A New Method for Sperm Detection in Human Semen: Combination of Hypothesis Testing and Local Mapping of Wavelet Sub-Bands

Transcription

1 Iranian Journal of Medical Physics Vol. 9, No. 4, Auumn 2012, Received: July 29, 2012; Acceped: December 25, 2012 Original Aricle A New Mehod for Sperm Deecion in Human Semen: Combinaion of Hypohesis Tesing and Local Mapping of Wavele Sub-Bands Seyed Vahab Shojaedini 1*, Ali Kermani 2, Vahid Reza Nafisi 1 Absrac Inroducion Auomaed mehods for sperm characerizaion in microscopic videos have some limiaions such as: low conras of he video frames and possibiliy of neighboring sperms o ouch each oher. In his paper a new mehod is inroduced for deecion of sperms in microscopic videos. Maerials and Mehods In his work, firs microscopic videos are capured from specimens of human semen. Several frames of hese videos are ransformed o wavele sub-bands and bi-relaed planes are consruced from wavele sub-bands separaely. Finally, he acquired bi planes are mapped by differen local mapping funcions and decision is made using coninuiy and disconinuiy of he mapping resuls. Based on he above decision procedure, each region of he microscopic image is assigned o eiher a sperm or oher pars of semen. Resuls Performance of he proposed mehod was evaluaed by wo ses of microscopic videos which have been capured from semen of some inferile men. The firs ses belonged o semen specimens wih low densiies of sperms and he second se belonged o semen specimens wih high densiies of sperms. Conclusion The resuls of his sudy revealed ha he proposed mehod in his work is more efficien in sperm deecion and exracion compared wih he curren approaches in boh scenarios. Furhermore, i is eviden ha for specimens wih higher sperm densiies he proposed mehod improved sperm deecion also reduces false deecion rae considerably. Keywords: Deecion, Hypohesis Tesing, Mapping, Microscopic Video, Semen, Wavele Sub-Bands 1- Elecrical Engineering Deparmen, Iranian Research Organizaion for Science and Technology, Tehran, Iran *Corresponding auhor: Tel: ; shojadini@iros.org 2- Elecrical Engineering Deparmen, Iran Universiy of Science and Technology Iran J Med Phys, Vol. 9, No. 4, Auumn

2 Seyed Vahab Shojaedini e al. 1. Inroducion Inferiliy is a medical condiion faced by 15% of couples worldwide [1]. Reliable repors have deermined ha male impoence is he main facor in 30-40% of all he inferiliy cases [2]. Men feriliy depends on several facors, bu he sperms have he key role in his procedure. Therefore, esimaing sperm parameers has been one of he mos favorie policies of researchers in inferiliy monioring and cure [3, 4]. In recen years, modern microscopic imaging has provided abiliy of analyzing of semen [5-8]. In his mehod, he poenial of feriliy in men is deermined by compuing he parameers of sperms exising in microscopic images of semen. Some of hese parameers are size, morphology, and heir moion rajecory [9]. Based on above separaing sperms from oher paricles of semen is a vial sep for analyzing feriliy [10]. Several mehods have been used for such a discriminaion which he oldes one is manually sperm deecion by an exper person. Unforunaely his mehod is ime-consuming and is performance is degraded by human errors [11]. Therefore, he auomaed mehods have been subsiued for processing hese images. The main challenges which limi he performance of he auomaed mehods are: he low conras of he video frames, changing number of acive sperms in several frames, and finally he possibiliy of neighboring sperms o ouch each oher so hey may be considered as merged sperms [12]. Several approaches have been proposed o solve problems of auomaic sperm deecion mehods. Some approaches uilize pixel-based echniques for separaing sperms from oher semen paricles. For insance in [13] sperms have been deeced using a hreshold which has been calculaed direcly from inensiy of pixels of microscopic images. In some oher improved echniques, he above hreshold changes adapively during several successive frames [14]. In some researches, several ypes of 2D wavele ransform have been uilized for disinguishing of sperms from oher pars of he semen [15-17]. Alhough such pixel-based mehods have improved speed in sperm deecion bu hey ofen cause considerable errors in low SNR microscopic images because of heir sensiiviy o noise [18]. There is a class of mehods ha separaes sperms by using conneciviy of pixels. Differen conour-based mehods may be suiable examples for he above class. Despie such mehods show grea abiliy o exrac curve-shape boundaries belonging o head of sperms, hey ofen need complemenary pos-processing o exrac ail of sperms [19]. In anoher class of mehods, he disconinuiy beween sperms and oher paricles of semen (e.g., background of microscopic image) is used o discriminae hem. Several mehods which are based on edge, gradien, Hilber ransform, and waershed segmenaion are examples of he mehods of his class which mainly handle he problem of exracing false paricles in semen [20, 21]. In his paper, a new mehod is inroduced for separaing sperms from oher paricles of semen in microscopic video which works based on hypohesis esing. In he proposed mehod, firs he 2D wavele ransform is applied on he microscopic image. In he second sep, bi-plane images are consruced using he acquired wavele sub-bands. Then, bi-plane images are mapped o conneciviy space which leads o four mapped images. Finally, dependence of each pixel o sperm or he background is deermined for mapped images using is conneciviy facors. The biplane image processing is differen for several images which are resuled from wavele subbands. Thus, i may be concluded ha he decision funcion of hypohesis esing for each region of microscopic image is consruced based on local characerisics of hose regions. Therefore, such a decision funcion leads o more accurae decision abou he presence of sperms in each region. In secion 2, he proposed algorihm is described mahemaically which includes acquisiion obaining wavele sub-bands, biplanes, mapping, and consrucion decision funcion for hypohesis esing. In secion 3, he performance of he proposed mehod is 284 Iran J Med Phys, Vol. 9, No. 4, Auumn 2012

3 Hypohesis Tesing Based Sperm Deecion evaluaed for wo real scenarios, one wih low densiy of sperms and he oher one wih high densiy of sperms. In secion 4, he experimenal resuls are compared wih resuls of several exising mehods using heir acual parameers. Conclusion is presened in he las secion of he manuscrip. 2. Maerials and Mehods Suppose I as a microscopic video capured I from semen specimen and is one of is frames which is appeared in ime slo. The image I includes sperms, plasma, and debris. In his aricle he wo laer parameers are called arifac. Also, i conains some amoun of noise. For each pixel of I i can be wrien: I I ( m, j) 1 m M, 1 j J, 1 T (1) In equaion (1), I is he inensiy of a pixel in I which is locaed in row and column equal wih m and j, respecively. Also, M and J are he image sizes and T is he ime lengh of video I. A differen image is formed by he wo frames of I which appeared in ime slos and 0 as: D I I ) mj (2) ( 0 In which D is he inensiy of he pixel which is locaed in row and column equal wih m and j of he differen image D, respecively. Now, dependence of D sperm or arifac is deermined by using hypohesis esing which is inroduced in equaion (3). H 0 : H1 : D D g s n g n (3) In his equaion H0 shows he dependence of D o arifac and noise while H1 shows is o dependence o sperm. Also s, g and n show sperm, arifac and noise componens in D. Compuing he second level discree wavele ransform [22] of obained which are called elemens are C D four sub-bands is C which heir ha are locaed in row and column equal wih m and j, respecively. These four sub-bands are menioned as: {, HL, LH, } (4) The above se may be decomposed o sub-ses ' and " as: " ' { }, { HL, LH, } (5) HL In he nex sep, each of he sub-bands C, LH C andc bi-planes such as are convered o K number of ' Pk as: P ' ' k P k ( m, j), k 1,2,..., K (6) In equaion (6), he erm ' Pk shows he bi which is locaed in posiion ( m, j) of bi-plane k which is obained from C '. The maximum number of bi-planes is obained from equaion (7) [23]: K 2 max( ), ( m, j) C (7) Now, he procedure which is menioned bellow is applied o each bi of he bi-planes The feaure se C C is exraced from a sliding window which is cenered in ( m, j) and is size is m 1) ( j 1) as: ( 0 0 C C C C ( mm0 )( j j ),...,,..., 0 ( mm0 )( j j0 ) 2.2. For C he mapping parameer U (8) defined based on changes of he values of above sliding window as: is Iran J Med Phys, Vol. 9, No. 4, Auumn

4 Seyed Vahab Shojaedini e al. U u 0 (9) 2.3. The mapping funcion is defined for all elemens of C based on mapping parameer as: U E C In which U E ( C C ) shows mapped pixels of (10) C which make a mapped pseudo-image E. Also, C definiion of shows he mean ofc equaion (10) shows ha. Based on U (e.g., equaion 9), he E on coninuiy and dis-coninuiy of C E srongly depends. Now, is convered o K number of bi-planes P' k in such way ha equaion (11) can be wrien similar o equaion (6) as: P' k P' k ( m, j) : k 1,2,..., K (11) 2.4. Consider LH Pk and is wo horizonal neighbors. If a leas wo of hese hree bis are LH equal o 1 hen P' 1 is subsiued, k k LH oherwise P' 0 is replaced Consider HL Pk and is wo verical neighbors. If a leas wo of hese hree bis are HL equal o 1 hen P' 1 is subsiued, k k HL oherwise P' 0 is replaced Consider If u differen values exis in window If here are no differen values Pk and is four diagonal neighbors. If a leas hree of hese five bis are equal o 1 hen P' 1 is subsiued, k k oherwise P' 0 is replaced. By running above seps, he four bi-planes LH HL P', P', P', and P' are obained in accordance wih four sub-bands of. Now, he processed image F is obained by applying inverse 2D wavele ransform on wavele sub-bands are P', P', P' P'. Finally, sperms in F LH P' which is HL and are exraced as conneced objecs which are lised in equaion (12), by using he labeling mehod which has been inroduced in [24]. O O O,..., O 1,..., w W (12) In which O is he se of deeced sperms in ime slo, Ow is he sperm w of his se and W is he oal number of exraced sperms. Combining equaions (3) and (13) dependence of each pixel of microscopic image is deermined o sperm or arifac as: H 0 : D H1 : D F F s g O n O g n (13) Two sandard parameers are defined o evaluae he performance of he proposed mehod. The firs parameer is probabiliy of deecion and shows he abiliy of mehod in exracing correc sperms in presence of arifac and noise. I is defined as: P D P F O H ) (14) ( 1 The second parameer is false deecion error (false alarm) which shows he probabiliy of deecing oher semen paricles as sperms. This parameer is defined as: P fa P F O H ) (15) 3. Resuls ( 0 The proposed algorihm was applied o microscopic videos which were capured from human semen. The daa collecion seup conained a Nikon inver microscope using a 40x zoom lens and a Moicom480 digial CCD camera which was mouned on he microscope o record video. This seup is showed in figure 286 Iran J Med Phys, Vol. 9, No. 4, Auumn 2012

5 Hypohesis Tesing Based Sperm Deecion (1-a). A calibraed microscope slide was used for all experimens. This microscope slide is showed in figure (1-b) and i was scaled per 10 micromeer which enabled us o esimae size and movemen parameers of sperms. implemened using Malab Furhermore, Hisogram Thresholding Segmenaion algorihm (HTS) [25], Muli Srucure Elemen Segmenaion (MSES) [26], and Dynamic Threshold Segmenaion (DTS) [14] were implemened o compare he resuls wih he proposed algorihm. The capured videos firsly were processed manually o obain a reference deecion o compare wih he four menioned mehods. Then, he performance of each algorihm was deermined by compuing is PD and Pfa as described in equaions (14) and (15). (a) Figure 2. Reference image from empy microscope slide (b) Figure 1. Daa collecion seup: (a) inver microscope, zoom lens and digial CCD, (b) calibraed and scaled microscope slide. Afer calibraion, he semen sample was pu on a non-scaled microscope slide and digial videos were capured. The image of his nonscaled microscope slide exised in all capured frames as background. Therefore, he image of empy non-scaled microscope slide (which has been showed in figure (2) was subraced from all capured frames for background eliminaion. Using his procedure, 135 frames of semen were invesigaed which belonged o 9 inferile men. The proposed mehod was Tess were carried ou on wo differen scenarios which in one of hem, he semen specimens had low densiies of sperms (bellow 2 million sperms per millilier) so hey were beer disinguished in capured images. In he second scenario, specimens had high densiies of sperms (more han 2 million sperms per millilier), so heir near disances led o degrading of deecion. Specificaions of boh scenarios are been showed in able 1. Iran J Med Phys, Vol. 9, No. 4, Auumn

6 Seyed Vahab Shojaedini e al. Specificaion Min and Max sizes of sperms Frame size (pixels) Number of samples Frame rae Table1. Specificaions of es scenarios Value Specificaion 5, 50 pixels 288*352 9 persons 30 frames per second Number of capured frames Min-Max velociy of sperms Average conras Average noise change o sperm changes Value pixels per frame 15% 0.75 Figure (3) shows resuls produced by he firs scenario. Figure (3-a) shows he capured frame in which he sperms labeled manually while (3-b) and (3-c) show he resuls of processing his frame using he proposed mehod and HTS. I is obvious ha he proposed mehod has exraced all 8 sperms which were labeled in figure (3-a) while HTS exraced only 5 sperms which one of hem was exraced incompleely. Also, i can be seen ha boh mehods have no exraced any false sperm. Figure (4) shows one of he obained resuls on capured images belonging o he second scenario. Figure (4-a) shows he capured frame in which he sperms labeled manually while (4-b) and (4-c) show he resuls of processing of his frame uilizing he proposed and HTS mehods. I may be considered ha he proposed mehod exraced 25 sperms (one of hese sperms has been exraced incompleely) from oal 32 manually labeled sperms while HTS exraced 12 sperms which four of hem have been exraced incompleely. Also, i may be noed ha alhough he proposed mehod exraced only one false sperm, he HTS deeced 24 false sperms. 4. Discussion Daa which were obained from microscopy of semen specimens wih low and high densiies of sperms were invesigaed. The proposed algorihm and hree exising mehods (MSES, DTS, and HTS) were implemened and he resuls were compared wih manual deecion. Probabiliies for rue and false deecion were used as parameers for comparison. To compue probabiliy of deecion, wo definiions for rue deecion were applied. In some inferiliy diagnosic applicaions, he only imporan parameer is he sperm populaion [27], while for oher applicaions, he complee exracion of sperms is necessary because of he necessiy o compue heir lengh, morphology, and size. Based on hese wo definiions, probabiliy of deecion was compued in wo differen ways. In he firs manner, he rue exraced sperm was considered as a sperm wih correc posiion in he image. In he second manner, he correc deecion was considered as a sperm ha a leas 90% of is pixels were exraced correcly. Table 2 shows differen performances for he proposed mehod and oher sperm exracion echniques. Based on his able, in he firs scenario, he resuls of he proposed mehod were 43%, 39%, and 37% beer han DTS, MSES, and HTS by using probabiliy of deecion wihou size crieria as he comparison parameer. Theses superioriies reached 46%, 44%, and 40% when probabiliy of deecion wih size crieria was used as he comparison parameer. Comparing false deecion probabiliies of hese four mehods shows ha despie of considerable superioriy in deecion by he proposed algorihm, is false deecion probabiliy did no increase compared wih oher echniques. Table 2 shows more considerable superioriy for he proposed mehod in he second scenario. In his scenario, he resuls which have been obained by using he proposed mehod were 43%, 41%, and 38% beer han DTS, MSES, and HTS when he probabiliy of deecion wihou size crieria was used as he comparison parameer. 288 Iran J Med Phys, Vol. 9, No. 4, Auumn 2012

7 Hypohesis Tesing Based Sperm Deecion (a) (b) (c) Figure 3. One of he resuls from he firs scenario: (a) The capured microscopic images for a semen specimen in which he sperms have been labeled manually, (b) Exraced sperms using he proposed algorihm, and (c) Exraced sperms using HTS algorihm. (a) (b) (c) Figure 4. One of he resuls from he second scenario: (a) The capured microscopic images for a semen specimen in which he sperms have been labeled manually, (b) Exraced sperms using he proposed algorihm, and (c) Exraced sperms using HTS algorihm. Iran J Med Phys, Vol. 9, No. 4, Auumn

8 Seyed Vahab Shojaedini e al. Table 2. Comparing he performances of he implemened algorihms in boh of scenarios Comparison Parameer Probabiliy of Deecion (wihou size crieria) Probabiliy of Deecion (wih size crieria) Probabiliy of False Deecion Images capured from semen wih Low Densiy of Sperms (Firs Scenario) Proposed DTS MSES HTS Mehod Images capured from semen wih High Densiy of Sperms (Second Scenario) Proposed DTS MSES HTS Mehod By using probabiliy of deecion wih size crieria as he comparison parameer, he superioriies of he proposed mehod agains DTS, MSES, and HTS reached 53%, 52%, and 48%, respecively. I mus be noed ha he deecion probabiliies of DTS, MSES, and HTS in he second scenario were obained in parallel wih increasing heir false deecion probabiliy up o 51%, 47%, and 48% while false deecion probabiliy for he proposed mehod in his scenario was 5% which doesn show he meaningful increase due o is value in he firs scenario. 5. Conclusion In his manuscrip, a new mehod is inroduced for separaing sperms from oher semen paricles in microscopic videos. In he proposed mehod, firsly a hypohesis esing funcion is defined o deermine he dependence of each pixel o sperm or oher paricles. Then, each hypohesis is evaluaed using a decision funcion which is consruced by mapping he wavele sub-bands of each video frame o several bi-planes. In order o evaluae he performance of he proposed mehod, wo scenarios are carried ou based on microscopic videos of semen specimens conaining differen densiies of sperms. The firs scenario belonged o semen specimens wih low densiies of sperms and he second one deal wih specimens wih high densiies of sperms. In boh scenarios, he performance of he proposed algorihm is compared wih some exising mehods (DTS, MSES, and HTS) by using hree sandard parameers. The firs parameer is probabiliy of deecion when he deeced sperm is defined as sperm wih correc posiion in frame. The second parameer is probabiliy of deecion when he deeced sperm is considered as a sperm and a leas 90% of is pixels are deeced correcly. The hird parameer is probabiliy of false deecion. The resuls show he superioriy of he proposed mehod in boh of scenarios using all above menioned parameers. In he firs scenario, he proposed mehod deeced 85% of sperms wih size crieria which is a leas 40% improvemen compared wih oher exising mehods. Also, using deecion probabiliy wihou size crieria, he proposed mehod shows a leas 37% improvemen compared wih oher exising mehods. These improvemens are obained in false deecion probabiliy equal wih 4% for he proposed mehod which shows no degradaion compared wih oher mehods. Alhough in he second scenario he deecion parameers of all mehods decreases due o high densiy of sperms, he superioriy of he proposed mehod compared wih oher echniques increases in his scenario. In capured videos wih high densiies of sperms, he proposed algorihm shows a leas 48% and 38% improvemens 290 Iran J Med Phys, Vol. 9, No. 4, Auumn 2012

9 Hypohesis Tesing Based Sperm Deecion compared wih oher mehods in deecion probabiliy wih and wihou size crieria, respecively. Finally, in his scenario, he improvemen in deecion using he proposed mehod is paralleled wih improvemen in false deecion parameer in such a way ha he false deecion probabiliy of he proposed mehod is a leas 42% beer han oher echniques. Consequenly, i can be concluded ha he proposed mehod may be used as a suiable alernaive for deecing sperms in microscopic videos especially in semen specimens wih high densiies of sperms. Acknowledgemens The auhors are hankful o Dr Shahrokh Farahmand for faihful discussions and ediing pars of his aricle. References 1. Speroff L, Friz MA. Clinical gynecologic endocrinology and inferiliy: Lippinco Williams & Wilkins; Regulaed Feriliy Services: A commissioning aid, Deparmen of Healh, UK, June Available a: hp://dh.gov.uk/prod_consum_dh/groups/dh_digialasses/documens/digialasse/dh_ pdf. Accessed Jan 28, Wenzhong Y, Shuqun S. Auomaic Chromosome Couning Algorihm Based on Mahemaical Morphology. Journal of Daa Acquisiion & Processing. 2008;23(9): Menkveld R, Wong WY, Lombard CJ, Wezels AM, Thomas CM, Merkus HM, e al. Semen parameers, including WHO and sric crieria morphology, in a ferile and subferile populaion: an effor owards sandardizaion of in-vivo hresholds. Hum Reprod Jun;16(6): Leung C, Lu Z, Esfandiari N, Casper RF, Sun Y, ediors. Deecion and racking of low conras human sperm ail. Auomaion Science and Engineering (CASE), 2010 IEEE Conference on; 2010: IEEE. 6. Shi LZ, Nascimeno J, Chandsawangbhuwana C, Berns MW, Bovinick EL. Real-ime auomaed racking and rapping sysem for sperm. Microsc Res Tech Nov;69(11): Abbiramy V, Shanhi V, Allidurai C, ediors. Spermaozoa deecion, couning and racking in video sreams o deec ashenozoospermia. Signal and Image Processing (ICSIP), 2010 Inernaional Conference on; 2010: IEEE. 8. Abbiramy V, Shanhi V. Spermaozoa segmenaion and morphological parameer analysis based deecion of eraozoospermia. Inernaional Journal of Compuer Applicaions IJCA. 2010;3(7): Carrillo H, Villarreal J, Soaquira M, Goelkel M, Guierrez R, ediors. A compuer aided ool for he assessmen of human sperm morphology. Bioinformaics and Bioengineering, 2007 BIBE 2007 Proceedings of he 7h IEEE Inernaional Conference on; 2007: IEEE. 10. Carrillo H, Villarreal J, Soaquira M, Goelkel A, Guierrez R, ediors. Spermaozoon segmenaion owards an objecive analysis of human sperm morphology. Image and Signal Processing and Analysis, 2007 ISPA h Inernaional Symposium on; 2007: IEEE. 11. Acosa AA, Kruger TF.Menkveld R, Kruger T. Evaluaion of sperm morphology by ligh microscopy. Human Spermaozoa in Assised Reproducion. 1996: Mazzilli F, Rossi T, Delfino M, Nofroni I. Applicaion of he upgraded image superimposiion sysem (SIAS) o he assessmen of sperm kinemaics. Andrologia Jul;31(4): Wu Q, Merchan F, Casleman KR. Microscope image processing: Academic press; Xuan ZL, Yan WZ, ediors. The sperm video segmenaion based on dynamic hreshold. Machine Learning and Cyberneics (ICMLC), 2010 Inernaional Conference on; 2010: IEEE. 15. Cui C. Per-Pixel Background Esimaion In Video Microscopy Using Frame Grouping and Wavele Based Image Fusion Liu JC, Hwang WL, Chen MS, Tsai JW, Lin CH, ediors. Wavele based acive conour model for objec racking. Image Processing, 2001 Proceedings 2001 Inernaional Conference on; 2001: IEEE. 17. Yi W, Park K, Paick J, ediors. Parameerized characerizaion of ellipic sperm heads using Fourier represenaion and wavele ransform. Engineering in Medicine and Biology Sociey, 1998 Proceedings of he 20h Annual Inernaional Conference of he IEEE; 1998: IEEE. 18. Meijering E, Smal I, Dzyubachyk O, Olivo-Marin JC. Time-lapse imaging. Microscope Image Processing. 2008: Iran J Med Phys, Vol. 9, No. 4, Auumn

10 Seyed Vahab Shojaedini e al. 19. Zimmer C, Labruyere E, Meas-Yedid V, Guillen N, Olivo-Marin JC. Segmenaion and racking of migraing cells in videomicroscopy wih parameric acive conours: A ool for cell-based drug esing. Medical Imaging, IEEE Transacions on. 2002;21(10): Wählby C, SINTORN IM, Erlandsson F, Borgefors G, Bengsson E. Combining inensiy, edge and shape informaion for 2D and 3D segmenaion of cell nuclei in issue secions. Journal of Microscopy. 2004;215(1): Yang X, Li H, Zhou X. Nuclei segmenaion using marker-conrolled waershed, racking using mean-shif, and kalman filer in ime-lapse microscopy. Circuis and Sysems I: Regular Papers, IEEE Transacions on. 2006;53(11): Malla S. A wavele our of signal processing Academic, San Diego, CA. 23. Gonzalez RC, Woods RE. Digial Image Processing: Pearson/Prenice Hall; Haralick RM, Shapiro LG. Compuer and Robo Vision. Addison-Wesley Pub. Co. Boson, Liu B, Chen X, Ma C, Zhang D, Zhou X, He Y, ediors. Research on hreshold segmenaion in racking echnology of moving objecs. Indusrial Mecharonics and Auomaion (ICIMA), nd Inernaional Conference on; 2010: IEEE. 26. Bai W, Liu J, Chen G, ediors. Segmenaion of whie ra sperm image. Proceedings of SPIE, he Inernaional Sociey for Opical Engineering; 2011: Sociey of Phoo-Opical Insrumenaion Engineers. 27. Morimer ST. A criical review of he physiological imporance and analysis of sperm movemen in mammals. Human Reproducion Updae. 1997;3(5): Iran J Med Phys, Vol. 9, No. 4, Auumn 2012