Calcineurin inhibitors and male fertility after renal transplantation - a review.

Renal transplantation and restoration of renal function are associated with significant favourable changes regarding the reproductive capacity of male patients with previous end-stage renal disease. However, there is evidence that some of the immunosuppressive agents may impair male fertility after all. Calcineurin inhibitors (CNIs), cyclosporine A and tacrolimus (FK506), which constitute the cornerstone of immunosuppression regimen following renal transplantation, have been implicated in causing an overall decline in the fertilisation capacity of male renal transplant recipients (RTRs). In this review, data from human clinical studies are collectively presented in an effort to estimate the potential adverse effects of CNIs on the masculine reproductive organs, the hormonal axis of males, the process of spermatogenesis and generally the male RTRs capacity to fertilise.

Active terahertz beam steering by photo-generated graded index gratings in thin semiconductor films.

We demonstrate active beam steering of terahertz radiation using a photo-excited thin layer of gallium arsenide. A constant gradient of phase discontinuity along the interface is introduced by an spatially inhomogeneous density of free charge carriers that are photo-generated in the GaAs with an optical pump. The optical pump has been spatially modulated to form the shape of a planar blazed grating. The phase gradient leads to an asymmetry between the +1 and -1 transmission diffracted orders of more than a factor two. Optimization of the grating structure can lead to an asymmetry of more than one order of magnitude. Similar to metasurfaces made of plasmonic antennas, the photo-generated grating is a planar structure that can achieve large beam steering efficiency. Moreover, the photo-generation of such structures provides a platform for active THz beam steering.

The role of pelvic bone anatomy in the pathogenesis of inguinal hernia.

BACKGROUND: a small number of reports have suggested that inpatients with inguinal hernia, certain pelvic anatomical traits prevail, which are characterized by the low position of the groin, resulting in a marked verticality of the inguinal fold.Based on this notion we investigated a possible correlation of this anatomical characteristic with hernia development, by applying a simple clinical research protocol. METHODS: two groups were formed. A study group including 25 adult patients with a history of inguinal hernia or aclinically proven inguinal hernia and a control group of 10 individuals of the same age and sex distribution with the study group, but with no history of inguinal hernia. On anteroposteriorpelvic plain x-rays, we measured Radoievitch'€™s angle which is formed by the interspinal line and the line passing from the anterior superior iliac spine and the pubic tubercle.Mean values of Radoievitch'€™s angle were then compared between the two groups. RESULTS: patients with inguinal hernia had greater values concerning Radoievitchâ€'s angle in a statistically significant manner when compared to controls (p=0.004). CONCLUSIONS: individual variability in the configuration of the adult bony pelvis is an important factor in the development of inguinal hernia. Pelvimetry could affect the choice of hernia repair and radiological pelvimetry could be included in the preoperative planning of hernia repair.

Prediction of complicated appendicitis risk in children.

OBJECTIVE: We aimed to predict the risk of complicated appendicitis in children, constructing a risk-based prediction tool with the optimal combination of sensitivity and specificity outcomes. PATIENTS AND METHODS: This is a prospective study on a random sample of children with acute appendicitis who underwent appendectomy. Clinical examination, history, routine laboratory tests, Alvarado and pediatric appendicitis scores, operative and histopathological findings were taken into consideration. The predictive ability of the outcome variables was assessed by the Receiver Operating Characteristics (ROC) analysis. The overall predictive ability and determination of the best cut-off value (the higher sum of sensitivity plus specificity) were calculated. A Classification and Regression Tree (CRT) was used to create a multi-level classification algorithm. The model was set to predict the outcome of complicated appendicitis, considering as potential predictors the demographic characteristics, the clinical findings, and the outcome parameters. RESULTS: The various combinations of clinical and laboratory parameters did not improve their overall diagnostic ability. However, the CRT analysis resulted in a short classification algorithm based on the Pediatric appendicitis score, neutrophils percentage and the CRP. This model yielded a significantly better predictive ability than all the other combinations of the outcome parameters. The application of the model would predict complicated appendicitis with 90% sensitivity and 78.6% specificity. CONCLUSIONS: The constructed predictive model may be a useful tool for daily practical use by the clinician, especially in areas where modern diagnostic imaging facilities are absent or not always available. Clinical evaluation and close follow-up remain the more accurate preoperative method to decide the performance and timing of appendectomy.

Effect of root canal irrigant (sodium hypochlorite & saline) delivery at different temperatures and durations on pre-load and cyclic-loading surface-strain of anatomically different premolars.

AIM: To evaluate the effect of NaOCl (5%) and saline (control) irrigant delivery at different temperatures and durations on pre-load and cyclic-loading tooth-surface-strain (TSS) on anatomically different premolars. METHODOLOGY: Single-rooted premolars (n = 36), root-canal-prepared in standard manner, were randomly allocated to six irrigation groups: (A1) NaOCl-21 °C; (A2) NaOCl-60 °C; (A3) saline-21 °C then NaOCl-21 °C; (A4) saline-60 °C then NaOCl-21 °C; (A5) saline-21 °C then NaOCl-60 °C; (A6) saline-60 °C then NaOCl-60 °C. A1-2 received nine 10-min irrigation periods (IP) with NaOCl; A3-6 received nine 10-min IP with saline, followed by 9 IP with NaOCl at different temperature combinations. Premolars (n = 56) with single, fused or double roots prepared by standard protocol, were stratified and randomly allocated to: (B1) saline-21 °C; (B2) saline-80 °C; (B3) NaOCl-21 °C; (B4) NaOCl-80 °C. TSS (µÑ”) was recorded pre-irrigation, post-irrigation and pre-load for each IP and during cyclic loading 2 min after each IP, over 30-274 min, using strain-gauges. Generalised linear mixed models were used for analysis. RESULTS: Baseline TSS in double-rooted premolars was significantly (p=0.001) lower than in single/fused-rooted-premolars; and affected by mesial-wall-thickness (p=0.005). There was significant increase in loading-TSS (µÑ”) after NaOCl-21 °C irrigation (p=0.01) but decrease after NaOCl-60 °C irrigation (p=0.001). TSS also increased significantly (p = 0.005) after Saline-80 °C irrigation. Pre-load "strain-shift" was noted only upon first saline delivery but every-time with NaOCl. Strain-shift negatively influenced loading-TSS after saline or NaOCl irrigation (A3-6) but was only significant for saline-21 °C. CONCLUSIONS: Tooth anatomy significantly affected its strain characteristics, exhibiting limits within which strain changes occurred. Intra-canal introduction of saline or NaOCl caused non-random strain shifts without loading. Irrigation with NaOCl-21 °C increased loading tooth strain, as did saline-80 °C or NaOCl-80 °C but NaOCl-60 °C decreased it. A "chain-link" model was proposed to explain the findings and tooth biomechanics.

Affinity maturation of an anti-V antigen IgG expressed in situ through adenovirus gene delivery confers enhanced protection against Yersinia pestis challenge.

Genetic transfer of neutralizing antibodies (Abs) has been shown to confer strong and persistent protection against bacterial and viral infectious agents. Although it is well established that for many exogenous neutralizing Abs increased antigen affinity correlates with protection, the effect of antigen affinity on Abs produced in situ after adenoviral gene transfer has not been examined. The mouse IgG2b monoclonal Ab, 2C12.4, recognizes the Yersinia pestis type III secretion apparatus protein, LcrV (V antigen), and confers protection in mice when administered as an IgG intraperitoneally or after genetic immunization with engineered, replication-defective serotype 5 human adenovirus (Ad). The 2C12.4 Ab was expressed as a single-chain variable fragment (scFv) in Escherichia coli and was shown to display an equilibrium dissociation constant (K(D))=3.5 nM by surface plasmon resonance analysis. The 2C12.4 scFv was subjected to random mutagenesis, and variants with increased affinity were isolated by flow cytometry using the anchored periplasmic expression bacterial display system. After a single round of mutagenesis, variants displaying up to 35-fold lower K(D) values (H8, K(D)=100 pM) were isolated. The variable domains of the H8 scFv were used to replace those of the parental 2C12.4 IgG encoded in the Ad vector, AdalphaV, giving rise to AdalphaV.H8. The two adenoviral vectors resulted in similar titers of anti-V antigen Abs 3 days after immunization, with 10(9), 10(10) or 10(11) particle units (pu). After intranasal challenge with 363 LD(50) (lethal dose, 50%) of Y. pestis CO92, 54% of the mice immunized with 10(10) pu of AdalphaV.H8 survived through the 14 day end point compared with only 15% survivors for the group immunized with AdalphaV expressing the lower-affinity 2C12.4 (P<0.04; AdalphaV versus AdalphaV.H8). These results indicate that affinity maturation of a neutralizing Ab delivered by genetic transfer may confer increased protection not only for Y. pestis challenge but also possibly for other pathogens.

Prognostic and predictive factors of invasive ductal breast carcinomas.

PURPOSE: To investigate the significance of certain immunohistochemical markers, namely estrogen (ER) and progesterone receptors (PgR), c-erbB-2 oncogene, p53 tumor suppressor gene and E-cadherin adhesion molecule, in invasive ductal breast carcinomas. METHODS: A series of 102 primary breast carcinomas of the ductal type and a standard immunohistochemical technique was used to detect the aforementioned biological markers. The findings were related to various clinical and pathological tumor characteristics, including lymph node metastases. RESULTS: ER and E-cadherin were expressed more commonly in tumors of low histological grade and small number (< or =3) of metastatic lymph nodes, whereas c-erbB-2 and the p53 gene were usually expressed in breast tumors of high histological grade and increased number (>3) of metastatic lymph nodes. PgR, on the other hand, was detected frequently in patients with early menarche and metastases in <3 lymph nodes, but this tendency was not statistically significant. CONCLUSION: The use of these biomarkers, preferably in combination, may provide additional prognostic and therapeutic information which may be proved useful in planning breast cancer treatment.

Touch imprint cytological diagnosis of nodal Langerhans cell histiocytosis.

Touch imprint cytological diagnosis of nodal Langerhans cell histiocytosis. Langerhans cell histiocytosis (LCH) is a rare neoplasm of the mononuclear phagocytic immunoregulatory system of unknown aetiology. Nodal involvement is uncommon. Cytological findings have seldom been described. A case study of LCH, arising in a submandibular node of a 42-year-old female, is reported. Fine needle aspiration smears were highly cellular and composed of a mixed cell population including eosinophils, lymphocytes, neutrophils, and macrophages. Imprint slides from the surgical specimen of the excised node exhibited Langerhans cells with nuclear grooves, leading to a diagnosis suggestive of LCH. Immunohistochemical staining of the node sections with CD1a and S-100 confirmed this diagnosis. In conclusion, cytology may favorably contribute to the diagnosis of LCH.

Accessory parotid gland carcinoma ex pleomorphic adenoma. Case study diagnosed by fine needle aspiration.

OBJECTIVE: The accessory parotid gland is salivary tissue separated from the main parotid gland and lying on masseter muscle. It has secondary duct emptying into the Stensen's duct. The accessory parotid gland exists in 21-61% of individuals. However, the appearance of an accessory parotid tumor is rare, with a reported frequency of 1-7.7% of all parotid gland tumors. Carcinoma ex pleomorphic adenoma arises from a pre-existing benign mixed tumor. Most of these tumors will have malignant epithelial component, but not malignant stromal component. Reports of Fine Needle Aspiration Cytological (FNAC) diagnosis of malignant mixed tumor are uncommon and have been limited to cases arising in the parotid. We report a case of carcinoma ex pleomorphic adenoma of the accessory lobe of the parotid, and address the cytopathology features and pitfalls of this condition. CASE: A 73 aged female presented with a right nontender midcheek mass. The lesion had been present 18 months, with a recent increase in size. FNA was performed and the smears demonstrated features indicative of pleomorphic adenoma admixed with findings indicative of a poorly differentiated carcinoma. CONCLUSION: FNAC can accurately diagnose carcinoma ex pleomorphic adenoma when strongly fixed requirements are implemented.

Long-term bone marrow cultures (LTBMC) from essential thrombocythemia (ET) patients with or without JAK2617V>F mutation.

Approximately half of essential thrombocythemia (ET) patients and almost all with polycythemia vera (PV) bear the activating JAK2617V>F point mutation, which arises at the multipotent haemopoietic progenitor cell level. Although ET is mainly characterized by megacaryocyte proliferation, the cases that are positive for the JAK2617V>F mutation also show increased bone marrow cellularity and higher erythrocyte and granulocyte counts. After establishing short- and long-term bone marrow cultures we found that the frequency of committed haemopoietic progenitors in the bone marrow, was not increased in JAK2617V>F positive ET compared to the negative ones, whereas in long-term cultures (LTBMC) JAK2617V>F positive ET display a growth pattern more similar to that observed in LTBMC produced by PV marrow cells. Our data support the notion that JAK2617V>F positive ET and PV represents a continuum spectrum of alterations within the same disease.

Intraoperative touch imprint cytological analysis of sentinel lymph nodes for the presence of metastases in malignant melanoma.

AIM: Sentinel lymph node (SLN) biopsy has revolutionized lymph node staging in patients with malignant melanoma. Intraoperative evaluation is a new addition to the SLN procedure that allows for a one-step regional lymph node dissection to be performed when the SLN biopsy findings are positive. The discriminatory immunostaining pattern with the S-100 and HMB45 monoclonal antibodies allows intraoperative immunocytochemical evaluation of imprint smears of SLNs for melanoma metastases. METHODS: One hundred twenty eight SLNs from a cohort of 52 patient-cases that had been identified using sulfur colloid as a radioactive tracer and isosulfan blue were bisected for rapid Diff-Quick stained touch preparations. Intraopera-tive evaluation of sentinel node status by imprint cytology was correlated with the histopathological results of permanent sections. Tumor-negative nodes in routine paraffin sections were further investigated with the employment of the S-100 and HMB45 antibodies. RESULTS: Thirty-six of all SLNs harbored metastases in paraffin sections, from which 32 were identified by imprint cytology (sensitivity 88.8%). Three SLNs were positive by imprint cytology and negative by histopathology of paraffin sections. Comparison of the results of the touch preparations with the final histopathology (hematoxylin-eosin and S-100/ HMB45 stains) demonstrated a sensitivity of 83.3% and a negative predictive value of 92.5%. The specificity and positive predictive value were 100% respectively. CONCLUSION: Touch imprint cytology is potentially useful for intraoperative evaluation of SLNs in malignant melanoma patients. Results can be improved if the surface sampled is appropriately enlarged and a rapid immunohistochemical S-100/HMB45 stain on the imprints is utilized.

Fibroadenoma masquerading carcinoma on fine-needle aspiration of the breast.

OBJECTIVE: Benign and malignant lesions of the breast may have similar appearances on fine-needle aspiration cytology. We report a case of fibroadenoma that was diagnosed as carcinoma by cytology. CASE STUDY: Breast fine-needle aspiration biopsy was highly cellular and composed of bland-appearing spindle/columnar cells that could represent either epithelial or stromal cells; the case was reported as positive and the patient had subsequent excisional biopsy taken. RESULTS: On microscopic examination, smears were hypercellular and had many single cells and clusters of columnar/ elongate cells No obvious bipolar cells of myoepithelial origin were seen. Significant atypia was noted. Immunocytochemistry for smooth muscle actin was not performed due to insufficient material. CONCLUSIONS: Some cases of fibroadenoma and carcinoma can be very difficult to distinguish on fine needle aspiration cytology smears. Immunocytochemistry may be of help if sufficient material is provided. To avoid false positive diagnosis on cytology, it is best to report such a case as intermediate (atypical/suspicious) with final interpretation pending excisional biopsy.

Neck nodal metastases from unknown primary: case series.

BACKGROUND: Neck nodal metastases from occult primary constitute about 5%-10% of all hosts harboring carcinoma of unknown primary site. Metastases in the upper and middle neck (levels I-II-III-IV-V) are generally attributed to head and neck cancers, whereas the lower neck (level IV) involvement is often associated with primaries below the clavicles. Diagnostic procedures include a careful clinical evaluation and a fiberoptic endoscopic examination of the head and neck mucosa, biopsies from all suspicious sites or blindly from the sites of possible origin of the primary, computerized tomography (CT) scan, and magnetic resonance radiology (MRI). The most frequent histological finding is squamous cell carcinoma, particularly when the upper neck is involved. SETTINGS: We report three cases of patients presented with nodal metastases of the neck from unknown primary site and we also describe the diagnostic and therapeutic approach employed in each one. RESULTS: One patient harbored a neuroendocrine metastatic deposit, the second patient a poorly differentiated carcinoma and the third one a malignant melanoma. CONCLUSIONS: Diagnostic procedures should be aimed at clarifying the histology of the nodal metastases and detecting the primary tumor site.

Endometriosis involving the rectus abdominis muscle and subcutaneous tissues: fine needle aspiration appearances.

OBJECTIVE: Endometriosis is defined as functioning endometrial tissue outside the uterine cavity. It occurs in up to 15% of menstruating females and in most cases is located within the pelvis. Endometrial implants, however have been described in soft tissues, particularly in the skin and subjacent tissues of surgical scars, and diagnosis might be problematic. CASE STUDY: A 32 aged female presented with a suprapubic abdominal mass, which appeared suddenly after exercise. Fine needle aspiration was performed. RESULTS: Epithelial sheets were shown in direct aspirates. No evident endometrial stromal cells were seen. CD10 immunostaining in additional cell block preparations using a commercial antibody gave positive results. The cell pattern and immunocytochemical profile suggested a cytodiagnosis of endometriosis. The patient was administered with leuprolide acetate. She experienced adverse effects related to estrogen deficiency. Medical treatment was discontinued and the patient underwent surgical excision. Histological sections revealed endometrial glands surrounded by stroma and embedded in fibrous connective tissue. CONCLUSION: With optimal preparations a confident cytological diagnosis of endometriosis may be established easily, allowing correct treatment of the disease and, in selected cases, planning of preoperative pharmacologic therapy.

Thyroglossal duct cyst: case series.

OBJECTIVE: Thyroglossal duct cysts are remnants of the embryonic thyroglossal duct that may occur anywhere from the base of the tongue to the thyroid gland. The majority, however, are found at the level of the thyrohyoid membrane, under the deep cervical fascia. They are midline or just off the midline, and move up and down upon swallowing. This paper presents five case reports of TDC seen in the Department of Ear-Nose-Throat Surgery Regional Hospital Of Chania, Crete, Greece. It also discusses the different diagnostic approaches and differential diagnoses of the lesion. STUDY DESIGN: The medical records of patients admitted from 1995-2006 were reviewed for patients treated for TDC. History and examination reports were studied. When possible, results and reports of special investigations were obtained and the investigations were re-evaluated. Surgical operation notes and histology reports were obtained and the histologic slides were re-examined as necessary. RESULTS: Five cases of thyroglossal duct cyst treated in our department are described with each having a different clinical picture. CONCLUSIONS: Although the clinical and histological presentations of these five cases are not rare, they do illustrate how varied thyroglossal duct cysts can be with respect to patient age, anatomic site, or associated signs and symptoms.

The Avatar Acceptability Study: Survivor, Parent and Community Willingness to Use Patient-Derived Xenografts to Personalize Cancer Care.

BACKGROUND: Using patient-derived xenografts (PDXs) to assess chemosensitivity to anti-cancer agents in real-time may improve cancer care by enabling individualized clinical decision-making. However, it is unknown whether this new approach will be met with acceptance by patients, family and community. METHODS: We used a cross-sectional structured survey to investigate PDX acceptability with 1550 individuals across Australia and New Zealand (648 survivors of adult and childhood cancer, versus 650 community comparisons; and 48 parents of childhood cancer survivors versus 204 community parents). We identified factors influencing willingness-to-use PDXs, willingness-to-pay, maximum acceptable wait-time, and maximum acceptable number of mice used per patient. FINDINGS: PDXs were highly acceptable: >80% of those affected by cancer felt the potential advantages of PDXs outweighed the disadvantages (community participants: 68%). Survivors' and survivors' parents' most highly endorsed advantage was 'increased chance of survival'. 'Harm to animals' was the least endorsed disadvantage for all groups. Cancer survivors were more willing to use PDXs than community comparisons [p < ·001]. Survivors and survivors' parents were willing to pay more [p < ·001; p = ∙004 respectively], wait longer for results [p = ·03; p = ∙01], and use more mice [p = ·01; p < ∙001] than community comparisons. Male survivors found PDXs more acceptable [p = ·01] and were willing to pay more [p < ·001] than female survivors. Survivors with higher incomes found PDXs more acceptable [p = ·002] and were willing to pay more [p < ·001] than survivors with lower incomes. Mothers found PDXs more acceptable [p = ·04] but were less willing to wait [p = ·02] than fathers. INTERPRETATION: We found significant attitudinal support for PDX-guided cancer care. Willingness-to-pay and maximum acceptable number of mice align well with likely future usage. Maximum acceptable wait-times were lower than is currently achievable, highlighting an important area for future patient education until technology has caught up.

Effect of ternary phosphate-based glass compositions on osteoblast and osteoblast-like proliferation, differentiation and death in vitro.

There is currently a need to expand the range of graft materials available to orthopaedic surgeons. This study investigated the effect of ternary phosphate-based glass (PBG) compositions on the behaviour of osteoblast and osteoblast-like cells. PBGs of the formula (in mol.%) P(2)O(5)(50)-CaO(50-X)-Na(2)O(X), where X is either 2, 4, 6, 8 or 10, were produced and their influence on the proliferation, differentiation and death in vitro of adult human bone marrow stromal cells (hBMSCs) and human fetal osteoblast 1.19 (HFOB 1.19) cells were assessed. Tissue culture plastic (TCP) and hydroxyapatite (HA) were used as controls. Exposure to PBGs in culture inhibited cell adhesion and proliferation and increased cell death in both cell types studied. There was no significant difference in percentage cell death between the PBGs, which was significantly greater than the controls. However, compared with other PBGs, a greater number of cells were found on the 48mol.% CaO which may have been due to either increased adherence or proliferation, or both. This composition was capable of supporting osteogenic proliferation and early differentiation, and supports the notion that chemical modification of the glass could lead to a more biologically compatible substrate with the potential to support osteogenic grafting. Realisation of this potential should lead to the development of novel grafting strategies for the treatment of problematic bone defects.

Display of heterologous proteins on the surface of microorganisms: from the screening of combinatorial libraries to live recombinant vaccines.

In recent years there has been considerable progress towards the development of expression systems for the display of heterologous polypeptides and, to a lesser extent, oligosaccharides on the surface of bacteria or yeast. The availability of protein display vectors has in turn provided the impetus for a range of exciting technologies. Polypeptide libraries can be displayed in bacteria and screened by cell sorting techniques, thus simplifying the isolation of proteins with high affinity for ligands. Expression of antigens on the surface of nonvirulent microorganisms is an attractive approach to the development of high-efficacy recombinant live vaccines. Finally, cells displaying protein receptors or antibodies are of use for analytical applications and bioseparations.

Function-based isolation of novel enzymes from a large library.

Here we describe a high-throughput, quantitative method for the isolation of enzymes with novel substrate specificities from large libraries of protein variants. Protein variants are displayed on the surface of microorganisms and incubated with a synthetic substrate consisting of (1) a fluorescent dye (2) a positively charged moiety (3) the target scissile bond, and (4) a fluorescence resonance energy transfer (FRET) quenching partner. Enzymatic cleavage of the scissile bond results in release of the FRET quenching partner while the fluorescent product is retained on the cell surface, allowing isolation of catalytically active clones by fluorescence-activated cell sorting (FACS). Using a synthetic substrate with these characteristics, we enriched Escherichia coli expressing the serine protease OmpT from cells expressing an inactive OmpT variant by over 5,000-fold in a single round. Screening a library of 6 x 10(5) random OmpT variants by FACS using a FRET peptide substrate with a nonpreferred Arg-Val cleavage sequence resulted in the isolation of variant proteases with catalytic activities enhanced by as much as 60-fold. This approach represents a potentially widely applicable method for high-throughput screening of large libraries on the basis of catalytic turnover.