Morphometric study of the suprascapular notch and scapular dimensions in adult Malawian cadavers and implications of completely ossified superior transverse scapular ligament.

Introduction: the anatomy of the suprascapular notch and its relationship to scapular dimensions are critical in the management of suprascapular neuropathies. Individuals show considerable differences in the dimensions of the suprascapular notch across populations. The purpose of this study was to determine the morphology and morphometric dimensions of the suprascapular notch in adult Malawian cadavers and to suggest clinical implications associated with complete ossification of the suprascapular ligament. Methods: adult dry scapulae from undetermined sex specimens (n=125) obtained from the skeletal collection at Kamuzu University of Health Sciences were classified according to the Rengachary categorization method to assess the suprascapular notch superior transverse distance, mid transverse distance, depth, scapula length and width using a standard Vernier caliper. Results: the most prevalent suprascapular notch class was type I, which was found in 46 (36.8%) of all scapulae. Type VI was the least common, found in only 1 (0.8%) of the scapulae. The mean notch superior transverse distance was 1.3 ± 0.6 cm, while the mean maximum depth was 0.6 ± 0.3 cm. Only the differences in depth, however, were statistically significant (p=0.001). Conclusion: the current study has described the morphology and morphometry of the suprascapular notch in relation to the risk of suprascapular nerve entrapment associated with complete ossification of the suprascapular ligament. Our sample population generally showed smaller suprascapular notch and scapular dimensions than other populations. This should be considered during the management of suprascapular neuropathy and preoperative planning of surgical operations of the shoulder region.

In vitro effects of aqueous extract of unfermented rooibos on human spermatozoa.

The inability to conceive is a baleful experience for thousands of couples worldwide. Among different well-known reproductive techniques, medicinal plants have been utilized to treat male infertility. Medicinal plants, provide a therapeutic alternative, which is available and affordable for infertile couples. We investigated the direct effect of unfermented rooibos aqueous extract on human spermatozoa. Semen samples (n = 50) collected from donors and patients consulting for fertility were reassigned as normal (n = 22) and abnormal (n = 28) samples based on the outcome of the baseline semen analysis, using the World Health Organization (WHO) cut off value. Semen samples were allowed to liquefy and subsequently washed with human tubular fluid in bovine serum albumin medium. The samples were then treated with aqueous extracts of unfermented rooibos (0, 0.15, 1.5, 15, 150 µg/ml) at 37°C for 1 h and assessed thereafter. Sperm motility, vitality, DNA fragmentation, intracellular reactive oxygen species and mitochondrial membrane potential in both groups remained unchanged (p > 0.05). However, aqueous extract of unfermented rooibos (only at 1.5 µg/ml) significantly increased capacitation and acrosome reaction in the abnormal sample group (p > 0.05). Unfermented rooibos aqueous extract had no deleterious impact on human spermatozoa's function and might be attributed to its antioxidant properties.

In vitro effects of aqueous extract of fermented rooibos (Aspalathus linearis) on human sperm function.

Aspalathus linearis (rooibos) is a herbal medicinal plant originally from South Africa's fynbos and well known for its medicinal effects in treating different medical conditions. Rooibos contains significant levels of antioxidants capable of inhibiting the production of reactive oxygen species, which may improve seminal parameters. This study focussed on investigating the direct effect of fermented rooibos on human sperm functions in vitro. Semen samples collected by masturbation from unproven fertile donors (n = 25) and infertile patients (n = 25) after 3-5 days' abstinence were liquefied and centrifuged (300 × g; 10 min) in human tubular fluid medium containing 1% bovine serum albumin. Afterwards, semen samples (7.5 × 106 /ml) were incubated at 37°C for one hour with aqueous extract of fermented extract in sperm preparation medium (0, 0.10, 1.0, 10 and 100 µg/ml) and assessed. Our data showed that fermented rooibos did not affect functional sperm parameters (motility, vitality, intracellular reactive oxygen species and acrosome reaction, p > .05), in vitro except in the reduced percentage of intact mitochondrial membrane potential and DNA fragmentation (p < .05). The decrease in DNA fragmentation generates the possibility of using the extract in patients prior to assisted reproductive techniques.

A morphometric study of the mandibular foramen in dry adult human mandibles in a black Malawian population / Un estudio morfométrico del foramen mandibular en mandíbulas humanas adultas secas en una población malauí negra

SUMMARY: The inferior alveolar nerve block (IANB) technique is a common technique performed on patients in dental surgery, placement of mandibular implants and other procedures involving the mandible. Precise identification of the mandibular foramen (MF) is essential for dental surgeons to accurately administer local anesthetics. Inaccurate localization of the mandibular foramen may result in IANB failure and injury to neurovascular tissues. Therefore, this study aimed at investigating the precise location of the MF from various anatomical land marks in dry adult human mandibles of Malawian population. The study was conducted on 29 dry adult human mandibles of unknown sex of Malawian origin from the Anatomy Division collection of human skeletons housed in the Biomedical Sciences Department, College of Medicine, University of Malawi. To determine the position of the mandibular foramen, distances from mandibular foramen to anterior margin, posterior margin, mandibular notch, gonial angle and mandibular base using a Vernier caliper were measured. The mean distance of the MF from posterior margin of mandibular ramus was 11.26±1.22 mm (right side) and 11.47±1.35 mm (left side), from the anterior margin 20.85±3.12 mm (right side) and 20.85±3.22 mm (left side) mandibles. The mean distance between mandibular notch (MN) and MF was 23.87±2.61 mm (right side) and 23.53 ± 2.65 mm (left side). The mean distance between mandibular base (MB) and MF for the right and left were 28.47 ± 2.90 mm and 27.85 ± 2.99 mm respectively. The inferior limit of the mandibular foramen was located at 24.69 ± 3.65 mm (right side) and 24.25 ± 2.77 mm (left side) to the angle (AG) of the mandible. The findings of this study show that the anterior margin mean distance from the MF for both right and left mandibles seem to be bilateral symmetrical suggesting the interpretation that the needle for IANB could be inserted at about 21 mm from the anterior margin to the MF in an adult of Malawian origin during surgery.

RESUMEN: La técnica de bloqueo del nervio alveolar inferior (IANB) es una técnica común que se realiza en pacientes en cirugía dental, colocación de implantes mandibulares y otros procedimientos que involucran la mandíbula. La identificación precisa del foramen mandibular (MF) es esencial para que los cirujanos dentistas administren con precisión anestésicos locales. La localización inexacta del foramen mandibular puede resultar en una falla de la IANB y lesión de los tejidos neurovasculares. Por lo tanto, este estudio tuvo como objetivo investigar la ubicación precisa de la MF de varias marcas anatómicas en las mandíbulas humanas adultas secas de la población de Malawi. El estudio se llevó a cabo en 29 mandíbulas humanas adultas secas de sexo desconocido de origen malauí de la colección de esqueletos humanos de la División de Anatomía del Departamento de Ciencias Biomédicas de la Facultad de Medicina de la Universidad de Malawi. Para determinar la posición del foramen mandibular, se midieron las distancias desde el foramen mandibular hasta el margen anterior, margen posterior, incisura mandibular, ángulo gonial y base mandibular utilizando un calibre Vernier. La distancia media del MF desde el margen posterior de la rama mandibular fue de 11,26 ± 1,22 mm (lado derecho) y 11,47 ± 1,35 mm (lado izquierdo), desde el margen anterior 20,85 ± 3,12 mm (lado derecho) y 20,85 ± 3,22 mm (lado izquierdo) lado) mandíbulas. La distancia media entre la muesca mandibular (MN) y MF fue de 23,87 ± 2,61 mm (lado derecho) y 23,53 ± 2,65 mm (lado izquierdo). La distancia media entre la base mandibular (MB) y MF para la derecha y la izquierda fue de 28,47 ± 2,90 mm y 27,85 ± 2,99 mm, respectiva- mente. El límite inferior del foramen mandibular se ubicó a 24,69 ± 3,65 mm (lado derecho) y 24,25 ± 2,77 mm (lado izquierdo) del ángulo (AG) de la mandíbula. Los resultados de este estudio mues- tran que la distancia media del margen anterior desde el MF para las mandíbulas derecha e izquierda parece ser simétrica bilateral, lo que sugiere la interpretación de que la aguja para IANB podría insertarse a unos 21 mm del margen anterior al MF en una adulto de origen malauí durante la cirugía.

Lopinavir and curcumin directly alters BAX/BCL2 and VEFG165b mRNA levels to suppress human squamous cervical carcinoma cell growth / Lopinavir y curcumina alteran directamente los niveles de ARNm de BAX/ BCL 2 y VEFG165 para suprimir el crecimiento de células de carcinoma cervical escamoso humano

Following the success of the highly active antiretroviral therapy, the potential of multidrug combination regimen for the management of cancer is intensely researched. The anticancer effects of curcumin on some human cell lines have been documented. Lopinavir is a FDA approved protease inhibitor with known apoptotic activities. Dysregulated apoptosis is important for the initiation of cancer while angiogenesis is required for cancer growth and development, this study therefore investigated the effects of the combination of lopinavir and curcumin on cell viability, apoptosis and the mRNA expression levels of key apoptotic and angiogenic genes; BAX, BCL2 and VEGF165b in two human cervical cell lines; human squamous cell carcinoma cells - uterine cervix (HCS-2) and transformed normal human cervical cells (NCE16IIA). The two human cervical cell lines were treated with physiologically relevant concentrations of the agents for 120 h following which BAX, BCL2 and VEGF165b mRNA expression were determined by Real Time qPCR. The Acridine Orange staining for the morphological evaluation of apoptotic cells was also performed. The combination of lopinavir and curcumin up-regulated pro-apoptotic BAX and antiangiogenic VEGF165b but down-regulated the mRNA levels of anti-apoptotic BCL2 mRNA in the human squamous cell carcinoma (HCS-2) cells only. The fold changes were statistically significant. Micrographs from Acridine Orange staining showed characteristic evidence of apoptosis in the human squamous cell carcinoma (HCS-2) cells only. The findings reported here suggest that the combination of curcumin and the FDA approved drug-lopinavir modulate the apoptotic and angiogenic pathway towards the inhibition of cervical cancer.

Tras el éxito de la terapia antirretroviral altamente activa, se investiga intensamente el potencial del régimen de combinación de múltiples fármacos para el tratamiento del cáncer. Se han documentado los efectos anticancerígenos de la curcumina en algunas líneas celulares humanas. Lopinavir es un inhibidor de proteasa aprobado por la FDA con actividades apoptóticas conocidas. La apoptosis disrregulada es importante para el inicio del cáncer, mientras que la angiogénesis es necesaria para el crecimiento y desarrollo del cáncer. Por lo tanto, este estudio investigó los efectos de la combinación de lopinavir y curcumina sobre la viabilidad celular, la apoptosis y los niveles de expresión del ARNm de genes apoptóticos y angiogénicos clave: BAX, BCL2 y VEGF165b en dos líneas celulares cervicales humanas; células de carcinoma de células escamosas humanas: cérvix uterino (HCS-2) y células cervicales humanas transformadas (NCE16IIA). Las dos líneas celulares cervicales humanas se trataron con concentraciones fisiológicamente relevantes de los agentes durante 120 horas, después de lo cual la expresión de ARNm de BAX, BCL2 y VEGF165b se determinó mediante qPCR en tiempo real. También se realizó la tinción con naranja de acridina para la evaluación morfológica de células apoptóticas. La combinación de lopinavir y curcumina reguló incrementando BAX proapoptósicos y VEGF165b antiangiogénicos, pero reguló a la baja los niveles de ARNm del BCL2 antiapoptótico en células de carcinoma de células escamosas humanas (HCS-2) únicamente. Los cambios en el pliegue fueron estadísticamente significativos. Las micrografías de la tinción con naranja de acridina mostraron evidencia característica de apoptosis solo en las células del carcinoma de células escamosas humanas (HCS-2). Los hallazgos reportados aquí sugieren que la combinación de curcumina y el fármaco aprobado por la FDA lopinavir modulan la vía apoptótica y angiogénica hacia la inhibición del cáncer cervical.

Antiangiogenic VEGF165b expression in human breast MCF-7 and MCF-10A cells exposed to reverse transcriptase and protease inhibitors / Expresión de VEGF165b antiangiogénico en células MCF-7 y MCF-10A de mama humana expuesto a inhibidores de la transcriptasa inversa y la proteasa

The combined antiretroviral therapy (cART), a multidrug combination regimen, usually consisting Nucleoside Reverse Transcriptase Inhibitors, non- Nucleoside Reverse Transcriptase Inhibitors and Protease Inhibitors has altered the morbidity pattern affecting HIV-infected individuals to include non-AIDS-defining malignancies (nADMs). The speculation is rife; does cART induce or promote the progression of nADMs such as breast cancer? This study was therefore designed to investigate of the effects of some antiretroviral drugs (at clinically relevant concentrations) on the expression of anti-angiogenic gene; VEGF165b in two human breast cell lines; MCF-7 and MCF-10A by Real Time qPCR and immuno-fluorescence. All of the antiretroviral drugs and combinations tested produced patterns of slight up or downregulation of VEGF165b mRNA expression but the alterations did not attain statistical significance. They also did not alter VEGF165bprotein localisation in both cell lines. The findings reported here suggest that antiretroviral drugs probably do not influence the angiogenic pathway in the development of breast cancer in patients under the combined antiretroviral regimen.

El tratamiento antirretroviral combinado (TARc), un régimen de combinación de múltiples fármacos, consistiendo generalmente en inhibidores nucleósidos de la transcriptasa reversa, inhibidores no-nucleósidos de la transcriptasa reversa e inhibodres de proteasa que alteran el patrón de mortalidad que afecta a infectados por el VIH incluyendo neoplasias definidas como no HIV (nADMs). La especulación es moneda corriente; TARc induce o promueve la progresión de nADMs como cáncer de mama? Por lo tanto, este estudio se diseñó para investigar los efectos de algunos de los fármacos antirretrovirales (en concentraciones clínicamente relevantes) sobre la expresión del gen anti-angiogénico; VEGF165b en dos líneas celulares de mama humana; MCF-7 y MCF-10A por PCR tiempo real e inmunofluorescencia. Todos los fármacos antirretrovirales y las combinaciones probadas pueden regular en forma ligera hacia arriba o hacia abajo la expresión de ARNm producidos por VEGF165b pero las alteraciones no fueron estadísticamente significativos. Además, no se alteran los niveles de proteína VEGF165b, para la localización en ambas líneas celulares. Los resultados aquí presentados sugieren que los medicamentos antirretrovirales probablemente no influyen en la vía angiogénica en el desarrollo del cáncer de mama en pacientes bajo el régimen antirretroviral combinado.

BAX/BCL-2 mRNA and protein expression in human breast MCF-7 cells exposed to drug vehicles-methanol and dimethyl sulfoxide (DMSO) for 24 hrs.

BACKGROUND: Methanol and DMSO are commonly used as carrier solvents for lipophilic chemicals in in-vitro experiments. However, very little information is available regarding the effects of these solvents on the expression of pro and anti-apoptotic genes and proteins. MATERIALS AND METHODS: In this study, we examined the cytotoxic effects of methanol and dimethylsulfoxide at 0.5% (final concentrations recommended for in-vitro toxicity assays) on human breast cancer MCF-7 cells. We also investigated the effects of these solvents on the mRNA and immunocytochemical expression of apoptotic proteins BAX and BCL-2. RESULTS: The results of neutral red cell viability assay showed that methanol and DMSO concentrations of 0.5% exhibited no cytotoxic effects on MCF-7 cells following a 24 hour exposure. Gene expression and Immunofluorescence results showed that methanol but not DMSO reduced the expression of the BAX pro-apoptotic protein, while both solvents did not alter the expression of the BCL-2 oncoprotein. CONCLUSION: Our results suggest that while methanol concentrations at 0.5% may be appropriate for in vitro toxicity studies in human breast cancer MCF-7 cells, it could alter the results of gene and protein expression experiments.

The effects of HAART on the expression of MUC1 and P65 in a cervical cancer cell line, HCS-2.

Cervical cancer is the third most commonly diagnosed cancer globally and it is one of three AIDS defining malignancies. Highly active antiretroviral therapy (HAART) is a combination of three or more antiretroviral drugs and has been shown to play a significant role in reducing the incidence of some AIDS defining malignancies, although its effect on cervical cancer is still unclear. The aim of this study was to investigate the relationship between cervical cancer and HAART. This was achieved by studying the expression of two signalling molecules expressed in cervical cancer; MUC1 and P65. Following the 24-hour treatment of a cervical cancer cell line, HCS-2, with drugs, which are commonly used as part of HAART at their clinical plasma concentrations, real-time qPCR and immunofluorescence were used in order to study gene and protein expression. A one-way ANOVA followed by a Tukey-Kramer post-hoc test was conducted using JMP 11 software on both sets of data. The drug classified as a protease inhibitor (PI) (i.e. LPV/r) reduced MUC1 and P65 gene and protein expression more than the other drug tested. PIs are known to play a significant role in cell death; therefore, the cells were thought to be more susceptible to cell death following treatment with PIs. In conclusion, the drugs used, especially the PI showed some anticancer effects by facilitating cell death through decreased gene and protein expression of MUC1 and P65 and present promising agents for cancer treatment.

Effects of HIV protease, nucleoside/non-nucleoside reverse transcriptase inhibitors on Bax, Bcl-2 and apoptosis in two cervical cell lines.

Protease inhibitors (PIs) and reverse transcriptase drugs are important components of highly active antiretroviral therapy (HAART) for treating human acquired immunodeficiency syndrome (AIDS). Long-term clinical therapeutic efficacy and treatment compliance of these agents have been limited by undesirable adverse effects and their oncogenicity has been queried. This study investigated the effects of selected antiretroviral agents on the expression of key apoptotic regulatory genes; Bax and Bcl-2 in two cervical cell lines HCS-2 and NCE16IIA by real-time qPCR gene expression and immunocytochemistry. The anti-apoptotic effects of the PI-LPV/r were investigated by cell death detection ELISA and acridine orange staining. All the antiretroviral drugs and combinations tested had no effects on Bax and Bcl-2 gene expression and protein localisation in both cell lines. The protease inhibitors-LPV/r exhibited significant (P<0.05) inhibition of camptothecin-induced apoptosis in the cervical cancer HCS-2 cell line but not in the normal immortalised NCE16IIA cell line. This anti-apoptotic property of HIV protease inhibitors, although shown so far not to involve protein and RNA synthesis might promote the development of cancer.