Rotenone-induced apoptosis and role of calcium: a study on Neuro-2a cells

Rotenone causes cytotoxicity in astrocytic cell culture by glial activation, which is linked to free radical generation. The present study is an investigation to explore whether rotenone could also cause cellular toxicity in mouse neuroblastoma cells (Neuro-2a) under treatment similar to astroglial cells. The effect of rotenone (0.1, 1, and 10?μM) on mitochondrial dehydrogenase enzyme activity by MTT reduction assay, PI uptake, total reactive oxygen species (ROS)/superoxide levels, nitrite levels, extent of DNA damage (by comet assay), and nuclear morphological alteration by Hoechst staining was studied. Caspase-3 and Ca(2+)/calmodulin-dependent protein kinase II (CaMKIIα) gene expression was determined to evaluate the apoptotic cell death and calcium kinase, respectively. Calcium level was estimated fluorometrically using fura-2A stain. Rotenone decreased mitochondrial dehydrogenase enzyme activity and generated ROS, superoxide, and nitrite. Rotenone treatment impaired cell intactness and nuclear morphology as depicted by PI uptake and chromosomal condensation of Neuro-2a cells, respectively. In addition, rotenone resulted in increased intracellular Ca(+2) level, caspase-3, and CaMKIIα expression. Furthermore, co-exposure of melatonin (300?μM), an antioxidant to cell culture, significantly suppressed the rotenone-induced decreased mitochondrial dehydrogenase enzyme activity, elevated ROS and RNS. However, melatonin was found ineffective to counteract rotenone-induced increased PI uptake, altered morphological changes, DNA damage, elevated Ca(+2), and increased expression of caspase-3 and CaMKIIα. The study indicates that intracellular calcium rather than oxidative stress is a major factor for rotenone-induced apoptosis in neuronal cells.     

A growing global network's role in outbreak response: AFHSC-GEIS 2008-2009

A cornerstone of effective disease surveillance programs comprises the early identification of infectious threats and the subsequent rapid response to prevent further spread. Effectively identifying, tracking and responding to these threats is often difficult and requires international cooperation due to the rapidity with which diseases cross national borders and spread throughout the global community as a result of travel and migration by humans and animals. From Oct.1, 2008 to Sept. 30, 2009, the United States Department of Defense's (DoD) Armed Forces Health Surveillance Center Global Emerging Infections Surveillance and Response System (AFHSC-GEIS) identified 76 outbreaks in 53 countries. Emerging infectious disease outbreaks were identified by the global network and included a wide spectrum of support activities in collaboration with host country partners, several of which were in direct support of the World Health Organization's (WHO) International Health Regulations (IHR) (2005). The network also supported military forces around the world affected by the novel influenza A/H1N1 pandemic of 2009. With IHR (2005) as the guiding framework for action, the AFHSC-GEIS network of international partners and overseas research laboratories continues to develop into a far-reaching system for identifying, analyzing and responding to emerging disease threats.     

Maternal and child health in Yushu, Qinghai Province, China

Introduction

Surmang, Qinghai Province is a rural nomadic Tibetan region in western China recently devastated by the 2010 Yushu earthquake; little information is available on access and coverage of maternal and child health services.

Methods

A cross-sectional household survey was conducted in August 2004. 402 women of reproductive age (15-50) were interviewed regarding their pregnancy history, access to and utilization of health care, and infant and child health care practices.

Results

Women's access to education was low at 15% for any formal schooling; adult female literacy was <20%. One third of women received any antenatal care during their last pregnancy. Institutional delivery and skilled birth attendance were <1%, and there were no reported cesarean deliveries. Birth was commonly attended by a female relative, and 8% of women delivered alone. Use of unsterilized instrument to cut the umbilical cord was nearly universal (94%), while coverage for tetanus toxoid immunization was only 14%. Traditional Tibetan healers were frequently sought for problems during pregnancy (70%), the postpartum period (87%), and for childhood illnesses (74%). Western medicine (61%) was preferred over Tibetan medicine (9%) for preventive antenatal care. The average time to reach a health facility was 4.3 hours. Postpartum infectious morbidity appeared to be high, but only 3% of women with postpartum problems received western medical care. 64% of recently pregnant women reported that they were very worried about dying in childbirth. The community reported 3 maternal deaths and 103 live births in the 19 months prior to the survey.

Conclusions

While China is on track to achieve national Millennium Development Goal targets for maternal and child health, women and children in Surmang suffer from substantial health inequities in access to antenatal, skilled birth and postpartum care. Institutional delivery, skilled attendance and cesarean delivery are virtually inaccessible, and consequently maternal and infant morbidity and mortality are likely high. Urgent action is needed to improve access to maternal, neonatal and child health care in these marginalized populations. The reconstruction after the recent earthquake provides a unique opportunity to link this population with the health system.     

Salmonella Berta myocarditis: Case report and systematic review of non-typhoid Salmonella myocarditis

AIM: To study trends in the epidemiology, clinical presentation, microbiology and prognosis of non-typhoid Salmonella (NTS) myocarditis.METHODS: We performed a systematic literature search for all reported NTS cases. The search yielded 838 publications. A total of 21 papers were deemed eligible. No language restrictions were enforced. Articles that were not written in English were translated. Pre-specified data such as clinical presentation, electrocardiogram (ECG) changes, transthoracic echocardiographic findings, cardiac magnetic resonance findings, microbiology cultures, Salmonella species, inflammatory markers (erythrocyte sedimentation rate and C-reactive protein), cardiac biomarkers and severity of illness were collected using data extraction sheets. Cases were classified by age into 2 groups; pediatric cases (defined as < 18 years old) and adult cases (defined ≥ 18 years old). The mean age of patients and standard deviations were calculated. The data was analyzed with IBM SPSS Statistics (Windows, Version 20.0. Armonk, NY: IBM Corp.) for demographic characteristics, presenting symptoms, microbiology, diagnostic methods, treatment modalities and outcome.RESULTS: From the selected articles, we identified a total of 24 individual cases with verifiable data. There were 20 males with a male to female ratio of 5:1. The mean age at presentation was 30.8 years (range 1 mo-67 years), 16% of cases were children aged < 18 years. Most patients presented with chest pain, fever, and abdominal pain. The most common ECG finding was ST elevation. Cardiac biomarkers were elevated in around 70% of cases. Salmonella Enteritidis was the most common NTS isolated. Definitive diagnosis was established by blood and stool cultures in most of the cases. The pediatric and adults cases had similar incidence of bacteremia (40% vs 36.8%) while the pediatric group had more stool cultures positive compared to the adult group (100% vs 63.1%). Eighty-three percent of patients received antibiotics and 58% were successfully treated through conservative management. The overall mortality was 24% and 42% of patients required intensive care.CONCLUSION: This systematic review of published cases shows that NTS myocarditis occurs predominantly in young adults and carries a poor prognosis.     

Metabolic Enhancer Piracetam Attenuates the Translocation of Mitochondrion-Specific Proteins of Caspase-Independent Pathway,Poly [ADP-Ribose] Polymerase 1 Up-regulation and Oxidative DNA Fragmentation

Piracetam, a nootropic drug, has been clinically used for decades; however, its mechanism of action still remains enigmatic. The present study was undertaken to evaluate the role of mitochondrion-specific factors of caspase-independent pathway like apoptotic-inducing factor (AIF) and endonuclease-G (endo-G) in piracetam-induced neuroprotection. N2A cells treated with lipopolysaccharide (LPS) exhibited significant cytotoxicity, impaired mitochondrial activity, and reactive oxygen species generation which was significantly attenuated with piracetam co-treatment. Cells co-treated with LPS and piracetam exhibited significant uptake of piracetam in comparison to only piracetam-treated cells as estimated by liquid chromatography-mass spectrometry (LC-MSMS). LPS treatment caused significant translocation of AIF and endonuclease-G in neuronal N2A cells which were significantly attenuated with piracetam co-treatment. Significant over-expression of proinflammatory cytokines was also observed after treatment of LPS to cells which was inhibited with piracetam co-treatment demonstrating its anti-inflammatory property. LPS-treated cells exhibited significant oxidative DNA fragmentation and poly [ADP-ribose] polymerase-1 (PARP-1) up-regulation in nucleus, both of which were attenuated with piracetam treatment. Antioxidant melatonin but not z-VAD offered the inhibited LPS-induced DNA fragmentation indicating the involvement of oxidative DNA fragmentation. Further, we did not observe the altered caspase-3 level after LPS treatment initially while at a later time point, significantly augmented level of caspase-3 was observed which was not inhibited with piracetam treatment. In total, our findings indicate the interference of piracetam in mitochondrion-mediated caspase-independent pathway, as well as its anti-inflammatory and antioxidative properties.

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Graphical Abstract Graphical abstract indicating the novel interference of metabolic enhancer piracetam (P) in neuronal death mechanisms

     

IgD positive L&H cells identify a unique subset of nodular lymphocyte predominant Hodgkin lymphoma

Nodular lymphocyte predominant Hodgkin lymphoma (NLPHL) is a rare B-cell lymphoma considered to be of germinal center (GC) derivation. Studies on immunoglobulin expression have been few, and post-switch immunoglobulin (IgG) has been identified in the majority of cases examined thus far. We reviewed 180 cases of NLPHL and observed the unexpected expression of IgD in 27% of cases. IgD is usually coexpressed with IgM in naive B cells but can also be seen as IgD-only in centroblasts (CD38-positive) or memory B cells (CD27-positive). We asked whether IgD-positive NLPHL differed from cases of NLPHL negative for IgD. Clinically, the IgD-positive cases presented at a younger median age (21 vs. 44 years) and had a striking male predominance (male-to-female ratio, 23:1 vs. 1.5:1). Cervical lymph nodes were more frequently involved (56% vs. 18.2%). L&H cells were localized in a predominantly extrafollicular distribution in the majority of IgD-positive cases (69%). The IgD-positive cases did not coexpress IgM or CD27 (a marker associated with memory B cells), and nearly all (93%) were weakly positive for CD38, supporting a GC derivation. The expression of Bcl-6, BOB.1, Oct2, and SWAP-70 was similar in the two groups. However, PU.1 expression was seen in 60% of the IgD-positive cases in contrast to 86% of the IgD-negative cases. The absence of PU.1 staining correlated with more L&H cells in an extrafollicular distribution, weakening the use of this marker in the differential diagnosis with T-cell rich/histiocyte rich B-cell lymphomas. To study IgD expression in "de-novo" T-cell rich/histiocyte rich B-cell lymphomas, we analyzed 20 cases and all but one were negative. In conclusion, cases of IgD-positive NLPHL do not differ from IgD-negative cases regarding cellular derivation and most other immunophenotypic characteristics. However, IgD-positive NLPHL exhibits distinctive clinical features, and more often involves the interfollicular region in a background relatively rich in T cells. IgD positivity may represent an additional useful marker in the diagnosis of NLPHL.     

Deep-seated plexiform schwannoma: a pathologic study of 16 cases and comparative analysis with the superficial variety

Plexiform schwannoma (PS) is one of the least common histologic variants of schwannoma. It shows a plexiform growth pattern and typically occurs in the dermis and subcutaneous tissue. Morphologically, PS can display a conventional, cellular, or mixed appearance. However, the frequent cellular morphology associated with hyperchromatic nuclei, increased mitoses, and plexiform growth can suggest a malignant process, mainly a high-grade malignant peripheral nerve sheath tumor (MPNST). The purpose of this study was to analyze the clinicopathologic features of deep-seated PS and compare them with the superficial counterparts. Sixteen deep-seated PSs were analyzed clinicopathologically, immunohistochemically, and ultrastructurally, and compared with 8 superficial (5 dermal and 3 subcutaneous) PSs. There were 12 females and 4 males, ranging from 5 months to 61 years of age. Fifteen tumors were located in the deep somatic soft tissue (extremities, 8; retroperitoneum/pelvis, 3; trunk, 2; parotid, 1; vulva, 1) and 1 tumor was located in the thoracic esophagus. None of the patients had stigmata of neurofibromatosis. Local recurrence was noted in half of the patients with clinical information available, but none had evidence of disease at last follow-up. Worrisome morphologic features included: increased cellularity (68%), mild to moderate pleomorphism (50%), and mitotic activity (93%) ranging from 1 to 10 MF/10 high power fields (HPFs). Focal necrosis was seen in 12% and myxoid change was identified in 18% of cases. Immunohistochemical stains for S-100 protein showed strong and diffuse positivity on all cases tested. Ultrastructurally, findings characteristic of schwannian differentiation were identified in the cases analyzed. The 8 superficial PSs showed increased cellularity and mild to moderate pleomorphism in 62% of cases but lacked tumor necrosis. Deep-seated PS is a rare, under-recognized PNST of deep soft tissue, typically not associated with neurofibromatosis. Although commonly occurring in the extremities, they can be seen in other locations including the viscera. In contrast with the more common superficial (dermal and subcutaneous) tumors, deep PSs have a predilection for females, can occur in congenital settings, and can show necrosis and myxoid change. Common worrisome histologic features seen in both groups include increased cellularity and mitoses. It is important to differentiate these tumors from plexiform neurofibromas and MPNSTs as they follow a benign clinical course, with complete surgical excision being curative.     

Comparative immunohistochemical profile of hepatocellular carcinoma,cholangiocarcinoma, and metastatic adenocarcinoma

Distinguishing hepatocellular carcinoma (HCC) from cholangiocarcinoma (CC) and metastatic adenocarcinoma (MA) involving the liver can be problematic, often requiring the use of immunohistochemistry to facilitate diagnosis. Hep Par 1, a monoclonal antibody with expression confined primarily to benign and malignant hepatocytes, has recently become commercially available. We evaluated Hep Par 1 along with other immunohistochemical markers used to differentiate HCC, CC, and MA, including AE1/AE3, CAM 5.2, B72.3, monoclonal carcinoembryonic antigen (mCEA), polyclonal CEA (pCEA), alpha-fetoprotein (AFP), factor XIIIa, inhibin, CD10, villin, MOC-31, cytokeratin (CK) 7, CK 19, and CK 20, to determine the markers most useful in differentiating these entities. Forty-two cases of HCC, 9 cases of CC, and 56 cases of MA (24 colon, 15 pancreas, 8 ovary, 5 breast, and 4 stomach) were studied. Hep Par 1 was sensitive and specific for HCC, with 38 of 42 (90%) cases staining positively, whereas reactivity was observed in only 8 of 56 (14%) MAs and 0 of 9 CCs. Though limited somewhat by poor sensitivity, a bile canalicular pattern of staining with pCEA, CD10, and villin was specific for HCC and was not observed in the other tumors. Lack of mCEA and MOC-31 immunoreactivity was also characteristic of HCCs. CK 19 positivity favored CC over HCC, but was not useful in differentiating CC from MA. Expression of AFP, although observed in only about one third of the cases, favored HCC over CC and MA. CK 7 and CK 20 were also useful in this differential diagnosis, particularly when dealing with MA of colonic origin. AE1/AE3, CAM 5.2, B72.3, inhibin, and factor XIIIa were noncontributory in differentiating these entities.