Strict Morphological Criteria Research Articles

Different Nuclear Architecture in Human Sperm According to Their Morphology.

Human sperm parameters serve as a first step in diagnosing male infertility, but not in determining the potential for successful pregnancy during assisted reproductive technologies (ARTs) procedures. Here, we investigated the relationship between sperm head morphology at high magnification, based on strict morphologic criteria, and the nuclear architecture analyzed by fluorescence in situ hybridization (FISH). We included five men. Two of them had an elevated high-magnification morphology score of 6 points (Score 6) indicating high fertility potential, whereas three had a low score of 0 points (Score 0), indicating low fertility potential. We used FISH to study the inter-telomeric distance and the chromosomal territory area of chromosome 1 (Chr. 1). We then compared these two parameters between subjects with high and low scores. FISH data analysis showed that the inter-telomeric distance (ITD) and chromosomal territory area (CTA) of Chr. 1 were significantly higher in subjects with low scores (score 0) than high scores (score 6). Our results suggest that (i) there is a link between nuclear architecture and sperm head abnormalities, particularly vacuoles; and (ii) it is possible to select spermatozoa with normal nuclear architecture, which might indirectly explain the positive ART outcomes observed with this technique.

Pulmonary Low Malignant Potential Adenocarcinoma: A Validation of the Proposed Criteria for This Novel Subtype.

Adenocarcinoma (ADC) is the most common histologic type of lung cancer, including in situ (lepidic), minimally invasive, and invasive forms. While the former 2 types are associated with a favorable outcome, the latter includes tumors with variable behavior, often tumor stage-related. A recent study proposed strict morphologic criteria defining a new subgroup of resected stage I invasive ADC (16% of cases) with favorable outcomes (100% disease-specific survival), named "ADC of low malignant potential (LMP-ADC)." The following criteria were met: ≤3cm size, nonmucinous histotype, ≥15% lepidic growth, and the absence of the following: high-grade patterns, >1 mitosis/2mm 2 , necrosis, and vascular/pleural invasion. The aim of the present study was to validate the performance of such criteria to identify LMP-ADC in a series of 274 stage IA resected lung ADCs from a single institution. Thirty-four tumors (12.4%) met the proposed criteria for LMP-ADC, as confirmed by additional stains for mitotic figures, Ki67 index, and elastic fibers (helpful to assess alveolar wall invasion). Minor differences between the lepidic and invasive components were observed regarding cell atypia and proliferation. p53 was normally expressed by invasive tumor cells. Mutations occurred in known lung cancer genes (mostly KRAS and EGFR). Five patients (14.7%) developed disease progression and 2 of them (5.9%) died of the disease. In our series, the disease-specific survival was 94.1%. In conclusion, in resected invasive lung ADC, a subgroup presenting low-grade morphologic features and associated with favorable prognosis does exist. Morphologic criteria for LMP-ADC supported by ancillary techniques represent a valid tool to better define this novel subgroup and to refine the stratification of invasive lung ADC, possibly suggesting modified follow-up protocols, based on the observed indolent behavior in most cases.

Left bundle branch block and ventricular dysfunction: the importance of QRS duration over strict morphological criteria; data from the RECOrd-LBBB registry

Abstract Background Left bundle branch block (LBBB) has been associated with left ventricular (LV) remodeling and dysfunction. LBBB may appear as an ominous sign in the natural course several heart diseases, or may play a causative role in the development of a “dyssynchronopathy” (LBBB-induced cardiomyopathy). LBBB represents an important therapeutic target in heart failure (HF) patients, since cardiac resynchronization therapy (CRT) aims to restore mechanical synchrony. Over the last 10 years different electrocardiographic (ECG) criteria (by the American Heart Association – AHA, and by Strauss) have been proposed to better identify and select patients suitable for CRT. The real usefulness of these criteria has been recently questioned. Purpose Aims of this single center registry were: 1) to describe the clinical and instrumental (ECG and echocardiographic) features in patients with LBBB hospitalized due to both cardiac and non-cardiac causes in a tertiary university hospital; 2) to identify the prevalence of LV remodeling/dysfunction, and its potential determinants. Methods One hundred eighty-six patients underwent clinical evaluation, 12-lead ECG, and transthoracic echocardiogram. Exclusion criteria were: 1) prior myocardial infarction with extensive anterior necrosis; 2) prior cardiac surgery or transcatheter aortic valve implantation; 3) complex congenital heart diseases; 4) permanent pacing with right ventricular stimulation. Results One hundred and three (55%) were males. Median age was 74.5 (IQR 66–82) years. Arterial hypertension was the most prevalent (65%) cardiovascular risk factor. Twenty-one patients had atrial fibrillation. Median QRS duration was 140 (IQR 140–160) msec, 70% of patients had a LBBB according to the AHA criteria, while 85% was diagnosed as having a “true” LBBB according to Strauss definition. A discordant LBBB was found in 56% of patients. Median LV ejection fraction (EF) was 42%, and LV systolic dysfunction was found in 66% of patients. There was a weak, although statistically significant, inverse correlation between QRS duration (msec) and LVEF (rho −0.23; p=0.0026). There was a significant difference in terms of QRS duration (160 vs 140 msec; p=0.008) between patients with and without LV dysfunction, while LBBB morphological criteria did not differentiate the two groups. QRS duration resulted to be the only independent determinant of LV systolic dysfunction (OR 1.022 [95% CI]=1.022 [1.002–1.042], p=0.029) on multivariable logistic regression analysis. Conclusions In a non-selected cohort of patients with LBBB, the use of strict morphological criteria does not predict the presence of LV dysfunction. QRS duration seems to be the only independent predictor of LV dysfunction. These results emphasize the complex relationship between LBBB and cardiac remodeling and dysfunction. Funding Acknowledgement Type of funding sources: None.

P-044 Abnormal sperm morphology, so what? The clinical outcome of medical assisted reproduction is not affected by 100% teratospermic semen samples

Abstract Study question What is the success rate (clinical outcome) in medical assisted reproduction (MAR) when 0% normal spermatozoa forms are observed using strict morphology criteria (SMC)? Summary answer Strict criteria morphology has no predictive value for the clinical outcome of MAR. Absence of normal morphology spermatozoa does not preclude high cumulative pregnancy rates. What is known already Many studies have questioned the clinical value of sperm morphology since no effect on treatment outcome is apparent when sperm samples with <4% normal forms are used. However, data evaluating treatment outcome when 0% normal sperm at diagnose (evaluated by strict morphology criteria, WHO 2010) scarce. While teratozoospermia values are known to have a negative correlation with fertilization per cycle, data on cumulative ongoing pregnancy rates (COPR) or live birth rates (LBR) are missing in this selected group. Study design, size, duration This is a retrospective analysis (January 2010-June 2021) of the clinical outcome for couples when the male partner presents with 0% normal morphology (WHO 2010) at diagnostic semen analysis. Cases with 100% globozoospermia, cilia abnormalities or other MMAF phenotypes were excluded. Depending on the total progressively motile sperm count (TPMSC), couples were referred for MAR. The primary outcome presented is COPR/treatment and LBR/couple. Secondary outcomes were total fertilization failure (TFF) and mean fertilization rate (FR). Participants/materials, setting, methods All 193 cases included in this study presented with 100% teratozoospermia, as evaluated by two independent technicians following WHO 2010 manual recommendations. Couples were treated with IUI,IVF or ICSI depending on TPMSC at diagnosis. A total of 17 couples underwent 68 IUI treatments, 20 couples underwent 36 IVF ovum-pickups (OPU) (with respectively fresh embryo transfer (ET) and frozen ET, FET); 156 couples underwent 394 ICSI-OPU (with respectively fresh-ET and FET). Cumulative-OPR per treatment/couple is presented. Main results and the role of chance From the 17 couples undergoing IUI (mean 4 IUI cycles/couple; with or without mild ovarian stimulation) 3 ongoing pregnancies were reported (all singletons; 18% LBR per couple). Twenty IVF couples (mean 1.8 OPUs/couple) resulted in a total of 13 ongoing pregnancies (12 singletons, 1 twin pregnancy), therefore, the LBR per IVF-couple was 65%. In six cycles a TFF was observed even with sufficient motile sperm during insemination (5x with TPMSC >5 x 10^6/ml). If TFF was observed after IVF cycle, then ICSI was performed in the next cycle. The IVF mean %FR using 100% teratozoospermic samples was 54%, compared to 58% in cases with >1% normal forms. Regarding ICSI, 156 couples underwent a total of 377 cycles (mean of 2.4 OPUs/couple). These treatments resulted in a total of 357 fresh-ET and 458 FET. A total of 103 couples achieved 154 ongoing pregnancies (146 singletons, 7 twins), therefore the COPR was 39%/cycle and LBR was 66%/couple. In 13 cycles (3.4%) TFF was observed and in 15 cases the fertilization rate was <20%. The mean FR with ICSI using teratozoospermic samples was 55%. Comparingly, using samples with >1% normal morphology, the laboratory mean for FR is 68% and 3.7% for TFF. Limitations, reasons for caution This is a retrospective study and not matched case-control cohort was used in this analysis. Not all couples have completed 3 complete IVF/ICSI cycles. As not all frozen embryos have been transferred, cumulative pregnancy rates might be higher than the rates reported here. Wider implications of the findings The data presented here confirms the limited value of the actual strict criteria for sperm morphology evaluation. Morphology is important for identification of sporadic sperm samples with atypical malformations such as globozoospermia, MMAF and other aberrant forms of a genetic origin, however SMC evaluation and interpretation should be reconsidered Trial registration number N.A.

Outcome reporting across randomized controlled trials evaluating potential treatments for male infertility: a systematic review.

STUDY QUESTIONWhat are the primary outcomes and outcome measures used in randomized controlled trials (RCTs) evaluating potential treatments for male infertility in the last 10 years?SUMMARY ANSWEROutcome reporting across male infertility trials is heterogeneous with numerous definitions and measures used to define similar outcomes.WHAT IS KNOWN ALREADYNo core outcome set for male infertility trials has been developed. Male infertility trials are unique in that they have potentially three participants, a man, a female partner and their offspring and this will likely lead to significant variation in outcome reporting in randomized trials.STUDY DESIGN, SIZE, DURATIONA systematic review of RCTs mapping outcomes and outcome measures evaluating potential treatments for men with infertility registered in the Cochrane Register of Controlled Trials (CENTRAL) between January 2010 and July 2021.PARTICIPANTS/MATERIALS, SETTING, METHODS screening and study selection was undertaken in duplicate using a review protocol that was developed prior to commencing the review. No risk of bias assessment was undertaken as this review aims to report on outcome reporting only.MAIN RESULTS AND THE ROLE OF CHANCEOne hundred and seventy-five RCTs were identified, and given the large number of studies we limited our review to the 100 largest trials. Seventy-nine different treatments were reported across the 100 largest RCTs including vitamin and dietary supplements (18 trials), surgical treatments (18 trials) and sperm selection techniques (22 trials). When considering the largest 100 trials (range: 80–2772 participants), 36 primary and 89 secondary outcomes were reported. Forty-seven trials reported a primary outcome and 36 trials clearly defined their primary outcome. Pregnancy outcomes were inconsistently reported and included pregnancy rate (51 trials), pregnancy loss including miscarriage, ectopic pregnancy, stillbirth (9 trials) and live birth (13 trials). Trials consistently reporting the same outcome frequently used different definitions. For example, semen quality was reported by 75 trials and was defined in 7 different ways, including; the World Health Organization (WHO) 2010 criteria (32 trials), WHO 1999 criteria (18 trials), WHO 1992 criteria (3 trials), WHO 1999 and 1992 criteria (1 trial) and the Kruger strict morphology criteria (1 trial).LIMITATIONS, REASONS FOR CAUTIONWe only evaluated the 100 largest trials published in the last 10 years and did not report outcomes on the remaining 75. An outcome was included as a primary outcome only if clearly stated in the manuscript and we did not contact authors to clarify this. As our review mapped outcomes and outcome measures, we did not undertake an integrity assessment of the trials included in our review.WIDER IMPLICATIONS OF THE FINDINGSMost randomized trials evaluating treatments for male infertility report different outcomes. Only half of the RCTs reported pregnancy rate and even fewer reported live birth; furthermore, the definitions of these outcomes varies across trials. Developing, disseminating and implementing a minimum data set, known as a core outcome set, for male infertility research could help to improve outcome selection, collection and reporting.STUDY FUNDING/COMPETING INTEREST(S)A.P.—chairman of external scientific advisory committee of Cryos International Denmark ApS, member of the scientific advisory board for Cytoswim LDT and ExSeed Health. Guest lecture at the ‘Insights for Fertility Conference’, funded by MERK SERONO Limited. M.v.W.—holds a ZON-MW research grant. No external funding was obtained for this study.

Assessing the clinical value of the Kruger strict morphology criteria over the World Health Organization fourth edition criteria

ObjectiveTo assess if the newer Kruger strict morphology (WHO5; normal ≥4%) adds any clinical value beyond the criteria of the World Health Organization fourth edition (WHO4; normal ≥14%).DesignRetrospective study.SettingTertiary hospital.Patient(s)Men without known azoospermia who had semen analysis (SA) collected over a 10-year period of time.Intervention(s)Morphology classification under Kruger WHO5 strict criteria and WHO4 criteria.Main Outcome Measure(s)Correlation between the WHO5 and WHO4 morphological classifications.Result(s)A total of 4,510 SAs were identified during the study period. Of these, both Kruger WHO5 and WHO4 morphologies were included in 932 SAs (20.7%) from a total of 691 men. The median age of the men was 37 years (interquartile range, 32.0–43.8 years). The mean (±SD) semen volume, sperm concentration, and motility were 2.6 ± 1.4 mL, 50.0 ± 35.6 × 106/mL, and 53.1% ± 18.6%, respectively. The correlation between the WHO4 and WHO5 morphology assessments was high (Spearman correlation coefficient = 0.94). Only 545 (58.5%) of 932 SAs had abnormal Kruger WHO5 morphology, of which 543 (99.6%) of 545 also had abnormal morphology by the WHO4 criteria.Conclusion(s)The Kruger WHO5 and WHO4 morphologic criteria correlate closely. Only two men (0.4%) with an abnormal Kruger morphology had normal WHO4 morphology. Given the limited predictive value of sperm morphology, the additional cost and effort of Kruger criteria may not be warranted in lieu of, or in addition to, the WHO4 classification.

Hepatocellular carcinoma

Hepatocellular carcinoma (HCC) belongs to the most prevalent and deadliest cancers worldwide. It can be attributed to well-defined risk factors (mainly chronic viral hepatitis and alcoholic/nonalcoholic steatohepatitis) leading to liver cirrhosis, apremalignant condition for the development of preneoplastic hepatocellular lesions and finally liver cancer. By applying strict morphological criteria and apanel of immunohistological markers, early HCC can be differentiated from its precursor lesions and other highly differentiated hepatocellular lesions even in most biopsy specimens. Integrative characterization led to the association of histological features and molecular subgroups of human HCC. This potentially relevant clinical development was recognized by the recently updated WHO classification of liver cancer resulting in the introduction of several HCC subtypes. These are characterized by adistinct combination of histological and molecular features, biological behavior, and clinical characteristics, allowing for adistinction from other HCC without specified features. Whether this development sets the corner stone for precision oncology of human HCC patients must be monitored.

Morphological and immunohistochemical approaches to the diagnosis of Hashimoto thyroiditis and mucosa associated lymphoid tissue lymphomas: An audit.

Most cases of autoimmune hypothyroidism are due to Hashimoto thyroiditis (HT). It is sometimes difficult to distinguish HT from mucosa associated lymphoid tissue (MALT) lymphoma using cytology or histology alone. This has necessitated the use of immunohistochemistry and other molecular techniques. A descriptive study was undertaken to determine the prevalence of MALT lymphoma and other associations of HT using histopathological and immunohistochemical methods. For testing of kappa and lambda antibodies, paraffin sections were prepared for immunohistochemical staining using a peroxidase-antiperoxidase immune complex method. Immunostaining in HT, demonstrated a polyclonal lymphoid population, as evidenced by dual positivity for kappa and lambda staining cells, whereas MALT lymphoma revealed a monoclonal lymphoid population, with strong positivity for kappa immunostaining and lack of lambda light chain expression. The proportion of MALT lymphoma in surgically treated cases of HT in the present study was 3.5% (95% CI: 0%–8.3%). The common malignancy associated with HT was papillary carcinoma of the thyroid (8.8%). Among the benign conditions, nodular colloid goiter (7%) was more commonly associated. Although modern molecular techniques are available for the confirmation of lymphomas, they are more expensive, time consuming and are available only in a few centers. Strict morphological criteria can differentiate HT from MALT lymphoma, but in suspicious cases, paraffin section immunohistochemistry using light chain restriction can offer comparable and reliable results. Since papillary carcinoma and MALT lymphoma have been associated with HT, these patients require careful surveillance.

Nonconventional dysplasia in patients with inflammatory bowel disease and colorectal carcinoma: a multicenter clinicopathologic study.

Several types of nonconventional dysplasia have been recently described in inflammatory bowel disease (IBD). However, strict morphologic criteria are lacking, and their clinicopathologic features (including potential association with conventional dysplasia and/or colorectal cancer [CRC]) are poorly understood. A total of 106 dysplastic or serrated lesions in 58 IBD patients with CRC were retrospectively identified from five institutions. Thirty-six cases of nonconventional dysplasia were identified in 26 (45%) of the 58 patients and occurred with similar frequency in men and women (58% and 42%, respectively), with a mean age of 54 years (range: 24-73) and a long history of IBD (mean: 17 years, range: 2-43). Six morphologic patterns were recognized. Hypermucinous dysplasia (n = 15; 42%) presented as either a 'pure type' (n = 5; 14%) or a 'mixed type' with either conventional or another nonconventional subtype (n = 10; 28%). Serrated lesions, as a group, were equally common (n = 15; 42%) and included three variants: traditional serrated adenoma-like (n = 10; 28%), sessile serrated lesion-like (n = 1; 3%), and serrated lesion, not otherwise specified (n = 4; 11%). Dysplastic lesions with increased Paneth cell differentiation (n = 4; 11%) and goblet cell deficient dysplasia (n = 2; 6%) were rare. Twelve (46%) of the 26 patients had only nonconventional dysplasia, whereas the remaining 14 patients (54%) had both nonconventional and conventional dysplasias. Nonconventional dysplasia was most often graded as low-grade dysplasia (81%), which was less common in conventional dysplasia (37%) (p = 0.003). When present alone, nonconventional dysplasia was predominantly found in the left colon (81%, p = 0.006) as a polypoid or raised lesion (75%, p < 0.001) compared with when it occurred simultaneously with conventional dysplasia (35% and 50%, respectively). When both nonconventional and conventional dysplasias occurred simultaneously, they were found in the same colonic segment in all but 3 patients (79%). Nonconventional dysplasia was also commonly detected in the same colonic segment as CRC or immediately adjacent to the CRC at a rate (85%) similar to conventional dysplasia (96%). CRC occurring in patients with only nonconventional dysplasia was more likely to be high-grade (poorly differentiated; 36%) than CRC that occurred in association with conventional dysplasia (10%) (p = 0.026). In conclusion, nonconventional dysplasia is common in IBD patients with CRC. It appears to develop in the same field of carcinomatous development, and it is not uncommonly associated with conventional dysplasia.

Tubulocystic renal cell carcinoma: a distinct clinicopathologic entity with a characteristic genomic profile

Tubulocystic renal cell carcinoma, a unique tumor, was recently included as a new entity in the World Health Organization classification of renal tumors. It has variably been reported to be related to other renal cell carcinomas, including papillary renal cell carcinoma, fumarate hydratase-deficient carcinoma, and others, likely because many such carcinomas may show variable amounts of tubulocystic architecture. The published data characterizing the molecular features of these tumors are inconsistent. We studied nine “pure” tubulocystic renal cell carcinomas, as defined by International Society of Urologic Pathologists (ISUP) and World Health Organization (WHO), by targeted next-generation sequencing, and fluorescence in situ hybridization for X and Y chromosomes, to investigate if these show any unique characteristics or any overlap with known mutational/molecular profiles or copy number alterations in other subtypes of renal cell carcinoma. All nine tubulocystic carcinomas demonstrated combined losses at chromosome 9 and gains at chromosome 17, as well as, loss of chromosome Y (in 5/5). None of the tumors showed mutational profiles characteristic of other renal neoplasms, including those seen in fumarate hydratase-deficient renal cell carcinoma. Recurrent mutations in chromatin-modifying genes, KMT2C and KDM5C, were detected in two of nine tumors. Thus, tubulocystic renal cell carcinoma, if defined strictly, at the clinical and pathologic level, demonstrates genomic features distinct from other subtypes of renal cell carcinoma. These findings support the contention that tubulocystic renal cell carcinoma should be diagnosed only using strict morphological criteria and only when presenting in a “pure” form; presence of variable papillary, poorly differentiated, or other architectural patterns most likely do not belong to the category of tubulocystic renal cell carcinoma.

Osteoblastoma‐like osteosarcoma: high‐grade or low‐grade osteosarcoma?

Osteoblastoma-like osteosarcoma is a rare variant of osteosarcoma (1% of all osteosarcomas), histologically similar to osteoblastoma. In the current World Health Organisation (WHO) classification, osteoblastoma-like osteosarcoma is classified within the group of conventional (high-grade) osteosarcomas. However, several published cases have been actually regarded as low-grade malignant tumours. As strict morphological criteria to distinguish between low- and high-grade lesions are not available, we reviewed our series of osteoblastoma-like osteosarcomas in the attempt to identify clinical and morphological features predictive of aggressiveness. We retrieved 15 cases of osteoblastoma-like osteosarcoma from the files of the Istituto Ortopedico Rizzoli. Patients received various treatments. Five patients developed metastasis and five patients developed local recurrences (all after incomplete surgery). Eleven patients were alive without disease, while four patients died of their disease. Statistical analysis revealed a statistically significant (P=0.048) lower disease-free survival in patients with areas of conventional (high-grade) osteosarcoma. With the important limitation of a small cohort of patients, the presence of areas of conventional (high-grade) osteosarcoma is the only parameter to predict the aggressiveness of osteoblastoma-like osteosarcoma.

"Non-invasive Follicular Thyroid Neoplasm with Papillary like Nuclear Features": An Institutional Retrospective Study with Re-nomenclature and Re-classification.

The cumulative efforts of pioneers in field of Thyroid pathology have resulted in proposed reclassification of Encapsulated Follicular Variant of Papillary Thyroid Carcinoma (EFVPTC) to "Non-invasive follicular thyroid neoplasm with papillary-like nuclear features" (NIFTWP) due to indolent clinical behaviour. To study the spectrum of EFVPTCs in our institute and to reclassify them as NIFTWP using stringent morphological criteria. This is a retrospective study conducted from Jan 2014 to July 2017. Institutional records were accessed and out of 485 thyroidectomy cases, 35 cases of EFVPTC were taken into study and field's kappa was used as a means of statistical analysis. The study conducted on 35 cases of EFVPTCs using strict morphological criteria lead to re-nomenclature and re-classification of 30 out of these 35 cases, with a field's kappa value of 4.3. The present study, lead to re-nomenclature and re-classification of 30 cases of EFVPTC to NIFTWP and proved statistically that there is moderate level of Inter-observer agreement regarding the criteria used for this re-nomenclature and re-classification.

An insect-like mushroom body in a crustacean brain.

Mushroom bodies are the iconic learning and memory centers of insects. No previously described crustacean possesses a mushroom body as defined by strict morphological criteria although crustacean centers called hemiellipsoid bodies, which serve functions in sensory integration, have been viewed as evolutionarily convergent with mushroom bodies. Here, using key identifiers to characterize neural arrangements, we demonstrate insect-like mushroom bodies in stomatopod crustaceans (mantis shrimps). More than any other crustacean taxon, mantis shrimps display sophisticated behaviors relating to predation, spatial memory, and visual recognition comparable to those of insects. However, neuroanatomy-based cladistics suggesting close phylogenetic proximity of insects and stomatopod crustaceans conflicts with genomic evidence showing hexapods closely related to simple crustaceans called remipedes. We discuss whether corresponding anatomical phenotypes described here reflect the cerebral morphology of a common ancestor of Pancrustacea or an extraordinary example of convergent evolution.

The genetic diversity, phylogeography and morphology of Elphidiidae (Foraminifera) in the Northeast Atlantic

Genetic characterisation (SSU rRNA genotyping) and Scanning Electron Microscope (SEM) imaging of individual tests were used in tandem to determine the modern species richness of the foraminiferal family Elphidiidae (Elphidium, Haynesina and related genera) across the Northeast Atlantic shelf biomes. Specimens were collected at 25 locations from the High Arctic to Iberia, and a total of 1013 individual specimens were successfully SEM imaged and genotyped. Phylogenetic analyses were carried out in combination with 28 other elphidiid sequences from GenBank and seventeen distinct elphidiid genetic types were identified within the sample set, seven being sequenced for the first time. Genetic types cluster into seven main clades which largely represent their general morphological character. Differences between genetic types at the genetic, morphological and biogeographic levels are indicative of species level distinction. Their biogeographic distributions, in combination with elphidiid SSU sequences from GenBank and high resolution images from the literature show that each of them exhibits species-specific rather than clade-specific biogeographies. Due to taxonomic uncertainty and divergent taxonomic concepts between schools, we believe that morphospecies names should not be placed onto molecular phylogenies unless both the morphology and genetic type have been linked to the formally named holotype, or equivalent. Based on strict morphological criteria, we advocate using only a three-stage approach to taxonomy for practical application in micropalaeontological studies. It comprises genotyping, the production of a formal morphological description of the SEM images associated with the genetic type and then the allocation of the most appropriate taxonomic name by comparison with the formal type description. Using this approach, we were able to apply taxonomic names to fifteen genetic types. One of the remaining two may be potentially cryptic, and one is undescribed in the literature. In general, the phylogeographic distribution is in agreement with our knowledge of the ecology and biogeographical distribution of the corresponding morphospecies, highlighting the generally robust taxonomic framework of the Elphidiidae in time and space.

Novel embryo selection techniques to increase embryo implantation in IVF attempts.

The final success of an IVF attempt depends on several steps and decisions taken during the ovarian stimulation, the oocyte retrieval, the embryo culture and the embryo transfer. The final selection of the embryos most likely to implant is the final step in this process and the responsibility of the lab. Apart from strict morphologic criteria that historically have been used in embryo selection, additional information on genetic, metabolomic and morphokinetic characteristics of the embryo is recently combined to morphology to select the embryo most likely to produce a pregnancy. In this manuscript, we review the most recent information on the current methods used for embryo selection presenting the predictive capability of each one. A literature search was performed on Pubmed, Medline and Cochrane Database of Systematic Reviews for published studies using appropriate key words and phrases with no limits placed on time. It seems that the combination of morphologic criteria in conjunction to embryo kinetics as documented by time-lapse technology provides the most reliable information on embryo quality. Blastocyst biopsy with subsequent comprehensive chromosome analysis allows the selection of the euploid embryos with the higher implantation potential. Embryo time-lapse imaging and blastocyst biopsy combined to comprehensive chromosome analysis are the most promising technologies to increase pregnancy rates and reduce the possibility of multiple pregnancies. However, further studies will demonstrate the capability of routinely using these technologies to significantly improve IVF outcomes.

Abstract P1-01-11: Nuclear immunohistochemical IKK-ϵexpression in flat epithelial atypia (FEA) of the breast: A predictor of ipsilateral ADH, in-situ or invasive malignancy?

Abstract Background:Flat Epithelial Atypia of the breast (FEA) is associated with in situ and invasive low grade neoplasia. However, the role of excision after FEA on biopsy is controversial as rates of upgrading to atypical ductal hyperplasia (ADH), ductal carcinoma in situ (DCIS) or invasive carcinoma in subsequent excision are relatively low. Problems include difficulties in inter-observer reproducibility and lack of morphologic or immunohistochemistry (IHC) tools that better identify cases at risk for concurrent ADH/Carcinoma. Nuclear image analysis may be useful, but is not widely available. IKK-ϵ, part of the NF-kB activating pathway, is absent in normal breast epithelium and non atypical (usual) ductal hyperplasia, but is over-expressed in &amp;gt;30% of breast cancers. In addition, in our experience ADH/DCIS shows IKK-ϵ staining, mostly cytoplasmic. Of note, in prostate cancer, nuclear accumulation of IKK-ϵ has been described in hormone sensitive prostate disease while cytoplasmic accumulation is associated with metastatic progression. No previous studies of IKK-ϵ levels in FEA are reported. Here we report IKK-ϵ status in FEA and correlation with ipsilateral, synchronous ADH, DCIS or invasive carcinoma. Method: Resection specimens from 61 patients with diagnosis of FEA were retrieved. Presence of ADH/carcinoma and laterality (ipsi or contralateral) was recorded. Synchronous neoplasia was defined as ADH, DCIS or invasive carcinoma diagnosed within 6 months of the diagnosis of FEA. Presence of FEA was confirmed by three observers using strict morphologic criteria. IHC for IKK-ϵ was performed using ABCAM, rabbit anti-IKK-ϵ (ab7891) and pH 6 citrate buffer heat-induced epitope retrieval for 20 minutes. IHC slides were scanned and FEA regions captured for blind scoring of nuclear and cytoplasmic staining. Cut off for positive nuclear staining was 10% and cytoplasmic staining was graded as negative, weak, moderate or strong positive. Results:40 patients had ipsilateral synchronous ADH/carcinoma, and 21 did not. Within these groups, 6 patients had contralateral ADH/carcinoma (2 with and 4 without ipsilateral neoplasia). While cytoplasmic staining showed no difference between the groups, nuclear positivity was more frequent in cases with ipsilateral synchronous ADH/carcinoma, χ2(1, N = 61) = 5.1, p = .025 (Table 1). In contrast, there was no correlation between IKK-ϵ staining and ADH/carcinoma in the opposite breast (p=.25). Table 1Nuclear IKK-eSynchronous Ipsilateral ADH/DCIS/Carcinoma Negative (%)Positive (%)TotalNegative10 (48)11 (52)21Positive10 (25)30 (75)40 Conclusion:Nuclear IKK-ϵ staining may prove useful in predicting synchronous ipsilateral ADH or malignancy in cases of FEA in biopsy material. Given its more frequent association with ipsilateral synchronous ADH/carcinoma, IKK-ϵ nuclear expression in FEA may represent a step in continuous local oncogenesis rather than a general marker of risk. Given the pleiotropic role of IKK-ϵ in growth and survival, the significance of the shift from nuclear staining in FEA to cytoplasmic staining in ADH/DCIS may reflect different signaling pathways and requires further investigation. Further validation of our findings in larger cohorts is necessary. Citation Format: Williams PA, Parra-Herran CE, Ayroud Y, Islam S, Gravel DH, Robertson SJ, Pratt C. Nuclear immunohistochemical IKK-ϵexpression in flat epithelial atypia (FEA) of the breast: A predictor of ipsilateral ADH, in-situ or invasive malignancy?. [abstract]. In: Proceedings of the Thirty-Eighth Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2015 Dec 8-12; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2016;76(4 Suppl):Abstract nr P1-01-11.

Morphological assessment of thin basement membrane disease.

Thin basement membrane disease is more common than IgA nephropathy or Alport syndrome, which are also associated with the presence of erythrocyturia. Very few reports on the disorder are available in the Polish literature. The objective of this work was to analyze the results from 83 patients with thin basement membrane syndrome as well as to formulate a proposal of strict morphological assessment criteria for the disorder. Attention was drawn to the requirement of thickness of the lamina densa rather than the entire basement membrane thickness and a sufficiently high number of loops featuring thinned lamina densa, namely at least 80% of loops, being taken into account. Occurrence of other morphological changes associated with the disorder and clinical symptoms other than erythrocyturia was also highlighted.

Influence of Sperm Morphology on the Outcome of Assisted Reproductive Technique—Intracytoplasmic Sperm Injection Cycles: A Retrospective Analysis

ABSTRACTAimThe aim of this study was to evaluate the influence of spermatozoa morphology on ICSI cycle outcome parameters in couples with male factor infertility.DesignRetrospective study.SettingUniversity-level tertiary care center.Patients and methodsOne hundred and forty-eight couples with male factor infertility who had undergone intracytoplasmic sperm injection (ICSI) cycle from 2010 to December 2012 were included in this analysis. The semen samples of the male partners were classified according to the three predictive categories of the Tygerberg strict criteria: excellent prognosis (&gt; 14% morphologically normal spermatozoa), good prognosis (5–14%) and poor prognosis (&lt; 5%).Main outcome measuresThe primary outcome was the embryo quality rate.ResultsPatients in the poor prognosis subgroups exhibited deficits in spermatozoa concentration, motility and total motile fraction. The variations in the outcome parameters of fertilization rate, embryo development rate and embryo quality did not correlate with sperm morphology.ConclusionOur study suggests that Kruger's strict morphology criteria of the fresh semen sample is not a good predictor for the ICSI cycle outcome.How to cite this articlePreetha RP, Kamath MS, Aleyamma TK, Muthukumar K. Influence of Sperm Morphology on the Outcome of Assisted Reproductive Technique-Intracytoplasmic Sperm Injection Cycles: A Retrospective Analysis. Int J Infertil Fetal Med 2015;6(3):122-127.

Can morphological variables be used to predict hip joint function? An examination of the anthropoid proximal femur and range of abduction (919.10)

Femoral morphology is often been used to make inferences about hip mobility in fossil anthropoids, abduction ability in particular. However, no study has shown that variation in hip morphology is actually related to hip mobility. If hip joint mobility is predicted by morphological variation, this would provide a new tool to more specifically evaluate locomotor adaptation in fossils.We present results of an in silico model of anthropoid hip joint mobility that has been validated on a large sample of extant taxa (hominoids, cercopithecids, Ateles, Cebus). Pelves and femora of the same taxa were laser scanned and polygonal models were digitally articulated. Maximum hip abduction was modeled using PolyWorks software using strict morphological criteria for joint movement. Several morphologies of the femur that are hypothesized to relate to hip mobility were quantified from the laser scan data using PolyWorks. Regression was used to assess the relationship between morphologies and functional abilities.Neck‐shaft angle and femoral head orientation were significantly related to degree of abduction. However, the majority of morphologies quantified were not significant predictors of hip abduction ability, suggesting that many of the morphologies attributed to high joint mobility may not relate to increased abduction ability. This work has applications to reconstructing locomotor abilities in fossil anthropoids.Grant Funding Source: Study supported by NSF (BCS 1232393, 0647557, 0716244), Wenner Gren Foundation, LSB Leakey Foundatio

Fibrosarcoma-like Lipomatous Neoplasm

The term "spindle cell liposarcoma" has been applied to liposarcomas (LPSs) composed predominantly or exclusively of spindled cells. These tumors have been considered variants of well-differentiated LPS (WDL), myxoid LPS, and spindle cell lipoma, suggesting that this is a heterogenous group of lesions. Using strict morphologic criteria and molecular and immunohistochemical analyses, we have identified a homogenous group of spindle cell lipomatous tumors, histologically and genetically distinct from other forms of LPS, which we have called "fibrosarcoma-like lipomatous neoplasm." Cases classified as "spindle cell LPS" or "low-grade LPS with spindle cell features" were reviewed. Final selection criteria included: (1) an exclusive low-grade spindle cell component resembling fibrosarcoma; (2) a mixture of bland fibroblastic cells resembling the preadipocyte and early-adipocyte stage of embryonic fat; and (3) molecular-genetic analysis that excluded other forms of lipomatous tumors. Of the initial 25 cases identified, comparative genomic hybridization (CGH) was uninformative in 2 cases; 5 were reclassified as WDL on the basis of molecular data (MDM2 amplification) and 6 as spindle cell lipoma (CGH profiles with a few gains and losses including a constant loss of chromosome 13 and frequent losses of chromosomes 16 and 6). The 12 remaining cases showed flat CGH profiles; of these cases, 11 were negative for DDIT3 gene rearrangements, and 1 result was uninterpretable. Patients ranged in age from 15 to 82 years (mean 50 y); male patients were affected slightly more often (7:5). Tumors arose in the deep (6) and superficial (3) soft tissue of the groin (4), buttock (3), thigh (2), flank (1), shoulder (1), and paratesticular tissue (1) and ranged in size from 2 to 20 cm (mean 7.5 cm). Clinical follow-up in 11 patients (9 mo to 20 y; mean 68 mo) showed no recurrences or metastases. As defined above, "fibrosarcoma-like lipomatous neoplasm" is a unique lipomatous tumor that should be distinguished from WDL/(low-grade) dedifferentiated LPS and myxoid LPS on combined histologic/molecular features because of its better prognosis.