Correction: Jose et al. Transferrin-Conjugated Docetaxel-PLGA Nanoparticles for Tumor Targeting: Influence on MCF-7 Cell Cycle. Polymers 2019, 11, 1905.

In the original publication [...].

The Immunomodulatory Effects of Honey and Associated Flavonoids in Cancer.

Honey has exerted a high impact in the field of alternative medicine over many centuries. In addition to its wound healing, anti-microbial and antioxidant properties, several lines of evidence have highlighted the efficiency of honey and associated bioactive constituents as anti-tumor agents against a range of cancer types. Mechanistically, honey was shown to inhibit cancer cell growth through its pro-apoptotic, anti-proliferative and anti-metastatic effects. However, the potential of honey to regulate anti-tumor immune responses is relatively unexplored. A small number of in vitro and in vivo studies have demonstrated the ability of honey to modulate the immune system by inducing immunostimulatory as well as anti-inflammatory effects. In the present review, we summarize the findings from different studies that aimed to investigate the immunomodulatory properties of honey and its flavonoid components in relation to cancer. While these studies provide promising data, additional research is needed to further elucidate the immunomodulatory properties of honey, and to enable its utilization as an adjuvant therapy in cancer.

Ambrisentan, an endothelin receptor type A-selective antagonist, inhibits cancer cell migration, invasion, and metastasis.

Several studies reported a central role of the endothelin type A receptor (ETAR) in tumor progression leading to the formation of metastasis. Here, we investigated the in vitro and in vivo anti-tumor effects of the FDA-approved ETAR antagonist, Ambrisentan, which is currently used to treat patients with pulmonary arterial hypertension. In vitro, Ambrisentan inhibited both spontaneous and induced migration/invasion capacity of different tumor cells (COLO-357 metastatic pancreatic adenocarcinoma, OvCar3 ovarian carcinoma, MDA-MB-231 breast adenocarcinoma, and HL-60 promyelocytic leukemia). Whole transcriptome analysis using RNAseq indicated Ambrisentan's inhibitory effects on the whole transcriptome of resting and PAR2-activated COLO-357 cells, which tended to normalize to an unstimulated profile. Finally, in a pre-clinical murine model of metastatic breast cancer, treatment with Ambrisentan was effective in decreasing metastasis into the lungs and liver. Importantly, this was associated with a significant enhancement in animal survival. Taken together, our work suggests a new therapeutic application for Ambrisentan in the treatment of cancer metastasis.

Therapeutic and preventive properties of honey and its bioactive compounds in cancer: an evidence-based review.

Despite the much improved therapeutic approaches for cancer treatment that have been developed over the past 50 years, cancer remains a major cause of mortality globally. Considerable epidemiological and experimental evidence has demonstrated an association between ingestion of food and nutrients with either an increased risk for cancer or its prevention. There is rising interest in exploring agents derived from natural products for chemoprevention or for therapeutic purposes. Honey is rich in nutritional and non-nutritional bioactive compounds, as well as in natural antioxidants, and its potential beneficial function in human health is becoming more evident. A large number of studies have addressed the anti-cancer effects of different types of honey and their phenolic compounds using in vitro and in vivo cancer models. The reported findings affirm that honey is an agent able to modulate oxidative stress and has anti-proliferative, pro-apoptotic, anti-inflammatory, immune-modulatory and anti-metastatic properties. However, despite its reported anti-cancer activities, very few clinical studies have been undertaken. In the present review, we summarise the findings from different experimental approaches, including in vitro cell cultures, preclinical animal models and clinical studies, and provide an overview of the bioactive profile and bioavailability of the most commonly studied honey types, with special emphasis on the chemopreventive and therapeutic properties of honey and its major phenolic compounds in cancer. The implications of these findings as well as the future prospects of utilising honey to fight cancer will be discussed.

Transferrin-Conjugated Docetaxel-PLGA Nanoparticles for Tumor Targeting: Influence on MCF-7 Cell Cycle.

Targeted drug delivery systems are commonly used to improve the therapeutic index of anti-cancer drugs by increasing their selectivity and reducing systemic distribution and toxicity. Ligand-conjugated nanoparticles (NPs) can be effectively applied for active chemotherapeutic targeting to overexpressed receptors of tumor cells. In this study, transferrin (Tf) was successfully conjugated with poly-l-lactic-co-glycolic acid (PLGA) using ethylene diamine confirmed by NMR, for the loading of docetaxel trihydrate (DCT) into PLGA nanoparticles (NPs). The DCT-loaded Tf-conjugated PLGA NPs were produced by an emulsion-solvent evaporation technique, and a 32 full factorial design was used to optimize the nanoparticle formulations. The DCT-loaded Tf-conjugated PLGA NPs were characterized by FTIR spectroscopy, differential scanning calorimetry, powder X-ray diffraction (PXRD), TEM, particle size, and zeta potential analysis. In vitro release kinetics confirmed that release of DCT from the designed formulations followed a zero-order kinetics and a diffusion controlled non-Fickian release profile. The DCT-loaded Tf-conjugated PLGA NPs were evaluated in vitro in MCF-7 cells for bioactivity assessment. Cytotoxicity studies confirmed that the Tf-conjugated PLGA NPs were more active than the non-conjugated counterparts. Cell uptake studies re-confirmed the ligand-mediated active targeting of the formulated NPs. From the cell cycle analysis, the anti-cancer activity of DCT-loaded Tf-conjugated PLGA NPs was shown to occur by arresting the G2/M phase.

Inhibition of Tyrosine-Phosphorylated STAT3 in Human Breast and Lung Cancer Cells by Manuka Honey is Mediated by Selective Antagonism of the IL-6 Receptor.

Aberrantly high levels of tyrosine-phosphorylated signal transducer and activator of transcription 3 (p-STAT3) are found constitutively in ~50% of human lung and breast cancers, acting as an oncogenic transcription factor. We previously demonstrated that Manuka honey (MH) inhibits p-STAT3 in breast cancer cells, but the exact mechanism remained unknown. Herein, we show that MH-mediated inhibition of p-STAT3 in breast (MDA-MB-231) and lung (A549) cancer cell lines is accompanied by decreased levels of gp130 and p-JAK2, two upstream components of the IL-6 receptor (IL-6R) signaling pathway. Using an ELISA-based assay, we demonstrate that MH binds directly to IL-6Rα, significantly inhibiting (~60%) its binding to the IL-6 ligand. Importantly, no evidence of MH binding to two other cytokine receptors, IL-11Rα and IL-8R, was found. Moreover, MH did not alter the levels of tyrosine-phosphorylated or total Src family kinases, which are also constitutively activated in cancer cells, suggesting that signaling via other growth factor receptors is unaffected by MH. Binding of five major MH flavonoids (luteolin, quercetin, galangin, pinocembrin, and chrysin) was also tested, and all but pinocembrin could demonstrably bind IL-6Rα, partially (30-35%) blocking IL-6 binding at the highest concentration (50 µM) used. In agreement, each flavonoid inhibited p-STAT3 in a dose-dependent manner, with estimated IC50 values in the 3.5-70 µM range. Finally, docking analysis confirmed the capacity of each flavonoid to bind in an energetically favorable configuration to IL-6Rα at a site predicted to interfere with ligand binding. Taken together, our findings identify IL-6Rα as a direct target of MH and its flavonoids, highlighting IL-6R blockade as a mechanism for the anti-tumor activity of MH, as well as a viable therapeutic target in IL-6-dependent cancers.

An appraisal of cinnamyl sulfonamide hydroxamate derivatives (HDAC inhibitors) for anti-cancer, anti-angiogenic and anti-metastatic activities in human cancer cells.

Multiple genetic mutations along with unusual epigenetic modifications play a major role in cancer development. Histone deacetylase (HDAC) enzyme overexpression observed in the majority of cancers is responsible for tumor suppressor gene silencing and activation of proto-oncogenes to oncogenes. Cinnamic acid derivatives exhibit anti-cancer potential through HDAC enzyme inhibition. We have synthesized a few cinnamyl sulfonamide hydroxamate derivatives (NMJ-1, -2 and -3) by already published in-house procedures and their purity, and chemical characterization were performed by NMR, mass spectrometry and elemental analysis. The anti-cancer activities were also evaluated against colon cancer. The rationale for synthesis was based on bioisosterism concept. To take the work forward, these compounds were considered for in vitro anti-angiogenic and anti-metastatic activities in cancer cells. The effectiveness of these compounds was determined by SRB assay. The compounds showed cancer cell cytotoxicity (IC50 range of 5.7 ± 0.43 to 20.5 ± 1.9 µM). The mechanism of compound-induced cell death involves an intrinsic apoptosis pathway which was supported by the following: increase in apoptotic index, arrest in cell cycle at G2/M phase, increase in annexin V binding and induction of p21(Waf1/Cip1) expression in the treated cells. Further, their target modulating effect, measured as the expression of acetyl-H3 histone and acetyl α-tubulin was determined by Western blots. Hyper acetylation of H3 histone and α-tubulin were observed. Furthermore, increased expression of cleaved caspase-3, cleaved PARP, total Bad was estimated by ELISA. The anti-angiogenic effect was examined through cobalt (II) chloride (CoCl2)-induced HIF-1α expression, where the compounds reduced the expression of induced HIF-1α. In addition, their anti-metastatic ability was determined through phorbol-12-myristate-13-acetate (PMA)-induced expression of MMP-2 and -9 by Western blotting and gelatin zymography. Inhibition of malignant cell migration was assessed by scratch wound assay. The compounds showed a decrease in cell migration and inhibition of induced MMP-2 and MMP-9 expression. NMJ-2 exhibited comparable activity to that of standard SAHA. Our findings indicate that NMJ series of compound have potent in vitro anti-cancer, anti-angiogenic and anti-metastatic activity through HDAC enzyme inhibition.

In vitro mechanistic and in vivo anti-tumor studies of Glycosmis pentaphylla (Retz.) DC against breast cancer.

ETHNOPHARMACOLOGICAL RELEVANCE: Glycosmis pentaphylla (Retz.) DC (Rutaceae) has been traditionally used for the treatment of rheumatism, cancer, liver disorders, inflammation etc. AIM OF THE STUDY: The present study is aimed at elucidating the effect of Glycosmis pentaphylla (Retz.) DC on the key markers of apoptosis, metastasis and angiogenesis, in vitro. The study also evaluated the effect of fractions in vivo in DMBA-induced mammary tumor model. MATERIALS AND METHODS: Fractions of Glycosmis pentaphylla (Retz.) DC leaf extracts was studied for their effect on apoptotic markers in breast cancer cell lines, MCF-7 and MDA-MB-231 cells. They were also studied for their effect on metastatic and angiogenic markers, MMP-9 and HIF-1α in MCF-7 cells. The fractions were studied in vivo in DMBA-induced mammary tumor model in Sprague Dawley rats. RESULTS: The studies showed that the fractions induced apoptosis in breast cancer cells through the intrinsic/mitochondrial apoptotic pathway. The fractions were also able to inhibit the metastatic and angiogenic markers, MMP-9 and HIF-1α. Anti-tumor studies in DMBA-induced mammary model in Sprague Dawley rats also showed favorable results.

Glycosmis pentaphylla (Retz.) DC arrests cell cycle and induces apoptosis via caspase-3/7 activation in breast cancer cells.

ETHNOPHARMACOLOGICAL RELEVANCE: Glycosmis pentaphylla (Retz.) DC belonging to the family Rutaceae has been traditionally used for the treatment of rheumatism, anaemia, jaundice, skin diseases, bronchitis etc. The plant is traditionally considered as anti-cancer medicine and used by the healers of Bangladesh to treat all types of cancers. Perhaps the key to many of its medicinal applications is its inherent anti-inflammatory property. AIM OF THE STUDY: The present study is aimed at evaluating the effect of various fractions of G. pentaphylla (Retz.) DC leaves on the cell cycle and apoptosis of breast cancer cells viz. MCF-7 and MDA-MB-231. MATERIALS AND METHODS: Various extracts and fractions of the leaves of G. pentaphylla (Retz.) DC were studied for their cytotoxicity with the help of Sulforhodamine B assay, in MCF-7, MDA-MB-231 and Vero cell lines. The most active fractions were studied for their effect on the cell cycle of MCF-7 and MDA-MB-231 cells. Apoptotic studies were done using Hoechst staining, DNA fragmentation, Annexin V staining and caspase-3/7 activation assay in breast cancer cells. HPLC and HPTLC profiling of the active fractions were done. RESULTS: HPTLC and HPLC profiling revealed the presence of lupeol, chrysin, quercetin, ß-sitosterol and kaempferol as components in active fractions. Lupeol and chrysin are being reported in this plant for the first time. The studies showed that the selected fractions possess cell cycle inhibitory and apoptosis inducing effect on both MCF-7 and MDA-MB-231 cells. Apoptotic effect of the fractions on MCF-7 and MDA-MB-231 cells may be through the mitochondrial pathway by the activation of caspase-3/7.

In vitro and in vivo evaluation of novel cinnamyl sulfonamide hydroxamate derivative against colon adenocarcinoma.

The potential of cinnamic acid as an anti-inflammatory and anti-cancer agent has been studied previously. In our investigation, novel bio-isosters of cinnamyl sulfonamide hydroxamate were synthesized, characterized and confirmed for their structure and evaluated for cytotoxicity. Three NCEs namely, NMJ-1, -2 and -3 showed cell-growth inhibition in 6 human cancer cell lines with IC50 at the range of 3.3±0.15-44.9±2.6 µM. The hydroxamate derivatives of cinnamyl sulfonamide are reported inhibitors of HDAC enzyme. Thus, the effectiveness of these molecules was determined by whole cell HDAC assay in HCT 116 cell line. NMJ-2 (0.41±0.01 µM) exhibited better enzyme inhibition (IC50) compared to SAHA (2.63±0.07). In order to evaluate induction of apoptosis by treatment, Hoechst 33342 and AO/EB nuclear staining methods were used. Further, cell cycle analysis, Annexin V binding and caspase 3/7 activation assays were performed by flow cytometry where NMJ-2 significantly arrested the cell cycle at G2/M phase, increased Annexin V binding to the cell surface and activation of caspase-3/7. Bax/Bcl-2 ratio was observed by Western blot and showed an increase with NMJ-2 treatment. This was comparable to standard SAHA. The acute toxicity study (OECD-425) showed that NMJ-2 was safe up to 2000 mg/kg in rats. 1,2-Dimethyl hydrazine (DMH) was used to produce experimental colon adenocarcinoma in Wistar rats. 5-FU and NMJ-2 (100 mg/kg p.o. and 10 mg/kg i.p. once daily for 21 days, respectively) were administered to the respective groups. Both treatments significantly reduced ACFs, adenocarcinoma count, TNF-α, IL-6, nitrite and nitrate levels in colonic tissue. Our findings indicate that NMJ-2 has potent anti-cancer activity against colon cancer, by acting through HDAC enzyme inhibition and activation of intrinsic mitochondrial apoptotic pathway, with additional anti-inflammatory activity.

Intradural extramedullary primary hydatid cyst of the spine: a case report and review of literature.

Primary intradural extramedullary hydatid cyst is a rare form of parasitic infection, causing focal neurological signs, commonly observed in sheep-raising areas of the world. We report a rare case of intradural, extramedullary spinal cyst, which we had misdiagnosis in the first surgery, because of rarity of the case. A 55-year-old man presented to our hospital in August 2008. He was admitted to our clinic because of lumbar pain of increasing severity and progressive difficulty with walking and stiffness of both lower limbs, which had lasted for 1 month. On the basis of imaging results, arachnoid cyst of the lumbar spine was diagnosed. Due to rapid progression of the patient's symptoms toward spastic paraplegia, he underwent an emergency surgical decompression procedure. The patient underwent exploratory surgery using a posterior approach. A L1-L2 laminectomy was performed. After opening the dura, an intradural extramedullary cystic mass was determined. The surgical specimen measured 6 × 2 cm and was described as a whitish, pearl-like, semitranslucent, cystic material, which was thought to be parasitic. Surgery has to be followed by albendazole therapy.

A review of the tragal pointer: anatomy and its importance as a landmark in surgical procedures.

The tragal pointer has long been used as a surgical landmark for the identification of the facial nerve trunk and the maxillary artery in such procedures as parotidectomy, internal fixation of subcondylar and condylar fractures, mandibular osteotomy, temporomandibular joint arthroplasty, and percutaneous blocks of branches of the trigeminal nerve and pterygopalatine ganglion. Aside from its use as an external landmark, it has also been implicated as a contributor to crease formation in the presence of peripheral arterial disease. This article will review the available literature on the tragal pointer's use as an external landmark.

Unexplained pulmonary hypertension in peritoneal dialysis and hemodialysis patients.

OBJECTIVES: To compare the prevalence of unexplained pulmonary artery hypertension (PAH) in hemodialysis (HD) and peritoneal dialysis (PD) patients and to compare laboratory parameters between patients with unexplained PAH and those with normal pulmonary artery pressure (PAP). METHODS: We retrospectively reviewed the medical records of 278 chronic HD and 145 chronic PD patients. Laboratory findings including hemoglobin, calcium, phosphorus, alkaline phosphatase, albumin, parathyroid hormone level, serum iron, total iron binding capacity, ferritin, creatinine and blood urea nitrogen were documented. The results of transthoracic Doppler echocardiography were used to determine the pulmonary artery pressure (PAP). PAH was defined as a systolic pulmonary artery pressure (SPAP) ≥35 mmHg. To rule out secondary PAH, patients with cardiac disease, pulmonary disease, collagen vascular disease, volume overload at the time of echocardiography and positive human immunodeficiency virus test were excluded. RESULTS: Data from 34 patients in group HD and 32 individuals in group PD were analyzed. The median age of the study population was 57 (45-68) years. The median SPAP value in patients with PAH was 37.5 (35-45)mmHg. According to the echocardiographic findings, PAH was found in 14 (41.1%) patients of HD group and in 6 (18.7%) patients of PD group (P=0.04). The median serum iron and hemoglobin was significantly lower in patients with PAH compared to those in patients with normal PAP (P<0.05). CONCLUSION: Unexplained PAH seems to be more frequent in patients undergoing HD than patients in PD group. Moreover, hemoglobin and serum iron levels are lower in patients with PAH compared to those in normal PAP group.

A rare case report of subscapular artery.

Axillary artery is one of the most important arteries of the upper limb, which is a continua- tion of the subclavian artery. It begins at the lateral border of the first rib and ends at the inferior border of the teres major where it becomes the brachial artery. Axillary artery has six important branches included: 1) Superior thoracic artery 2) Thoracoacromial artery 3) Lateral thoracic artery 4) Subscapular artery 5) Posterior circumflex humeral artery 6) Anterior circumflex humeral artery. Subscapular artery arises from the third part of axillary artery normally and then divides into cir- cumflex scapular artery that extremely enters the triangular space. The other branch of subscapular artery, the thoracodorsal artery, accompanies thracodorsal nerve to lateral border of scapula and supplies and innervates that region. In this case the subscapular artery was absent in both sides and instead of that the circumflex scapular artery was directly derived from axillary artery and the thoracodorsal artery is separated from circumflex scapular artery as a thin and short branch, too. It seemed that the lateral thoracic artery, which was thicker than its normal condition, supplied the muscles of the lateral part of scapula and the thoracodorsal muscle. Other branches of the axillary artery demonstrated without any abnormally. Since axillary artery has the highest rate of rapture and damage coming after the popliteal artery, knowing the variations is important and essential for surgeons, radiologist and anatomist.

A review of the T2 segment of the brachial plexus.

Although the complex architecture of the brachial plexus (BP) has been described for decades, recent literature still aims to elucidate the variation in nerve root contributions to the BP. Understanding this variability in the nerve morphology of the BP may assist physicians and surgeons in the diagnosis and management of certain clinical conditions that involve the BP, either directly or indirectly due to its close association with neighbouring structures. In this article, we review the current anatomical knowledge of the BP, focusing especially on its T2 contribution, and discuss the causes and consequences of some relevant BP pathologies.

Refined and simplified surgical landmarks for the MacCarty keyhole and orbitozygomatic craniotomy.

BACKGROUND: Precise placement of the MacCarty keyhole, a burr hole simultaneously exposing the anterior cranial fossa floor and orbit, provides accurate, efficient entry for orbitozygomatic and supraorbital craniotomies. To locate the optimal keyhole site, previous studies have used superficial landmarks that, in our experience, are not always visible or consistent on older crania. OBJECTIVE: Therefore, we present a technique for accurate keyhole placement using landmarks that are easily visible across age ranges. METHODS: From inside the cranium, 1-mm burr holes were placed along the anterior junction of the floor and lateral wall of the anterior cranial fossa in 50 adult skulls (100 sides, with calvaria removed). Additionally, from inside the orbit, 1-mm burr holes were placed into the lateral orbital roof. Exit sites of intracranial and intraorbital burr holes were referenced to the frontozygomatic suture. The center of the site between the exiting intracranial and intraorbital holes was deemed the best location for the keyhole. RESULTS: The keyhole center was 6.8 mm (mean) superior and 4.5 mm (mean) posterior to the frontozygomatic suture, which was easily identified on all specimens. Although this keyhole center was slightly more superior on right sides than left, this was not statistically significant. In a minority of specimens, the keyhole was located near the meningo-orbital foramen (22%) and the lateral extent of the frontal sinus (2%). CONCLUSIONS: We defined an alternative method for locating the MacCarty keyhole, based on a reliable external landmark, approximately 7 mm superior and 5 mm posterior to the frontozygomatic suture.

Anatomy of the reflected ligament of the inguinal region.

BACKGROUND: There is a paucity in the literature regarding the reflected ligament. Therefore, the present study was performed in order to further elucidate this anatomy. MATERIAL AND METHODS: Eighteen formalin-fixed adult cadavers (35 sides) underwent dissection of the medial inguinal region. The reflected ligament was observed for and when identified, its dimensions were measured. RESULTS: 83% of sides were found to have a reflected ligament. These were identified in 16 male and 13 female bodies. The size and shape for the reflected ligaments were variable but overall, triangular in nature. In general, the reflected ligament was found to extend from the lacunar and medial inguinal ligaments and extended obliquely toward the midline at an approximate 45 degrees angle to insert near the linea alba. Two ligaments (6.9 %) were identified that interdigitated with the contralateral reflected ligament. The medial and lateral lengths of the ligament had a mean measurement of 2.28 and 2.58 cm. The base of the reflected ligament had a mean of 2.52 cm and the height of this ligament was found to have a mean of 2.56 cm. The mean area of the reflected ligament was calculated as 2.93 cm(2). There was no statistically significant difference between right or left sides or between genders. CONCLUSIONS: The reflected ligament was identified in the majority of our specimens and this structure usually contributed to the formation of the posteromedial wall of the external inguinal ring. Therefore, this fact should be included in future descriptions of this ligament.

An interrupted inferior vena cava in a situs inversus. Case report and review of the literature.

Situs inversus with interrupted inferior vena cava is an uncommon anatomic variant found in the abdominal and thoracic viscera. In this report, we present a 59-year-old woman with this variation, found during gross anatomical dissection. While this type of variation has been variable, in the present case the hepatic veins drained directly into a very short (2.2 cm) inferior vena cava. The infrarenal component of the inferior vena cava was present and drained into the azygos and hemiazygos veins. Clinical considerations of this variant anatomy are of interest, as they may present in patients as pathology on cross sectional imaging.

Can Doppler ultrasonographic indices of the renal artery predict the presence of supernumerary renal arteries?

BACKGROUND: Variations of the renal arteries, including the presence of supernumerary renal arteries, are important to be identified prior to renal transplant. Angiography has been the gold standard test for the pretransplant evaluation of the renal vasculature. However, this modality is expensive and invasive. The aim of this study was to assess whether Doppler ultrasonographic (DU) indices of the renal artery could predict the presence of supernumerary renal arteries. METHODS AND MATERIALS: Retrospectively, we analyzed multidetector computed tomography angiography (presence or absence of the supernumerary renal artery), DU (peak systolic velocity, resistive index, pulsatility index, end-diastolic velocity, and acceleration time) findings of 30 healthy potential renal transplant donors. Recipient operator characteristic (ROC) curves were used to examine the predictive values of the available DU indices for supernumerary renal arteries. RESULTS: The mean age of donors was 28.4 +/- 4.1 years. Of 60 kidneys evaluated, a supernumerary renal artery was found in 10%. The ROC curve analysis revealed an area under the curve of noninformative (below 0.5) for all DU parameters, indicating that none of the studied parameters could predict the presence of a supernumerary renal artery. CONCLUSIONS: Although the smaller diameter of the main renal artery has previously been found to predict the presence of supernumerary renal arteries, the present study revealed that DU indices of the renal artery may not indicate the presence of supernumerary renal arteries.