Association of pro- and anti-inflammatory cytokines in preeclampsia.

INTRODUCTION: The pro- and anti-inflammatory cytokines play crucial role in the development and functions of placenta. Any changes in these cytokines may be associated with many pregnancy-related disorders like preeclampsia. Therefore, the present study is aimed to study the expression of pro-inflammatory (TNF-α, IL-6) and anti-inflammatory (IL-4, IL-10) cytokines in placenta and serum of preeclamptic pregnant women. MATERIAL AND METHODS: For this study, a total of 194 cases of preeclamptic and control cases were enrolled in two Groups as per the gestational age that is, Group I (28-36 weeks) and II (37 weeks onwards). The number of samples was 55 in Group I and 139 in Group II. The immunohistochemistry (IHC) and enzyme-linked immunosorbent assay (ELISA) were conducted on placenta and serum of both preeclamptic and normal samples, respectively. IHC results were revalidated by reverse transcriptase PCR (RT-PCR). RESULTS: Both Groups (I, II) of preeclampsia showed amended levels of pro- and anti-inflammatory cytokines in placental tissues and serum samples. The levels of TNF-α and IL-6 were significantly increased in preeclamptic cases (P = 0.0001, P = 0.0001) while the IL-4 and IL-10 were downregulated (P = 0.0001, P = 0.0001) in comparison to control. In addition, a negative correlation was also observed between the two in preeclampsia (P = 0.0001). CONCLUSION: The balanced ratio of pro- and anti-inflammatory cytokines is essential to regulate the maternal inflammation system throughout pregnancy. Therefore, the gradual cytokine profiling of the pregnant women may be useful for the management of preeclampsia.

HIF-1 Alpha and Placental Growth Factor in Pregnancies Complicated With Preeclampsia: A Qualitative and Quantitative Analysis.

BACKGROUND: The pathophysiology of preeclampsia is not clearly understood worldwide. Hypoxia inducible factor 1α (HIF-1α) is thought to be the preliminary factor for the hypoxic conditions prevailing in preeclampsia, which causes imbalance in the expression of angiogenic proteins. A proangiogenic protein, placental growth factor (PIGF), is reported to be dysregulated in preeclampsia. Therefore, this study focuses on the investigation of HIF-1α and PIGF in preeclamptic conditions and a possible molecular association between them. METHODS: Placental tissue (n = 45 + 45) and serum samples (n = 80 + 80) of preeclamptic patients and healthy control were collected and processed for the analysis of HIF-1α and PIGF by immunohistochemistry and enzyme-linked immunosorbent assay (ELISA). RESULTS: In preeclamptic group, the significant nuclear and cytoplasmic expression of HIF-1α was noticed in syncytiotrophoblast (P = 0.0001) but in control placenta, it was localized to cytoplasm (P = 0.0001). The intensity of PIGF expression was lower in syncytiotrophoblast cytoplasm (P = 0.0001) in preeclamptic cases as compared with control. Also, the significant upregulated concentration of HIF-1α and downregulated PIGF was observed in serum samples of preeclamptic woman (P = 0.0001). Thus, there was a significant direct negative correlation between HIF-1α and PIGF both at tissue and serum level (P < 0.01). CONCLUSION: The direct inverse association between HIF-1α and PIGF in serum and placental tissues may be responsible for the low oxidative stress and endothelial dysfunction, leading to the pathogenesis of preeclampsia.

Augmented Activity of Cyclooxygenase-2 in Tissue and Serum of Patients With Cervical Cancer.

BACKGROUND: Cyclooxygenase-2 (Cox-2) is frequently overexpressed in cervical carcinoma, but little is known about its altered serum concentration. Hence, this study evaluates clinical utility of cellular and serum level of Cox-2 enzyme in cervical cancer. METHODS: The expression of Cox-2 was evaluated in cervical tissues and serum samples collected from normal controls (n = 100; n = 68), cervical intraepithelial neoplasia patients (CIN, n = 67; n = 12), and invasive squamous cell carcinoma patients (SCCs; n = 153; n = 127) by immunohistochemical and enzyme-linked immunosorbent assay (ELISA) analyses. RESULTS: The significant cytoplasmic overexpression of Cox-2 was noted in 50.7% of CIN and 69.9% of SCCs as compared with normal (P = 0.0001). Serum level of Cox-2 was also found to be elevated both in CIN (median 4.35 ng/ml) and in SCCs (median 19.39 ng/ml) with respect to normal (median 0.44 ng/ml; P = 0.0001), respectively. The ROC analysis revealed the potential of serum Cox-2 over its cellular expression to distinguish CIN and SCCs from normal. CONCLUSION: Augmented Cox-2 activity is implicated in the pathogenesis of cervical cancer, and its serum level could serve a potential to distinguish this malignancy. Therefore, it is suggested that serum Cox-2 may be useful in monitoring the diagnosis and treatment outcome of patients.

Aberrant pancreatic ductal organisation: a case report.

Anomalous pancreatic ductal system has always enthralled the anatomists, surgeons, gastroenterologists and radiologists alike. With the growing use of MRCP, ERCP and endoscopic and surgical procedures, the knowledge of anatomical aberrations of pancreaticobiliary tract becomes extremely important. Moreover, the anomalous pancreatic duct morphology may be responsible for atypical gastrointestinal complains. We report an exceptionally rare case of two accessory ventral pancreatic ducts opening separately into the common bile duct proximal to the hepato pancreatic ampulla. Concomitant occurrence of an ectopic major duodenal papilla, 3.8 cm distal to the pyloric end of stomach was also seen. Moreover, the accessory pancreatic duct and the minor duodenal papilla were absent. Clinical implications and embryological description of these rare anomalies are discussed. Awareness of such rare morphological variants can go a long way in assisting effective patient management.

Amalgamation of Toll-Like Receptor and Hypoxic Signaling in Etiology of Preeclampsia.

Inflammation and oxidative stress are involved in the pathogenesis of preeclampsia. Therefore, the aim of this study was to investigate the expression of Toll-like receptor (TLR) (TLR-4, HMGB1, NFκB, IκBα) and hypoxic (HIF-1α, HIF-1ß, PHD, pVHL) pathway proteins in the placenta of preeclamptic pregnant women after 28 weeks of gestational period. A possible association between these 2 pathways was also explored. A total of 194 placental tissues of preeclamptic as well as healthy pregnant women were analyzed by immunohistochemistry. On the basis of gestational age, the samples were divided into 2 groups, I (28-36 wk) and II (36 wk onwards), with 55 and 139 samples in the respective groups. The expression of both TLR (TLR-4, HMGB1, NFκB, IκBα) and hypoxic (HIF-1α, HIF-1ß, PHD, pVHL) pathway proteins were significantly modulated in the placental tissues of preeclampsia as compared with control. The 2 pathways were interlinked in preeclampsia. This study highlights the intercorrelation of both TLR and hypoxic signalling pathways that may be a causative factor for the pathophysiology of preeclampsia.

Crucial neurovascular structures entrapped in a brachial intramuscular tunnel.

We report a case where the median nerve accompanied by brachial vessels were found to traverse an intra-muscular tunnel within the brachialis muscle, in the floor of the cubital fossa. The muscular tunnel commenced 5 cm proximal to the neck of radius, measured 4.4 cm in length, and was present unilaterally. This unusual tunnel was distally found to blend with the brachial fascia. The present study was planned with an endeavor to elucidate in an improved way the clinical implications of compressed median nerve and brachial vessels. There are several sites where median nerve maybe compressed along its course in the arm and forearm. The relevance of the present documentation lies in the fact that these vital neurovascular structures may be compressed leading to neuropathies and vascular changes.

Rectus thoracis bifurcalis: a new variant in the anterior chest wall musculature.

A cadaveric study was undertaken to report the incidence of sternalis muscle in cadavers of Asian origin. A total of 88 cadavers were studied over a period of six years and the sternalis was reported only in a single case and that too unilaterally. The accessory muscle was discovered in the right pectoral region in a 40-year-old male cadaver. The muscle emanated from the external oblique aponeurosis of abdomen confirming its origin from the ventral longitudinal sheet of muscle. The muscle was fleshy throughout its extent except at the ends where they were aponeurotic. At the sternal angle, the muscle displayed "Y" shaped configuration and merged with the respective sternocleidomastoid muscle. The innervation was derived from the third intercostals nerve. We intend to highlight a few points through this study. Firstly, we found a paucity of studies undertaken to describe the incidence of sternalis muscle. Further, the studies present in anatomical archives are mainly case reports. Secondly, this muscle presents itself in varying configurations on radiological studies. The radiologist should acquaint himself with all these presentations, so that he can make accurate diagnosis of a breast mass. Thirdly, this muscle having more morphological relevance may be conveniently utilized for flap procedures of post mastectomy breast reconstruction. Lastly, the presence of this muscle may alter the depth at which the internal mammary lymph nodes are irradiated in case of carcinoma breast. Additionally, it should not be erroneously diagnosed as a mass which recurred on follow up of breast cancer patients. The present investigation endeavors to discuss the anatomical, embryological and clinical relevance of a rare accessory muscle of the anterior chest wall.

Anomalous superior thyroid artery.

The variations in the pattern of distribution of superior thyroid artery assume paramount importance for neck surgeons, in view of its vital topographical relationship to the external laryngeal nerve. In this study, we report an unusual variation in the arterial supply of the thyroid gland, which was detected during a routine dissection of an adult male cadaver. The right superior thyroid artery was absent whereas the left superior thyroid artery took origin from the left common carotid artery and showed a dominant pattern of distribution supplying the superior aspect of both the left and right lobes of the thyroid gland. It exhibited a usual relationship with the left external laryngeal nerve. The inferior thyroid arteries did not show any unusual distribution. Knowledge of such arterial variations related to the thyroid gland is immensely helpful for surgeons in order to put ligature on anomalous artery and to avoid damage to vital structures in this area, such as the external laryngeal nerve.

Author Correction: Cyclin D1 protein affecting global women's health by regulating HPV mediated adenocarcinoma of the uterine cervix.

An amendment to this paper has been published and can be accessed via a link at the top of the paper.

Expression analysis of E-cadherin, Slug and GSK3beta in invasive ductal carcinoma of breast.

BACKGROUND: Cancer progression is linked to a partially dedifferentiated epithelial cell phenotype. The signaling pathways Wnt, Hedgehog, TGF-beta and Notch have been implicated in experimental and developmental epithelial mesenchymal transition (EMT). Recent findings from our laboratory confirm that active Wnt/beta-catenin signaling is critically involved in invasive ductal carcinomas (IDCs) of breast. METHODS: In the current study, we analyzed the expression patterns and relationships between the key Wnt/beta-catenin signaling components- E-cadherin, Slug and GSK3beta in IDCs of breast. RESULTS: Of the 98 IDCs analyzed, 53 (54%) showed loss/or reduced membranous staining of E-cadherin in tumor cells. Nuclear accumulation of Slug was observed in 33 (34%) IDCs examined. Loss or reduced level of cytoplasmic GSK3beta expression was observed in 52/98 (53%) cases; while 34/98 (35%) tumors showed nuclear accumulation of GSK3beta. Statistical analysis revealed associations of nuclear Slug expression with loss of membranous E-cadherin (p = 0.001); nuclear beta-catenin (p = 0.001), and cytoplasmic beta-catenin (p = 0.005), suggesting Slug mediated E-cadherin suppression via the activation of Wnt/beta-catenin signaling pathway in IDCs. Our study also demonstrated significant correlation between GSK3beta nuclear localization and tumor grade (p = 0.02), suggesting its association with tumor progression. CONCLUSION: The present study for the first time provided the clinical evidence in support of Wnt/beta-catenin signaling upregulation in IDCs and key components of this pathway - E-cadherin, Slug and GSK3beta with beta-catenin in implementing EMT in these cells.

An Assembly of Anomalous Extensor Tendons of the Hand - Anatomical Description and Clinical Relevance.

Innumerable descriptions about variations in the pattern of extensor tendons are recorded in the literature. The dorsum of the hand in an adult male cadaver revealed an unusual pattern of extensor arrangement during a gross anatomical practical session. The extensor digitorum, extensor indicis and extensor digiti minimi tendons displayed a variant pattern. Extensor digitorum contributed tendons only to the middle and ring fingers, with junctura tendinum present between the extensor digitorum for the ring finger and extensor digiti minimi. Interestingly, an accessory muscle was observed arising from the common extensor origin passing to the index finger, in addition to the usual extensor indicis. The origin and insertion of extensor digiti minimi was as usual with an accessory slip contributed from the extensor carpi ulnaris to the proximal phalanx of the fifth finger. The plethora of variations in this region is of paramount importance for the reconstructive surgeon, who may utilize the accessory tendons to restore functional capacity of the fingers.

An assembly of anomalous extensor tendons of the hand--anatomical description and clinical relevance.

Innumerable descriptions about variations in the pattern of extensor tendons are recorded in the literature. The dorsum of the hand in an adult male cadaver revealed an unusual pattern of extensor arrangement during a gross anatomical practical session. The extensor digitorum, extensor indicis and extensor digiti minimi tendons displayed a variant pattern. Extensor digitorum contributed tendons only to the middle and ring fingers, with junctura tendinum present between the extensor digitorum for the ring finger and extensor digiti minimi. Interestingly, an accessory muscle was observed arising from the common extensor origin passing to the index finger, in addition to the usual extensor indicis. The origin and insertion of extensor digiti minimi was as usual with an accessory slip contributed from the extensor carpi ulnaris to the proximal phalanx of the fifth finger. The plethora of variations in this region is of paramount importance for the reconstructive surgeon, who may utilize the accessory tendons to restore functional capacity of the fingers.

Unilateral partial ossification of sacrotuberous ligament: anatomico-radiological evaluation and clinical implications.

The present study describes the topography and morphometry of a unilateral ossified sacrotuberous ligament. It aims to discuss its anatomical, radiological and clinical implications. The pudendal nerve, internal pudendal artery, nerve to obturator internus and coccygeal branch of inferior gluteal artery are all-important structures near sacrotuberous ligament. An ossified sacrotuberous ligament may be an important etiological factor in neurovascular compression syndromes and its anatomical knowledge may help in the development of new treatment for this common clinical problem. The ossified sacrotuberous ligament in the present case was 7.6 cm in length and exhibited a characteristic anterior and posterior segment. The base was at the ischial tuberosity and the apex showed numerous small bony protuberances with deep intervening grooves. The ossified STL may be important in differential diagnosis of soft tissue pain and tenderness after trauma. It may be a challenging puzzle for the present day surgeon and radiologist in interpretation of CT-scans and MRI.

Peculiar tendinous origin of the brachialis muscle: anatomic and clinical insight.

The current study seeks to elucidate the presence and potential function of an anomalous tendinous origin of the brachialis muscle. The brachialis muscle originates from the humeral shaft, which is usually muscular, thereby becoming tendo-aponeurotic distally towards its insertion. During routine cadaveric dissection, in a 52-year-old cadaver, the anomalous brachialis muscle originated in the form of a tendon measuring 4.4 cm in length. The tendon was attached to the proximal humeral shaft near the lateral lip of inter-tubercular sulcus. The aponeurotic insertion was as usual, into the ulnar coronoid process. The long tendinous origin of the brachialis muscle as observed in the present study could alter the biomechanics of the muscle. A long tendinous origin of the muscle may predispose to frequent tendinous injuries. There are possibilities of the anomalous tendon being used as a graft in elbow injuries. A comprehensive knowledge of such a rare anatomical variation could prove beneficial in surgical, rehabilitation and radiological field.

≤ Cyclin D1 protein affecting global women's health by regulating HPV mediated adenocarcinoma of the uterine cervix.

Adenocarcinoma (ADC) of the uterine cervix (UC) is a rare form of cervical cancer (CC) caused due to the infection of Human Papilloma Virus (HPV). Cyclin D1 is one of the downstream targets of aberrantly activated Notch signaling, contribute to the etiology of CC. However, little is known about the role of Cyclin D1 in the modulation of cervical ADC and is controversial. The purpose of this study is to determine the role of Cyclin D1 protein and to elucidate the combined analysis with Notch signaling proteins in HPV associated ADCs of CC. A total of 60 biopsy samples (40 normal and 20 ADCs of CC) were analyzed for the expression of Cyclin D1 in HPV associated ADCs via immunohistochemistry and by immunoblotting. HPV-16 positive ADC patients showed a strong association with the Cyclin D1 expression (p = 0.007). The significant mean difference (p = 0.0001) and the pairwise comparison between Cyclin D1/JAG1 (p = 0.0001), and Cyclin D1/Notch-3 (p = 0.0001) were observed. The above Notch signaling proteins showed their synergistic role in modulating Cyclin D1 which in-turn regulates HPV-16 associated ADC of the uterine cervix (UC), affecting women's global health.

HES1 Protein Modulates Human Papillomavirus-Mediated Carcinoma of the Uterine Cervix.

PURPOSE: Cervical cancer (CC) is the most common cancer affecting women worldwide. Human papillomavirus (HPV) infection is a major contributing factor for the development of CC. The development of CC occurs progressively from precancer stages to cancerous stages (ie, invasive squamous cell carcinoma [ISCC] and adenocarcinoma [ADC]). ADC is a rare form of CC that develops from the mucinous endocervical epithelium. It is believed that the downstream targets of Notch signaling contribute to the etiology of CC. One such target is HES1, whose role in the modulation of ADC is unknown. The purpose of this study is to determine the role of HES1 protein in HPV-associated ADC subtype of CC and also to compare its expression in histologic subtypes of precancer and ISCC. PATIENTS AND METHODS: A total of 148 patients (30 with precancers, 98 with ISCC, and 20 with ADC) and 40 normal control participants were analyzed for the expression of HES1 via immunohistochemistry, with results validated by immunoblotting. RESULTS: The comparison between HPV-16 and HES1 expression was significant in precancer (cervical intraepithelial neoplasia grades 1 to 3; P = .013), ISCC (International Federation of Gynecology and Obstetrics stages I to IV; P = .001), and ADC ( P = .007). An overall significant mean difference was observed between HES1, JAG1, and Notch-3 proteins in precancer ( P = .001), ISCC ( P = .001), and ADC ( P = .001). Pairwise comparisons between HES1 and JAG1 and HES1 and Notch-3 were also found to be significant. CONCLUSION: This study showed that among all HPV-16-positive precancers, the major HES1 positivity signal arises from cervical intraepithelial neoplasia grades 2 and 3 that develops into ISCC. Moreover, HPV-16-positive ADC also showed an association with HES1. The HES1, JAG1, and Notch-3 proteins showed their synergistic role in modulating HPV associated ADC along with histologic subtypes of precancer and ISCC of CC.

Anomalous pectoral musculature.

Anomalous disposition of pectoral muscles was encountered in an adult female cadaver on the left side. A prominent cleft separating the sternocostal and clavicular portions of the pectoralis major was noticed. The fibers of pectoralis major were partially fused with the deltoid, resulting in obliteration of the deltopectoral groove. Interestingly, cephalic vein was seen traversing superficial to the clavicular portion of the pectoralis major and pierced it to drain into the axillary vein. The pectoralis minor was inserted mainly on the coracoid process and few fibers were found blending with the coracobrachialis and short head of biceps brachii. Further, pectoralis minimus, a rare anatomic variant, was also observed lying superior to pectoralis minor. It was innervated by a twig from the lateral pectoral nerve at its superficial surface. Awareness of possibility of such anomalous muscles is important for surgeons operating on the chest wall.

Epigenetic alterations of CDH1 and APC genes: relationship with activation of Wnt/beta-catenin pathway in invasive ductal carcinoma of breast.

Activation of canonical Wnt/beta-catenin pathway in Invasive Ductal Carcinoma of Breast (IDCs) was recently reported from our laboratory. Herein, we analyzed promoter methylation status of CDH1 and Adenomatous polyposis coli (APC) genes in 50 IDCs and correlated with expression of E-cadherin (E-CD) and APC proteins and with activation of oncogenic Wnt/beta-catenin signaling pathway components, Dvl, beta-catenin and CyclinD1. Further, Wnt/beta-catenin driven epithelial mesenchymal transition (EMT) was investigated by correlating the expression of Dvl, beta-catenin and CyclinD1 with vimentin expression in these IDCs. Promoter hypermethylation was observed in 25/50 (50%) IDCs for CDH1 and in 11/50 (22%) tumors for APC, associated with loss of expression of E-CD and APC proteins; concordant hypermethylation of these genes was observed in paired patients' sera. Further, 57% of tumors harboring CDH1 methylation and 50% tumors harboring the methylated APC gene showed nuclear localization of beta-catenin, suggesting activation of the canonical Wnt/beta-catenin pathway. Our study demonstrates significant association between vimentin expression and nuclear beta-catenin (p=0.001; Odds ratio (OR)=25.6) and Dvl (p=0.023; OR=8.0), suggesting that EMT may be driven by Wnt/beta-catenin activation in IDCs. In conclusion, we demonstrate correlation of CDH1 and APC promoter methylation with loss of E-CD and APC proteins and with activation of Wnt/beta-catenin signaling pathway. Association of nuclear Dvl and beta-catenin with vimentin expression suggests the importance of Wnt/beta-catenin pathway driven EMT in IDCs. The concordance between CDH1 and APC methylation in IDCs and paired circulating DNA underscores the utility of serum DNA as a non-invasive tool for methylation analysis in IDC patients.

An anatomical insight into the third head of biceps brachii muscle.

BACKGROUND: The biceps brachii muscle is known to show variations in the number of heads. This study was performed to evaluate the variations in the origin of the biceps brachii muscle heads. MATERIALS AND METHODS: Both extremities of 48 formalin fixed cadavers (n = 96) were studied for abnormal heads of biceps brachii muscle. The flexor compartment of the arm was dissected and the attachments of the biceps brachii muscle were studied in detail. Appropriate photographs were taken. RESULTS: Among ninety six upper limbs studied, we observed three heads of biceps on both sides of a male cadaver aged 56 yrs (3.33%). The two heads of the biceps arose from its usual position but the anomalous third head arose from the anterior limb of the 'V' shaped insertion of the deltoid muscle on the humerus. The third head was found to fuse with the common belly of the muscle well before the bicipital tendon and its aponeurosis. Bilaterally this third head was supplied by a twig of the musculocutaneous nerve. No other abnormalities relating to the biceps were observed in any of the sides of the other 47 cadavers (n = 94). CONCLUSION: Our study revealed that the incidence of third head of biceps brachii may be approximately 3.33 % but larger studies are needed to confirm this fact. The third head of biceps brachii may be an incidental finding at autopsy or during routine anatomical dissections. Unless symptomatic, the third head of biceps brachii may not be detected in clinical studies (Fig. 3, Ref. 15).

Jagged-1 induced molecular alterations in HPV associated invasive squamous cell and adenocarcinoma of the human uterine cervix.

The majority of cervical cancer (CC) cases are attributable to HPV infection. Altered Notch pathway signals and HPV are believed to modify clinicopathogenesis of CC, however, the involvement of each molecular player and its mechanism is still not known. Jagged-1 (JAG1) is one of the ligands that induce Notch pathway. The involvement of JAG1 in the modulation of a disease condition is not very clear. Hence, this study aims to study the role of JAG1 in HPV-16/18 associated different histological sub-types of CC, especially ADC. 40 non-neoplastic cervical tissues, 30 precancer and 118 tumor specimens (total 188 tissue biopsies) were studied for the expression of the JAG1 protein through immunohistochemistry, immunoblotting and for HPV infection. Two folds increase of cytoplasmic (Mean ± S.E, 3.67 ± 0.33; p = 0.0001) and nuclear (3.70 ± 0.38, p = 0.0001) JAG1 expression was identified in normal (N) vs precancer and three folds cytoplasmic (4.44 ± 0.17, p = 0.0001) and nuclear (4.64 ± 0.17; p = 0.0001) in N vs. ISCC. Total 85% of ADC patients were found to be infected with HPV, which were 100% infected with HPV-16. These findings suggest the complex synergistic interplay between JAG1 and HPV in regulating clinicopathological progression of CC through its deregulation.