doxorubicin-treated groups, but not the carboplatin or cisplatin groups, showed significant inhibition of tumor growth. Our results indicate that doxorubicin-based drugs are effective against S. lupi-associated sarcomas in a mouse xenograft model. Because it is less toxic than doxorubicin, pegylated liposomal doxorubicin is likely the drug of choice for treatment of S. lupi-associated sarcomas. We suggest that combination of doxorubicin or its pegylated form with surgical excision will improve the prognosis of dogs with this disease.     

KBSH parvovirus: comparison with porcine parvovirus.

We compared the molecular, antigenic, and pathogenic properties of KBSH parvovirus to those of porcine parvovirus (PPV) isolate NADL-8. KBSH, propagated in swine testes cells in culture, possessed two major capsid polypeptides of 83 and 64 kilodaltons that were similar in size to those of PPV. KBSH-infected cells also contained an 86-kilodalton nonstructural polypeptide that was identical in size to the PPV nonstructural polypeptide (NS-1). The KBSH polypeptides were structurally similar but not identical to the corresponding PPV polypeptides, as revealed by partial proteolysis mapping. Viral replicative-form DNA from KBSH-infected cells was similar in size to PPV replicative-form DNA and exhibited similar but not identical restriction endonuclease cleavage patterns to that of PPV replicative-form DNA. Antigenically, the two viruses were also very closely related. By using heterologous and homologous antisera, the two viruses were indistinguishable in hemagglutination inhibition and immunoprecipitation assays. However, pathogenically these viruses were dramatically different. NADL-8 caused fetal death when injected into swine fetuses in utero and viremia and high persisting antibody titers when administered orally to weaning-age swine. KBSH-inoculated fetuses were normal in appearance, and pigs orally exposed to KBSH failed to establish viremia and demonstrated only transient antibody titers. Thus, KBSH appears to be a PPV that is very closely related to a highly pathogenic PPV isolate, yet is itself nonpathogenic in swine. This reduced pathogenic potential of KBSH may be attributable to its poor ability to replicate in swine.     

Interaction of N-methyl-anthraniloyl-labelled porcine apolipoprotein A-I with porcine lecithin: Cholesterol acyltransferase: An energy transfer study

• 1.1. To investigate whether a direct protein-protein interaction between apoA-I and lecithin: cholesterol acyltransferase (LCAT) is necessary for the activation of the enzyme, apoA-I was labelled with N-methylisatoic anhydride at lysine residues. The intermolecular resonance energy transfer from tryptophan residues of LCAT (donor) to N-methyl-anthraniloyl (NMA)-labelled apoA-I (NMA-apoA-I) (acceptor) was used as a sensitive fluorescence method for studying molecular interactions.
• 2.2. In the absence of lipids no fluorescence energy transfer was measurable.
• 3.3. Fluorescence energy transfer occurred from LCAT to NMA-apoA-I in the presence of liposomes with phospholipid/cholesterol ratios ranging from 5:1 to 18:1 and regardless whether only 1 or up to 5 NMA-apoA-I molecules resided at the liposome surface.
• 4.4. This indicates a preferred binding of the enzyme directly to or in spatial proximity to the activator protein NMA-apoA-I even if enough space at the liposome surface is available to allow LCAT binding at a distance, where no energy transfer is measurable.

     

Aromatase inhibitors and xenograft studies

Aromatase inhibitors (AIs) have become the front-line choice for treatment of ER+ breast cancer. Nevertheless, although patients are responsive initially, they may acquire resistance and become unresponsive to further treatment. In addition, approximately 25% of breast cancers do not express the estrogen receptor (ERα) and consequently, are innately resistant to endocrine therapy. We have investigated the mechanisms associated with this lack of treatment response using xenograft models. We found that in cells and tumors that acquired resistance to the AI letrozole therapy, expression of the ER was reduced whereas growth factor signally was enhanced, including a marked increase in HER2 expression. Treatment with trastuzumab (HER2 antibody) resulted in a significant down-regulation of HER2 and p-MAPK as well as restoration of ERα expression. Thus, when trastuzumab was added to letrozole treatment at the time of tumor progression, there was significantly prolonged tumor suppression compared to trastuzumab or letrozole alone. This suggests that inhibition of both HER2 and ERα signaling pathways are required for overcoming resistance and restoring treatment sensitivity. ER negative tumors are innately resistant to endocrine therapy. Repression of the ERα has been found to be due to epigenetic modifications such as increased methylation and histone deacetylation. We found that entinostat (ENT), a histone deacetylase inhibitor (HDACi), activated not only expression of ERα but also aromatase in MDA-MB-231 ER-negative breast cancer cells, resulting in their ability to respond to estrogen and letrozole. Treatment with ENT in combination with letrozole significantly reduced tumor growth rate in xenografts compared to control tumors (p < 0.001). ENT plus letrozole treatment also prevented the colonization and growth of MDA-MB-231 cells in the lung with a significant reduction (p < 0.03) in both visible and microscopic foci. These results provide a strong indication for possible use of AIs in combination with HDAC inhibitors for the treatment of ER-negative breast cancer.     

Near-infrared fluorescence imaging of mammalian cells and xenograft tumors with SNAP-tag

Fluorescence in the near-infrared (NIR) spectral region is suitable for in vivo imaging due to its reduced background and high penetration capability compared to visible fluorescence. SNAP(f) is a fast-labeling variant of SNAP-tag that reacts with a fluorescent dye-conjugated benzylguanine (BG) substrate, leading to covalent attachment of the fluorescent dye to the SNAP(f). This property makes SNAP(f) a valuable tool for fluorescence imaging. The NIR fluorescent substrate BG-800, a conjugate between BG and IRDye 800CW, was synthesized and characterized in this study. HEK293, MDA-MB-231 and SK-OV-3 cells stably expressing SNAP(f)-Beta-2 adrenergic receptor (SNAP(f)-ADRβ2) fusion protein were created. The ADRβ2 portion of the protein directs the localization of the protein to the cell membrane. The expression of SNAP(f)-ADRβ2 in the stable cell lines was confirmed by the reaction between BG-800 substrate and cell lysates. Microscopic examination confirmed that SNAP(f)-ADRβ2 was localized on the cell membrane. The signal intensity of the labeled cells was dependent on the BG-800 concentration. In vivo imaging study showed that BG-800 could be used to visualize xenograph tumors expressing SNAP(f)-ADRβ2. However, the background signal was relatively high, which may be a reflection of non-specific accumulation of BG-800 in the skin. To address the background issue, quenched substrates that only fluoresce upon reaction with SNAP-tag were synthesized and characterized. Although the fluorescence was successfully quenched, in vivo imaging with the quenched substrate CBG-800-PEG-QC1 failed to visualize the SNAP(f)-ADRβ2 expressing tumor, possibly due to the reduced reaction rate. Further improvement is needed to apply this system for in vivo imaging.     

Influence of neonatal treatment with porcine insulin (hormonal imprinting) on the receptor binding of insulins of porcine, rat and human origin

A single neonatal insulin treatment in the male rat decreases, whereas in the female rat it increases, the number of insulin receptors detected at the age of one month. No difference could be observed in affinity. The extent of the binding of rat, porcine and human insulin is different (the binding of human insulin is the most pronounced), meanwhile the alterations evoked by neonatal treatment are basically identical.     

Prolongation of skin xenograft survival with modified cultured fibroblasts

Cyclosporine A, one of the most potent immunosuppressive drugs, mediates some of its immunosuppressive and nephrotoxic effects by enhancing transforming growth factor-beta secretion and receptor expression. In this experimental study, the effect of cyclosporine pretreatment of cultured dermal fibroblasts on xenogeneic tissue rejection after microimplantation beneath skin grafts was investigated. The effects of the site-specific immunosuppressive strategy on skin xenograft survival were tested. Because the skin is an immunological indicator of the rejection of composite tissue allografts, it was considered that this strategy could be used as a supportive therapy for composite tissue allotransplantation in the future. In the first stage of the study, fibroblast cultures obtained from skin biopsy samples from five rats were treated with different single doses of cyclosporine (100 to 3000 ng/ml), and transforming growth factor-beta levels were measured in culture supernatants after 72 hours. In the second stage, 60 Sprague-Dawley rats were divided into six groups, as follows. For group I (sham), only the standard grafting procedure was performed. For group II, after the standard grafting procedure, rats were treated with intraperitoneal injections of 30 mg/kg (n = 5) or 10 mg/kg (n = 5) cyclosporine for 10 days. For group III, cultured fibroblasts obtained from skin biopsy samples from rats were treated with 100 or 500 ng/ml cyclosporine, and the cells were collected by light trypsinization and centrifugation after 72 hours. After the standard skin grafting procedure, modified fibroblasts were implanted between the graft and the recipient bed with a Pasteur pipet. For group IV, the same procedures as for group III were performed and then rats were treated with 10 mg/kg cyclosporine, administered intraperitoneally, for 10 days. For group V, in addition to standard grafting, unmodified fibroblasts (not treated with cyclosporine) were implanted between the graft and the recipient bed. For group VI, the same procedures as for group V were performed and then rats were treated with 10 mg/kg cyclosporine, administered intraperitoneally, for 10 days. The rejection process was observed macroscopically, and statistical significance was determined with the Mann-Whitney test (p < 0.01). Graft survival times were significantly prolonged in groups III and IV, compared with groups I, II, V, and VI (p < 0.001). No difference between groups III and IV was observed. The novel finding of this investigation is that xenogeneic skin graft survival times could be prolonged with microimplantation of cyclosporine-treated cultured fibro-blasts.     

Cytoplasmic proteins of porcine adipocytes: identification with monoclonal antibodies

In the present study, monoclonal antibodies were produced using porcine adipocyte extracts as the immunogen. Two of the monoclonal antibodies, designated CB6 and IB4, exhibited reactivity toward only cells containing lipid in stromal-vascular cell cultures. The antigens recognized by the CB6 and IB4 monoclonal antibodies were 50 kD and 55 kD proteins, respectively. In vivo, IB4 immunoreactivity was detected only in lipid-containing cells, whereas immunofluorescence using CB6 was also detectable around muscle fiber bundles underlying the subcutaneous mesenchyme. In fetal subcutaneous mesenchyme, CB6 and IB4 immunoreactivities toward lipid-containing cells increased with developmental age, but each was not detectable in cells containing the smallest lipid droplets. In stromal-vascular cultures containing adipocytes, 48 hour treatment with the anti-lipogenic agent, growth hormone, only slightly altered CB6 immunoreactivity, whereas IB4 immunoreactivity was reduced by more than sixfold. The exact identity of the CB6 and IB4 antigens was not determined, but each may be useful as markers for studying regulation of adipocyte metabolism.     

The structure of porcine parvovirus: comparison with related viruses

The structure of baculovirus-expressed porcine parvovirus (PPV) capsids was solved using X-ray crystallography and was found to be similar to the related canine parvovirus (CPV) and minute virus of mice (MVM). The PPV capsid protein has 57 % and 49 % amino acid sequence identity with CPV and MVM, respectively, but the degree of conservation of surface-exposed residues is lower than average. Consequently, most of the structural differences are on the surface and are the probable cause of the known variability in antigenicity and host range. The NADL-2 and Kresse strains of PPV have distinct tissue tropisms and pathogenicity, which are mediated by one or more of the amino acid residues 381, 386, and 436. These residues are on or near the surface of the virus capsid, where they are likely to be associated with virus-cell interactions.