Prediction of the extent of germ cell loss utilising a noninvasive spectroscopy method in rat testicular damage model.

The aim of this study is to investigate the efficiency of elastic light single-scattering spectroscopy system, a noninvasive method, to acquire spectra during testicular biopsy from normal and damaged seminiferous tubules with various degrees of germ cell loss. Adult control rats and doxorubicin-injected rats to achieve seminiferous germ cell loss (for 10 days [10D], 20 days [D20], 30 days [D30], 40 days [D40], and 50 days [D50]) were used. Spectroscopic measurements were acquired utilising a single-fibre optical probe, and histopathology of the biopsied testicular tissue samples were compared. Time-dependent testicular damage comprising various degrees of seminiferous tubule degeneration after doxorubicin-administration was observed. In D30, D40 and D50 groups, where significant germ cell loss was identified, elastic light single-scattering spectroscopy system signals were well correlated with disturbed spermatogenesis where significant differences in spectral signals were obtained. Our findings indicate that the elastic light single-scattering spectroscopy system has the potential to enable instant imaging of spermatogenesis in rats and could also be useful in humans for clinical applications, such as to increase sperm recovery success during micro-TESE for men with nonobstructive azoospermia.

Noninvasive assessment of oral lesions using elastic light single-scattering spectroscopy: a pilot study.

PURPOSE: In the present study, we investigated the potential application of elastic light single-scattering spectroscopy (ELSSS) as a noninvasive, adjunctive tool to differentiate between malignant and benign oral lesions in vivo. METHODS: ELSSS spectra were acquired from 52 oral lesions of 47 patients prior to surgical biopsy using a single optical fiber probe. The sign of the spectral slope was used as a diagnostic parameter and was compared to the histopathology findings to obtain sensitivity and specificity of the ELSSS system in differentiating between benign and malignant tissues. RESULTS: The sign of the spectral slope was positive for the benign tissues and negative for the malignant tissues. Nine malignant lesions and one high-grade dysplasia were correctly classified as cancerous. Six out of the ten low-grade dysplasia were correctly classified as cancerous, and four of them were misclassified as benign. Thirty benign lesions were correctly classified as benign, and two were misclassified as malignant. Our results indicate that the sign of the spectral slope enables the differentiation between malignant and benign oral lesions with a sensitivity of 80% and specificity of 94%. CONCLUSIONS: ELSSS has the potential to be developed as an adjunctive screening tool in the noninvasive evaluation of oral lesions in vivo. This new diagnostic system may reduce the number of unnecessary biopsies.

Investigating viability of intestine using spectroscopy: a pilot study.

BACKGROUND: The differentiation of "viable" from "nonviable" bowel remains a challenge in the treatment of acute mesenteric ischemia. In this study, diffuse reflectance spectroscopy (DRS) was used to investigate the viability of bowel tissue after ischemia and reperfusion in an animal model in vivo and in real time. METHODS: A total of 25 females Sprague-Dawley rats were divided into five groups based on different bowel ischemia times. In each study group for four of them, the superior mesenteric artery was occluded using a vascular clamp for a different period (i.e., 30, 45, 60, and 90 min; n = 5 for each group). Intestinal reperfusion was accomplished by releasing the clamps after the given occlusion period for each group. Spectra were acquired by gently touching the optical fiber probe to the bowel tissue before the induce ischemia, at the end of the induced ischemia, and after the reperfusion. The data acquired before the ischemia were used as a control group. Without occluding the superior mesenteric artery, the spectra were acquired on the bowel with the same time intervals of the experiments were used as a sham group (n = 5). Subsequently, the same bowel segments were sent for histopathologic examination. RESULTS: Based on the correlation between the spectra acquired from the bowel segments and the results from the histopathologic investigation, DRS is able to differentiate the histopathologic grading that appears when the Chiu/Park score ≥5 (i.e., high-level ischemic injury) than Chiu/Park score <5. Eight out of nine low-level ischemic injury tissue samples were correctly defined using the spectroscopic classification system. All eleven high-level ischemic injury tissues that were histopathologically assigned grade 5 and above were correctly defined using the spectroscopic classification system in the ischemia-reperfusion groups. CONCLUSIONS: DRS could potentially be used intraoperatively for the assessment of bowel viability in real time. These preliminary findings suggest that DRS has the potential to reduce unnecessary resection of viable tissue or insufficient resection of nonviable tissues may reduce the mortality and morbidity rates of intestinal ischemia-reperfusion as acute mesenteric ischemia.

Elevated level of neuroserpin is an indication for the resistance to gambogic acid-induced apoptosis and oxidative stress in triple-negative breast cancer cells.

Background: The triple-negative breast cancer (TNBC) subtype, characterized by loss of HER2, estrogen, and progesterone receptors, displays aggressive phenotype and poor prognosis compared to other BC subtypes. Since the TNBC cells are devoid of receptors, endocrine therapy is an ineffective option for TNBC patients, necessitating canonical chemotherapy strategies to treat TNBC. It is crucial to use alternative and natural agents to support chemotherapy in TNBC. Objectives: To clarify the molecular mechanism of the tumorigenic effects of gambogic acid (GA) on TNBC cells with different epithelial character since GA has a wide spectrum of anticancer activity for most cancer types. Methods: We determined the cytotoxic dose of GA incubation of TNBC cells (MDA-MB-231 and BT-20 cells) for 24 h. We performed the MTT test and toluidine blue (TB) staining protocol for TNBC cells. We analyzed E-cadherin, N-cadherin, Bax, and neuroserpin mRNAs in both cells by qPCR. We evaluated apoptosis using DAPI staining and assessed the ROS using the 2',7'-dichlorofluorescin diacetate (DCFH-DA) method. Results: We determined the IC50 concentrations of GA in MDA-MB-231 and BT-20 cells to be 315.8 nM and 441.8 nM, respectively. TB staining showed that BT-20 cells survive at excessive cytotoxic doses of GA, while most of the MDA-MB-231 cells were killed. Also, we found that BT-20 cells are more resistant to GA-induced apoptosis and oxidative stress than the MDA-MB-231 cells. qPCR results showed that GA upregulated neuroserpin, an oxidative stress-relieving factor in the BT-20 cells, but not in the MDA-MB-231 cells. Conclusions: The elevated level of neuroserpin could be a predictive marker to determine the development of resistance to chemotherapeutic agents.

Diagnosis of Testicular Torsion and Differentiation From Other Pathologies Using Near-Infrared Spectroscopy.

OBJECTIVE: To develop a near-infrared (NIR) spectroscopy device to diagnose testicular torsion with high sensitivity and specificity. Specifically, we aim to investigate the differentiation between testicular torsion from other pathologies such as orchidoepididymitis, varicocele, and hydrocele. METHODS: Two LEDs with wavelengths of 660 nm and 940 nm were used as light sources in the device. Each wavelength was sent to the testicle successively, and a photodiode detected back-reflected diffuse light. The ratio of the light intensities of 660 nm and 940 nm was used as a diagnostic parameter. A multi-center clinical trial was performed in 5 different hospitals. RESULTS: In total, 62 patients in urology clinics with acute testicular pain have been recruited for the study. The developed NIR spectroscopy correctly defined all 8 testicular torsion cases. Besides, 3 orchidoepididymitis, 1 varicocele, and 3 hydrocele cases were correctly distinguished from testicular torsion. Only 1 hydrocele case was misdiagnosed as torsion. The range of the ratio was between 0.14 and 1.16 overall measurements. The ratio varied between 0.14 and 0.3 for the testicle with torsion. The ratio was between 0.49 and 1.16 for the normal testicle and testicle with other pathologies mentioned above. CONCLUSION: We have chosen the threshold ratio of 0.4 to differentiate between the normal and torsion testis and diagnosed all the torsion cases among all normal and other pathologies. The developed optical device to diagnose testicular torsion is inexpensive, user-friendly, and works based on objective criteria with high sensitivity and specificity in real time.

Investigating spectroscopic measurement of sublingual veins and tissue to estimate central venous oxygen saturation.

BACKGROUND: Venous oxygen saturation reflects venous oxygenation status and can be used to assess treatment and prognosis in critically ill patients. A novel method that can measure central venous oxygen saturation (ScvO2) non-invasively may be beneficial and has the potential to change the management routine of critically ill patients. OBJECTIVE: The study aims to evaluate the potential of sublingual venous oxygen saturation (SsvO2) to be used in the estimation of ScvO2. METHODS: We have developed two different approaches to calculate SsvO2. In the first one, near-infrared spectroscopy (NIRS) measurements were performed directly on the sublingual veins. In the second approach, NIRS spectra were acquired from the sublingual tissue apart from the sublingual veins, and arterial oxygen saturation was measured using a pulse oximeter on the fingertip. RESULTS: Twenty-six healthy subjects were included in the study. In the first and second approaches, average SsvO2 values were 75.0% ± 1.8 and 75.8% ± 2.1, respectively. The results of the two different approaches were close to each other and similar to ScvO2 of healthy persons (> 70%). CONCLUSION: Oxygen saturation of sublingual veins has the potential to be used in intensive care units, non-invasively and in real-time, to estimate ScvO2.

Early detection onset of flap failure using near infrared spectroscopy.

BACKGROUND: Near-infrared spectroscopy (NIRS) is widely used to assess flap perfusions by measuring tissue oxygen saturation (StO2). However, the StO2 level for the onset of perfusion failure is still a controversial issue. AIM: This study proposes a new threshold of StO2 level for detecting the onset of perfusion failure as early as possible to increase flap salvage rates. METHODS: Twenty patients undergoing flap surgery were included in this study - 13 flaps were implemented to cover defects that occurred due to trauma and 7 flaps to hide imperfections that occurred after cancer treatment. Thirteen flaps were in the lower extremity, six in the mandible, and one in the breast. NIRS was used to measure StO2 in 240 flap regions of the 20 patients to determine flap viability using descriptive statistics. RESULTS: The mean StO2 values from healthy flap and control regions were obtained as 81.6% ± 0.36 and 82% ± 0.18, respectively. The lowest StO2 value of 77.2% was defined as the onset of a vascular complication at a probability of 99.74% by subtracting three times the standard deviation from the mean StO2 of healthy flaps. Vascular complications were observed from 21 regions in the four flaps with StO2 values lower than 77.2%, but only one was lost. CONCLUSION: The threshold value for the onset of perfusion failure was a 5% decrease from the expected value, much lower than previously described thresholds that may facilitate the detection of perfusion failure in the early stage and increase salvage rates in flap revisions.

A new combination strategy to enhance apoptosis in cancer cells by using nanoparticles as biocompatible drug delivery carriers.

Some experimental and clinical studies have been conducted for the usage of chemotherapeutic drugs encapsulated into nanoparticles (NPs). However, no study has been conducted so far on the co-encapsulation of doxorubicin (Dox) and epoxomicin (Epo) into NPs as biocompatible drug delivery carriers. Therefore, we investigated if co-encapsulation of doxorubicin (Dox) and/or epoxomicin (Epo) into NPs enhance their anticancer efficiency and prevent drug resistance and toxicity to normal cells. We synthesized Dox and/or Epo loaded poly (lactic-co-glycolic acid) (PLGA) NPs using a multiple emulsion solvent evaporation technique and characterized them in terms of their particle size and stability, surface, molecular, thermal, encapsulation efficiency and in vitro release properties. We studied the effects of drug encapsulated NPs on cellular accumulation, intracellular drug levels, oxidative stress status, cellular viability, drug resistance, 20S proteasome activity, cytosolic Nuclear Factor Kappa B (NF-κB-p65), and apoptosis in breast cancer and normal cells. Our results proved that the nanoparticles we synthesized were thermally stable possessing higher encapsulation efficiency and particle stability. Thermal, morphological and molecular analyses demonstrated the presence of Dox and/or Epo within NPs, indicating that they were successfully loaded. Cell line assays proved that Dox and Epo loaded NPs were less cytotoxic to single-layer normal HUVECs than free Dox and Epo, suggesting that the NPs would be biocompatible drug delivery carriers. The apoptotic index of free Dox and Epo increased 50% through their encapsulation into NPs, proving combination strategy to enhance apoptosis in breast cancer cells. Our results demonstrated that the co-encapsulation of Dox and Epo within NPs would be a promising treatment strategy to overcome multidrug resistance and toxicity to normal tissues that can be studied in further in vivo and clinical studies in breast cancer.

Intraoperative assessment of laryngeal malignancy using elastic light single-scattering spectroscopy: A pilot study.

OBJECTIVES/HYPOTHESIS: The elastic light single-scattering spectroscopy (ELSSS) system is a new tool for the real-time diagnosis of cancerous lesions. In the current study, we have employed ELSSS to investigate its ability in differentiation between normal and cancerous larynx tissues ex vivo. STUDY DESIGN: Basic science study in assessment of laryngeal malignancy using spectroscopy. METHODS: ELSSS spectra of the larynx tissue were acquired using a single-fiber optical probe. Ex vivo spectroscopic measurements were acquired on 95 laryngeal lesions of 40 patients. Average slopes of the spectra in the wavelength range of 450 to 750 nm were calculated. The signs of the spectral slopes were positive for benign and negative for cancerous larynx tissues. Histopathology results were used as a gold standard to define sensitivity and specificity. RESULTS: The ELSSS system correctly defined 38 out of 41 malignant tissues as cancerous; three of them were misclassified as benign. All benign tissues were correctly classified. Moderate, severely dysplastic, and malignant tissues were correctly classified as cancerous. The system could not classify mild dysplastic tissues either benign or cancerous, whereas nearly half of them were classified as benign and the other half as malignant. The signs of the spectral slopes were used as a discrimination parameter between benign and cancerous (moderate, severely dysplastic, and malignant) lesions with a sensitivity and specificity of 94% and 100%, respectively. CONCLUSIONS: The ELSSS system has the potential to be used as an adjunctive tool in the diagnosis of cancerous laryngeal tissues in real time and noninvasively. This new diagnostic technique may reduce the number of negative biopsies. LEVEL OF EVIDENCE: NA Laryngoscope, 127:611-615, 2017.

Differentiating cancerous tissues from noncancerous tissues using single-fiber reflectance spectroscopy with different fiber diameters.

Elastic light-scattering spectra acquired with single-fiber optical probes with diameters of 100, 200, 400, 600, 800, 1000, 1200, and 1500 µm were used to differentiate cancerous from noncancerous prostate tissues. The spectra were acquired ex vivo on 24 excised prostate tissue samples collected from four patients. For each probe, the spectra and histopathology results were compared in order to investigate the correlation between the core diameters of the single-fiber optical probe and successful differentiation between cancerous and noncancerous prostate tissues. The spectra acquired using probes with a fiber core diameter of 400 µm or smaller successfully differentiated cancerous from noncancerous prostate tissues. Next, the spectra were acquired from monosized polystyrene microspheres with a diameter of 5.00±0.01 µm to investigate the correlation between the core diameters of the probes and the Mie oscillations on the spectra. Monte Carlo simulations of the light distribution of the tissue phantoms were run to interrogate whether the light detected by the probes with different fiber core diameters was in the ballistic or diffusive regime. If the single-fiber optical probes detect light in the ballistic regime, the spectra can be used to differentiate between cancerous and noncancerous tissues.

Diagnosis of testicular torsion by measuring attenuation of dual wavelengths in transmission geometry across the testis: an experimental study in a rat model.

OBJECTIVE: To develop a noninvasive and real-time dual-wavelengths optic system to detect testicular torsion in an animal model. Diagnosis of testicular torsion is challenging in emergency conditions and frequently ends with surgical exploration. MATERIALS AND METHODS: In this study, 9 male 8-month-old Wistar albino rats weighing an average of 400 g were used. A transverse incision on the upper scrotum was done and the right testis was torsed with a 720° medial rotation. The other testis of each rat was used as the control or sham group. In the sham group, the testicle was moved out of the body but no torsion was applied before repositioning into the scrotum. Transmission of continuous-wave light through all testes at wavelengths of 660 nm and 940 nm were measured. RESULTS: The ratio of the average intensities of the transmitted light of both wavelengths--660 nm to 940 nm--was used as a parameter to diagnose testis torsion. The ratios were significantly different (P = .001) between the torsion group and control group. CONCLUSION: Our noninvasive technique measuring attenuation of dual wavelengths in transmission geometry across the testis has the ability to distinguish between the testis with and without torsion on the basis of a threshold value of the ratio.