L1 Region Research Articles

Multiple proteolytic events in caspase-6 self-activation impact conformations of discrete structural regions

Caspase-6 is critical to the neurodegenerative pathways of Alzheimer's, Huntington's, and Parkinson's diseases and has been identified as a potential molecular target for treatment of neurodegeneration. Thus, understanding the global and regional changes in dynamics and conformation provides insights into the unique properties of caspase-6 that may contribute to achieving control of its function. In this work, hydrogen/deuterium exchange MS (H/DX-MS) was used to map the local changes in the conformational flexibility of procaspase-6 at the discrete states that reflect the series of cleavage events that ultimately lead to the fully active, substrate-bound state. Intramolecular self-cleavage at Asp-193 evoked higher solvent exposure in the regions of the substrate-binding loops L1, L3, and L4 and in the 130s region, the intersubunit linker region, the 26-32 region as well as in the stabilized loop 2. Additional removal of the linker allowed caspase-6 to gain more flexibility in the 130s region and in the L2 region converting caspase-6 to a competent substrate-binding state. The prodomain region was found to be intrinsically disordered independent of the activation state of caspase-6; however, its complete removal resulted in the protection of the adjacent 26-32 region, suggesting that this region may play a regulatory role. The molecular details of caspase-6 dynamics in solution provide a comprehensive scaffold for strategic design of therapeutic approaches for neurodegenerative disorders.

Quantitative methylation analysis of human papillomavirus 16 L1 gene reveals potential biomarker for cervical cancer progression

Human papillomavirus 16 is the most prevalent type found in cervical cancer worldwide, accounting for >50% of all cases. Quantitative methylation analysis of human papillomavirus 16L1 gene within 5′ (CpGs 5600, 5606, 5609, 5615) and 3′ (7136 and 7145) regions to determine potential biomarker for cervical cancer progression was performed in exfoliated cervical cells collected from 101 Thai women of precancerous and cancerous lesions. Intermediate to high methylation levels (>20%) were detected in HPV16 5′L1 regions especially CpG 5600 of all cancerous (100%) and 50% of CIN3 samples, whereas normal/CIN1 samples (80%) showed methylation levels <20%. Our results indicate the potential use of HPV 16L1 gene methylation at specific site as a biomarker for prognostic cervical cancer screening, however, suitable cutoff should be further evaluated in a larger sample size.

Human papillomavirus 16L1-58L2 chimeric virus-like particles elicit durable neutralizing antibody responses against a broad-spectrum of human papillomavirus types.

The neutralizing antibodies elicited by human papillomavirus (HPV) major capsid protein L1 virus-like particle (VLP)-based vaccines are largely type-specific. An HPV vaccine inducing cross-neutralizing antibodies broadly will be cost-effective and of great value. To this end, we constructed HPV16L1-58L2 chimeric VLP (cVLP) by displaying HPV58 L2 aa.16-37 on the DE surface region of HPV16 L1. We found that vaccination with the HPV16L1-58L2 cVLP formulated with alum plus monophosphoryl lipid A (Alum-MPL) adjuvant elicited robust neutralizing antibodies in both mice and rabbits against all tested HPV types including HPV16/31/33/35/52/58 (genus α9), HPV18/39/45/59/68 (genus α7), HPV6/11 (genus α10), HPV2/27/57 (genus α4), and HPV5 (genus β1). Importantly, the cross-neutralizing antibody response was maintained at least 82 weeks in mice or 42 weeks in rabbits, and complete protection against HPV58 was observed at week 85 in mice. Our data demonstrate that HPV16L1-58L2 cVLP is an excellent pan-HPV vaccine candidate.

Translocation of the papillomavirus L2/vDNA complex across the limiting membrane requires the onset of mitosis.

The human papillomavirus type 16 (HPV16) L2 protein acts as a chaperone to ensure that the viral genome (vDNA) traffics from endosomes to the trans-Golgi network (TGN) and eventually the nucleus, where HPV replication occurs. En route to the nucleus, the L2/vDNA complex must translocate across limiting intracellular membranes. The details of this critical process remain poorly characterized. We have developed a system based on subcellular compartmentalization of the enzyme BirA and its cognate substrate to detect membrane translocation of L2-BirA from incoming virions. We find that L2 translocation requires transport to the TGN and is strictly dependent on entry into mitosis, coinciding with mitotic entry in synchronized cells. Cell cycle arrest causes retention of L2/vDNA at the TGN; only release and progression past G2/M enables translocation across the limiting membrane and subsequent infection. Microscopy of EdU-labeled vDNA reveals a rapid and dramatic shift in vDNA localization during early mitosis. At late G2/early prophase vDNA egresses from the TGN to a pericentriolar location, accumulating there through prometaphase where it begins to associate with condensed chromosomes. By metaphase and throughout anaphase the vDNA is seen bound to the mitotic chromosomes, ensuring distribution into both daughter nuclei. Mutations in a newly defined chromatin binding region of L2 potently blocked translocation, suggesting that translocation is dependent on chromatin binding during prometaphase. This represents the first time a virus has been shown to functionally couple the penetration of limiting membranes to cellular mitosis, explaining in part the tropism of HPV for mitotic basal keratinocytes.

A central region in the minor capsid protein of papillomaviruses facilitates viral genome tethering and membrane penetration for mitotic nuclear entry.

Incoming papillomaviruses (PVs) depend on mitotic nuclear envelope breakdown to gain initial access to the nucleus for viral transcription and replication. In our previous work, we hypothesized that the minor capsid protein L2 of PVs tethers the incoming vDNA to mitotic chromosomes to direct them into the nascent nuclei. To re-evaluate how dynamic L2 recruitment to cellular chromosomes occurs specifically during prometaphase, we developed a quantitative, microscopy-based assay for measuring the degree of chromosome recruitment of L2-EGFP. Analyzing various HPV16 L2 truncation-mutants revealed a central chromosome-binding region (CBR) of 147 amino acids that confers binding to mitotic chromosomes. Specific mutations of conserved motifs (IVAL286AAAA, RR302/5AA, and RTR313EEE) within the CBR interfered with chromosomal binding. Moreover, assembly-competent HPV16 containing the chromosome-binding deficient L2(RTR313EEE) or L2(IVAL286AAAA) were inhibited for infection despite their ability to be transported to intracellular compartments. Since vDNA and L2 were not associated with mitotic chromosomes either, the infectivity was likely impaired by a defect in tethering of the vDNA to mitotic chromosomes. However, L2 mutations that abrogated chromatin association also compromised translocation of L2 across membranes of intracellular organelles. Thus, chromatin recruitment of L2 may in itself be a requirement for successful penetration of the limiting membrane thereby linking both processes mechanistically. Furthermore, we demonstrate that the association of L2 with mitotic chromosomes is conserved among the alpha, beta, gamma, and iota genera of Papillomaviridae. However, different binding patterns point to a certain variance amongst the different genera. Overall, our data suggest a common strategy among various PVs, in which a central region of L2 mediates tethering of vDNA to mitotic chromosomes during cell division thereby coordinating membrane translocation and delivery to daughter nuclei.

Increased methylation of human papillomavirus type 16 DNA is associated with the severity of cervical lesions in infected females from northeast China.

Hypermethylation of the cytosine-phosphate-guanine (CpG) sites located at the 3'-major capsid protein L1 (3'L1) and the long control region (LCR) of the human papillomavirus (HPV) genome may be associated with the progression of cervical cancer (CC). However, the methylation status of the LCR of HPV type 16 DNA remains to be elucidated in an infected Chinese population. The aim of the present study was to investigate the association between methylation of the HPV 16 L1 gene and LCR, and the severity of cervical lesions in infected female patients. Therefore, bisulfite modification, polymerase chain reaction amplification and sequencing were used to analyze 122 HPV 16-positive clinical cervical swabs obtained from patients in northeastern China. The proportion of methylated samples at each of the 7 CpG sites within the 3'-L1/5'-LCR and 5 CpG sites within the promoter region was significantly increased in patients with CC, compared with that observed in high-grade squamous intraepithelial lesions (HSIL) and normal tissue/low-grade intraepithelial lesions (LSIL) (χ2 test, P<0.01). The mean methylation frequencies of the CpG sites 7,089 and 7,143 exhibited an area under the curve value of 0.822 [95% confidence interval (CI)=0.733-0.911] for distinguishing CC from other lesions, 0.787 (95% CI=0.700-0.874) for distinguishing normal/LSIL from HSIL and CC, and 0.763 (95% CI=0.652-0.874) for distinguishing CC from HSIL. These results suggest that the methylation of CpG sites within the HPV 16 3'-L1 and LCR region is correlated with the severity of cervical lesions. Quantification of HPV DNA methylation in the L1 gene and promoter region appears to provide a promising novel marker for distinguishing between normal tissue/LSIL, HSIL and CC in a Chinese population.

19F‐NMR Reveals the Role of Mobile Loops in Product and Inhibitor Binding by the São Paulo Metallo‐β‐Lactamase

AbstractResistance to β‐lactam antibiotics mediated by metallo‐β‐lactamases (MBLs) is a growing problem. We describe the use of protein‐observe 19F‐NMR (PrOF NMR) to study the dynamics of the São Paulo MBL (SPM‐1) from β‐lactam‐resistant Pseudomonas aeruginosa. Cysteinyl variants on the α3 and L3 regions, which flank the di‐ZnII active site, were selectively 19F‐labeled using 3‐bromo‐1,1,1‐trifluoroacetone. The PrOF NMR results reveal roles for the mobile α3 and L3 regions in the binding of both inhibitors and hydrolyzed β‐lactam products to SPM‐1. These results have implications for the mechanisms and inhibition of MBLs by β‐lactams and non‐β‐lactams and illustrate the utility of PrOF NMR for efficiently analyzing metal chelation, identifying new binding modes, and studying protein binding from a mixture of equilibrating isomers.

19 F-NMR Reveals the Role of Mobile Loops in Product and Inhibitor Binding by the São Paulo Metallo-β-Lactamase.

Resistance to β‐lactam antibiotics mediated by metallo‐β‐lactamases (MBLs) is a growing problem. We describe the use of protein‐observe 19F‐NMR (PrOF NMR) to study the dynamics of the São Paulo MBL (SPM‐1) from β‐lactam‐resistant Pseudomonas aeruginosa. Cysteinyl variants on the α3 and L3 regions, which flank the di‐ZnII active site, were selectively 19F‐labeled using 3‐bromo‐1,1,1‐trifluoroacetone. The PrOF NMR results reveal roles for the mobile α3 and L3 regions in the binding of both inhibitors and hydrolyzed β‐lactam products to SPM‐1. These results have implications for the mechanisms and inhibition of MBLs by β‐lactams and non‐β‐lactams and illustrate the utility of PrOF NMR for efficiently analyzing metal chelation, identifying new binding modes, and studying protein binding from a mixture of equilibrating isomers.

Molecular characterisation of fowl adenovirus type 7 isolated from poultry associated with inclusion body hepatitis in Poland

The fowl adenovirus field strain FAdV-JSN-5/10j (GenBank accession number KP879219) was isolated from the intestine of a 7-week-old chicken diagnosed with inclusion body hepatitis and simultaneously with Marek’s disease, and for that reason, it was chosen for molecular study. It was identified as fowl adenovirus genotype 7 (species Fowl aviadenovirus E) based on nucleotide sequence analysis of the loop L1 region of the hexon gene. Nucleotide sequence alignment of this strain, FAdV-7 reference strains B-3A ATCC VR-832 (AF339922) and YR36 (AF508955), and eight additional FAdV-7 field strains confirmed its classification as FAdV-JS-5/10j and showed that these viruses are very similar to each other. Additionally, we described mutations and their influence on the amino acid sequence, nucleotide composition, and relative synonymous codon usage. Immunofluorescence of cell cultures infected with 104.5 TCID 50 per 0.1-ml dose of the FAdV-JSN-5/10j strain demonstrated the presence of a cytopathic effect. Infection of fowl with adenoviruses raises concerns for poultry production, and thus, the efficient detection of adenovirus infection is crucial. This is the first attempt to describe the molecular characteristics of FadV-7 strains isolated in Poland.

Prevalence and Determinants of Oral Human Papillomavirus Infection in 500 Young Adults from Italy.

Although the prevalence of human papillomavirus (HPV)-related oropharyngeal squamous cell carcinoma (OPSCC) is increasing in developed countries and becoming a relevant health issue, the natural history of oral HPV infection is still unclear. Estimating the infection’s prevalence in specific populations and identifying risk factors can widen our understanding of its natural history and help to delineate appropriate prevention strategies. This study sought to (i) determine oral HPV prevalence and genotype distribution in a large series of young Italian adults, (ii) validate an oral rinse sampling/storage protocol, and (iii) pinpoint factors associated with oral HPV infection. Five hundred students, nurses, and technicians (19–35 years-old) studying and working at/for the University of Padua were recruited. Each participant was provided with an oral rinse sampling kit and instructions for use. They were also asked to complete an anonymous questionnaire concerning their demographic characteristics and behaviors. The questionnaires and oral rinse containers were labeled with the same identification code number. The oral rinse samples were tested using a bead-based multiplex BSGP5+/6+-MPG genotyping assay which amplifies the L1 region of 51 mucosal HPV types. The prevalence of oral HPV infection was 4.0% (95% confidence interval (CI), 2.5%-6.1%); those of 14 high-risk HPV types and of HPV-type 16 (HPV16) infection were 2.2% (95% CI, 1.1%-3.9%) and 1.6% (95% CI, 0.6%-3.1%), respectively. HPV16 was the most frequent genotype (40.0% of oral HPV infections). No association was found between oral infection and the co-variables studied (gender, tobacco, alcohol and illegal drug use, number of sex and oral sex partners, HPV vaccination status, history of HPV and sexually transmitted infections, abnormal pap smears, recurrent tonsillitis and tonsillectomy). The oral rinse sampling protocol outlined here proved to be simple, efficient and well tolerated, and the prevalence rate can be considered reliable and thus useful to guide future research. Determinants of oral HPV infection are still unclear and further studies are certainly warranted.

Humans Papillomavirus (Hpv) Infections in Female Sex Workers in Cote D’ivoire

Background: Human Papillomaviruses (HPV) are small virus non-enveloped double-stranded circular DNA. They infect epithelial cells (mucous membranes and skin). Many studies have indicated that having multiple sexual partners may lead to higher HPV transmission. Thus female sex workers (FSWs) may be at greater risk of infection compared to the general population. In female sex workers (FSWs), the risk of HPV infection and cervical cancer is especially high. The aim of this work is to determine the prevalence and the genotypes of Humans Papillomavirus (HPV) that circulate in female sex workers populations in Cote d’Ivoire. Methods: From December 2015 to May 2016, cervical samples from 350 female sex workers were tested for some HR-HPV. HPV DNA was amplified using PGMY09 /11 primers which generated 450 base pairs at the L1 region. The samples harboring HPV DNA were genotyped using the multiplex PCR with HPV 16, 18, 31, 33, 35, 45 and 51 primers. Results: The mean age of this population was 32.5 years. On 350 female sex workers HPV DNA was obtained in 51.5% of the population. A total of 168 (94.38%) specimens harboring HPV DNA were genotypes using multiplex PCR versus 5.61 %, which were not genotyped using HPV 16,18, 31, 33, 35, 45 and 51 by multiplex PCR. These 168 strains permit us to identify 204 strains of HPV on whom we have 88.69 % with single infection while 11.30 % with a multiple infection. Among the multiple infection 36.84 % had respectively double and triple HPV infection and 26.31 % had four HPV infections. HPV genotypes prevalence was the followed: HPV 16 (22.47%), HPV 18 (26.97%), HPV 35 (11.23%), HPV 31 and HPV 33 (7.86%) respectively and HPV 45 (7.30%). Any case of HPV genotype 51 was founded. Conclusion: The prevalence of HPV infection in female sex workers is high. The most genotypes which circulate in female sex workers are type 16 and type 18 which are the best known in the world.

Association between human papillomavirus infection and lung cancer

Objective: To investigate the association between human papillomavirus (HPV) and lung cancer. Methods: We examined a series of 83 lung cancer patients with HPV DNA in both lung tumor specimens and adjacent normal specimens from Fujian province. Twenty-one of the most clinically relevant HPV types from the highly conserved L1 region of the viral genome were analyzed, using the PCR amplification and were followed by reverse hybridization with specific probes. Chi-square test of paired design was used to test the difference of HPV positive rates between lung cancer specimens and adjacent normal specimens. Chi-square test and Fisher's exact test were used to analyze the differences of HPV positive rate of tumor specimens on factors as gender, age, histological subtype, clinical stage, smoking status and alcohol consumption. Results: HPV was detected in 7 of the 83 tumor specimens and in 6 of the paired normal lung tissues. There was no significant correlation between HPV and lung cancer (P>0.999). Neither demographic characteristics nor clinical features were found with significant differences on HPV in lung cancer tissues (P>0.05). Conclusion: Our data showed that HPV was not significantly associated with the risk of lung cancer in Fujian province.

Emulsion-mediated synthesis of hierarchical mesoporous-macroporous Al-Mg hydrotalcites

A new sol-gel method mediated by the use of paraffin emulsions is presented for the synthesis of hierarchical mesoporous-macroporous Al-Mg hydrotalcites. Prior to the synthesis, the construction of ternary diagrams for paraffin/surfactant/ethanol helped to identify the most suitable compositional synthesis domain. Two distinct regions, denoted L1 (transparent microemulsion) and L2 (opaque emulsion) were identified for low and high concentrations of paraffins, respectively. The ternary diagrams revealed some features: i) n-hexane (C6) had the smallest L2 region; ii) the high viscosity of n-octadecane (C18) did not allow control of phase separation, and iii) the most suitable paraffin for the use as the emulsifier was the intermediate-sized n-dodecane (C12). The use of paraffin in concentrations included in the L1 region did not contribute to the creation of pores, but the emulsion templating in the L2 region led to hierarchical porous hydrotalcites when were calcined up to 250 °C having enhanced textural properties and presenting mesopores and macropores with mean sizes of 0.02 and 1 μm, respectively. The slit-like shaped mesopores were caused by the partial exfoliation of hydrotalcite layers unordered packed around the emulsion droplets, while the macropores arose from voids left by the droplets after thermal treatment. The singular textural properties of the studied Al-Mg hierarchical porous hydrotalcites turns the emulsion mediated-synthesis a potential method to obtain more efficient catalysts to be used in process involving bulky molecules.

Prevalencia de tipos de virus del papiloma humano en hombres que tienen sexo con hombres, en Chihuahua, México

The aim of this work was to determine human papilloma virus (HPV) high and low risk (HR and LR) genotypes prevalence, and other HPV infection associated risk factors, in men who have sex with men (MSM). A cross-sectional study was conducted, to this end, 104 MSM responded a social demographic and sexual behavior questionnaire and consented sampling of anal epithelium for molecular analysis by polymerase chain reaction (PCR), with β -globin primers as internal control, and HPV L1 region primers. Subsequently, typing was performed by sequencing. Prevalence of HPV in MSM was 34%: 10% HR and 24% LR. The most frequent oncogenic HPV types were 16 (20%), 73 (20%) and 45 (20%), and non-oncogenic VPH 11 (36%), 62 (28%) and 6 (12%). MSM less than 30 years old had greater HPV prevalence. Presence of human immunodeficiency virus (HIV) was not significantly associated to HPV infection in the studied population.

899P - Defining the genomic landscape of vulvar squamous cell carcinoma (VSCC) using next generation sequencing: the role of HPV infection

Surgery for vulvar squamous cell carcinoma (VSCC) is associated with physical and psychological morbidity. Targeted therapies have not influenced outcomes, in part because the genomic landscape of VSCC remains incompletely understood and is complicated by two etiologies: HPV-dependent and HPV-independent disease. Next-generation sequencing (NGS) was used to identify somatic mutations by HPV status (+/-). We conducted a retrospective review of patients (pts) seen at the Ottawa Hospital, Ottawa, Canada between 2000-2012 for VSCC. Clinical and pathological data was collected and pts with adequate tumor for HPV and molecular analysis were identified. HPV status was determined by polymerase chain reaction (PCR) with primers targeting the L1 region of HPV DNA. The Ion AmpliSeq™ Cancer Hotspot Panel v2 is being used to examine the presence of 50 known mutations. Forty-seven pts with VSCC were included. 22/47 (47%) were HPV+ and 25/47 (53%) were HPV-. Median age at diagnosis was 60 years (IQR 49-76) and 69 years (IQR 58-77), for HPV+ and HPV-, respectively. Disease stage by HPV status (+ vs -) was: I (15 vs 16), II (2 vs 0), III (2 vs 9) and IV (3 vs 0). Molecular data is available on 31 pts (17 HPV + , 14 HPV-). Among HPV+ pts, 15/17 had ≥ 1 mutation. Mutational frequencies among HPV+ pts were: TP53 7/17, PIK3CA 6/17, KDR 5/17, KIT: 3/17, FGFR3 3/17, and one mutation each of PTEN, CTNNB1, APC, KRAS, ERBB4, ATM, SMARCB1, FLT3, CDK2A. Among the HPV- pts, 12/14 had ≥ 1 mutation. Mutational frequencies among HPV- pts were: TP53 8/14, HRAS 3/14, CDKN2A 2/14, PI3KCA 2/14, KDR 1/14 and GNA11 1/14. VSCC is characterized by a high mutation rate and a high prevalence of HPV infection. HPV-dependent and HPV-independent disease have unique mutational profiles. The high prevalence of “actionable” mutations supports the need for trials of targeted therapies. Molecular data on the full study cohort (n = 47) will be presented.

Micellar crowding and branching in a versatile catanionic system

The catanionic system didodecyldimethylammonium bromide (DDAB)-sodium taurodeoxycholate (STDC)-D2O is characterized by an exceptionally extended L1 region. The comparison of self-diffusion coefficient of the solvent and the DDAB embedded in the micelles provided information about hydration of the aggregates. Moreover, correlating self-diffusion and 14N NMR relaxation measurements new insight could be obtained regarding the translational and rotational micellar motions in the crowded solutions of systems with 0.2 DDAB-STDC molar ratio. 1H 2D NMR spectra gave some hints about the mutual arrangement of DDAB and STDC within the aggregates. For samples with 1.8 and 2.6 DDAB/STD molar ratios the 14N data were in agreement with the presence of somewhat branched, interconnected micelles. 23Na and 81Br dynamic parameters resulted particularly sensitive to the surrounding environment.The peculiar rheological behaviour of the, highly concentrated, branched micelles samples, namely the steady oscillations in the step tests, was found to be an example of instability originated by the combined effect of material elasticity and slippage at the fixed wall.

Prevalence, Genotype Distribution and Risk Factors for Cervical Human Papillomavirus Infection in the Grand Tunis Region, Tunisia.

Implementation of Human Papillomavirus (HPV) vaccination should be considered a key cervical cancer prevention strategy in Tunisia, where Pap smear screening is not efficient. This study aims to estimate the prevalence and to identify risk factors associated with HPV infection among women from Grand Tunis, Tunisia. We conducted a cross-sectional study, between December 2012 and May 2013. Eligible women for this study were those aged 18–65 years, sexually active, who sought medical attention at their primary health care centre or clinic in Grand Tunis, Tunisia and who gave written consent. A liquid-based Pap smear sample was obtained from all women using a cervical brush. Only women with betaglobin positive test were further analysed for HPV detection and typing. A nested-PCR of the L1 region was performed followed by reverse line blot hybridization to facilitate the specific detection of 31 HPV genotypes. Multiple logistic regression modeling was used for the analysis of associations between variables with some considered possible confounders after checking for interactions. A total of 391 women were enrolled in this study and 325 out of the 391 cervical samples were positive for the betaglobin test. Overall HPV prevalence was 13.2% [9.8%−17.5%], with the following most prevalent HPV genotypes: HPV6 (40%), HPV40 (14%), HPV16 (12%), HPV52 (9%), HPV31 and HPV59 (7%), followed by HPV68 (4%). Mean age of HPV positive women was 40.7±0.92 years. Independently associated risk factors of HPV infection were smoking (OR:2.8 [0.8–9.6]), low income (OR:9.6 [1.4–63.4), bad housing type (OR:2.5 [1–6.8]), partner with multiple sexual relationship (OR:4.5 [0.9–22.9]) and single women (widowed, divorced, separated, never married) (OR:6.9 [1.1–42.2]). This study provides the first national-based estimate of HPV prevalence in Tunisia. Our findings contribute to the evidence on the current burden of HPV infection, the critical role of sexual behaviour and socioeconomic status and call for increased support for the screening program in Tunisia to prevent cervical cancer. These results allow us to evaluate the cost-effectiveness of vaccine program implementation in Tunisia in future.

High prevalence of abdominal aortic calcification in patients with primary hyperparathyroidism as evaluated by Kauppila score.

The prevalence of abdominal aortic calcification (AAC) in primary hyperparathyroidism (PHPT) is unknown. We assessed both prevalence and severity of AAC in PHPT postmenopausal women. In this study 70 PHPT postmenopausal women and 70 age- and sex-matched controls were enrolled. Each participant underwent biochemical evaluation, lateral spine radiograph, bone mineral density (BMD) measurement (lumbar, femoral, radial sites), and kidney ultrasound. Lateral lumbar films were analyzed in the region of L1-L4 vertebrae and the Kauppila score (a semi-quantitative grading system) was used to assess the severity of AAC. There were no differences regarding demographic and cardiovascular risk factors in the two groups. PHPT patients had higher prevalence of kidney stones (30% vs 7%, P=0.0008) and lower radial BMD values (0.558±0.071 vs 0.588±0.082 g/cm(2), P<0.05) compared with controls. PHPT patients showed higher prevalence of AAC (31 vs 18, P=0.03), with more severe calcifications (Kauppila score 7.35±6.1 vs 5.05±3.5, P=0.007). PHPT patients with AAC were older and had been suffering from the disease for a longer period compared with those without ACC. Moreover, PHPT patients with severe AAC had mean higher serum parathyroid hormone levels compared with patients with moderate or mild calcifications. In PHPT patients with AAC, multiple regression analysis, adjusted for age and years since diagnosis, showed that only parathyroid hormone significantly correlated with Kauppila score. We found a higher prevalence and severity of AAC in PHPT related to parathyroid hormone effect.

Properties and ionic self-assembled structures from mixture of a bola-type strong alkali dication and a branched phosphoric acid

Ionic self-assembled structures have been prepared successfully between di-(2-ethylhexyl) phosphoric acid (DEHPA) and hexamethonium hydroxide (HMO(OH)2). The DEHPA/HMO(OH)2 complexes show good surface activity at a wide mixing molar ratio of DEHPA to HMO(OH)2 (ρ), within which the critical micellar concentration (cmc) is far below that of any single component. In bulk aqueous solutions, rich phase behavior was observed by varying cDEHPA and cHMO(OH)2. When the concentration of HMO(OH)2 is in the range of 10–100mmolL−1, isotropic L1 phases, birefringent Lα phases and a phase-separated region were successively observed with increasing cDEHPA. At high cHMO(OH)2 range (>78mmolL−1), a narrow L1/Lα two-phase region with the Lα phase at the bottom was noticed between the single L1 and single Lα phase regions. The rheological properties of the samples in the single Lα phase region at 2.6⩽ρ⩽2.8 are quite similar. Cryo-TEM and freeze-fracture TEM (FF-TEM) observations revealed the presence of multilamellar vesicles with flexible and even branched bilayers. At 2.2⩽ρ⩽2.6, however, the rheological properties are highly sensitive to ρ due to the sophisticated self-assembly behavior as proved by imaging studies and 2H NMR measurements. Closely-stacked flat structures which look like foams or cellular networks have been newly discovered. Interestingly, NaCl could arouse an L1 to Lα phase transition due to the suppression of the effective area of the hydrophilic headgroups of the ionic complexes, leading to an increase of the critical packing parameter p. The viscoelasticity properties of the salt-containing Lα phases decreased with increasing salinity. We hope our research can provide new ideas for the construction of supramolecular materials by surfactant ionic self-assembly (SISA) strategy.

An Unusual Finding of Buschke-Lowenstein Tumor Associated with 6, 39 and 53 HPV Genotypes in a Young Immunocompetent Female

Background: Buschke-Lowenstein tumor (BLT) or giant condyloma acuminatum (GCA), is a rare disease caused by Human Papilloma virus (HPV) types 6 and 11, more common in immunocompromised males. We describe a rare case of BLT associated with mixed low and high-risk HPV type’s infection in a very young immunocompetent female. Methods: On August 2012, an 18-year old female was admitted to the Unit of Obstetrics and Gynecology, with a 3-4 month history of small excrescences in the perineal/vulvar area which had rapidly increased in size. A diagnosis of BLT was suspected; an incisional biopsy was carried out. On the cervical and vulvar cytobrush samples, Polymerase Chain Reaction (PCR) of specific sequences of the HPV genome L1 region was performed. Surgical treatment was carried out by cold knife excision. Results: Clinically, the tumor presented as exophytic white lesion, characterized by an irregular surface with acuminate excrescenses. Microscopic images are consistent with diagnosis of Buschke-Lowenstein tumor. Performed PCR revealed the presence of HPV types 6, 39 and 53. After surgery the patient showed no signs of recurrence during the 4 years of follow-up. Conclusions: This report a rare case of BLT in a young immunocompetent female, associated with multiple HPV type infection. The studied case suggests that pathologies with a BLT-like clinical and histological presentation and mixed HPV genotypes can be found in young immunocompetent patient.